CN101553236A - Compounds and methods for modulating expression of GCCR - Google Patents

Abstract

The disclosure describes short antisense compounds, including such compounds comprising chemically modified high affinity monomers that are 8-16 monomers in length. Certain such short antisense compounds can be used to reduce target nucleic acids and/or proteins in cells, tissues, and animals with increased potency and improved therapeutic index. Thus, the present invention provides short antisense compounds comprising high affinity nucleotide modifications useful for reducing target RNA in vivo. Such short antisense compounds are also effective at lower doses than previously described antisense compounds, which allows for reduced toxicity and therapeutic costs. In addition, the short antisense compounds described have greater potential for oral administration.

Description

Regulate the Compounds and methods for of the expression of GCCR

The application is the part continuation application of the PCT application PCT/US2007/061183 of submission on January 27th, 2007, and the priority of therefore applying for by this PCT of 35 U.S.C. §, 120 requirements.The U.S. Provisional Application series number 60/746 that the application also submits to by 35 U.S.C. § 119 (e) requirements on Mays 5th, 2006,631, the U.S. Provisional Application series number of submitting on May 11st, 2,006 60/747,059, the U.S. Provisional Application series number of submitting on June 23rd, 2,006 60/805, the priority of the U.S. Provisional Application series number 60/864,554 that on November 6th, 660 and 2006 submitted to.The full content of each of these applications and all lists of references of wherein quoting all are attached to herein for any and all purpose integral body by reference.

Sequence table

The application submits to together with the sequence table of electronic format.Sequence table is to provide as the title of the 700Kb size of creating on May 7th, 2007 file for CORE0061WO12SEQ.TXT.Information in the sequence table of electronic format integral body by reference is attached to herein.

Background of invention

This way of targeting Disease-causing gene sequence is initial (the Belikovaet al. that proposes before nearly 40 years, Tet.Lett., 1967,37,3557-3562), in cell culture, proved antisense activity (Zamecnik et al. after having spent 10 years, Proc.Natl.Acad.Sci.U.S.A., 1978,75,280-284).Antisense technology is derived from the disease of Disease-causing gene or an advantage in the disease is in treatment, and it is the direct genetic method that can regulate the expression of specific Disease-causing gene.

In general, thus the principle of antisense technology is antisense compounds and target nucleic acid, and the adjusting that hybridization realizes activity of gene expression or function (as transcribe, translation or montage) takes place.The adjusting of gene expression can realize by for example target degraded or based on the inhibition that occupies (occupancy-basedinhibition).Regulate an example of RNA target function by degraded, the degraded that is taken place when being the hybridization of target RNA and DNA sample antisense compounds based on RNA enzyme H.Another example of degrading regulator gene to express by target is that RNA disturbs (RNAi).RNAi is a kind of form of the gene silencing of antisense mediation, relates to introduce dsRNA and cause being reduced by the sequence-specific of the endogenous mRNA level of targeting.Sequence-specific makes antisense compounds extremely attractive in the following areas: determine the research tool of the instrument of (gene functionalization) and evaluation and characterisation of nucleic acids enzyme and regulate the medicine of any pathogenetic expression of gene that relates to multiple disease as selectivity as target validation (target validation) and gene function.

Antisense technology is the effective ways that reduce the expression of one or more specific gene products, and is therefore provable particularly useful in multiple treatment application, diagnostic application and research are used.Nucleoside through chemical modification is used to be incorporated in the antisense compounds usually, strengthening one or more characteristics, as nuclease resistance, pharmacokinetics or to the affinity of target RNA.

Although the expansion of relevant knowledge since the invention antisense technology is higher for effect (efficacy), toxicity is less and lower-cost antisense compounds still has the demand that is not met.Till present disclosure, high-affinity is modified (high-affinity modification) and is not applied to yet designing in order to reduce the short antisense compounds of target RNA in the body.This is because worry that it has the target specificity of much degree when the target that 15 nucleotide or shorter sequence should be used for reducing in the living systems.Research is before reported, can realize higher specificity with the antisense compounds of length 16-20 nuclear base (nucleobase), and therefore realizes bigger usefulness (potency) potentiality.

Present disclosure explanation will be incorporated in the antisense compounds through the high-affinity nucleotide of chemical modification, can make the about 8-16 of length the short antisense compounds of examining base can be used for reducing target RNA in the animal with the usefulness of raising and improved therapeutic index.Therefore, this paper provides so short antisense compounds, and it comprises the high-affinity nucleotide modification that can be used for reducing target RNA in the body.This short antisense compounds is also effective under than the lower dosage of the antisense compounds of describing before, makes toxicity and treatment cost reduce.

Summary of the invention

Disclosed herein is that short antisense compounds and the described chemical compound of use reduce the method that the target RNA in the cell or tissue is expressed.In certain embodiments, provided herein is the method that reduces the expression of the target in the animal, and described method comprises that the short antisense compounds with the nucleic acid of this target of targeting gives this animal.In some embodiment Shen, short antisense compounds is the oligonucleotide chemical compound.In certain embodiments, short antisense oligonucleotide length be about 8-16, preferably 9-15, more preferably 9-14, more preferably 10-14 nucleotide and comprise interval region (gapregion), the both sides of this interval region respectively are connected to pterion (wing), and wherein each pterion is made up of 1-3 nucleotide independently.Preferred die body (motif) includes but not limited to pterion-deoxidation interval-pterion die body of 3-10-3, the 2-10-3,2-10-2,1-10-1,2-8-2,1-8-1,3-6-3 or the 1-6-1 that are selected from.In a preferred embodiment, lack antisense oligonucleotide and comprise at least one high-affinity modification.In another embodiment, high-affinity is modified the high-affinity nucleotide that comprises through chemical modification.In a preferred embodiment, each pterion is made up of 1-3 the nucleotide of modifying through high-affinity independently.In one embodiment, the nucleotide of modifying through high-affinity is through sugar-modified nucleotide.

In certain embodiments, lacking antisense compounds, to demonstrate the uptake ratio of comparing in intestinal with the longer antisense compounds of length higher.Therefore, this paper also provides the method that reduces the target in the animal, and described method comprises orally give short antisense compounds of the present invention.

In certain embodiments, lack the proteinic nucleic acid that antisense compounds targeting coding is selected from ApoB, SGLT2, PCSK9, SOD1, CRP, GCCR, GCGR, DGAT2, PTP1B and PTEN.

Provide the method for the metabolic disease in the treatment animal in addition, described method comprises the animal that so short antisense compounds is needed this treatment, and this weak point antisense compounds targeting relates to the nucleic acid of regulating glucose metabolism or removing, lipid metabolism, cholesterol metabolism or insulin signaling transduction.

Also provide insulin sensitivity, the reduction blood-glucose that increases in the animal or reduce HbA_1cMethod, described method comprises and gives described animal with so short antisense compounds that this weak point antisense compounds targeting coding relates to the nucleic acid of the target of regulating glucose metabolism or removing, lipid metabolism, cholesterol metabolism or insulin signaling transduction.

Provide the total serum cholesterol that reduces in the animal in addition, serum LDL, serum VLDL, serum hdl, serum triglycerides, the method of serum apolipoprotein (a) or free fatty, described method comprises and gives described animal with so short antisense compounds, this weak point antisense compounds targeting coding relates to regulates glucose metabolism or removing, lipid metabolism, the nucleic acid of the target of cholesterol metabolism or insulin signaling transduction, wherein said short antisense compounds length is 8-16 nucleotide and comprises interval region, the both sides of this interval region respectively are connected to the pterion, and wherein each pterion is made up of 1-3 the nucleotide of modifying through high-affinity independently.

Relate to some target of regulating glucose metabolism or removing, lipid metabolism, cholesterol metabolism or insulin signaling transduction and include but not limited to GCGR and ApoB-100.Therefore, provide the short antisense compounds and the method that reduces described target and/or the expression of target nucleic acids in animal of the nucleic acid of targeting coding GCGR and ApoB-100.In addition, the short antisense compounds that provides the nucleic acid of targeting coding GCGR and ApoB-100 is used for the treatment of the purposes of disease metabolic or cardiovascular or disease.

In certain embodiments, lack antisense compounds and also comprise conjugated group (conjugategroup).Conjugated group includes but not limited to C₁₆And cholesterol.

In certain embodiments, lacking antisense compounds comprises and connects key (internucleoside linkage) or sugar moieties between at least one modified nuclear base, nucleoside.In certain embodiments, connecting key between this modified nucleoside is to connect key between thiophosphate (phosphorothioate) nucleoside.In certain embodiments, connecting key between each modified nucleoside is to connect key between the thiophosphate nucleoside.

In certain embodiments, lack antisense compounds and comprise at least one high-affinity modification.In some this embodiment, the high-affinity modification is the high-affinity nucleotide through chemical modification.In certain embodiments, the high-affinity nucleotide through chemical modification is through sugar-modified nucleotide.In certain embodiments, in sugar-modified nucleotide, have at least one between 4 ' and 2 ' position of sugar, to comprise abutment (bridge).Each independently is β-D or α-L sugar conformation through sugar-modified nucleotide.In certain embodiments, each described nucleotide of modifying through high-affinity is given every nucleotide Δ T of 1-4 degree (degree) at least_mIn certain embodiments, each describedly comprises 2 '-substituent group outside H or the OH through sugar-modified nucleotide.Thisly comprise that through sugar-modified nucleotide those have the nucleotide of the dicyclo sugar moieties of 4 '-2 ' bridge joint.In certain embodiments, each 2 '-substituent group independently is alkoxyl, substituted alkoxy or halogen.In certain embodiments, each 2 '-substituent group is OCH₂CH₂OCH₃(2 '-MOE).

In certain embodiments, short antisense compounds have one or more between 4 ' and 2 ' position of sugar, comprise abutment through sugar-modified nucleotide, wherein each described abutment independently comprises 2-4 and independently is selected from-[C (R₁) (R₂)]_n-,-C (R₁)=C (R₂)-,-C (R₁)=N-,-C (=NR₁)-,-C (=O)-,-C (=S)-,-O-,-Si (R₁)₂-,-S (=O)_x-and-N (R₁)-linking group;

Wherein

X is 0,1 or 2;

N is 1,2,3 or 4;

Each R₁And R₂Independent is H, protecting group, hydroxyl, C₁-C₁₂Alkyl, substituted C₁-C₁₂Alkyl, C₂-C₁₂Thiazolinyl, substituted C₂-C₁₂Thiazolinyl, C₂-C₁₂Alkynyl, substituted C₂-C₁₂Alkynyl, C₅-C₂₀Aryl, substituted C₅-C₂₀Aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C₅-C₇Alcyl, substituted C₅-C₇Alcyl, halogen, OJ₁, NJ₁J₂, SJ₁, N₃, COOJ₁, acyl group (C (=O)-H), substituted acyl group, CN, sulfonyl (S (=O)₂-J₁) or sulfinyl (S (=O)-J₁); With

Each J₁And J₂Independent is H, C₁-C₁₂Alkyl, substituted C₁-C₁₂Alkyl, C₂-C₁₂Thiazolinyl, substituted C₂-C₁₂Thiazolinyl, C₂-C₁₂Alkynyl, substituted C₂-C₁₂Alkynyl, C₅-C₂₀Aryl, substituted C₅-C₂₀Aryl, acyl group (C (=O)-H), substituted acyl group, heterocyclic radical, substituted heterocyclic radical, C₁-C₁₂Aminoalkyl, substituted C₁-C₁₂Aminoalkyl or protecting group.

In one aspect, each described abutment independently is-[C (R₁) (R₂)]_n-,-[C (R₁) (R₂)]_n-O-,-C (R₁R₂)-N (R₁)-O-or-C (R₁R₂)-O-N (R₁)-.In yet another aspect, each described abutment independently is 4 '-(CH₂)₃-2 ', 4 '-(CH₂)₂-2 ', 4 '-CH₂-O-2 ', 4 '-(CH₂)₂-O-2 ', 4 '-CH₂-O-N (R₁)-2 ' and 4 '-CH₂-N (R₁)-O-2 '-, each R wherein₁Independent is H, protecting group or C₁-C₁₂Alkyl.

In certain embodiments, this paper provides and can be used for reducing the target relevant with the morbid state in the animal and/or the short antisense compounds of target RNA.In certain embodiments, provide short antisense compounds in order to the expression that reduces the target RNA in the animal.In certain embodiments, this paper provides short antisense compounds preparing in order to the purposes in the medicine of the metabolic disease in the treatment animal.In certain embodiments, this paper provides short antisense compounds preparing in order to increase insulin sensitivity, the reduction blood-glucose in the animal or to reduce HbA_1cMedicine in purposes.Also providing short antisense compounds is preparing in order to the purposes in the medicine that reduces total serum cholesterol, serum LDL, serum VLDL, serum hdl, serum triglycerides, serum apolipoprotein (a) or free fatty in the animal.

In certain embodiments, short antisense compounds provided herein is compared with the longer parent antisense oligonucleotide of 20 nucleotide of length at least, strikes in target RNA to subtract to show the usefulness that equates or improve aspect (knockdown).In certain embodiments, lacking antisense compounds compares with the parent antisense oligonucleotide and demonstrates onset faster (target RNA minimizing).In certain embodiments, the usefulness of raising is in kidney.In certain embodiments, target RNA is mainly expressed in kidney.In certain embodiments, the usefulness of raising is in liver.In certain embodiments, target RNA is mainly expressed in kidney.

Detailed Description Of The Invention

It should be understood that above summary of the invention and detailed Description Of The Invention hereinafter all only are exemplary and indicative, do not limit the present invention of institute's prescription.The singular noun of using in this article comprises plural implication, unless clearly indicate really not so." or (perhaps) " that this paper uses mean " and/or ", unless otherwise prescribed.In addition, used term " comprises ", " comprising " be nonrestrictive.Equally, key element and the component that comprises a unit (unit) both contained in term such as " key element (element) " or " component (component) ", also contain and comprise key element and the component that surpasses a subunit (subunit), unless clearly indicate really not so.

Chapter title used herein only for the purpose of style of writing tissue, can not be interpreted as limiting described theme.The All Files that refers among the application or the each several part of file include but not limited to patent, patent application, article, books and paper, all for any purpose by reference clearly integral body be attached to herein.U.S. Patent Application Serial 10/712,795 and 10/200,710 for any purpose by reference clearly integral body be attached to herein.

A. definition

Unless provide specific definition, otherwise used analytical chemistry described herein, synthetic organic chemistry and medical science and pharmaceutical chemical term and these chemical method and the technology of relating to is term and method and technology known in this field and commonly used.Standard techniques can be used for chemosynthesis, the experimenter is prepared and send and treated to chemical analysis, at medication preparation.Some this technology and method can for example find in the following document: " Carbohydrate Modifications inAntisense Research " Sangvi and Cook (editor), American ChemicalSociety, Washington D.C., 1994; With " Remington ' s PharmaceuticalSciences, " Mack Publishing Co., Easton, Pa., 18th edition, 1990, these documents are that any purpose is attached to herein by reference.Under situation about allowing, present disclosure is all patents, patent application, disclosed application and other publications of indication and GenBank and other data bases' sequence in the whole text, and all integral body is attached to herein by reference.

Following term has following implication, unless otherwise:

Term used herein " nucleoside " means the glycosylamine that comprises nuclear base and sugar.Nucleoside includes but not limited to natural nucleus glycoside, dealkalize base (abasic) nucleoside, modified nucleoside and has mimic base and/or the nucleoside of glycosyl group.

Term used herein " nucleotide " refers to comprise the nuclear base and sugar reaches and the glycosylamine of the bound phosphate groups that sugar is covalently bound.Nucleotide can be with multiple substituent any modification.

Term used herein " nuclear base " refers to the base portion of nucleoside or nucleotide.The nuclear base can comprise any hydrogen-bonded atom or atomic group can take place with the base of another nucleic acid.

Term used herein " heterocyclic base moiety " refers to comprise heterocyclic nuclear base.

Term used herein " deoxyribonucleotide " means the nucleotide that has hydrogen in 2 ' position of the sugar moieties of nucleotide.Deoxyribonucleotide can be with multiple substituent any modification.

Term used herein " ribonucleotide " means the nucleotide that has hydroxyl in 2 ' position of the sugar moieties of nucleotide.Ribonucleotide can be with multiple substituent any modification.

Term used herein " oligomeric compounds " refer to comprise two or more substructures (sub-structure) and can with the poly structure of certain area hybridization of nucleic acid molecules.In certain embodiments, oligomeric compounds is an oligonucleoside.In certain embodiments, oligomeric compounds is an oligonucleotide.In certain embodiments, oligomeric compounds is an antisense compounds.In certain embodiments, oligomeric compounds is an antisense oligonucleotide.In certain embodiments, oligomeric compounds is short antisense compounds.In certain embodiments, oligomeric compounds is short antisense oligonucleotide.In certain embodiments, oligomeric compounds is a chimeric oligonucleotide.

Term used herein " monomer " refers to the single unit of oligomer.Monomer includes but not limited to nucleoside and nucleotide, no matter is natural or modified.

" oligonucleoside " used herein wherein connects the oligonucleotide that key does not contain phosphorus atoms between nucleoside.

Term used herein " oligonucleotide " refers to comprise the oligomeric compounds of a plurality of nucleotide that link together.In certain embodiments, the one or more nucleotide in the oligonucleotide are modified.In certain embodiments, oligonucleotide comprises ribonucleic acid (RNA) or DNA (deoxyribonucleic acid) (DNA).In certain embodiments, oligonucleotide is made up of natural and/or non-natural nuclear base, sugar and covalency internucleotide linkage (internucleotide linkage), and also can comprise non-nucleic acid conjugate (conjugate).

" internucleotide linkage " used herein refers to that the covalency between the adjacent nucleotide connects key.

Term used herein " Dan Julian key " refers to that two covalency between the monomer connect key.The Dan Julian key includes but not limited to connect key between internucleotide linkage and nucleoside.

" connecting key between natural nucleotide " used herein refers to that 3 '-5 ' di-phosphate ester connects key.

Term used herein " antisense compounds " refers to the target nucleic acids molecule of hybridizing with it complementary oligomeric compounds to the small part.In certain embodiments, antisense compounds can be regulated the expression of (improve or reduce) target nucleic acids.Antisense compounds includes but not limited to following chemical compound: the chimeric combination of oligonucleotide, oligonucleoside, oligonucleotide analogs, oligonucleotide mimetic and these chemical compounds (chimeric combination).Therefore, though all antisense compounds all are oligomeric compounds, not every oligomeric compounds all is an antisense compounds.

Term used herein " antisense oligonucleotide " refers to belong to the antisense compounds of oligonucleotide.

Term used herein " parent antisense oligonucleotide " refers to have the oligonucleotide deoxidation interval region, 20 nucleotide of length, this deoxidation interval region has 10 2 '-deoxyribonucleotides, both sides respectively are connected to first and second pterions, each pterion has 52 '-O-(2-methoxy ethyl) ribonucleotides (5-10-5MOE gapmer), and this oligonucleotide comprises the sequence of corresponding short antisense compounds as parent.

Term used herein " short antisense compounds " refers to about 8,9,10,11,12,13,14,15 or 16 the monomeric antisense compounds of length.In certain embodiments, lack antisense compounds and have at least one high-affinity modification.

Term used herein " short antisense oligonucleotide " refers to the antisense oligonucleotide of about 8,9,10,11,12,13,14,15 or 16 nucleotide of length.In certain embodiments, lack antisense oligonucleotide and have at least one high-affinity modification.

Term used herein " short gapmer " refers to so short antisense oligonucleotide, and it has length is 2-14 the interval region of examining base for first and second pterions of 1-3 nucleotide and length independently separately.

Term used herein " die body " refers to the pattern (pattern) of not modified and modified nucleotide in the short antisense compounds.

Term used herein " chimeric antisense oligomers " refers to such antisense oligomeric compounds, connect between its at least one sugar, nuclear base or nucleoside the suffered modification of key with its in the middle of at least one other sugar, nuclear base or nucleoside between connect key and compare variant.Connecting key between remaining sugar, nuclear base and nucleoside can be modified or do not modified independently, identical or different.

Term used herein " chimeric antisense oligonucleotide " refers to such antisense oligonucleotide, connect between its at least one sugar, nuclear base or nucleoside the suffered modification of key with its in the middle of at least one other sugar, nuclear base or nucleoside between connect key and compare variant.Connecting key between remaining sugar, nuclear base and nucleoside can be modified or do not modified independently, identical or different.

Term used herein " mixed matrix antisense oligonucleotide " refers to such antisense oligonucleotide, has wherein that to connect at least one other internucleotide linkage of key and its between at least one nucleoside different.

Term used herein " target " refers to the protein that need regulate.

Term used herein " target gene " refer to the to encode gene of target.

Term used herein " target nucleic acids " and the nucleic acid molecules of target " coding " refer to its expression or the active any nucleic acid molecules that can regulate by antisense compounds.Target nucleic acids includes but not limited to from the RNA (including but not limited to pre-mRNA and mRNA or their part) that transcribes of DNA of coding target, and spreads out from the cDNA of this RNA and miRNA.For example, target nucleic acids can be that it expresses the cytogene relevant with specific disease or the morbid state mRNA of this genetic transcription (or from), perhaps from the nucleic acid molecules of infectious agent.

Term used herein " targeting " refers to the association in the specific nucleotide zone in the middle of antisense compounds and particular target nucleic acid molecules or the target nucleic acids molecule.

Term used herein " 5 ' target site " refer in the target nucleic acids with the complementary nucleotide of 5 ' of specific antisense compounds-least significant end (most) nucleotide.

Term used herein " 3 ' target site " refer in the target nucleic acids with the complementary nucleotide of 3 ' of specific antisense compounds-least significant end (most) nucleotide.

Term used herein " target zone (target region) " refer in the target molecules with the complementary part of one or more antisense compounds.

Term used herein " target section (target segment) " refers to the more fraction or the inferior part in certain zone in the middle of the target nucleic acids.

Term used herein " nuclear base complement " refers to and can examine the nuclear base that base is carried out base pairing with another.For example, in DNA, adenine (A) and thymus pyrimidine (T) complementation.For example, in RNA, adenine (A) and uracil (U) complementation.In certain embodiments, complementary nuclear base refers to the nuclear base that can carry out base pairing with the nuclear base of its target nucleic acids in the antisense compounds.For example, if the nuclear base of certain position of antisense compounds can with the nuclear base generation hydrogen bonding of certain position of target nucleic acids, think that then the hydrogen-bonded position between this oligonucleotide and this target nucleic acids is complementary for this nuclear base pair.

Term used herein " incomplementarity nuclear base " refers to a pair of like this nuclear base, and they can not form hydrogen bond mutually or otherwise support hybridization.

Term used herein " complementation " refers to that oligomeric compounds is by examining the ability of base complement and another oligomeric compounds or nucleic acid hybridization.In certain embodiments, can stablize associating nuclear base between antisense compounds and the target thereof when occupied when relevant position that antisense compounds and its target have sufficient amount separately in the molecule is made by bonding mutually, antisense compounds and target thereof are just complementary mutually.Those of skill in the art will recognize that to comprise and do not eliminate the mispairing that oligomeric compounds keeps the ability of association state, this is possible.Therefore, antisense compounds described herein can comprise about 20% mispairing nucleotide (promptly not being and the complementary nuclear base of the corresponding nucleotide of target) at most.Preferably, antisense compounds contain be no more than about 15%, more preferably no more than about 10%, be most preferably not exceeding 5% mispairing or do not contain mispairing.Remaining nucleotide is complementary nuclear base, otherwise is exactly the nuclear base (for example universal base) that can not destroy hybridization.Those of ordinary skills will appreciate that, chemical compound that this paper provided and target nucleic acids 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% complementation.

Term used herein " mispairing " refers to the incomplementarity nuclear base in the middle of the complementary oligomeric compounds.

" hybridization " used herein means the pairing of complementary oligomeric compounds (for example antisense compounds and its target nucleic acids).Though do not want to be confined to concrete mechanism, prevailing pairing mechanism relates to the hydrogen bonding between complementary nucleoside base or the nucleotide base (nuclear base), and it can be Watson-Crick, Hoogsteen or trans Hoogsteen hydrogen bonding.For example, natural bases adenine is and natural nucleus base thymidine and the complementary nuclear base of uracil that they match by forming hydrogen bond.Natural base guanine is and natural base cytosine and the complementary nuclear base of 5-methylcytosine.Hybridization can occur under different situations.

This ability of the affinity that the affinity that term used herein " specific hybrid " refers to oligomeric compounds and the hybridization of nucleic acid site is hybridized greater than its and another nucleic acid site.In certain embodiments, antisense oligonucleotide and target locus specificity hybridization above one.

" design " used herein, " being designed to " refer to design can with the process of the oligomeric compounds of selected nucleic acid molecules specific hybrid.

Term used herein " adjusting " refers to compare with regulating preceding function or active level, and disturbance (perturbation) has taken place for function or activity.For example, regulate the variation comprise gene expression, this variation can be to improve (stimulate or induce) or reduce (suppress or reduce).As further example, the adjusting of expression comprises selects in addition disturbance to the splice site of pre-mRNA processing.

Term used herein " expression " refers to that all convert the encoding information of gene to the function and the step of the structure that exists and operate according to this in cell.This structure includes but not limited to the product transcribing and translate.

" variant " used herein refers to can be from the alternative rna transcription thing of identical DNA genome area generation.Variant includes but not limited to " pre-mRNA variant ", and this transcript and other transcript differences that produce from identical genomic DNA are their initial or final position, and this transcript contains intron sequences and exon sequence.Variant also includes but not limited to have alternately splice junction or alternately those variants of initial sum termination codon.

" through the monomer of high-affinity modification " used herein refers to such monomer, it is compared with natural monomer to have between at least one modified nuclear base, nucleoside and connects key or sugar moieties, and the result should modify to improve and comprise through the monomeric antisense compounds of high-affinity modification and the affinity of its target nucleic acids.High-affinity is modified the monomer (for example nucleoside and nucleotide) that includes but not limited to comprise through the sugar of 2 '-modification.

Term used herein " through 2 '-modify " or " 2 '-replace " mean the substituent sugar that comprises in 2 ' position outside H or the OH.Monomer through 2 '-modification includes but not limited to have such as 2 ' following-substituent BNA and monomer (for example nucleoside and nucleotide): pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) or O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, lacking antisense compounds comprises and does not have formula 2 '-O (CH₂)_n2 ' of H-modification monomer, wherein n is 1-6.In certain embodiments, lacking antisense compounds comprises and does not have formula 2 '-OCH₃2 '-modify monomer.In certain embodiments, lacking antisense compounds comprises and does not have formula 2 '-O (CH₂)₂OCH₃2 '-modify monomer.

Term used herein " dicyclo nucleic acid " or " BNA " or " bicyclic nucleoside " or " dicyclic ring thuja acid " refer to such nucleoside or nucleotide, wherein the furanose of nucleoside partly comprises the abutment that connects two carbon atoms on the furanose ring, thereby forms the dicyclo ring system.

Term used herein " methylene oxygen base BNA " refers to β-D-methylene oxygen base BNA alone, unless otherwise.

Term used herein " MOE " refers to 2 '-methoxy ethyl substituent group.

Term used herein " gapmer " refers to so chimeric oligomeric compounds, the zone (" pterion ") that it comprises middle section (" (gap) at interval ") and is positioned at the middle section both sides, wherein this comprises at least one modification different with the modification in each pterion at interval.This modification comprises that nuclear base modification, Dan Julian key (monomeric linkage) are modified and is sugar-modified and do not have a modification (not modified).Therefore, in certain embodiments, the nucleotide in each pterion connects key and nucleotide at interval, and to connect key different.In certain embodiments, each pterion comprises the nucleotide that has through the high-affinity modification, then comprises at interval not have this modified nucleotide.In certain embodiments, the nucleotide in and the nucleotide in the pterion all comprise the high-affinity modification at interval, but the modification of the high-affinity is at interval modified different with the high-affinity in the pterion.In certain embodiments, the modification in the pterion is identical mutually.In certain embodiments, the modification in the pterion is different mutually.In certain embodiments, the nucleotide in is not modified at interval, and the nucleotide in the pterion is modified.In certain embodiments, the modification in each pterion is identical.In certain embodiments, the modification in pterion is different with the modification in another pterion.In certain embodiments, lacking antisense compounds is the gapmer that has 2 '-deoxyribonucleotide and have the nucleotide of modifying through high-affinity in the interval in the pterion.

Term used herein " prodrug " refers to such therapeutic agent, and it prepares with inactive form, and the effect by endogenous enzymes or other chemical substances and/or condition in the middle of health or its cell changes into activity form (being medicine).

Term used herein " the acceptable salt of medicine " refers to the salt of reactive compound, and this salt can keep the required biological activity of reactive compound, does not give unwanted toxicology effect to reactive compound again.

Term used herein " cap " or " distal end cap part " refer to the chemical modification that the arbitrary end at antisense compounds mixes.

Term used herein " prevention " refers to the outbreak of disease or disease or development postponed or stops several hrs to several days, time of preferred several thoughtful some months.

Term used herein " improvement " refers to alleviate at least one indication (indicator) of disease or severity of disease.The seriousness indication can be measured by well known to a person skilled in the art subjectivity or objective measurement approach.

Term used herein " treatment " is showed and is given change or the improvement that compositions of the present invention realizes disease or disease.Prevention, improvement and/or treatment may require termly or give multidose before disease or disease outbreak, to change the process of disease or disease.In addition, single medicament can be used for single individuality, with sequential ground or side by side carry out each prevention, improvement and the treatment of disease or disease.

Term used herein " medicament " refers to can provide the material of curative effect when giving the experimenter.

Term used herein " treatment effective dose " refers to provide the amount of the medicament of curative effect to animal.

" giving " used herein refers to medicament is offered animal, includes but not limited to give to give with the oneself by the medical professional.

Term used herein " gives " to refer to give animal with two or more medicaments altogether.These two or more medicaments can be in single pharmaceutical composition, perhaps can be respectively in independent pharmaceutical composition.Each of these two or more medicaments can give by the identical or different approach that gives.Contain parallel giving or sequential giving altogether.

Term used herein " pharmaceutical composition " refers to be suitable for giving the mixture of each individual material.For example, pharmaceutical composition can comprise antisense oligonucleotide and aseptic aqueous solution.

Term used herein " individuality " refers to selected people or the non-human animal who treats and participate in therapy.

Term used herein " animal " refers to people or non-human animal, includes but not limited to mice, rat, rabbit, Canis familiaris L., cat, pig and non-human primate, includes but not limited to monkey and chimpanzee.

Term used herein " experimenter " refers to be given the animal of pharmaceutical composition, includes but not limited to the people.

Term used herein " persistent period " refers to the period that activity or incident continue.In certain embodiments, the treatment persistent period is the period that gives each dosage of medicament.

Term used herein " parenteral gives " refers to give by injection or infusion.Parenteral includes but not limited to subcutaneously give, intravenous gives or or intramuscular give.

Make a comment or criticism gives below skin term used herein " subcutaneous giving "." intravenous gives " means and is administered in the vein.

Term " dosage " used herein (dose) " specified amount of the medicament that refers in single gives to be provided.In certain embodiments, dosage can or repeatedly be injected agent (bolus), tablet or injection and gives with twice.For example, need in the subcutaneous embodiment that gives at some, the desired volume of required dosage is not easy to supply by single injection.In this embodiment, can adopt twice or multiple injection reaches required dosage.In certain embodiments, dosage can give with twice or multiple injection, so that the injection site reaction in the individuality minimizes.

Term " dosage " unit used herein (dosage unit) " refer to provide according to this form of medicament.In certain embodiments, dosage unit is the bottle (vial) that comprises freeze dried antisense oligonucleotide.In certain embodiments, dosage unit is the bottle that comprises (reconstituted) antisense oligonucleotide of redissolution.

Term used herein " medicament " refers to can provide the material of curative effect when giving individuality.For example, in certain embodiments, antisense oligonucleotide is a medicament.

Term used herein " active pharmaceutical ingredient " refers to the material that required effect can be provided in the pharmaceutical composition.

Term used herein " treatment effective dose " refers to provide the amount of the medicament of curative effect to individuality.In certain embodiments, the treatment effective dose of antisense compounds is to need to give the amount that is produced observable effects.

Term used herein " hypercholesterolemia " refers to the serum cholesterol rising to be the disease of feature.

Term used herein " hyperlipemia " refers to the serum lipids rising to be the disease of feature.

Term used herein " hypertriglyceridemia " refers to the triglyceride levels rising to be the disease of feature.

Term used herein " non-familial hypercholesterolemia " refers to that not to be that the result's of single genetic gene mutation cholesterol raises be the disease of feature.

Term used herein " polygenes hypercholesterolemia " refers to that it is the disease of feature that the cholesterol that causes with the influence by multiple inherited genetic factors raises.In certain embodiments, the polygenes hypercholesterolemia can be by the meals of lipid picked-up aggravation.

Term used herein " familial hypercholesterolemia (FH) " refers to LDL-receptor (LDL-R) gene mutation, LDL-C significantly raises and atherosclerosis outbreak too early is the autosomal dominant metabolic disease of feature.When individuality meets one of following standard or make a definite diagnosis the generation familial hypercholesterolemia when multinomial: genetic test is confirmed the LDL-acceptor gene of 2 sudden changes; Genetic test is confirmed the LDL-acceptor gene of 1 sudden change; Medical history record has the LDL-cholesterol of the untreated 500mg/dL of surpassing; Xanthoma tendinosum and/or xanthomata cutis took place before 10 years old; Perhaps parents have the record that the serum LDL-cholesterol before the lipid consistent with the heterozygous familial hypercholesterolemia reduces therapy raises.

Term used herein " familial hypercholesterolemia isozygotys " or " HoFH " refer to have with father and mother both sides' LDL-R gene the disease of the feature of sporting.

Term used herein " heterozygous familial hypercholesterolemia " or " HeFH " refer to that the LDL-R gene with father and mother one side has the disease of the feature of sporting.

It is the disease of feature that term used herein " Combination dyslipidemia " refers to raise with serum cholesterol rising and serum triglycerides.

It is the disease of feature that term used herein " diabetic dyslipidemia " or " type ii diabetes companion dyslipidemia " refer to increase with type ii diabetes, HDL-C minimizing, serum triglycerides rising and small and dense LDL granule.

What term used herein " CHD risk equipotential outbreak (risk equivalent) " referred to clinical atheromatosis brings high risk indication to coronary heart disease.For example, in certain embodiments, the outbreak of CHD risk equipotential includes but not limited to the aneurysm of clinical crown cardiopathia, symptom type carotid disease, peripheral arterial disease and/or abdominal aorta.

Term used herein " non-alcoholic fatty liver disease (NAFLD) " refers to that the liver fat inflammation not cause because of abuse of alcohol (for example alcohol consumption was above 20g/ days) is the disease of feature.In certain embodiments, NAFLD is relevant with insulin resistance and metabolism syndrome.

Term used herein " non-alcoholic stellato-hepatitis (NASH) " refers to that with the inflammation that does not cause because of alcohol abuse in the liver and fat and fibrosis tissue accumulation be the disease of feature.

NASH is the extreme form of NAFLD.

Term used herein " major risk factors " refers to facilitate the high risk factor of specified disease or disease.In certain embodiments, the major risk factors of coronary heart disease includes but not limited to smoking, hypertension, low HDL-C, familial history of coronary artery disease and age.

Term used herein " CHD risk factor " refers to outbreak of CHD risk equipotential and major risk factors.

Term used herein " coronary heart disease (CHD) " show heart blood supply and oxygen supply little blood vessel narrow this atherosclerotic often result.

Term used herein " coronary heart disease risk attenuating " refers to an attenuating of knowing from experience the probability of development coronary heart disease.In certain embodiments, the attenuating of coronary heart disease risk is that improvement (for example reduction of LDL-C level) by one or more CHD risk factor is measured.

Term used herein " atherosclerosis " refers to existing for of fatty deposits arteriosclerosis feature, that influence large artery trunks and medium-sized artery.Fatty deposits is called " tremulous pulse medicated porridge sample tumor (atheroma) " or " speckle (plaque) ", and it mainly is made up of cholesterol and other fat, calcium and scar tissue, can damage the tremulous pulse lining.

Term used herein " coronary disease medical history " refers to tangible clinically coronary heart disease, and a situation arises in individuality or individual family member's medical history.

Term used herein " coronary heart disease of early showing effect " refers to make a definite diagnosis coronary heart disease before 50 years old.

Term used herein " inhibin does not tolerate individuality " refers to such individuality, and creatine kinase increases because the inhibin therapy can experience for he, the liver function test is unusual, in myalgia or the central nervous system's side effect one or multinomial.

Term used herein " effect " refers to produce the ability of required effect.For example, lipid reduces the attenuating that the effect of therapy can be in LDL-C, VLDL-C, IDL-C, non-HDL-C, ApoB, lipoprotein (a) or the triglyceride or multinomial concentration.

" acceptable security feature (safety profile) refers to the side effect pattern in the middle of clinical acceptable limit to term used herein.

Term used herein " side effect " refers to the physiological reaction outside the effect that be attributable to treat, required.In certain embodiments, side effect includes but not limited to that the test of injection site reaction, liver function is unusual, renal dysfunction, hepatotoxicity, Toxicity of Kidney, the central nervous system is unusual and myopathy.For example, the raising of transamination enzyme level can be represented hepatotoxicity or abnormal liver function in the serum.For example, the bilirubin raising can be represented hepatotoxicity or abnormal liver function.

Term used herein " injection site reaction " refers to the chafing of individual injection site or rubescent unusually.

Term used herein " individual compliance " refer to individual to recommend or the adhering to of specified therapy.

Term used herein " lipid reduction therapy " refers to offer individual to reduce the therapeutic scheme of one or more lipids in the individuality.In certain embodiments, provide lipid to reduce therapy to reduce among ApoB in the individuality, T-CHOL, LDL-C, VLDL-C, IDL-C, non-HDL-C, triglyceride, small and dense LDL granule and the Lp (a) one or more.

Term used herein " lipid reduction medicament " refers to offer individual medicament with the reduction that realizes the lipid in the individuality.For example, in certain embodiments, lipid is reduced medicament offer individual to reduce in ApoB, LDL-C, T-CHOL and the triglyceride one or more.

Term used herein " LDL-C target " refers to that lipid reduces needed LDL-C level after the therapy.

Term used herein " is comply with " and is referred to that individuality adheres to the therapy of being recommended.

Term used herein " therapy of being recommended " refers to that the medical professional recommends to treat, improvement or prophylactic therapeutic scheme.

Term used herein " low LDL-receptor active " refers to the enough not high LDL-receptor active that must keep the clinically acceptable level of LDL-C in blood flow.

Term used herein " cardiovascular consequence " refers to the appearance of great disadvantageous cardiovascular event.

Term used herein " improved cardiovascular consequence " refers to the minimizing of the appearance of great disadvantageous cardiovascular event or its risk.The example of great disadvantageous cardiovascular event includes but not limited to death, re-infarction, apoplexy, cardiogenic shock, pulmonary edema, asystole and atrial arrhythmia.

Term used herein " surrogate markers of cardiovascular consequence " refers to the indirect indication of cardiovascular event or its risk.For example, the surrogate markers of cardiovascular consequence comprises carotid intimal medial thickness (CIMT).Another example of the surrogate markers of cardiovascular consequence comprises tremulous pulse medicated porridge sample tumor size.The big I of tremulous pulse medicated porridge sample tumor is measured by intravascular ultrasound (IVUS).

Term used herein " HDL-C raising " refers to Serum HDL-C raising in time in the individuality.

Term used herein " lipid reduction " refers to one or more serum lipids reduction in time in the individuality.

Term used herein " metabolic disease " refers to that with the change of metabolic function or disturbance be the disease of feature." metabolic " and " metabolism " is term well known in the art, generally comprises the whole biochemical process that occurs in the middle of the live organism.Metabolic disease includes but not limited to hyperglycemia, prediabetes, diabetes (I type and II type), obesity, insulin resistance and metabolism syndrome.

Term used herein " metabolism syndrome " refers to metabolic lipid of originating from of a monoid and non-lipid cardiovascular risk factors.It and vague generalization (generalized) metabolic disease that is called insulin resistance are closely linked.U.s. national cholesterol education program (National Cholesterol Education Program, NCEP) (Adult Treatment Panel III ATPIII) has set up the standard of diagnosing metabolism syndrome when having three or more to exist in five risk determiners to adult treatment expert scheme III.These five risk determiners are, with the abdominal obesity of waistline definition, the man is greater than 102cm, and the woman is greater than 88cm; Triglyceride levels is more than or equal to 150mg/dL; The HDL cholesterol levels, the man is less than 40mg/dL, and the woman is less than 50mg/dL; Blood pressure is more than or equal to 130/85mm Hg; With the fasting glucose level more than or equal to 110mg/dL.These determiners in clinical practice, can measure easily (JAMA, 2001,285:2486-2497).

Term used herein " alkyl " refers to contain the saturated straight chain or the branched-chain hydrocarbons group of maximum 24 carbon atoms.The example of alkyl group includes but not limited to methyl, ethyl, propyl group, butyl, isopropyl, n-hexyl, octyl group, decyl, dodecyl etc.Alkyl group generally includes 1 to about 24 carbon atoms, and more generally 1 to about 12 carbon atom (C₁-C₁₂Alkyl), more preferably 1 to about 6 carbon atoms.Term used herein " low alkyl group " comprises 1 to about 6 carbon atoms.Alkyl group used herein can be chosen wantonly and comprise one or more other substituent groups.

Saturated straight chain or side chain hydrocarbon chain group that term used herein " thiazolinyl " refers to contain maximum 24 carbon atoms and has at least one carbon-to-carbon double bond.The example of alkenyl group includes but not limited to vinyl, acrylic, cyclobutenyl, 1-methyl-2-butene-1-base, diene such as 1,3-butadiene etc.Alkenyl group generally includes 2 to about 24 carbon atoms, and more generally 2 to about 12 carbon atoms, and more preferably 2 to about 6 carbon atoms.Alkenyl group used herein can be chosen wantonly and comprise one or more other substituent groups.

Saturated straight chain or branched-chain hydrocarbons group that term used herein " alkynyl " refers to contain maximum 24 carbon atoms and has at least one carbon-to-carbon triple bond.The example of alkynyl includes but not limited to acetenyl, 1-propinyl, ethyl acetylene base etc.Alkynyl group generally includes 2 to about 24 carbon atoms, and more generally 2 to about 12 carbon atoms, and more preferably 2 to about 6 carbon atoms.Alkynyl group used herein can be chosen wantonly and comprise one or more other substituent groups.

Term used herein " aminoalkyl " refers to the amino alkyl group that replaces.This term mean be included in any position have amino substituent group and wherein alkyl group connect the C of key by amino group and parent molecule₁-C₁₂Alkyl group.The moieties of aminoalkyl groups and/or amino part can further be substituted base and replace.

Term used herein " aliphatic series " refers to contain maximum 24 carbon atoms, wherein the saturation between any two carbon atoms is singly-bound, two key or triple-linked straight or branched hydrocarbyl group.Aliphatic group preferably includes 1 to about 24 carbon atoms, and more generally 1 to about 12 carbon atoms, and more preferably 1 to about 6 carbon atoms.The straight or branched of aliphatic group can be interrupted by one or more hetero atoms of nitrogen, oxygen, sulfur and phosphorus that comprise.This aliphatic group that is interrupted by hetero atom includes but not limited to poly-alkoxyl (as the polyoxygenated alkylene), polyamine and poly-imines.Aliphatic group used herein can be chosen wantonly and comprise other substituent group.

Term " alicyclic ring " or " alcyl " refer to that ring wherein belongs to aliphatic ring system.This ring system can comprise one or more rings, and wherein at least one ring is aliphatic.Preferred alicyclic ring comprises wherein having about 5 rings to about 9 carbon atoms.Alicyclic ring used herein can be chosen wantonly and comprise other substituent group.

Term used herein " alkoxyl " refers to the group that forms between alkyl group and oxygen atom, wherein this oxygen atom is in order to be connected alkoxy base with parent molecule.The example of alkoxyl includes but not limited to methoxyl group, ethyoxyl, propoxyl group, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentyloxy, neopentyl oxygen, positive hexyloxy etc.Alkoxy base used herein can be chosen wantonly and comprise other substituent group.

Term used herein " halogen " and " halogen " refer to be selected from the atom of fluorine, chlorine, bromine and iodine.

Term used herein " aryl " and " aromatics " refer to have the monocycle or the multi-ring carbon-loop system group of one or more aromatic rings.The example of aromatic yl group includes but not limited to phenyl, naphthyl, tetralyl, indanyl, indenyl (idenyl) etc.The preferred aryl groups ring system has about 5 to about 20 carbon atoms in one or more rings.Aromatic yl group used herein can be chosen wantonly and comprise other substituent group.

Term used herein " aralkyl " and " aryl alkyl " refer to the group that forms between alkyl group and aromatic yl group, wherein alkyl group is in order to be connected aromatic alkyl group with parent molecule.Example includes but not limited to benzyl, phenethyl etc.Aromatic alkyl group used herein can be chosen wantonly and comprise the other substituent group that is connected with alkyl group, aromatic yl group or this two groups of constituting described group.

Term used herein " heterocyclic group " refers to such basic (radical) monocycle or polycyclic aromatic ring system, and it comprises at least one hetero atom and be undersaturated, fractional saturation or fully saturated, thereby comprises heteroaryl groups.Heterocyclic group also means and comprises such condensed ring system, and wherein one or more condensed ring contain at least one hetero atom, and other ring can contain one or more hetero atoms or the optional hetero atom that do not contain.Heterocyclic group generally includes at least one atom that is selected from sulfur, nitrogen or oxygen.The example of heterocyclic group comprises [1,3] dioxolanes, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidyl, piperazinyl, oxazolidinyl, isoxazole alkyl, morpholinyl, thiazolidinyl, isothiazole alkyl, quinoxalinyl, pyridazine ketone group, tetrahydrofuran base etc.Heterocyclic group used herein can be chosen wantonly and comprise other substituent group.

Term used herein " heteroaryl " and " heteroaromatic " refer to comprise the group of monocycle or multi-ring aromatic ring ring system or condensed ring system, wherein at least one ring be aromatics and comprise one or more hetero atoms.Heteroaryl also means and comprises the condensed ring system, comprises that wherein one or more condensed ring do not contain heteroatomic system.Heteroaryl groups generally includes an annular atoms that is selected from sulfur, nitrogen or oxygen.The example of heteroaryl groups includes but not limited to pyridine radicals, pyrazinyl, pyrimidine radicals, pyrrole radicals, pyrazolyl, imidazole radicals, thiazolyl, oxazolyl, isoxazolyl, thiadiazolyl group, oxadiazole base, thienyl (thiophenyl), furyl, quinolyl, isoquinolyl, benzimidazolyl, benzoxazolyl, quinoxalinyl etc.Heteroaryl groups can be directly or is connected with parent molecule by coupling part such as aliphatic group or hetero atom.Heteroaryl groups used herein can be chosen wantonly and comprise other substituent group.

" heteroaryl alkyl " used herein refers to have the heteroaryl groups that the front defines of alkyl group, and this alkyl group can be connected the heteroaryl alkyl group with parent molecule.Example includes but not limited to pyridylmethyl, pyrimidinylethyl, naphthyridinyl propyl group etc.Heteroaryl alkyl group used herein can be chosen in heteroaryl moieties or moieties one or comprise other substituent group on two simultaneously wantonly.

Term used herein " monocycle or multiring structure " comprises all monocycles or polycyclic ring system, wherein multi-ring have a ring that condenses or link together, and this term means and comprises the monocycle that is selected from aliphatic group, alicyclic group, aryl, heteroaryl, aralkyl, aryl alkyl, heterocyclic group, heteroaryl, heteroaromatic group, heteroaryl alkyl separately and mix ring system.This monocycle or multiring structure can contain the ring that the same ring or degree of saturation have nothing in common with each other, and comprise saturated fully, fractional saturation or fully unsaturated.Each ring can comprise the ring of annular atoms to produce heterocycle and only to comprise the C annular atoms that is selected from C, N, O and S, this can exist in the mixing die body (mixied motif) of for example benzimidazole, one of them ring only has carboatomic ring atom, and condensed ring then has two nitrogen-atoms.Monocycle or multiring structure can further be substituted base and replace, phthalimide for example, and it has two=O group and is connected with one of them ring.In yet another aspect, monocycle or multiring structure can directly pass through annular atoms, be connected with parent molecule by substituent group or difunctional coupling part.

Term used herein " acyl group " refers to that organic acid removes the formed group of hydroxyl, and it has general formula-C (O)-X, wherein X normally aliphatic series, alicyclic or aromatics.Example comprises aliphatic carbonyl, aromatics carbonyl, aliphatic sulfonyl, aromatics sulfinyl, aliphatic sulfinyl, aromatics phosphoryl (aromatic phosphate), aliphatic phosphoryl (aliphatic phosphate) etc.Carboxyl groups used herein can be chosen wantonly and comprise other substituent group.

Term " alkyl " comprises the group that comprises C, O and H.Comprise straight chain group, branched group and cyclic group with any degree of saturation.This hydrocarbyl group can comprise the hetero atom of one or more N of being selected from, O and S, and can be further by the single replacement of one or more substituent groups or polysubstituted.

Term used herein " substituent group " comprises and is added to other groups or parent compound usually to strengthen required characteristic or to produce the group of required effect.Substituent group can be protected or not protected, and of can be added in the parent compound can add angle of striking or a plurality of angle of striking that adds.Substituent group also can further be replaced by other substituent groups, and can directly or by the linking group such as alkyl group or hydrocarbyl group be connected with parent compound.This substituent group includes but not limited to halogen, hydroxyl, alkyl, thiazolinyl, alkynyl, acyl group (C (O) R_Aa), carboxyl (C (O) O-R_Aa), aliphatic group, alicyclic group, alkoxyl, substituted oxo base (O-R_Aa), aryl, aralkyl, heterocyclic radical, heteroaryl, heteroaryl alkyl, amino (NR_BbR_Cc), imino group (=NR_Bb), amide groups (C (O) NR_BbR_CcOr-N (R_Bb) C (O) R_Aa), azido (N₃), nitro (NO₂), cyano group (CN), carbamoyl (OC (O) NR_BbR_CcOr-N (R_Bb) C (O)-OR_Aa), urea groups (N (R_Bb) C (O) NR_BbR_Cc), ghiourea group (N (R_Bb) C (S) NR_BbR_Cc), guanidine radicals (N (R_Bb) C (=NR_Bb) NR_BbR_Cc), amidino groups (C (=NR_Bb) NR_BbR_CcOr-N (R_Bb) C (NR_Bb) R_Aa), mercaptan (SR_Bb), sulfinyl (S (O) R_Bb), sulfonyl (S (O)₂R_Bb), sulfoamido (S (O)₂NR_BbR_CcOr-N (R_Bb) S (O)₂R_Bb) and conjugated group.Each R wherein_Aa, R_BbAnd R_CcIndependent is H, the optional chemical functional group who connects or other substituent group, and preferred list includes but not limited to H, alkyl, thiazolinyl, alkynyl, aliphatic group, alkoxyl, acyl group, aryl, aralkyl, heteroaryl, alicyclic group, heterocyclic group and heteroaryl alkyl.

B. some oligomeric compounds

In certain embodiments, than natural oligomer such as DNA or RNA, need carry out chemical modification to oligomeric compounds.Some this modification can change the activity of oligomeric compounds.Some this chemical modification can for example change activity in the following manner: improve the affinity of antisense compounds to its target nucleic acids, improve it to the resistance of one or more nucleases and/or the pharmacokinetics or the tissue distribution of change oligomeric compounds.In some cases, adopt chemical method to improve the affinity of oligomeric compounds, can make and to use short oligomeric compounds its target.

1. some monomer

In certain embodiments, oligomeric compounds comprises one or more modified monomers.In certain embodiments, oligomeric compounds comprises one or more high-affinity monomers.In certain embodiments, this high-affinity monomer is selected from the monomer (for example nucleoside and nucleotide) that comprises through the sugar of 2 '-modification, includes but not limited to have such as 2 ' following-substituent BNA and monomer (for example nucleoside and nucleotide): pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) or O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.

In certain embodiments, include but not limited to that short antisense compounds comprises one or more high-affinity monomers at interior oligomeric compounds of the present invention, condition is that this oligomeric compounds does not include and comprises 2 '-O (CH₂)_nThe nucleotide of H, wherein n is 1-6.

In certain embodiments, include but not limited to that short antisense compounds comprises one or more high-affinity monomers at interior oligomeric compounds of the present invention, condition is that this oligomeric compounds does not include and comprises 2 '-OCH₃Or 2 '-O (CH₂)₂OCH₃Nucleotide.

In certain embodiments, include but not limited to that short antisense compounds comprises one or more high-affinity monomers at interior oligomeric compounds of the present invention, condition is that this oligomeric compounds does not include α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA.

In certain embodiments, include but not limited to that short antisense compounds comprises one or more high-affinity monomers at interior oligomeric compounds of the present invention, condition is that this oligomeric compounds does not include β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA.

In certain embodiments, include but not limited to that short antisense compounds comprises one or more high-affinity monomers at interior oligomeric compounds of the present invention, condition is that this oligomeric compounds does not include α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA or β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA.

A. some examines base

The trona base section of nucleoside is heterocyclic base normally.This heterocyclic base of prevailing two classes is purine and pyrimidine.For the nucleoside that comprises furan pentose, phosphate group can be connected with 2 ', 3 ' or 5 ' hydroxylic moiety of sugar.When forming oligonucleotide, these phosphate groups are together covalently bound mutually with adjacent nucleoside, form the linear polymerization chemical compound.In the middle of oligonucleotide, it is to form skeleton between the nucleotide of oligonucleotide that phosphate group is said to be usually.Natural even key or the skeleton of RNA and DNA are 3 '-5 ' phosphodiester bonds.

Except " not modified " or " natural " nuclear base such as purine are examined bases adenine (A) and guanine (G) and pyrimidine nuclear base thymus pyrimidine (T), cytosine (C) and uracil (U), well known to a person skilled in the art many modified nuclear bases or examine the base analogies also to be applicable to chemical compound described herein.In certain embodiments, modified nuclear base is structurally to examine the quite proximate nuclear base of base with parent, for example 7-deazapurine, 5-methylcytosine or G-pincers (G-clamp).In certain embodiments, nuclear base analogies comprise more complicated structure, for example three Huan phenoxazines nuclear base analogies.The method for preparing above-mentioned modified nuclear base is well known to a person skilled in the art.

B. some sugar

Oligomeric compounds provided herein can comprise one or more monomers with modified sugar moieties, comprises nucleoside or nucleotide.For example, the furanose ring of nucleoside can be modified in many ways, includes but not limited to add substituent group, and two non-geminal annular atoms bridge joints form dicyclo nucleic acid (BNA).

In certain embodiments, oligomeric compounds comprises one or more BNA monomers.In some this embodiment, BNA includes but not limited to (A) α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, (B) β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, (C) ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, (D) amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and (E) oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA, as shown in Figure 1.

Some BNA structure of Fig. 1

In certain embodiments, the BNA chemical compound includes but not limited to have the chemical compound of at least one abutment between 4 ' and 2 ' position of sugar, wherein each abutment independently comprise 1 or 2-4 independently be selected from-[C (R₁) (R₂)]_n-,-C (R₁)=C (R₂)-,-C (R₁)=N-,-C (=NR₁)-,-C (=O)-,-C (=S)-,-O-,-Si (R₁)₂-,-S (=O)_x-and-N (R₁)-linking group;

Wherein:

X is 0,1 or 2;

N is 1,2,3 or 4;

Each R₁And R₂Independent is H, protecting group, hydroxyl, C₁-C₁₂Alkyl, substituted C₁-C₁₂Alkyl, C₂-C₁₂Thiazolinyl, substituted C₂-C₁₂Thiazolinyl, C₂-C₁₂Alkynyl, substituted C₂-C₁₂Alkynyl, C₅-C₂₀Aryl, substituted C₅-C₂₀Aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C₅-C₇Alcyl, substituted C₅-C₇Alcyl, halogen, OJ₁, NJ₁J₂, SJ₁, N₃, COOJ₁, acyl group (C (=O)-H), substituted acyl group, CN, sulfonyl (S (=O)₂-J₁) or sulfinyl (sulfoxyl) (S (=O)-J₁); With

Each J₁And J₂Independent is H, C₁-C₁₂Alkyl, substituted C₁-C₁₂Alkyl, C₂-C₁₂Thiazolinyl, substituted C₂-C₁₂Thiazolinyl, C₂-C₁₂Alkynyl, substituted C₂-C₁₂Alkynyl, C₅-C₂₀Aryl, substituted C₅-C₂₀Aryl, acyl group (C (=O)-H), substituted acyl group, heterocyclic radical, substituted heterocyclic radical, C₁-C₁₂Aminoalkyl, substituted C₁-C₁₂Aminoalkyl or protecting group.

In one embodiment, each abutment of BNA chemical compound independently is-[C (R₁) (R₂)]_n-,-[C (R₁) (R₂)]_n-O-,-C (R₁R₂)-N (R₁)-O-or-C (R₁R₂)-O-N (R₁)-.In another embodiment, each described abutment independently is 4 '-CH₂-2 ', 4 '-(CH₂)₂-2 ', 4 '-(CH₂)₃-2 ', 4 '-CH₂-O-2 ', 4 '-(CH₂)₂-O-2 ', 4 '-CH₂-O-N (R₁)-2 ' and 4 '-CH₂-N (R₁)-O-2 '-, each R wherein₁Independent is H, protecting group or C₁-C₁₂Alkyl.

Some BNA has been produced and open (Singh et al., Chem.Commun., 1998,4,455-456 in patent documentation and scientific literature; Koshkin et al., Tetrahedron, 1998,54,3607-3630; Wahlestedt et al., Proc.Natl.Acad.Sci.U.S.A., 2000,97,5633-5638; Kumar et al., Bioorg.Med.Chem.Lett., 1998,8,2219-2222; WO 94/14226; WO 2005/021570; Singh et al., J.Org.Chem., 1998,63,10035-10039.Disclose BNA authorize United States Patent (USP) and the example of published application comprise for example United States Patent (USP) 7,053,207; 6,268,490; 6,770,748; 6,794,499; 7,034,133 and 6,525,191; And the early stage publication number 2004-0171570 of the U.S.; 2004-0219565; 2004-0014959; 2003-0207841; 2004-0143114 and 20030082807.

This paper also provides such BNA, and wherein 2 ' of ribose ring-hydroxyl is connected with 4 ' carbon atom of sugar ring, thereby forms methylene oxygen base (4 '-CH₂-O-2 ') Lian Jian, (summary is seen Elayadi et al., Curr.Opinion Invens.Drugs, 2001,2,558-561 to form the dicyclo sugar moieties; Braasch et al., Chem.Biol., 2001,8 1-7; With Orum et al., Curr.Opinion Mol.Ther., 2001,3,239-243; In addition referring to United States Patent (USP) 6,268,490 and 6,670,461).Connecting key can be the methylene (CH of bridge joint 2 ' oxygen atom and 4 ' carbon atom₂-) group, so dicyclo is partly used term methylene oxygen base (4 '-CH₂-O-2 ') BNA; As this position is the ethylidene group, then uses term ethyleneoxy group (4 '-CH₂CH₂-O-2 ') BNA (Singh et al., Chem.Commun., 1998,4,455-456:Morita et al., Bioorganic Medicinal Chemistry, 2003,11,2211-2226).Methylene (4 '-CH₂-O-2 ') BNA and other dicyclo sugar analogues demonstrate very high with duplex heat stability complementary DNA and RNA (Tm=+3-+10 ℃), to the stability of the circumscribed degraded of 3 '-nucleic acid and good dissolution characteristics.Comprise BNA existing description of potent and nontoxic antisense oligonucleotide (Wahlestedt et al., Proc.Natl.Acad.Sci.U.S.A., 2000,97,5633-5638).

Methylene oxygen base (the 4 '-CH that had discussed₂-O-2 ') BNA isomer is α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA has proved that it has splendid stability to 3 '-exonuclease.α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA be incorporated into and shown in active antisense gapmer of potent antisense and the chimera (Frieden et al., nucleic acid s Research, 2003,21,6365-6372).

Methylene oxygen base (4 '-CH₂-O-2 ') existing (the Koshkin et al. that describes of the synthetic and preparation of BNA monomer adenine, cytosine, guanine, 5-methyl-cytosine, thymus pyrimidine and uracil and their oligomerization and nucleic acid recognizing characteristic, Tetrahedron, 1998,54,3607-3630).BNA and preparation thereof also have description in WO 98/39352 and WO 99/14226.

Methylene oxygen base (4 '-CH₂-O-2 ') BNA analog thiophosphate-methylene oxygen base (4 '-CH₂-O-2 ') BNA and 2 '-sulfo--BNA also be produced (Kumar et al., Bioorg.Med.Chem.Lett., 1998,8,2219-2222).The relevant preparation that comprises as the locking nucleoside analog of the oligodeoxyribonucleotide duplex of the substrate of nucleic acid polymerase, also existing describe (Wengel et al., WO 99/14226).In addition, this novel conformation limited (comformationally restricted) high-affinity oligonucleotide analogs of 2 '-amino-BNA synthetic also has description (Singh et al. in the art, J.Org.Chem., 1998,63,10035-10039).In addition, 2 '-amino-BNA and 2 '-methylamino-BNA has also obtained preparation, existing report before the heat stability of the duplex of they and complementary RNA chain and DNA chain.

Modified sugar moieties is known, can be used to change the affinity of (normally improving) antisense compounds and its target and/or improves the nuclease resistance.The representative list of preferred modified sugar includes but not limited to comprise methylene oxygen base (4 '-CH through the sugar (BNA) of dicyclo modification₂-O-2 ') BNA and ethyleneoxy group (4 '-(CH₂)₂-O-2 ' abutment) BNA; Substituted sugar particularly has 2 '-F, 2 '-OCH₃Or 2 '-O (CH₂)₂-OCH₃Substituent 2 '-replacement sugar; Sugar with 4 '-thio-modification.Substituting of the also available sugar simulation of sugar group and so on.The method for preparing modified sugar is well known to a person skilled in the art.The patent and the prospectus of the preparation of this modified sugar of some representational instructions include but not limited to 4,981,957; 5,118,800; 5,319,080; 5,359,044; 5,393,878; 5,446,137; 5,466,786; 5,514,785; 5,519,134; 5,567,811; 5,576,427; 5,591,722; 5,597,909; 5,610,300; 5,627,053; 5,639,873; 5,646,265; 5,658,873; 5,670,633; 5,792,747; 5,700,920; 6,531,584; With 6,600,032; With WO 2005/121371.

In certain embodiments, BNA comprises the bicyclic nucleoside with following formula:

Wherein:

Bx is a heterocyclic base moiety;

T₁Be H or hydroxyl protecting group;

T₂Be H, hydroxyl protecting group or reactive phosphorus group;

Z is C₁-C₆Alkyl, C₂-C₆Thiazolinyl, C₂-C₆Alkynyl, substituted C₁-C₆Alkyl, substituted C₂-C₆Thiazolinyl, substituted C₂-C₆Alkynyl, acyl group, substituted acyl group or substituted amide.

In one embodiment, the independent quilt of each described substituent group independently is selected from the single replacement of following optional protected substituent group or polysubstituted: halogen, oxo base, hydroxyl, OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂And CN, wherein each J₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁

In some this embodiment, the independent quilt of each described substituent group independently is selected from the single replacement of following substituent group or polysubstituted: halogen, oxo base, hydroxyl, OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁And NJ₃C (=X) NJ₁J₂, each J wherein₁, J₂And J₃Independent is H, C₁-C₆Alkyl or substituted C₁-C₆Alkyl, X are O or NJ₁

In certain embodiments, the Z group is by one or more X^xThe C that replaces₁-C₆Alkyl, wherein each X^xIndependent is OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂Or CN; Each J wherein₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁In another embodiment, the Z group is by one or more X^xThe C that replaces₁-C₆Alkyl, wherein each X^xIndependent is halogen (for example fluorine), hydroxyl, alkoxyl (CH for example₃O-), substituted alkoxyl or azido.

In certain embodiments, the Z group is-CH₂X^x, X wherein^xBe OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂Or CN; Each J wherein₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁In another embodiment, the Z group is-CH₂X^x, X wherein^xBe halogen (for example fluorine), hydroxyl, alkoxyl (CH for example₃O-) or azido.

In some this embodiment, the Z group is (R)-configuration:

In some this embodiment, the Z group is (S)-configuration:

In certain embodiments, each T₁And T₂Be hydroxyl protecting group.The preferred list of hydroxyl protecting group comprises benzyl, benzoyl, 2,6-dichloro benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl, methanesulfonate, tosylate, dimethoxytrityl (DMT), 9-phenyl yanthine-9-base (Pixyl) and 9-(right-methoxyphenyl) xanthine-9-base (MOX).In certain embodiments, T₁For being selected from the hydroxyl protecting group of acetyl group, benzyl, t-butyldimethylsilyl, t-butyldiphenylsilyl and dimethoxytrityl, wherein preferred hydroxyl protecting group is T₁Be 4,4 '-dimethoxytrityl.

In certain embodiments, T₂Be the reactive phosphorus group, wherein preferred reactive phosphorus group comprises diisopropyl hydroxyl-oxethyl phosphoramidite and H-phosphate radical.In certain embodiments, T₁Be 4,4 '-dimethoxytrityl, T₂Be diisopropyl hydroxyl-oxethyl phosphoramidite.

In certain embodiments, oligomeric compounds has the monomer of at least one following formula:

Or the monomer of following formula:

Or the monomer of following formula:

Wherein

Bx is a heterocyclic base moiety;

T₃Be linking group, nucleotide, oligonucleoside, oligonucleotide, single poly-subunit or oligomeric compounds between H, hydroxyl protecting group, the connection conjugated group that is connected with nucleoside or nucleoside;

T₄Be linking group, nucleotide, oligonucleoside, the poly-subunit of oligonucleotide list or oligomeric compounds between H, hydroxyl protecting group, the connection conjugated group that is connected with nucleoside or nucleoside;

T wherein₃And T₄In linking group, nucleotide, oligonucleoside, oligonucleotide, single poly-subunit or oligomeric compounds between at least one is with nucleoside is connected nucleoside; With

Z is C₁-C₆Alkyl, C₂-C₆Thiazolinyl, C₂-C₆Alkynyl, substituted C₁-C₆Alkyl, substituted C₂-C₆Thiazolinyl, substituted C₂-C₆Alkynyl, acyl group, substituted acyl group or substituted amide.

In one embodiment, the independent quilt of each described substituent group independently is selected from the single replacement of following optional protected substituent group or polysubstituted: halogen, oxo base, hydroxyl, OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂And CN, wherein each J₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁

In one embodiment, the independent quilt of each described substituent group independently is selected from the single replacement of following substituent group or polysubstituted: halogen, oxo base, hydroxyl, OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁And NJ₃C (=X) NJ₁J₂, each J wherein₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O or NJ₁

In some this embodiment, at least one Z is C₁-C₆Alkyl or substituted C₁-C₆Alkyl.In certain embodiments, each Z independently is C₁-C₆Alkyl or substituted C₁-C₆Alkyl.In certain embodiments, at least one Z is C₁-C₆Alkyl.In certain embodiments, each Z independently is C₁-C₆Alkyl.In certain embodiments, at least one Z is a methyl.In certain embodiments, each Z is a methyl.In certain embodiments, at least one Z is an ethyl.In certain embodiments, each Z is an ethyl.In certain embodiments, at least one Z is substituted C₁-C₆Alkyl.In certain embodiments, each Z independently is substituted C₁-C₆Alkyl.In certain embodiments, at least one Z is substituted methyl.In certain embodiments, each Z is substituted methyl.In certain embodiments, at least one Z is substituted ethyl.In certain embodiments, each Z is substituted ethyl.

In certain embodiments, at least one substituent group is C₁-C₆(for example at least one Z is by one or more C to alkoxyl₁-C₆The C that alkoxyl replaces₁-C₆Alkyl).In another embodiment, each substituent group independently is C₁-C₆(for example each Z independently is by one or more C to alkoxyl₁-C₆The C that alkoxyl replaces₁-C₆Alkyl).

In certain embodiments, at least one C₁-C₆Alkoxy substituent is CH₃(for example at least one Z is CH to O-₃OCH₂-).In another embodiment, each C₁-C₆Alkoxy substituent is CH₃(for example each Z is CH to O-₃OCH₂-).

In certain embodiments, at least one substituent group is the halogen (C of at least one Z for being replaced by one or more halogens for example₁-C₆Alkyl).In certain embodiments, independently (for example each Z independently is the C that replaced by one or more halogens to each substituent group for halogen₁-C₆Alkyl).In certain embodiments, at least one halogenic substituent is that (for example at least one Z is CH to fluorine₂FCH₂-, CHF₂CH₂-or CF₃CH₂-).In certain embodiments, each halogenic substituent is that (for example each Z independently is CH to fluorine₂FCH₂-, CHF₂CH₂-or CF₃CH₂-).

In certain embodiments, at least one substituent group is the hydroxyl (C of at least one Z for being replaced by one or more hydroxyls for example₁-C₆Alkyl).In certain embodiments, independently (for example each Z independently is the C that replaced by one or more hydroxyls to each substituent group for hydroxyl₁-C₆Alkyl).In certain embodiments, at least one Z is HOCH₂-.In another embodiment, each Z is HOCH₂-.

In certain embodiments, at least one Z is CH₃-, CH₃CH₂-, CH₂OCH₃-, CH₂F-or HOCH₂-.In certain embodiments, each Z independently is CH₃-, CH₃CH₂-, CH₂OCH₃-, CH₂F-or HOCH₂-.

In certain embodiments, at least one Z group is by one or more X^xThe C that replaces₁-C₆Alkyl, wherein each X^xIndependent is OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂Or CN; Each J wherein₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁In another embodiment, at least one Z group is by one or more X^xThe C that replaces₁-C₆Alkyl, wherein each X^xIndependent is halogen (for example fluorine), hydroxyl, alkoxyl (CH for example₃O-) or azido.

In certain embodiments, each Z group independently is by one or more X^xThe C that replaces₁-C₆Alkyl, wherein each X^xIndependent is OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂Or CN; Each J wherein₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁In another embodiment, each Z group independently is by one or more X^xThe C that replaces₁-C₆Alkyl, wherein each X^xIndependent is halogen (for example fluorine), hydroxyl, alkoxyl (CH for example₃O-) or azido.

In certain embodiments, at least one Z group is-CH₂X^x, X wherein^xBe OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂Or CN; Each J wherein₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁In certain embodiments, at least one Z group is-CH₂X^x, X wherein^xBe halogen (for example fluorine), hydroxyl, alkoxyl (CH for example₃O-) or azido.

In certain embodiments, each Z group independently is-CH₂X^x, each X wherein^xIndependent is OJ₁, NJ₁J₂, SJ₁, N₃, OC (=X) J₁, OC (=X) NJ₁J₂, NJ₃C (=X) NJ₁J₂Or CN; Each J wherein₁, J₂And J₃Independent is H or C₁-C₆Alkyl, X are O, S or NJ₁In another embodiment, each Z group independently is-CH₂X^x, each X wherein^xIndependent is halogen (for example fluorine), hydroxyl, alkoxyl (CH for example₃O-) or azido.

In certain embodiments, at least one Z is CH₃-.In another embodiment, each Z is CH₃-.

In certain embodiments, at least one monomeric Z group is (the R)-configuration shown in the following formula:

Or (the R)-configuration shown in the following formula:

Or (the R)-configuration shown in the following formula:

In certain embodiments, the monomeric Z group of each of described formula is (R)-configuration.

In certain embodiments, at least one monomeric Z group is (the S)-configuration shown in the following formula:

Or (the S)-configuration shown in the following formula:

Or (the S)-configuration shown in the following formula:

In certain embodiments, the monomeric Z group of each of described formula is (S)-configuration.

In certain embodiments, T₃Be H or hydroxyl protecting group.In certain embodiments, T₄Be H or hydroxyl protecting group.In another embodiment, T₃For with nucleoside that nucleoside, nucleotide or single poly-subunit are connected between connect gene.In certain embodiments, T₄For with nucleoside that nucleoside, nucleotide or single poly-subunit are connected between linking group.In certain embodiments, T₃For with nucleoside that oligonucleoside or oligonucleotide are connected between linking group.In certain embodiments, T₄For with nucleoside that oligonucleoside or oligonucleotide are connected between linking group.In certain embodiments, T₃For with nucleoside that oligomeric compounds is connected between linking group.In certain embodiments, T₄For with nucleoside that oligomeric compounds is connected between linking group.In certain embodiments, T₃And T₄In at least one comprises linking group between the nucleoside that is selected from di-phosphate ester or thiophosphate.

In certain embodiments, oligomeric compounds has monomeric at least one zone of following formula of at least two adjacency:

Or monomeric at least one zone of the following formula of at least two adjacency:

Or monomeric at least one zone of the following formula of at least two adjacency:

In certain embodiments, oligomeric compounds comprises monomeric at least two zones of following formula of at least two adjacency.In certain embodiments, oligomeric compounds comprise interband every (gapped) oligomeric compounds.In certain embodiments, oligomeric compounds comprises about 8 to about 14 at least one zones in abutting connection with β-D-2 '-desoxyribofuranose yl nucleosides.In certain embodiments, oligomeric compounds comprises about 9 to about 12 at least one zones in abutting connection with β-D-2 '-desoxyribofuranose yl nucleosides.

In certain embodiments, monomer comprises sugared analogies.In some this embodiment, replace connecting key combination between sugar or sugar-nucleoside with analogies, keep the nuclear base with selected target hybridization.The representative example of sugar analogies includes but not limited to cyclohexenyl group or morpholino.The representative example that connects the analogies of key combination between sugar-nucleoside includes but not limited to that not having chirality by no electric charge connects peptide nucleic acid(PNA) (PNA) and the morpholino group that key connects.In some cases, replace the nuclear base with analogies.Representational nuclear base analogies are well known in the art, include but not limited to three Huan phenoxazine analog and universal base (28:2911-14 is attached to herein by reference for Berger et al., Nuc Acid Res.2000).The method of synthetic sugared analogies, nucleoside analogies and nuclear base analogies is well known to a person skilled in the art.

3. Dan Julian key

Described herein is with monomer (including but not limited to modified and not modified nucleoside and nucleotide) thereby the linking group of the formation oligomeric compounds that links together.According to whether having phosphorus atoms, linking group mainly is divided into two classes.The representational phosphorous key that connects includes but not limited to di-phosphate ester (P=O), phosphotriester, methyl phosphonate, phosphoramidate and thiophosphate (P=S).Representational non-phosphorous linking group includes but not limited to methylene methyl-imino (CH₂-N (CH₃)-O-CH₂-), the sulfur diester (O-C (O)-S-), thionocarbamate (O-C (O) (NH)-S-), siloxanes (O-Si (H) 2-O-) and N, N '-dimethylformamide dimethyl hydrazine (CH₂-N (CH₃)-N (CH₃)-).Oligomeric compounds with non-phosphorus linking group is called oligonucleoside.Connect key than natural phosphodiester, modified company's key can be used to change the nuclease resistance of (normally improving) oligomeric compounds.In certain embodiments, the Lian Jianke with chiral atom is prepared into racemic mixture, is prepared into independent enantiomer.Representational chirality connects key and includes but not limited to that phosphonate ester and thiophosphate prepare phosphorous even key and the non-phosphorous method that connects key is well known to a person skilled in the art.

Oligomeric compounds described herein contains one or more asymmetric centers, therefore produce enantiomer, diastereomer and other stereoisomerism configurations, determine with the absolute stereo chemistry, for for example sugared anomer is (R) or (S), and perhaps waiting for for example aminoacid is (D) or (L).Antisense compounds provided herein comprises all this possible isomers and their racemic form and the pure form of optically-active.

4. oligomeric compounds

In certain embodiments, this paper provides such oligomeric compounds, and it has to can be used for forming and comprises the reactive phosphorus group that for example connects company's key of key between di-phosphate ester and thiophosphate nucleoside.The method of the precursor of preparation and/or purification oligomeric compounds does not constitute the restriction to compositions provided herein and method.Synthetic and purification comprises that the method for the oligomeric compounds of DNA, RNA, oligonucleotide, oligonucleoside and antisense compounds is well known to a person skilled in the art.

In general, oligomeric compounds comprises a plurality of poly-subunits of list that linked together by linking group.The limiting examples of oligomeric compounds comprises primer, probe, antisense compounds, antisense oligonucleotide, external guide sequence (EGS) oligonucleotide, replaces montage (alternatesplicer) and siRNA.Thus, these chemical compounds can be introduced with the form of strand, two strands, annular, branch or hair clip, and can contain the structural detail such as inside or terminal process (bulge) or ring (loop).The oligomerization double chain compound can be that two chains that hybridization forms double chain compound take place, or has that enough self is complementary so that hybridization takes place and form the single chain of double chain compound wholly or in part.

In certain embodiments, the invention provides chimeric oligomeric compounds.In some this embodiment, chimeric oligomeric compounds is a chimeric oligonucleotide.In some this embodiment, chimeric oligonucleotide comprises the nucleotide through different modifying.In certain embodiments, chimeric oligonucleotide is the mixed matrix antisense oligonucleotide.

In general, chimeric oligomeric compounds can have so modified nucleoside, and they can be in position separately, perhaps can determine that clustering together in the zone of specific die body in meeting.Any combination of modifying and/or simulating group can constitute chimeric oligomeric compounds as herein described.

In certain embodiments, chimeric oligomeric compounds comprises at least one modified zone usually, and this modification can cause to the increase of the raising of the resistance of nuclease degradation, cellular uptake and/or to the raising of the binding affinity of target nucleic acids.In certain embodiments, the substrate of the enzyme that can cut RNA:DNA or RNA:RNA crossbred can be served as in the other zone of oligomeric compounds.For instance, this cell endonuclease of RNA enzyme H can cut the RNA chain of RNA:DNA duplex.Therefore, the activation of RNA enzyme H can cause the cutting of RNA target, thereby greatly strengthens the efficient to the inhibition of gene expression.Therefore, when using chimera, than for example with the thiophosphate deoxy-oligonucleotide of identical target area hybridization, also often can obtain suitable result with short oligomeric compounds.The available gel electrophoresis of the cutting of RNA target carries out conventional sense, if necessary, detects with relevant nucleic acid hybridization technique well known in the art.

In certain embodiments, chimeric oligomeric compounds is gapmer.In certain embodiments, chimeric chemical compound is short antisense compounds.In certain embodiments, lacking antisense compounds is gapmer.In some this embodiment, the mixed matrix antisense oligomers has one type internucleotide linkage in one or two pterion, have dissimilar internucleotide linkages at spacer.In some this embodiment, the mixed matrix antisense oligonucleotide has di-phosphate ester in the pterion and connects key, has thiophosphate at spacer and connects key.At some wherein in the embodiment different of the internucleotide linkage in certain pterion, that the internucleotide linkage of this pterion and spacer bridge joint is identical with internucleotide linkage in the pterion with the internucleotide linkage in the spacer.At some wherein in the embodiment different of the internucleotide linkage in certain pterion, that the internucleotide linkage of this pterion and spacer bridge joint is identical with internucleotide linkage in the spacer with the internucleotide linkage in the spacer.

C. some lacks antisense compounds

Disclosed herein is length 8-16, preferred 9-15, more preferably 9-14, the more preferably short antisense compounds of 10-14 nucleotide.In certain embodiments, the length of lacking antisense compounds is 9-14 nucleotide.In certain embodiments, the length of lacking antisense compounds is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, lack antisense compounds and comprise one or more chemical modifications.In some this embodiment, short antisense compounds comprises at least one modified nucleotide.In certain embodiments, lack antisense compounds and comprise two or more at least modified nucleotide.In certain embodiments, lack antisense compounds and comprise at least one modified internucleotide linkage.In certain embodiments, lacking antisense compounds is the mixed matrix oligonucleotide.In certain embodiments, lacking antisense compounds is chimeric oligonucleotide.In certain embodiments, lack antisense oligonucleotide through evenly modifying.In certain embodiments, lacking antisense oligonucleotide comprises and independently is chosen in each nuclear base and connects the modification of key at each.

In certain embodiments, lacking antisense compounds is short gapmer.In some this embodiment, short gamper comprises at least one high-affinity and modifies in one or more pterions of chemical compound.In certain embodiments, lack antisense compounds and in each pterion, comprise 1-3 high-affinity modification.In certain embodiments, the high-affinity of lacking antisense compounds is modified and to be made the target affinity be similar to and even greater than the target affinity of longer antisense compounds.In certain embodiments, the nucleotide of modifying through high-affinity is through sugar-modified nucleotide.This nucleotide that comprises abutment between those 4 ' and 2 ' positions that comprises through sugar-modified nucleotide at sugar.Sugar-modified BNA and other 2 '-modification as the 2 '-MOE of including but not limited to of exemplary high-affinity.In alternative embodiment of the present invention, it is not 2 '-O-(CH that high-affinity is modified₂)_nThe sugar-modified nucleotide of H (n=1-6).In other alternative embodiment, the nucleotide of modifying through high-affinity is not 2 '-OCH₃Or 2 '-OCH₂CH₂OCH₃Nucleotide.In certain embodiments, the nucleotide of modifying through high-affinity is given the Δ T of at least 1, at least 1.5, at least 2, at least 2.5, at least 3.0, at least 3.5 or at least 4.0 degree/nucleotide_mSome high-affinity nucleotide modifications can increase toxicity known in the art.As shown here, short antisense compounds with high-affinity modification of finite population (2-6 usually) demonstrates few the increasing even of toxicity to be increased, but keeping or improved affinity target RNA, also reduce significantly simultaneously the expression of RNA target.Short antisense compounds of the present invention can be chosen wantonly and comprise conjugated group, for example cholesterol or C₁₆

1. some pterion

In certain embodiments, lack antisense compounds and comprise 5 ' pterion and/or 3 ' pterion.In this embodiment, the feature in the feature in 3 ' pterion and 5 ' pterion is independent the selection.Therefore, in this embodiment, the monomer number (length) in the monomer number in the 5 ' pterion and the 3 ' pterion can be identical or can be different; Modification in the 5 ' pterion (if any) can be identical with the modification (if any) in the 3 ' pterion, and perhaps this modification (if any) can be different; Dan Julian key in Dan Julian key in the 5 ' pterion and the 3 ' pterion can be identical or can be different.

In certain embodiments, the pterion comprises one, two or three monomers (being that length is 1,2 or 3 monomer).In certain embodiments, the monomer in pterion is modified.In some this embodiment, the monomer in pterion is modified to improve the affinity of antisense compounds to its target nucleic acids.In certain embodiments, the monomer in pterion is nucleoside or nucleotide.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion (nucleoside or nucleotide) is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the Dan Julian key in the pterion is natural internucleotide linkage.In certain embodiments, the Dan Julian key in the pterion is between the nucleotide of non-natural or connects key between nucleoside.In certain embodiments, the Dan Julian key in the pterion more can be resisted one or more nucleases than natural internucleotide linkage.In some this embodiment, the Dan Julian key in the pterion is that thiophosphate connects key (P=S).Wherein have in the embodiment of a Dan Julian key of surpassing in the pterion at some, each Dan Julian key is identical mutually.Wherein have in the embodiment of a Dan Julian key of surpassing in the pterion at some, each Dan Julian key is different mutually.

Those of ordinary skills will appreciate that, can with feature discussed above and modify with any be combined into to exercise be used for preparing the pterion.The limiting examples that following table provides has shown how to prepare the pterion by the Dan Julian key of selecting certain monomer number, list to gather in the middle of modification (if any) and the pterion.

Length	Monomer type/modification	Dan Julian key in the middle of the pterion
			1	2’MOE	Do not have
1	BNA	Do not have
			1	Methylene oxygen base BNA	Do not have
1	ENA	Do not have
			2	2’MOE	P＝S
2	BNA	P＝S
			2	Methylene oxygen base BNA	P＝S
2	ENA	P＝S
			2	2’MOE	P＝O
2	BNA	P＝O
			2	Methylene oxygen base BNA	P＝O
2	ENA	P＝O
			3	2’MOE	P＝S
3	BNA	P＝S

3	Methylene oxygen base BNA	P＝S
			3	ENA	P＝S
3	2’MOE	P＝O
			3	BNA	P＝O
3	Methylene oxygen base BNA	P＝O
			3	ENA	P＝O

Wherein comprise in two, three or four the monomeric embodiments in the pterion at some, these two, three or four monomers all comprise identical modification (if any).Wherein comprise in two, three or four the monomeric embodiments in the pterion at some, in these two, three or four nuclear bases one or more different modifications of one or more and one or more all the other monomeric one or more modifications that comprise are arranged.

2. some is at interval

In certain embodiments, lacking antisense compounds comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the Dan Julian key in the interval is natural internucleotide linkage.In certain embodiments, the Dan Julian key in the interval is company's key of non-natural.In some this embodiment, the Dan Julian key in more can be resisted one or more nucleases than natural internucleotide linkage at interval.In some this embodiment, the Dan Julian key in is that thiophosphate connects key (P=S) at interval.In certain embodiments, each the Dan Julian key in the interval is all identical mutually.In certain embodiments, each the Dan Julian key in the interval is not identical all.

Those of ordinary skills will appreciate that, can with feature discussed above and modify with any be combined into to exercise be used for preparation at interval.The limiting examples that following table provides has shown how to prepare at interval by the Dan Julian key of selecting certain monomer number, list to gather in the middle of modification (if any) and the interval region.

Length	Monomer type/modification	Dan Julian key at interval
			5	DNA	P＝S
6	DNA	P＝S
			7	DNA	P＝S
8	DNA	P＝S
			9	DNA	P＝S
10	DNA	P＝S
			11	DNA	P＝S
12	DNA	P＝S
			13	DNA	P＝S
14	DNA	P＝S
			6	DNA	P＝O
7	DNA	P＝O
			8	DNA	P＝O
9	DNA	P＝O
			10	DNA	P＝O
11	DNA	P＝O
			12	DNA	P＝O
8	RNA	P＝S
			9	RNA	P＝S
10	RNA	P＝S
			11	RNA	P＝S
12	RNA	P＝S

Some interband every the antisense oligomeric compounds

Those of ordinary skills will appreciate that, can be selected pterion discussed above and interval, make up with multiple compound mode then, with produce interband every oligomeric compounds, include but not limited to interband every the antisense oligomeric compounds and interband every antisense oligonucleotide.The feature in 5 ' pterion and 3 ' pterion (length, modification, Lian Jian) can be selected independently of each other.Feature at interval comprises at least one and the relative modification difference of feature in 5 ' pterion and at least one and the relative modification difference in 3 ' pterion (being that at least one modification difference must be arranged between each adjacent area, so that these adjacent areas are distinguished mutually).Each feature at interval can be selected in addition independently.

In certain embodiments, the Dan Julian key in the middle of the pterion is identical with central at interval Dan Julian key.In certain embodiments, the Dan Julian key in the middle of the pterion is different with central at interval Dan Julian key.In some this embodiment, that the Dan Julian key of pterion and bridge joint at interval is identical with Dan Julian key in the pterion.In certain embodiments, with the pterion with at interval the Dan Julian key of bridge joint is identical with Dan Julian key in the interval.In certain embodiments, lack antisense compounds and have the same company's key from the beginning to the end.In some this embodiment, all keys all are thiophosphate (P=S) Lian Jian.

Those of ordinary skills will appreciate that, can with 3 ' pterion discussed above, 5 ' pterion, interval and connect key with any be combined into to exercise be used for preparing gapmer.The limiting examples that following table provides how to have shown by select a certain 5 ' pterion, at interval, 3 ' pterion and some will be at interval and company's key of each pterion bridge joint prepare gapmer.

In certain embodiments, oligomeric compounds disclosed herein can comprise about 8 to about 16, preferred 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer) to about 16 connections.Those of ordinary skills will appreciate that this comprises the antisense compounds of 8,9,10,11,12,13,14,15 or 16 nuclear bases.In certain embodiments, oligomeric compounds is an antisense compounds.

In certain embodiments, the length of lacking antisense compounds is 8 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 9 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 10 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 11 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 12 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 13 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 14 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 15 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 16 nucleotide.

In certain embodiments, the length of lacking antisense compounds is 8 monomers.In certain embodiments, the length of lacking antisense compounds is 9 monomers.In certain embodiments, the length of lacking antisense compounds is 10 monomers.In certain embodiments, the length of lacking antisense compounds is 11 monomers.In certain embodiments, the length of lacking antisense compounds is 12 monomers.In certain embodiments, the length of lacking antisense compounds is 13 monomers.In certain embodiments, the length of lacking antisense compounds is 14 monomers.In certain embodiments, the length of lacking antisense compounds is 15 monomers.In certain embodiments, the length of lacking antisense compounds is 16 monomers.In certain embodiments, lack antisense compounds and comprise 9-15 monomer.In certain embodiments, lack antisense compounds and comprise 10-15 monomer.In certain embodiments, lack antisense compounds and comprise 12-14 monomer.In certain embodiments, lack antisense compounds and comprise 12-14 nucleotide or nucleoside.

Those skilled in the art have learnt that behind the illustrational short antisense compounds of this paper institute, it goes without doing, and too many experiment just can be determined more short antisense compounds.

In certain embodiments, lacking antisense compounds comprises its one or both sides and is connected to the interval that surpasses a pterion.Therefore, in certain embodiments, short antisense compounds comprises two or more 5 ' pterions and two or more 3 ' pterion.In certain embodiments, lack antisense compounds and comprise one 5 ' pterion and two or more 3 ' pterion.In certain embodiments, lack antisense compounds and comprise one 3 ' pterion and two or more 5 ' pterion.Some this embodiment comprises for example with lower area: the 1 ' pterion-abutment-the 25 ' pterion-abutment-at interval-and abutment-the 23 ' pterion-abutment-the 1 ' pterion.In this embodiment, each zone is adjacent the zone and compares and have at least one and modify difference.Therefore pterion and the 1,, in this embodiment, the 25 ' pterion and the 23 ' pterion with compare at interval with the 1 ' ' pterion compares, and independently comprises one or more modification differences separately.In this embodiment, the 1 ' modification and the 1 in pterion ' one of the modification in pterion or both can be identical or different with modification (if any) at interval.

4. some conjugated group

In one aspect, oligomeric compounds is to modify by the covalently bound of one or more conjugated groups.In general, conjugated group can be modified one or more characteristics of connected oligomeric compounds, includes but not limited to pharmacodynamics, pharmacodynamics, combination, absorption, cell distribution, cellular uptake, electric charge and removing.Conjugated group is directly or by optional coupling part or linking group to be connected with parent compound such as oligomeric compounds in the conventional use of chemical field.The preferred list of conjugated group includes but not limited to intercalator (intercalator), reporter molecule, polyamine, polyamide, Polyethylene Glycol, thioether, polyethers, cholesterol, sulfo-cholesterol, cholic acid part, folic acid, lipid, phospholipid, biotin, azophenlyene, phenanthridines, anthraquinone, diamantane (obsolete), acridine, fluorescein, rhodamine, coumarin and dyestuff.

Preferably be suitable for conjugated group of the present invention and comprise lipid part such as cholesterol moiety (Letsinger et al., Proc.Natl.Acad.Sci.USA, 1989,86,6553); Cholic acid (Manoharan et al., Bioorg.Med.Chem.Lett., 1994,4,1053); Thioether, for example hexyl-S-trityl mercaptan (Manoharan et al., Ann.N.Y.Acad.Sci., 1992,660,306; Manoharan et al., Bioorg.Med.Chem.Let., 1993,3,2765); Sulfo-cholesterol (Oberhauser et al., Nucl.Acids Res., 1992,20,533); Aliphatic chain, for example dodecanediol or or undecyl residue (Saison-Behmoaras et al., EMBO J., 1991,10,111; Kabanov et al., FEBSLett., 1990,259,327; Svinarchuk et al., Biochimie, 1993,75,49); Phospholipid, for example two-cetyl-rac-glycerol or triethyl ammonium-1,2-two-O-cetyl-rac-glycerol-3-H-phosphate ester (Manoharan et al., Tetrahedron Lett., 1995,36,3651; Sheaet al., Nucl.Acids Res., 1990,18,3777); Polyamine or polyglycol chain (Manoharan et al., Nucleosides ﹠amp; Nucleotides, 1995,14,969); Adamantane acetic acid (Manoharan et al., Tetrahedron Lett., 1995,36,3651); Palmityl part (Mishra et al., Biochim.Biophys.Acta, 1995,1264,229); Perhaps 18-amine. or hexyl amino-carbonyl-oxycholesterin part (Crooke et al., J.Pharmacol.Exp.Ther., 1996,277,923).

Linking group or difunctionality coupling part (for example well known in the art those) are suitable for chemical compound provided herein.Linking group can be used for chemical functional group, conjugated group, reporter group and other groups and parent compound for example are connected in the selectivity site in the oligomeric compounds.In general, the difunctionality coupling part comprises the hydrocarbyl portion with two functional groups.One of them functional group is through selecting with binding purpose parent molecule or chemical compound, and another is through selecting with in conjunction with any selected group basically, as chemical functional group or conjugated group.In some embodiments, joint (linker) comprises unitary chain structure of repetitive such as ethylene glycol or aminoacid or oligomer.Routine is used for the example of the functional group of difunctionality coupling part, includes but not limited to the nucleophile that is used for the electrophile of nucleophilic group reaction and is used for reacting with electrophilic group.In some embodiments, the difunctionality coupling part comprises amino, hydroxyl, carboxylic acid, mercaptan, unsaturated bond (for example two keys or triple bond) etc.Some limiting examples of difunctionality coupling part comprise 8-amino-3,6-two oxa-s sad (ADO), 4-(N-maleimide ylmethyl) cyclohexane extraction-1-formic acid succinimide ester (SMCC) and 6-aminocaprolc acid (AHEX or AHA).Other linking group includes but not limited to substituted C₁-C₁₀Alkyl, substituted or unsubstituted C₂-C₁₀Substituted or the unsubstituted C of thiazolinyl₂-C₁₀Alkynyl, wherein the non-limiting list of preferred substituted comprises hydroxyl, amino, alkoxyl, carboxyl, benzyl, phenyl, nitro, mercaptan, thio alkoxy, halogen, alkyl, aryl, thiazolinyl and alkynyl.

5. synthetic, purification and analysis

Modified and the not modified nucleoside and the oligomerization of nucleotide, can be according to program (the Protocols for Oligonucleotides and Analogs that in the document DNA is reported, Ed.Agrawal (1993), Humana Press) and/or to the program (Scaringe of RNA report, Methods (2001), 23,206-217.Gait et al., Applications of Chemicallysynthesized RNA in RNA:Protein Interactions, Ed.Smith (1998), 1-36.Gallo et al., Tetrahedron (2001), 57,5707-5713) carry out.

Oligomeric compounds provided herein can be prepared expediently and routinely by the known technology in the solid phase synthesis.Have several manufacturers to sell and carry out this synthetic equipment, for example comprise Applied Biosystems (Foster City, CA).In addition or or, can adopt this synthetic any other means of carrying out well known in the art.Use similar techniques to prepare oligonucleotide such as thiophosphate and alkyl derivative, this is well-known.The present invention is not subjected to the restriction of antisense compounds synthetic method.

The method of purification and analysis oligomeric compounds is well known to a person skilled in the art.Analytical method comprises capillary electrophoresis (CE) and electron spray-mass spectrum.This synthesizing with analytical method can carry out in porous plate.Method of the present invention is not subjected to the restriction of oligomer purification process.

D. antisense mechanism

Antisense mechanism (mechanism) is the mechanism that all that relates to the hybridization of certain chemical compound and target nucleic acids, wherein Za Jiao consequence or effect are that target degraded or target occupy, and are accompanied by to relate to for example to transcribe or the stopping of cell mechanism (the cellular machinery) of montage.

One type the antisense mechanism that relates to the target degraded comprises RNA enzyme H.RNA enzyme H is a kind of cell endonuclease that can cut the RNA chain of RNA:DNA duplex.Known in this field, the strand antisense compounds of " DNA sample " can cause the RNA enzyme H activity in the mammalian cell.Therefore the activation of RNA enzyme H causes the cutting of RNA target, thereby greatly strengthens the efficient of the oligonucleotide mediated inhibition to gene expression of DNA sample.

In certain embodiments, through the antisense compounds of chemical modification to the affinity of target RNA than non-modified DNA height.In some this embodiment, this higher affinity causes the raising of usefulness, the feasible this chemical compound that can give than low dosage, and the toxicity probability descends, and therapeutic index is improved and whole medical expense descends.

Present disclosure specification proves, to be incorporated in the antisense compounds through the high-affinity nucleotide and the nucleoside of chemical modification, can make and to design length 8-16 the short antisense compounds of examining base, target RNA and/or target proteins matter that this weak point antisense compounds can be used for reducing cell, tissue and animal (including but not limited to the people) with the usefulness and the improved therapeutic index of raising.Therefore, in certain embodiments, the present invention is to provide so short antisense compounds, it comprises the high-affinity nucleotide modification that can be used for reducing target RNA in the body.Some this short antisense compounds also can be effective under than the lower dosage of the antisense compounds of describing before, and this makes and can reduce toxicity and treatment cost.In addition, some short antisense compounds has bigger oral administration probability.

Be to satisfy the demand to more potent antisense compounds, this paper provides the short antisense compounds comparing activity in vivo with longer chemical compound and improve (length 8-16, preferably 9-15, more preferably 9-14, more preferably 10-14 nucleotide).Some short antisense compounds is the gapmer chemical compound that comprises in 3 ' and 5 ' terminal (pterion) of chemical compound through the nucleotide of high-affinity chemical modification.In certain embodiments, although the adding of the nucleotide of modifying through high-affinity can make that antisense compounds length is shorter, target RNA in its predetermined body there are activity and specificity.What this paper imagined is so short antisense compounds, and wherein each pterion independently comprises 1-3 the nucleotide of modifying through high-affinity.In certain embodiments, this high-affinity modification is sugar-modified.The nucleotide of modifying through high-affinity includes but not limited to BNA and other nucleotide through 2 '-modification, as 2 '-MOE nucleotide.Also imagination is the short antisense compounds with at least one modified internucleotide linkage (for example thiophosphate internucleotide linkage).In certain embodiments, short antisense compounds of the present invention can have between all thiophosphate nucleoside and connects key.The optional conjugated group that comprises of short antisense compounds.As shown here, short antisense compounds is stronger to the affinity of DNA than them to the affinity of target RNA, and shows that by the minimizing of target mRNA and the minimizing of multiple disease indication they are more potent significantly in vivo.

This paper uses this point, promptly participate in to regulate glucose metabolism or removing, lipid metabolism, cholesterol but thanks or the RNA of insulin metabolism, is the RNA in any biological approach that participates in these processes of adjusting.This RNA is well known in the art.The example of target gene includes but not limited to that ApoB-100 (also claims APOB; Ag (x) antigen; ApoB-48; Apolipoprotein B; Apolipoprotein B-100; Apo B-48) and GCGR (also claim glucagon receptor; GR), CRP, DGAT2, GCCR, PCSK9, PTEN, PTP1B, SGLT2 and SOD1.

1. the adjusting of target expression

In certain embodiments, identify target and design the antisense oligonucleotide that to regulate this target or its expression.In certain embodiments, the oligomeric compounds of design targeting target nucleic acids molecule, this may be a rapid process of multistep.Usually, the target proteins matter of this process from identifying that its activity is to be regulated identifies that then its expression can produce the nucleic acid of this target proteins matter.In certain embodiments, the design of antisense compounds cause obtaining can with by the antisense compounds of the making nucleic acid molecular hybridization of targeting.In certain embodiments, this antisense compounds is antisense oligonucleotide or antisense oligonucleoside.In certain embodiments, antisense compounds and target nucleic acids are complementary mutually.In some this embodiment, antisense compounds and target nucleic acids are perfect complementary.In certain embodiments, antisense compounds comprises a mispairing.In certain embodiments, antisense compounds comprises two mispairing.In certain embodiments, antisense compounds comprises three or more mispairing.

The adjusting of the expression of target nucleic acids can realize by the nucleic acid function that changes any number.In certain embodiments, the function of RNA to be regulated includes but not limited to easy bit function, and the transposition function includes but not limited to that RNA translocates to the position of protein translation, RNA and translocates in the middle of the cell and carry out the position that the position of the synthetic and protein of RNA from the translation of this RNA is far apart.The RNA machining functions that can be regulated includes but not limited to that the montage (causing producing one or more RNA kinds (RNA species)) of this RNA, the 3 ' maturation that adds medicated cap, this RNA and the catalytic activity that relates to this RNA or complex of this RNA form (it can participate among (engaged in) this RNA or by this RNA and promote).The adjusting of expressing can cause the level of one or more nucleic acid species (nucleic acid species) to improve or the level of one or more nucleic acid species reduces, and described raising or reduction are interim or are in clean steady-state level (by netsteady state level).Therefore, in one embodiment, the adjusting of expression can mean the raising or the reduction of target RNA or protein level.In another embodiment, the adjusting of expression can mean the raising or the reduction of one or more RNA montage products, the perhaps variation of the ratio of two or more montage products.

In certain embodiments, the expression of target gene be with comprise about 8 to about 16, preferred 9-15, more preferably 9-14, more preferably the oligomeric compounds of 10-14 monomer (promptly about 8 monomer to about 16 connections) is regulated.Those of ordinary skills will appreciate that this comprises the method that the antisense compounds of one or more 8,9,10,11,12,13,14,15 or 16 nuclear of use bases is regulated the expression of target gene.

In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 8 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 9 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 8 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 10 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 10 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 11 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 12 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 13 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 14 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 15 nuclear bases.In certain embodiments, the method for adjusting target gene comprises that using length is the short antisense compounds of 16 nuclear bases.

In certain embodiments, the method for the expression of adjusting target gene comprises that use comprises 9-15 monomeric short antisense compounds.In certain embodiments, the method for the expression of adjusting target gene comprises that use comprises 10-15 monomeric short antisense compounds.In certain embodiments, the method for the expression of adjusting target gene comprises that use comprises 12-14 monomeric short antisense compounds.In certain embodiments, the method for the expression of adjusting target gene comprises that use comprises the short antisense compounds of 12 or 14 nucleotide or nucleoside.

2. hybridization

In certain embodiments, antisense compounds as enough complementary degree when avoiding under the bonded condition of needs specificity antisense compounds and non-target nucleic acids sequence generation non-specific binding, then it is with regard to the energy specific hybrid, described condition measure in vivo or medical treatment treatment situation under be physiological condition, be the condition of carrying out this mensuration in the situation of external test.

" stringent hybridization condition " used herein or " stringent condition " refer to that antisense compounds can hybridize with its target sequence but the condition few with the hybridization of other sequences.Stringent condition is sequence-dependent, can be different in different situations." stringent condition " of the hybridization of antisense compounds and target nucleic acids is by the characteristic of antisense compounds and forms and reach the algoscopy of studying them according to this and determine.

3. complementary

This area recognizes that mixing that the nucleotide affinity is modified can make than not modified chemical compound the more mispairing of more number is arranged.Similarly, comparable other oligonucleotide sequences of some few nucleotide sequence more can be tolerated mispairing.Those of ordinary skills can be for example by measuring melting temperature (T_m), determine the suitable mispairing number between each oligonucleotide or between certain oligonucleotide and the target nucleic acids.T_mOr Δ T_mCan go out by the technique computes that those of ordinary skills know.For example, and Freier etc. (Nucleic Acids Research, 1997,25,22:4429-4443) technology of Miao Shuing can allow those skilled in the art assess the ability of nucleotide modification aspect the melting temperature that improves the RNA:DNA duplex.

4. homogeneity

Antisense compounds or its part can have definite (defined) homogeneity percentage rate with certain SEQ ID NO or the chemical compound with specific Isis number.As used herein, promptly certain sequence is when it and sequence disclosed herein have identical nuclear base pairing ability, and then the sequence of it and the disclosure has homogeneity.For example, wherein contain uracil and can be considered to homogeneity, because uracil and thymidine can both match with adenine with the RNA of the thymidine (thymidine) in the open sequence that replaces chemical compound described herein.This homogeneity can be on the whole length of oligomeric compounds, perhaps in the part of antisense compounds (for example, nuclear base 1-20 and certain 20-mer of certain 27-mer can be compared, to determine the homogeneity percentage rate of this oligomeric compounds and this SEQ ID NO).Those skilled in the art recognize that certain antisense compounds does not need to have with the same sequence of those chemical compounds described herein and just can play and the similar function of antisense compounds described herein.The present invention also provides the shortening version (version) of antisense compounds teaching herein, the nonidentity version of antisense compounds perhaps teaching herein.The nonidentity version is that wherein each base does not have active those antisense compounds of the pairing identical with antisense compounds disclosed herein.Each base is because shorter or have at least one abasic site (abasic site) and do not have identical pairing activity.Perhaps, the nonidentity version can comprise that at least one is had the base (for example G can be replaced by C, A or T) that the active different bases of different pairings have replaced.The homogeneity percentage rate is to calculate according to the base number that has with the same base pairing corresponding to the SEQ ID NO that is compared or antisense compounds.The nonidentity base can be adjacent each other, perhaps can be distributed in oligonucleotide everywhere, and perhaps both of these case has.

For example, have certain 16-mer of the sequence identical, 80% homogeneity is arranged with this 20-mer with the nuclear base 2-17 of certain 20-mer.Perhaps, contain four nuclear bases with different certain 20-mer of this 20-mer, also 80% homogeneity is arranged with this 20-mer.Have certain 14-mer of the sequence identical, 78% homogeneity is arranged with this 18-mer with the nuclear base 1-14 of certain 18-mer.This calculating is fully in those skilled in the art's limit of power.

The homogeneity percentage rate is based on there is ratio in nuclear base in the original series in modified sequence percent.Therefore, the 30 nuclear base antisense compounds of sufficient sequence that comprise the complementary series (complement) of the 20 nuclear active target sections of base (target segment), can have the part that 100% homogeneity is arranged with the complementary series of the active target section of this 20 nuclear base, also comprise 10 other nuclear base portions simultaneously.In the situation of this description, the complementary series of active target section can constitute single part.In preferred embodiments, at least a portion of the complementary series of the active target section that provides of oligonucleotide provided herein and this paper has at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% homogeneity.

E. target nucleic acids, zone and section

In certain embodiments, can design short antisense compounds and come any target nucleic acids of targeting.In certain embodiments, the relevant clinically target of target nucleic acids coding.In this embodiment, can produce clinical benefit to the adjusting of target nucleic acids.Some target nucleic acids includes but not limited to the target nucleic acids that table 1 is listed.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding ApoB.The nucleic acid molecules of coding ApoB includes but not limited to SEQ ID NO:1 and SEQ ID NO:2.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding SGLT2.The nucleic acid molecules of coding SGLT2 includes but not limited to SEQ ID NO:3.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding PCSK9.The nucleic acid molecules of coding PCSK9 includes but not limited to SEQ ID NO:4.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding SOD1.The nucleic acid molecules of coding SOD1 includes but not limited to SEQ ID NO:5.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding CRP.The nucleic acid molecules of coding CRP includes but not limited to SEQ ID NO:3.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding GCCR.The nucleic acid molecules of coding GCCR includes but not limited to SEQ ID NO:7 and SEQ ID NO:8.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding GCGR.The nucleic acid molecules of coding GCGR includes but not limited to SEQ ID NO:9.

In certain embodiments, target nucleic acids is the nucleic acid molecules of encoding D GAT2.The nucleic acid molecules of encoding D GAT2 includes but not limited to SEQ ID NO:10.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding PTP1B.The nucleic acid molecules of coding PTP1B includes but not limited to SEQ ID NO:11 and SEQ ID NO:12.

In certain embodiments, target nucleic acids is the nucleic acid molecules of coding PTEN.The nucleic acid molecules of coding PTEN includes but not limited to SEQ ID NO:14 or SEQ ID NO:15.

Table 1: some target nucleic acids

The targeting process generally includes at least one such target zone, section or site of determining that target nucleic acids is central, and this target zone, section or site can make the antisense interaction occur, and make to obtain required effect.

In certain embodiments, 5 '-least significant end nucleotide in target zone is 5 ' target site of short antisense compounds, and 3 '-least significant end nucleotide in target zone is 3 ' target site of this identical short antisense compounds.In certain embodiments, 5 '-least significant end nucleotide in target zone is 5 ' target site of short antisense compounds, and 3 '-least significant end nucleotide in target zone is 3 ' target site of different short antisense compounds.In certain embodiments, the target zone comprises apart from 5 ' target site or the interior nucleotide sequence of 10,15 or 20 the nucleotide scopes in 3 ' target site.

In certain embodiments, the target zone is the zone of determining (structurallydefined) on the structure of nucleic acid.For example, in some this embodiment, 3 ' UTR, 5 ' UTR, exon, intron, coding region, translation initiation district, translation termination district or other nucleic acid region of determining can be contained in the target zone.

On the target nucleic acids by having one or more active short antisense compounds to the determined position of its targeting, be called " active target section ".In certain embodiments, having one or more active short antisense compounds is target RNA to the target nucleic acids of its targeting.When active target section is when being determined by a plurality of short antisense compounds, these chemical compounds preferably are separated by on target sequence and are no more than about 10 nucleotide, more preferably on target sequence, be separated by and be no more than about 5 nucleotide, also more preferably these short antisense compounds are successive, and most preferably these short antisense compounds are eclipsed.The short antisense compounds of in the middle of the active target section each has great difference on active (for example being determined by suppressing percent).Active short antisense compounds is those the short antisense compounds that can regulate the expression of its target nucleic acids (including but not limited to target RNA).Active short antisense compounds can suppress the expression at least 10% of its target RNA, preferred 20%.In a preferred embodiment, at least about 50%, the short antisense compounds of preferred about 70% the active target section of targeting can regulate the expression at least 40% of its target RNA.Determining the required inhibition level of active short antisense compounds at one more in the embodiment preferred, is to determine according to the result in order to the screening gained of determining active target section.

Suitable target section be active short antisense compounds institute targeting the target zone at least about 8 nuclear base portions.The target section can comprise such DNA or RNA sequence, these sequences comprise 5 ' of a target section from each illustrative (illustrative) target section-terminal at least 8 continuous kernel bases (remaining nuclear base is so continuous one section sequence (consecutive stretch) of same DNA or RNA, this section sequence just in time the upstream of 5 ' of this target section-end begin and proceed to this DNA or RNA comprise about 8 examine bases to about 16 till).The target section is also represented by such DNA or RNA sequence, these sequences comprise 3 ' of a target section from each illustrative target section-terminal at least 8 continuous kernel bases (remaining nuclear base is so continuous one section sequence of same DNA or RNA, this section sequence just in time the downstream of 3 ' of this target section-end begin and proceed to this DNA or RNA comprise about 8 examine bases to about 16 till).Also will be appreciated that, antisense target section can be represented by such DNA or RNA sequence, these sequences comprise at least 8 continuous kernel bases from the interior section of the sequence of certain illustrative target section, and can extend to short antisense compounds along either direction or both direction and comprise about 8 till about 16 nuclear bases.Those skilled in the art rely on the illustrated target section of this paper, and it goes without doing, and too much experiment just can identify more target section.

In case identify one or more targets zone, section or site, just can select the short antisense compounds that target is had enough complementarity, can be enough fully and with enough specificitys the short antisense compounds of hybridization takes place, to produce required effect.

Also can be with the such zone in the short antisense compounds targeting target nuclear base sequence, this zone comprises any continuous kernel base of length 8-16 nuclear base on the target nucleic acids molecule.

Target section length 8-16 nuclear base, that comprise one section sequence (stretch) of at least 8 continuous kernel bases selecting in the middle of the illustrative target section is also thought to be suitable for carrying out targeting.Therefore, lacking antisense compounds also can contain 8-16 that this paper confirms as in the middle of those sections of beginning in certain specific 5 ' target site and examine base.Around these zones 50, preferred 25, more preferably any 8,9,10,11 in 16 the nuclear base scopes (perimeter) or the more preferably section of 12,13,14,15 or 16 continuous kernel bases, also think to be suitable for carrying out targeting.

In another embodiment, " the suitable target section " that this paper confirmed can be applied to screen the short antisense compounds of the other expression that can regulate target nucleic acids." regulator " is such chemical compound, and they can reduce or improve the expression of target nucleic acids, includes the complementary 8 nuclear base portions of at least one and target section.Screening technique comprises such step: make the target section of nucleic acid contact and select one or more energy reductions or improve the candidate modulator of the expression of target nucleic acids with one or more candidate modulator.Can regulate the expression of (for example reduce or improve) target nucleic acids in case prove these one or more candidate modulator, this regulator can be applicable to the further research of function do to target then, perhaps uses, diagnoses and use or the medicine for treatment agent as research according to the present invention.

For all short antisense compounds that this paper discusses, sequence, monomer, single poly-modification and Dan Julian key can independently be selected separately.In certain embodiments, lacking antisense compounds is to describe by die body (motif).In this embodiment, any die body can use with any sequence, no matter whether this sequence and/or die body be clearly open in this article.In certain embodiments, lack antisense compounds and comprise the modification that is unsuitable for describing (for example, the diverse location place in the middle of it comprises several different modifying and/or connects the short antisense compounds of key) by die body.Can be any sequence and mix this combination, no matter whether this sequence is open in this article.Originally the appended sequence table of presenting a paper provides some to be independent of the nucleotide sequence of chemical modification.Though this sequence table is confirmed as each sequence " RNA " or " DNA " as required, actually, these sequences can be modified with any combination of chemical modification and/or die body.

In certain embodiments, lack antisense compounds and comprise the monomer that at least one is modified through high-affinity.In certain embodiments, provide the targeting coding to include but not limited to the short antisense compounds of the nucleic acid molecules of following target: ApoB-100 (also claims APOB; Ag (x) antigen; ApoB-48; Apolipoprotein B; Apolipoprotein B-100; Apo B-48), GCGR (also claims glucagon receptor; GR), CRP, DGAT2, GCCR, PCSK9, PTEN, PTP1B, SGLT2 and SOD1.In some this embodiment, the nucleic acid molecules of this short antisense compounds targeting coding any of these target.

F. some target

In certain embodiments, short antisense compounds can be designed to regulate any target.In certain embodiments, target is relevant clinically.In this embodiment, can produce clinical benefit to the adjusting of target.Some target is preferentially expressed in kidney.Some target is preferentially expressed in liver.Some target is relevant with metabolic disease.Some target is relevant with cardiovascular disease.In certain embodiments, target is selected from ApoB, SGLT2, PCSK9, SOD1, CRP, GCCR, GCGR, DGAT2, PTP1B and PTEN.In certain embodiments, target is selected from ApoB, SGLT2, PCSK9, SOD1, CRP, GCCR, GCGR, DGAT2 and PTP1B.In certain embodiments, target is any albumen outside the SGLT2.

In certain embodiments, lack antisense compounds and demonstrate liver and kidney specific target RNA reduction effect in the body.This specific character makes these short antisense compounds be specially adapted to suppress many target RNA that relate to metabolic disease and cardiovascular disease.Therefore, this paper provides the short antisense compounds by making described kidney or liver organization and the targeting RNA relevant with described disease to contact, and treats the method for cardiovascular disease or metabolic disease.Therefore, also provide the method for improving any indication in multiple metabolic disease or the cardiovascular disease indication with short antisense compounds of the present invention.

1.ApoB

ApoB (also claims Apolipoprotein B-100; ApoB-100; Apo B-48; ApoB-48 and Ag (x) antigen) be a kind of big glycoprotein, play indispensable effect in the assembling of lipid and secretion and in the transhipment and the receptor-mediated picked-up of different classes of lipoprotein with in sending.ApoB can carry out multiple activity, from the absorption and the processing of meals lipid, to the adjusting of circulation lipoprotein levels (Davidson and Shelness, Annu.Rev.Nutr., 2000,20,169-193).This back one behavioral illustrations it is why relevant with the atherosclerosis susceptibility, the concentration height correlation on every side of this susceptibility and the lipoprotein that contains ApoB (Davidson and Shelness, Annu.Rev.Nutr., 2000,20,169-193).ApoB-100 is the main protein component of LDL-C, and it contains this lipoprotein classification and the required domain of ldl receptor interaction.High-caliber LDL-C is the cardiovascular disease risk factor that comprises atherosclerosis.

Definition

" ApoB " is a kind of gene outcome or protein, and its expression will be regulated by lacking antisense compounds.

" ApoB nucleic acid " means the nucleic acid of any coding ApoB.For example, in certain embodiments, ApoB nucleic acid include but not limited to the to encode DNA sequence of ApoB, the RNA sequence of transcribing from the DNA of coding ApoB and the mRNA sequence of coding ApoB.

" ApoB mRNA " means the mRNA of coding ApoB.

ApoB treats indication

In certain embodiments, the invention provides the method for the expression of regulating the ApoB in the individuality, described method comprises the short antisense compounds that gives targeting ApoB nucleic acid.In certain embodiments, the invention provides the individual method of treatment, described method comprises and gives the pharmaceutical composition that one or more comprise the short antisense compounds of targeting ApoB nucleic acid.In certain embodiments, individuality suffers from hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, the heterozygous familial hypercholesterolemia, familial hypercholesterolemia isozygotys, the Combination dyslipidemia, atherosclerosis, develop atherosclerotic risk, coronary heart disease, the coronary disease medical history, the coronary heart disease of early showing effect, one or more coronary heart disease risk factors, type ii diabetes, type ii diabetes companion dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipemia, the high fatty acid mass formed by blood stasis, fatty degeneration of liver, non-alcoholic stellato-hepatitis or non-alcohol fatty liver.

The guilding principle that lipid reduces therapy by u.s. national cholesterol education program (NCEP) adult treatment expert scheme III (ATPIII) in calendar year 2001 set up and upgraded in 2004 (Grundyet al., Circulation, 2004,110,227-239).This guilding principle comprises acquisition complete lipoprotein general picture (profile) after fasting in 9-12 hour usually, to determine LDL-C, T-CHOL and HDL-C level.According to the guilding principle of up-to-date formulation, 130-159mg/dL, 160-189mg/dL and think critical height, height and high level respectively more than or equal to the LDL-C level of 190mg/dL.200-239mg/dL and think critical height and high level respectively more than or equal to the total cholesterol level of 240mg/dL.HDL-C level less than 40mg/dL is thought low-level.

In certain embodiments, this individuality has been confirmed as carrying out lipid reduction therapy.In some this embodiment, according to upgrading (Grundyet al. in calendar year 2001 foundation and in 2004 by u.s. national cholesterol education program (NCEP) adult treatment expert scheme III (ATPIII), Circulation, 2004,110, guilding principle 227-239), this individuality have been confirmed as carrying out lipid and have reduced therapy.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 190mg/dL.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 160mg/dL.In some this embodiment, its LDL-C of this individuality that need carry out lipid reduction therapy is higher than 130mg/dL.In some this embodiment, its LDL-C of this individuality that need carry out lipid reduction therapy is higher than 100mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 160mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 130mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 100mg/dL.In some this embodiment, this individuality should keep LDL-C to be lower than 70mg/dL.

In certain embodiments, the invention provides the method that reduces the ApoB in the individuality.In certain embodiments, the invention provides the method that contains the ApoB lipoprotein that reduces in the individuality.In certain embodiments, the invention provides the method that reduces the LDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the VLDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the IDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the non-IDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the Lp (a) in the individuality.In certain embodiments, the invention provides the method that reduces the serum triglycerides in the individuality.In certain embodiments, the invention provides the method that reduces the liver tg in the individuality.In certain embodiments, the invention provides the method that reduces the Ox-LDL-C in the individuality.In certain embodiments, the invention provides the particulate method of little LDL that reduces in the individuality.In certain embodiments, the invention provides the particulate method of little VLDL that reduces in the individuality.In certain embodiments, the invention provides the method that reduces the phospholipid in the individuality.In certain embodiments, the invention provides the method that reduces the oxidized phospholipids in the individuality.

In certain embodiments, the invention provides the method that reduces the Ox-LDL-C concentration among the experimenter.In some this embodiment, the reduction of ApoB, LDL-C, VLDL-C, IDL-C, T-CHOL, non-HDL-C, Lp (a), triglyceride or Ox-LDL-C independently is selected from least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% and at least 100%.In some this embodiment, the reduction of ApoB, LDL-C, VLDL-C, IDL-C, T-CHOL, non-HDL-C, Lp (a), triglyceride or Ox-LDL-C independently is selected from least 20%, at least 30%, at least 40%, at least 50%, at least 60% and at least 70%.In some this embodiment, the reduction of ApoB, LDL-C, VLDL-C, IDL-C, T-CHOL, non-HDL-C, Lp (a), triglyceride or Ox-LDL-C independently is selected from least 40%, at least 50%, at least 60% and at least 70%.

In certain embodiments, the invention provides the method that improves the HDL-C concentration among the experimenter.

In certain embodiments, method provided by the invention does not reduce HDL-C.In certain embodiments, method provided by the invention does not cause the accumulation of lipid in liver.In certain embodiments, method provided by the invention does not cause fatty degeneration of liver.

In certain embodiments, can reduce again and the method for the treatment of relevant side effect when the invention provides ApoB concentration in reducing the experimenter.In some this embodiment, side effect is a liver toxicity.In some this embodiment, side effect is an abnormal liver function.In some this embodiment, side effect is that alanine aminotransferase (ALT) raises.In some this embodiment, side effect is that aspartate aminotransferase (AST) raises.

In certain embodiments, the invention provides the method that reduces the ApoB concentration among the experimenter not have to reach target LDL-C level because of lipid reduction therapy.In some this embodiment, the short antisense compounds of targeting ApoB nucleic acid is that the unique lipid that gives the experimenter reduces medicament.In some this embodiment, experimenter's lipid that acceptance (complywith) recommended of not following the doctor's advice as yet reduces therapy.In some this embodiment, Pharmaceutical composition of the present invention reduces therapy with in addition different lipid and gives altogether together.In some this embodiment, it is the LDL-Apheresis that other lipid reduces therapy.In some this embodiment, it is inhibin that other lipid reduces therapy.In some this embodiment, it is according to Ezetimibe that other lipid reduces therapy.

In certain embodiments, the invention provides the method for the ApoB concentration among the experimenter that reduction do not tolerate inhibin.In some this embodiment, because given inhibin, experimenter's creatine kinase concentration improves.In some this embodiment, because given inhibin, experimenter's abnormal liver function.In some this embodiment, because given inhibin, experimenter's myalgia.In some this embodiment, because given inhibin, the side effect of experimenter's center of origin nervous system.In certain embodiments, experimenter's inhibin of accepting to be recommended of not following the doctor's advice as yet gives.

In certain embodiments, the invention provides the method that reduces the liver tg among the experimenter.In some this embodiment, experimenter's liver tg raises.In some this embodiment, the experimenter suffers from fat hepatitis.In some this embodiment, the experimenter suffers from steatosis.In some this embodiment, the liver tg level is to measure by nuclear magnetic resonance.

In certain embodiments, the invention provides the method that reduces the coronary heart disease risk among the experimenter.In certain embodiments, the invention provides the method for the atherosclerotic progress among the experimenter that slows down.In some this embodiment, the invention provides the method that stops the atherosclerotic progress among the experimenter.In some this embodiment, the invention provides the size of the atherosclerotic plaque that reduces among the experimenter and/or the method for popular (prevalence).In certain embodiments, the method that is provided can reduce the experimenter and develop atherosclerotic risk.

In certain embodiments, the method that is provided can be improved the cardiovascular consequence among the experimenter.In some this embodiment, the improvement of cardiovascular consequence is that the risk of development coronary heart disease reduces.In some this embodiment, the improvement of cardiovascular consequence is the reduction of the incidence rate of one or more main cardiovascular events, and described incident includes but not limited to death, myocardial infarction, infraction, apoplexy, cardiogenic shock, pulmonary edema, cardiac arrest and atrial arrhythmia again.In some this embodiment, the improvement of cardiovascular consequence is confirmed by the improvement of carotid intimal medial thickness.In some this embodiment, the improvement of carotid intimal medial thickness is the reduction of thickness.In some this embodiment, the improvement of carotid intimal medial thickness is preventing of media thickness increase.

In certain embodiments, the pharmaceutical composition that comprises the short antisense compounds of targeting ApoB nucleic acid is for using in treatment.In certain embodiments, described treatment is the reduction of LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp (a), serum triglycerides, liver tg, Ox-LDL-C, little LDL granule, little VLDL, phospholipid or oxidized phospholipids in the individuality.In certain embodiments, described treatment is to following treatment of diseases: hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, the heterozygous familial hypercholesterolemia, familial hypercholesterolemia isozygotys, the Combination dyslipidemia, atherosclerosis, develop atherosclerotic risk, coronary heart disease, the coronary disease medical history, the coronary heart disease of early showing effect, one or more coronary heart disease risk factors, type ii diabetes, type ii diabetes companion dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipemia, the high fatty acid mass formed by blood stasis, fatty degeneration of liver, non-alcoholic stellato-hepatitis or non-alcohol fatty liver.In other embodiments, treatment is the reduction of CHD risk.In certain embodiments, treatment is atherosclerotic prevention.In certain embodiments, treatment is the prevention of coronary heart disease.

In certain embodiments, the pharmaceutical composition that will comprise the short antisense compounds of targeting ApoB nucleic acid is used for preparing the medicine in order to LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp (a), serum triglycerides, liver tg, Ox-LDL-C, little LDL granule, little VLDL, phospholipid or the oxidized phospholipids that reduces individuality.In certain embodiments, the pharmaceutical composition that will comprise the short antisense compounds of targeting ApoB nucleic acid is used to prepare in order to reduce the medicine of coronary heart disease risk.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid is used to prepare in order to treat the medicine of following disease: hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, the heterozygous familial hypercholesterolemia, familial hypercholesterolemia isozygotys, the Combination dyslipidemia, atherosclerosis, develop atherosclerotic risk, coronary heart disease, the coronary disease medical history, the coronary heart disease of early showing effect, one or more coronary heart disease risk factors, type ii diabetes, type ii diabetes companion dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipemia, the high fatty acid mass formed by blood stasis, fatty degeneration of liver, non-alcoholic stellato-hepatitis or non-alcohol fatty liver.

The ApoB combination treatment

In certain embodiments, the pharmaceutical composition that one or more is comprised the short antisense compounds of targeting ApoB nucleic acid gives altogether with one or more other medicament.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease identical with described one or more pharmaceutical compositions of the present invention.In some this embodiment, described one or more medicaments are that lipid reduces medicament.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease different with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament is designed to treat the ill effect of one or more pharmaceutical compositions of the present invention.In certain embodiments, one or more pharmaceutical compositions of the present invention are given altogether with another medicament, to treat the ill effect of other these another medicaments.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given simultaneously.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given at different time.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is prepared together in single dosage form.In certain embodiments, with other separately preparation of medicament of one or more pharmaceutical compositions of the present invention and one or more.

In certain embodiments, the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting ApoB nucleic acid comprises that lipid reduces medicament.In some this embodiment, can include but not limited to atorvastatin, simvastatin, rosuvastatin with the medicament that pharmaceutical composition of the present invention gives altogether and according to Ezetimibe.In some this embodiment, lipid is reduced medicament before giving the present composition, give.In some this embodiment, lipid is reduced medicament after giving the present composition, give.In some this embodiment, lipid is reduced medicament when giving the present composition, give.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is identical with the dosage that gives separately to be given when lipid reduces medicament.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is lower than the dosage that is given when giving lipid reduction medicament separately.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is higher than the dosage that is given when giving lipid reduction medicament separately.

In certain embodiments, it is the HMG-CoA reductase inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, the HMG-CoA reductase inhibitor is an inhibin.In some this embodiment, inhibin is selected from atorvastatin, simvastatin, pravastatin, fluvastatin and rosuvastatin.

In certain embodiments, it is cholesterol absorption inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, cholesterol absorption inhibitor is according to Ezetimibe.

In certain embodiments, it is HMG-CoA reductase inhibitor and cholesterol absorption inhibitor formulated together altogether that the lipid that gives altogether reduces medicament.In some this embodiment, it is according to Ezetimibe/simvastatin that lipid formulated together altogether reduces medicament.

In certain embodiments, it is microsomal triglyceride transfer protein inhibitor (MTP inhibitor) that the lipid that gives altogether reduces medicament.

In certain embodiments, the medicament that gives altogether is a bile acid chelating agent.In some this embodiment, bile acid chelating agent is selected from colestyramine, colestipol and colesevelam.

In certain embodiments, the medicament that gives altogether is a nicotinic acid.In some this embodiment, nicotinic acid is selected from rapid release nicotinic acid, extended release nicotinic acid and slow release nicotinic acid.

In certain embodiments, the medicament that gives altogether is Carboxymethylcellulose (fibric acid).In some this embodiment, Carboxymethylcellulose is selected from gemfibrozil, fenofibrate, chlorine Bei Te, bezafibrate and ciprofibrate.

More examples of the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting ApoB nucleic acid include but not limited to corticosteroid (including but not limited to prednisone); Immunoglobulin (includes but not limited to vein immunoglobulin (IVIg); Analgesic (for example acetaminophen); Antiinflammatory (including but not limited to NSAID (non-steroidal anti-inflammatory drug) (for example ibuprofen, COX-1 inhibitor and cox 2 inhibitor)); Salicylate; Antibiotic; Antiviral agent; Antifungal; Antidiabetic medicine (for example biguanide, glucosidase inhibitor, insulin, sulfonylureas and thiazolidinediones); The adrenergic modulation agent; Diuretic; Hormone (for example short anabolism steroid, androgen, estrogen, calcitonin, progesterone, somatostatin and thyroxin); Immunomodulator; Muscle relaxant; Hydryllin; Osteoporosis medicament (for example diphosphonate, calcitonin and estrogen); Prostaglandin; Antineoplastic agent; The spiritual healing medicament; Tranquilizer; Poison Oak Tree (oak) or black poison wood (sumac) product; Antibody and vaccine.

In certain embodiments, the pharmaceutical composition that comprises the short antisense compounds of targeting ApoB nucleic acid can be reduced therapy in conjunction with lipid gives.In some this embodiment, it is that curative life style changes that lipid reduces therapy.In some this embodiment, it is the LDL-Apheresis that lipid reduces therapy.

In one embodiment, antisense compounds provided herein can be used to reduce the level of the lipoprotein that contains apolipoprotein B among the people experimenter." lipoprotein that contains apolipoprotein B " used herein refers to any lipoprotein with apolipoprotein B as its protein component, it is believed that to comprise LDL, VLDL, IDL and lipoprotein.LDL, VLDL, IDL and lipoprotein contain the apolipoprotein B of a molecule separately, so serum apolipoprotein B measured value can reflect the total quantity of these lipoproteins.Known in this field, each above-mentioned lipoprotein can both cause atheroma.Therefore, one or more in the reduction serum contain the lipoprotein of apolipoprotein B, the treatment benefit are provided can for people experimenter.Little LDL granule it is believed that than big LDL granule more can cause atheroma, therefore reduces little LDL granule and the treatment benefit is provided can for people experimenter.Other lipid parameter also can determine in the experimenter.T-CHOL: HDL than or LDL: the reduction of HDL ratio is to want the improvement than the aspect of the cholesterol that obtains clinically.Also want to reduce the serum triglycerides of the philtrum that shows that lipid level raises equally, clinically.

Other indications of the cardiovascular disease that can measure in the experimenter comprise the serum LDL granular size; Serum LDL cholesteryl ester concentration; The serum LDL cholesteryl ester is formed; The degree of unsaturation of serum LDL cholesteryl ester; With the serum hdl cholesterol levels.The classification of " serum LDL granular size " used herein ester serum LDL granular size, this classification can comprise minimum, little, medium or bulky grain, represents with g/ μ mol usually.In situation of the present invention, " serum LDL cholesteryl ester concentration " means the quantity that the cholesterol that exists in the LDL granule refers to, measures with mg/dL usually.In situation of the present invention, " serum LDL cholesteryl ester composition " is the percentage measurements of exist in the serum LDL granule saturated, single unsaturated and how unsaturated cholesteryl ester fatty acid." many degrees of unsaturation of serum LDL cholesteryl ester " mean the percentage ratio of the how unsaturated cholesteryl ester fatty acid in the serum LDL granule.

Obtain serum or plasma sample for the method for analyzing with prepare blood serum sample so that the method for analysis is well known to a person skilled in the art.With regard to the measurement of lipoprotein, cholesterol, triglyceride and cholesteryl ester, term " serum " and " blood plasma " can be used in this article mutually.

In another embodiment, antisense compounds provided herein can be used to treat metabolic disease.There is multiple biomarker to can be used to assess metabolic disease.For example, blood glucose levels can be by doctor or even patient with the test kit that obtains usually or blood glucose meter (Ascensia ELITE for example^TMKit, Ascensia (Bayer), Tarrytown NY or Accucheck, Roche Diagnostics) measure.Also can measure (glycated) hemoglobin (HbA of saccharifying_1c).HbA_1cBe a kind of stable less (minor) haemoglobin variant, the post translational modification of being undertaken by glucose forms in vivo, and it contains the NH of remarkable saccharifying₂Terminal β chain.HbA_1cBetween level and preceding 3 months average blood glucose level very strong dependency is arranged.Therefore, HbA_1cOften be regarded as measuring the blood-glucose control of keeping " goldstandard " (Bunn, H.F.et al., 1978, Science.200,21-7).HbA_1cCan measure by ion exchange HPLC or immunoassay; Be used for HbA_1cThe family's blood collecting measured and mailing apparatus (kit) can obtain now everywhere.The serum fructosamine is that another that stablize glucose control measured, and can measure (Cobas Integra, Roche Diagnostics) by colorimetric method.

The short antisense compounds of some targeting ApoB nucleic acid

In certain embodiments, lacking the antisense compounds targeting has

The ApoB nucleic acid of the sequence of searching number NM_000384.1 (being incorporated herein) with SEQ ID NO:1.In some this embodiment, short antisense compounds and the SEQ IDNO:1 of targeting SEQ ID NO:1 have at least 90% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:1 of targeting SEQ ID NO:1 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:1 of targeting SEQ ID NO:1 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from the nucleotide sequence that table 2 and table 3 provide.

The nucleotide sequence that each SEQ ID NO provides in the table 2 and 3 is independent of any modification to sugar moieties, Dan Julian key or nuclear base.Thus, the defined short antisense compounds of certain SEQ ID NO can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 2 and 3 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:1.Table 2 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:1.Table 3 has been enumerated the short antisense compounds that has one or two mispairing with respect to SEQ ID NO:1.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment flank of 10 2 '-deoxyribonucleotides is connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Table 2: the short antisense compounds of targeting SEQ ID NO:1

ISIS No	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						372816	263	278	CCGGAGGTGCTTGAAT	3-10-3 MOE	16
372894	264	277	CGGAGGTGCTTGAA	2-10-2 MOE	17
						372817	428	443	GAAGCCATACACCTCT	3-10-3 MOE	18
372895	429	442	AAGCCATACACCTC	2-10-2 MOE	19
						372818	431	446	GTTGAAGCCATACACC	3-10-3 MOE	20
372896	432	445	TTGAAGCCATACAC	2-10-2 MOE	21
						372819	438	453	CCTCAGGGTTGAAGCC	3-10-3 MOE	22
372897	439	452	CTCAGGGTTGAAGC	2-10-2 MOE	23
						372820	443	458	TTTGCCCTCAGGGTTG	3-10-3 MOE	24
372898	444	457	TTGCCCTCAGGGTT	2-10-2 MOE	25
						372821	468	483	AGTTCTTGGTTTTCTT	3-10-3 MOE	26
372899	469	482	GTTCTTGGTTTTCT	2-10-2 MOE	27
						372822	587	602	CCTCTTGATGTTCAGG	3-10-3 MOE	28
372900	588	601	CTCTTGATGTTCAG	2-10-2 MOE	29
						372823	592	607	ATGCCCCTCTTGATGT	3-10-3 MOE	30
372901	593	606	TGCCCCTCTTGATG	2-10-2 MOE	31
						346583	715	728	TGCCACATTGCCCT	3-8-3 MOE	32
346584	716	729	TTGCCACATTGCCC	3-8-3 MOE	33

346585	717	730	GTTGCCACATTGCC	3-8-3 MOE	34
						346586	718	731	TGTTGCCACATTGC	3-8-3 MOE	35
346587	719	732	CTGTTGCCACATTG	3-8-3 MOE	36
						346588	720	733	TCTGTTGCCACATT	3-8-3 MOE	37
346589	721	734	TTCTGTTGCCACAT	3-8-3 MOE	38
						346590	722	735	TTTCTGTTGCCACA	3-8-3 MOE	39
346591	723	736	ATTTCTGTTGCCAC	3-8-3 MOE	40
						372824	929	944	GTAGGAGAAAGGCAGG	3-10-3 MOE	41
372902	930	943	TAGGAGAAAGGCAG	2-10-2 MOE	42
						372825	1256	1271	GGCTTGTAAAGTGATG	3-10-3 MOE	43
372903	1257	1270	GCTTGTAAAGTGAT	2-10-2 MOE	44
						372826	1304	1319	CCACTGGAGGATGTGA	3-10-3 MOE	45
372904	1305	1318	CACTGGAGGATGTG	2-10-2 MOE	46
						372829	2135	2150	TTTCAGCATGCTTTCT	3-10-3 MOE	47
372907	2136	2149	TTCAGCATGCTTTC	2-10-2 MOE	48
						372832	2774	2789	CATATTTGTCACAAAC	3-10-3 MOE	49
372910	2775	2788	ATATTTGTCACAAA	2-10-2 MOE	50
						372833	2779	2794	ATGCCCATATTTGTCA	3-10-3 MOE	51
372911	2780	2793	TGCCCATATTTGTC	2-10-2 MOE	52
						372835	2961	2976	TTTTGGTGGTAGAGAC	3-10-3 MOE	53
372913	2962	2975	TTTGGTGGTAGAGA	2-10-2 MOE	54
						346592	3248	3261	TCTGCTTCGCACCT	3-8-3 MOE	55
346593	3249	3262	GTCTGCTTCGCACC	3-8-3 MOE	56
						346594	3250	3263	AGTCTGCTTCGCAC	3-8-3 MOE	57
346595	3251	3264	CAGTCTGCTTCGCA	3-8-3 MOE	58
						346596	3252	3265	TCAGTCTGCTTCGC	3-8-3 MOE	59
346597	3253	3266	CTCAGTCTGCTTCG	3-8-3 MOE	60
						346598	3254	3267	CCTCAGTCTGCTTC	3-8-3 MOE	61
346599	3255	3268	GCCTCAGTCTGCTT	3-8-3 MOE	62
						346600	3256	3269	AGCCTCAGTCTGCT	3-8-3 MOE	63
372836	3350	3365	AACTCTGAGGATTGTT	3-10-3 MOE	64
						372914	3351	3364	ACTCTGAGGATTGT	2-10-2 MOE	65
372837	3355	3370	TCATTAACTCTGAGGA	3-10-3 MOE	66
						372915	3356	3369	CATTAACTCTGAGG	2-10-2 MOE	67
372838	3360	3375	ATTCATCATTAACTCT	3-10-3 MOE	68
						372916	3361	3374	TTCATCATTAACTC	2-10-2 MOE	69
372839	3409	3424	TTGTTCTGAATGTCCA	3-10-3 MOE	70
						387461	3409	3424	TTGTTCTGAATGTCCA	Cytosine in the 3-10-3 methylene oxygen base BNA interval is not modified	70
380147	3409	3424	TTGTTCTGAATGTCCA	3-10-3 methylene oxygen base BNA	70
						372917	3410	3423	TGTTCTGAATGTCC	2-10-2 MOE	73
372840	3573	3588	CAGATGAGTCCATTTG	3-10-3 MOE	74
						372918	3574	3587	AGATGAGTCCATTT	2-10-2 MOE	75
372841	3701	3716	ATCCACAGGGAAATTG	3-10-3 MOE	76
						372919	3702	3715	TCCACAGGGAAATT	2-10-2 MOE	77
372843	4219	4234	CAGTTGTACAAGTTGC	3-10-3 MOE	78
						372921	4220	4233	AGTTGTACAAGTTG	2-10-2 MOE	79
372844	4301	4316	CACAGAGTCAGCCTTC	3-10-3 MOE	80

372922	4302	4315	ACAGAGTCAGCCTT	2-10-2 MOE	81
						372845	4308	4323	GGTCAACCACAGAGTC	3-10-3 MOE	82
372923	4309	4322	GTCAACCACAGAGT	2-10-2 MOE	83
						346601	5588	5601	CAGCCACATGCAGC	3-8-3 MOE	84
346602	5589	5602	CCAGCCACATGCAG	3-8-3 MOE	85
						346603	5590	5603	ACCAGCCACATGCA	3-8-3 MOE	86
346604	5591	5604	TACCAGCCACATGC	3-8-3 MOE	87
						346605	5592	5605	TTACCAGCCACATG	3-8-3 MOE	88
346606	5593	5606	GTTACCAGCCACAT	3-8-3 MOE	89
						346607	5594	5607	GGTTACCAGCCACA	3-8-3 MOE	90
346608	5595	5608	AGGTTACCAGCCAC	3-8-3 MOE	91
						346609	5596	5609	TAGGTTACCAGCCA	3-8-3 MOE	92
372851	5924	5939	AGGTTCTGCTTTCAAC	3-10-3 MOE	93
						372929	5925	5938	GGTTCTGCTTTCAA	2-10-2 MOE	94
372854	6664	6679	TACTGATCAAATTGTA	3-10-3 MOE	95
						372932	6665	6678	ACTGATCAAATTGT	2-10-2 MOE	96
372855	6908	6923	TTTTTCTTGTATCTGG	3-10-3 MOE	97
						372933	6909	6922	TTTTCTTGTATCTG	2-10-2 MOE	98
372856	7190	7205	ATCCATTAAAACCTGG	3-10-3 MOE	99
						372934	7191	7204	TCCATTAAAACCTG	2-10-2 MOE	100
372858	7817	7832	ATATTGCTCTGCAAAG	3-10-3 MOE	101
						372936	7818	7831	TATTGCTCTGCAAA	2-10-2 MOE	102
346610	7818	7831	TATTGCTCTGCAAA	3-8-3 MOE	102
						346611	7819	7832	ATATTGCTCTGCAA	3-8-3 MOE	104
346612	7820	7833	AATATTGCTCTGCA	3-8-3 MOE	105
						346613	7821	7834	GAATATTGCTCTGC	3-8-3 MOE	106
346614	7822	7835	AGAATATTGCTCTG	3-8-3 MOE	107
						346615	7823	7836	TAGAATATTGCTCT	3-8-3 MOE	108
346616	7824	7837	ATAGAATATTGCTC	3-8-3 MOE	109
						346617	7825	7838	GATAGAATATTGCT	3-8-3 MOE	110
346618	7826	7839	GGATAGAATATTGC	3-8-3 MOE	111
						372859	7995	8010	ATGGAATCCTCAAATC	3-10-3 MOE	112
372937	7996	8009	TGGAATCCTCAAAT	2-10-2 MOE	113
						372861	8336	8351	GAATTCTGGTATGTGA	3-10-3 MOE	114
372939	8337	8350	AATTCTGGTATGTG	2-10-2 MOE	115
						372862	8341	8356	AGCTGGAATTCTGGTA	3-10-3 MOE	116
372940	8342	8355	GCTGGAATTCTGGT	2-10-2 MOE	117
						372863	8539	8554	TGAAAATCAAAATTGA	3-10-3 MOE	118
372941	8540	8553	GAAAATCAAAATTG	2-10-2 MOE	119
						372871	9344	9359	AAACAGTGCATAGTTA	3-10-3 MOE	120
372949	9345	9358	AACAGTGCATAGTT	2-10-2 MOE	121
						372872	9515	9530	TTCAGGAATTGTTAAA	3-10-3 MOE	122
372950	9516	9529	TCAGGAATTGTTAA	2-10-2 MOE	123
						372875	9794	9809	TTTTGTTTCATTATAG	3-10-3 MOE	124
372953	9795	9808	TTTGTTTCATTATA	2-10-2 MOE	125
						372877	10157	10172	GATGACACTTGATTTA	3-10-3 MOE	126
372955	10158	10171	ATGACACTTGATTT	2-10-2 MOE	127
						372878	10161	10176	GTGTGATGACACTTGA	3-10-3 MOE	128
372956	10162	10175	TGTGATGACACTTG	2-10-2 MOE	129
						372879	10167	10182	TATTCAGTGTGATGAC	3-10-3 MOE	130
372957	10168	10181	ATTCAGTGTGATGA	2-10-2 MOE	131

372880	10172	10187	ATTGGTATTCAGTGTG	3-10-3 MOE	132
						372958	10173	10186	TTGGTATTCAGTGT	2-10-2 MOE	133
346619	10838	10851	CCTCTAGCTGTAAG	3-8-3 MOE	134
						346620	10839	10852	CCCTCTAGCTGTAA	3-8-3 MOE	135
346621	10840	10853	GCCCTCTAGCTGTA	3-8-3 MOE	136
						346622	10841	10854	GGCCCTCTAGCTGT	3-8-3 MOE	137
346623	10842	10855	AGGCCCTCTAGCTG	3-8-3 MOE	138
						346624	10843	10856	GAGGCCCTCTAGCT	3-8-3 MOE	139
346625	10844	10857	AGAGGCCCTCTAGC	3-8-3 MOE	140
						346626	10845	10858	AAGAGGCCCTCTAG	3-8-3 MOE	141
346627	10846	10859	AAAGAGGCCCTCTA	3-8-3 MOE	142
						372890	13689	13704	GAATGGACAGGTCAAT	3-10-3 MOE	143
372968	13690	13703	AATGGACAGGTCAA	2-10-2 MOE	144
						372891	13694	13709	GTTTTGAATGGACAGG	3-10-3 MOE	145
372969	13695	13708	TTTTGAATGGACAG	2-10-2 MOE	146
						372892	13699	13714	TGGTAGTTTTGAATGG	3-10-3 MOE	147
372970	13700	13713	GGTAGTTTTGAATG	2-10-2 MOE	148
						346628	13907	13920	TCACTGTATGGTTT	3-8-3 MOE	149
346629	13908	13921	CTCACTGTATGGTT	3-8-3 MOE	150
						346630	13909	13922	GCTCACTGTATGGT	3-8-3 MOE	151
346631	13910	13923	GGCTCACTGTATGG	3-8-3 MOE	152
						346632	13911	13924	TGGCTCACTGTATG	3-8-3 MOE	153
346633	13912	13925	CTGGCTCACTGTAT	3-8-3 MOE	154
						346634	13913	13926	GCTGGCTCACTGTA	3-8-3 MOE	155
346635	13914	13927	GGCTGGCTCACTGT	3-8-3 MOE	156
						346636	13915	13928	AGGCTGGCTCACTG	3-8-3 MOE	157
346637	13963	13976	CAGGTCCAGTTCAT	3-8-3 MOE	158
						346638	13964	13977	GCAGGTCCAGTTCA	3-8-3 MOE	159
346639	13965	13978	TGCAGGTCCAGTTC	3-8-3 MOE	160
						346640	13966	13979	GTGCAGGTCCAGTT	3-8-3 MOE	161
346641	13967	13980	GGTGCAGGTCCAGT	3-8-3 MOE	162
						346642	13968	13981	TGGTGCAGGTCCAG	3-8-3 MOE	163
346643	13969	13982	TTGGTGCAGGTCCA	3-8-3 MOE	164
						346644	13970	13983	TTTGGTGCAGGTCC	3-8-3 MOE	165
346645	13971	13984	CTTTGGTGCAGGTC	3-8-3 MOE	166
						346646	14051	14064	TAACTCAGATCCTG	3-8-3 MOE	167
346647	14052	14065	ATAACTCAGATCCT	3-8-3 MOE	168
						346648	14053	14066	AATAACTCAGATCC	3-8-3 MOE	169
346649	14054	14067	AAATAACTCAGATC	3-8-3 MOE	170
						346650	14055	14068	AAAATAACTCAGAT	3-8-3 MOE	171
346651	14056	14069	CAAAATAACTCAGA	3-8-3 MOE	172
						346652	14057	14070	GCAAAATAACTCAG	3-8-3 MOE	173
346653	14058	14071	AGCAAAATAACTCA	3-8-3 MOE	174
						346654	14059	14072	TAGCAAAATAACTC	3-8-3 MOE	175

Table 3: targeting SEQ ID NO:1 and have the short antisense compounds of 1 or 2 mispairing

Isis NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						372894	771	784	CGGAGGTGCTTGAA	2-10-2 MOE	17
372905	1111	1124	CAGGGCCTGGAGAG	2-10-2 MOE	176
						346628	1493	1506	TCACTGTATGGTTT	3-8-3 MOE	149
372828	2006	2021	TCTGAAGTCCATGATC	3-10-3 MOE	177
						372906	2007	2020	CTGAAGTCCATGAT	2-10-2 MOE	178
372830	2382	2397	TGGGCATGATTCCATT	3-10-3 MOE	179
						372908	2383	2396	GGGCATGATTCCAT	2-10-2 MOE	180
346616	3162	3175	ATAGAATATTGCTC	3-8-3 MOE	109
						346617	3163	3176	GATAGAATATTGCT	3-8-3 MOE	110
372929	3513	3526	GGTTCTGCTTTCAA	2-10-2 MOE	94
						372946	3800	3813	TGGAGCCCACGTGC	2-10-2 MOE	181
372904	4040	4053	CACTGGAGGATGTG	2-10-2 MOE	46
						372842	4084	4099	TTGAAGTTGAGGGCTG	3-10-3 MOE	182
372920	4085	4098	TGAAGTTGAGGGCT	2-10-2 MOE	183
						346586	4778	4791	TGTTGCCACATTGC	3-8-3 MOE	35
372847	5030	5045	ACCAGTATTAATTTTG	3-10-3 MOE	184
						372925	5031	5044	CCAGTATTAATTTT	2-10-2 MOE	185
372848	5192	5207	GTGTTCTTTGAAGCGG	3-10-3 MOE	186
						372926	5193	5206	TGTTCTTTGAAGCG	2-10-2 MOE	187
372953	5625	5638	TTTGTTTCATTATA	2-10-2 MOE	125
						372935	7585	7598	AGTTACTTTGGTGT	2-10-2 MOE	188
372860	8255	8270	TGGTACATGGAAGTCT	3-10-3 MOE	189
						372938	8256	8269	GGTACATGGAAGTC	2-10-2 MOE	190
391260	8256	8269	GGTACATGGAAGTC	2-10-2 MOE	190
						392068	8256	8269	GGTACATGGAAGTC	2-10-2 MOE	190
387462	8256	8269	GGTACATGGAAGTC	2-10-2 methylene oxygen base BNA	190
						391872	8256	8269	GGTACATGGAAGTC	Cytosine in 1-1-10-2 2 '-(butyl acetamido)-palmitamide methylene oxygen base BNA/ methylene oxygen base BNA interval is not modified	190
380148	8256	8269	GGTACATGGAAGTC	2-10-2 methylene oxygen base BNA	190
						391871	8256	8269	GGTACATGGAAGTC	Cytosine in 1-1-10-2 2 '-(butyl acetamido)-palmitamide/MOE/MOE interval is not modified	190
391755	8256	8269	GGTACATGGAAGTC	2-10-2ENA mC is only in the pterion	190
						398296	8256	8269	GGTACATGGAAGTC	2-10-2 (6 ' S)-6 '-methyl-methylene oxygen base	190

				The BNA cytosine is not modified
						372942	8455	8468	TCCATGCCATATGT	2-10-2 MOE	200
372865	8888	8903	CCCTGAAGAAGTCCAT	3-10-3 MOE	201
						372943	8889	8902	CCTGAAGAAGTCCA	2-10-2 MOE	202
372866	8908	8923	GCCCAGTTCCATGACC	3-10-3 MOE	203
						372944	8909	8922	CCCAGTTCCATGAC	2-10-2 MOE	204
372867	9058	9073	TTGAGGAAGCCAGATT	3-10-3 MOE	205
						372945	9059	9072	TGAGGAAGCCAGAT	2-10-2 MOE	206
372870	9261	9276	TGGATGCAGTAATCTC	3-10-3 MOE	207
						372948	9262	9275	GGATGCAGTAATCT	2-10-2 MOE	208
372881	10185	10200	TATAAAGTCCAGCATT	3-10-3 MOE	209
						372959	10186	10199	ATAAAGTCCAGCAT	2-10-2 MOE	210
372882	10445	10460	AAGTTCCTGCTTGAAG	3-10-3 MOE	211
						372960	10446	10459	AGTTCCTGCTTGAA	2-10-2 MOE	212
372964	11451	11464	AATGGTGAAGTACT	2-10-2 MOE	213
						346612	13459	13472	AATATTGCTCTGCA	3-8-3 MOE	105
346613	13460	13473	GAATATTGCTCTGC	3-8-3 MOE	106

In certain embodiments, the target zone is the nucleotide 263-278 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 263-278 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO:16 or 17.In some this embodiment, the short antisense compounds of the nucleotide 263-278 of targeting SEQ ID NO:1 is selected from Isis NO.372816 or 372894.

In certain embodiments, the target zone is the nucleotide 428-483 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 428-483 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 18,19,20,21,22,23,24,25,26 or 27.In some this embodiment, the short antisense compounds of the nucleotide 428-483 of targeting SEQ ID NO:1 is selected from Isis NO.372817,372895,372818,372896,372819,372897,372820,372898,372821 or 372899.

In certain embodiments, the target zone is the nucleotide 428-458 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 428-458 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 18,19,20,21,22,23,24 or 25.In some this embodiment, the short antisense compounds of the nucleotide 428-458 of targeting SEQ ID NO:1 is selected from Isis NO.372817,372895,372818,372896,372819,372897,372820 or 372898.

In certain embodiments, the target zone is the nucleotide 468-483 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 468-483 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 26 or 27.In some this embodiment, the short antisense compounds of the nucleotide 468-483 of targeting SEQ ID NO:1 is selected from Isis NO.372821 or 372899.

In certain embodiments, the target zone is the nucleotide 587-607 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 587-607 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 28,29,30 or 31.In some this embodiment, the short antisense compounds of the nucleotide 587-607 of targeting SEQ ID NO:1 is selected from ISIS NO.372822,372900,372823 or 372901.

In certain embodiments, the target zone is the nucleotide 715-736 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 715-736 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 32,33,34,35,36,37,38,39 or 40.In some this embodiment, the short antisense compounds of the nucleotide 715-736 of targeting SEQ ID NO:1 is selected from Isis NO.346583,346584,346585,346586,346587,346588,346589,346590 or 346591.

In certain embodiments, the target zone is the nucleotide 929-944 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 929-944 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 41 or 42.In some this embodiment, the short antisense compounds of the nucleotide 929-944 of targeting SEQ ID NO:1 is selected from Isis NO.372824 or 372902.

In certain embodiments, the target zone is the nucleotide 1256-1319 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 1256-1319 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 43,44,45 or 46.In some this embodiment, the short antisense compounds of the nucleotide 1256-1319 of targeting SEQ ID NO:1 is selected from Isis NO.372825,372903,372826 or 372904.

In certain embodiments, the target zone is the nucleotide 1256-1271 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 1256-1271 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 43 or 44.In some this embodiment, the short antisense compounds of the nucleotide 1256-1271 of targeting SEQ ID NO:1 is selected from Isis NO.372825 or 372903.

In certain embodiments, the target zone is the nucleotide 1304-1319 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 1304-1319 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 45 or 46.In some this embodiment, the short antisense compounds of the nucleotide 1304-1319 of targeting SEQ ID NO:1 is selected from Isis NO.372826 or 372904.

In certain embodiments, the target zone is the nucleotide 2135-2150 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 2135-2150 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 47 or 48.In some this embodiment, the short antisense compounds of the nucleotide 2135-2150 of targeting SEQ ID NO:1 is selected from ISIS NO.372829 or 372907.

In certain embodiments, the target zone is the nucleotide 2774-2794 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 2774-2794 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 49,50,51 or 52.In some this embodiment, the short antisense compounds of the nucleotide 2774-2794 of targeting SEQ ID NO:1 is selected from ISIS NO.372832,372910,372833 or 372911.

In certain embodiments, the target zone is the nucleotide 2961-2976 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 2961-2976 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 53 or 54.In some this embodiment, the short antisense compounds of the nucleotide 2961-2976 of targeting SEQ ID NO:1 is selected from ISIS NO.372835 or 372913.

In certain embodiments, the target zone is the nucleotide 3248-3269 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 3248-3269 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 55,56,57,58,59,60,61,62 or 63.In some this embodiment, the short antisense compounds of the nucleotide 3248-3269 of targeting SEQID NO:1 is selected from ISIS NO.346592,346593,346594,346595,346596,346597,346598,346599 or 346600.

In certain embodiments, the target zone is the nucleotide 3350-3375 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 3350-3375 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 64,65,66,67,68 or 69.In some this embodiment, the short antisense compounds of the nucleotide 3350-3375 of targeting SEQ ID NO:1 is selected from ISIS NO.372836,372914,372837,372915,372838 or 372916.

In certain embodiments, the target zone is the nucleotide 3409-3424 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 3409-3424 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 70 or 73.In some this embodiment, the short antisense compounds of the nucleotide 3409-3424 of targeting SEQ ID NO:1 is selected from ISIS NO.372839,387461,380147 or 372917.

In certain embodiments, the target zone is the nucleotide 3573-3588 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 3573-3588 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 74 or 75.In some this embodiment, the short antisense compounds of the nucleotide 3573-3588 of targeting SEQ ID NO:1 is selected from ISIS NO.372840 or 372918.

In certain embodiments, the target zone is the nucleotide 3701-3716 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 3701-3716 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 76 or 77.In some this embodiment, the short antisense compounds of the nucleotide 3701-3716 of targeting SEQ ID NO:1 is selected from ISIS NO.372841 or 372919.

In certain embodiments, the target zone is the nucleotide 4219-4234 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 4219-4234 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 78 or 79.In some this embodiment, the short antisense compounds of the nucleotide 4219-4234 of targeting SEQ ID NO:1 is selected from ISIS NO.372843 or 372921.

In certain embodiments, the target zone is the nucleotide 4301-4323 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 4301-4323 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 80,81,82 or 83.In some this embodiment, the short antisense compounds of the nucleotide 4301-4323 of targeting SEQ ID NO:1 is selected from ISIS NO.372844,372922,372845 or 372923.

In certain embodiments, the target zone is the nucleotide 5588-5609 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 5588-5609 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 84,85,86,87,88,89,90,91 or 92.In some this embodiment, the short antisense compounds of the nucleotide 5588-5609 of targeting SEQID NO:1 is selected from ISIS NO.346601,346602,346603,346604,346605,346606,346607,346608 or 346609.

In certain embodiments, the target zone is the nucleotide 5924-5939 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 5924-5939 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 93 or 94.In some this embodiment, the short antisense compounds of the nucleotide 5924-5939 of targeting SEQ ID NO:1 is selected from ISIS NO.372851 or 372929.

In certain embodiments, the target zone is the nucleotide 6664-6679 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 6664-6679 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 95 or 96.In some this embodiment, the short antisense compounds of the nucleotide 6664-6679 of targeting SEQ ID NO:1 is selected from ISIS NO.372854 or 372932.

In certain embodiments, the target zone is the nucleotide 6908-6923 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 6908-6923 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 97 or 98.In some this embodiment, the short antisense compounds of the nucleotide 6908-6923 of targeting SEQ ID NO:1 is selected from ISIS NO.372855 or 372933.

In certain embodiments, the target zone is the nucleotide 7190-7205 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 7190-7205 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 99 or 100.In some this embodiment, the short antisense compounds of the nucleotide 7190-7205 of targeting SEQ ID NO:1 is selected from ISIS NO.372856 or 372934.

In certain embodiments, the target zone is the nucleotide 7817-7839 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 7817-7839 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 101,102,104,105,106,107,108,109,110 or 111.In some this embodiment, the short antisense compounds of the nucleotide 7817-7839 of targeting SEQ ID NO:1 is selected from ISIS NO.372858,372936,346610,346611,346612,346613,346614,346615,346616,346617 or 346618.

In certain embodiments, the target zone is the nucleotide 7995-8010 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 7995-8010 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 112 or 113.In some this embodiment, the short antisense compounds of the nucleotide 7995-8010 of targeting SEQ ID NO:1 is selected from ISIS NO.372859 or 372937.

In certain embodiments, the target zone is the nucleotide 8336-8356 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 8336-8356 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 114,115,116 or 117.In some this embodiment, the short antisense compounds of the nucleotide 8336-8356 of targeting SEQ ID NO:1 is selected from ISIS NO.372861,372939,372862 or 372940.

In certain embodiments, the target zone is the nucleotide 8539-8554 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 8539-8554 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 118 or 119.In some this embodiment, the short antisense compounds of the nucleotide 8539-8554 of targeting SEQ ID NO:1 is selected from ISIS NO.372863 or 372941.

In certain embodiments, the target zone is the nucleotide 9344-9359 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 9344-9359 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 120 or 121.In some this embodiment, the short antisense compounds of the nucleotide 9344-9359 of targeting SEQ ID NO:1 is selected from ISIS NO.372871 or 372949.

In certain embodiments, the target zone is the nucleotide 9515-9530 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 9515-9530 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 122 or 123.In some this embodiment, the short antisense compounds of the nucleotide 9515-9530 of targeting SEQ ID NO:1 is selected from ISIS NO.372872 or 372950.

In certain embodiments, the target zone is the nucleotide 9794-9809 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 9794-9809 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 124 or 125.In some this embodiment, the short antisense compounds of the nucleotide 9794-9809 of targeting SEQ ID NO:1 is selected from ISIS NO.372875 or 372953.

In certain embodiments, the target zone is the nucleotide 10157-10187 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 10157-10187 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 126,127,128,129,130,131,132 or 133.In some this embodiment, the short antisense compounds of the nucleotide 10157-10187 of targeting SEQ ID NO:1 is selected from ISISNO.372877,372955,372878,372956,372879,372957,372880 or 372958.

In certain embodiments, the target zone is the nucleotide 10838-10859 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 10838-10859 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 134,135,136,137,138,139,140,141 or 142.In some this embodiment, the short antisense compounds of the nucleotide 10838-10859 of targeting SEQ ID NO:1 is selected from ISIS NO.346619,346620,346621,346622,346623,346624,346625,346626 or 346627.

In certain embodiments, the target zone is the nucleotide 13689-13714 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 13689-13714 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 143,144,145,146,147 or 148.In some this embodiment, the short antisense compounds of the nucleotide 13689-13714 of targeting SEQ IDNO:1 is selected from ISIS NO.372890,372968,372891,372969,372892 or 372970.

In certain embodiments, the target zone is the nucleotide 13907-13928 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 13907-13928 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 149,150,151,152,153,154,155,156 or 157.In some this embodiment, the short antisense compounds of the nucleotide 13907-13928 of targeting SEQ ID NO:1 is selected from ISIS NO.346628,346629,346630,346631,346632,346633,346634,346635 or 346636.

In certain embodiments, the target zone is the nucleotide 13963-13984 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 13963-13984 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 158,159,160,161,162,163,164,165 or 166.In some this embodiment, the short antisense compounds of the nucleotide 13963-13984 of targeting SEQ ID NO:1 is selected from ISIS NO.346637,346638,346639,346640,346641,346642,346643,346644 or 346645.

In certain embodiments, the target zone is the nucleotide 14051-14072 of SEQ ID NO:1.In some this embodiment, the short antisense compounds of the nucleotide 14051-14072 of targeting SEQ ID NO:1 comprises the nucleotide sequence that is selected from SEQ ID NO 167,168,169,170,171,172,173,174 or 175.In some this embodiment, the short antisense compounds of the nucleotide 14051-14072 of targeting SEQ ID NO:1 is selected from ISIS NO.346646,346647,346648,346649,346650,346651,346652,346653 or 346654.

In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting ApoB nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) or O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting ApoB nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating ApoB.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting ApoB nucleic acid, and wherein the short antisense compounds of this targeting ApoB nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting ApoB nucleic acid is regulated the expression of ApoB.

In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 8 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 9 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 10 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 11 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 12 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 13 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 14 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 15 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for adjusting ApoB comprises that using length is the short antisense compounds of 16 monomeric targeting ApoB nucleic acid.

In certain embodiments, the method for the expression of adjusting ApoB comprises that use comprises the short antisense compounds of 9-15 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for the expression of adjusting ApoB comprises that use comprises the short antisense compounds of 10-15 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for the expression of adjusting ApoB comprises that use comprises the short antisense compounds of 12-14 monomeric targeting ApoB nucleic acid.In certain embodiments, the method for the expression of adjusting ApoB comprises the short antisense compounds that uses the targeting ApoB nucleic acid that comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid can have short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting ApoB nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3 and 1-9-2.

2.SGLT-2

Sodium dependent glucose transport protein 2 (SGLT-2) is expressed in kidney proximal tubule epithelial cell, absorbs glucose again and prevents the effect of loss of glucose in urine thereby play.For the people's gene group, SGLT-2 is a member in the 11 member families of sodium substrate cotransporter.Many these family members enjoy sequence homology, and for example SGLT-1 and SGLT-2 enjoy about 59% sequence homogeneity, enjoy about 70% sequence homogeneity with SGLT-3.SGLT-1 is the glucose transporter that is present among heart and the CNS.SGLT-3 is the sodium channel of experiencing glucose in the small intestinal.The separation station-keeping mode of these SGLTs is a difference part between these homologies family member.(Handlon，A.L.，Expert Opin.Ther.Patents(2005)15(11)：1532-1540；Kanai et al.，J.Clin.Invest.，1994，93，397-404；Wellset al.，Am.J.Physiol.Endocrinol.Metab.，1992，263，F459-465)。

To being expelled to studies have shown that of people SGLT2 in Xenopus laevis (Xenopus) oocyte, this albumen mass-energy mediation D-glucose and Alpha-Methyl-D-pyranglucoside (α-MeGlc; A kind of glucalogue) sodium dependency transhipment, wherein the Km value to α-MeGlc is 1.6mM, sodium and glucose coupling ratio are 1: 1 (Kanai et al., J.Clin.Invest., 1994,93,397-404; You et al., J.Biol.Chem., 1995,270,29365-29371).This transport activity is suppressed by phlorhizin, and phlorhizin is a kind of plant glucosides, can be in conjunction with the glucose site of SGLTs but do not transported, thereby suppress the SGLT effect (You et al., J.Biol.Chem., 1995,270,29365-29371).

Diabetes are to be the disease of feature with the hyperglycemia due to the insulin action deficiency.Chronic hyperglycemia is the major risk factors that comprises the diabetes related complication of heart disease, retinopathy, nephropathy and neuropathy.Because kidney plays important effect in the adjusting of plasma glucose levels, the kidney glucose transporter just becoming noticeable drug targets (Wright, Am.J.Physiol.Renal Physiol., 2001,280, F10-18).Diabetic nephropathy is the common cause of the end-stage renal disease that develops in many diabeticss.The glucose toxicity that causes by long-term hyperglycemia can in diabetics, cause organize the dependency insulin resistance (Nawano et al., Am.J.Physiol.Endocrinol.Metab., 2000,278, E535-543).

Definition

" sodium dependent glucose transport protein 2 " is that its expression is treated by lacking gene outcome or the protein that antisense compounds is regulated.Sodium dependent glucose transport protein 2 is commonly referred to SGLT2, but also can be described as SLC5A2; Sodium-glucose transporter 2; Sodium glucose co-transporter 2 is white, the kidney low-affinity; Cotransporter, kidney; Solute carrier family 5 (sodium/glucose transporter), the member 2; SL52.

" SGLT2 nucleic acid " means the nucleic acid of any coding SGLT2.For example, in certain embodiments, SGLT2 nucleic acid include but not limited to the to encode DNA sequence of SGLT2, the RNA sequence of transcribing from the DNA of coding SGLT2 and the mRNA sequence of coding SGLT2." SGLT2mRNA " means the proteic mRNA of coding SGLT2.

The treatment indication

In certain embodiments, lack the expression that antisense compounds is used for regulating SGLT-2 and related protein.In certain embodiments, this adjusting is can include but not limited to that the short antisense compounds of the target nucleic acids molecular hybridization of following material realizes with one or more codings by providing: SGLT2, SL52, SLC5A2, sodium glucose co-transporter 2 is white, kidney low-affinity sodium glucose co-transporter 2 is white, kidney sodium glucose co-transporter 2 white 2 and solute carrier family 5 sodium/glucose transporter member 2.Also provide the method for treatment metabolic disease as herein described and/or cardiovascular disease and disease.In specific embodiment, the short antisense compounds that can suppress the expression of SGLT2 be used for reducing in the method for blood glucose levels of animal and postpone or the method for the outbreak of prevention type ii diabetes in.This method comprises that one or more The compounds of this invention with treatment or prevention effective dose to need to give the animal for the treatment of.These one or more chemical compounds can be the short antisense compounds of the nucleic acid of targeting coding SGLT2.This paper provides the method for the inhibition of the SGLT2 expression that strengthens in kidney cell or the renal tissue, and described method comprises that the short antisense compounds of the nucleic acid that makes this cell or tissue and one or more The compounds of this invention such as targeting coding SGLT2 contacts.

Though described some chemical compound, compositions and method specially according to some embodiment, following example only is a chemical compound in order to demonstrate the invention, and is not intended to and limits these chemical compounds.

In certain embodiments, lacking antisense compounds is the chimeric oligomeric compounds with mix sulphur substituted phosphate and phosphodiester backbone.The short antisense compounds of some mixed matrix has the central authorities interval that comprises at least 5 adjacency 2 '-deoxynucleoside, and this interval flank is connected to two pterions, and each pterion comprises at least 12 '-O-methoxy ethyl nucleoside.In certain embodiments, connecting key between the nucleoside of mixed matrix chemical compound, is that thiophosphate connects key in the interval, is that di-phosphate ester connects key in two pterions.In certain embodiments, the mixed matrix chemical compound has thiophosphate and connects key in the pterion, exception be oligonucleotide 5 ' and 3 ' terminal one or simultaneously two ends have 1 di-phosphate ester to connect key.In certain embodiments, the short antisense compounds of targeting SGLT2 has the 3-10-3 of being selected from, 2-10-3,2-10-2,1-10-1, the die body of 1-10-2,2-8-2,1-9-2,1-8-1,3-6-3 or 1-6-1 (pterion-deoxidation interval-pterion).In certain embodiments, the short antisense compounds of targeting SGLT2 has the die body (pterion-deoxidation interval-pterion) that is selected from 1-10-1,1-10-2,2-8-2,1-9-2,1-8-1,3-6-3 or 1-6-1.

In certain embodiments, targeting SGLT2 nucleic acid and short antisense compounds with mixed matrix effectively are delivered in the kidney.In certain embodiments, the short antisense compounds that gives targeting SGLT2 nucleic acid and have a mixed matrix can cause the target gene in the kidney to express adjusted.In some this embodiment, liver or Toxicity of Kidney are few or do not have.In certain embodiments, targeting SGLT2 nucleic acid and short antisense compounds with mixed matrix with compare except not having the identical short antisense compounds in external other aspects of mixture skeleton, having more usefulness aspect the reduction SGLT-2mRNA, and onset is faster.In some this embodiment, the usefulness of this raising and/or the toxicity of reduction are in mice and/or rat.In some this embodiment, the usefulness of this raising and/or the toxicity of reduction are in the people.

For example and only for purposes of illustration, only comprising thiophosphate and connect the ISIS145733 of key and comprise di-phosphate ester connect key and comprise the ISIS 257016 that thiophosphate connects key in the interval in the pterion, is identical in other respects.The both comprises sequence GAAGTAGCCACCAACTGTGC (SEQ ID NO.1572).These two oligonucleotide also comprise the interval of being made up of 10 2 '-deoxyribonucleotides, and every side joint at interval has 52 '-methoxy ethyl (2 '-MOE) nucleotide.All cytidine residues all are the 5-methylcytidines.Mixed matrix Compound I SIS 257016 compares with non-mixing parent compound ISIS 145733, is having more usefulness (referring to embodiment 9) aspect the reduction SGLT-2mRNA.

Pharmacokinetic to a certain mixed matrix Compound I SIS 257016 shows that in certain embodiments, this chemical compound plays the effect of prodrug, can be metabolised to the pharmacophore of 12 nuclear bases.To studies show that of this short antisense compounds of 12 nuclear bases corresponding to ISIS 257016 of ISIS 370717, this chemical compound has the pharmacological action (pharmacological profile) similar to ISIS 257016, but onset is faster.ISIS 370717 is 12 nuclear base antisense oligonucleotides of targeting SGLT-2, comprises sequence TAGCCACCAACT (SEQ ID NO.1554), also comprises the interval of being made up of 10 2 '-deoxyribonucleotides, and both sides respectively are connected to the pterion of 1 nucleotide at interval.(2 '-MOE) nucleotide is formed by 2 '-methoxy ethyl in the pterion.All cytidine residues all are the 5-methylcytidines.In whole oligonucleotide, connecting key between nucleoside is thiophosphate (P=S).The similarity of the pharmacological activity of ISIS 257016 and ISIS 370717 has been supported this pharmacokinetic result, and promptly ISIS 257016 is the prodrugs (referring to embodiment 10) with 12 nucleotide pharmacophore.In addition, to the great loss of activity that studies show that of stabilisation (end the adds medicated cap) version of ISIS 257016.

In certain embodiments, comprise the monomeric short antisense compounds of 2 ' MOE and be delivered to effectively in the kidney in the pterion, the treatment of carrying out with this chemical compound causes in the kidney target gene expression effectively to be regulated, and does not have liver or Toxicity of Kidney.This paper is proof also further, and in certain embodiments, the parent oligonucleotide of the SGLT-2mRNA in short antisense compounds and targeting mice and the rat is compared, and having more usefulness aspect the SGLT-2mRNA reducing, and onset is faster.This paper confirms that the short at interval aggressiveness (shortmer) of 2 ' MOE can improve usefulness and bioavailability than parent compound.

For example and only for purposes of illustration, the research of ISIS 370717 is disclosed, short antisense compounds is compared with longer parent significantly higher accumulation in renal tissue (about 50 milligrams/gram tissue).In addition, SGLT-2mRNA descends compared with the control above 80% (referring to embodiment 11).ISIS 370717 1-10-1 gapmer are used as the relevant oligonucleotide (sequence related oligos with varying motifs) of sequence that template for preparing has different die bodys.About the research that pterion, interval and total length around the assessment ISIS 370,717 12 mer oligonucleotide change can be referring to embodiment 12.Some evaluated die body comprises 1-10-1,2-8-2,1-8-1,3-6-3 and 1-6-1 (referring to the table 60 among the embodiment 12).Analyzed the effect of chemical compound to the SGLT2mRNA level.All die bodys all suppress SGLT2 expression in the body in the dose dependent mode.Find that 1-10-1,2-8-2 and 1-8-1 gapmer have usefulness especially.Use these die bodys, SGLT-2 mRNA descends compared with the control and surpasses 80%.

In certain embodiments, the invention provides targeting SGLT2 nucleic acid and have the short antisense compounds of the die body that is selected from 1-10-1 and 1-10-2 MOE gapmer.(referring to the table 62 among the embodiment 13).Analyzed of the effect of some this chemical compound to rat SGLT2mRNA.Presentation of results in the table 63,1-10-1 and 1-10-2 MOE gapmer all are to suppress SGLT2 expression in the body in the dose dependent mode, and the decline of SGLT-2mRNA can reach and surpass 80%.

Some other 1-10-1 and 2-8-2MOE gapmer (referring to for example embodiment 14 and 15) in mice and rat body inner model, have also been assessed.Many 1-10-1 and 2-8-2MOEgapmer have realized that under relatively low oligonucleotide concentration SGLT-2mRNA descends above 80%, and without any toxic action.

In another non-limiting example, also analyzed the effect of 388625 couples of Canis familiaris L. SGLT2mRNA of ISIS level.To the research explanation that Canis familiaris L. carries out, can realize that under the dosage of 1mg/kg/wk the expression of SGLT2 is suppressed above 80%.Under high slightly dosage, can realize bigger inhibition.Also confirm in Canis familiaris L., to give ISIS 388625 and improved glucose tolerance.In the glucose tolerance test of standard, to compare with saline control, the peak value plasma glucose levels on average descends and surpasses 50%, and the decline of glucose subsequently weakens (referring to embodiment 17).Equally, compare with the animal of PBS and control treatment, confirm in diabetes rat model, short antisense compounds is passed in time and can significantly be reduced plasma glucose levels and HbA1C (referring to embodiment 16).

Animal in all researchs all further carries out the toxicity assessment.For example, TBW and liver, spleen and kidney weight have been assessed.The significant change of spleen, liver weight or body weight can show that certain specific compound can cause toxic action.Find that all variations are all in the error span scope.In therapeutic process or when research stops, do not observe significant body weight change.Do not observe the significant change of liver or spleen weight.

The short antisense compounds of some targeting SGLT2 nucleic acid

In certain embodiments, lacking the antisense compounds targeting hasThe SGLT2 nucleic acid of the sequence of searching number NM_003041.1 (being incorporated herein) with SEQ ID NO:2.In some this embodiment, short antisense compounds and the SEQID NO:3 of targeting SEQ ID NO:3 have at least 90% complementarity.In some this embodiment, the short antisense compounds of targeting SEQ IDNO:3 and SEQ ID NO:3 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:1 of targeting SEQ ID NO:3 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:3 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 4 and 5.

The nucleotide sequence that each SEQ ID NO provides in the table 4 and 5 is independent of any modification to sugar moieties, Dan Julian key or nuclear base.Thus, the defined short antisense compounds of certain SEQ ID NO can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 4 and 5 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:3.Table 4 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:3.Table 5 has been enumerated the short antisense compounds that has one or two mispairing with respect to SEQ ID NO:3.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment flank of 10 2 '-deoxyribonucleotides is connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Table 4: the short antisense compounds of targeting SEQ ID NO:3

ISIS No	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						379684	84	95	TGTCAGCAGGAT	1-10-1 MOE	214
405193	113	124	CAGCAGGAAATA	2-8-2 MOE	215
						405194	114	125	CCAGCAGGAAAT	2-8-2 MOE	216
405195	115	126	ACCAGCAGGAAA	2-8-2 MOE	217
						405196	116	127	GACCAGCAGGAA	2-8-2 MOE	218
405197	117	128	TGACCAGCAGGA	2-8-2 MOE	219
						379685	117	128	TGACCAGCAGGA	1-10-1 MOE	219
405198	118	129	ATGACCAGCAGG	2-8-2 MOE	221
						405199	119	130	AATGACCAGCAG	2-8-2 MOE	222
405200	120	131	CAATGACCAGCA	2-8-2 MOE	223
						405201	121	132	CCAATGACCAGC	2-8-2 MOE	224
379686	135	146	ACCACAAGCCAA	1-10-1 MOE	225
						379711	172	183	TAGCCGCCCACA	1-10-1 MOE	226
388628	172	183	TAGCCGCCCACA	2-8-2 MOE	226
						405202	207	218	CCGGCCACCACA	2-8-2 MOE	228
405203	208	219	ACCGGCCACCAC	2-8-2 MOE	229
						405204	236	247	GATGTTGCTGGC	2-8-2 MOE	230
379687	236	247	GATGTTGCTGGC	1-10-1 MOE	230
						405205	237	248	CGATGTTGCTGG	2-8-2 MOE	232
405206	238	249	CCGATGTTGCTG	2-8-2 MOE	233
						405207	239	250	GCCGATGTTGCT	2-8-2 MOE	234
405208	240	251	TGCCGATGTTGC	2-8-2 MOE	235

405209	241	252	CTGCCGATGTTG	2-8-2 MOE	236
						405210	260	271	CAGGCCCACAAA	2-8-2 MOE	237
405211	261	272	CCAGGCCCACAA	2-8-2 MOE	238
						405212	262	273	GCCAGGCCCACA	2-8-2 MOE	239
379688	288	299	CCAAGCCACTTG	1-10-1 MOE	240
						379689	318	329	AGAGCGCATTCC	1-10-1 MOE	241
379690	435	446	ACAGGTAGAGGC	1-10-1 MOE	242
						405248	474	485	AGATCTTGGTGA	2-8-2 MOE	243
379691	474	485	AGATCTTGGTGA	1-10-1 MOE	243
						382676	527	539	TGTTCCAGCCCAG	1-10-2 MOE	245
388625	528	539	TGTTCCAGCCCA	2-8-2 MOE	246
						389780	528	539	TGTTCCAGCCCA	1-9-2 MOE	246
379692	528	539	TGTTCCAGCCCA	1-10-1 MOE	246
						392170	528	539	TGTTCCAGCCCA	1-10-1 methylene oxygen base BNA	246
392173	528	539	TGTTCCAGCCCA	2-8-2 methylene oxygen base BNA	246
						405213	529	540	ATGTTCCAGCCC	2-8-2 MOE	251
405214	564	575	TGGTGATGCCCA	2-8-2 MOE	252
						405215	565	576	ATGGTGATGCCC	2-8-2 MOE	253
405216	566	577	CATGGTGATGCC	2-8-2 MOE	254
						379693	566	577	CATGGTGATGCC	1-10-1 MOE	254
405217	567	578	TCATGGTGATGC	2-8-2 MOE	256
						405218	568	579	ATCATGGTGATG	2-8-2 MOE	257
405219	587	598	CCCTCCTGTCAC	2-8-2 MOE	258
						405220	588	599	GCCCTCCTGTCA	2-8-2 MOE	259
405221	589	600	AGCCCTCCTGTC	2-8-2 MOE	260
						405222	590	601	CAGCCCTCCTGT	2-8-2 MOE	261
405223	591	602	CCAGCCCTCCTG	2-8-2 MOE	262
						405224	592	603	GCCAGCCCTCCT	2-8-2 MOE	263
379694	629	640	GACGAAGGTCTG	1-10-1 MOE	264
						405225	707	718	GTATTTGTCGAA	2-8-2 MOE	265
379695	737	748	GGACACCGTCAG	1-10-1 MOE	266
						379696	974	985	CAGCTTCAGGTA	1-10-1 MOE	267
405226	998	1009	CATGACCATGAG	2-8-2 MOE	268
						405227	999	1010	GCATGACCATGA	2-8-2 MOE	269
405228	1000	1011	GGCATGACCATG	2-8-2 MOE	270
						405229	1001	1012	TGGCATGACCAT	2-8-2 MOE	271
405230	1002	1013	CTGGCATGACCA	2-8-2 MOE	272
						379697	1002	1013	CTGGCATGACCA	1-10-1 MOE	272
405231	1003	1014	CCTGGCATGACC	2-8-2 MOE	274
						379698	1091	1102	GCAGCCCACCTC	1-10-1 MOE	275
405232	1092	1103	AGCAGCCCACCT	2-8-2 MOE	276
						405233	1093	1104	GAGCAGCCCACC	2-8-2 MOE	277
405234	1130	1141	CATGAGCTTCAC	2-8-2 MOE	278
						405235	1131	1142	GCATGAGCTTCA	2-8-2 MOE	279
382677	1131	1143	GGCATGAGCTTCA	1-10-2 MOE	280
						388626	1132	1143	GGCATGAGCTTC	2-8-2 MOE	281
379699	1132	1143	GGCATGAGCTTC	1-10-1 MOE	281
						405236	1133	1144	GGGCATGAGCTT	2-8-2 MOE	283
405237	1157	1168	CAGCATGAGTCC	2-8-2 MOE	284

405238	1158	1169	CCAGCATGAGTC	2-8-2 MOE	285
						379700	1158	1169	CCAGCATGAGTC	1-10-1 MOE	285
405239	1159	1170	GCCAGCATGAGT	2-8-2 MOE	287
						379701	1230	1241	CCATGGTGAAGA	1-10-1 MOE	288
405240	1542	1553	CACAGCTGCCCG	2-8-2 MOE	289
						405241	1543	1554	ACACAGCTGCCC	2-8-2 MOE	290
405242	1544	1555	CACACAGCTGCC	2-8-2 MOE	291
						382678	1544	1556	GCACACAGCTGCC	1-10-2 MOE	292
388627	1545	1556	GCACACAGCTGC	2-8-2 MOE	293
						379702	1545	1556	GCACACAGCTGC	1-10-1 MOE	293
379703	1701	1712	GCCGGAGACTGA	1-10-1 MOE	295
						405243	1976	1987	ATTGAGGTTGAC	2-8-2 MOE	296
405244	1977	1988	CATTGAGGTTGA	2-8-2 MOE	297
						405245	1978	1989	GCATTGAGGTTG	2-8-2 MOE	298
405246	1979	1990	GGCATTGAGGTT	2-8-2 MOE	299
						405247	1980	1991	GGGCATTGAGGT	2-8-2 MOE	300

Table 5: targeting SEQ ID NO:3 and have the short antisense compounds of 1 or 2 mispairing

ISIS No	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						405200	96	107	CAATGACCAGCA	2-8-2 MOE	223
405215	382	393	ATGGTGATGCCC	2-8-2 MOE	253
						405216	383	394	CATGGTGATGCC	2-8-2 MOE	254
379693	383	394	CATGGTGATGCC	1-10-1 MOE	254
						379701	471	482	CCATGGTGAAGA	1-10-1 MOE	288
405218	472	483	ATCATGGTGATG	2-8-2 MOE	257
						405246	536	547	GGCATTGAGGTT	2-8-2 MOE	299
405248	570	581	AGATCTTGGTGA	2-8-2 MOE	243
						379691	570	581	AGATCTTGGTGA	1-10-1 MOE	243
379698	683	694	GCAGCCCACCTC	1-10-1 MOE	275
						405232	684	695	AGCAGCCCACCT	2-8-2 MOE	276
379711	685	696	TAGCCGCCCACA	1-10-1 MOE	226
						388628	685	696	TAGCCGCCCACA	2-8-2 MOE	226
379698	950	961	GCAGCCCACCTC	1-10-1 MOE	275
						405232	951	962	AGCAGCCCACCT	2-8-2 MOE	276
405235	978	989	GCATGAGCTTCA	2-8-2 MOE	279
						382677	978	990	GGCATGAGCTTCA	1-10-2 MOE	280
388626	979	990	GGCATGAGCTTC	2-8-2 MOE	281
						379699	979	990	GGCATGAGCTTC	1-10-1 MOE	281
405236	980	991	GGGCATGAGCTT	2-8-2 MOE	283
						379698	1043	1054	GCAGCCCACCTC	1-10-1 MOE	275
405239	1171	1182	GCCAGCATGAGT	2-8-2 MOE	287
						405209	1213	1224	CTGCCGATGTTG	2-8-2 MOE	236
405233	1364	1375	GAGCAGCCCACC	2-8-2 MOE	277
						405240	1366	1377	CACAGCTGCCCG	2-8-2 MOE	289
405211	1500	1511	CCAGGCCCACAA	2-8-2 MOE	238
						405212	1501	1512	GCCAGGCCCACA	2-8-2 MOE	239
379695	1643	1654	GGACACCGTCAG	1-10-1 MOE	266

379698	1875	1886	GCAGCCCACCTC	1-10-1 MOE	275
						405239	1993	2004	GCCAGCATGAGT	2-8-2 MOE	287
405211	2210	2221	CCAGGCCCACAA	2-8-2 MOE	238
						405212	2211	2222	GCCAGGCCCACA	2-8-2 MOE	239

In certain embodiments, the target zone is the nucleotide 85-184 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 85-184 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 85-184 comprises the nucleotide sequence that is selected from SEQ ID NO 214,215,216,217,218,219,221,222,223,224,225 or 227.In some this embodiment, the short antisense compounds of the nucleotide 85-184 of targeting SEQ ID NO:3 is selected from Isis No 379684,405193,405194,405195,405196,405197,379685,405198,405199,405200,405201,379686,379711 or 388628.

In certain embodiments, the target zone is the nucleotide 113-132 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 113-132 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 113-132 comprises the nucleotide sequence that is selected from SEQ ID NO 215,216,217,218,219,221,222,223 or 224.In some this embodiment, the short antisense compounds of the nucleotide 113-132 of targeting SEQ ID NO:3 is selected from Isis No 405193,405194,405195,405196,405197,379685,405198,405199,405200 or 405201.

In certain embodiments, the target zone is the nucleotide 207-329 of SEQ ID NO:3.3. lack in certain embodiments, the nucleotide 207-329 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 207-329 comprises the nucleotide sequence that is selected from SEQ ID NO 228,229,230,232,233,234,235,236,237,238,239,240 or 241.In some this embodiment, the short antisense compounds of the nucleotide 207-329 of targeting SEQ ID NO:3 is selected from Isis No405202,405203,405204,379687,405205,405206,405207,405208,405209,405210,405211,405212,379688 or 379689.

In certain embodiments, the target zone is the nucleotide 207-273 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 207-273 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 207-273 comprises the nucleotide sequence that is selected from SEQ IDNO 228,229,230,232,233,234,235,236,237,238 or 239.In some this embodiment, the short antisense compounds of the nucleotide 207-273 of targeting SEQ IDNO:3 is selected from Isis No 405202,405203,405204,379687,405205,405206,405207,405208,405209,405210,405211 or 405212.

In certain embodiments, the target zone is the nucleotide 207-219 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 207-219 of antisense compounds targeting SEQ ID NO:3.3. in some this embodiment, the short antisense compounds of targeted nucleotide 207-219 comprises the nucleotide sequence that is selected from SEQ ID NO 228 or 229.In some this embodiment, the short antisense compounds of the nucleotide 207-219 of targeting SEQ ID NO:3 is selected from Isis NO..405202 or 405203.

In certain embodiments, the target zone is the nucleotide 236-252 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 236-252 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 236-252 comprises the nucleotide sequence that is selected from SEQ ID NO 230,232,233,234,235 or 236.In some this embodiment, the short antisense compounds of the nucleotide 236-252 of targeting SEQ ID NO:3 is selected from Isis NO.405204,379687,405205,405206,405207,405208 or 405209.

In certain embodiments, the target zone is the nucleotide 260-273 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 260-273 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 260-273 comprises the nucleotide sequence that is selected from SEQ ID NO 237,238 or 239.In some this embodiment, the short antisense compounds of the nucleotide 260-273 of targeting SEQ ID NO:3 is selected from Isis NO.405210,405211 or 405212.

In certain embodiments, the target zone is the nucleotide 435-640 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 435-640 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 435-640 comprises the nucleotide sequence that is selected from SEQ ID NO 242,243,245,246,251,252,253,254,256,257,258,259,260,261,262,263 or 264.In some this embodiment, the short antisense compounds of the nucleotide 435-640 of targeting SEQ ID NO:3 is selected from Isis NO.379690,405248,379691,389780,379692,382676,388625,392170,392173,405213,405214,405215,405216,379693,405217,405218,405219,405220,405221,405222,405223,405224 or 379694.

In certain embodiments, the target zone is the nucleotide 527-540 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 527-540 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 527-540 comprises the nucleotide sequence that is selected from SEQ ID NO 245,246 or 251.In some this embodiment, the short antisense compounds of the nucleotide 527-540 of targeting SEQ ID NO:3 is selected from Isis NO.389780,379692,382676,388626,392170,392173 or 405213.

In certain embodiments, the target zone is the nucleotide 564-603 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 564-603 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 564-603 comprises the nucleotide sequence that is selected from SEQ ID NO 252,253,254,256,257,258,259,260,261,262 or 263.In some this embodiment, the short antisense compounds of the nucleotide 564-603 of targeting SEQ IDNO:3 is selected from Isis NO.405214,405215,405216,379693,405217,405218,405219,405220,405221,405222,405223 or 405224.

In certain embodiments, the target zone is the nucleotide 564-579 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 564-579 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 564-579 comprises the nucleotide sequence that is selected from SEQ ID NO 252,253,254,256 or 257.In some this embodiment, the short antisense compounds of the nucleotide 564-579 of targeting SEQ ID NO:3 is selected from Isis NO.405214,405215,405216,379693,405217 or 405218.

In certain embodiments, the target zone is the nucleotide 587-603 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 587-603 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 587-603 comprises the nucleotide sequence that is selected from SEQ ID NO 258,259,260,261,262 or 263.In some this embodiment, the short antisense compounds of the nucleotide 587-603 of targeting SEQ ID NO:3 is selected from Isis NO.405219,405220,405221,405222,405223 or 405224.

In certain embodiments, the target zone is the nucleotide 974-1014 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 974-1014 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 974-1014 comprises the nucleotide sequence that is selected from SEQ ID NO 267,268,269,270,271,272 or 274.In some this embodiment, the short antisense compounds of the nucleotide 974-1014 of targeting SEQ ID NO:3 is selected from Isis NO.379696,405226,405227,405228,405229,405230,379697 or 405231.

In certain embodiments, the target zone is the nucleotide 998-1014 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 998-1014 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 998-1014 comprises the nucleotide sequence that is selected from SEQ ID NO 268,269,270,271,272 or 274.In some this embodiment, the short antisense compounds of the nucleotide 998-1014 of targeting SEQ ID NO:3 is selected from Isis NO.405226,405227,405228,405229,405230,379697 or 405231.

In certain embodiments, the target zone is the nucleotide 1091-1170 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 1091-1170 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 1091-1170 comprises the nucleotide sequence that is selected from SEQ ID NO 275,276,277,278,279,280,281,283,284,285,286 or 287.In some this embodiment, the short antisense compounds of the nucleotide 1091-1170 of targeting SEQ ID NO:3 is selected from Isis NO.379698,405232,405233,405234,405235,388626,379699,382677,405236,405237,405238,379700 or 405239.

In certain embodiments, the target zone is the nucleotide 1091-1104 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 1091-1104 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 1091-1104 comprises the nucleotide sequence that is selected from SEQ ID NO 275,276 or 277.In some this embodiment, the short antisense compounds of the nucleotide 1091-1104 of targeting SEQ ID NO:3 is selected from Isis NO.379698,405232 or 405233.

In certain embodiments, the target zone is the nucleotide 1130-1144 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 1130-1144 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 1130-1144 comprises the nucleotide sequence that is selected from SEQ ID NO 278,279,280,281 or 283.In some this embodiment, the short antisense compounds of the nucleotide 1130-1144 of targeting SEQ ID NO:3 is selected from Isis NO.405234,405235,388626,379699,382677 or 405236.

In certain embodiments, the target zone is the nucleotide 1157-1170 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 1157-1170 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 1157-1170 comprises the nucleotide sequence that is selected from SEQ ID NO 284,285 or 287.In some this embodiment, the short antisense compounds of the nucleotide 1157-1170 of targeting SEQ ID NO:3 is selected from Isis NO.405237,405238,379700 or 405239.

In certain embodiments, the target zone is the nucleotide 1542-1556 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 1542-1556 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 1542-1556 comprises the nucleotide sequence that is selected from SEQ ID NO 289,290,291,292 or 293.In some this embodiment, the short antisense compounds of the nucleotide 1542-1556 of targeting SEQ ID NO:3 is selected from Isis NO.405240,405241,405242,388629,379702 or 382678.

In certain embodiments, the target zone is the nucleotide 1976-1991 of SEQ ID NO:3.In certain embodiments, lack the nucleotide 1976-1991 of antisense compounds targeting SEQ ID NO:3.In some this embodiment, the short antisense compounds of targeted nucleotide 1976-1991 comprises the nucleotide sequence that is selected from SEQ ID NO 296,297,298,299 or 300.In some this embodiment, the short antisense compounds of the nucleotide 1976-1991 of targeting SEQ ID NO:3 is selected from Isis NO.405243,405244,405245,405246 or 405247.

In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in one or more pterions of the short gapmer of targeting SGLT2 nucleic acid in chemical compound and modify.In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting SGLT2 nucleic acid is 16 monomers.

In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting SGLT2 nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating SGLT2.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting SGLT2 nucleic acid, and wherein the short antisense compounds of this targeting SGLT2 nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting SGLT2 nucleic acid is regulated the expression of SGLT2.

In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 8 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 9 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 10 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 11 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 12 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 13 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 14 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 15 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for adjusting SGLT2 comprises that using length is the short antisense compounds of 16 monomeric targeting SGLT2 nucleic acid.

In certain embodiments, the method for the expression of adjusting SGLT2 comprises that use comprises the short antisense compounds of 9-15 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for the expression of adjusting SGLT2 comprises that use comprises the short antisense compounds of 10-15 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for the expression of adjusting SGLT2 comprises that use comprises the short antisense compounds of 12-14 monomeric targeting SGLT2 nucleic acid.In certain embodiments, the method for the expression of adjusting SGLT2 comprises the short antisense compounds that uses the targeting SGLT2 nucleic acid that comprises 12-14 nucleotide or nucleoside.

3.PCSK9

To suffer from the high LDL-C level in the individuality of autosomal dominant hypercholesterolemia (ADH), with the coding LDL-receptor (LDL-R), apolipoprotein B (apoB) or crude protein (proprotein) invertase subtilisin/kexin type 9 (PCSK9) gene in sudden change associate (Abifadel et al., Nat.Genet., 2003,34:154-156).Obtain function mutation (gain-of-function mutation) when relevant with high LDL-C level in finding PCSK9, PCSK9 is accredited as three locus relevant with ADH.The interior assembling of born of the same parents and the secretion of the lipoprotein of triglyceride are rich in the ApoB participation, and are the parts of LDL-R.PCSK9 is considered to reduce the LDL-R expression in the liver.The liver picked-up that contains the ApoB lipoprotein during the reduction that LDL-R expresses causes circulating reduces, and this causes cholesterol to raise.

Definition

" PCSK9 " is that its expression is treated by lacking gene outcome or the protein that antisense compounds is regulated.

" PCSK9 nucleic acid " means the nucleic acid of any coding PCSK9.For example, in certain embodiments, PCSK9 nucleic acid include but not limited to the to encode DNA sequence of PCSK9, RNA sequence of transcribing from the DNA of coding PCSK9 and the mRNA sequence of coding PCSK9.

" PCSK9mRNA " means the mRNA of coding PCSK9.

PCSK9 treats indication

In certain embodiments, the invention provides the method for the expression of regulating the PCSK9 in the individuality, described method comprises the short antisense compounds that gives targeting PCSK9 nucleic acid.In certain embodiments, the invention provides the individual method of treatment, described method comprises and gives one or more pharmaceutical compositions of the present invention.In certain embodiments, individuality suffers from hypercholesterolemia, Combination dyslipidemia, atherosclerosis, the atherosclerotic risk of development, coronary heart disease, coronary disease medical history, the coronary heart disease of early showing effect, one or more coronary heart disease risk factors, type ii diabetes, type ii diabetes companion dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipemia, high fatty acid mass formed by blood stasis, fatty degeneration of liver, non-alcoholic stellato-hepatitis or non-alcohol fatty liver.

The guilding principle that lipid reduces therapy by u.s. national cholesterol education program (NCEP) adult treatment expert scheme III (ATPIII) in calendar year 2001 set up and upgraded in 2004 (Grundyet al., Circulation, 2004,110,227-239).This guilding principle comprises acquisition complete lipoprotein general picture (profile) after fasting in 9-12 hour usually, to determine LDL-C, T-CHOL and HDL-C level.According to the guilding principle of up-to-date formulation, 130-159mg/dL, 160-189mg/dL and think critical height, height and high level respectively more than or equal to the LDL-C level of 190mg/dL.200-239mg/dL and think critical height and high level respectively more than or equal to the total cholesterol level of 240mg/dL.HDL-C level less than 40mg/dL is thought low-level.

In certain embodiments, this individuality has been confirmed as carrying out lipid reduction therapy.In some this embodiment, according to upgrading (Grundyet al. in calendar year 2001 foundation and in 2004 by u.s. national cholesterol education program (NCEP) adult treatment expert scheme III (ATPIII), Circulation, 2004,110, guilding principle 227-239), this individuality have been confirmed as carrying out lipid and have reduced therapy.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 190mg/dL.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 160mg/dL.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 130mg/dL.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 100mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 160mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 130mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 100mg/dL.In some this embodiment, this individuality should keep LDL-C to be lower than 70mg/dL.

In certain embodiments, the invention provides the method that reduces the ApoB in the individuality.In certain embodiments, the invention provides the method that contains the ApoB lipoprotein that reduces in the individuality.In certain embodiments, the invention provides the method that reduces the LDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the VLDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the IDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the non-IDL-C in the individuality.In certain embodiments, the invention provides the method that reduces the Lp (a) in the individuality.In certain embodiments, the invention provides the method that reduces the serum triglycerides in the individuality.In certain embodiments, the invention provides the method that reduces the liver tg in the individuality.In certain embodiments, the invention provides the method that reduces the Ox-LDL-C in the individuality.In certain embodiments, the invention provides the particulate method of little LDL that reduces in the individuality.In certain embodiments, the invention provides the particulate method of little VLDL that reduces in the individuality.In certain embodiments, the invention provides the method that reduces the phospholipid in the individuality.In certain embodiments, the invention provides the method that reduces the oxidized phospholipids in the individuality.

In certain embodiments, method provided by the invention does not reduce HDL-C.In certain embodiments, method provided by the invention does not cause the accumulation of lipid in liver.

In certain embodiments, the pharmaceutical composition that comprises the short antisense compounds of targeting PCSK9 nucleic acid is for using in treatment.In certain embodiments, described treatment is the reduction of LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp (a), serum triglycerides, liver tg, Ox-LDL-C, little LDL granule, little VLDL, phospholipid or oxidized phospholipids in the individuality.In certain embodiments, described treatment is to following treatment of diseases: hypercholesterolemia, Combination dyslipidemia, atherosclerosis, the atherosclerotic risk of development, coronary heart disease, coronary disease medical history, the coronary heart disease of early showing effect, one or more coronary heart disease risk factors, type ii diabetes, type ii diabetes companion dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipemia, high fatty acid mass formed by blood stasis, fatty degeneration of liver, non-alcoholic stellato-hepatitis or non-alcohol fatty liver.In other embodiments, treatment is the reduction of CHD risk.In certain embodiments, treatment is atherosclerotic prevention.In certain embodiments, treatment is the prevention of coronary heart disease.

In certain embodiments, the pharmaceutical composition that will comprise the short antisense compounds of targeting PCSK9 nucleic acid is used for preparing the medicine in order to LDL-C, ApoB, VLDL-C, IDL-C, non-HDL-C, Lp (a), serum triglycerides, liver tg, Ox-LDL-C, little LDL granule, little VLDL, phospholipid or the oxidized phospholipids that reduces individuality.In certain embodiments, the pharmaceutical composition that will comprise the short antisense compounds of targeting PCKS9 nucleic acid is used to prepare in order to reduce the medicine of coronary heart disease risk.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid is used to prepare in order to treat the medicine of following disease: hypercholesterolemia, Combination dyslipidemia, atherosclerosis, the atherosclerotic risk of development, coronary heart disease, coronary disease medical history, the coronary heart disease of early showing effect, one or more coronary heart disease risk factors, type ii diabetes, type ii diabetes companion dyslipidemia, dyslipidemia, hypertriglyceridemia, hyperlipemia, high fatty acid mass formed by blood stasis, fatty degeneration of liver, non-alcoholic stellato-hepatitis or non-alcohol fatty liver.

The PCSK9 combination treatment

In certain embodiments, one or more pharmaceutical compositions of the present invention are given altogether with one or more other medicament.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease identical with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease different with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament is designed to treat the ill effect of one or more pharmaceutical compositions of the present invention.In certain embodiments, one or more pharmaceutical compositions of the present invention are given altogether with another medicament, to treat the ill effect of this another medicament.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given simultaneously.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given at different time.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is prepared together in single dosage form.In certain embodiments, with other separately preparation of medicament of one or more pharmaceutical compositions of the present invention and one or more.

In certain embodiments, can comprise that lipid reduces medicament with the medicament that pharmaceutical composition of the present invention gives altogether.In some this embodiment, can include but not limited to atorvastatin, simvastatin, rosuvastatin with the medicament that pharmaceutical composition of the present invention gives altogether and according to Ezetimibe.In some this embodiment, lipid is reduced medicament give before the pharmaceutical composition of the present invention giving.In some this embodiment, lipid is reduced medicament after giving pharmaceutical composition of the present invention, give.In some this embodiment, lipid is reduced medicament when giving pharmaceutical composition of the present invention, give.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is identical with the dosage that gives separately to be given when lipid reduces medicament.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is lower than the dosage that is given when giving lipid reduction medicament separately.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is higher than the dosage that is given when giving lipid reduction medicament separately.

In certain embodiments, it is the HMG-CoA reductase inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, the HMG-CoA reductase inhibitor is an inhibin.In some this embodiment, inhibin is selected from atorvastatin, simvastatin, pravastatin, fluvastatin and rosuvastatin.

In certain embodiments, it is cholesterol absorption inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, cholesterol absorption inhibitor is according to Ezetimibe.

In certain embodiments, it is HMG-CoA reductase inhibitor and cholesterol absorption inhibitor formulated together altogether that the lipid that gives altogether reduces medicament.In some this embodiment, it is according to Ezetimibe/simvastatin that lipid formulated together altogether reduces medicament.

In certain embodiments, it is microsomal triglyceride transfer protein inhibitor (MTP inhibitor) that the lipid that gives altogether reduces medicament.

In certain embodiments, the medicament that gives altogether is the oligonucleotide of targeting ApoB nucleic acid.

In certain embodiments, the medicament that gives altogether is a bile acid chelating agent.In some this embodiment, bile acid chelating agent is selected from colestyramine, colestipol and colesevelam.

In certain embodiments, the medicament that gives altogether is a nicotinic acid.In some this embodiment, nicotinic acid is selected from rapid release nicotinic acid, extended release nicotinic acid and slow release nicotinic acid.

In certain embodiments, the medicament that gives altogether is a Carboxymethylcellulose.In some this embodiment, Carboxymethylcellulose is selected from gemfibrozil, fenofibrate, chlorine Bei Te, bezafibrate and ciprofibrate.

More examples of the medicament that can give altogether with pharmaceutical composition of the present invention include but not limited to corticosteroid (including but not limited to prednisone); Immunoglobulin (includes but not limited to vein immunoglobulin (IVIg); Analgesic (for example acetaminophen); Antiinflammatory (including but not limited to NSAID (non-steroidal anti-inflammatory drug) (for example ibuprofen, COX-1 inhibitor and cox 2 inhibitor)); Salicylate; Antibiotic; Antiviral agent; Antifungal; Antidiabetic medicine (for example biguanide, glucosidase inhibitor, insulin, sulfonylureas and thiazolidinediones); The adrenergic modulation agent; Diuretic; Hormone (for example short anabolism steroid, androgen, estrogen, calcitonin, progesterone, somatostatin and thyroxin); Immunomodulator; Muscle relaxant; Hydryllin; Osteoporosis medicament (for example diphosphonate, calcitonin and estrogen); Prostaglandin; Antineoplastic agent; The spiritual healing medicament; Tranquilizer; Poison Oak Tree (oak) or black poison wood (sumac) product; Antibody and vaccine.

In certain embodiments, pharmaceutical composition of the present invention can be reduced therapy in conjunction with lipid gives.In some this embodiment, it is that curative life style changes that lipid reduces therapy.In some this embodiment, it is the LDL Apheresis that lipid reduces therapy.

The short antisense compounds of some targeting PCSK9 nucleic acid

In certain embodiments, lacking the antisense compounds targeting has

The PCSK9 nucleic acid of the sequence of searching number NM_174936.2 (being incorporated herein) with SEQ ID NO:4.

In some this embodiment, short antisense compounds and the SEQID NO:4 of targeting SEQ ID NO:4 have at least 90% complementarity.In some this embodiment, the short antisense compounds of targeting SEQ IDNO:4 and SEQ ID NO:4 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:4 of targeting SEQ ID NO:4 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 6 and 7.

The nucleotide sequence that each SEQ ID NO provides in the table 6 and 7 is independent of any to connecting the modification of key or nuclear base between sugar moieties, nucleoside.Thus, the defined short antisense compounds of certain SEQ ID NO can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 6 and 7 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:4.Table 6 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:4.Table 7 has been enumerated the short antisense compounds that has one or two mispairing with respect to SEQ ID NO:4.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Table 6: the short antisense compounds of targeting SEQ ID NO:4

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						400297	695	708	ATGGGGCAACTTCA	2-10-2 MOE	329
400298	696	709	CATGGGGCAACTTC	2-10-2 MOE	330
						400299	697	710	ACATGGGGCAACTT	2-10-2 MOE	331
400300	742	755	GGGATGCTCTGGGC	2-10-2 MOE	332
						400301	757	770	CGCTCCAGGTTCCA	2-10-2 MOE	333
400302	828	841	GATACACCTCCACC	2-10-2 MOE	334
						400303	829	842	AGATACACCTCCAC	2-10-2 MOE	335
400304	830	843	GAGATACACCTCCA	2-10-2 MOE	336
						400305	937	950	GCCTGTCTGTGGAA	2-10-2 MOE	337
400306	952	965	CTGTCACACTTGCT	2-10-2 MOE	338
						400307	988	1001	CGGCCGCTGACCAC	2-10-2 MOE	339
400308	989	1002	CCGGCCGCTGACCA	2-10-2 MOE	340
						400309	990	1003	CCCGGCCGCTGACC	2-10-2 MOE	341
400310	991	1004	TCCCGGCCGCTGAC	2-10-2 MOE	342
						400311	992	1005	ATCCCGGCCGCTGA	2-10-2 MOE	343
400312	993	1006	CATCCCGGCCGCTG	2-10-2 MOE	344
						400313	994	1007	GCATCCCGGCCGCT	2-10-2 MOE	345
400314	1057	1070	GTGCCCTTCCCTTG	2-10-2 MOE	346
						400315	1075	1088	ATGAGGGTGCCGCT	2-10-2 MOE	347
400316	1076	1089	TATGAGGGTGCCGC	2-10-2 MOE	348
						400317	1077	1090	CTATGAGGGTGCCG	2-10-2 MOE	349
400318	1078	1091	CCTATGAGGGTGCC	2-10-2 MOE	350
						400319	1093	1106	CGAATAAACTCCAG	2-10-2 MOE	351
400320	1094	1107	CCGAATAAACTCCA	2-10-2 MOE	352
						400321	1095	1108	TCCGAATAAACTCC	2-10-2 MOE	353
400322	1096	1109	TTCCGAATAAACTC	2-10-2 MOE	354
						400323	1147	1160	GCCAGGGGCAGCAG	2-10-2 MOE	355
400324	1255	1268	GAGTAGAGGCAGGC	2-10-2 MOE	356
						400325	1334	1347	CCCCAAAGTCCCCA	2-10-2 MOE	357
400326	1335	1348	TCCCCAAAGTCCCC	2-10-2 MOE	358
						400327	1336	1349	GTCCCCAAAGTCCC	2-10-2 MOE	359
400328	1453	1466	ACGTGGGCAGCAGC	2-10-2 MOE	360
						400329	1454	1467	CACGTGGGCAGCAG	2-10-2 MOE	361
400330	1455	1468	CCACGTGGGCAGCA	2-10-2 MOE	362
						400331	1456	1469	GCCACGTGGGCAGC	2-10-2 MOE	363
400332	1569	1582	CAGGGAACCAGGCC	2-10-2 MOE	364
						400333	1570	1583	TCAGGGAACCAGGC	2-10-2 MOE	365
400334	1571	1584	CTCAGGGAACCAGG	2-10-2 MOE	366
						400335	1572	1585	CCTCAGGGAACCAG	2-10-2 MOE	367
400336	1573	1586	TCCTCAGGGAACCA	2-10-2 MOE	368
						400337	1574	1587	GTCCTCAGGGAACC	2-10-2 MOE	369
400338	1575	1588	GGTCCTCAGGGAAC	2-10-2 MOE	370
						400339	1576	1589	TGGTCCTCAGGGAA	2-10-2 MOE	371
400340	1577	1590	CTGGTCCTCAGGGA	2-10-2 MOE	372
						400341	1578	1591	GCTGGTCCTCAGGG	2-10-2 MOE	373
400342	1621	1634	GTGCTGGGGGGCAG	2-10-2 MOE	374
						400343	1622	1635	GGTGCTGGGGGGCA	2-10-2 MOE	375

400344	1623	1636	GGGTGCTGGGGGGC	2-10-2 MOE	376
						400345	1624	1637	TGGGTGCTGGGGGG	2-10-2 MOE	377
400346	1738	1751	GAGCAGCTCAGCAG	2-10-2 MOE	378
						400347	1739	1752	GGAGCAGCTCAGCA	2-10-2 MOE	379
400348	1740	1753	TGGAGCAGCTCAGC	2-10-2 MOE	380
						400349	1741	1754	CTGGAGCAGCTCAG	2-10-2 MOE	381
400350	1834	1847	CCCTCACCCCCAAA	2-10-2 MOE	382
						400351	1835	1848	ACCCTCACCCCCAA	2-10-2 MOE	383
400352	1836	1849	CACCCTCACCCCCA	2-10-2 MOE	384
						400353	1837	1850	ACACCCTCACCCCC	2-10-2 MOE	385
400354	1838	1851	GACACCCTCACCCC	2-10-2 MOE	386
						400355	1839	1852	AGACACCCTCACCC	2-10-2 MOE	387
400356	1840	1853	TAGACACCCTCACC	2-10-2 MOE	388
						400357	2083	2096	TGGCAGCAGGAAGC	2-10-2 MOE	389
400358	2084	2097	ATGGCAGCAGGAAG	2-10-2 MOE	390
						400359	2085	2098	CATGGCAGCAGGAA	2-10-2 MOE	391
400360	2086	2099	GCATGGCAGCAGGA	2-10-2 MOE	392
						400361	2316	2329	GGCAGCAGATGGCA	2-10-2 MOE	393
400362	2317	2330	CGGCAGCAGATGGC	2-10-2 MOE	394
						400363	2318	2331	CCGGCAGCAGATGG	2-10-2 MOE	395
400364	2319	2332	TCCGGCAGCAGATG	2-10-2 MOE	396
						400365	2320	2333	CTCCGGCAGCAGAT	2-10-2 MOE	397
400366	2321	2334	GCTCCGGCAGCAGA	2-10-2 MOE	398
						400367	2322	2335	GGCTCCGGCAGCAG	2-10-2 MOE	399
400368	2323	2336	CGGCTCCGGCAGCA	2-10-2 MOE	400
						400369	2324	2337	CCGGCTCCGGCAGC	2-10-2 MOE	401
400370	2325	2338	GCCGGCTCCGGCAG	2-10-2 MOE	402
						400371	3543	3556	AGTTACAAAAGCAA	2-10-2 MOE	403
403739	988	1001	CGGCCGCTGACCAC	2-10-2 (6 ' S)-6 '-methyl-methylene oxygen base BNA	339
						403740	1455	1468	CCACGTGGGCAGCA	2-10-2 (6 ' S)-6 '-methyl-methylene oxygen base BNA	362

Table 7: targeting SEQ ID NO:4 and have the short antisense compounds of 1 or 2 mispairing

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						400323	349	362	GCCAGGGGCAGCAG	2-10-2 MOE	355
400370	679	692	GCCGGCTCCGGCAG	2-10-2 MOE	402
						400361	1860	1873	GGCAGCAGATGGCA	2-10-2 MOE	393
400323	1873	1886	GCCAGGGGCAGCAG	2-10-2 MOE	355
						400310	2257	2270	TCCCGGCCGCTGAC	2-10-2 MOE	342
400361	2653	2666	GGCAGCAGATGGCA	2-10-2 MOE	393
						400350	2811	2824	CCCTCACCCCCAAA	2-10-2 MOE	382
400351	2812	2825	ACCCTCACCCCCAA	2-10-2 MOE	383
						400352	2813	2826	CACCCTCACCCCCA	2-10-2 MOE	384
400353	2814	2827	ACACCCTCACCCCC	2-10-2 MOE	385
						400334	2966	2979	CTCAGGGAACCAGG	2-10-2 MOE	366

400332	3379	3392	CAGGGAACCAGGCC	2-10-2 MOE	364
						400340	3448	3461	CTGGTCCTCAGGGA	2-10-2 MOE	372
400341	3449	3462	GCTGGTCCTCAGGG	2-10-2 MOE	373

In certain embodiments, the target zone is the nucleotide 695-710 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 695-710 of targeting SEQ ID NO:4 comprises and is selected from SEQ ID NO:329,330 or 331 nucleotide sequence.In some this embodiment, the short antisense compounds of the nucleotide 695-710 of targeting SEQ ID NO:4 is selected from Isis NO.400297,400298 or 400299.

In certain embodiments, the target zone is the nucleotide 742-770 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 742-770 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 332 or 333.In some this embodiment, the short antisense compounds of the nucleotide 742-770 of targeting SEQ ID NO:4 is selected from Isis NO.400300 or 400301.

In certain embodiments, the target zone is the nucleotide 828-843 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 828-843 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 334,335 or 336.In some this embodiment, the short antisense compounds of the nucleotide 828-843 of targeting SEQ ID NO:4 is selected from ISIS No.400302,400303 or 400304.

In certain embodiments, the target zone is the nucleotide 937-1007 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 937-1007 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 337,338,339,340,341,342,343,344 or 345.In some this embodiment, the short antisense compounds of the nucleotide 937-1007 of targeting SEQ ID NO:4 is selected from Isis NO.400305,400306,400307,400308,400309,400310,400311,400312,400313 or 403739.

In certain embodiments, the target zone is the nucleotide 937-965 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 937-965 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 337 or 338.In some this embodiment, the short antisense compounds of the nucleotide 937-965 of targeting SEQ ID NO:4 is selected from Isis NO.400305 or 400306.

In certain embodiments, the target zone is the nucleotide 988-1007 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 988-1007 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 339,340,341,342,343,344 or 345.In some this embodiment, the short antisense compounds of the nucleotide 937-1007 of targeting SEQ IDNO:4 is selected from Isis NO.400307,400308,400309,400310,400311,400312,4003313 or 403739.

In certain embodiments, the target zone is the nucleotide 1057-1160 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1057-1160 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 346,347,348,349,350,351,352,353,354 or 355.In some this embodiment, the short antisense compounds of the nucleotide 1057-1160 of targeting SEQ ID NO:4 is selected from ISIS NO.400314,400315,400316,400317,400318,400319,400320,400321,400322 or 400323.

In certain embodiments, the target zone is the nucleotide 1057-1109 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1057-1109 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 346,347,348,349,350,351,352,353 or 354.In some this embodiment, the short antisense compounds of the nucleotide 1057-1109 of targeting SEQ ID NO:4 is selected from ISIS NO.400314,400315,400316,400317,400318,400319,400320,400321 or 400322.

In certain embodiments, the target zone is the nucleotide 1057-1091 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1057-1091 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 346,347,348,349 or 350.In some this embodiment, the short antisense compounds of the nucleotide 1057-1091 of targeting SEQ ID NO:4 is selected from ISIS NO.400314,400315,400316,400317 or 400318.

In certain embodiments, the target zone is the nucleotide 1093-1109 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1093-1109 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 351,352,353 or 354.In some this embodiment, the short antisense compounds of the nucleotide 1057-1109 of targeting SEQ ID NO:4 is selected from ISIS NO.400319,400320,400321 or 400322.

In certain embodiments, the target zone is the nucleotide 1334-1349 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1334-1349 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 357,358 or 359.In some this embodiment, the short antisense compounds of the nucleotide 1334-1349 of targeting SEQ ID NO:4 is selected from ISIS NO 400325,400326 or 400327.

In certain embodiments, the target zone is the nucleotide 1453-1469 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1453-1469 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 360,361,362 or 363.In some this embodiment, the short antisense compounds of the nucleotide 1453-1469 of targeting SEQ ID NO:4 is selected from ISIS NO 400328,400329,400330,400331 or 403470.

In certain embodiments, the target zone is the nucleotide 1569-1591 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1569-1591 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 364,365,366,367,368,369,370,371,372 or 373.In some this embodiment, the short antisense compounds of the nucleotide 1569-1591 of targeting SEQ ID NO:4 is selected from ISIS NO 400332,400333,400334,400335,400336,400337,400338,400339,400340 or 400341.

In certain embodiments, the target zone is the nucleotide 1621-1637 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1621-1637 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 374,375,376 or 377.In some this embodiment, the short antisense compounds of the nucleotide 1621-1637 of targeting SEQ ID NO:4 is selected from ISIS NO 400342,400343,400344 or 400345.

In certain embodiments, the target zone is the nucleotide 1738-1754 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1738-1754 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 378,379,380 or 381.In some this embodiment, the short antisense compounds of the nucleotide 1738-1754 of targeting SEQ ID NO:4 is selected from ISIS NO 400346,400347,400348 or 400349.

In certain embodiments, the target zone is the nucleotide 1834-1853 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 1834-1853 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 382,383,384,385,386,387 or 388.In some this embodiment, the short antisense compounds of the nucleotide 1834-1853 of targeting SEQ IDNO:4 is selected from ISIS NO 400350,400351,400352,400353,400354,400355 or 400356.

In certain embodiments, the target zone is the nucleotide 2083-2099 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 2083-2099 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 389,390,391 or 392.In some this embodiment, the short antisense compounds of the nucleotide 2083-2099 of targeting SEQ ID NO:4 is selected from ISIS NO400357,400358,400359 or 400360.

In certain embodiments, the target zone is the nucleotide 2316-2338 of SEQ ID NO:4.In some this embodiment, the short antisense compounds of the nucleotide 2316-2338 of targeting SEQ ID NO:4 comprises the nucleotide sequence that is selected from SEQ ID NO 393,394,395,396,397,398,399,400,401 or 402.In some this embodiment, the short antisense compounds of the nucleotide 2316-2338 of targeting SEQ ID NO:4 is selected from ISIS NO 400361,400362,400363,400364,400365,400366,400367,400368,400369 or 400370.

In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting PCSK9 nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid can have short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3 and 1-9-2.

In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting PCSK9 nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting PCSK9 nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating PCSK9.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting PCSK9 nucleic acid, and wherein the short antisense compounds of this targeting PCSK9 nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting PCSK9 nucleic acid is regulated the expression of PCSK9.

In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 8 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 9 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 10 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 11 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 12 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 13 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 14 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 15 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for adjusting PCSK9 comprises that using length is the short antisense compounds of 16 monomeric targeting PCSK9 nucleic acid.

In certain embodiments, the method for the expression of adjusting PCSK9 comprises that use comprises the short antisense compounds of 9-15 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for the expression of adjusting PCSK9 comprises that use comprises the short antisense compounds of 10-15 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for the expression of adjusting PCSK9 comprises that use comprises the short antisense compounds of 12-14 monomeric targeting PCSK9 nucleic acid.In certain embodiments, the method for the expression of adjusting PCSK9 comprises the short antisense compounds that uses the targeting PCSK9 nucleic acid that comprises 12-14 nucleotide or nucleoside.

4. superoxide dismutase 1 (SOD1)

Superoxide dismutase (SOD) can be by the catalysis superoxides to hydrogen peroxide (H₂O₂) disproportionation provide opposing to the oxidative damage of biomolecule (Fridovich, Annu.Rev.Biochem., 1995,64,97-112).Superoxide dismutase mainly contains two classes.One class is made up of one group of enzyme of the avtive spot with cupric and zinc, and another kind of avtive spot have manganese or ferrum (Fridovich, Annu.Rev.Biochem., 1995,64,97-112).

The sudden change in superoxide dismutase 1 gene and the amyotrophic lateral sclerosis of dominant inheritance's form (ALS also claims Lou Gehrig disease) are relevant, and the selectivity degeneration of the above lower motor neuron of this disease is feature (Cleveland and Liu, Nat.Med., 2000,6,1320-1321).Various sudden changes are likely that to the illeffects of superoxide dismutase 1 toxicity function rather than loss superoxide dismutase 1 activity mediate by promoting, can not cause obvious disease (Al-Chalabi and Leigh because neither can shorten the life-span when in mice, not having superoxide dismutase 1 fully yet, Curr.Opin.Neurol., 2000,13,397-405; Aliskyand Davidson, Hum.Gene Ther., 2000,11,2315-2329).

Identified 100 various mutations of people SOD1 gene, these sudden changes cause about 20% familial amyotrophic lateral sclerosis (ALS) case altogether.Some sudden changes (for example modal A4V sudden change of the U.S.) are highly lethal, only can survive 9 months from the disease symptoms outbreak.Other sudden changes of SOD1 show with slower disease process.

Definition

" SOD1 " means its expression and treats by lacking gene outcome or the protein that antisense compounds is regulated.

" SOD1 nucleic acid " means the nucleic acid of any coding SOD1.For example, in certain embodiments, SOD1 nucleic acid include but not limited to the to encode DNA sequence of SOD1, the RNA sequence of transcribing from the DNA of coding SOD1 and the mRNA sequence of coding SOD1.

" SOD1 mRNA " means the mRNA of coding SOD1.

SOD1 treats indication

Found that the Antisense Suppression to superoxide dismutase 1 (SOD1) can reduce SOD1 mRNA and protein simultaneously in the animal model of familial ALS, and further caused slowing down of progression of disease, the more important thing is that the time-to-live increases.Therefore, in certain embodiments, the invention provides by the short antisense compounds of targeting SOD1 nucleic acid being suffered from the individuality of familial ALS, the method for the progression of disease in the described individuality that slows down.In some this embodiment, the short antisense compounds of targeting SOD1 directly is delivered to the cerebrospinal fluid of individuality.In some this embodiment, described method also comprises the time-to-live that increases the individuality of suffering from familial ALS.Slowing down of progression of disease represented by the improvement of one or more indications of ALS progression of disease, and described indication includes but not limited to that modified ALS functional scale, pulmonary function test and muscle strength measure.

The SOD1 combination treatment

In certain embodiments, the pharmaceutical composition that one or more is comprised the short antisense compounds of targeting SOD1 nucleic acid gives altogether with one or more other medicament.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease identical with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease different with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament is designed to treat the ill effect of one or more pharmaceutical compositions of the present invention.In certain embodiments, one or more pharmaceutical compositions of the present invention are given altogether with another medicament, to treat the ill effect of this another medicament.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given simultaneously.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given at different time.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is prepared together in single dosage form.In certain embodiments, with other separately preparation of medicament of one or more pharmaceutical compositions of the present invention and one or more.

In certain embodiments, the medicament that gives altogether is a nicotinic acid.In some this embodiment, nicotinic acid is selected from rapid release nicotinic acid, extended release nicotinic acid and slow release nicotinic acid.

In certain embodiments, the medicament that gives altogether is a Carboxymethylcellulose.In some this embodiment, Carboxymethylcellulose is selected from gemfibrozil, fenofibrate, chlorine Bei Te, bezafibrate and ciprofibrate.

More examples of the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting SOD1 include but not limited to corticosteroid (including but not limited to prednisone); Immunoglobulin (includes but not limited to vein immunoglobulin (IVIg); Analgesic (for example acetaminophen); Antiinflammatory (including but not limited to NSAID (non-steroidal anti-inflammatory drug) (for example ibuprofen, COX-1 inhibitor and cox 2 inhibitor)); Salicylate; Antibiotic; Antiviral agent; Antifungal; Antidiabetic medicine (for example biguanide, glucosidase inhibitor, insulin, sulfonylureas and thiazolidinediones); The adrenergic modulation agent; Diuretic; Hormone (for example short anabolism steroid, androgen, estrogen, calcitonin, progesterone, somatostatin and thyroxin); Immunomodulator; Muscle relaxant; Hydryllin; Osteoporosis medicament (for example diphosphonate, calcitonin and estrogen); Prostaglandin; Antineoplastic agent; The spiritual healing medicament; Tranquilizer; Poison Oak Tree (oak) or black poison wood (sumac) product; Antibody and vaccine.

The short antisense compounds of some targeting SOD1 nucleic acid

In certain embodiments, lacking the antisense compounds targeting hasThe SOD1 nucleic acid of the sequence of searching number NM_X02317.1 (being incorporated herein) with SEQ ID NO:5.In some this embodiment, short antisense compounds and the SEQID NO:5 of targeting SEQ ID NO:5 have at least 90% complementarity.In some this embodiment, the short antisense compounds of targeting SEQ IDNO:5 and SEQ ID NO:5 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:5 of targeting SEQ ID NO:5 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:5 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 8 and 9.

The nucleotide sequence that each SEQ ID NO provides in the table 8 and 9 is independent of any to connecting the modification of key or nuclear base between sugar moieties, nucleoside.Thus, the defined short antisense compounds of certain SEQ ID NO can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 8 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:5.Table 8 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:5.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid can have short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3 and 1-9-2.

Table 8: the short antisense compounds of targeting SEQ ID NO:5

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						387541	85	100	GTCGCCCTTCAGCACG	3-10-3 MOE	406
387540	86	99	TCGCCCTTCAGCAC	2-10-2 MOE	407
						387539	87	98	CGCCCTTCAGCA	1-10-1 MOE	408

In certain embodiments, the target zone is the nucleotide 85-100 of SEQ ID NO:5.In some this embodiment, the short antisense compounds of the nucleotide 85-100 of targeting SEQ ID NO:5 comprises and is selected from SEQ ID NO:406,407 or 408 nucleotide sequence.In some this embodiment, the short antisense compounds of the nucleotide 85-100 of targeting SEQ ID NO:5 is selected from Isis No.387541,387540 or 387539.

In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting SOD1 nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting SOD1 nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting SOD1 nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating SOD1.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting SOD1 nucleic acid, and wherein the short antisense compounds of this targeting SOD1 nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting SOD1 nucleic acid is regulated the expression of SOD1.

In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 8 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 9 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 10 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 11 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 12 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 13 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 14 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 15 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for adjusting SOD1 comprises that using length is the short antisense compounds of 16 monomeric targeting SOD1 nucleic acid.

In certain embodiments, the method for the expression of adjusting SOD1 comprises that use comprises the short antisense compounds of 9-15 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for the expression of adjusting SOD1 comprises that use comprises the short antisense compounds of 10-15 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for the expression of adjusting SOD1 comprises that use comprises the short antisense compounds of 12-14 monomeric targeting SOD1 nucleic acid.In certain embodiments, the method for the expression of adjusting SOD1 comprises the short antisense compounds that uses the targeting SOD1 nucleic acid that comprises 12-14 nucleotide or nucleoside.

5.CRP

CRP (also claiming c-proteins C reactive and PTX1) is the multiple inflammatory cytokine of response and essential people's acute phase reactant of producing in liver.This protein of identifying as far back as nineteen thirty is high conservative, it is believed that it is the early stage indication of infectiousness or inflammatory disease.Plasma C RP level can respond infection, ischemia, wound, burn and inflammatory disease and increase by 1,000 times.In the patient accepts clinical trial that lipid reduces therapy (as the inhibin therapy), prove, the patient that LDL-C and CRP all reduce with only have LDL-C to reduce the patient to compare the risk mitigation of future development coronary event.

Definition

" CRP " means its expression and treats by lacking gene outcome or the protein that antisense compounds is regulated.

" CRP nucleic acid " means the nucleic acid of any coding CRP.For example, in certain embodiments, CRP nucleic acid include but not limited to the to encode DNA sequence of CRP, the RNA sequence of transcribing from the DNA of coding CRP and the mRNA of coding CRP.

" CRP mRNA " means the mRNA of coding CRP.

CRP treats indication

In certain embodiments, the invention provides the method for the CRP expression of regulating in the individuality, described method comprises that the short antisense compounds with targeting CRP nucleic acid gives this individuality.In certain embodiments, the invention provides the individual method of treatment, described method comprises and gives the pharmaceutical composition that one or more comprise the short antisense compounds of targeting CRP nucleic acid.In certain embodiments, this individuality suffers from hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, the familial hypercholesterolemia that isozygotys, Combination dyslipidemia, atherosclerosis, the atherosclerotic risk of development, coronary heart disease, coronary disease medical history, early show effect coronary heart disease, one or more coronary heart disease risk factors.In certain embodiments, this individuality suffers from acute coronary syndrome, blood vessel injury, arterial occlusion, unstable angina pectoris, back (post) peripheral blood vessel, back myocardial infarction (MI), thrombosis, deep venous thrombosis, end-stage renal disease (ESRD), chronic renal failure, complement activation, congestive heart failure or systemic vasculitis.In certain embodiments, this individuality suffers from apoplexy.

In certain embodiments, this individuality has carried out being selected from the operation of the implantation of optional (elective) support, angioplasty, back Percutaneous Transluminal Angioplasty (PTCA), heart transplantation, kidney dialysis or cardiopulmonary bypass.

In certain embodiments, this individuality suffers from inflammatory diseases.In some this embodiment, inflammatory diseases is selected from inflammatory bowel, ulcerative colitis, rheumatoid arthritis or osteoarthritis.

The guilding principle that lipid reduces therapy by u.s. national cholesterol education program (NCEP) adult treatment expert scheme III (ATPIII) in calendar year 2001 set up and upgraded in 2004 (Grundyet al., Circulation, 2004,110,227-239).This guilding principle comprises acquisition complete lipoprotein general picture (profile) after fasting in 9-12 hour usually, to determine LDL-C, T-CHOL and HDL-C level.According to the guilding principle of up-to-date formulation, 130-159mg/dL, 160-189mg/dL and think critical height, height and high level respectively more than or equal to the LDL-C level of 190mg/dL.200-239mg/dL and think critical height and high level respectively more than or equal to the total cholesterol level of 240mg/dL.HDL-C level less than 40mg/dL is thought low-level.

In certain embodiments, this individuality has been confirmed as carrying out lipid reduction therapy.In some this embodiment, according to upgrading (Grundyet al. in calendar year 2001 foundation and in 2004 by u.s. national cholesterol education program (NCEP) adult treatment expert scheme III (ATPIII), Circulation, 2004,110, guilding principle 227-239), this individuality have been confirmed as carrying out lipid and have reduced therapy.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 190mg/dL.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 160mg/dL.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 130mg/dL.In certain embodiments, need carry out its LDL-C of this individuality that lipid reduces therapy and be higher than 100mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 160mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 130mg/dL.In some this embodiment, this individuality that need carry out lipid reduction therapy should keep LDL-C to be lower than 100mg/dL.In some this embodiment, this individuality should keep LDL-C to be lower than 70mg/dL.

In certain embodiments, the invention provides the method that reduces the CRP in the individuality.In some this embodiment, CRP is reduced at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95% and at least 100%.

In certain embodiments, method provided by the invention does not reduce HDL-C.In certain embodiments, method provided by the invention does not cause the accumulation of lipid in liver.In certain embodiments, method provided by the invention does not cause fatty degeneration of liver.

In certain embodiments, can reduce again and the method for the treatment of relevant side effect when the invention provides CRP concentration in reducing the experimenter.In some this embodiment, side effect is a liver toxicity.In some this embodiment, side effect is an abnormal liver function.In some this embodiment, side effect is that alanine aminotransferase (ALT) raises.In some this embodiment, side effect is that aspartate aminotransferase (AST) raises.

In certain embodiments, the invention provides the method that reduces the CRP concentration among the experimenter not have to reach target LDL-C level because of lipid reduction therapy.In some this embodiment, the short antisense compounds of targeting CRP nucleic acid is that the unique lipid that gives the experimenter reduces medicament.In some this embodiment, experimenter's lipid that acceptance (comply with) recommended of not following the doctor's advice as yet reduces therapy.In some this embodiment, pharmaceutical composition of the present invention reduces therapy with in addition different lipid and gives altogether together.In some this embodiment, it is the LDL-Apheresis that other lipid reduces therapy.In some this embodiment, it is inhibin that other lipid reduces therapy.In some this embodiment, it is according to Ezetimibe that other lipid reduces therapy.

In certain embodiments, the invention provides the method for the CRP concentration among the experimenter that reduction do not tolerate inhibin.In some this embodiment, because given inhibin, experimenter's creatine kinase concentration improves.In some this embodiment, because given inhibin, experimenter's abnormal liver function.In some this embodiment, because given inhibin, experimenter's myalgia.In some this embodiment, because given inhibin, the side effect of experimenter's center of origin nervous system.In certain embodiments, experimenter's inhibin of accepting to be recommended of not following the doctor's advice as yet gives.

In certain embodiments, the invention provides the method that reduces the coronary heart disease risk among the experimenter.In certain embodiments, the invention provides the method for the atherosclerotic progress among the experimenter that slows down.In some this embodiment, the invention provides the method that stops the atherosclerotic progress among the experimenter.In some this embodiment, the invention provides the size of the atherosclerotic plaque that reduces among the experimenter and/or the method for diffusion.In certain embodiments, the method that is provided can reduce the experimenter and develop atherosclerotic risk.

In certain embodiments, the method that is provided can be improved the cardiovascular consequence among the experimenter.In some this embodiment, the improvement of cardiovascular consequence is that the risk of development coronary heart disease reduces.In some this embodiment, the improvement of cardiovascular consequence is the reduction of the incidence rate of one or more main cardiovascular events, and described incident includes but not limited to death, myocardial infarction, infraction, apoplexy, cardiogenic shock, pulmonary edema, cardiac arrest and atrial arrhythmia again.In some this embodiment, the improvement of cardiovascular consequence is confirmed by the improvement of carotid intimal medial thickness.In some this embodiment, the improvement of carotid intimal medial thickness is the reduction of thickness.In some this embodiment, the improvement of carotid intimal medial thickness is preventing of media thickness increase.

In certain embodiments, the pharmaceutical composition that comprises the short antisense compounds of targeting CRP nucleic acid is for using in treatment.In certain embodiments, described treatment is the reduction of the CRP in the individuality.In certain embodiments, described treatment is to following treatment of diseases: hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, the familial hypercholesterolemia that isozygotys, Combination dyslipidemia, atherosclerosis, the atherosclerotic risk of development, coronary heart disease, coronary disease medical history or the coronary heart disease of early showing effect.In certain embodiments, described treatment is the reduction of CRP risk.In certain embodiments, described treatment is atherosclerotic prevention.In certain embodiments, described treatment is the prevention of coronary heart disease.In certain embodiments, described treatment is to following treatment of diseases: acute coronary syndrome, chronic renal failure, blood vessel injury, arterial occlusion, atherosclerosis thrombosis, unstable angina pectoris, back (post) peripheral blood vessel, back myocardial infarction (MI), thrombosis, deep venous thrombosis, end-stage renal disease (ESRD), complement activation, congestive heart failure or systemic vasculitis.In certain embodiments, described treatment is to having carried out being selected from the implantation of optional (elective) support, angioplasty, the treatment of the individuality of the operation of Percutaneous Transluminal Angioplasty (PTCA), heart transplantation, kidney dialysis or cardiopulmonary bypass afterwards.In certain embodiments, described treatment is the treatment to inflammatory diseases.

In certain embodiments, the pharmaceutical composition that will comprise the short antisense compounds of targeting CRP nucleic acid is used for preparing the medicine in order to the CRP that reduces individuality.In certain embodiments, the pharmaceutical composition that will comprise the short antisense compounds of targeting CRP nucleic acid is used to prepare in order to reduce the medicine of coronary heart disease risk.In certain embodiments, the short antisense compounds with targeting CRP nucleic acid is used to prepare in order to treat the medicine of following disease: hypercholesterolemia, non-familial hypercholesterolemia, familial hypercholesterolemia, heterozygous familial hypercholesterolemia, the familial hypercholesterolemia that isozygotys, Combination dyslipidemia, atherosclerosis, the atherosclerotic risk of development, coronary heart disease, coronary disease medical history, early show effect coronary heart disease or one or more coronary heart disease risk factors.

In certain embodiments, the short antisense compounds with targeting CRP nucleic acid is used to prepare in order to treat the medicine of following disease: acute coronary syndrome, chronic renal failure, blood vessel injury, arterial occlusion, atherosclerosis thrombosis, unstable angina pectoris, back (post) peripheral blood vessel, back myocardial infarction (MI), thrombosis, deep venous thrombosis, end-stage renal disease (ESRD), complement activation, congestive heart failure or systemic vasculitis.In certain embodiments, the short antisense compounds with targeting CRP nucleic acid is used to prepare the medicine of suffering from the individuality of apoplexy in order to treatment.

In certain embodiments, the short antisense compounds of targeting CRP nucleic acid is used for preparation has carried out being selected from the operation of the implantation of optional (elective) support, angioplasty, back Percutaneous Transluminal Angioplasty (PTCA), heart transplantation, kidney dialysis or cardiopulmonary bypass in order to treatment the medicine of individuality.

In certain embodiments, the short antisense compounds with targeting CRP nucleic acid is used to prepare in order to treat the medicine of inflammatory diseases.In certain embodiments, the short antisense compounds with targeting CRP nucleic acid is used to prepare in order to treat the medicine of inflammatory bowel, ulcerative colitis, rheumatoid arthritis or osteoarthritis.

The CRP combination treatment

In certain embodiments, the pharmaceutical composition that one or more is comprised the short antisense compounds of targeting CRP nucleic acid gives altogether with one or more other medicament.In certain embodiments, described one or more other medicament is that lipid reduces medicament.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease identical with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease different with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament is designed to treat the ill effect of one or more pharmaceutical compositions of the present invention.In certain embodiments, one or more pharmaceutical compositions of the present invention are given altogether with another medicament, to treat the ill effect of this another medicament.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given simultaneously.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given at different time.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is prepared together in single dosage form.In certain embodiments, with other separately preparation of medicament of one or more pharmaceutical compositions of the present invention and one or more.

In certain embodiments, the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting CRP nucleic acid comprises that lipid reduces medicament.In some this embodiment, can include but not limited to atorvastatin, simvastatin, rosuvastatin with the medicament that pharmaceutical composition of the present invention gives altogether and according to Ezetimibe.In some this embodiment, lipid is reduced medicament give before the pharmaceutical composition of the present invention giving.In some this embodiment, lipid is reduced medicament after giving pharmaceutical composition of the present invention, give.In some this embodiment, lipid is reduced medicament when giving pharmaceutical composition of the present invention, give.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is identical with the dosage that gives separately to be given when lipid reduces medicament.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is lower than the dosage that is given when giving lipid reduction medicament separately.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is higher than the dosage that is given when giving lipid reduction medicament separately.

In certain embodiments, it is the HMG-CoA reductase inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, the HMG-CoA reductase inhibitor is an inhibin.In some this embodiment, inhibin is selected from atorvastatin, simvastatin, pravastatin, fluvastatin and rosuvastatin.

In certain embodiments, it is ISIS 301012 that the lipid that gives altogether reduces medicament.

In certain embodiments, it is cholesterol absorption inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, cholesterol absorption inhibitor is according to Ezetimibe.

In certain embodiments, it is HMG-CoA reductase inhibitor and cholesterol absorption inhibitor formulated together altogether that the lipid that gives altogether reduces medicament.In some this embodiment, it is according to Ezetimibe/simvastatin that lipid formulated together altogether reduces medicament.

In certain embodiments, it is microsomal triglyceride transfer protein inhibitor (MTP inhibitor) that the lipid that gives altogether reduces medicament.

In certain embodiments, the medicament that gives altogether is a bile acid chelating agent.In some this embodiment, bile acid chelating agent is selected from colestyramine, colestipol and colesevelam.

In certain embodiments, the medicament that gives altogether is a nicotinic acid.In some this embodiment, nicotinic acid is selected from rapid release nicotinic acid, extended release nicotinic acid and slow release nicotinic acid.

In certain embodiments, the medicament that gives altogether is a Carboxymethylcellulose.In some this embodiment, Carboxymethylcellulose is selected from gemfibrozil, fenofibrate, chlorine Bei Te, bezafibrate and ciprofibrate.

More examples of the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting CRP nucleic acid include but not limited to corticosteroid (including but not limited to prednisone); Immunoglobulin (includes but not limited to vein immunoglobulin (IVIg); Analgesic (for example acetaminophen); Antiinflammatory (including but not limited to NSAID (non-steroidal anti-inflammatory drug) (for example ibuprofen, COX-1 inhibitor and cox 2 inhibitor)); Salicylate; Antibiotic; Antiviral agent; Antifungal; Antidiabetic medicine (for example biguanide, glucosidase inhibitor, insulin, sulfonylureas and thiazolidinediones); The adrenergic modulation agent; Diuretic; Hormone (for example short anabolism steroid, androgen, estrogen, calcitonin, progesterone, somatostatin and thyroxin); Immunomodulator; Muscle relaxant; Hydryllin; Osteoporosis medicament (for example diphosphonate, calcitonin and estrogen); Prostaglandin; Antineoplastic agent; The spiritual healing medicament; Tranquilizer; Poison Oak Tree (oak) or black poison wood (sumac) product; Antibody and vaccine.

In certain embodiments, the pharmaceutical composition that comprises the short antisense compounds of targeting CRP nucleic acid can be reduced therapy in conjunction with lipid gives.In some this embodiment, it is that curative life style changes that lipid reduces therapy.In some this embodiment, it is the LDL Apheresis that lipid reduces therapy.

The short antisense compounds of some targeting CRP nucleic acid

In certain embodiments, lacking the antisense compounds targeting hasThe CRP nucleic acid of the sequence of searching number NM_000567.1 (being incorporated herein) with SEQ ID NO:6.In some this embodiment, short antisense compounds and the SEQ IDNO:6 of targeting SEQ ID NO:6 have at least 90% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:6 of targeting SEQ ID NO:6 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:6 of targeting SEQ ID NO:6 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:6 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in the table 9.

The nucleotide sequence that each SEQ ID NO provides in the table 9 is independent of any to connecting the modification of key or nuclear base between sugar moieties, nucleoside.Thus, the defined short antisense compounds of certain SEQ ID NO can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 9 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:6.Table 9 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:6.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

In certain embodiments, the short antisense compounds of targeting CRP nucleic acid can have short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting CRP nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3 and 1-9-2.

Table 9: the short antisense compounds of targeting SEQ ID NO:6

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	Seq ID NO
						353506	1257	1272	ACTCTGGACCCAAACC	3-10-3 MOE	409
353507	1258	1271	CTCTGGACCCAAAC	2-10-2 MOE	410
						353484	1305	1320	CCATTTCAGGAGACCT	3-10-3 MOE	411
353485	1306	1319	CATTTCAGGAGACC	2-10-2 MOE	412

In certain embodiments, the target zone is the nucleotide 1305-1320 of NM_000567.1.In some this embodiment, the short antisense compounds of the nucleotide 1305-1320 of targeting NM_000567.1 comprises the nucleotide sequence that is selected from SEQ ID NO:1305 or 1306.In some this embodiment, the short antisense compounds of the nucleotide 263-278 of targeting NM_000567.1 is selected from Isis NO.353484 or 353485.

In certain embodiments, the target zone is the nucleotide 1257-1272 of NM_000567.1.In some this embodiment, the short antisense compounds of the nucleotide 1257-1272 of targeting NM_000567.1 comprises the nucleotide sequence that is selected from SEQ ID NO 1257 or 1258.In some this embodiment, the short antisense compounds of the nucleotide 428-483 of targeting NM_000567.1 is selected from Isis NO.353506 or 353507.

In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting CRP nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting CRP nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting CRP nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting CRP nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting CRP nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting CRP nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting CRP nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting CRP nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting CRP nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting CRP nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting CRP nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating CRP.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting CRP nucleic acid, and wherein the short antisense compounds of this targeting CRP nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting CRP nucleic acid is regulated the expression of CRP.

In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 8 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 9 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 10 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 11 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 12 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 13 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 14 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 15 monomeric targeting CRP nucleic acid.In certain embodiments, the method for adjusting CRP comprises that using length is the short antisense compounds of 16 monomeric targeting CRP nucleic acid.

In certain embodiments, the method for the expression of adjusting CRP comprises that use comprises the short antisense compounds of 9-15 monomeric targeting CRP nucleic acid.In certain embodiments, the method for the expression of adjusting CRP comprises that use comprises the short antisense compounds of 10-15 monomeric targeting CRP nucleic acid.In certain embodiments, the method for the expression of adjusting CRP comprises that use comprises the short antisense compounds of 12-14 monomeric targeting CRP nucleic acid.In certain embodiments, the method for the expression of adjusting CRP comprises the short antisense compounds that uses the targeting CRP nucleic acid that comprises 12-14 nucleotide or nucleoside.

6. glycocorticosteroid receptor (GCCR)

Glucocorticoid is to obtain one of steroid hormone of identifying the earliest, be responsible for different physiological roles, including but not limited to the synthetic inhibition of inhibition, cytokine of the sedimentary increase of reduction, glycogen, immunity and the inflammatory reaction of glucose uptake and utilization in the stimulation, surrounding tissue of glyconeogenesis and the acceleration of various growth incidents.Glucocorticoid for resist stress particular importance also.The rising synthetic and that discharge of the glucocorticoid of stress-induced can cause various replying, comprise the increase that increase, the inhibition of inflammatory mediator, the synthetic inhibition of cytokine and the glucose of ventricle workload produce (Karin, Cell, 1998,93,487-490).

Natural glucocorticoid and their synthesis of derivatives all are to apply its effect by glycocorticosteroid receptor, and this receptor is the omnipresence express cell matter member of the nuclear hormone superfamily of receptor.People's glycocorticosteroid receptor also claims nuclear receptor superfamily 3, group C, and the member 1; NR3C1; GCCR; GCR; GRL; Glycocorticosteroid receptor, lymphocyte.Its gene is positioned on the human chromosome 5q11-q13, forms (Encio andDetera-Wadleigh, J Biol Chem, 1991,266,7182-7188 by 9 exons; Gehring et al., Proc Natl Acad Sci USA, 1985,82,3751-3755).Exist people's glycocorticosteroid receptor mRNA of various ways: the 5.5kb people's glycocorticosteroid receptor α cDNA that contains exons 1-8 and exon 9 α; The 4.3kb people's glycocorticosteroid receptor β cDNA that contains exons 1-8 and exon 9 β; With the 7.0kb people's glycocorticosteroid receptor α cDNA that contains exons 1-8 and whole exon 9, it comprises exon 9 α, exon 9 β and " J district ", and " J district " flank is exon 9 α and 9 β (Hollenberg et al., Nature, 1985,318,635-641; Oakley et al., J Biol Chem, 1996,271,9550-9559).People's glycocorticosteroid receptor α is the main isotype of receptor, be show steroid in conjunction with active isotype (Hollenberg et al., Nature, 1985,318,635-641).In addition, by using three different promoteres, can transcribe out three different exons 1 variants, the alternately montage of an exons 1 variant can cause producing three different editions of this exon.Therefore, people's glycocorticosteroid receptor mRNA can contain 5 different editions exons 1 (Breslin et al., MolEndocrinol, 2001,15,1381-1395).

Research to the expression pattern of the α of people's glycocorticosteroid receptor mRNA and β isotype discloses, and the expression of α isotype is horn of plenty more.Two kinds of isotypes comprise lung, kidney, heart, liver, skeletal muscle, macrophage, neutrophil cell and peripheral blood lymphocytes all similarly expressing in tissue and the cell type.Only there is people's glycocorticosteroid receptor α in colon, to express.On protein level, be subjected to all to detect the α isotype in the inspection tissue at all, and the β isotype can not detect, this prompting is under physiological condition, the montage approach of acquiescence is approach α isotype (Pujols et al., Am J Physiol Cell Physiol, 2002 that produce, 283, C1324-1331).The β isotype of glycocorticosteroid receptor is neither in conjunction with the glucocorticoid agonist, also not in conjunction with the glucocorticoid antagonist.In addition, the β isotype mainly is positioned in the nucleus of transfected cell, does not rely on hormonal stimulation.When two kinds of isotypes are all expressed in same cell, the gene expression that inhibitory hormone is inductive, glycocorticosteroid receptor is alpha mediated of glycocorticosteroid receptor β energy stimulates, this prompting β isotype plays effect (the Oakley et al. of the inhibitor of glycocorticosteroid receptor alpha active, J Biol Chem, 1996,271,9550-9559).Unless otherwise, otherwise people's glycocorticosteroid receptor described herein is defined as the omnipresence product of the gene that is positioned on the chromosome 5q11-q13.

Transfection the cell line of complementary glycocorticosteroid receptor antisense RNA chain, demonstrate that glycocorticosteroid receptor mRNA level reduces and glycocorticosteroid receptor agonist dexamethasone replied reduction (Pepin and Barden, Mol Cell Biol, 1991,11,1647-1653).The transgenic mice that carries antisense glycocorticosteroid receptor gene constructs be used to study glucocorticoid to the feedback effect of hypothalmus-pituitary-adrenal axis (Pepin etal., Nature, 1992,355,725-728).In another research to similar genetically engineered mice, caloric intake and expenditure, heart and vastus lateralis lipoprotein lipase activity and heart and brown adipose tissue norepinephrine are all compared according to mice low.On the contrary, fat content and total body energy comparison are significantly much higher according to mice.These results suggest, defective glycocorticosteroid receptor west is carried can influence the energy balance by increasing energy efficiency, and these results have given prominence to regulating action (the Richard et al. of hypothalmus-pituitary-adrenal axis variation to the muscle lipoprotein lipase activity, Am J Physiol, 1993,265, R146-150).

Assess in the animal model of anxiety, learning and memory power being designed, measured the influence to behavior of glycocorticosteroid receptor antagonist.The minimizing that glycocorticosteroid receptor is expressed in the rat of the antisense oligodeoxyribonucleotide of long-term Intraventricular infusion targeting glycocorticosteroid receptor mRNA, do not disturb space in the Morris water maze laboratory (the Engelmann et al. that cruises, Eur J Pharmacol, 1998,361,17-26).The antisense oligodeoxyribonucleotide of bilateral infusion targeting glycocorticosteroid receptor mRNA in the dentate gyrus of rat hippocampus, stationarity (immobility) (Korte et al., Eur J Pharmacol, 1996 of rat in the experiment of Porsolt forced swimming have been reduced, 301,19-25).

Glucocorticoid usually is because it uses obtaining such as the immunosuppressant in the treatment of diseases of anaphylaxis, asthma, rheumatoid arthritis, AIDS, systemic lupus erythematosus (sle) and degenerative osteoarthritis, antiinflammatory action.The negative adjusting of gene expression as the negative adjusting that the interaction by glycocorticosteroid receptor and NF-kB causes, it is believed that be the reason of glucocorticoid antiinflammatory action in vivo to small part.These three kinds of cytokines of interleukin-6, tumor necrosis factor and interleukin-1 are the reasons that cause most of hypothalamic-pituitary-adrenals (HPA) axle of inflammation in stress process to stimulate.Hpa axis and systemic sympathetic nerve and adrenal medulla system are the peripheral components of stress response system, be responsible for keeping basic stable state with stress relevant stable state.Glucocorticoid is as the end products of hpa axis, can suppress the generation of all three kinds of inflammatory cytokine, can also suppress their effects to target tissue, exception be interleukin-6, itself and glucocorticoid synergism stimulate the generation of acute phase reactant.The activity of glucocorticoid treatment can reduction hpa axis (Chrousos, N Engl J Med, 1995,332,1351-1362).

In some cases, the patient can resist the glucocorticoid treatment.A reason of the resistance of this a pair of steroid is the sudden change or the pleomorphism that exist in the glycocorticosteroid receptor gene.Be reported in and had 15 missense, 3 nonsenses, 3 frameshit, 1 splice site and 2 road montages sudden change (alternative splice mutation) and 16 relevant (Bray and Cotton of pleomorphism in addition in the NR3C1 gene with the glucocorticoid resistance, Hum Mutat, 2003,21,557-568).The other research of in the mankind, carrying out pointed out positive correlation between the allele of metabolism syndrome incidence rate and progress and glycocorticosteroid receptor (GR) gene (Rosmond, Obes Res, 2002,10,1078-1086).

The glucocorticoid insensitivity of other situations is relevant with the change of the expression of glycocorticosteroid receptor isotype.The research that people's glycocorticosteroid receptor β isotype mRNA among the glucocorticoid resistance ulcerative colitis patient is expressed discloses, the existence of this mRNA is than significantly higher in the responsive patient of glucocorticoid, the expression of this prompter's glycocorticosteroid receptor β mRNA in peripheral blood lymphocytes can be served as predictor (the Honda et al. that the glucocorticoid in the ulcerative colitis is replied, Gastroenterology, 2000,118,859-866).Increase in obviously numerous expression of glucocorticoid also being observed glycocorticosteroid receptor β in insensitive asthmatic patient.In addition, in peripheral blood lymphocytes from insensitive patient's transfection to glucocorticoid, found DNA binding ability unusual of the glycocorticosteroid receptor that causes by cytokine, and the HepG2 cell with glycocorticosteroid receptor β gene has caused remarkable reduction (the Leung et al. of glycocorticosteroid receptor α DNA binding ability, J Exp Med, 1997,186,1567-1574).The dexamethasone combination studies have shown that people's glycocorticosteroid receptor β does not change the affinity of glycocorticosteroid receptor α to the hormone part, but changes its ability in conjunction with GRE (Bamberger et al., J ClinInvest, 1995,95,2435-2441).Generally speaking, these presentation of results, glycocorticosteroid receptor β by with glycocorticosteroid receptor α competition GRE target site, can serve as on the physiology with Pathophysiology on relevant endogenous inhibitor to the glucocorticoid effect.

In liver, the glucocorticoid agonist increases the liver glucose generation by activating glycocorticosteroid receptor, and this can cause the expression of glyconeogenesis enzyme PCK (PEPCK) and G-6-Pase to increase subsequently.By glyconeogenesis, glucose be able to from non-hexose precursor such as lactic acid, acetone acid and alanine form (Link, Curr Opin InvestigDrugs, 2003,4,421-429).Steroid glycocorticosteroid receptor antagonist such as RU 486 in the rodent model of diabetes, have been tested.The mice (being called the db/db mice) that lacks the leptin acceptor gene is obesity in heredity, suffer from diabetes and hyperinsulinemia.It is about 49% to have reduced blood-glucose with the db/db mice of RU 486 treatment hyperglycemia, does not influence plasma insulin level.In addition, compare with untreated mice, expression (the Friedman et al. of glycocorticosteroid receptor response gene PEPCK, G-6-Pase, glucose transporter 2 types and the tyrosine aminotransferase of RU 486 treatments reduction the in the db/db mice, J Biol Chem, 1997,272,31475-31481).RU 486 can also be by reducing diabetes (the Gettys et al. in the ob/ob mouse model that serum insulin and blood glucose levels improve diabetes, obesity and hyperinsulinemia, Int J Obes Relat Metab Disord, 1997,21,865-873).

Because the increase of glyconeogenesis is considered to the main source of the increase that glucose produces in the diabetes, done research to multiple for the treatment target that suppresses the liver glucose generation.Because the antagonist of glycocorticosteroid receptor can improve diabetes in animal model, this chemical compound is one of potential medicine of being sought.But, the glycocorticosteroid receptor antagonist has deleterious systemic effect, comprise hpa axis activation (Link, Curr Opin InvestigDrugs, 2003,4,421-429).The active increase of hpa axis is relevant with the inhibition of immune related inflammation effect, and this suppresses to increase the susceptibility to infectious agent and tumor.The relevant disease of inhibition with the immunomediated inflammatory that causes by the defective in hpa axis or its target tissue, comprise Cushing syndrome, chronic stress, chronic alcoholism and depressive type melancholy (Chrousos, NEngl J Med, 1995,332,1351-1362).Therefore, be opened to liver specificity glycocorticosteroid receptor antagonist and can have value especially.Steroid glycocorticosteroid receptor antagonist and bile acid are puted together, so that with their targeting livers (Apelqvist et al., 2000).The at present known healing potion that does not also have energy targeting glycocorticosteroid receptor and don't unwanted periphery influence occurs (Link, Curr Opin Investig Drugs, 2003,4,421-429).Therefore, to the demand of the medicament that can effectively suppress liver glycocorticosteroid receptor long-term existence still.

Definition

" glycocorticosteroid receptor " is that its expression is treated by lacking gene outcome or the protein that antisense compounds is regulated.Glycocorticosteroid receptor is commonly referred to GCCR.

" GCCR nucleic acid " means the nucleic acid of any coding GCCR.For example, in certain embodiments, GCCR nucleic acid include but not limited to the to encode DNA sequence of GCCR, the RNA sequence of transcribing from the DNA of coding GCCR and the mRNA sequence of coding GCCR." GCCRmRNA " means the mRNA of coding GCCR.

The treatment indication

Antisense technology is the effective means that reduces the expression of specific gene product, therefore can be used for multiple in order in therapeutic, diagnostic and the research application of regulating the glycocorticosteroid receptor expression.In addition, in certain embodiments, liver be give antisense oligonucleotide find to exist one of tissue of maximum concentration (Geary et al., Curr.Opin.Investig.Drugs, 2001,2,562-573).Therefore, the method that in this embodiment, that antisense technology has been represented is noticeable, the liver specificity of glycocorticosteroid receptor is suppressed.

In certain embodiments, the short antisense compounds of the nucleic acid of targeting coding glycocorticosteroid receptor is arrived liver by precedence partition.In certain embodiments, lacking the usefulness that antisense compounds compares with longer parent compound in liver improves.In certain embodiments, target RNA is mainly expressed in liver.

Use for treatment, suffer from the disease that to treat by the expression of regulating GCCR or the experimenter of disease by giving one or more short antisense compounds, treat to suspect.In limiting examples, described method comprises that the short antisense compounds with the treatment effective dose gives the step of animal.Some short antisense compounds can suppress the active of GCCR and/or suppress the expression of GCCR.In certain embodiments, the active or expression of the GCCR among the experimenter is suppressed and reaches at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100%.In certain embodiments, the active or expression of the GCCR among the experimenter is suppressed and reaches at least 30%.In certain embodiments, the active or expression of the GCCR among the experimenter is suppressed and reaches at least 50% or more.

The reduction of the expression of GCCR can for example be measured in blood, blood plasma, serum, fatty tissue, liver or any other body fluid, tissue or the organ of this animal.In certain embodiments, contained cell includes the nucleic acid of the GCCR that encodes and/or they contain GCCR albumen itself in the middle of this fluid to be analyzed, tissue or the organ.

Some medicine that comprises short antisense compounds and other compositionss also are provided.In certain embodiments, short antisense compounds is to be added to suitable medicine by certain chemical compound with effective dose can accept diluent or carrier, uses in pharmaceutical composition.

In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid has short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-10-1,2-10-2,3-10-3 and 1-9-2.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 2-10-2 and 2-8-2.

In certain embodiments, this paper provides by short antisense compounds that gives one or more targeting GCCR nucleic acid or the pharmaceutical composition that comprises this chemical compound and treats individual method.Also provide by the short antisense compounds that gives targeting GCCR nucleic acid and treat the method for suffering from the experimenter of active diseases associated of GCCR or disease.Except diabetes particularly the type ii diabetes, include but not limited to obesity with GCCR diseases associated and disease, metabolic syndrome X, Cushing syndrome, the Ai Dixunshi disease, inflammatory diseases such as asthma, rhinitis and arthritis, anaphylaxis, autoimmune disease, immunodeficiency, anorexia, cachexia, bone loss or bone fragility and wound healing.Metabolic syndrome, metabolic syndrome X or be called for short syndrome X, be meant comprise that obesity, dyslipidemia (particularly high blood triglyceride), glucose do not tolerate, one group of risk factor of hyperglycemia and hypertension.In certain embodiments, the short antisense compounds with targeting GCCR is used for improving the hyperglycemia that is caused by systemic Steroid treatment.In addition, antisense technology provides the means of the expression that suppresses glycocorticosteroid receptor β isotype, and this isotype is proved to be overexpression in the patient who resists the glucocorticoid treatment.

In certain embodiments, the invention provides targeting coding GCCR nucleic acid, can regulate the short antisense compounds of the expression of glycocorticosteroid receptor.Medicine and other compositionss of comprising The compounds of this invention also are provided.The method of regulator of screening glycocorticosteroid receptor and the method for regulating the expression of glycocorticosteroid receptor in cell, tissue or animal are provided in addition, and described method comprises makes described cell, tissue or animal contact with one or multiple The compounds of this invention or compositions.This paper gives treatment and suspects to suffer from or easily suffer from method with the animal (particularly people) of the expression diseases associated or the disease of glycocorticosteroid receptor.This method comprises treatment or prevents one or more The compounds of this invention of effective dose or the people that compositions needs treatment.

The short antisense compounds of some targeting GCCR nucleic acid

In certain embodiments, lacking the antisense compounds targeting has

The GCCR nucleic acid of the sequence of the nucleotide 1-106000 of searching number AC012634 (being incorporated herein) with SEQ ID NO:8.In some this embodiment, short antisense compounds and the SEQ ID NO:8 of targeting SEQ ID NO:8 have at least 90% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:8 of targeting SEQ ID NO:8 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:8 of targeting SEQ ID NO:8 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:8 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 10 and 11.

The nucleotide sequence that each SEQ ID NO provides in the table 10 and 11 is independent of any to connecting the modification of key or nuclear base between sugar moieties, nucleoside.Thus, the defined short antisense compounds of certain SEQ ID NO can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises the gapmer die body.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises 2-10-2 gapmer die body.

Table 10 and 11 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:8.Table 10 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:8.Table 11 has been enumerated the short antisense compounds that has one or two mispairing with respect to SEQ ID NO:8.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Table 10: the short antisense compounds of targeting SEQ ID NO:8

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						371644	88142	88155	TTTGGGAGGTGGTC	2-10-2 MOE	413
371645	88156	88169	CACACCAGGCAGAG	2-10-2 MOE	414
						371649	88212	88225	CTTTACAGCTTCCA	2-10-2 MOE	415
371651	88242	88255	CACTACCTTCCACT	2-10-2 MOE	416
						371652	88248	88261	AACACACACTACCT	2-10-2 MOE	417
371653	88256	88269	CTCTTCAAAACACA	2-10-2 MOE	418
						371665	92037	92050	GTAATTGTGCTGTC	2-10-2 MOE	419
371669	92086	92099	TTTTTCTTCGAATT	2-10-2 MOE	420
						371671	92114	92127	CATTTTCGATAGCG	2-10-2 MOE	421
371673	92142	92155	ACCTTCCAGGTTCA	2-10-2 MOE	422

Table 11: targeting SEQ ID NO:8 and have the short antisense compounds of 1 or 2 mispairing

ISIS NO	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						371638	2039	2052	ATAGGAAGCATAAA	2-10-2 MOE	423
371650	4949	4962	TCTTTTAAAGAAGA	2-10-2 MOE	424
						371673	10187	10200	ACCTTCCAGGTTCA	2-10-2 MOE	422
371660	13465	13478	AAGGATATTTTAAA	2-10-2 MOE	425
						371660	14428	14441	AAGGATATTTTAAA	2-10-2 MOE	425
371654	15486	15499	GAACAAAAATTAAA	2-10-2 MOE	427
						371661	16638	16651	TTCCACAGATCTGT	2-10-2 MOE	428
371653	17892	17905	CTCTTCAAAACACA	2-10-2 MOE	418
						371679	18444	18457	TTTATAAAGTAAAG	2-10-2 MOE	429
371645	19816	19829	CACACCAGGCAGAG	2-10-2 MOE	414
						371638	21555	21568	ATAGGAAGCATAAA	2-10-2 MOE	423
371650	21775	21788	TCTTTTAAAGAAGA	2-10-2 MOE	424
						371679	21902	21915	TTTATAAAGTAAAG	2-10-2 MOE	429
371655	22507	22520	TACTGTGAGAAATA	2-10-2 MOE	433
						371655	22722	22735	TACTGTGAGAAATA	2-10-2 MOE	433
371672	25662	25675	TTCCAGCTTGAAGA	2-10-2 MOE	435
						371678	25926	25939	GATCAGTTCTCATG	2-10-2 MOE	436
371655	26041	26054	TACTGTGAGAAATA	2-10-2 MOE	433
						371638	29770	29783	ATAGGAAGCATAAA	2-10-2 MOE	423

371668	30551	30564	TTATCAATGATGCA	2-10-2 MOE	439
						371670	40584	40597	GCATGCTGGACAGT	2-10-2 MOE	440
371654	43331	43344	GAACAAAAATTAAA	2-10-2 MOE	427
						371650	46024	46037	TCTTTTAAAGAAGA	2-10-2 MOE	424
371659	50372	50385	TTGCACCTGAACTA	2-10-2 MOE	443
						371634	50565	50578	CAGAATATATTTCT	2-10-2 MOE	444
371673	56942	56955	ACCTTCCAGGTTCA	2-10-2 MOE	422
						371654	62372	62385	GAACAAAAATTAAA	2-10-2 MOE	427
371679	63537	63550	TTTATAAAGTAAAG	2-10-2 MOE	429
						371654	64908	64921	GAACAAAAATTAAA	2-10-2 MOE	427
371661	65795	65808	TTCCACAGATCTGT	2-10-2 MOE	428
						371645	70997	71010	CACACCAGGCAGAG	2-10-2 MOE	414
371661	77400	77413	TTCCACAGATCTGT	2-10-2 MOE	428
						371663	82329	82342	ATAAGAGATTAAAA	2-10-2 MOE	450
371633	83426	83439	TCCCCCTTCTCATT	2-10-2 MOE	451
						371662	85873	85886	GGGCATTGTTAAAA	2-10-2 MOE	452
371654	86476	86489	GAACAAAAATTAAA	2-10-2 MOE	427
						371679	86516	86529	TTTATAAAGTAAAG	2-10-2 MOE	429
371641	88097	88110	AGAACTCACATCTG	2-10-2 MOE	455
						371642	88111	88124	GAGCTGGACGGAGG	2-10-2 MOE	456
371646	88170	88183	AAGCTTCATCGGAG	2-10-2 MOE	457
						371647	88184	88197	ATAATGGCATCCCG	2-10-2 MOE	458
371650	88226	88239	TCTTTTAAAGAAGA	2-10-2 MOE	424
						371673	91493	91506	ACCTTCCAGGTTCA	2-10-2 MOE	422
371664	92030	92043	TGCTGTCCTATAAG	2-10-2 MOE	460
						371666	92044	92057	CACAAAGGTAATTG	2-10-2 MOE	461
371667	92058	92071	ATCATTTCTTCCAG	2-10-2 MOE	462
						371668	92072	92085	TTATCAATGATGCA	2-10-2 MOE	463
371670	92100	92113	GCATGCTGGACAGT	2-10-2 MOE	440
						371672	92128	92141	TTCCAGCTTGAAGA	2-10-2 MOE	435
371674	92147	92160	CCATTACCTTCCAG	2-10-2 MOE	466
						371637	92983	92996	GCATAAACAGGGTT	2-10-2 MOE	467
371654	93928	93941	GAACAAAAATTAAA	2-10-2 MOE	427
						371641	99772	99785	AGAACTCACATCTG	2-10-2 MOE	455
371679	99883	99896	TTTATAAAGTAAAG	2-10-2 MOE	429
						371660	99933	99946	AAGGATATTTTAAA	2-10-2 MOE	425
371635	105004	105017	TATGAAAGGAATGT	2-10-2 MOE	472
						371654	105028	105041	GAACAAAAATTAAA	2-10-2 MOE	427
371676	106482	106495	TTCCTTAAGCTTCC	2-10-2 MOE	474
						371650	107838	107851	TCTTTTAAAGAAGA	2-10-2 MOE	424
371673	110922	110935	ACCTTCCAGGTTCA	2-10-2 MOE	422
						371673	111580	111593	ACCTTCCAGGTTCA	2-10-2 MOE	422
371634	114608	114621	CAGAATATATTTCT	2-10-2 MOE	444
						371638	115040	115053	ATAGGAAGCATAAA	2-10-2 MOE	423
371660	116244	116257	AAGGATATTTTAAA	2-10-2 MOE	425
						371663	116657	116670	ATAAGAGATTAAAA	2-10-2 MOE	450
371673	118068	118081	ACCTTCCAGGTTCA	2-10-2 MOE	422
						371666	118834	118847	CACAAAGGTAATTG	2-10-2 MOE	461
371660	119858	119871	AAGGATATTTTAAA	2-10-2 MOE	425
						371660	120210	120223	AAGGATATTTTAAA	2-10-2 MOE	425
371662	120876	120889	GGGCATTGTTAAAA	2-10-2 MOE	452

371655	124004	124017	TACTGTGAGAAATA	2-10-2 MOE	433
						371656	124170	124183	GAACAGTTAAACAT	2-10-2 MOE	485

In certain embodiments, the target zone is the nucleotide 88142-88269 of SEQ ID NO:8.In certain embodiments, lack the nucleotide 88142-88269 of antisense compounds targeting SEQ ID NO:8.In some this embodiment, the short antisense compounds of targeted nucleotide 88142-88269 comprises the nucleotide sequence that is selected from SEQ ID NO 413,414,415,416,417 or 418.In some this embodiment, the short antisense compounds of the nucleotide 88142-88269 of targeting SEQ IDNO:8 is selected from Isis NO.371644,371645,371649,371651,371652 or 371653.

In certain embodiments, the target zone is the nucleotide 88142-88169 of SEQ ID NO:8.In certain embodiments, lack the nucleotide 88142-88169 of antisense compounds targeting SEQ ID NO:8.In some this embodiment, the short antisense compounds of targeted nucleotide 88142-88169 comprises the nucleotide sequence that is selected from SEQ ID NO 413 or 414.In some this embodiment, the short antisense compounds of the nucleotide 88142-88169 of targeting SEQ ID NO:8 is selected from Isis NO.371644 or 371645.

In certain embodiments, the target zone is the nucleotide 88242-88269 of SEQ ID NO:8.In certain embodiments, lack the nucleotide 88242-88269 of antisense compounds targeting SEQ ID NO:8.In some this embodiment, the short antisense compounds of targeted nucleotide 88242-88269 comprises the nucleotide sequence that is selected from SEQ ID NO 416,417 or 418.In some this embodiment, the short antisense compounds of the nucleotide 88242-88269 of targeting SEQ ID NO:8 is selected from Isis NO.371651,371652 or 371653.

In certain embodiments, the target zone is the nucleotide 92037-92155 of SEQ ID NO:8.In certain embodiments, lack the nucleotide 92037-92155 of antisense compounds targeting SEQ ID NO:8.In some this embodiment, the short antisense compounds of targeted nucleotide 92037-92155 comprises the nucleotide sequence that is selected from SEQ ID NO 419,420,421 or 422.In some this embodiment, the short antisense compounds of the nucleotide 92037-92155 of targeting SEQ ID NO:8 is selected from Isis NO.371665,371669,371671 or 171673.

In certain embodiments, the target zone is the nucleotide 92114-92155 of SEQ ID NO:8.In certain embodiments, lack the nucleotide 92114-92155 of antisense compounds targeting SEQ ID NO:8.In some this embodiment, the short antisense compounds of targeted nucleotide 92114-92155 comprises the nucleotide sequence that is selected from SEQ ID NO 421 or 422.In some this embodiment, the short antisense compounds of the nucleotide 92114-92155 of targeting SEQ ID NO:8 is selected from Isis NO.371671 or 171673.

In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting GCCR nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting GCCR nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting GCCR nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating GCCR.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting GCCR nucleic acid, and wherein the short antisense compounds of this targeting GCCR nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting GCCR nucleic acid is regulated the expression of GCCR.

In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 8 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 9 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 10 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 11 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for adjusting GCCR comprises the short antisense compounds that uses 12 monomeric targeting GCCR nucleic acid of length.In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 13 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 14 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 15 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for adjusting GCCR comprises that using length is the short antisense compounds of 16 monomeric targeting GCCR nucleic acid.

In certain embodiments, the method for the expression of adjusting GCCR comprises that use comprises the short antisense compounds of 9-15 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for the expression of adjusting GCCR comprises that use comprises the short antisense compounds of 10-15 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for the expression of adjusting GCCR comprises that use comprises the short antisense compounds of 12-14 monomeric targeting GCCR nucleic acid.In certain embodiments, the method for the expression of adjusting GCCR comprises the short antisense compounds that uses the targeting GCCR nucleic acid that comprises 12-14 nucleotide or nucleoside.

7. glucagon receptor (GCGR)

Normal keeping of glucemia is the metabolism incident of a careful adjusting.This is responsible for keeping in postabsorptive state (postabsorbative state) 29 amino acid peptides of blood glucose levels glucagon, can increase the release of glucose by activating the lipolysis in liver glycogenolysis, glyconeogenesis, the stimulation fatty tissue and stimulating insulin secretion from liver.In hypertension glucose level process, insulin can reverse the liver glycogenolysis of glucagon mediation and the enhancing of glyconeogenesis.In diabetics, insulin or do not have, or not exclusively effectively.Though treatment of diabetes is concentrating on the increase insulin level traditionally, but the antagonism of glucagon function has been considered to a kind of alternative medicine because glucagon is to bring into play its physiological action by carrying out signal transduction with the nationality glucagon receptor, therefore glucagon receptor has been considered to potential treatment target (the Madsen et al. of diabetes, Curr.Pharm.Des., 1999,5,683-691).

Glucagon receptor belongs to the superfamily of the g protein coupled receptor with seven membrane spaning domains.It still can in conjunction with structurally with the member of the littler subfamily of the homoreceptor of the similar peptide of glucagon.The gene of coding human glucagon receptor was cloned in 1994, to the analysis of genome sequence disclosed a plurality of introns and with 82% homogeneity (Lok et al., Gene, 1994,140, the 203-209. of rat glucagon receptor gene; MacNeil et al., Biochem.Biophys.Res.Commun., 1994,198,328-334).The clone of rat glucagon receptor gene also caused multiple in addition road splice variant report (Maget et al., FEBS Lett., 1994,351,271-275).Human glucagon receptor gene mapping on chromosome 17q25 (Menzel et al., Genomics, 1994,20,327-328).Codon 40 Gly of place cause the affinity of glucagon is reduced by 3 times of (Fujisawa et al. to the missense mutation of Ser in glucagon receptor gene, Diabetologia, 1995,38,983-985), and this sudden change is associated with several morbid states, comprise noninsulindependent diabetes (Fujisawa et al., Diabetologia, 1995,38,983-985), hypertension (Chambers andMorris, Nat.Genet., 1996,12,122) and central obesity (centraladiposity) (Siani et al., Obes.Res., 2001,9,722-726).

Definition

" glucagon receptor " is that its expression is treated by lacking gene outcome or the protein that antisense compounds is regulated.Glucagon receptor is commonly referred to GCGR, but also can be described as GR, GGR, MGC138246, MGC93090.

" GCGR nucleic acid " means the nucleic acid of any coding GCGR.For example, in certain embodiments, GCGR nucleic acid include but not limited to encode GCGR (DNA) sequence of GCGR, the RNA sequence of transcribing from the DNA of coding GCGR and the mRNA sequence of coding GCGR." GCGR mRNA " means the proteic mRNA of coding GCGR.

The treatment indication

Antisense technology is to reduce the effective means that glucagon receptor (GCGR) is expressed, and has been proved to be in multiple therapeutic, diagnosis type and the application of research property to have unique applications.Thus, in certain embodiments, the invention provides targeting coding glucagon receptor nucleic acid, can regulate the short antisense compounds of the expression of glucagon receptor.This paper provides in addition and can suppress the short antisense compounds that GCGR expresses.This paper also provides treatment individual method, and described method comprises and gives the pharmaceutical composition that one or more comprise the short antisense compounds of targeting GCGR nucleic acid.In certain embodiments, because suppressing GCGR, the short antisense compounds of targeting GCGR nucleic acid expresses, this paper provides by giving the pharmaceutical composition that one or more comprise the short antisense compounds of targeting GCGR nucleic acid, treat to suffer from the method with the experimenter of active diseases associated of GCGR or disease.For example, this paper provides treatment to suffer from the experimenter's of high blood-glucose, hyperglycemia, prediabetes, type ii diabetes, metabolic syndrome, obesity and/or insulin resistance method.

This paper also is susceptible to the pharmaceutical composition of short antisense compounds that comprises one or more targeting GCGR and drug acceptable carrier, diluent, reinforcing agent or the excipient of choosing wantonly.Some The compounds of this invention also can be used for preparing in order to treatment and the glucagon effect diseases associated of GCGR mediation and the medicine of disease.

Certain embodiments of the present invention comprise the method for the expression that reduces the GCGR in tissue or the cell, and described method comprises contacts the encode short antisense compounds of nucleic acid of GCGR or the pharmaceutical composition that comprises this short antisense compounds of described cell or tissue and targeting.In some this embodiment, the invention provides the method that reduces blood glucose levels, blood triglyceride level or blood cholesterol levels among the experimenter, described method comprises and gives this experimenter with weak point antisense compounds or pharmaceutical composition.Blood levels can be blood plasma level or serum levels.What also be susceptible to is method, the method that improves the GLP-1 level of improving insulin sensitivity in the animal and the method that suppresses liver glucose output, and these methods comprise and give described animal with antisense oligonucleotide of the present invention or pharmaceutical composition.The improvement of insulin sensitivity can be represented by the reduction of circulation insulin level.

In certain embodiments, the invention provides treatment and suffer from method with the experimenter of the GLA diseases associated of GCGR mediation or disease, described method comprise will treatment or the short antisense compounds or the pharmaceutical composition of prevention effective dose give this experimenter.In certain embodiments, this disease or disease may be metabolic disease or disease.In certain embodiments, this metabolic disease or disease are diabetes, hyperglycemia, hyperlipemia, metabolic syndrome X, obesity, the high glucagonemia of constitutional, insufficient insulin or insulin resistance.In some embodiments, diabetes are type ii diabetes.In some embodiments, obesity is that meals cause.In some embodiments, hyperlipemia raises relevant with the blood lipid level.Lipid comprises cholesterol and triglyceride.In one embodiment, disease is the liver fat degeneration.In some embodiments, steatosis is fat hepatitis or non-alcoholic stellato-hepatitis.

In certain embodiments, the invention provides and use oligomeric compounds described herein, prevent or postpone the method and the method that keeps the β cell function in the animal of the outbreak that blood glucose levels raises in the animal.

The short antisense compounds of some targeting GCGR can be used to regulate the expression of the GCGR among the experimenter's (as animal, including but not limited to the people) who needs.In certain embodiments, this method comprises that the short antisense compounds with the expression that can reduce GCGR RNA of effective dose gives the step of described animal.In certain embodiments, lack level or the function that antisense compounds can reduce GCGRRNA effectively.Because the reduction of GCGR mRNA level also can cause the change of GCGR protein expression product, also can be measured this change that produces.Some can effectively reduce the level of GCGR RNA or protein expression product or the antisense compounds of function, is considered to active antisense compounds.In certain embodiments, short antisense compounds can reduce the expression of GCGR, causes RNA to reduce and reaches at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100%.

The method of regulator of screening glucagon receptor and the method for regulating the expression of glucagon receptor in cell, tissue or animal are provided in addition, described method comprise the short antisense compounds that makes described cell, tissue or animal and one or multiple targeting GCGR or with comprise this compound compositions and contact.This paper gives treatment and suspects to suffer from or easily suffer from method with the animal (particularly people) of the expression diseases associated or the disease of glucagon receptor.Some this method comprises treatment or prevents one or more The compounds of this invention of effective dose or the people that compositions needs treatment.

The reduction of the expression of glucagon receptor can for example be measured in blood, blood plasma, serum, fatty tissue, liver or any other body fluid, tissue or the organ of this animal.Preferably, contained cell contains coding proteic nucleic acid of glucagon receptor and/or glucagon receptor albumen itself in the middle of this analyzed fluid, tissue or the organ.

Medicine and other compositionss of comprising short antisense compounds also are provided.In certain embodiments, the short antisense compounds of nucleic acid of targeting coding GCGR is to be added to suitable medicine by certain chemical compound with effective dose can accept diluent or carrier, uses in pharmaceutical composition.

The short antisense compounds of targeting GCGR nucleic acid has short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,2-10-2,3-10-3,3-8-3,1-1-10-2.

The short antisense compounds of some targeting GCGR nucleic acid

In certain embodiments, lack the antisense compounds targeting and have sequence

The GCGR nucleic acid of searching number NM_000160.1 (being incorporated herein) with SEQ ID NO:9.In some this embodiment, short antisense compounds and the SEQ IDNO:9 of targeting SEQ ID NO:9 have at least 90% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:9 of targeting SEQ ID NO:9 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:9 of targeting SEQ ID NO:9 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:9 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 12 and 13.

The nucleotide sequence that each SEQ ID NO provides in the table 12 and 13 is independent of any to connecting the modification of key or nuclear base between sugar moieties, nucleoside.Thus, the defined short antisense compounds of certain SEQ ID NO can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises the gapmer die body.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 3-10-3 gapmer die body.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises the gapmer die body.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 3-8-3 gapmer die body.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises the gapmer die body.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 2-10-2 gapmer die body.

Table 12 and 13 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:9.Table 12 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:9.Table 13 has been enumerated the short antisense compounds that has one or two mispairing with respect to SEQ ID NO:9.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Table 12: the short antisense compounds of targeting SEQ ID NO:9

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						338463	378	393	TAGAGCTTCCACTTCT	3-10-3 MOE	486
338534	378	391	GAGCTTCCACTTCT	3-8-3 MOE	487
						327130	499	512	TGTTGGCCGTGGTA	3-8-3 MOE	488
327131	500	513	ATGTTGGCCGTGGT	3-8-3 MOE	489
						327132	501	514	GATGTTGGCCGTGG	3-8-3 MOE	490
327133	502	515	AGATGTTGGCCGTG	3-8-3 MOE	491
						327134	503	516	GAGATGTTGGCCGT	3-8-3 MOE	492
327135	504	517	GGAGATGTTGGCCG	3-8-3 MOE	493
						327136	505	518	AGGAGATGTTGGCC	3-8-3 MOE	494
327137	506	519	CAGGAGATGTTGGC	3-8-3 MOE	495
						327138	507	520	GCAGGAGATGTTGG	3-8-3 MOE	496
327139	508	521	GGCAGGAGATGTTG	3-8-3 MOE	497
						327140	531	544	GTGGTGCCAAGGCA	3-8-3 MOE	498
327141	532	545	TGTGGTGCCAAGGC	3-8-3 MOE	499
						327142	533	546	TTGTGGTGCCAAGG	3-8-3 MOE	500
327143	534	547	TTTGTGGTGCCAAG	3-8-3 MOE	501
						327144	535	548	CTTTGTGGTGCCAA	3-8-3 MOE	502
327145	536	549	ACTTTGTGGTGCCA	3-8-3 MOE	503
						327146	537	550	CACTTTGTGGTGCC	3-8-3 MOE	504
327147	538	551	GCACTTTGTGGTGC	3-8-3 MOE	505
						327148	539	552	TGCACTTTGTGGTG	3-8-3 MOE	506

327149	540	553	TTGCACTTTGTGGT	3-8-3 MOE	507
						327150	545	558	CGGTGTTGCACTTT	3-8-3 MOE	508
327151	546	559	GCGGTGTTGCACTT	3-8-3 MOE	509
						327152	547	560	AGCGGTGTTGCACT	3-8-3 MOE	510
327153	548	561	AAGCGGTGTTGCAC	3-8-3 MOE	511
						327154	549	562	GAAGCGGTGTTGCA	3-8-3 MOE	512
327155	550	563	CGAAGCGGTGTTGC	3-8-3 MOE	513
						327156	551	564	ACGAAGCGGTGTTG	3-8-3 MOE	514
327157	552	565	CACGAAGCGGTGTT	3-8-3 MOE	515
						327158	553	566	ACACGAAGCGGTGT	3-8-3 MOE	516
327159	554	567	AACACGAAGCGGTG	3-8-3 MOE	517
						345897	684	697	GCTGCTGTACATCT	2-10-2 MOE	518
327160	684	697	GCTGCTGTACATCT	3-8-3 MOE	518
						327161	685	698	AGCTGCTGTACATC	3-8-3 MOE	520
327162	686	699	AAGCTGCTGTACAT	3-8-3 MOE	521
						327163	687	700	GAAGCTGCTGTACA	3-8-3 MOE	522
327164	688	701	GGAAGCTGCTGTAC	3-8-3 MOE	523
						327165	689	702	TGGAAGCTGCTGTA	3-8-3 MOE	524
327166	690	703	CTGGAAGCTGCTGT	3-8-3 MOE	525
						327167	691	704	CCTGGAAGCTGCTG	3-8-3 MOE	526
327168	692	705	ACCTGGAAGCTGCT	3-8-3 MOE	527
						327169	693	706	CACCTGGAAGCTGC	3-8-3 MOE	528
327170	694	707	TCACCTGGAAGCTG	3-8-3 MOE	529
						327171	695	708	ATCACCTGGAAGCT	3-8-3 MOE	530
327172	696	709	CATCACCTGGAAGC	3-8-3 MOE	531
						327173	697	710	ACATCACCTGGAAG	3-8-3 MOE	532
327174	698	711	TACATCACCTGGAA	3-8-3 MOE	533
						327175	699	712	GTACATCACCTGGA	3-8-3 MOE	534
327176	700	713	TGTACATCACCTGG	3-8-3 MOE	535
						327177	701	714	GTGTACATCACCTG	3-8-3 MOE	536
327178	869	882	TAGCGGGTCCTGAG	3-8-3 MOE	537
						327179	870	883	GTAGCGGGTCCTGA	3-8-3 MOE	538
327180	871	884	TGTAGCGGGTCCTG	3-8-3 MOE	539
						327181	872	885	CTGTAGCGGGTCCT	3-8-3 MOE	540
327182	873	886	GCTGTAGCGGGTCC	3-8-3 MOE	541
						327183	874	887	GGCTGTAGCGGGTC	3-8-3 MOE	542
327184	875	888	TGGCTGTAGCGGGT	3-8-3 MOE	543
						327185	876	889	CTGGCTGTAGCGGG	3-8-3 MOE	544
327186	877	890	TCTGGCTGTAGCGG	3-8-3 MOE	545
						327187	878	891	TTCTGGCTGTAGCG	3-8-3 MOE	546
327188	955	968	TGAACACCGCGGCC	3-8-3 MOE	547
						327189	956	969	ATGAACACCGCGGC	3-8-3 MOE	548
327190	957	970	CATGAACACCGCGG	3-8-3 MOE	549
						327191	958	971	GCATGAACACCGCG	3-8-3 MOE	550
327192	959	972	TGCATGAACACCGC	3-8-3 MOE	551
						327193	960	973	TTGCATGAACACCG	3-8-3 MOE	552
327194	961	974	ATTGCATGAACACC	3-8-3 MOE	553
						327195	962	975	TATTGCATGAACAC	3-8-3 MOE	554
327196	963	976	ATATTGCATGAACA	3-8-3 MOE	555
						327197	964	977	CATATTGCATGAAC	3-8-3 MOE	556
327198	1019	1032	AGGTTGTGCAGGTA	3-8-3 MOE	557

327199	1020	1033	CAGGTTGTGCAGGT	3-8-3 MOE	558
						327200	1021	1034	GCAGGTTGTGCAGG	3-8-3 MOE	559
327201	1022	1035	AGCAGGTTGTGCAG	3-8-3 MOE	560
						327202	1023	1036	CAGCAGGTTGTGCA	3-8-3 MOE	561
327203	1024	1037	CCAGCAGGTTGTGC	3-8-3 MOE	562
						327204	1025	1038	CCCAGCAGGTTGTG	3-8-3 MOE	563
327205	1026	1039	GCCCAGCAGGTTGT	3-8-3 MOE	564
						327206	1027	1040	GGCCCAGCAGGTTG	3-8-3 MOE	565
327207	1028	1041	AGGCCCAGCAGGTT	3-8-3 MOE	566
						338491	1160	1175	TGTCATTGCTGGTCCA	3-10-3 MOE	567
338562	1160	1173	TCATTGCTGGTCCA	3-8-3 MOE	568
						338498	1307	1322	TGGCCAGCCGGAACTT	3-10-3 MOE	569
338569	1307	1320	GCCAGCCGGAACTT	3-8-3 MOE	570
						338499	1329	1344	GGGATGAGGGTCAGCG	3-10-3 MOE	571
338570	1329	1342	GATGAGGGTCAGCG	3-8-3 MOE	572
						385067	1364	1377	AAGGCAAAGACCAC	3-8-3 MOE	573
338573	1401	1414	GGAGCGCAGGGTGC	3-8-3 MOE	574
						338580	1487	1500	TGCACCTCCTTGTT	3-8-3 MOE	575

Table 13: targeting SEQ ID NO:9 and have the short antisense compounds of 1 or 2 mispairing

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						338577	158	171	CAGCAGACCCTGGA	3-8-3 MOE	576
338458	237	252	ACATCTGGCAGAGGTT	3-10-3 MOE	577
						338529	237	250	ATCTGGCAGAGGTT	3-8-3 MOE	578
338466	318	333	CAGGCCAGCAGGAGTA	3-10-3 MOE	579
						338537	318	331	GGCCAGCAGGAGTA	3-8-3 MOE	580
338533	364	377	CAAACAAAAAGTCC	3-8-3 MOE	582
						338462	364	379	CTCAAACAAAAAGTCC	3-10-3 MOE	581
338535	397	410	GGTGACATTGGTCA	3-8-3 MOE	584
						338464	397	412	GTGGTGACATTGGTCA	3-10-3 MOE	583
338466	470	485	CAGGCCAGCAGGAGTA	3-10-3 MOE	579
						338537	470	483	GGCCAGCAGGAGTA	3-8-3 MOE	580
385048	497	510	TTGGCAGTGGTGTT	3-8-3 MOE	587
						385049	500	513	ATGTTGGCAGTGGT	3-8-3 MOE	588
338467	503	518	AGGAAATGTTGGCAGT	3-10-3 MOE	589
						338538	503	516	GAAATGTTGGCAGT	3-8-3 MOE	590
385050	506	519	CAGGAAATGTTGGC	3-8-3 MOE	591
						385051	509	522	GGGCAGGAAATGTT	3-8-3 MOE	592
385052	523	536	AAGGTAGGTACCAG	3-8-3 MOE	593
						385053	526	539	ACCAAGGTAGGTAC	3-8-3 MOE	594
385056	535	548	CTTTGTGGCACCAA	3-8-3 MOE	595
						385057	538	551	GCACTTTGTGGCAC	3-8-3 MOE	596
338539	539	552	TGCACTTTGTGGCA	3-8-3 MOE	597
						385058	541	554	GCTGCACTTTGTGG	3-8-3 MOE	598
385059	544	557	GGTGCTGCACTTTG	3-8-3 MOE	599
						385060	547	560	GGCGGTGCTGCACT	3-8-3 MOE	600

385063	556	569	TGAACACTAGGCGG	3-8-3 MOE	601
						385064	559	572	TCTTGAACACTAGG	3-8-3 MOE	602
338469	561	576	CACCTCTTGAACACTA	3-10-3 MOE	603
						338540	561	574	CCTCTTGAACACTA	3-8-3 MOE	604
385065	562	575	ACCTCTTGAACACT	3-8-3 MOE	605
						385066	565	578	CACACCTCTTGAAC	3-8-3 MOE	606
338541	590	603	CCTCGAACCCACTG	3-8-3 MOE	607
						338473	658	673	CTTCTGGACCTCGATC	3-10-3 MOE	608
338544	658	671	TCTGGACCTCGATC	3-8-3 MOE	609
						338474	681	696	CTGCTATACATCTTGG	3-10-3 MOE	610
338545	681	694	GCTATACATCTTGG	3-8-3 MOE	611
						338475	703	718	CACGGTGTACATCACC	3-10-3 MOE	612
338546	703	716	CGGTGTACATCACC	3-8-3 MOE	613
						338547	718	731	ACAGACTGTAGCCC	3-8-3 MOE	615
338476	718	733	GGACAGACTGTAGCCC	3-10-3 MOE	614
						338550	889	902	CATCGCCAATCTTC	3-8-3 MOE	617
338479	889	904	GTCATCGCCAATCTTC	3-10-3 MOE	616
						338551	899	912	ACACTGAGGTCATC	3-8-3 MOE	619
338480	899	914	TCACACTGAGGTCATC	3-10-3 MOE	618
						338552	924	937	CGCCCCGTCACTGA	3-8-3 MOE	620
338555	992	1005	AGCAACCAGCAATA	3-8-3 MOE	622
						338484	992	1007	CCAGCAACCAGCAATA	3-10-3 MOE	621
338485	1018	1033	CAGGCTGTACAGGTAC	3-10-3 MOE	623
						338556	1018	1031	GGCTGTACAGGTAC	3-8-3 MOE	624
338558	1051	1064	AGCTCCTCTCAGAG	3-8-3 MOE	626
						338487	1051	1066	GAAGCTCCTCTCAGAG	3-10-3 MOE	625
338559	1079	1092	CAGCCAATGCCCAG	3-8-3 MOE	628
						338488	1079	1094	CCCAGCCAATGCCCAG	3-10-3 MOE	627
338560	1131	1144	AAACAGACACTTGA	3-8-3 MOE	630
						338489	1131	1146	TCAAACAGACACTTGA	3-10-3 MOE	629
338490	1145	1160	AGCACTGAACATTCTC	3-10-3 MOE	631
						338561	1145	1158	CACTGAACATTCTC	3-8-3 MOE	632
338563	1181	1194	ATCCACCAGAATCC	3-8-3 MOE	634
						338492	1181	1196	GGATCCACCAGAATCC	3-10-3 MOE	633
338564	1216	1229	TGATCAGTAAGGCC	3-8-3 MOE	635
						338565	1232	1245	ACAAAGATGAAAAA	3-8-3 MOE	637
338494	1232	1247	GGACAAAGATGAAAAA	3-10-3 MOE	636
						338566	1267	1280	CACGCAGCTTGGCC	3-8-3 MOE	639
338495	1267	1282	GGCACGCAGCTTGGCC	3-10-3 MOE	638
						338571	1344	1357	GACCCCCAGCAGAG	3-8-3 MOE	641
338500	1344	1359	TGGACCCCCAGCAGAG	3-10-3 MOE	640
						385068	1366	1379	CAAAGGCAAAGACC	3-8-3 MOE	642
385069	1369	1382	TCACAAAGGCAAAG	3-8-3 MOE	643
						385070	1372	1385	CAGTCACAAAGGCA	3-8-3 MOE	644
385071	1375	1388	CGTCAGTCACAAAG	3-8-3 MOE	645
						385072	1378	1391	GCTCGTCAGTCACA	3-8-3 MOE	646
385073	1381	1394	CATGCTCGTCAGTC	3-8-3 MOE	647
						386608	1384	1397	GGGCATGCTCGTCA	1-12-1 MOE	648

386593	1384	1397	GGGCATGCTCGTCA	2-10-2 MOE	648
						396146	1384	1397	GGGCATGCTCGTCA	2-10-2 MOE	648
338572	1384	1397	GGGCATGCTCGTCA	3-8-3 MOE	648
						396149	1384	1397	GGGCATGCTCGTCA	1-1-10-2 2 '-(butyl acetamido)-palmitamide/OMe/OMe	648
386627	1384	1397	GGGCATGCTCGTCA	2-10-2 methylene oxygen base BNA	648
						386610	1387	1400	CTTGGGCATGCTCG	1-12-1 MOE	654
386595	1387	1400	CTTGGGCATGCTCG	2-10-2 MOE	654
						385074	1387	1400	CTTGGGCATGCTCG	3-8-3 MOE	654
385075	1390	1403	TGCCTTGGGCATGC	3-8-3 MOE	657
						385076	1393	1406	GGGTGCCTTGGGCA	3-8-3 MOE	648
385077	1396	1409	GCAGGGTGCCTTGG	3-8-3 MOE	659
						385078	1399	1412	AGCGCAGGGTGCCT	3-8-3 MOE	660
338502	1401	1416	GTGGAGCGCAGGGTGC	3-10-3 MOE	661
						385079	1402	1415	TGGAGCGCAGGGTG	3-8-3 MOE	662
385080	1405	1418	TGGTGGAGCGCAGG	3-8-3 MOE	663
						385081	1408	1421	GCTTGGTGGAGCGC	3-8-3 MOE	664
385082	1411	1424	AGAGCTTGGTGGAG	3-8-3 MOE	665
						338503	1412	1427	AAAAGAGCTTGGTGGA	3-10-3 MOE	666
338574	1412	1425	AAGAGCTTGGTGGA	3-8-3 MOE	667
						385083	1414	1427	AAAAGAGCTTGGTG	3-8-3 MOE	668
385084	1417	1430	CAAAAAAGAGCTTG	3-8-3 MOE	669
						338504	1434	1449	AAGGAGCTGAGGAACA	3-10-3 MOE	670
338575	1434	1447	GGAGCTGAGGAACA	3-8-3 MOE	671
						327167	1441	1454	CCTGGAAGCTGCTG	3-8-3 MOE	526
338576	1445	1458	AGACCCTGGAAGGA	3-8-3 MOE	673
						338505	1445	1460	GCAGACCCTGGAAGGA	3-10-3 MOE	672
338506	1449	1464	ACCAGCAGACCCTGGA	3-10-3 MOE	674
						338577	1449	1462	CAGCAGACCCTGGA	3-8-3 MOE	576
338507	1464	1479	CAGTAGAGAACAGCCA	3-10-3 MOE	676
						338578	1464	1477	GTAGAGAACAGCCA	3-8-3 MOE	677
338508	1475	1490	TGTTGAGGAAACAGTA	3-10-3 MOE	678
						338579	1475	1488	TTGAGGAAACAGTA	3-8-3 MOE	679
338509	1487	1502	CCTGCACCTCCTTGTT	3-10-3 MOE	680
						338580	1610	1623	TGCACCTCCTTGTT	3-8-3 MOE	575

In certain embodiments, the target zone is the nucleotide 378-391 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 378-391 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 378-391 comprises the nucleotide sequence that is selected from SEQ ID NO 486 or 487.In some this embodiment, the short antisense compounds of the nucleotide 378-391 of targeting SEQ ID NO:9 is selected from IsisNo 338463 or 338534.

In certain embodiments, the target zone is the nucleotide 499-521 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 499-521 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 499-521 comprises the nucleotide sequence that is selected from SEQ ID NO 488,489,490,491,492,493,494,495,496 or 497.In some this embodiment, the short antisense compounds of the nucleotide 499-521 of targeting SEQ ID NO:9 is selected from Isis No 327130,327131,327132,327133,327134,327135,327136,327137,327138 or 327139.

In certain embodiments, the target zone is the nucleotide 531-553 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 531-553 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 531-553 comprises the nucleotide sequence that is selected from SEQ ID NO 498,499,500,501,502,503,504,505,506 or 507.In some this embodiment, the short antisense compounds of the nucleotide 531-553 of targeting SEQ ID NO:9 is selected from Isis No 327140,327141,327142,327143,327144,327145,327146,327147,327148 or 327149.

In certain embodiments, the target zone is the nucleotide 545-567 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 545-567 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 545-567 comprises the nucleotide sequence that is selected from SEQ ID NO 508,509,510,511,512,513,514,515,516 or 517.In some this embodiment, the short antisense compounds of the nucleotide 545-567 of targeting SEQ ID NO:9 is selected from Isis No 327150,327151,327152,327153,327154,327155,327156,327157,327158 or 327159.

In certain embodiments, the target zone is the nucleotide 531-567 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 531-567 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 531-567 comprises the nucleotide sequence that is selected from SEQ ID NO 498,499,500,501,502,503,504,505,506,507,508,509,510,511,512,513,514,515,516 or 517.In some this embodiment, the short antisense compounds of the nucleotide 531-567 of targeting SEQ ID NO:9 is selected from Isis No 327140,327141,327142,327143,327144,327145,327146,327147,327148,327149,327150,327151,327152,327153,327154,327155,327156,327157,327158 or 327159.

In certain embodiments, the target zone is the nucleotide 684-714 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 684-714 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 684-714 comprises the nucleotide sequence that is selected from SEQ ID NO 518,520,521,522,523,524,525,526,527,528,529,530,531,532,533,534,535 or 536.In some this embodiment, the short antisense compounds of the nucleotide 684-714 of targeting SEQ ID NO:9 is selected from Isis No 345897,327160,327161,327162,327163,327164,327165,327166,327167,327168,327169,327170,327171,327172,327173,327174,327175,327176 or 327177.

In certain embodiments, the target zone is the nucleotide 869-891 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 869-891 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 869-891 comprises the nucleotide sequence that is selected from SEQ ID NO 537,538,539,540,541,542,543,544,545 or 546.In some this embodiment, the short antisense compounds of the nucleotide 869-891 of targeting SEQ ID NO:9 is selected from Isis No 327178,327179,327180,327181,327182,327183,327184,327185,327186 or 327187.

In certain embodiments, the target zone is the nucleotide 955-977 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 955-977 of antisense compounds targeting SEQ ID NO:9.9. in some this embodiment, the short antisense compounds of targeted nucleotide 955-977 comprises the nucleotide sequence that is selected from SEQ ID NO 547,548,549,550,551,552,553,554,555 or 556.In some this embodiment, the short antisense compounds of the nucleotide 955-977 of targeting SEQ IDNO:9 is selected from Isis No 327188,327189,327190,327191,327192,327193,327194,327195,327196 or 327197.

In certain embodiments, the target zone is the nucleotide 1019-1041 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 1019-1041 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 1019-1041 comprises the nucleotide sequence that is selected from SEQ ID NO 557,558,559,560,561,562,563,564,565 or 566.In some this embodiment, the short antisense compounds of the nucleotide 1019-1041 of targeting SEQ ID NO:9 is selected from Isis No327198,327199,327200,327201,327202,327203,327204,327205,327206 or 327207.

In certain embodiments, the target zone is the nucleotide 1160-1175 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 1160-1175 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 1160-1175 comprises the nucleotide sequence that is selected from SEQ ID NO 567 or 568.In some this embodiment, the short antisense compounds of the nucleotide 1160-1175 of targeting SEQ ID NO:9 is selected from Isis No 338491 or 338562.

In certain embodiments, the target zone is the nucleotide 1307-1377 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 1307-1377 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 1307-1377 comprises the nucleotide sequence that is selected from SEQ ID NO 569,570,571,572 or 573.In some this embodiment, the short antisense compounds of the nucleotide 1307-1377 of targeting SEQ ID NO:9 is selected from Isis No 338498,338569,338499,338570 or 385067.

In certain embodiments, the target zone is the nucleotide 1307-1414 of SEQ ID NO:9.In certain embodiments, lack the nucleotide 1307-1414 of antisense compounds targeting SEQ ID NO:9.In some this embodiment, the short antisense compounds of targeted nucleotide 1307-1414 comprises the nucleotide sequence that is selected from SEQ ID NO 569,570,571,572,573 or 574.In some this embodiment, the short antisense compounds of the nucleotide 1307-1414 of targeting SEQ ID NO:9 is selected from Isis No 338498,338569,338499,338570,385067 or 338573.

In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting GCGR nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting GCGR nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting GCGR nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating GCGR.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting GCGR nucleic acid, and wherein the short antisense compounds of this targeting GCGR nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting GCGR nucleic acid is regulated the expression of GCGR.

In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 8 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 9 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 10 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 11 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 12 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 13 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 14 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 15 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for adjusting GCGR comprises that using length is the short antisense compounds of 16 monomeric targeting GCGR nucleic acid.

In certain embodiments, the method for the expression of adjusting GCGR comprises that use comprises the short antisense compounds of 9-15 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for the expression of adjusting GCGR comprises that use comprises the short antisense compounds of 10-15 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for the expression of adjusting GCGR comprises that use comprises the short antisense compounds of 12-14 monomeric targeting GCGR nucleic acid.In certain embodiments, the method for the expression of adjusting GCGR comprises the short antisense compounds that uses the targeting GCGR nucleic acid that comprises 12-14 nucleotide or nucleoside.

8.DGAT2

DG transferring enzyme 2 (also claims DGAT2, DG O-transferring enzyme 2, acetyl-CoA: DG Acetylase 2) be proved and relate to the absorption process of triglyceride (also claiming triacylglycerol) from food.

Triglyceride is a very efficient process from the absorption of food, and it is undertaken by series of steps, meals triacylglycerol hydrolysis in enteric cavity in these steps, then in the middle of enterocyte by resynthesis.The resynthesis of triacylglycerol can be undertaken by the monoacylglycerol approach, and this approach is by monoacylglycerol acyltransferase (MGAT) catalysis diacylglycerol synthetic from monoacylglycerol and fatty acyl group-CoA.The alternative synthetic method of DG is provided by the phosphoglycerol approach, and this approach relates to bimolecular fatty acyl group-CoA and glycerol-3-phosphate coupling.In arbitrary situation, acidylate all takes place with the fatty acyl group-CoA of another molecule in DG then in by the catalytic reaction of one of two kinds of diacylglycerol acyltransferases, forms triglyceride (Farese et al.; Curr.Opin.Lipidol.; 2000,11,229-234).

By the catalytic reaction of diacylglycerol acyltransferase is the last and unique critical step of triglyceride in synthetic.Thus, diacylglycerol acyltransferase relates to the fat storage in intestinal fat absorption, lipoprotein assembling, plasma triglyceride concentration adjusting and the adipose cell.First kind of diacylglycerol acyltransferase is that DG transferring enzyme 1 was identified in nineteen sixty, encode this proteinic people and little musculus cdna separated (Cases et al., Proc.Natl.Acad.Sci.U.S.A., 1998 in 1998,95,13018-13023; Oelkers et al., J.Biol.Chem., 1998,273,26765-26771).The mice that lacks diacylglycerol acyltransferase 1 can survive, and still can pass through other biological approach synthetic glycerine three esters, this prompting exist multiple triglyceride synthesis mechanism (Smith et al., Nat.Genet., 2000,25,87-90).

Second kind of DG transferring enzyme is that DG transferring enzyme 2 (also claims DGAT2, DG O-transferring enzyme 2, acyl group-CoA: diacylglycerol acyltransferase 2) identify (Cases et al. subsequently in fungus Mortierella (Mortierella), people and mice, J.Biol.Chem., 2001,276,38870-38876; Lardizabal et al., J.Biol.Chem., 2001,276,38862-38869).Enzymatic determination shows that this protein of identifying recently has the DG transferase active really, this activity can utilize multiple long-chain fat acyl group-CoA substrate (Cases et al., J.Biol.Chem., 2001,276,38870-38876).

DG transferring enzyme 2 is members of the family of the irrelevant gene of its sequence and DG transferring enzyme 1.Except compare with DG transferring enzyme 1 on sequence different; external test proves that also DG transferring enzyme 2 has higher activity (Cases et al., J.Biol.Chem., 2001 under the magnesium chloride of low concentration and oleoyl-CoA; 276,38870-38876).The predicted protein matter sequence of DG transferring enzyme 2 contains the potential N-of at least one membrane spaning domain of inferring, three and connects glycosylation site, six total sites of potential Protein kinase C phosphorylation and infer the common sequence in phosphoglycerol site (Cases et al. in the acyltransferase with being present in; J.Biol.Chem.; 2001; 276,38870-38876).International radiation hybridization collection of illustrative plates committee (International RadiationHybrid Mapping Consortium) navigates to chromosome 11q13.3 with people's DG transferring enzyme 2.

In people tissue, in liver and white adipose tissue, detect the DG transferring enzyme 2 of top level, in mammary gland, testis and peripheral blood leucocyte level lower (Cases et al., J.Biol.Chem., 2001,276,38870-38876).In people's tissue, detect two kinds of mRNA of 2.4 and 1.8 kilobase, and the main DG transferring enzyme 2mRNA in the mouse tissue is 2.4 kilobase.Except liver and white adipose tissue; DG transferring enzyme 2 is also expressed in all sections of mouse small intestine; wherein in near-end intestinal (proximalintestine), express higher; in distal gut (distal intstine), express lower (Cases etal.; J.Biol.Chem.; 2001,276,38870-38876).

In the postnatal development process of rat liver, the DG transferase active demonstrates unique pattern.Because mRNA expresses and active pattern between do not have dependency, post translational modification may participate in DG transferring enzyme 2 active adjustings in the rat growth course (Waterman et al., J.Lipid.Res., 2002,43,1555-1562).

DG transferring enzyme 2mRNA preferentially raises by insulinize; this active external test of DG (Meegalla et al. that is confirmed by the cell membrane part of the mice adipose cell of measure cultivating; Biochem.Biophys.Res.Commun.; 2002; 298,317-323).In the mice of fasting, DG transferring enzyme 2 is expressed and is greatly reduced, and sharply increases during feeding once more.These two kinds of acetyl-CoA carboxylase and fatty acid synthase participate in the synthetic enzyme of fatty acid, and its expression pattern responds fasting and feeding more in a similar manner.These results are in conjunction with this observed result of DG transferring enzyme 2 expressed in abundance in liver; prompting DG transferring enzyme 2 and endogenous fatty acid route of synthesis be closely connected (Meegalla etal., Biochem.Biophys.Res.Commun., 2002; 298,317-323).

Provide such evidence to the research that has destructive mice in diacylglycerol acyltransferase 1 gene, promptly diacylglycerol acyltransferase 2 helps triglyceride synthetic.Intestinal in wild-type mice and DG transferring enzyme 1 deficient mice is in each stage, DG transferring enzyme 2mRNA expression levels similar (Buhman et al., J.Biol.Chem., 2002,277,25474-25479).Use magnesium chlorides such as Buhman are distinguished DG transferring enzyme 1 and 2 activity; observe in DG transferring enzyme 1 deficient mice; the DG transferase active is reduced to 50% in the near-end intestinal; in distal gut, be reduced to 10-15% (Buhman et al.; J.Biol.Chem.; 2002,277,25474-25479).

In addition, in the liver or fatty tissue of DG transferring enzyme 1 deficient mice, even behind the high fat diet in several weeks, DG transferring enzyme 2mRNA level is not raised (Cases et al., J.Biol.Chem., 2001 yet, 276,38870-38876; Chen etal., J.Clin.Invest., 2002,109,1049-1055).But; in ob/ob mice (it has the sudden change that can cause fat disease in the leptin gene); DG transferring enzyme 2 is than there being the more expression of height in wild-type mice, this prompting DG transferring enzyme 2 may cause the liparitosis that existing height accumulates in these mices on the part.In addition; the combinatorial mutagenesis of leptin and DG transferring enzyme 1; the level that can cause DG transferring enzyme 2 in the white adipose tissue to be expressed in the homologue with DG transferring enzyme 1 deficient mice is compared three times of (the Chen et al. that raise; J.Clin.Invest.; 2002; 109,1049-1055).DG transferring enzyme 2mRNA also in the skin of these mices, obtain raising (Chen et al., J.Clin.Invest., 2002,109,175-181).These Notes of Key Datas; leptin can be reduced DG transferring enzyme 2 usually and express; and the rise of DG transferring enzyme 2 in the white adipose tissue of these mices can give the triglyceride that still appears in leptin deficiency/DG transferring enzyme 1 deficient mice synthetic approach (the Chen et al. that alternative is provided; J.Clin.Invest.; 2002; 109,1049-1055).

Diacylglycerol acyltransferase 1 is rejected mice and is demonstrated attractive Phenotype, and promptly they are very lean, can resist the inductive obesity of meals, and the tissue triglycerides level reduces, to the sensitivity raising of insulin and leptin.Because it is synthetic that DG transferring enzyme 2 also participates in triglyceride, also can cause body fat content to reduce similarly to the interference of DG transferring enzyme 2.

Definition

" DGAT2 " means its expression and treats by lacking gene outcome or the protein that antisense compounds is regulated.

" DGAT2 nucleic acid " means the nucleic acid of any encoding D GAT2.For example, RNA sequence that in certain embodiments, DGAT2 nucleic acid includes but not limited to the DNA sequence of encoding D GAT2, transcribe from the DNA of encoding D GAT2 and the mRNA sequence of encoding D GAT2." DGAT2 mRNA " means the mRNA of encoding D GAT2.

The treatment indication

Antisense technology is to reduce the effective means that DGAT2 expresses, and has been proved to be in multiple therapeutic, diagnosis type and the application of research property to have unique applications.Thus, in certain embodiments, the invention provides targeting encoding D GAT2 nucleic acid, can regulate the short antisense compounds of the expression of DGAT2.This paper provides in addition and can effectively suppress the short antisense compounds that DGAT2 expresses.

In certain embodiments, the short antisense compounds of the nucleic acid by giving one or more targeting encoding D GAT2 is treated and is suspected the experimenter who suffers from DGAT2 diseases associated or disease.For example, in non-limiting embodiments, this abundant short antisense compounds of will treat effective dose that comprises gives the step of this animal.In some this embodiment, short antisense compounds can effectively suppress the active of DGAT2 or suppress the expression of DGAT2.In one embodiment, the active or expression of DGAT2 in the experimenter is suppressed at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% and at least 100%.In certain embodiments, the active or expression of DGAT2 in the experimenter is suppressed about 30%.More preferably, the active or expression of DGAT2 in the experimenter is suppressed 50% or more.

The reduction of the expression of DGAT2 can for example be measured in blood, blood plasma, serum, fatty tissue, liver or any other body fluid, tissue or the organ of this animal.Preferably, the central contained cell of this analyzed fluid, tissue or organ contains nucleic acid and/or the DGAT2 albumen itself of encoding D GAT2.

In certain embodiments, also provide medicine and other compositionss that comprise The compounds of this invention.For example, the short antisense compounds of targeting DGAT2 nucleic acid can be added to suitable medicine by certain chemical compound with effective dose can accept diluent or carrier, uses in pharmaceutical composition.

The short antisense compounds of some targeting DGAT2 can have short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-10-1,2-10-2 and 3-10-3.

This paper provides by short antisense compounds that gives one or more targeting DGAT2 nucleic acid or the pharmaceutical composition that comprises this chemical compound and treats individual method.Also provide by the short antisense compounds that gives targeting DGAT2 nucleic acid and treat the method for suffering from the experimenter of active diseases associated of DGAT2 or disease.Include but not limited to cardiovascular disease, obesity, diabetes, hypercholesterolemia and liver steatosis with DGAT2 diseases associated or disease.

The short antisense compounds of some targeting DGAT2 nucleic acid

In certain embodiments, lacking the antisense compounds targeting has

The DGAT2 nucleic acid of the sequence of searching number NM_032564.2 (being incorporated herein) with SEQ ID NO:10.

In some this embodiment, short antisense compounds and the SEQID NO:10 of targeting SEQ ID NO:10 have at least 90% complementarity.In some this embodiment, the short antisense compounds of targeting SEQ IDNO:10 and SEQ ID NO:10 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ IDNO:10 of targeting SEQ ID NO:10 have 100% complementarity.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:10 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 14 and 15.

The nucleotide sequence that each SEQ ID NO provides in the table 14 and 15 is independent of any to connecting the modification of key or nuclear base between sugar moieties, nucleoside.Thus, the short antisense compounds that comprises the nucleotide sequence that provides in table 14 and 15 can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 14 and 15 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:10.Table 14 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:10.Table 15 has been enumerated the short antisense compounds that has one or two mispairing with respect to SEQ ID NO:10.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2 MOE " means 2-10-2 gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Table 14: the short antisense compounds of targeting SEQ ID NO:10

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						372556	231	244	ATGAGGGTCTTCAT	2-10-2 MOE	681
372557	249	262	ACCCCGGAGTAGGC	2-10-2 MOE	682
						382601	249	260	CCCGGAGTAGGC	1-10-1 MOE	683
372480	251	266	CAGGACCCCGGAGTAG	3-10-3 MOE	684
						372481	252	267	GCAGGACCCCGGAGTA	3-10-3 MOE	685
372558	252	265	AGGACCCCGGAGTA	2-10-2 MOE	686
						372559	253	266	CAGGACCCCGGAGT	2-10-2 MOE	687
382603	331	342	CAGACCCCTCGC	1-10-1 MOE	688
						382604	361	372	AGAGGATGCTGG	1-10-1 MOE	689
372485	392	407	GAGCCAGGTGACAGAG	3-10-3 MOE	690
						372563	393	406	AGCCAGGTGACAGA	2-10-2 MOE	691
382605	397	408	TGAGCCAGGTGA	1-10-1 MOE	692
						372565	414	427	TTTTCCACCTTGGA	2-10-2 MOE	693
382606	482	493	CTGCAGGCCACT	1-10-1 MOE	694
						372497	651	666	TCACCAGCTGGATGGG	3-10-3 MOE	695
372498	652	667	TTCACCAGCTGGATGG	3-10-3 MOE	696
						372575	652	665	CACCAGCTGGATGG	2-10-2 MOE	697
372576	653	666	TCACCAGCTGGATG	2-10-2 MOE	698
						382607	655	666	TCACCAGCTGGA	1-10-1 MOE	699
372499	656	671	TGTCTTCACCAGCTGG	3-10-3 MOE	700
						372577	657	670	GTCTTCACCAGCTG	2-10-2 MOE	701
372500	659	674	GTGTGTCTTCACCAGC	3-10-3 MOE	702
						372578	660	673	TGTGTCTTCACCAG	2-10-2 MOE	703
372501	661	676	TTGTGTGTCTTCACCA	3-10-3 MOE	704
						372579	662	675	TGTGTGTCTTCACC	2-10-2 MOE	705
372502	664	679	AGGTTGTGTGTCTTCA	3-10-3 MOE	706
						372580	665	678	GGTTGTGTGTCTTC	2-10-2 MOE	707
372503	666	681	GCAGGTTGTGTGTCTT	3-10-3 MOE	708
						372581	667	680	CAGGTTGTGTGTCT	2-10-2 MOE	709
372504	669	684	TCAGCAGGTTGTGTGT	3-10-3 MOE	710
						372582	670	683	CAGCAGGTTGTGTG	2-10-2 MOE	711
372505	671	686	GGTCAGCAGGTTGTGT	3-10-3 MOE	712
						372506	672	687	TGGTCAGCAGGTTGTG	3-10-3 MOE	713
372583	672	685	GTCAGCAGGTTGTG	2-10-2 MOE	714
						372584	673	686	GGTCAGCAGGTTGT	2-10-2 MOE	715
372507	676	691	CTGGTGGTCAGCAGGT	3-10-3 MOE	716
						372585	677	690	TGGTGGTCAGCAGG	2-10-2 MOE	717
382608	680	691	CTGGTGGTCAGC	1-10-1 MOE	718
						372508	681	696	AGTTCCTGGTGGTCAG	3-10-3 MOE	719
372586	682	695	GTTCCTGGTGGTCA	2-10-2 MOE	720
						372509	684	699	TATAGTTCCTGGTGGT	3-10-3 MOE	721
372587	685	698	ATAGTTCCTGGTGG	2-10-2 MOE	722
						372510	686	701	GATATAGTTCCTGGTG	3-10-3 MOE	723
372588	687	700	ATATAGTTCCTGGT	2-10-2 MOE	724
						372511	691	706	CCAAAGATATAGTTCC	3-10-3 MOE	725
372512	692	707	TCCAAAGATATAGTTC	3-10-3 MOE	726
						372589	692	705	CAAAGATATAGTTC	2-10-2 MOE	727

372590	693	706	CCAAAGATATAGTT	2-10-2 MOE	728
						382609	724	735	CCAGGCCCATGA	1-10-1 MOE	729
372514	725	740	GGCACCCAGGCCCATG	3-10-3 MOE	730
						372592	726	739	GCACCCAGGCCCAT	2-10-2 MOE	731
372515	730	745	CAGAAGGCACCCAGGC	3-10-3 MOE	732
						372593	731	744	AGAAGGCACCCAGG	2-10-2 MOE	733
382610	851	862	CCAGACATCAGG	1-10-1 MOE	734
						382611	867	878	GACAGGGCAGAT	1-10-1 MOE	735
382602	868	879	TGACAGGGCAGA	1-10-1 MOE	736
						382612	911	922	CCACTCCCATTC	1-10-1 MOE	737
372524	965	980	GCCAGGCATGGAGCTC	3-10-3 MOE	738
						372602	966	979	CCAGGCATGGAGCT	2-10-2 MOE	739
382613	968	979	CCAGGCATGGAG	1-10-1 MOE	740
						382614	987	998	CAGGGTGACTGC	1-10-1 MOE	741
372525	989	1004	GTTCCGCAGGGTGACT	3-10-3 MOE	742
						372603	990	1003	TTCCGCAGGGTGAC	2-10-2 MOE	743
372526	992	1007	GCGGTTCCGCAGGGTG	3-10-3 MOE	744
						372604	993	1006	CGGTTCCGCAGGGT	2-10-2 MOE	745
372530	1106	1121	TCGGCCCCAGGAGCCC	3-10-3 MOE	746
						372608	1107	1120	CGGCCCCAGGAGCC	2-10-2 MOE	747
372531	1109	1124	CCATCGGCCCCAGGAG	3-10-3 MOE	748
						372609	1110	1123	CATCGGCCCCAGGA	2-10-2 MOE	749
372532	1112	1127	GACCCATCGGCCCCAG	3-10-3 MOE	750
						372610	1113	1126	ACCCATCGGCCCCA	2-10-2 MOE	751
372533	1117	1132	TTCTGGACCCATCGGC	3-10-3 MOE	752
						382615	1117	1128	GGACCCATCGGC	1-10-1 MOE	753
372611	1118	1131	TCTGGACCCATCGG	2-10-2 MOE	754
						372536	1199	1214	CACCAGCCCCCAGGTG	3-10-3 MOE	755
372614	1200	1213	ACCAGCCCCCAGGT	2-10-2 MOE	756
						372537	1204	1219	TAGGGCACCAGCCCCC	3-10-3 MOE	757
372615	1205	1218	AGGGCACCAGCCCC	2-10-2 MOE	758
						372538	1209	1224	TGGAGTAGGGCACCAG	3-10-3 MOE	759
372616	1210	1223	GGAGTAGGGCACCA	2-10-2 MOE	760
						382616	1215	1226	CTTGGAGTAGGG	1-10-1 MOE	761
372539	1218	1233	TGATGGGCTTGGAGTA	3-10-3 MOE	762
						372617	1219	1232	GATGGGCTTGGAGT	2-10-2 MOE	763
372540	1293	1308	TGTGGTACAGGTCGAT	3-10-3 MOE	764
						372618	1294	1307	GTGGTACAGGTCGA	2-10-2 MOE	765
382617	1294	1305	GGTACAGGTCGA	1-10-1 MOE	766
						372541	1295	1310	GGTGTGGTACAGGTCG	3-10-3 MOE	767
372619	1296	1309	GTGTGGTACAGGTC	2-10-2 MOE	768
						372542	1298	1313	CATGGTGTGGTACAGG	3-10-3 MOE	769
372620	1299	1312	ATGGTGTGGTACAG	2-10-2 MOE	770
						372543	1300	1315	TACATGGTGTGGTACA	3-10-3 MOE	771
372621	1301	1314	ACATGGTGTGGTAC	2-10-2 MOE	772
						372544	1303	1318	ATGTACATGGTGTGGT	3-10-3 MOE	773
372622	1304	1317	TGTACATGGTGTGG	2-10-2 MOE	774
						382618	1313	1324	GCCTCCATGTAC	1-10-1 MOE	775
382619	1325	1336	AGCTTCACCAGG	1-10-1 MOE	776
						382620	1383	1394	GTTCACCTCCAG	1-10-1 MOE	777

Table 15: targeting SEQ ID NO:10 and have the short antisense compounds of 1 or 2 mispairing

ISIS NO	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						372608	151	164	CGGCCCCAGGAGCC	2-10-2 MOE	747
372474	156	171	CATGCCCCAGCCGCCG	3-10-3 MOE	778
						372552	157	170	ATGCCCCAGCCGCC	2-10-2 MOE	779
382609	167	178	CCAGGCCCATGA	1-10-1 MOE	729
						372478	230	245	GATGAGGGTCTTCATG	3-10-3 MOE	780
372479	248	263	GACCCCGGAGTAGGCA	3-10-3 MOE	781
						382611	317	328	GACAGGGCAGAT	1-10-1 MOE	735
372483	352	367	ATGCTGGAGCCAGTGC	3-10-3 MOE	782
						372561	353	366	TGCTGGAGCCAGTG	2-10-2 MOE	783
372562	373	386	GTCTTGGAGGGCCG	2-10-2 MOE	784
						382602	388	399	TGACAGGGCAGA	1-10-1 MOE	736
372613	392	405	CCCAGGTGTCAGAG	2-10-2 MOE	785
						372486	412	427	TTTTCCACCTTGGATC	3-10-3 MOE	786
372564	413	426	TTTCCACCTTGGAT	2-10-2 MOE	787
						372487	413	428	TTTTTCCACCTTGGAT	3-10-3 MOE	788
372488	418	433	AGGTGTTTTTCCACCT	3-10-3 MOE	789
						372566	419	432	GGTGTTTTTCCACC	2-10-2 MOE	790
372489	459	474	CCAGGAAGGATAGGAC	3-10-3 MOE	791
						372567	460	473	CAGGAAGGATAGGA	2-10-2 MOE	792
382612	475	486	CCACTCCCATTC	1-10-1 MOE	737
						372490	483	498	TGACACTGCAGGCCAC	3-10-3 MOE	793
372568	484	497	GACACTGCAGGCCA	2-10-2 MOE	794
						372491	492	507	ACATGAGGATGACACT	3-10-3 MOE	795
372569	493	506	CATGAGGATGACAC	2-10-2 MOE	796
						372492	503	518	GCAGAAGGTGTACATG	3-10-3 MOE	797
372570	504	517	CAGAAGGTGTACAT	2-10-2 MOE	798
						372493	512	527	GCAGTCAGTGCAGAAG	3-10-3 MOE	799
372571	513	526	CAGTCAGTGCAGAA	2-10-2 MOE	800
						372496	612	627	ACACGGCCCAGTTTCG	3-10-3 MOE	801
372574	613	626	CACGGCCCAGTTTC	2-10-2 MOE	802
						372513	717	732	GGCCCATGATGCCATG	3-10-3 MOE	803
372591	718	731	GCCCATGATGCCAT	2-10-2 MOE	804
						372516	732	747	TACAGAAGGCACCCAG	3-10-3 MOE	805
372594	733	746	ACAGAAGGCACCCA	2-10-2 MOE	806
						372518	812	827	GAAGTTGCCAGCCAAT	3-10-3 MOE	807
372596	813	826	AAGTTGCCAGCCAA	2-10-2 MOE	808
						372560	863	876	CAGGGCAGATCCTT	2-10-2 MOE	809
372519	887	902	CAAGTAGTCTATGGTG	3-10-3 MOE	810
						372597	888	901	AAGTAGTCTATGGT	2-10-2 MOE	811
372520	894	909	TGGAAAGCAAGTAGTC	3-10-3 MOE	812
						372598	895	908	GGAAAGCAAGTAGT	2-10-2 MOE	813
372527	1013	1028	GGCCAGCTTTACAAAG	3-10-3 MOE	814
						372605	1014	1027	GCCAGCTTTACAAA	2-10-2 MOE	815
372606	1020	1033	CGCAGGGCCAGCTT	2-10-2 MOE	816
						372529	1052	1067	AAAGGAATAGGTGGGA	3-10-3 MOE	817

372607	1053	1066	AAGGAATAGGTGGG	2-10-2 MOE	818
						372534	1144	1159	GCGAAACCAATATACT	3-10-3 MOE	819
372612	1145	1158	CGAAACCAATATAC	2-10-2 MOE	820
						372535	1192	1207	CCCCAGGTGTCAGAGG	3-10-3 MOE	821
372613	1193	1206	CCCAGGTGTCAGAG	2-10-2 MOE	822
						372545	1332	1347	GATTGTCAAAGAGCTT	3-10-3 MOE	823
372623	1333	1346	ATTGTCAAAGAGCT	2-10-2 MOE	824
						372546	1342	1357	TTGGTCTTGTGATTGT	3-10-3 MOE	825
372624	1343	1356	TGGTCTTGTGATTG	2-10-2 MOE	826
						372547	1352	1367	AAGGCCGAATTTGGTC	3-10-3 MOE	827
372625	1353	1366	AGGCCGAATTTGGT	2-10-2 MOE	828
						382601	1617	1628	CCCGGAGTAGGC	1-10-1 MOE	683
382606	1971	1982	CTGCAGGCCACT	1-10-1 MOE	694
						382612	1988	1999	CCACTCCCATTC	1-10-1 MOE	737

In certain embodiments, the target zone is the nucleotide 231-267 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 231-267 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 231-267 comprises the nucleotide sequence that is selected from SEQ ID NO 681,682,683,684,685,686 or 687.In some this embodiment, the short antisense compounds of the nucleotide 231-267 of targeting SEQ ID NO:10 is selected from Isis No 372556,372557,382601,372480,372481,372558 or 372559.

In certain embodiments, the target zone is the nucleotide 249-267 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 249-267 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 249-267 comprises the nucleotide sequence that is selected from SEQ ID NO 683,684,685,686 or 687.In some this embodiment, the short antisense compounds of the nucleotide 249-267 of targeting SEQ ID NO:10 is selected from Isis No 382601,372480,372481,372558 or 372559.

In certain embodiments, the target zone is the nucleotide 331-493 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 331-493 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 331-493 comprises the nucleotide sequence that is selected from SEQ ID NO 688,689,690,691,692,693 or 694.In some this embodiment, the short antisense compounds of the nucleotide 331-493 of targeting SEQ ID NO:10 is selected from Isis No 382603,382604,372485,372563,382605,372565 or 382606.

In certain embodiments, the target zone is the nucleotide 331-427 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 331-427 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 331-427 comprises the nucleotide sequence that is selected from SEQ ID NO 688,689,690,691,692 or 693.In some this embodiment, the short antisense compounds of the nucleotide 331-427 of targeting SEQ ID NO:10 is selected from Isis No 382603,382604,372485,372563,382605 or 372565.

In certain embodiments, the target zone is the nucleotide 392-408 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 392-408 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 392-408 comprises the nucleotide sequence that is selected from SEQ ID NO 690,691 or 692.In some this embodiment, the short antisense compounds of the nucleotide 392-408 of targeting SEQ ID NO:10 is selected from Isis No 372485,372563 or 382605.

In certain embodiments, the target zone is the nucleotide 651-707 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 651-707 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 651-707 comprises the nucleotide sequence that is selected from SEQ ID NO 695,696,697,698,699,700,701,702,703,704,705,706,707,708,709,710,711,712,713,714,715,716,717,718,719,720,721,722,723,724,725,726,727 or 728.In some this embodiment, the short antisense compounds of the nucleotide 651-707 of targeting SEQ ID NO:10 is selected from Isis No372497,372498,372575,372576,382607,372499,372577,372500,372578,372501,372579,372502,372580,372503,372581,372504,372582,372505,372506,372583,372584,372507,372585,382608,372508,372586,372509,372587,372510,372588,372511,372512,372589 or 372590.

In certain embodiments, the target zone is the nucleotide 724-745 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 724-745 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 724-745 comprises the nucleotide sequence that is selected from SEQ ID NO 729,730,731,732 or 733.In some this embodiment, the short antisense compounds of the nucleotide 724-745 of targeting SEQ ID NO:10 is selected from Isis No 382609,372514,372592,372515 or 372593.

In certain embodiments, the target zone is the nucleotide 651-745 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 651-745 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 651-745 comprises the nucleotide sequence that is selected from SEQ ID NO 695,696,697,698,699,700,701,702,703,704,705,706,707,708,709,710,711,712,713,714,715,716,717,718,719,720,721,722,723,724,725,726,727,728,729,730,731,732 or 733.In some this embodiment, the short antisense compounds of the nucleotide 651-745 of targeting SEQ ID NO:10 is selected from Isis No 372497,372498,372575,372576,382607,372499,372577,372500,372578,372501,372579,372502,372580,372503,372581,372504,372582,372505,372506,372583,372584,372507,372585,382608,372508,372586,372509,372587,372510,372588,372511,372512,372589,372590,382609,372514,372592,372515 or 372593.

In certain embodiments, the target zone is the nucleotide 851-922 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 851-922 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 851-922 comprises the nucleotide sequence that is selected from SEQ ID NO 734,735,736 or 737.In some this embodiment, the short antisense compounds of the nucleotide 851-922 of targeting SEQ ID NO:10 is selected from Isis No 382610,382611,382602 or 382612.

In certain embodiments, the target zone is the nucleotide 851-879 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 851-879 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 851-879 comprises the nucleotide sequence that is selected from SEQ ID NO 734,735 or 736.In some this embodiment, the short antisense compounds of the nucleotide 851-879 of targeting SEQ ID NO:10 is selected from Isis No 382610,382611 or 382602.

In certain embodiments, the target zone is the nucleotide 965-1007 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 965-1007 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 965-1007 comprises the nucleotide sequence that is selected from SEQ ID NO 738,739,740,741,742,743,744 or 745.In some this embodiment, the short antisense compounds of the nucleotide 965-1007 of targeting SEQ ID NO:10 is selected from Isis No 372524,372602,382613,382614,372525,372603,372526 or 372604.

In certain embodiments, the target zone is the nucleotide 965-979 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 965-979 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 965-979 comprises the nucleotide sequence that is selected from SEQ ID NO 738,739 or 740.In some this embodiment, the short antisense compounds of the nucleotide 965-979 of targeting SEQ ID NO:10 is selected from Isis No 372524,372602 or 382613.

In certain embodiments, the target zone is the nucleotide 987-1007 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 987-1007 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 987-1007 comprises the nucleotide sequence that is selected from SEQ ID NO 741,742,743,744 or 745.In some this embodiment, the short antisense compounds of the nucleotide 987-1007 of targeting SEQ ID NO:10 is selected from Isis No 382614,372525,372603,372526 or 372604.

In certain embodiments, the target zone is the nucleotide 1106-1132 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 1106-1132 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 1106-1132 comprises the nucleotide sequence that is selected from SEQ ID NO 746,747,748,749,750,751,752,753 or 754.In some this embodiment, the short antisense compounds of the nucleotide 1106-1132 of targeting SEQ IDNO:10 is selected from Isis No 372530,372608,372531,372609,372532,372610,372533,382615 or 372611.

In certain embodiments, the target zone is the nucleotide 1199-1233 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 1199-1233 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 1199-1233 comprises the nucleotide sequence that is selected from SEQ ID NO 755,756,757,758,759,760,761,762 or 763.In some this embodiment, the short antisense compounds of the nucleotide 1199-1233 of targeting SEQ IDNO:10 is selected from Isis No 372536,372614,372537,372615,372538,372616,382616,372539 or 372617.

In certain embodiments, the target zone is the nucleotide 1293-1394 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 1293-1394 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 1293-1394 comprises the nucleotide sequence that is selected from SEQ ID NO 764,765,766,767,768,769,770,771,772,773,774,775,776 or 777.In some this embodiment, the short antisense compounds of the nucleotide 1293-1394 of targeting SEQ ID NO:10 is selected from Isis No 372540,372618,382617,372541,372619,372542,372620,372543,372621,372544,372622,382618,382619 or 382620.

In certain embodiments, the target zone is the nucleotide 1293-1336 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 1293-1336 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 1293-1336 comprises the nucleotide sequence that is selected from SEQ ID NO 764,765,766,767,768,769,770,771,772,773,774,775 or 776.In some this embodiment, the short antisense compounds of the nucleotide 1293-1336 of targeting SEQ ID NO:10 is selected from Isis No 372540,372618,382617,372541,372619,372542,372620,372543,372621,372544,372622,382618 or 382619.

In certain embodiments, the target zone is the nucleotide 1293-1324 of SEQ ID NO:10.In certain embodiments, lack the nucleotide 1293-1324 of antisense compounds targeting SEQ ID NO:10.In some this embodiment, the short antisense compounds of targeted nucleotide 1293-1324 comprises the nucleotide sequence that is selected from SEQ ID NO 764,765,766,767,768,769,770,771,772,773,774 or 775.In some this embodiment, the short antisense compounds of the nucleotide 1293-1324 of targeting SEQ ID NO:10 is selected from Isis No 372540,372618,382617,372541,372619,372542,372620,372543,372621,372544,372622 or 382618.

In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting DGAT2 nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 19 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting DGAT2 nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting DGAT2 nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating DGAT2.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting DGAT2 nucleic acid, and wherein the short antisense compounds of this targeting DGAT2 nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting DGAT2 nucleic acid is regulated the expression of DGAT2.

In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 8 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 9 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 10 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 11 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 12 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 13 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 14 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 15 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for adjusting DGAT2 comprises that using length is the short antisense compounds of 16 monomeric targeting DGAT2 nucleic acid.

In certain embodiments, the method for the expression of adjusting DGAT2 comprises that use comprises the short antisense compounds of 9-15 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for the expression of adjusting DGAT2 comprises that use comprises the short antisense compounds of 10-15 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for the expression of adjusting DGAT2 comprises that use comprises the short antisense compounds of 12-14 monomeric targeting DGAT2 nucleic acid.In certain embodiments, the method for the expression of adjusting DGAT2 comprises the short antisense compounds that uses the targeting DGAT2 nucleic acid that comprises 12-14 nucleotide or nucleoside.

9.PTP1B

PTP1B (also claiming protein phosphatase 1 B and PTPN1) is endoplasmic reticulum (ER) relevant enzyme, is as the main protein tyrosine phosphatase of people's Placenta Hominis separated (Tonks et al., J.Biol.Chem., 1988,263,6731-6737 at first; Tonks et al., J.Biol.Chem., 1988,263,6722-6730).

The clear and definite necessary regulating action of PTP1B in the Insulin receptor INSR Mediated Signal Transduction.In some cases, PTP1B is external and can both take place to interact and with its dephosphorylation, cause downward modulation (the Goldstein et al. of signal transduction pathway with activatory Insulin receptor INSR in intact cell, Mol.Cell.Biochem, 1998,182,91-99; Seely et al., Diabetes, 1996,45,1379-1385).In addition, PTP1B can regulate insulin the mitogenesis effect (Goldstein et al., Mol.Cell.Biochem, 1998,182,91-99).In the rat fat cell of overexpression PTP1B, the transposition of GLUT4 glucose transporter is suppressed, this hint PTP1B also be glucose transport negative growth factor (Chen etal., J.Biol.Chem., 1997,272,8026-8031).

The mice that lacks the PTP1B gene is rejected the negative adjusting that model shows that also PTP1B transduces to insulin signaling.The mice that has ruined PTP1B gene demonstrates insulin sensitivity increase and the increase of Insulin receptor INSR phosphorylation.PTP1B-/-mice when being given high fat diet, can resist weight increase and keep insulin sensitivity (Elchebly et al., Science, 1999,283,1544-1548).The obvious establishment of these researchs PTP1B is the treatment target in diabetes and the bariatrician.

Diabetes and obesity (being generically and collectively referred to as " sugared fat disease (diabesity) " now sometimes) are inter-related.Most of human obesity diseases are relevant with the leptin resistance with insulin resistance.In fact, obesity may than diabetes itself to the influence of insulin action bigger (Sindelka et al., Physiol Res., 2002,51,85-91).Syndrome X or metabolic syndrome are the new terminologies of such one group of disease, and this group disease can show the tendency that development diabetes and cardiovascular disease are arranged when occurring together.These symptoms comprise hypertension, high triglyceride, low HDL and obesity, tend to occur together in some individualities.Because the effect of PTP1B in diabetes and obesity, it has been considered to comprise the treatment target of the multiple metabolic disorder of diabetes, obesity and metabolic syndrome.By improving blood-glucose control, the inhibitor of PTP1B also can be used for slowing down, prevent, postponing or improve the sequela of diabetes, and these sequela comprise retinopathy, neuropathy, cardiovascular complication and nephropathy.

In the cell cycle process, be subjected to difference and regulate (Schievella et al., Cell.GrowthDiffer., 1993,4, PTP1B 239-246), in insulin sensitivity sex organization, be expressed as two kinds of different isotypes, these two kinds of isotypes are the generation of road montage (alternate splicing) in addition (Shifrin and Neel, J.Biol.Chem., 1993 by pre-mRNA, 268,25376-25384).In addition the ratio of the product of road montage is subjected to the influence of somatomedin such as insulin, variously be subjected to have nothing in common with each other in the inspection tissue (Sell and Reese, Mol.Genet.Metab., 1999,66,189-192).In these researchs, the level of each variant is relevant with the body fat percentage rate with plasma insulin concentration.Therefore these variants can be used as chronic hyperinsulinemia and type ii diabetes patient's biological marker.

Definition

" Protein Tyrosine Phosphatases 1B " is that its expression is treated by lacking gene outcome or the protein that antisense compounds is regulated.Protein Tyrosine Phosphatases 1B is commonly referred to PTP1B, but also can be described as Protein Tyrosine Phosphatases; PTPN1; RKPTP; Protein Tyrosine Phosphatases, non-receptor type 1.

" PTP1B nucleic acid " means the nucleic acid of any coding PTP1B.For example, in certain embodiments, PTP1B nucleic acid include but not limited to the to encode DNA sequence of PTP1B, the RNA sequence of transcribing from the DNA of coding PTP1B and the mRNA sequence of coding PTP1B.

" PTP1B mRNA " means the proteic mRNA of coding PTP1B.

The treatment indication

Antisense technology is to reduce the effective means that PTP1B expresses, and has been proved to be in multiple therapeutic, diagnosis type and the application of research property to have unique applications.Thus, in certain embodiments, the invention provides targeting coding PTP1B nucleic acid, can regulate the short antisense compounds of the expression of PTP1B.This paper provides in addition and can effectively suppress the short antisense compounds that PTP1B expresses.

In some treatment, the short antisense compounds of the nucleic acid by giving one or more targeting coding PTP1B is treated and is suspected to suffer from the disease that can treat by the expression of adjusting PTP1B or the experimenter of disease.For example, in non-limiting embodiments, described method comprises that the short antisense compounds with the treatment effective dose gives the step of animal.Short antisense compounds of the present invention can effectively suppress the active of PTP1B and/or suppress the expression of PTP1B.In one embodiment, the active or expression of the PTP1B among the experimenter is suppressed and reaches at least 10%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99% or 100%.In certain embodiments, the active or expression of the PTP1B among the experimenter is suppressed and reaches about 30%.In certain embodiments, the active or expression of the PTP1B among the experimenter is suppressed and reaches at least 50% or more.

The reduction of the expression of PTP1B can for example be measured in blood, blood plasma, serum, fatty tissue, liver or any other body fluid, tissue or the organ of this animal.Preferably, the central contained cell of described analyzed fluid, tissue or organ contains nucleic acid and/or the PTP1B albumen itself of the PTP1B that encodes.

Some medicine that comprises The compounds of this invention and other compositionss also are provided.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid is to be added to suitable medicine by certain chemical compound with effective dose can accept diluent or carrier, uses in pharmaceutical composition.

The short antisense compounds of targeting PTP1B nucleic acid can have short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3 and 1-9-2.

In certain embodiments, this paper provides by short antisense compounds that gives one or more targeting PTP1B nucleic acid or the pharmaceutical composition that comprises this chemical compound and treats individual method.Also provide by the short antisense compounds that gives targeting PTP1B nucleic acid and treat the method for suffering from the experimenter of active diseases associated of PTP1B or disease.Include but not limited to high blood-glucose or hyperglycemia, prediabetes, diabetes, type ii diabetes, metabolic syndrome, obesity and insulin resistance with PTP1B diseases associated and disease.Therefore, this paper provides by giving the short antisense compounds of targeting PTP1B nucleic acid, treats the method for high blood-glucose or hyperglycemia, prediabetes, diabetes, type ii diabetes, metabolic syndrome, obesity and insulin resistance.

In certain embodiments, the invention provides in order to give the experimenter, reduce the blood-glucose among this experimenter or prevent or postpone the compositions and the method for the outbreak of blood glucose levels rising among this experimenter by the short antisense inhibitor that PTP1B is expressed.

In certain embodiments, the invention provides, improve the insulin sensitivity among this experimenter or prevent or postpone the compositions and the method for the outbreak of insulin resistance among this experimenter in order to give the experimenter by the short antisense inhibitor that PTP1B is expressed.

In certain embodiments, the invention provides in order to give the experimenter, treat the metabolic disorder among this experimenter or prevent or postpone the compositions and the method for the outbreak of metabolic disorder among this experimenter by short antisense compounds with targeting PTP1B nucleic acid.This metabolic disorder can be that any and PTP1B express relevant metabolic disorder, includes but not limited to diabetes and obesity.The method that reduces obesity (adiposity) also is provided.The wherein method of the obesity (obesity) of metabolic rate raising for the treatment of also is provided.

In certain embodiments, the experimenter suffers from type ii diabetes.In certain embodiments, the experimenter demonstrates high HbA1c level.In certain embodiments, the HbA1c level be at least about 6%, at least about 7%, at least about 8%, at least about 9%, at least about 10% or at least about 11%.In preferred embodiments, HbA_1cLevel reduces to about 7% or be lower than about 7%.In certain embodiments, the experimenter demonstrates high Body Mass Index.In certain embodiments, high Body Mass Index is greater than 25kg/m²In certain embodiments, the experimenter demonstrates hyperglycemia or hypertension glucose level.In a specific embodiment, blood glucose levels is a fasting blood pressure glucose level.In certain embodiments, high fasting blood pressure glucose level is 130mg/dL.In certain embodiments, the experimenter demonstrates hyperglycemia or demonstrates the fasting blood glucose levels before the treatment beginning and is higher than about 130mg/dL, baseline HbA_1cLevel at least about 7% or Body Mass Index greater than 25kg/m²Or their any combination.

In certain embodiments, provide the method that reduces one or more this levels by the short antisense compounds that gives targeting PTP1B nucleic acid.For example, provide by short antisense compounds and given the experimenter, reduced fasting glucose level, HbA among the experimenter targeting PTP1B_1cThe method of level or Body Mass Index level or their any combination.The fasting glucose can be fasting blood-glucose, fasting serum glucose or fasting plasma glucose.In some embodiments, fasting plasma glucose levels reduces at least about 25mg/dL or at least about 10mg/dL.In certain embodiments, described experimenter can not reach normal glucose level under the therapeutic scheme situation of accepting glucose attenuating medicament such as insulin, sulfonylureas or metformin.

In certain embodiments, the invention provides the method that changes lipid level.Some this method is to give the experimenter by the short antisense compounds with targeting PTP1B nucleic acid, reduces cholesterol, LDL and/or VLDL level among this experimenter or their any combination.In certain embodiments, the HDL level among the experimenter is to give this experimenter by the short antisense compounds with targeting PTP1B nucleic acid to improve.In certain embodiments, the LDL among the experimenter: HDL ratio and/or T-CHOL: HDL ratio are to give this experimenter by the short antisense compounds with targeting PTP1B nucleic acid to reduce.In certain embodiments, the HDL among the experimenter: LDL ratio and/or total HDL: the cholesterol ratio is to give this experimenter by the short antisense compounds with targeting PTP1B nucleic acid to improve.In certain embodiments, lipodogramme among the experimenter (lipid profile) is to give this experimenter by the short antisense compounds with targeting PTP1B nucleic acid, with raising HDL, reduction LDL, reduction VLDL, triglyceride reducing, reduction apolipoprotein B level or hypercholesterolemia reducing level or their combination, thereby be improved.In this embodiment, the experimenter is an animal, comprises the people.

Combination treatment

In certain embodiments, the pharmaceutical composition that one or more is comprised the short antisense compounds of targeting PTP1B nucleic acid gives altogether with one or more other medicament.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease identical with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease different with described one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament is designed to treat the ill effect of one or more pharmaceutical compositions of the present invention.In certain embodiments, one or more pharmaceutical compositions of the present invention are given altogether with another medicament, to treat the ill effect of this another medicament.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given simultaneously.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given at different time.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is prepared together in single dosage form.In certain embodiments, with other separately preparation of medicament of one or more pharmaceutical compositions of the present invention and one or more.

In certain embodiments, the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting PTP1B nucleic acid comprises that glucose reduces medicament and therapy.In some embodiments, glucose reduction medicament is PPAR agonist (gamma agonist, binary (dual) agonist or general (pan) agonist), dipeptidyl peptidase (IV) inhibitor, GLP-1 analog, insulin or insulin analog, insulin secretagogue, SGLT2 inhibitor, human branched chain starch analog, biguanide, alpha-glucosidase inhibitor, meglitinide, thiazolidinedione or sulfonylureas.

In some embodiments, glucose reduction medicament is the GLP-1 analog.In some embodiments, the GLP-1 analog is Exendin-4 or Li Lalu peptide (liraglutide).

In other embodiments, glucose reduction medicament is a sulfonylureas.In some embodiments, sulfonylureas is acetohexamide, chlorpropamide, tolbutamide, tolazamide, glimepiride, glipizide, glibenclamide or gliclazide.

In some embodiments, glucose reduction medicament is a biguanide.In some embodiments, biguanide is a metformin, and in some embodiments, reducing appears in the blood pressure glucose level, and lactic acidosis is as occurring increasing with the viewed lactic acidosis in metformin treatment back separately.

In some embodiments, glucose reduction medicament is a meglitinide.In some embodiments, meglitinide is Nateglinide or repaglinide.

In some embodiments, glucose reduction medicament is a thiazolidinedione.In some embodiments, thiazolidinedione is pioglitazone, rosiglitazone or troglitazone.In some embodiments, blood glucose levels occur to reduce, but weight increase do not resemble independent rosiglitazone treat viewed height.

In some embodiments, glucose reduction medicament is an alpha-glucosidase inhibitor.In some embodiments, alpha-glucosidase inhibitor is acarbose or miglitol.

In a certain embodiment, it is ISIS 113715 that the glucose that gives altogether reduces medicament.

In a certain embodiment, it is that curative life style changes that glucose reduces therapy.

In some this embodiment, glucose is reduced medicament give before the pharmaceutical composition of the present invention giving.In some this embodiment, glucose is reduced medicament after giving pharmaceutical composition of the present invention, give.In some this embodiment, glucose is reduced medicament when giving pharmaceutical composition of the present invention, give.In some this embodiment, the dosage that the glucose that gives altogether reduces medicament is identical with the dosage that gives separately to be given when glucose reduces medicament.In some this embodiment, the dosage that the glucose that gives altogether reduces medicament is lower than the dosage that is given when giving glucose reduction medicament separately.In some this embodiment, the dosage that the glucose that gives altogether reduces medicament is higher than the dosage that is given when giving glucose reduction medicament separately.

In certain embodiments, the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting PTP1B nucleic acid comprises that lipid reduces medicament.This lipid reduces medicament to be discussed in other places of the application, here is to be related to PTP1B and to be comprised.This lipid reduces medicament can reduce to glucose as above that medicament is described to be given.

In certain embodiments, the medicament that can give altogether with the pharmaceutical composition of the short antisense compounds that comprises targeting PTP1B nucleic acid comprises the anti-obesity medicament.This anti-obesity medicament can be as above reduces to glucose that medicament is described to be given.

Also provide the method that gives the short antisense compounds of targeting PTP1B nucleic acid by injection, this method also is included in the injection site and gives topical steroids.

Medicine

This paper also provides the short antisense compounds of targeting PTP1B nucleic acid to be used for preparation in order to reduce blood glucose levels (comprising the fasting glucose level) and HbA_1cThe purposes of the medicine of level, Body Mass Index level or their any combination.This medicine can give in load phase (loading period) and the phase of keeping.In some embodiments, this medicine gives by subcutaneous or intravenous.In other embodiments, described medicine be every day at least once, each week at least once or every month at least once.In a specific embodiment, the short antisense compounds that exists in this medicine is that particularly 20 or 20 dosage with the short antisense compounds of the sequence of coker base give to be lower than longer sequence.The experimenter that this medicine can be shown high blood-glucose or hyperglycemia, prediabetes, diabetes, type ii diabetes, metabolic syndrome, obesity and insulin resistance.

Other aspects and the advantage of short antisense compounds have introduction in this article.Disclosed herein, particularly for all aspects and the advantage of other targets, be applicable to the compositions of the short antisense compounds that comprises targeting PTP1B nucleic acid and their using method.

The short antisense compounds of some targeting PTP1B nucleic acid

In certain embodiments, lacking the antisense compounds targeting has

The sequence of searching number NM_002827.2 (being incorporated herein) with SEQ ID NO:11 or

The PTP1B nucleic acid of the nucleotide 14178000-1425600 of the sequence of searching number NT_011362.9 (being incorporated herein) with SEQ ID NO:12.In some this embodiment, short antisense compounds and the SEQ ID NO:11 of targeting SEQ ID NO:11 have at least 90% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:11 of targeting SEQ ID NO:11 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:11 of targeting SEQ ID NO:11 have 100% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:12 of targeting SEQ ID NO:12 have at least 90% complementarity.In some this embodiment, the short antisense compounds of targeting SEQID NO:12 and SEQ ID NO:12 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ IDNO:12 of targeting SEQ ID NO:12 have 100% complementarity.

In certain embodiments, the short antisense compounds of targeting SEQ ID NO:11 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 16 and 17.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:12 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 18 and 19.

Each nucleotide sequence that table 16,17,18 and 19 provides in each is independent of any modification to sugar moieties, Dan Julian key or nuclear base.Thus, the short antisense compounds that comprises certain nucleotide sequence that provides in the table 16,17,18 and 19 can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 16 and 17 has been enumerated the example of the short antisense compounds of targeting SEQ ID NO:11.Table 16 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:11.Table 17 has been enumerated the short antisense compounds that has one or two mispairing with respect to SEQ ID NO:11.Table 18 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:12.Table 19 has been enumerated the short antisense compounds that has 1 or 2 mispairing with respect to SEQ ID NO:12.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2MOE " means the 2-10-2gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Table 16: the short antisense compounds of targeting SEQ ID NO:11

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						147022	177	188	TTGTCGATCTCC	1-10-1MOE	886
147023	178	189	CTTGTCGATCTC	1-10-1MOE	859
						147024	179	190	CCTTGTCGATCT	1-10-1MOE	853
147019	195	206	TCGATCTCCTCG	1-10-1MOE	877
						147020	196	207	GTCGATCTCCTC	1-10-1MOE	868
147021	197	208	TGTCGATCTCCT	1-10-1MOE	882
						147022	198	209	TTGTCGATCTCC	1-10-1MOE	886
147023	199	210	CTTGTCGATCTC	1-10-1MOE	859
						147024	200	211	CCTTGTCGATCT	1-10-1MOE	853
147025	201	212	GCCTTGTCGATC	1-10-1MOE	865
						147026	202	213	AGCCTTGTCGAT	1-10-1MOE	835
147027	203	214	CAGCCTTGTCGA	1-10-1MOE	843
						147028	204	215	CCAGCCTTGTCG	1-10-1MOE	846
147073	204	215	CACTGATCCTGC	1-10-1MOE	842
						147029	205	216	CCCAGCCTTGTC	1-10-1MOE	848
147030	206	217	TCCCAGCCTTGT	1-10-1MOE	874
						147036	212	223	CCCAGTTCCCAG	1-10-1MOE	849
147037	213	224	GCCCAGTTCCCA	1-10-1MOE	863
						147038	214	225	CGCCCAGTTCCC	1-10-1MOE	855
147039	215	226	CCGCCCAGTTCC	1-10-1MOE	850
						147040	216	227	GCCGCCCAGTTC	1-10-1MOE	864
147041	217	228	AGCCGCCCAGTT	1-10-1MOE	834

147073	311	322	CACTGATCCTGC	1-10-1MOE	842
						147042	323	334	GGTCAAAAGGGC	1-10-1MOE	866
147043	324	335	TGGTCAAAAGGG	1-10-1MOE	881
						147044	325	336	GTGGTCAAAAGG	1-10-1MOE	869
147045	326	337	TGTGGTCAAAAG	1-10-1MOE	883
						147046	327	338	CTGTGGTCAAAA	1-10-1MOE	858
147047	328	339	ACTGTGGTCAAA	1-10-1MOE	833
						147051	332	343	TCCGACTGTGGT	1-10-1MOE	875
147052	333	344	ATCCGACTGTGG	1-10-1MOE	837
						147053	334	345	AATCCGACTGTG	1-10-1MOE	829
147054	335	346	TAATCCGACTGT	1-10-1MOE	871
						147055	336	347	TTAATCCGACTG	1-10-1MOE	884
147056	337	348	TTTAATCCGACT	1-10-1MOE	887
						147057	338	349	ATTTAATCCGAC	1-10-1MOE	839
147058	339	350	AATTTAATCCGA	1-10-1MOE	830
						147059	340	351	CAATTTAATCCG	1-10-1MOE	840
147060	341	352	GCAATTTAATCC	1-10-1MOE	861
						147061	342	353	TGCAATTTAATC	1-10-1MOE	879
147045	679	690	TGTGGTCAAAAG	1-10-1MOE	883
						147046	680	691	CTGTGGTCAAAA	1-10-1MOE	858
147045	787	798	TGTGGTCAAAAG	1-10-1MOE	883
						147046	788	799	CTGTGGTCAAAA	1-10-1MOE	858
147066	816	827	CCTGCACTGACG	1-10-1MOE	851
						404131	992	1005	ACCTTCGATCACAG	2-10-2MOE	831
147062	1024	1035	CACTGACGAGTC	1-10-1MOE	841
						147063	1025	1036	GCACTGACGAGT	1-10-1MOE	862
147064	1026	1037	TGCACTGACGAG	1-10-1MOE	880
						147065	1027	1038	CTGCACTGACGA	1-10-1MOE	857
147066	1028	1039	CCTGCACTGACG	1-10-1MOE	851
						147067	1029	1040	TCCTGCACTGAC	1-10-1MOE	876
147068	1030	1041	ATCCTGCACTGA	1-10-1MOE	838
						147069	1031	1042	GATCCTGCACTG	1-10-1MOE	860
147070	1032	1043	TGATCCTGCACT	1-10-1MOE	878
						147071	1033	1044	CTGATCCTGCAC	1-10-1MOE	856
147072	1034	1045	ACTGATCCTGCA	1-10-1MOE	832
						147073	1035	1046	CACTGATCCTGC	1-10-1MOE	842
147067	1199	1210	TCCTGCACTGAC	1-10-1MOE	876
						147040	1288	1299	GCCGCCCAGTTC	1-10-1MOE	864
147040	1396	1407	GCCGCCCAGTTC	1-10-1MOE	864
						147022	1868	1879	TTGTCGATCTCC	1-10-1MOE	886
147023	1869	1880	CTTGTCGATCTC	1-10-1MOE	859
						147024	1870	1881	CCTTGTCGATCT	1-10-1MOE	853
147019	1886	1897	TCGATCTCCTCG	1-10-1MOE	877
						147020	1887	1898	GTCGATCTCCTC	1-10-1MOE	868
147021	1888	1899	TGTCGATCTCCT	1-10-1MOE	882
						147022	1889	1900	TTGTCGATCTCC	1-10-1MOE	886
147023	1890	1901	CTTGTCGATCTC	1-10-1MOE	859
						147025	1892	1903	GCCTTGTCGATC	1-10-1MOE	865
147027	1894	1905	CAGCCTTGTCGA	1-10-1MOE	843
						147028	1895	1906	CCAGCCTTGTCG	1-10-1MOE	846
147030	1897	1908	TCCCAGCCTTGT	1-10-1MOE	874

147037	1904	1915	GCCCAGTTCCCA	1-10-1MOE	863
						147038	1905	1916	CGCCCAGTTCCC	1-10-1MOE	855
147040	1907	1918	GCCGCCCAGTTC	1-10-1MOE	864
						147041	1908	1919	AGCCGCCCAGTT	1-10-1MOE	834
147022	1976	1987	TTGTCGATCTCC	1-10-1MOE	886
						147023	1977	1988	CTTGTCGATCTC	1-10-1MOE	859
147024	1978	1989	CCTTGTCGATCT	1-10-1MOE	853
						147020	1995	2006	GTCGATCTCCTC	1-10-1MOE	868
147021	1996	2007	TGTCGATCTCCT	1-10-1MOE	882
						147022	1997	2008	TTGTCGATCTCC	1-10-1MOE	886
147023	1998	2009	CTTGTCGATCTC	1-10-1MOE	859
						147024	1999	2010	CCTTGTCGATCT	1-10-1MOE	853
147025	2000	2011	GCCTTGTCGATC	1-10-1MOE	865
						147026	2001	2012	AGCCTTGTCGAT	1-10-1MOE	835
147027	2002	2013	CAGCCTTGTCGA	1-10-1MOE	843
						147028	2003	2014	CCAGCCTTGTCG	1-10-1MOE	846
147029	2004	2015	CCCAGCCTTGTC	1-10-1MOE	848
						147030	2005	2016	TCCCAGCCTTGT	1-10-1MOE	874
147036	2011	2022	CCCAGTTCCCAG	1-10-1MOE	849
						147037	2012	2023	GCCCAGTTCCCA	1-10-1MOE	863
147038	2013	2024	CGCCCAGTTCCC	1-10-1MOE	855
						147039	2014	2025	CCGCCCAGTTCC	1-10-1MOE	850
147040	2015	2026	GCCGCCCAGTTC	1-10-1MOE	864
						147041	2016	2027	AGCCGCCCAGTT	1-10-1MOE	834
404199	2366	2379	GGTCATGCACAGGC	2-10-2MOE	867
						404134	2369	2382	TCAGGTCATGCACA	2-10-2MOE	873
404132	2548	2561	CCTTGGAATGTCTG	2-10-2MOE	852
						147020	2613	2624	GTCGATCTCCTC	1-10-1MOE	868
147020	2721	2732	GTCGATCTCCTC	1-10-1MOE	868
						404133	3289	3302	TATTCCATGGCCAT	2-10-2MOE	872
147032	6220	6231	GTTCCCAGCCTT	1-10-1MOE	870
						147033	6221	6232	AGTTCCCAGCCT	1-10-1MOE	836
147034	6222	6233	CAGTTCCCAGCC	1-10-1MOE	844
						147044	6288	6299	GTGGTCAAAAGG	1-10-1MOE	869
147045	6289	6300	TGTGGTCAAAAG	1-10-1MOE	883
						147032	6329	6340	GTTCCCAGCCTT	1-10-1MOE	870
147033	6330	6341	AGTTCCCAGCCT	1-10-1MOE	836
						147034	6331	6342	CAGTTCCCAGCC	1-10-1MOE	844
147044	6397	6408	GTGGTCAAAAGG	1-10-1MOE	869
						147045	6398	6409	TGTGGTCAAAAG	1-10-1MOE	883
147058	7057	7068	AATTTAATCCGA	1-10-1MOE	830
						147059	7058	7069	CAATTTAATCCG	1-10-1MOE	840
147060	7059	7070	GCAATTTAATCC	1-10-1MOE	861
						147058	7166	7177	AATTTAATCCGA	1-10-1MOE	830
147059	7167	7178	CAATTTAATCCG	1-10-1MOE	840
						147041	8084	8095	AGCCGCCCAGTT	1-10-1MOE	834
147041	8192	8203	AGCCGCCCAGTT	1-10-1MOE	834
						147027	8630	8641	CAGCCTTGTCGA	1-10-1MOE	843
147028	8631	8642	CCAGCCTTGTCG	1-10-1MOE	846
						147027	8738	8749	CAGCCTTGTCGA	1-10-1MOE	843
147028	8739	8750	CCAGCCTTGTCG	1-10-1MOE	846

147043	10957	10968	TGGTCAAAAGGG	1-10-1MOE	881
						147044	10958	10969	GTGGTCAAAAGG	1-10-1MOE	869
147043	11065	11076	TGGTCAAAAGGG	1-10-1MOE	881
						147044	11066	11077	GTGGTCAAAAGG	1-10-1MOE	869
147071	11605	11616	CTGA TCCTGCAC	1-10-1MOE	856
						147070	11611	11622	TGATCCTGCACT	1-10-1MOE	878
147071	11612	11623	CTGATCCTGCAC	1-10-1MOE	856
						147072	12294	12305	ACTGATCCTGCA	1-10-1MOE	832
147072	12299	12310	ACTGATCCTGCA	1-10-1MOE	832
						147030	12805	12816	TCCCAGCCTTGT	1-10-1MOE	874
147031	12806	12817	TTCCCAGCCTTG	1-10-1MOE	885
						147053	12939	12950	AATCCGACTGTG	1-10-1MOE	829
147030	12986	12997	TCCCAGCCTTGT	1-10-1MOE	874
						147031	12987	12998	TTCCCAGCCTTG	1-10-1MOE	885
147053	13120	13131	AATCCGACTGTG	1-10-1MOE	829
						147051	13162	13173	TCCGACTGTGGT	1-10-1MOE	875
147061	13316	13327	TGCAATTTAATC	1-10-1MOE	879
						147047	13339	13350	ACTGTGGTCAAA	1-10-1MOE	833
147029	14058	14069	CCCAGCCTTGTC	1-10-1MOE	848
						147029	14239	14250	CCCAGCCTTGTC	1-10-1MOE	848
147067	15560	15571	TCCTGCACTGAC	1-10-1MOE	876
						147068	15561	15572	ATCCTGCACTGA	1-10-1MOE	838
147067	15742	15753	TCCTGCACTGAC	1-10-1MOE	876
						147069	15744	15755	GATCCTGCACTG	1-10-1MOE	860
147042	16561	16572	GGTCAAAAGGGC	1-10-1MOE	866
						147042	16727	16738	GGTCAAAAGGGC	1-10-1MOE	866
147030	17619	17630	TCCCAGCCTTGT	1-10-1MOE	874
						147064	17762	17773	TGCACTGACGAG	1-10-1MOE	880
147030	17787	17798	TCCCAGCCTTGT	1-10-1MOE	874
						147064	17930	17941	TGCACTGACGAG	1-10-1MOE	880
147042	19201	19212	GGTCAAAAGGGC	1-10-1MOE	866
						147042	19369	19380	GGTCAAAAGGGC	1-10-1MOE	866
147027	21190	21201	CAGCCTTGTCGA	1-10-1MOE	843
						147028	21191	21202	CCAGCCTTGTCG	1-10-1MOE	846
147027	21358	21369	CAGCCTTGTCGA	1-10-1MOE	843
						147028	21359	21370	CCAGCCTTGTCG	1-10-1MOE	846
147070	22021	22032	TGATCCTGCACT	1-10-1MOE	878
						147070	22189	22200	TGATCCTGCACT	1-10-1MOE	878
147047	22606	22617	ACTGTGGTCAAA	1-10-1MOE	833
						147043	24318	24329	TGGTCAAAAGGG	1-10-1MOE	881
147044	24319	24330	GTGGTCAAAAGG	1-10-1MOE	869
						147045	24320	24331	TGTGGTCAAAAG	1-10-1MOE	883
147046	24321	24332	CTGTGGTCAAAA	1-10-1MOE	858
						147043	24486	24497	TGGTCAAAAGGG	1-10-1MOE	881
147044	24487	24498	GTGGTCAAAAGG	1-10-1MOE	869
						147046	24489	24500	CTGTGGTCAAAA	1-10-1MOE	858
147047	24490	24501	ACTGTGGTCAAA	1-10-1MOE	833
						147040	25065	25076	GCCGCCCAGTTC	1-10-1MOE	864
147041	25066	25077	AGCCGCCCAGTT	1-10-1MOE	834
						147046	25160	25171	CTGTGGTCAAAA	1-10-1MOE	858
147039	25232	25243	CCGCCCAGTTCC	1-10-1MOE	850

147040	25233	25244	GCCGCCCAGTTC	1-10-1MOE	864
						147041	25234	25245	AGCCGCCCAGTT	1-10-1MOE	834
147046	25328	25339	CTGTGGTCAAAA	1-10-1MOE	858
						147057	25508	25519	ATTTAATCCGAC	1-10-1MOE	839
147061	25512	25523	TGCAATTTAATC	1-10-1MOE	879
						147057	25676	25687	ATTTAATCCGAC	1-10-1MOE	839
147069	28878	28889	GATCCTGCACTG	1-10-1MOE	860
						147070	28879	28890	TGATCCTGCACT	1-10-1MOE	878
147053	30133	30144	AATCCGACTGTG	1-10-1MOE	829
						147053	30278	30289	AATCCGACTGTG	1-10-1MOE	829
147054	30864	30875	TAATCCGACTGT	1-10-1MOE	871
						147043	30985	30996	TGGTCAAAAGGG	1-10-1MOE	881
147054	31011	31022	TAATCCGACTGT	1-10-1MOE	871
						147043	31133	31144	TGGTCAAAAGGG	1-10-1MOE	881
147036	32233	32244	CCCAGTTCCCAG	1-10-1MOE	849
						147072	32372	32383	ACTGATCCTGCA	1-10-1MOE	832
147072	32520	32531	ACTGATCCTGCA	1-10-1MOE	832
						147069	33056	33067	GATCCTGCACTG	1-10-1MOE	860
147070	33057	33068	TGATCCTGCACT	1-10-1MOE	878
						147071	33058	33069	CTGATCCTGCAC	1-10-1MOE	856
147051	33126	33137	TCCGACTGTGGT	1-10-1MOE	875
						147070	33205	33216	TGATCCTGCACT	1-10-1MOE	878
147071	33206	33217	CTGATCCTGCAC	1-10-1MOE	856
						147051	33274	33285	TCCGACTGTGGT	1-10-1MOE	875
147046	33318	33329	CTGTGGTCAAAA	1-10-1MOE	858
						147049	33321	33332	CGACTGTGGTCA	1-10-1MOE	854
147051	33323	33334	TCCGACTGTGGT	1-10-1MOE	875
						147046	33466	33477	CTGTGGTCAAAA	1-10-1MOE	858
147047	33467	33478	ACTGTGGTCAAA	1-10-1MOE	833
						147051	33471	33482	TCCGACTGTGGT	1-10-1MOE	875
147046	33640	33651	CTGTGGTCAAAA	1-10-1MOE	858
						147051	33645	33656	TCCGACTGTGGT	1-10-1MOE	875
147046	33788	33799	CTGTGGTCAAAA	1-10-1MOE	858
						147051	33793	33804	TCCGACTGTGGT	1-10-1MOE	875
147059	35437	35448	CAATTTAATCCG	1-10-1MOE	840
						147060	35438	35449	GCAATTTAATCC	1-10-1MOE	861
147060	35586	35597	GCAATTTAATCC	1-10-1MOE	861
						147021	36093	36104	TGTCGATCTCCT	1-10-1MOE	882
147061	36250	36261	TGCAATTTAATC	1-10-1MOE	879
						147061	36398	36409	TGCAATTTAATC	1-10-1MOE	879
147073	37485	37496	CACTGATCCTGC	1-10-1MOE	842
						147073	37633	37644	CACTGATCCTGC	1-10-1MOE	842
147043	40214	40225	TGGTCAAAAGGG	1-10-1MOE	881
						147061	40353	40364	TGCAATTTAATC	1-10-1MOE	879
147043	40362	40373	TGGTCAAAAGGG	1-10-1MOE	881
						147061	40501	40512	TGCAATTTAATC	1-10-1MOE	879
147031	42527	42538	TTCCCAGCCTTG	1-10-1MOE	885
						147032	42528	42539	GTTCCCAGCCTT	1-10-1MOE	870
147034	42530	42541	CAGTTCCCAGCC	1-10-1MOE	844
						147031	42675	42686	TTCCCAGCCTTG	1-10-1MOE	885
147032	42676	42687	GTTCCCA GCCTT	1-10-1MOE	870

147033	42677	42688	AGTTCCCAGCCT	1-10-1MOE	836
						147034	42678	42689	CAGTTCCCAGCC	1-10-1MOE	844
147074	43848	43859	CCACTGATCCTG	1-10-1MOE	845
						147074	43996	44007	CCACTGATCCTG	1-10-1MOE	845
147051	45402	45413	TCCGACTGTGGT	1-10-1MOE	875
						147051	45550	45561	TCCGACTGTGGT	1-10-1MOE	875
147074	46125	46136	CCACTGATCCTG	1-10-1MOE	845
						147057	46313	46324	ATTTAATCCGAC	1-10-1MOE	839
147058	46314	46325	AATTTAATCCGA	1-10-1MOE	830
						147059	46315	46326	CAATTTAATCCG	1-10-1MOE	840
147061	46317	46328	TGCAATTTAATC	1-10-1MOE	879
						147057	46461	46472	ATTTAATCCGAC	1-10-1MOE	839
147059	46463	46474	CAATTTAATCCG	1-10-1MOE	840
						147061	46465	46476	TGCAATTTAATC	1-10-1MOE	879
147058	47413	47424	AATTTAATCCGA	1-10-1MOE	830
						147073	48221	48232	CACTGATCCTGC	1-10-1MOE	842
147073	48369	48380	CACTGATCCTGC	1-10-1MOE	842
						147074	48370	48381	CCACTGA TCCTG	1-10-1MOE	845
147027	48566	48577	CAGCCTTGTCGA	1-10-1MOE	843
						147027	48714	48725	CAGCCTTGTCGA	1-10-1MOE	843
147028	48715	48726	CCAGCCTTGTCG	1-10-1MOE	846
						147067	49050	49061	TCCTGCACTGAC	1-10-1MOE	876
147068	49051	49062	ATCCTGCACTGA	1-10-1MOE	838
						147067	49198	49209	TCCTGCACTGAC	1-10-1MOE	876
147073	49524	49535	CACTGATCCTGC	1-10-1MOE	842
						147073	49672	49683	CACTGATCCTGC	1-10-1MOE	842
147074	49673	49684	CCACTGATCCTG	1-10-1MOE	845
						147036	50421	50432	CCCAGTTCCCAG	1-10-1MOE	849
147036	52292	52303	CCCAGTTCCCAG	1-10-1MOE	849
						147037	52293	52304	GCCCAGTTCCCA	1-10-1MOE	863
147036	52438	52449	CCCAGTTCCCAG	1-10-1MOE	849
						147037	52439	52450	GCCCAGTTCCCA	1-10-1MOE	863
147034	53148	53159	CAGTTCCCAGCC	1-10-1MOE	844
						147034	53294	53305	CAGTTCCCAGCC	1-10-1MOE	844
147042	53445	53456	GGTCAAAAGGGC	1-10-1MOE	866
						147043	53446	53457	TGGTCAAAAGGG	1-10-1MOE	881
147044	53447	53458	GTGGTCAAAAGG	1-10-1MOE	869
						147042	53591	53602	GGTCAAAAGGGC	1-10-1MOE	866
147030	53592	53603	TCCCAGCCTTGT	1-10-1MOE	874
						147043	53592	53603	TGGTCAAAAGGG	1-10-1MOE	881
147031	53593	53604	TTCCCAGCCTTG	1-10-1MOE	885
						147044	53593	53604	GTGGTCAAAAGG	1-10-1MOE	869
147030	53738	53749	TCCCAGCCTTGT	1-10-1MOE	874
						147031	53739	53750	TTCCCAGCCTTG	1-10-1MOE	885
147040	53783	53794	GCCGCCCAGTTC	1-10-1MOE	864
						147041	53784	53795	AGCCGCCCAGTT	1-10-1MOE	834
147041	53930	53941	AGCCGCCCAGTT	1-10-1MOE	834
						147042	55008	55019	GGTCAAAAGGGC	1-10-1MOE	866
147043	55009	55020	TGGTCAAAAGGG	1-10-1MOE	881
						147042	55154	55165	GGTCAAAAGGGC	1-10-1MOE	866
147043	55155	55166	TGGTCAAAAGGG	1-10-1MOE	881

147058	55281	55292	AATTTAATCCGA	1-10-1MOE	830
						147058	55427	55438	AATTTAATCCGA	1-10-1MOE	830
147019	55682	55693	TCGATCTCCTCG	1-10-1MOE	877
						147021	55684	55695	TGTCGATCTCCT	1-10-1MOE	882
147021	55830	55841	TGTCGATCTCCT	1-10-1MOE	882
						147054	56275	56286	TAATCCGACTGT	1-10-1MOE	871
147055	56276	56287	TTAATCCGACTG	1-10-1MOE	884
						147056	56277	56288	TTTAATCCGACT	1-10-1MOE	887
147058	56279	56290	AATTTAATCCGA	1-10-1MOE	830
						147059	56280	56291	CAATTTAATCCG	1-10-1MOE	840
147060	56281	56292	GCAATTTAATCC	1-10-1MOE	861
						147061	56282	56293	TGCAATTTAATC	1-10-1MOE	879
147051	56418	56429	TCCGACTGTGGT	1-10-1MOE	875
						147053	56420	56431	AATCCGACTGTG	1-10-1MOE	829
147054	56421	56432	TAATCCGACTGT	1-10-1MOE	871
						147055	56422	56433	TTAATCCGACTG	1-10-1MOE	884
147056	56423	56434	TTTAATCCGACT	1-10-1MOE	887
						147057	56424	56435	ATTTAATCCGAC	1-10-1MOE	839
147058	56425	56436	AATTTAATCCGA	1-10-1MOE	830
						147061	56428	56439	TGCAATTTAATC	1-10-1MOE	879
147045	57118	57129	TGTGGTCAAAAG	1-10-1MOE	883
						147045	57264	57275	TGTGGTCAAAAG	1-10-1MOE	883
147046	57265	57276	CTGTGGTCAAAA	1-10-1MOE	858
						147071	58028	58039	CTGATCCTGCAC	1-10-1MOE	856
147071	58174	58185	CTGATCCTGCAC	1-10-1MOE	856
						147043	61111	61122	TGGTCAAAAGGG	1-10-1MOE	881
147071	61130	61141	CTGATCCTGCAC	1-10-1MOE	856
						147020	61226	61237	GTCGATCTCCTC	1-10-1MOE	868
147043	61257	61268	TGGTCAAAAGGG	1-10-1MOE	881
						147071	61276	61287	CTGATCCTGCAC	1-10-1MOE	856
147035	61277	61288	CCAGTTCCCAGC	1-10-1MOE	847
						147036	61278	61289	CCCAGTTCCCAG	1-10-1MOE	849
147037	61279	61290	GCCCAGTTCCCA	1-10-1MOE	863
						147038	61280	61291	CGCCCAGTTCCC	1-10-1MOE	855
147039	61281	61292	CCGCCCAGTTCC	1-10-1MOE	850
						147040	61282	61293	GCCGCCCAGTTC	1-10-1MOE	864
147071	61309	61320	CTGATCCTGCAC	1-10-1MOE	856
						147020	61372	61383	GTCGATCTCCTC	1-10-1MOE	868
147034	61422	61433	CAGTTCCCAGCC	1-10-1MOE	844
						147035	61423	61434	CCAGTTCCCAGC	1-10-1MOE	847
147036	61424	61435	CCCAGTTCCCAG	1-10-1MOE	849
						147037	61425	61436	GCCCAGTTCCCA	1-10-1MOE	863
147038	61426	61437	CGCCCAGTTCCC	1-10-1MOE	855
						147040	61428	61439	GCCGCCCAGTTC	1-10-1MOE	864
147071	61455	61466	CTGATCCTGCAC	1-10-1MOE	856
						147073	62003	62014	CACTGATCCTGC	1-10-1MOE	842
147073	62149	62160	CACTGATCCTGC	1-10-1MOE	842
						147066	63065	63076	CCTGCACTGACG	1-10-1MOE	851
147068	63067	63078	ATCCTGCACTGA	1-10-1MOE	838
						147069	63146	63157	GATCCTGCACTG	1-10-1MOE	860
147062	63207	63218	CACTGACGAGTC	1-10-1MOE	841

147066	63211	63222	CCTGCACTGACG	1-10-1MOE	851
						147057	64054	64065	ATTTAATCCGAC	1-10-1MOE	839
147036	64538	64549	CCCAGTTCCCAG	1-10-1MOE	849
						147037	64539	64550	GCCCAGTTCCCA	1-10-1MOE	863
147037	64685	64696	GCCCAGTTCCCA	1-10-1MOE	863
						147066	64864	64875	CCTGCACTGACG	1-10-1MOE	851
147067	64865	64876	TCCTGCACTGAC	1-10-1MOE	876
						147066	65010	65021	CCTGCACTGACG	1-10-1MOE	851
147067	65011	65022	TCCTGCACTGAC	1-10-1MOE	876
						147045	65017	65028	TGTGGTCAAAAG	1-10-1MOE	883
147045	65163	65174	TGTGGTCAAAAG	1-10-1MOE	883
						147046	65164	65175	CTGTGGTCAAAA	1-10-1MOE	858
147068	65408	65419	ATCCTGCACTGA	1-10-1MOE	838
						147071	65411	65422	CTGATCCTGCAC	1-10-1MOE	856
147069	65549	65560	GATCCTGCACTG	1-10-1MOE	860
						147068	65554	65565	ATCCTGCACTGA	1-10-1MOE	838
147071	65557	65568	CTGATCCTGCAC	1-10-1MOE	856
						147029	67741	67752	CCCAGCCTTGTC	1-10-1MOE	848
147030	67742	67753	TCCCAGCCTTGT	1-10-1MOE	874
						147031	67743	67754	TTCCCAGCCTTG	1-10-1MOE	885
147028	67886	67897	CCAGCCTTGTCG	1-10-1MOE	846
						147029	67887	67898	CCCAGCCTTGTC	1-10-1MOE	848
147030	67888	67899	TCCCAGCCTTGT	1-10-1MOE	874
						147031	67889	67900	TTCCCAGCCTTG	1-10-1MOE	885
147043	68867	68878	TGGTCAAAAGGG	1-10-1MOE	881
						147044	68868	68879	GTGGTCAAAAGG	1-10-1MOE	869
147045	68869	68880	TGTGGTCAAAAG	1-10-1MOE	883
						147043	69013	69024	TGGTCAAAAGGG	1-10-1MOE	881
147044	69014	69025	GTGGTCAAAAGG	1-10-1MOE	869
						147045	69015	69026	TGTGGTCAAAAG	1-10-1MOE	883
147046	69016	69027	CTGTGGTCAAAA	1-10-1MOE	858
						147071	69519	69530	CTGATCCTGCAC	1-10-1MOE	856
147072	69520	69531	ACTGATCCTGCA	1-10-1MOE	832
						147073	69521	69532	CACTGATCCTGC	1-10-1MOE	842
147071	69665	69676	CTGATCCTGCAC	1-10-1MOE	856
						147072	69666	69677	ACTGATCCTGCA	1-10-1MOE	832
147073	69667	69678	CACTGATCCTGC	1-10-1MOE	842
						147074	69668	69679	CCACTGATCCTG	1-10-1MOE	845
147066	69869	69880	CCTGCACTGACG	1-10-1MOE	851
						147066	70015	70026	CCTGCACTGACG	1-10-1MOE	851
147023	70465	70476	CTTGTCGATCTC	1-10-1MOE	859
						147023	70611	70622	CTTGTCGATCTC	1-10-1MOE	859
147062	70615	70626	CACTGACGAGTC	1-10-1MOE	841
						147063	70616	70627	GCACTGACGAGT	1-10-1MOE	862
147064	70617	70628	TGCACTGACGAG	1-10-1MOE	880
						147065	70618	70629	CTGCACTGACGA	1-10-1MOE	857
147066	70619	70630	CCTGCACTGACG	1-10-1MOE	851
						147063	70762	70773	GCACTGACGAGT	1-10-1MOE	862
147064	70763	70774	TGCACTGACGAG	1-10-1MOE	880
						147065	70764	70775	CTGCACTGACGA	1-10-1MOE	857
147066	70765	70776	CCTGCACTGACG	1-10-1MOE	851

147072	70998	71009	ACTGATCCTGCA	1-10-1MOE	832
						147073	70999	71010	CACTGATCCTGC	1-10-1MOE	842
147072	71144	71155	ACTGATCCTGCA	1-10-1MOE	832
						147073	71145	71156	CACTGATCCTGC	1-10-1MOE	842
147074	71146	71157	CCACTGATCCTG	1-10-1MOE	845
						147037	71351	71362	GCCCAGTTCCCA	1-10-1MOE	863
147038	71352	71363	CGCCCAGTTCCC	1-10-1MOE	855
						147039	71353	71364	CCGCCCAGTTCC	1-10-1MOE	850
147037	71497	71508	GCCCAGTTCCCA	1-10-1MOE	863
						147038	71498	71509	CGCCCAGTTCCC	1-10-1MOE	855
147039	71499	71510	CCGCCCAGTTCC	1-10-1MOE	850
						147061	71641	71652	TGCAATTTAATC	1-10-1MOE	879
147061	71787	71798	TGCAATTTAATC	1-10-1MOE	879

Table 17: targeting SEQ ID NO:11 and have the short antisense compounds of 1 or 2 mispairing

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						147022	177	188	TTGTCGATCTCC	1-10-1MOE	886
147023	178	189	CTTGTCGATCTC	1-10-1MOE	859
						147020	196	207	GTCGATCTCCTC	1-10-1MOE	868
147022	198	209	TTGTCGATCTCC	1-10-1MOE	886
						147024	200	211	CCTTGTCGATCT	1-10-1MOE	853
147026	202	213	AGCCTTGTCGAT	1-10-1MOE	835
						147028	204	215	CCAGCCTTGTCG	1-10-1MOE	846
147029	205	216	CCCAGCCTTGTC	1-10-1MOE	848
						147030	206	217	TCCCAGCCTTGT	1-10-1MOE	874
147036	212	223	CCCAGTTCCCAG	1-10-1MOE	849
						147073	311	322	CACTGATCCTGC	1-10-1MOE	842
147046	327	338	CTGTGGTCAAAA	1-10-1MOE	858
						147047	328	339	ACTGTGGTCAAA	1-10-1MOE	833
147048	329	340	GACTGTGGTCAA	1-10-1MOE	888
						147049	330	341	CGACTGTGGTCA	1-10-1MOE	854
147050	331	342	CCGACTGTGGTC	1-10-1MOE	889
						147051	332	343	TCCGACTGTGGT	1-10-1MOE	875
147052	333	344	ATCCGACTGTGG	1-10-1MOE	837
						147053	334	345	AATCCGACTGTG	1-10-1MOE	829
147054	335	346	TAATCCGACTGT	1-10-1MOE	871
						147055	336	347	TTAATCCGACTG	1-10-1MOE	884
147056	337	348	TTTAATCCGACT	1-10-1MOE	887
						147057	338	349	ATTTAATCCGAC	1-10-1MOE	839
147058	339	350	AATTTAATCCGA	1-10-1MOE	830
						147060	341	352	GCAATTTAATCC	1-10-1MOE	861
147061	342	353	TGCAATTTAATC	1-10-1MOE	879
						147062	1024	1035	CACTGACGAGTC	1-10-1MOE	841
147063	1025	1036	GCACTGACGAGT	1-10-1MOE	862
						147068	1030	1041	ATCCTGCACTGA	1-10-1MOE	838
147071	1033	1044	CTGATCCTGCAC	1-10-1MOE	856

147073	1035	1046	CACTGATCCTGC	1-10-1MOE	842
						147074	1036	1047	CCACTGATCCTG	1-10-1MOE	845
147067	1091	1102	TCCTGCACTGAC	1-10-1MOE	876
						147024	1891	1902	CCTTGTCGATCT	1-10-1MOE	853
147026	1893	1904	AGCCTTGTCGAT	1-10-1MOE	835
						147029	1896	1907	CCCAGCCTTGTC	1-10-1MOE	848
147036	1903	1914	CCCAGTTCCCAG	1-10-1MOE	849
						147039	1906	1917	CCGCCCAGTTCC	1-10-1MOE	850
147019	1994	2005	TCGATCTCCTCG	1-10-1MOE	877
						401385	2815	2828	CCCAGTGGGTTTGA	2-10-2MOE	890
147033	5265	5276	AGTTCCCAGCCT	1-10-1MOE	836
						147033	5373	5384	AGTTCCCAGCCT	1-10-1MOE	836
147060	7168	7179	GCAATTTAATCC	1-10-1MOE	861
						147053	10527	10538	AATCCGACTGTG	1-10-1MOE	829
147053	10635	10646	AATCCGACTGTG	1-10-1MOE	829
						147070	11604	11615	TGATCCTGCACT	1-10-1MOE	878
147071	11612	11623	CTGATCCTGCAC	1-10-1MOE	856
						147072	12294	12305	ACTGATCCTGCA	1-10-1MOE	832
147072	12299	12310	ACTGATCCTGCA	1-10-1MOE	832
						147052	12938	12949	ATCCGACTGTGG	1-10-1MOE	837
147052	13119	13130	ATCCGACTGTGG	1-10-1MOE	837
						147047	13158	13169	ACTGTGGTCAAA	1-10-1MOE	833
147048	13159	13170	GACTGTGGTCAA	1-10-1MOE	888
						147049	13160	13171	CGACTGTGGTCA	1-10-1MOE	854
147048	13340	13351	GACTGTGGTCAA	1-10-1MOE	888
						147049	13341	13352	CGACTGTGGTCA	1-10-1MOE	854
147051	13343	13354	TCCGACTGTGGT	1-10-1MOE	875
						147061	13497	13508	TGCAATTTAATC	1-10-1MOE	879
147069	15562	15573	GATCCTGCACTG	1-10-1MOE	860
						147068	15743	15754	ATCCTGCACTGA	1-10-1MOE	838
147049	17181	17192	CGACTGTGGTCA	1-10-1MOE	854
						147049	17349	17360	CGACTGTGGTCA	1-10-1MOE	854
147047	22438	22449	ACTGTGGTCAAA	1-10-1MOE	833
						147047	24322	24333	ACTGTGGTCAAA	1-10-1MOE	833
147045	24488	24499	TGTGGTCAAAAG	1-10-1MOE	883
						147039	25064	25075	CCGCCCAGTTCC	1-10-1MOE	850
147057	25508	25519	ATTTAATCCGAC	1-10-1MOE	839
						147057	25676	25687	ATTTAATCCGAC	1-10-1MOE	839
147061	25680	25691	TGCAATTTAATC	1-10-1MOE	879
						147069	28731	28742	GATCCTGCACTG	1-10-1MOE	860
147052	30132	30143	ATCCGACTGTGG	1-10-1MOE	837
						147052	30277	30288	ATCCGACTGTGG	1-10-1MOE	837
147036	32085	32096	CCCAGTTCCCAG	1-10-1MOE	849
						147072	32520	32531	ACTGATCCTGCA	1-10-1MOE	832
147071	33058	33069	CTGATCCTGCAC	1-10-1MOE	856
						147050	33125	33136	CCGACTGTGGTC	1-10-1MOE	889
147069	33204	33215	GATCCTGCACTG	1-10-1MOE	860
						147050	33273	33284	CCGACTGTGGTC	1-10-1MOE	889
147047	33319	33330	ACTGTGGTCAAA	1-10-1MOE	833
						147050	33322	33333	CCGACTGTGGTC	1-10-1MOE	889
147052	33324	33335	ATCCGACTGTGG	1-10-1MOE	837

147049	33469	33480	CGACTGTGGTCA	1-10-1MOE	854
						147050	33470	33481	CCGACTGTGGTC	1-10-1MOE	889
147052	33472	33483	ATCCGACTGTGG	1-10-1MOE	837
						147047	33641	33652	ACTGTGGTCAAA	1-10-1MOE	833
147047	33789	33800	ACTGTGGTCAAA	1-10-1MOE	833
						147059	35585	35596	CAATTTAATCCG	1-10-1MOE	840
147021	36241	36252	TGTCGATCTCCT	1-10-1MOE	882
						147073	37633	37644	CACTGATCCTGC	1-10-1MOE	842
147033	42529	42540	AGTTCCCAGCCT	1-10-1MOE	836
						147050	45401	45412	CCGACTGTGGTC	1-10-1MOE	889
147050	45549	45560	CCGACTGTGGTC	1-10-1MOE	889
						147074	46125	46136	CCACTGATCCTG	1-10-1MOE	845
147057	46313	46324	ATTTAATCCGAC	1-10-1MOE	839
						147058	46462	46473	AATTTAATCCGA	1-10-1MOE	830
147058	47413	47424	AATTTAATCCGA	1-10-1MOE	830
						147058	47561	47572	AATTTAATCCGA	1-10-1MOE	830
147073	48221	48232	CACTGATCCTGC	1-10-1MOE	842
						147073	48369	48380	CACTGATCCTGC	1-10-1MOE	842
147028	48567	48578	CCAGCCTTGTCG	1-10-1MOE	846
						147068	49199	49210	ATCCTGCACTGA	1-10-1MOE	838
147036	50273	50284	CCCAGTTCCCAG	1-10-1MOE	849
						147040	53929	53940	GCCGCCCAGTTC	1-10-1MOE	864
147047	54769	54780	ACTGTGGTCAAA	1-10-1MOE	833
						147048	54770	54781	GACTGTGGTCAA	1-10-1MOE	888
147047	54915	54926	ACTGTGGTCAAA	1-10-1MOE	833
						147048	54916	54927	GACTGTGGTCAA	1-10-1MOE	888
147019	55828	55839	TCGATCTCCTCG	1-10-1MOE	877
						147047	56268	56279	ACTGTGGTCAAA	1-10-1MOE	833
147048	56269	56280	GACTGTGGTCAA	1-10-1MOE	888
						147049	56270	56281	CGACTGTGGTCA	1-10-1MOE	854
147050	56271	56282	CCGACTGTGGTC	1-10-1MOE	889
						147051	56272	56283	TCCGACTGTGGT	1-10-1MOE	875
147052	56273	56284	ATCCGACTGTGG	1-10-1MOE	837
						147053	56274	56285	AATCCGACTGTG	1-10-1MOE	829
147056	56277	56288	TTTAATCCGACT	1-10-1MOE	887
						147057	56278	56289	ATTTAATCCGAC	1-10-1MOE	839
147047	56414	56425	ACTGTGGTCAAA	1-10-1MOE	833
						147048	56415	56426	GACTGTGGTCAA	1-10-1MOE	888
147049	56416	56427	CGACTGTGGTCA	1-10-1MOE	854
						147050	56417	56428	CCGACTGTGGTC	1-10-1MOE	889
147052	56419	56430	ATCCGACTGTGG	1-10-1MOE	837
						147057	56424	56435	ATTTAATCCGAC	1-10-1MOE	839
147058	56425	56436	AATTTAATCCGA	1-10-1MOE	830
						147059	56426	56437	CAATTTAATCCG	1-10-1MOE	840
147060	56427	56438	GCAATTTAATCC	1-10-1MOE	861
						147046	57119	57130	CTGTGGTCAAAA	1-10-1MOE	858
147071	58174	58185	CTGATCCTGCAC	1-10-1MOE	856
						147071	61130	61141	CTGATCCTGCAC	1-10-1MOE	856
147034	61276	61287	CAGTTCCCAGCC	1-10-1MOE	844
						147071	61309	61320	CTGATCCTGCAC	1-10-1MOE	856
147039	61427	61438	CCGCCCAGTTCC	1-10-1MOE	850

147071	61455	61466	CTGATCCTGCAC	1-10-1MOE	856
						147073	62003	62014	CACTGATCCTGC	1-10-1MOE	842
147062	63061	63072	CACTGACGAGTC	1-10-1MOE	841
						147068	63213	63224	ATCCTGCACTGA	1-10-1MOE	838
147069	63292	63303	GATCCTGCACTG	1-10-1MOE	860
						147057	64054	64065	ATTTAATCCGAC	1-10-1MOE	839
147057	64200	64211	ATTTAATCCGAC	1-10-1MOE	839
						147070	64427	64438	TGATCCTGCACT	1-10-1MOE	878
147070	64573	64584	TGATCCTGCACT	1-10-1MOE	878
						147036	64684	64695	CCCAGTTCCCAG	1-10-1MOE	849
147046	65018	65029	CTGTGGTCAAAA	1-10-1MOE	858
						147071	65557	65568	CTGATCCTGCAC	1-10-1MOE	856
147069	65695	65706	GATCCTGCACTG	1-10-1MOE	860
						147047	66163	66174	ACTGTGGTCAAA	1-10-1MOE	833
147047	66309	66320	ACTGTGGTCAAA	1-10-1MOE	833
						147028	67740	67751	CCAGCCTTGTCG	1-10-1MOE	846
147046	68870	68881	CTGTGGTCAAAA	1-10-1MOE	858
						147047	68871	68882	ACTGTGGTCAAA	1-10-1MOE	833
147048	68872	68883	GACTGTGGTCAA	1-10-1MOE	888
						147049	68873	68884	CGACTGTGGTCA	1-10-1MOE	854
147047	69017	69028	ACTGTGGTCAAA	1-10-1MOE	833
						147048	69018	69029	GACTGTGGTCAA	1-10-1MOE	888
147049	69019	69030	CGACTGTGGTCA	1-10-1MOE	854
						147071	69519	69530	CTGATCCTGCAC	1-10-1MOE	856
147073	69521	69532	CACTGATCCTGC	1-10-1MOE	842
						147071	69665	69676	CTGATCCTGCAC	1-10-1MOE	856
147072	69666	69677	ACTGATCCTGCA	1-10-1MOE	832
						147024	70466	70477	CCTTGTCGATCT	1-10-1MOE	853
147024	70612	70623	CCTTGTCGATCT	1-10-1MOE	853
						147062	70761	70772	CACTGACGAGTC	1-10-1MOE	841
147072	70998	71009	ACTGATCCTGCA	1-10-1MOE	832
						147073	70999	71010	CACTGATCCTGC	1-10-1MOE	842
147072	71144	71155	ACTGATCCTGCA	1-10-1MOE	832
						147073	71145	71156	CACTGATCCTGC	1-10-1MOE	842
147048	71366	71377	GACTGTGGTCAA	1-10-1MOE	888
						147048	71512	71523	GACTGTGGTCAA	1-10-1MOE	888

Table 18: the short antisense compounds of targeting SEQ ID NO:12

ISIS NO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	Seq ID NO
						398163	20	31	ATGTCAACCGGC	1-10-1MOE	908
384545	23	34	CAAGTAGGATGT	1-10-1MOE	951
						147705	159	170	CGGTTTTTGTTC	1-10-1MOE	1002
147703	245	256	TGGCTTCATGTC	1-10-1MOE	971
						398090	283	296	TTGTTCTTAGGAAG	2-10-2MOE	972
147704	285	296	TTGTTCTTAGGA	1-10-1MOE	1012
						147705	291	302	CGGTTTTTGTTC	1-10-1MOE	1002
147709	311	322	CCATTTTTATCA	1-10-1MOE	978

147733	349	360	TTCTTGATGTCC	1-10-1MOE	891
						147707	360	371	TAGTCATTATCT	1-10-1MOE	977
147708	366	377	TTGATATAGTCA	1-10-1MOE	997
						390030	381	392	TTTATAAAACTG	1-10-1MOE	1074
147709	386	397	CCATTTTTATCA	1-10-1MOE	978
						147081	393	404	GCTCCTTCCACT	1-10-1MOE	1006
398091	393	406	GGGCTTCTTCCATT	2-10-2MOE	979
						398166	395	406	GGGCTTCTTCCA	1-10-1MOE	1070
147709	418	429	CCATTTTTATCA	1-10-1MOE	978
						147711	425	436	AAGGGCCCTGGG	1-10-1MOE	1040
147712	461	472	ACACCATCTCCC	1-10-1MOE	1005
						147713	466	477	CTCCCACACCAT	1-10-1MOE	985
147714	471	482	TTCTGCTCCCAC	1-10-1MOE	986
						147715	496	507	GTTGAGCATGAC	1-10-1MOE	1077
147716	521	532	TTAACGAGCCTT	1-10-1MOE	949
						147717	574	585	ATCTTCAGAGAT	1-10-1MOE	996
147717	607	618	ATCTTCAGAGAT	1-10-1MOE	996
						147708	612	623	TTGATATAGTCA	1-10-1MOE	997
147718	621	632	TAATATGACTTG	1-10-1MOE	998
						147746	625	636	TAAAAACAACAA	1-10-1MOE	1073
398167	704	715	CAGGCCATGTGG	1-10-1MOE	1059
						398092	705	718	AGTCAGGCCATGTG	2-10-2MOE	1060
147723	715	726	GACTCCAAAGTC	1-10-1MOE	892
						398093	758	771	TCGGACTTTGAAAA	2-10-2MOE	1009
398168	760	771	TCGGACTTTGAA	1-10-1MOE	1008
						147738	780	791	TGGGTGGCCGGG	1-10-1MOE	1069
398094	848	861	ATCAGCCAGACAGA	2-10-2MOE	1010
						398169	849	860	TCAGCCAGACAG	1-10-1MOE	909
398164	873	884	TTGTCGATCTGC	1-10-1MOE	1014
						147735	973	984	GGAGAAGCGCAG	1-10-1MOE	1016
147737	984	995	ACAGCCAGGTAG	1-10-1MOE	1067
						368369	1025	1040	TCCTGCACTGACGAGT	3-10-3MOE	893
368372	1031	1046	CACTGATCCTGCACTG	3-10-3MOE	894
						368353	1033	1046	CACTGATCCTGCAC	2-10-2MOE	1007
368354	1035	1048	TCCACTGATCCTGC	2-10-2MOE	1024
						368388	1035	1050	CTTCCACTGATCCTTA	3-10-3MOE	895
368355	1036	1049	TTCCACTGATCCTG	2-10-2MOE	1025
						368356	1037	1050	CTTCCACTGATCCT	2-10-2MOE	1027
368376	1037	1052	TCCTTCCACTGATCCT	3-10-3MOE	1028
						147076	1038	1049	TTCCACTGATCC	1-10-1MOE	1029
368357	1038	1051	CCTTCCACTGATCC	2-10-2MOE	1046
						147077	1039	1050	CTTCCACTGATC	1-10-1MOE	1047
368358	1039	1052	TCCTTCCACTGATC	2-10-2MOE	1031
						368378	1039	1054	GCTCCTTCCACTGATC	3-10-3MOE	1032
368359	1041	1054	GCTCCTTCCACTGA	2-10-2MOE	1033
						147080	1042	1053	CTCCTTCCACTG	1-10-1MOE	1021
147081	1043	1054	GCTCCTTCCACT	1-10-1MOE	1006
						368360	1043	1056	AAGCTCCTTCCACT	2-10-2MOE	1035
368380	1043	1058	GAAAGCTCCTTCCACT	3-10-3MOE	896
						147082	1044	1055	AGCTCCTTCCAC	1-10-1MOE	1036
368381	1045	1060	GGGAAAGCTCCTTCCA	3-10-3MOE	1037
						147739	1107	1118	CGTTTGGGTGGC	1-10-1MOE	1023
147741	1165	1176	CACCCACTGGTG	1-10-1MOE	1055
						398097	1194	1207	GGCAGTCTTTATCC	2-10-2MOE	897

147742	1273	1284	AACTTCAGTGTC	1-10-1MOE	1041
						147743	1388	1399	AGGGCTTCCAGT	1-10-1MOE	1042
147744	1392	1403	AGGAAGGGCTTC	1-10-1MOE	1043
						147745	1398	1409	TTGACCAGGAAG	1-10-1MOE	1058
398157	1455	1468	GGAAACATACCCTG	2-10-2MOE	1045
						398167	1475	1486	CAGGCCATGTGG	1-10-1MOE	1059
398092	1476	1489	AGTCAGGCCATGTG	2-10-2MOE	1060
						368357	1596	1609	CCTTCCACTGATCC	2-10-2MOE	1046
398160	1691	1704	GAATAGGTTAAGGC	2-10-2MOE	1048
						398163	1711	1722	ATGTCAACCGGC	1-10-1MOE	908
147746	1750	1761	TAAAAACAACAA	1-10-1MOE	1073
						389949	1777	1788	GCGCGAGCCCGA	1-10-1MOE	1061
398161	1790	1803	AACAATGTGTTGTA	2-10-2MOE	1049
						147746	1799	1810	TAAAAACAACAA	1-10-1MOE	1073
398163	1819	1830	ATGTCAACCGGC	1-10-1MOE	908
						389950	1848	1859	CCCTGAAGGTTC	1-10-1MOE	1063
398164	1889	1900	TTGTCGATCTGC	1-10-1MOE	1014
						147702	1917	1928	CTGGTAAATAGC	1-10-1MOE	898
147088	1971	1982	CCCTCTACACCA	1-10-1MOE	1050
						398102	2003	2016	CTACCTGAGGATTT	2-10-2MOE	899
398103	2010	2023	CCCAGTACTACCTG	2-10-2MOE	900
						147737	2386	2397	ACAGCCAGGTAG	1-10-1MOE	1067
398095	2407	2420	CATCAGCAAGAGGC	2-10-2MOE	1011
						398106	2441	2454	TGGAAAACTGCACC	2-10-2MOE	1068
147745	2497	2508	TTGACCAGGAAG	1-10-1MOE	1058
						147712	2499	2510	ACACCATCTCCC	1-10-1MOE	1005
147712	2607	2618	ACACCATCTCCC	1-10-1MOE	1005
						147745	2689	2700	TTGACCAGGAAG	1-10-1MOE	1058
398167	2706	2717	CAGGCCATGTGG	1-10-1MOE	1059
						398092	2707	2720	AGTCAGGCCATGTG	2-10-2MOE	1060
398166	2966	2977	GGGCTTCTTCCA	1-10-1MOE	1070
						147091	2992	3003	GTTCCCTCTACA	1-10-1MOE	1004
147092	2993	3004	TGTTCCCTCTAC	1-10-1MOE	901
						389949	3008	3019	GCGCGAGCCCGA	1-10-1MOE	1061
147087	3149	3160	CCTCTACACCAG	1-10-1MOE	982
						147088	3150	3161	CCCTCTACACCA	1-10-1MOE	1050
398113	3160	3173	AGGAGGTTAAACCA	2-10-2MOE	905
						147087	3257	3268	CCTCTACACCAG	1-10-1MOE	982
147088	3258	3269	CCCTCTACACCA	1-10-1MOE	1050
						147737	3591	3602	ACAGCCAGGTAG	1-10-1MOE	1067
147737	3617	3628	ACAGCCAGGTAG	1-10-1MOE	1067
						147079	3637	3648	TCCTTCCACTGA	1-10-1MOE	1001
147080	3638	3649	CTCCTTCCACTG	1-10-1MOE	1021
						398095	3638	3651	CATCAGCAAGAGGC	2-10-2MOE	1011
398106	3672	3685	TGGAAAACTGCACC	2-10-2MOE	1068
						398107	3678	3691	TATTCCTGGAAAAC	2-10-2MOE	902
147691	3806	3817	GAGGTGGGAAAA	1-10-1MOE	966
						147683	3848	3859	GCTTACGATTGT	1-10-1MOE	922
147738	3853	3864	TGGGTGGCCGGG	1-10-1MOE	1069
						398167	3926	3937	CAGGCCATGTGG	1-10-1MOE	1059
398109	3945	3958	CAAGAAGTGTGGTT	2-10-2MOE	903
						398167	4034	4045	CAGGCCATGTGG	1-10-1MOE	1059
398110	4083	4096	GTTCCCTTTGCAGG	2-10-2MOE	952
						398111	4168	4181	GTGAAAATGCTGGC	2-10-2MOE	904

147706	4238	4249	GCTGACATCTCG	1-10-1MOE	1071
						398112	4282	4295	CAGCCTGGCACCTA	2-10-2MOE	1072
147746	4315	4326	TAAAAACAACAA	1-10-1MOE	1073
						398113	4391	4404	AGGAGGTTAAACCA	2-10-2MOE	905
398115	4484	4497	AGTAAATATTGGCT	2-10-2MOE	1076
						390030	4491	4502	TTTATAAAACTG	1-10-1MOE	1074
390030	4537	4548	TTTATAAAACTG	1-10-1MOE	1074
						147703	5034	5045	TGGCTTCATGTC	1-10-1MOE	971
147684	5035	5046	ACCCAGTCAGGG	1-10-1MOE	964
						398125	5075	5088	CAGTAAGGAATTTT	2-10-2MOE	913
147696	5083	5094	TGGATGATTGGC	1-10-1MOE	906
						147684	5143	5154	ACCCAGTCAGGG	1-10-1MOE	964
147712	5366	5377	ACACCATCTCCC	1-10-1MOE	1005
						147714	5416	5427	TTCTGCTCCCAC	1-10-1MOE	986
398128	5443	5456	CTAAATTTAGTTCA	2-10-2MOE	911
						147712	5474	5485	ACACCATCTCCC	1-10-1MOE	1005
147746	5498	5509	TAAAAACAACAA	1-10-1MOE	1073
						147714	5524	5535	TTCTGCTCCCAC	1-10-1MOE	986
147736	5600	5611	AGGTAGGAGAAG	1-10-1MOE	963
						147085	5762	5773	TCTACACCAGGT	1-10-1MOE	961
147679	5825	5836	CAAAAGGATCCC	1-10-1MOE	907
						390030	6803	6814	TTTATAAAACTG	1-10-1MOE	1074
398142	6885	6898	CCAGCACACTGGAA	2-10-2MOE	923
						398142	6994	7007	CCAGCACACTGGAA	2-10-2MOE	923
398166	7306	7317	GGGCTTCTTCCA	1-10-1MOE	1070
						147684	7551	7562	ACCCAGTCAGGG	1-10-1MOE	964
147085	8308	8319	TCTACACCAGGT	1-10-1MOE	961
						147085	8416	8427	TCTACACCAGGT	1-10-1MOE	961
398163	8473	8484	ATGTCAACCGGC	1-10-1MOE	908
						147718	8523	8534	TAATATGACTTG	1-10-1MOE	998
147718	8631	8642	TAATATGACTTG	1-10-1MOE	998
						147691	8806	8817	GAGGTGGGAAAA	1-10-1MOE	966
147728	8835	8846	GCCAGACAGAAG	1-10-1MOE	1013
						147728	8943	8954	GCCAGACAGAAG	1-10-1MOE	1013
398169	8946	8957	TCAGCCAGACAG	1-10-1MOE	909
						147742	9060	9071	AACTTCAGTGTC	1-10-1MOE	1041
404136	9162	9175	TAAGTGTCCCTTTG	2-10-2MOE	910
						147746	9963	9974	TAAAAACAACAA	1-10-1MOE	1073
147746	9966	9977	TAAAAACAACAA	1-10-1MOE	1073
						147746	9969	9980	TAAAAACAACAA	1-10-1MOE	1073
147746	9991	10002	TAAAAACAACAA	1-10-1MOE	1073
						147746	10071	10082	TAAAAACAACAA	1-10-1MOE	1073
147746	10074	10085	TAAAAACAACAA	1-10-1MOE	1073
						147746	10077	10088	TAAAAACAACAA	1-10-1MOE	1073
390030	10170	10181	TTTATAAAACTG	1-10-1MOE	1074
						147084	10220	10231	CTACACCAGGTC	1-10-1MOE	993
390030	10278	10289	TTTATAAAACTG	1-10-1MOE	1074
						147085	10329	10340	TCTACACCAGGT	1-10-1MOE	961
147711	10684	10695	AAGGGCCCTGGG	1-10-1MOE	1040
						147711	10792	10803	AAGGGCCCTGGG	1-10-1MOE	1040
398128	11333	11346	CTAAATTTAGTTCA	2-10-2MOE	911
						147707	11960	11971	TAGTCATTATCT	1-10-1MOE	977
147707	11965	11976	TAGTCATTATCT	1-10-1MOE	977
						147090	12013	12024	TTCCCTCTACAC	1-10-1MOE	955

398096	12146	12159	GGAGAAGCGCAGCT	2-10-2MOE	1015
						398166	12214	12225	GGGCTTCTTCCA	1-10-1MOE	1070
398135	12308	12321	GACTACATTTTACA	2-10-2MOE	912
						147741	12389	12400	CACCCACTGGTG	1-10-1MOE	1055
398125	12431	12444	CAGTAAGGAATTTT	2-10-2MOE	913
						147714	12585	12596	TTCTGCTCCCAC	1-10-1MOE	986
147718	12594	12605	TAATATGACTTG	1-10-1MOE	998
						398125	12612	12625	CAGTAAGGAATTTT	2-10-2MOE	913
147737	12803	12814	ACAGCCAGGTAG	1-10-1MOE	1067
						147746	12876	12887	TAAAAACAACAA	1-10-1MOE	1073
147691	12900	12911	GAGGTGGGAAAA	1-10-1MOE	966
						398137	13111	13124	TGTGTCCCTCAGTC	2-10-2MOE	914
398138	13254	13267	AACATCAAGCTTGA	2-10-2MOE	931
						398137	13292	13305	TGTGTCCCTCAGTC	2-10-2MOE	914
398138	13435	13448	AACATCAAGCTTGA	2-10-2MOE	931
						389764	14020	14031	CTGCAACATGAT	1-9-2MOE	1018
389948	14067	14078	CCGTTGGACCCC	1-10-1MOE	915
						389948	14248	14259	CCGTTGGACCCC	1-10-1MOE	915
147738	14279	14290	TGGGTGGCCGGG	1-10-1MOE	1069
						147698	14572	14583	CCCGCCACCACC	1-10-1MOE	928
147717	14750	14761	ATCTTCAGAGAT	1-10-1MOE	996
						147717	14932	14943	ATCTTCAGAGAT	1-10-1MOE	996
398167	15374	15385	CAGGCCATGTGG	1-10-1MOE	1059
						147736	16444	16455	AGGTAGGAGAAG	1-10-1MOE	963
147746	16510	16521	TAAAAACAACAA	1-10-1MOE	1073
						147738	16590	16601	TGGGTGGCCGGG	1-10-1MOE	1069
147746	16676	16687	TAAAAACAACAA	1-10-1MOE	1073
						398167	16797	16808	CAGGCCATGTGG	1-10-1MOE	1059
398144	16911	16924	GACAGCTTCTATAA	2-10-2MOE	916
						389764	17096	17107	CTGCAACATGAT	1-9-2MOE	1018
147709	17238	17249	CCATTTTTATCA	1-10-1MOE	978
						147709	17406	17417	CCATTTTTATCA	1-10-1MOE	978
147695	17466	17477	TCATTCCCCACT	1-10-1MOE	984
						147746	17497	17508	TAAAAACAACAA	1-10-1MOE	1073
147088	17539	17550	CCCTCTACACCA	1-10-1MOE	1050
						147711	17808	17819	AAGGGCCCTGGG	1-10-1MOE	1040
147711	17976	17987	AAGGGCCCTGGG	1-10-1MOE	1040
						398139	18049	18062	AGTGACTGACCACA	2-10-2MOE	917
398139	18217	18230	AGTGACTGACCACA	2-10-2MOE	917
						398140	18596	18609	GTAGCATAGAGCCT	2-10-2MOE	918
398140	18764	18777	GTAGCATAGAGCCT	2-10-2MOE	918
						398167	18927	18938	CAGGCCATGTGG	1-10-1MOE	1059
398141	18947	18960	CAGATCTTGTCAAG	2-10-2MOE	919
						398167	19095	19106	CAGGCCATGTGG	1-10-1MOE	1059
398141	19115	19128	CAGATCTTGTCAAG	2-10-2MOE	919
						147746	19207	19218	TAAAAACAACAA	1-10-1MOE	1073
147711	19508	19519	AAGGGCCCTGGG	1-10-1MOE	1040
						147729	19554	19565	GTAAGAGGCAGG	1-10-1MOE	920
147718	19617	19628	TAATATGACTTG	1-10-1MOE	998
						390030	19618	19629	TTTATAAAACTG	1-10-1MOE	1074
147701	19671	19682	CCATGGCGGGAC	1-10-1MOE	921
						147711	19676	19687	AAGGGCCCTGGG	1-10-1MOE	1040
147718	19785	19796	TAATATGACTTG	1-10-1MOE	998
						147079	20515	20526	TCCTTCCACTGA	1-10-1MOE	1001

389764	20620	20631	CTGCAACATGAT	1-9-2MOE	1018
						398142	20653	20666	CCAGCACACTGGAA	2-10-2MOE	923
147078	20682	20693	CCTTCCACTGAT	1-10-1MOE	1044
						147079	20683	20694	TCCTTCCACTGA	1-10-1MOE	1001
147080	20704	20715	CTCCTTCCACTG	1-10-1MOE	1021
						147081	20705	20716	GCTCCTTCCACT	1-10-1MOE	1006
389965	20788	20799	CTGCAACATGAT	1-10-1MOE	1018
						147746	20870	20881	TAAAAACAACAA	1-10-1MOE	1073
147746	21038	21049	TAAAAACAACAA	1-10-1MOE	1073
						147717	21080	21091	ATCTTCAGAGAT	1-10-1MOE	996
147076	21222	21233	TTCCACTGATCC	1-10-1MOE	1029
						398094	21441	21454	ATCAGCCAGACAGA	2-10-2MOE	1010
147746	21633	21644	TAAAAACAACAA	1-10-1MOE	1073
						147738	21884	21895	TGGGTGGCCGGG	1-10-1MOE	1069
147683	21939	21950	GCTTACGATTGT	1-10-1MOE	922
						147743	22213	22224	AGGGCTTCCAGT	1-10-1MOE	1042
147736	22759	22770	AGGTAGGAGAAG	1-10-1MOE	963
						147736	22927	22938	AGGTAGGAGAAG	1-10-1MOE	963
398142	23008	23021	CCAGCACACTGGAA	2-10-2MOE	923
						398147	23784	23797	CTACAGGACAATAC	2-10-2MOE	957
398147	23952	23965	CTACAGGACAATAC	2-10-2MOE	957
						147713	24434	24445	CTCCCACACCAT	1-10-1MOE	985
389965	24543	24554	CTGCAACATGAT	1-10-1MOE	1018
						147713	24602	24613	CTCCCACACCAT	1-10-1MOE	985
389965	24711	24722	CTGCAACATGAT	1-10-1MOE	1018
						147746	25384	25395	TAAAAACAACAA	1-10-1MOE	1073
398143	25505	25518	GTCAGTCCCAGCTA	2-10-2MOE	924
						147691	25610	25621	GAGGTGGGAAAA	1-10-1MOE	966
398130	25672	25685	TTAGTATGACAGCT	2-10-2MOE	925
						147746	25810	25821	TAAAAACAACAA	1-10-1MOE	1073
147746	25978	25989	TAAAAACAACAA	1-10-1MOE	1073
						147746	26172	26183	TAAAAACAACAA	1-10-1MOE	1073
398151	26718	26731	TCAGTGTAGGAAGA	2-10-2MOE	926
						147728	26917	26928	GCCAGACAGAAG	1-10-1MOE	1013
398152	27708	27721	TGAATATACAGATG	2-10-2MOE	927
						147698	28629	28640	CCCGCCACCACC	1-10-1MOE	928
389965	28714	28725	CTGCAACATGAT	1-10-1MOE	1018
						389764	28714	28725	CTGCAACATGAT	1-9-2MOE	1018
389764	28861	28872	CTGCAACATGAT	1-9-2MOE	1018
						390030	29945	29956	TTTATAAAACTG	1-10-1MOE	1074
147744	30654	30665	AGGAAGGGCTTC	1-10-1MOE	1043
						147093	30836	30847	TTGTTCCCTCTA	1-10-1MOE	929
147746	30957	30968	TAAAAACAACAA	1-10-1MOE	1073
						147746	31105	31116	TAAAAACAACAA	1-10-1MOE	1073
390030	31477	31488	TTTATAAAACTG	1-10-1MOE	1074
						384545	31829	31840	CAAGTAGGATGT	1-10-1MOE	951
384545	31977	31988	CAAGTAGGATGT	1-10-1MOE	951
						401382	32094	32107	TCTACCTGAGTCCA	2-10-2MOE	930
147089	32387	32398	TCCCTCTACACC	1-10-1MOE	956
						389950	32949	32960	CCCTGAAGGTTC	1-10-1MOE	1063
398165	33002	33013	GTTCTTAGGAAG	1-10-1MOE	968
						147081	33073	33084	GCTCCTTCCACT	1-10-1MOE	1006
147082	33074	33085	AGCTCCTTCCAC	1-10-1MOE	1036
						389950	33097	33108	CCCTGAAGGTTC	1-10-1MOE	1063

147736	33160	33171	AGGTAGGAGAAG	1-10-1MOE	963
						147081	33221	33232	GCTCCTTCCACT	1-10-1MOE	1006
368360	33221	33234	AAGCTCCTTCCACT	2-10-2MOE	1035
						147082	33222	33233	AGCTCCTTCCAC	1-10-1MOE	1036
398138	33244	33257	AACATCAAGCTTGA	2-10-2MOE	931
						147746	33250	33261	TAAAAACAACAA	1-10-1MOE	1073
398138	33392	33405	AACATCAAGCTTGA	2-10-2MOE	931
						401383	33588	33601	GATCACCTTCAGAG	2-10-2MOE	932
147746	33886	33897	TAAAAACAACAA	1-10-1MOE	1073
						147746	34606	34617	TAAAAACAACAA	1-10-1MOE	1073
398165	34704	34715	GTTCTTAGGAAG	1-10-1MOE	968
						147717	34745	34756	ATCTTCAGAGAT	1-10-1MOE	996
147746	34754	34765	TAAAAACAACAA	1-10-1MOE	1073
						398165	34852	34863	GTTCTTAGGAAG	1-10-1MOE	968
147717	34893	34904	ATCTTCAGAGAT	1-10-1MOE	996
						401384	34905	34918	TGAACACATCACTA	2-10-2MOE	933
147738	35391	35402	TGGGTGGCCGGG	1-10-1MOE	1069
						147736	35396	35407	AGGTAGGAGAAG	1-10-1MOE	963
147738	35539	35550	TGGGTGGCCGGG	1-10-1MOE	1069
						147691	35554	35565	GAGGTGGGAAAA	1-10-1MOE	966
147691	35702	35713	GAGGTGGGAAAA	1-10-1MOE	966
						147746	35814	35825	TAAAAACAACAA	1-10-1MOE	1073
401385	36109	36122	CCCAGTGGGTTTGA	2-10-2MOE	890
						147691	36360	36371	GAGGTGGGAAAA	1-10-1MOE	966
147746	36416	36427	TAAAAACAACAA	1-10-1MOE	1073
						147731	36620	36631	TTTCCTCTTGTC	1-10-1MOE	934
147714	37881	37892	TTCTGCTCCCAC	1-10-1MOE	986
						147714	38029	38040	TTCTGCTCCCAC	1-10-1MOE	986
147681	38512	38523	ATGTCATTAAAC	1-10-1MOE	965
						401386	38516	38529	TAATTGATGTCAAT	2-10-2MOE	935
401387	38518	38531	AGTAATTGATGTCA	2-10-2MOE	936
						401388	38520	38533	ACAGTAATTGATGT	2-10-2MOE	937
401389	38522	38535	TTACAGTAATTGAT	2-10-2MOE	938
						401390	38524	38537	ACTTACAGTAATTG	2-10-2MOE	939
401391	38526	38539	AGACTTACAGTAAT	2-10-2MOE	940
						401392	38528	38541	TCAGACTTACAGTA	2-10-2MOE	941
401393	38530	38543	AATCAGACTTACAG	2-10-2MOE	942
						401394	38532	38545	TGAATCAGACTTAC	2-10-2MOE	943
401395	38534	38547	AATGAATCAGACTT	2-10-2MOE	944
						147738	38909	38920	TGGGTGGCCGGG	1-10-1MOE	1069
147738	39057	39068	TGGGTGGCCGGG	1-10-1MOE	1069
						390030	39249	39260	TTTATAAAACTG	1-10-1MOE	1074
390030	39397	39408	TTTATAAAACTG	1-10-1MOE	1074
						401396	39488	39501	TGCAGGATGTTGAG	2-10-2MOE	945
147717	39545	39556	ATCTTCAGAGAT	1-10-1MOE	996
						147746	39641	39652	TAAAAACAACAA	1-10-1MOE	1073
147717	39693	39704	ATCTTCAGAGAT	1-10-1MOE	996
						147746	39729	39740	TAAAAACAACAA	1-10-1MOE	1073
147746	39877	39888	TAAAAACAACAA	1-10-1MOE	1073
						147746	40185	40196	TAAAAACAACAA	1-10-1MOE	1073
147746	40478	40489	TAAAAACAACAA	1-10-1MOE	1073
						398166	40589	40600	GGGCTTCTTCCA	1-10-1MOE	1070
147735	40662	40673	GGAGAAGCGCAG	1-10-1MOE	1016
						147746	40706	40717	TAAAAACAACAA	1-10-1MOE	1073

398166	40737	40748	GGGCTTCTTCCA	1-10-1MOE	1070
						147746	40854	40865	TAAAAACAACAA	1-10-1MOE	1073
401397	41012	41025	CTGGTCAGCATTGA	2-10-2MOE	946
						147718	41070	41081	TAATATGACTTG	1-10-1MOE	998
147718	41218	41229	TAATATGACTTG	1-10-1MOE	998
						147717	41221	41232	ATCTTCAGAGAT	1-10-1MOE	996
147717	41369	41380	ATCTTCAGAGAT	1-10-1MOE	996
						147717	41599	41610	ATCTTCAGAGAT	1-10-1MOE	996
147717	41747	41758	ATCTTCAGAGAT	1-10-1MOE	996
						401398	41768	41781	CAAAGTCCCTTAGC	2-10-2MOE	947
390030	42056	42067	TTTATAAAACTG	1-10-1MOE	1074
						398153	42157	42170	ATTTCTCTTACAGG	2-10-2MOE	948
398153	42305	42318	ATTTCTCTTACAGG	2-10-2MOE	948
						147710	42691	42702	TATAGCTCCTCT	1-10-1MOE	994
147079	43322	43333	TCCTTCCACTGA	1-10-1MOE	1001
						147080	43323	43334	CTCCTTCCACTG	1-10-1MOE	1021
147716	43477	43488	TTAACGAGCCTT	1-10-1MOE	949
						147746	43992	44003	TAAAAACAACAA	1-10-1MOE	1073
147736	44137	44148	AGGTAGGAGAAG	1-10-1MOE	963
						384545	44242	44253	CAAGTAGGATGT	1-10-1MOE	951
147687	44354	44365	CGACACGGGAAC	1-10-1MOE	950
						384545	44390	44401	CAAGTAGGATGT	1-10-1MOE	951
398110	44713	44726	GTTCCCTTTGCAGG	2-10-2MOE	952
						147705	45092	45103	CGGTTTTTGTTC	1-10-1MOE	1002
147705	45240	45251	CGGTTTTTGTTC	1-10-1MOE	1002
						147074	45977	45988	CCACTGATCCTG	1-10-1MOE	845
147075	45978	45989	TCCACTGATCCT	1-10-1MOE	1026
						147076	45979	45990	TTCCACTGATCC	1-10-1MOE	1029
147076	46127	46138	TTCCACTGATCC	1-10-1MOE	1029
						401399	46247	46260	ATTAGCCATATCTC	2-10-2MOE	953
147705	46555	46566	CGGTTTTTGTTC	1-10-1MOE	1002
						147714	46685	46696	TTCTGCTCCCAC	1-10-1MOE	986
147705	46703	46714	CGGTTTTTGTTC	1-10-1MOE	1002
						390030	46859	46870	TTTATAAAACTG	1-10-1MOE	1074
390030	46933	46944	TTTATAAAACTG	1-10-1MOE	1074
						147681	46984	46995	ATGTCATTAAAC	1-10-1MOE	965
390030	47007	47018	TTTATAAAACTG	1-10-1MOE	1074
						147746	47023	47034	TAAAAACAACAA	1-10-1MOE	1073
390030	47081	47092	TTTATAAAACTG	1-10-1MOE	1074
						147681	47132	47143	ATGTCATTAAAC	1-10-1MOE	965
147746	47171	47182	TAAAAACAACAA	1-10-1MOE	1073
						401400	47411	47424	AGCATTCAGCAGTG	2-10-2MOE	954
147746	47461	47472	TAAAAACAACAA	1-10-1MOE	1073
						147086	47608	47619	CTCTACACCAGG	1-10-1MOE	969
147087	47609	47620	CCTCTACACCAG	1-10-1MOE	982
						147088	47610	47621	CCCTCTACACCA	1-10-1MOE	1050
147090	47612	47623	TTCCCTCTACAC	1-10-1MOE	955
						147691	47729	47740	GAGGTGGGAAAA	1-10-1MOE	966
147086	47756	47767	CTCTACACCAGG	1-10-1MOE	969
						147088	47758	47769	CCCTCTACACCA	1-10-1MOE	1050
147089	47759	47770	TCCCTCTACACC	1-10-1MOE	956
						390030	47847	47858	TTTATAAAACTG	1-10-1MOE	1074
390030	47995	48006	TTTATAAAACTG	1-10-1MOE	1074
						147691	48393	48404	GAGGTGGGAAAA	1-10-1MOE	966

398147	48887	48900	CTACAGGACAATAC	2-10-2MOE	957
						147706	49133	49144	GCTGACATCTCG	1-10-1MOE	1071
147706	49281	49292	GCTGACATCTCG	1-10-1MOE	1071
						398168	49742	49753	TCGGACTTTGAA	1-10-1MOE	1008
401401	49791	49804	AACTGGGTTAAGTA	2-10-2MOE	958
						147689	49936	49947	CAGAGAAGGTCT	1-10-1MOE	987
401402	50192	50205	TGAACACGCTATCC	2-10-2MOE	959
						398117	50241	50254	TTTCCACTTGGGTG	2-10-2MOE	960
147736	50582	50593	AGGTAGGAGAAG	1-10-1MOE	963
						398168	50703	50714	TCGGACTTTGAA	1-10-1MOE	1008
398168	50849	50860	TCGGACTTTGAA	1-10-1MOE	1008
						147746	51019	51030	TAAAAACAACAA	1-10-1MOE	1073
147708	51101	51112	TTGATATAGTCA	1-10-1MOE	997
						147746	51178	51189	TAAAAACAACAA	1-10-1MOE	1073
147708	51247	51258	TTGATATAGTCA	1-10-1MOE	997
						147083	51281	51292	TACACCAGGTCA	1-10-1MOE	973
147081	51287	51298	GCTCCTTCCACT	1-10-1MOE	1006
						147082	51288	51299	AGCTCCTTCCAC	1-10-1MOE	1036
147746	51331	51342	TAAAAACAACAA	1-10-1MOE	1073
						147085	51416	51427	TCTACACCAGGT	1-10-1MOE	961
147083	51427	51438	TACACCAGGTCA	1-10-1MOE	973
						147081	51433	51444	GCTCCTTCCACT	1-10-1MOE	1006
147082	51434	51445	AGCTCCTTCCAC	1-10-1MOE	1036
						147728	51522	51533	GCCAGACAGAAG	1-10-1MOE	1013
147085	51562	51573	TCTACACCAGGT	1-10-1MOE	961
						147081	51633	51644	GCTCCTTCCACT	1-10-1MOE	1006
368360	51633	51646	AAGCTCCTTCCACT	2-10-2MOE	1035
						147082	51634	51645	AGCTCCTTCCAC	1-10-1MOE	1036
368361	51635	51648	GAAAGCTCCTTCCA	2-10-2MOE	962
						368360	51779	51792	AAGCTCCTTCCACT	2-10-2MOE	1035
147082	51780	51791	AGCTCCTTCCAC	1-10-1MOE	1036
						147736	51859	51870	AGGTAGGAGAAG	1-10-1MOE	963
147684	51867	51878	ACCCAGTCAGGG	1-10-1MOE	964
						147746	51918	51929	TAAAAACAACAA	1-10-1MOE	1073
147077	51988	51999	CTTCCACTGATC	1-10-1MOE	1047
						147746	52064	52075	TAAAAACAACAA	1-10-1MOE	1073
147084	52125	52136	CTACACCAGGTC	1-10-1MOE	993
						147079	52136	52147	TCCTTCCACTGA	1-10-1MOE	1001
147681	52231	52242	ATGTCATTAAAC	1-10-1MOE	965
						147084	52271	52282	CTACACCAGGTC	1-10-1MOE	993
147691	52312	52323	GAGGTGGGAAAA	1-10-1MOE	966
						401403	52318	52331	TTTCCTAGGAGGTG	2-10-2MOE	967
398167	52527	52538	CAGGCCATGTGG	1-10-1MOE	1059
						147703	52670	52681	TGGCTTCATGTC	1-10-1MOE	971
398167	52673	52684	CAGGCCATGTGG	1-10-1MOE	1059
						398165	52708	52719	GTTCTTAGGAAG	1-10-1MOE	968
398090	52708	52721	TTGTTCTTAGGAAG	2-10-2MOE	972
						147705	52716	52727	CGGTTTTTGTTC	1-10-1MOE	1002
147682	52717	52728	CGGGTACTATGG	1-10-1MOE	992
						398167	52762	52773	CAGGCCATGTGG	1-10-1MOE	1059
147703	52816	52827	TGGCTTCATGTC	1-10-1MOE	971
						398090	52854	52867	TTGTTCTTAGGAAG	2-10-2MOE	972
147704	52856	52867	TTGTTCTTAGGA	1-10-1MOE	1012
						147705	52862	52873	CGGTTTTTGTTC	1-10-1MOE	1002

398167	52908	52919	CAGGCCATGTGG	1-10-1MOE	1059
						147084	53704	53715	CTACACCAGGTC	1-10-1MOE	993
147088	53708	53719	CCCTCTACACCA	1-10-1MOE	1050
						147083	53849	53860	TACACCAGGTCA	1-10-1MOE	973
147084	53850	53861	CTACACCAGGTC	1-10-1MOE	993
						147086	53852	53863	CTCTACACCAGG	1-10-1MOE	969
147088	53854	53865	CCCTCTACACCA	1-10-1MOE	1050
						398167	53870	53881	CAGGCCATGTGG	1-10-1MOE	1059
147703	54137	54148	TGGCTTCATGTC	1-10-1MOE	971
						398155	54172	54185	TGTTTTTACACAGA	2-10-2MOE	970
390030	54263	54274	TTTATAAAACTG	1-10-1MOE	1074
						147705	54275	54286	CGGTTTTTGTTC	1-10-1MOE	1002
147703	54283	54294	TGGCTTCATGTC	1-10-1MOE	971
						390030	54409	54420	TTTATAAAACTG	1-10-1MOE	1074
147704	54965	54976	TTGTTCTTAGGA	1-10-1MOE	1012
						147705	54971	54982	CGGTTTTTGTTC	1-10-1MOE	1002
398090	55109	55122	TTGTTCTTAGGAAG	2-10-2MOE	972
						147705	55117	55128	CGGTTTTTGTTC	1-10-1MOE	1002
147083	55206	55217	TACACCAGGTCA	1-10-1MOE	973
						147084	55207	55218	CTACACCAGGTC	1-10-1MOE	993
147084	55353	55364	CTACACCAGGTC	1-10-1MOE	993
						147705	55524	55535	CGGTTTTTGTTC	1-10-1MOE	1002
147685	55602	55613	GGCTGACATTCA	1-10-1MOE	975
						401404	55638	55651	TGAGCTACAGTAGG	2-10-2MOE	974
147685	55748	55759	GGCTGACATTCA	1-10-1MOE	975
						147712	55819	55830	ACACCATCTCCC	1-10-1MOE	1005
147712	55965	55976	ACACCATCTCCC	1-10-1MOE	1005
						147707	56300	56311	TAGTCATTATCT	1-10-1MOE	977
147708	56306	56317	TTGATATAGTCA	1-10-1MOE	997
						390030	56321	56332	TTTATAAAACTG	1-10-1MOE	1074
147709	56326	56337	CCATTTTTATCA	1-10-1MOE	978
						398091	56333	56346	GGGCTTCTTCCATT	2-10-2MOE	979
401405	56408	56421	TGGTCAACTGAAAG	2-10-2MOE	976
						147707	56446	56457	TAGTCATTATCT	1-10-1MOE	977
147708	56452	56463	TTGATATAGTCA	1-10-1MOE	997
						147709	56472	56483	CCATTTTTATCA	1-10-1MOE	978
398091	56479	56492	GGGCTTCTTCCATT	2-10-2MOE	979
						401406	56570	56583	GGTGTGGATAACAG	2-10-2MOE	980
368366	56664	56677	CTGATCCTTAGAAG	2-10-2MOE	1019
						398148	57157	57170	TCATAACTATTAAG	2-10-2MOE	981
147082	57220	57231	AGCTCCTTCCAC	1-10-1MOE	1036
						398148	57303	57316	TCATAACTATTAAG	2-10-2MOE	981
147082	57366	57377	AGCTCCTTCCAC	1-10-1MOE	1036
						147743	57758	57769	AGGGCTTCCAGT	1-10-1MOE	1042
398093	57963	57976	TCGGACTTTGAAAA	2-10-2MOE	1009
						398093	58109	58122	TCGGACTTTGAAAA	2-10-2MOE	1009
147735	58279	58290	GGAGAAGCGCAG	1-10-1MOE	1016
						147087	58821	58832	CCTCTACACCAG	1-10-1MOE	982
147087	58967	58978	CCTCTACACCAG	1-10-1MOE	982
						390030	59180	59191	TTTATAAAACTG	1-10-1MOE	1074
390030	59326	59337	TTTATAAAACTG	1-10-1MOE	1074
						147711	59357	59368	AAGGGCCCTGGG	1-10-1MOE	1040
147743	59382	59393	AGGGCTTCCAGT	1-10-1MOE	1042
						147711	59503	59514	AAGGGCCCTGGG	1-10-1MOE	1040

147711	59675	59686	AAGGGCCCTGGG	1-10-1MOE	1040
						401407	59710	59723	CAGCTTAGGCAGAG	2-10-2MOE	983
147712	59711	59722	ACACCATCTCCC	1-10-1MOE	1005
						147713	59716	59727	CTCCCACACCAT	1-10-1MOE	985
147714	59721	59732	TTCTGCTCCCAC	1-10-1MOE	986
						147695	59722	59733	TCATTCCCCACT	1-10-1MOE	984
147715	59746	59757	GTTGAGCATGAC	1-10-1MOE	1077
						147711	59821	59832	AAGGGCCCTGGG	1-10-1MOE	1040
390030	59847	59858	TTTATAAAACTG	1-10-1MOE	1074
						147712	59857	59868	ACACCATCTCCC	1-10-1MOE	1005
147713	59862	59873	CTCCCACACCAT	1-10-1MOE	985
						147714	59867	59878	TTCTGCTCCCAC	1-10-1MOE	986
390030	59993	60004	TTTATAAAACTG	1-10-1MOE	1074
						389949	60471	60482	GCGCGAGCCCGA	1-10-1MOE	1061
147746	60619	60630	TAAAAACAACAA	1-10-1MOE	1073
						147689	61113	61124	CAGAGAAGGTCT	1-10-1MOE	987
398105	61267	61280	TGCACAGGCAGGTT	2-10-2MOE	1066
						147680	61473	61484	GTATGCACTGCT	1-10-1MOE	988
147080	61757	61768	CTCCTTCCACTG	1-10-1MOE	1021
						147078	61901	61912	CCTTCCACTGAT	1-10-1MOE	1044
147079	61902	61913	TCCTTCCACTGA	1-10-1MOE	1001
						147088	62215	62226	CCCTCTACACCA	1-10-1MOE	1050
401408	62600	62613	CAATGAAGCACAGG	2-10-2MOE	989
						147688	62843	62854	TCCCAAACAAAT	1-10-1MOE	990
147746	63102	63113	TAAAAACAACAA	1-10-1MOE	1073
						147746	63248	63259	TAAAAACAACAA	1-10-1MOE	1073
401409	63430	63443	ATTCTTAACACAGA	2-10-2MOE	991
						147682	63483	63494	CGGGTACTATGG	1-10-1MOE	992
147084	63677	63688	CTACACCAGGTC	1-10-1MOE	993
						147710	64847	64858	TATAGCTCCTCT	1-10-1MOE	994
147710	64993	65004	TATAGCTCCTCT	1-10-1MOE	994
						147746	65151	65162	TAAAAACAACAA	1-10-1MOE	1073
401410	65263	65276	CATTTAGGGTCTAA	2-10-2MOE	995
						147717	65862	65873	ATCTTCAGAGAT	1-10-1MOE	996
147717	65895	65906	ATCTTCAGAGAT	1-10-1MOE	996
						147708	65900	65911	TTGATATAGTCA	1-10-1MOE	997
147718	65909	65920	TAATATGACTTG	1-10-1MOE	998
						147717	66008	66019	ATCTTCAGAGAT	1-10-1MOE	996
147717	66041	66052	ATCTTCAGAGAT	1-10-1MOE	996
						147708	66046	66057	TTGATATAGTCA	1-10-1MOE	997
147718	66055	66066	TAATATGACTTG	1-10-1MOE	998
						401411	66123	66136	AGCCGCCTGAAGTG	2-10-2MOE	999
147697	66497	66508	CCCCAGCAGCGG	1-10-1MOE	1000
						368377	66562	66577	CTCCTTCCACTGATCC	3-10-3MOE	1030
147077	66563	66574	CTTCCACTGATC	1-10-1MOE	1047
						368358	66563	66576	TCCTTCCACTGATC	2-10-2MOE	1031
147078	66564	66575	CCTTCCACTGAT	1-10-1MOE	1044
						147079	66565	66576	TCCTTCCACTGA	1-10-1MOE	1001
147080	66566	66577	CTCCTTCCACTG	1-10-1MOE	1021
						147697	66643	66654	CCCCAGCAGCGG	1-10-1MOE	1000
368358	66709	66722	TCCTTCCACTGATC	2-10-2MOE	1031
						147078	66710	66721	CCTTCCACTGAT	1-10-1MOE	1044
147079	66711	66722	TCCTTCCACTGA	1-10-1MOE	1001
						147075	66999	67010	TCCACTGATCCT	1-10-1MOE	1026

147705	67067	67078	CGGTTTTTGTTC	1-10-1MOE	1002
						147088	67409	67420	CCCTCTACACCA	1-10-1MOE	1050
147080	67430	67441	CTCCTTCCACTG	1-10-1MOE	1021
						147082	67432	67443	AGCTCCTTCCAC	1-10-1MOE	1036
147737	67455	67466	ACAGCCAGGTAG	1-10-1MOE	1067
						147088	67555	67566	CCCTCTACACCA	1-10-1MOE	1050
147082	67578	67589	AGCTCCTTCCAC	1-10-1MOE	1036
						401412	67637	67650	TAAATCCTCTAGCA	2-10-2MOE	1003
147091	67729	67740	GTTCCCTCTACA	1-10-1MOE	1004
						147742	67737	67748	AACTTCAGTGTC	1-10-1MOE	1041
147712	68527	68538	ACACCATCTCCC	1-10-1MOE	1005
						147712	68673	68684	ACACCATCTCCC	1-10-1MOE	1005
147711	68760	68771	AAGGGCCCTGGG	1-10-1MOE	1040
						147711	68906	68917	AAGGGCCCTGGG	1-10-1MOE	1040
389965	69271	69282	CTGCAACATGAT	1-10-1MOE	1018
						389965	69417	69428	CTGCAACATGAT	1-10-1MOE	1018
368353	69519	69532	CACTGATCCTGCAC	2-10-2MOE	1007
						147080	69630	69641	CTCCTTCCACTG	1-10-1MOE	1021
147081	69631	69642	GCTCCTTCCACT	1-10-1MOE	1006
						368353	69665	69678	CACTGATCCTGCAC	2-10-2MOE	1007
398167	69757	69768	CAGGCCATGTGG	1-10-1MOE	1059
						398092	69758	69771	AGTCAGGCCATGTG	2-10-2MOE	1060
398093	69811	69824	TCGGACTTTGAAAA	2-10-2MOE	1009
						398168	69813	69824	TCGGACTTTGAA	1-10-1MOE	1008
398167	69903	69914	CAGGCCATGTGG	1-10-1MOE	1059
						398093	69957	69970	TCGGACTTTGAAAA	2-10-2MOE	1009
398094	70047	70060	ATCAGCCAGACAGA	2-10-2MOE	1010
						398095	70065	70078	CATCAGCAAGAGGC	2-10-2MOE	1011
147704	70137	70148	TTGTTCTTAGGA	1-10-1MOE	1012
						147728	70450	70461	GCCAGACAGAAG	1-10-1MOE	1013
398164	70464	70475	TTGTCGATCTGC	1-10-1MOE	1014
						398096	70562	70575	GGAGAAGCGCAGCT	2-10-2MOE	1015
147735	70564	70575	GGAGAAGCGCAG	1-10-1MOE	1016
						147737	70575	70586	ACAGCCAGGTAG	1-10-1MOE	1067
147735	70710	70721	GGAGAAGCGCAG	1-10-1MOE	1016
						147737	70721	70732	ACAGCCAGGTAG	1-10-1MOE	1067
404131	70729	70742	ACCTTCGATCACAG	2-10-2MOE	831
						368349	70762	70775	CTGCACTGACGAGT	2-10-2MOE	1017
389965	70930	70941	CTGCAACATGAT	1-10-1MOE	1018
						368366	70995	71008	CTGATCCTTAGAAG	2-10-2MOE	1019
368354	70999	71012	TCCACTGATCCTGC	2-10-2MOE	1024
						368375	71000	71015	CCTTCCACTGATCCTG	3-10-3MOE	1020
368356	71001	71014	CTTCCACTGATCCT	2-10-2MOE	1027
						368376	71001	71016	TCCTTCCACTGATCCT	3-10-3MOE	1028
368357	71002	71015	CCTTCCACTGATCC	2-10-2MOE	1046
						368377	71002	71017	CTCCTTCCACTGATCC	3-10-3MOE	1030
147077	71003	71014	CTTCCACTGATC	1-10-1MOE	1047
						368358	71003	71016	TCCTTCCACTGATC	2-10-2MOE	1031
368378	71003	71018	GCTCCTTCCACTGATC	3-10-3MOE	1032
						147078	71004	71015	CCTTCCACTGAT	1-10-1MOE	1044
368359	71005	71018	GCTCCTTCCACTGA	2-10-2MOE	1033
						368379	71005	71020	AAGCTCCTTCCACTGA	3-10-3MOE	1034
147080	71006	71017	CTCCTTCCACTG	1-10-1MOE	1021
						147082	71008	71019	AGCTCCTTCCAC	1-10-1MOE	1036

401413	71019	71032	TGCAGCCATGTACT	2-10-2MOE	1022
						147738	71067	71078	TGGGTGGCCGGG	1-10-1MOE	1069
147739	71071	71082	CGTTTGGGTGGC	1-10-1MOE	1023
						147741	71129	71140	CACCCACTGGTG	1-10-1MOE	1055
368354	71145	71158	TCCACTGATCCTGC	2-10-2MOE	1024
						368355	71146	71159	TTCCACTGATCCTG	2-10-2MOE	1025
147075	71147	71158	TCCACTGATCCT	1-10-1MOE	1026
						368356	71147	71160	CTTCCACTGATCCT	2-10-2MOE	1027
368376	71147	71162	TCCTTCCACTGATCCT	3-10-3MOE	1028
						147076	71148	71159	TTCCACTGATCC	1-10-1MOE	1029
368357	71148	71161	CCTTCCACTGATCC	2-10-2MOE	1046
						368377	71148	71163	CTCCTTCCACTGATCC	3-10-3MOE	1030
147077	71149	71160	CTTCCACTGATC	1-10-1MOE	1047
						368358	71149	71162	TCCTTCCACTGATC	2-10-2MOE	1031
368378	71149	71164	GCTCCTTCCACTGATC	3-10-3MOE	1032
						147078	71150	71161	CCTTCCACTGAT	1-10-1MOE	1044
368359	71151	71164	GCTCCTTCCACTGA	2-10-2MOE	1033
						368379	71151	71166	AAGCTCCTTCCACTGA	3-10-3MOE	1034
368360	71153	71166	AAGCTCCTTCCACT	2-10-2MOE	1035
						147082	71154	71165	AGCTCCTTCCAC	1-10-1MOE	1036
368381	71155	71170	GGGAAAGCTCCTTCCA	3-10-3MOE	1037
						390030	71986	71997	TTTATAAAACTG	1-10-1MOE	1074
390030	72132	72143	TTTATAAAACTG	1-10-1MOE	1074
						147711	72300	72311	AAGGGCCCTGGG	1-10-1MOE	1040
401414	72347	72360	TTGCAATGTCTGGC	2-10-2MOE	1038
						147741	72400	72411	CACCCACTGGTG	1-10-1MOE	1055
401415	72415	72428	GATTTATCTGGCTG	2-10-2MOE	1039
						147711	72446	72457	AAGGGCCCTGGG	1-10-1MOE	1040
147742	72575	72586	AACTTCAGTGTC	1-10-1MOE	1041
						147743	72690	72701	AGGGCTTCCAGT	1-10-1MOE	1042
147744	72694	72705	AGGAAGGGCTTC	1-10-1MOE	1043
						147745	72700	72711	TTGACCAGGAAG	1-10-1MOE	1058
147742	72721	72732	AACTTCAGTGTC	1-10-1MOE	1041
						147743	72836	72847	AGGGCTTCCAGT	1-10-1MOE	1042
147744	72840	72851	AGGAAGGGCTTC	1-10-1MOE	1043
						368357	72898	72911	CCTTCCACTGATCC	2-10-2MOE	1046
147078	72900	72911	CCTTCCACTGAT	1-10-1MOE	1044
						398157	72903	72916	GGAAACATACCCTG	2-10-2MOE	1045
368357	73044	73057	CCTTCCACTGATCC	2-10-2MOE	1046
						147077	73045	73056	CTTCCACTGATC	1-10-1MOE	1047
147746	73052	73063	TAAAAACAACAA	1-10-1MOE	1073
						147746	73101	73112	TAAAAACAACAA	1-10-1MOE	1073
398160	73139	73152	GAATAGGTTAAGGC	2-10-2MOE	1048
						147746	73198	73209	TAAAAACAACAA	1-10-1MOE	1073
398161	73238	73251	AACAATGTGTTGTA	2-10-2MOE	1049
						147088	73419	73430	CCCTCTACACCA	1-10-1MOE	1050
404140	73457	73470	GCACACAGCTGAGG	2-10-2MOE	1051
						404139	73459	73472	GTGCACACAGCTGA	2-10-2MOE	1052
399301	73461	73474	GTGTGCACACAGCT	2-10-2MOE	1542
						404137	73463	73476	CAGTGTGCACACAG	2-10-2MOE	1053
404138	73465	73478	CTCAGTGTGCACAC	2-10-2MOE	1054
						147741	73705	73716	CACCCACTGGTG	1-10-1MOE	1055
404135	73858	73871	CATTTCCATGGCCA	2-10-2MOE	1056
						398167	74008	74019	CAGGCCATGTGG	1-10-1MOE	1059

398092	74009	74022	AGTCAGGCCATGTG	2-10-2MOE	1060
						398162	74114	74127	ACCAAACAGTTCAG	2-10-2MOE	1057
147745	74137	74148	TTGACCAGGAAG	1-10-1MOE	1058
						398167	74154	74165	CAGGCCATGTGG	1-10-1MOE	1059
398092	74155	74168	AGTCAGGCCATGTG	2-10-2MOE	1060
						389949	74310	74321	GCGCGAGCCCGA	1-10-1MOE	1061
147740	74485	74496	TGTGAGGCTCCA	1-10-1MOE	1062
						389950	74527	74538	CCCTGAAGGTTC	1-10-1MOE	1063
398101	74656	74669	TTTGATAAAGCCCT	2-10-2MOE	1064
						398104	74805	74818	CAAGAAGACCTTAC	2-10-2MOE	1065
147737	74893	74904	ACAGCCAGGTAG	1-10-1MOE	1067
						398105	74894	74907	TGCACAGGCAGGTT	2-10-2MOE	1066
147737	74919	74930	ACAGCCAGGTAG	1-10-1MOE	1067
						398106	74974	74987	TGGAAAACTGCACC	2-10-2MOE	1068
404199	75045	75058	GGTCATGCACAGGC	2-10-2MOE	867
						404134	75048	75061	TCAGGTCATGCACA	2-10-2MOE	873
398106	75120	75133	TGGAAAACTGCACC	2-10-2MOE	1068
						147738	75155	75166	TGGGTGGCCGGG	1-10-1MOE	1069
404132	75227	75240	CCTTGGAATGTCTG	2-10-2MOE	852
						147738	75301	75312	TGGGTGGCCGGG	1-10-1MOE	1069
398166	75499	75510	GGGCTTCTTCCA	1-10-1MOE	1070
						147746	75617	75628	TAAAAACAACAA	1-10-1MOE	1073
147706	75686	75697	GCTGACATCTCG	1-10-1MOE	1071
						398112	75730	75743	CAGCCTGGCACCTA	2-10-2MOE	1072
147746	75763	75774	TAAAAACAACAA	1-10-1MOE	1073
						398115	75786	75799	AGTAAATATTGGCT	2-10-2MOE	1076
390030	75839	75850	TTTATAAAACTG	1-10-1MOE	1074
						398114	75916	75929	AGGCATATAGCAGA	2-10-2MOE	1075
398115	75932	75945	AGTAAATATTGGCT	2-10-2MOE	1076
						404133	75968	75981	TATTCCATGGCCAT	2-10-2MOE	872
147715	77045	77056	GTTGAGCATGAC	1-10-1MOE	1077
						147715	77190	77201	GTTGAGCATGAC	1-10-1MOE	1077
147693	77385	77396	GTGCGCTCCCAT	1-10-1MOE	1078
						398173	40201	40212	CAGCCTGGGCAC	1-10-1MOE	1543
398173	72764	72775	CA GCCTGGGCA C	1-10-1MOE	1543
						399096	1986	1999	TGCTCGAACTCCTT	2-10-2MOE	1544
399102	52822	52835	GAAGTCACTGGCTT	2-10-2MOE	1545
						399103	52824	52837	GGGAAGTCACTGGC	2-10-2MOE	1546
399113	59827	59840	GTTAGGCAAAGGGC	2-10-2MOE	1547
						399132	69977	69990	GGGCTGAGTGACCC	2-10-2MOE	1548
399173	74592	74605	ATGCTAGTGCACTA	2-10-2MOE	1549
						399208	75900	75913	AGCTCGCTACCTCT	2-10-2MOE	1550
399276	27559	27572	GAGGTATCCCATCT	2-10-2MOE	1551
						399315	74039	74052	GGCAACTTCAACCT	2-10-2MOE	1552

Table 19: targeting SEQ ID NO:12 and have the short antisense compounds of 1 or 2 mispairing

ISISNO.	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SeqID NO
						398163	20	31	ATGTCAACCGGC	1-10-1MOE	908
384545	23	34	CAAGTAGGATGT	1-10-1MOE	951

147733	26	37	TTCTTGATGTCC	1-10-1MOE	891
						147721	59	70	AATGCAGGATCT	1-10-1MOE	1118
147700	110	121	GCGCTAGGCCGC	1-10-1MOE	1110
						384545	130	141	CAAGTAGGATGT	1-10-1MOE	951
147705	159	170	CGGTTTTTGTTC	1-10-1MOE	1002
						147701	167	178	CCATGGCGGGAC	1-10-1MOE	921
398164	198	209	TTGTCGATCTGC	1-10-1MOE	1014
						147730	199	210	CTTGTCCATCAG	1-10-1MOE	1121
147702	226	237	CTGGTAAATAGC	1-10-1MOE	898
						147703	245	256	TGGCTTCATGTC	1-10-1MOE	971
147705	266	277	CGGTTTTTGTTC	1-10-1MOE	1002
						398165	283	294	GTTCTTAGGAAG	1-10-1MOE	968
147704	285	296	TTGTTCTTAGGA	1-10-1MOE	1012
						147705	291	302	CGGTTTTTGTTC	1-10-1MOE	1002
147709	311	322	CCATTTTTATCA	1-10-1MOE	978
						147733	349	360	TTCTTGATGTCC	1-10-1MOE	891
147707	360	371	TAGTCATTATCT	1-10-1MOE	977
						147708	366	377	TTGATATAGTCA	1-10-1MOE	997
390030	381	392	TTTATAAAACTG	1-10-1MOE	1074
						147709	386	397	CCATTTTTATCA	1-10-1MOE	978
147081	393	404	GCTCCTTCCACT	1-10-1MOE	1006
						398091	393	406	GGGCTTCTTCCATT	2-10-2MOE	979
398166	395	406	GGGCTTCTTCCA	1-10-1MOE	1070
						147712	461	472	ACACCATCTCCC	1-10-1MOE	1005
147713	466	477	CTCCCACACCAT	1-10-1MOE	985
						147714	471	482	TTCTGCTCCCAC	1-10-1MOE	986
147710	502	513	TATAGCTCCTCT	1-10-1MOE	994
						147736	551	562	AGGTAGGAGAAG	1-10-1MOE	963
147717	574	585	ATCTTCAGAGAT	1-10-1MOE	996
						147717	607	618	ATCTTCAGAGAT	1-10-1MOE	996
147710	609	620	TATAGCTCCTCT	1-10-1MOE	994
						147708	612	623	TTGATATAGTCA	1-10-1MOE	997
147718	621	632	TAATATGACTTG	1-10-1MOE	998
						147746	625	636	TAAAAACAACAA	1-10-1MOE	1073
147736	658	669	AGGTAGGAGAAG	1-10-1MOE	963
						147720	676	687	GATCTCTCGAGT	1-10-1MOE	1117
147721	683	694	AATGCAGGATCT	1-10-1MOE	1118
						398167	704	715	CAGGCCATGTGG	1-10-1MOE	1059
398092	705	718	AGTCAGGCCATGTG	2-10-2MOE	1060
						147722	709	720	AAAGTCAGGCCA	1-10-1MOE	1130
147723	715	726	GACTCCAAAGTC	1-10-1MOE	892
						147746	733	744	TAAAAACAACAA	1-10-1MOE	1073
398093	758	771	TCGGACTTTGAAAA	2-10-2MOE	1009
						398168	760	771	TCGGACTTTGAA	1-10-1MOE	1008
147725	761	772	CTCGGACTTTGA	1-10-1MOE	1119
						147726	766	777	TGACTCTCGGAC	1-10-1MOE	1120
147738	780	791	TGGGTGGCCGGG	1-10-1MOE	1069
						147727	807	818	CAGTGGACCACA	1-10-1MOE	1128
147728	846	857	GCCAGACAGAAG	1-10-1MOE	1013
						398094	848	861	ATCAGCCAGACAGA	2-10-2MOE	1010
398169	849	860	TCAGCCAGACAG	1-10-1MOE	909
						147729	863	874	GTAAGAGGCAGG	1-10-1MOE	920
398095	866	879	CATCAGCAAGAGGC	2-10-2MOE	1011
						398164	873	884	TTGTCGATCTGC	1-10-1MOE	1014

147730	874	885	CTTGTCCATCAG	1-10-1MOE	1121
						147731	880	891	TTTCCTCTTGTC	1-10-1MOE	934
147732	885	896	GGGTCTTTCCTC	1-10-1MOE	1122
						147738	888	899	TGGGTGGCCGGG	1-10-1MOE	1069
147733	906	917	TTCTTGATGTCC	1-10-1MOE	891
						398096	971	984	GGAGAAGCGCAGCT	2-10-2MOE	1015
147735	973	984	GGAGAAGCGCAG	1-10-1MOE	1016
						147736	978	989	AGGTAGGAGAAG	1-10-1MOE	963
147729	979	990	GTAAGAGGCAGG	1-10-1MOE	920
						147737	984	995	ACAGCCAGGTAG	1-10-1MOE	1067
368349	1025	1038	CTGCACTGACGAGT	2-10-2MOE	1017
						368369	1025	1040	TCCTGCACTGACGAGT	3-10-3MOE	893
368350	1027	1040	TCCTGCACTGACGA	2-10-2MOE	1079
						368370	1027	1042	GATCCTGCACTGACGA	3-10-3MOE	1080
368351	1029	1042	GATCCTGCACTGAC	2-10-2MOE	1081
						368371	1029	1044	CTGATCCTGCACTGAC	3-10-3MOE	1082
368352	1031	1044	CTGATCCTGCACTG	2-10-2MOE	1105
						368372	1031	1046	CACTGATCCTGCACTG	3-10-3MOE	894
368353	1033	1046	CACTGATCCTGCAC	2-10-2MOE	1007
						368373	1033	1048	TCCACTGATCCTGCAC	3-10-3MOE	1083
368354	1035	1048	TCCACTGATCCTGC	2-10-2MOE	1024
						368368	1035	1048	TCCACTGATCCTTA	2-10-2MOE	1127
368374	1035	1050	CTTCCACTGATCCTGC	3-10-3MOE	1126
						368388	1035	1050	CTTCCACTGATCCTTA	3-10-3MOE	895
147074	1036	1047	CCACTGATCCTG	1-10-1MOE	845
						368355	1036	1049	TTCCACTGATCCTG	2-10-2MOE	1025
368375	1036	1051	CCTTCCACTGATCCTG	3-10-3MOE	1020
						147075	1037	1048	TCCACTGATCCT	1-10-1MOE	1026
368356	1037	1050	CTTCCACTGATCCT	2-10-2MOE	1027
						368376	1037	1052	TCCTTCCACTGATCCT	3-10-3MOE	1028
147076	1038	1049	TTCCACTGATCC	1-10-1MOE	1029
						368357	1038	1051	CCTTCCACTGATCC	2-10-2MOE	1046
368377	1038	1053	CTCCTTCCA CTGATCC	3-10-3MOE	1030
						147077	1039	1050	CTTCCACTGATC	1-10-1MOE	1047
368358	1039	1052	TCCTTCCACTGATC	2-10-2MOE	1031
						368378	1039	1054	GCTCCTTCCACTGATC	3-10-3MOE	1032
147078	1040	1051	CCTTCCACTGAT	1-10-1MOE	1044
						147079	1041	1052	TCCTTCCACTGA	1-10-1MOE	1001
368359	1041	1054	GCTCCTTCCACTGA	2-10-2MOE	1033
						368379	1041	1056	AAGCTCCTTCCACTGA	3-10-3MOE	1034
147080	1042	1053	CTCCTTCCACTG	1-10-1MOE	1021
						147081	1043	1054	GCTCCTTCCACT	1-10-1MOE	1006
368360	1043	1056	AAGCTCCTTCCACT	2-10-2MOE	1035
						368380	1043	1058	GAAAGCTCCTTCCACT	3-10-3MOE	896
147082	1044	1055	AGCTCCTTCCAC	1-10-1MOE	1036
						368361	1045	1058	GAAAGCTCCTTCCA	2-10-2MOE	962
368381	1045	1060	GGGAAAGCTCCTTCCA	3-10-3MOE	1037
						147729	1087	1098	GTAAGAGGCAGG	1-10-1MOE	920
147738	1103	1114	TGGGTGGCCGGG	1-10-1MOE	1069
						147739	1107	1118	CGTTTGGGTGGC	1-10-1MOE	1023
147740	1124	1135	TGTGAGGCTCCA	1-10-1MOE	1062
						398117	1164	1177	TTTCCACTTGGGTG	2-10-2MOE	960
147741	1165	1176	CACCCACTGGTG	1-10-1MOE	1055
						398097	1194	1207	GGCAGTCTTTATCC	2-10-2MOE	897

398098	1272	1285	TAACTTCAGTGTCT	2-10-2MOE	1131
						398117	1272	1285	TTTCCACTTGGGTG	2-10-2MOE	960
147742	1273	1284	AACTTCAGTGTC	1-10-1MOE	1041
						147698	1293	1304	CCCGCCACCACC	1-10-1MOE	928
147743	1388	1399	AGGGCTTCCAGT	1-10-1MOE	1042
						398099	1388	1401	GAAGGGCTTCCAGT	2-10-2MOE	1132
147744	1392	1403	AGGAAGGGCTTC	1-10-1MOE	1043
						398100	1395	1408	TGACCAGGAAGGGC	2-10-2MOE	1133
147745	1398	1409	TTGACCAGGAAG	1-10-1MOE	1058
						398157	1455	1468	GGAAACATACCCTG	2-10-2MOE	1045
147745	1458	1469	TTGACCAGGAAG	1-10-1MOE	1058
						398167	1475	1486	CAGGCCATGTGG	1-10-1MOE	1059
398118	1564	1577	CGCGAGATATCTAA	2-10-2MOE	1084
						147697	1575	1586	CCCCAGCAGCGG	1-10-1MOE	1000
147076	1596	1607	TTCCACTGATCC	1-10-1MOE	1029
						368357	1596	1609	CCTTCCACTGATCC	2-10-2MOE	1046
147077	1597	1608	CTTCCACTGATC	1-10-1MOE	1047
						147078	1598	1609	CCTTCCACTGAT	1-10-1MOE	1044
398118	1672	1685	CGCGAGATATCTAA	2-10-2MOE	1084
						398158	1681	1694	AGGCCCTGAGATTA	2-10-2MOE	1134
147697	1683	1694	CCCCAGCAGCGG	1-10-1MOE	1000
						398159	1686	1699	GGTTAAGGCCCTGA	2-10-2MOE	1135
398160	1691	1704	GAATAGGTTAAGGC	2-10-2MOE	1048
						398163	1711	1722	ATGTCAACCGGC	1-10-1MOE	908
147733	1717	1728	TTCTTGATGTCC	1-10-1MOE	891
						147089	1747	1758	TCCCTCTACACC	1-10-1MOE	956
147090	1748	1759	TTCCCTCTACAC	1-10-1MOE	955
						147746	1750	1761	TAAAAACAACAA	1-10-1MOE	1073
389949	1777	1788	GCGCGAGCCCGA	1-10-1MOE	1061
						398161	1790	1803	AACAATGTGTTGTA	2-10-2MOE	1049
147746	1799	1810	TAAAAACAACAA	1-10-1MOE	1073
						147700	1801	1812	GCGCTAGGCCGC	1-10-1MOE	1110
147740	1806	1817	TGTGAGGCTCCA	1-10-1MOE	1062
						398163	1819	1830	ATGTCAACCGGC	1-10-1MOE	908
147733	1825	1836	TTCTTGATGTCC	1-10-1MOE	891
						389950	1848	1859	CCCTGAAGGTTC	1-10-1MOE	1063
147701	1858	1869	CCATGGCGGGAC	1-10-1MOE	921
						398164	1889	1900	TTGTCGATCTGC	1-10-1MOE	1014
147730	1890	1901	CTTGTCCATCAG	1-10-1MOE	1121
						147700	1909	1920	GCGCTAGGCCGC	1-10-1MOE	1110
398119	1920	1933	CGCACCTGGTAAAT	2-10-2MOE	1085
						147685	1957	1968	GGCTGACATTCA	1-10-1MOE	975
147701	1966	1977	CCATGGCGGGAC	1-10-1MOE	921
						398120	1966	1979	GTTCAAGCGGCCTA	2-10-2MOE	1086
398101	1977	1990	TTTGATAAAGCCCT	2-10-2MOE	1064
						398164	1997	2008	TTGTCGATCTGC	1-10-1MOE	1014
147730	1998	2009	CTTGTCCATCAG	1-10-1MOE	1121
						147702	2025	2036	CTGGTAAATAGC	1-10-1MOE	898
398119	2028	2041	CGCACCTGGTAAAT	2-10-2MOE	1085
						398120	2074	2087	GTTCAAGCGGCCTA	2-10-2MOE	1086
398105	2099	2112	TGCACAGGCAGGTT	2-10-2MOE	1066
						147736	2204	2215	AGGTAGGAGAAG	1-10-1MOE	963
147741	2257	2268	CACCCACTGGTG	1-10-1MOE	1055
						398104	2272	2285	CAAGAAGACCTTAC	2-10-2MOE	1065

147737	2360	2371	ACAGCCAGGTAG	1-10-1MOE	1067
						398105	2361	2374	TGCACAGGCAGGTT	2-10-2MOE	1066
147737	2386	2397	ACAGCCAGGTAG	1-10-1MOE	1067
						398095	2407	2420	CATCAGCAAGAGGC	2-10-2MOE	1011
398106	2441	2454	TGGAAAACTGCACC	2-10-2MOE	1068
						398107	2447	2460	TATTCCTGGAAAAC	2-10-2MOE	902
398121	2474	2487	GTGCCTAGCACAGA	2-10-2MOE	1097
						147745	2497	2508	TTGACCAGGAAG	1-10-1MOE	1058
147712	2499	2510	ACACCATCTCCC	1-10-1MOE	1005
						398108	2544	2557	GGAATGTCTGAGTT	2-10-2MOE	1136
147691	2575	2586	GAGGTGGGAAAA	1-10-1MOE	966
						398121	2582	2595	GTGCCTAGCACAGA	2-10-2MOE	1097
147738	2622	2633	TGGGTGGCCGGG	1-10-1MOE	1069
						398162	2666	2679	ACCAAACAGTTCAG	2-10-2MOE	1057
147745	2689	2700	TTGACCAGGAAG	1-10-1MOE	1058
						398167	2706	2717	CAGGCCATGTGG	1-10-1MOE	1059
398092	2707	2720	AGTCAGGCCATGTG	2-10-2MOE	1060
						398109	2714	2727	CAAGAAGTGTGGTT	2-10-2MOE	903
398110	2852	2865	GTTCCCTTTGCA GG	2-10-2MOE	952
						147091	2854	2865	GTTCCCTCTACA	1-10-1MOE	1004
147723	2924	2935	GACTCCAAAGTC	1-10-1MOE	892
						398111	2937	2950	GTGAAAATGCTGGC	2-10-2MOE	904
398166	2966	2977	GGGCTTCTTCCA	1-10-1MOE	1070
						147089	2978	2989	TCCCTCTACACC	1-10-1MOE	956
147090	2979	2990	TTCCCTCTACAC	1-10-1MOE	955
						147706	3007	3018	GCTGACATCTCG	1-10-1MOE	1071
389949	3008	3019	GCGCGAGCCCGA	1-10-1MOE	1061
						147723	3032	3043	GACTCCAAAGTC	1-10-1MOE	892
147740	3037	3048	TGTGAGGCTCCA	1-10-1MOE	1062
						398112	3051	3064	CAGCCTGGCACCTA	2-10-2MOE	1072
389950	3079	3090	CCCTGAAGGTTC	1-10-1MOE	1063
						147746	3084	3095	TAAAAACAACAA	1-10-1MOE	1073
398122	3148	3161	CCCTTTACACAAGT	2-10-2MOE	1087
						147089	3151	3162	TCCCTCTACACC	1-10-1MOE	956
147090	3152	3163	TTCCCTCTACAC	1-10-1MOE	955
						398113	3160	3173	AGGAGGTTAAACCA	2-10-2MOE	905
147685	3188	3199	GGCTGACATTCA	1-10-1MOE	975
						398101	3208	3221	TTTGATAAAGCCCT	2-10-2MOE	1064
398102	3234	3247	CTACCTGAGGATTT	2-10-2MOE	899
						398123	3235	3248	CTCAAAATAGATTT	2-10-2MOE	1088
398114	3237	3250	AGGCATATAGCAGA	2-10-2MOE	1075
						398103	3241	3254	CCCAGTACTACCTG	2-10-2MOE	900
398115	3253	3266	AGTAAATATTGGCT	2-10-2MOE	1076
						398122	3256	3269	CCCTTTACACAAGT	2-10-2MOE	1087
147089	3259	3270	TCCCTCTACACC	1-10-1MOE	956
						147090	3260	3271	TTCCCTCTACAC	1-10-1MOE	955
398116	3266	3279	TAATGACCTGATGA	2-10-2MOE	1137
						390030	3306	3317	TTTATAAAACTG	1-10-1MOE	1074
398123	3343	3356	CTCAAAATAGATTT	2-10-2MOE	1088
						147736	3435	3446	AGGTAGGAGAAG	1-10-1MOE	963
398104	3503	3516	CAAGAAGACCTTAC	2-10-2MOE	1065
						147737	3591	3602	ACAGCCAGGTAG	1-10-1MOE	1067
398105	3592	3605	TGCACAGGCAGGTT	2-10-2MOE	1066
						147719	3608	3619	CCAACTCCAACT	1-10-1MOE	1116

147737	3617	3628	ACAGCCAGGTAG	1-10-1MOE	1067
						401398	3621	3634	CAAAGTCCCTTAGC	2-10-2MOE	947
147079	3637	3648	TCCTTCCACTGA	1-10-1MOE	1001
						147080	3638	3649	CTCCTTCCACTG	1-10-1MOE	1021
398095	3638	3651	CATCAGCAAGAGGC	2-10-2MOE	1011
						398106	3672	3685	TGGAAAACTGCACC	2-10-2MOE	1068
147733	3687	3698	TTCTTGATGTCC	1-10-1MOE	891
						147731	3688	3699	TTTCCTCTTGTC	1-10-1MOE	934
147719	3716	3727	CCAACTCCAACT	1-10-1MOE	1116
						147745	3728	3739	TTGACCAGGAAG	1-10-1MOE	1058
147683	3740	3751	GCTTACGATTGT	1-10-1MOE	922
						147079	3745	3756	TCCTTCCACTGA	1-10-1MOE	1001
147080	3746	3757	CTCCTTCCACTG	1-10-1MOE	1021
						398108	3775	3788	GGAATGTCTGAGTT	2-10-2MOE	1136
147733	3795	3806	TTCTTGATGTCC	1-10-1MOE	891
						147731	3796	3807	TTTCCTCTTGTC	1-10-1MOE	934
147691	3806	3817	GAGGTGGGAAAA	1-10-1MOE	966
						147738	3853	3864	TGGGTGGCCGGG	1-10-1MOE	1069
398167	3926	3937	CAGGCCATGTGG	1-10-1MOE	1059
						147691	3978	3989	GAGGTGGGAAAA	1-10-1MOE	966
398167	4034	4045	CAGGCCATGTGG	1-10-1MOE	1059
						147091	4085	4096	GTTCCCTCTACA	1-10-1MOE	1004
147691	4086	4097	GAGGTGGGAAAA	1-10-1MOE	966
						398111	4168	4181	GTGAAAATGCTGGC	2-10-2MOE	904
398166	4197	4208	GGGCTTCTTCCA	1-10-1MOE	1070
						147091	4223	4234	GTTCCCTCTACA	1-10-1MOE	1004
147092	4224	4235	TGTTCCCTCTAC	1-10-1MOE	901
						398112	4282	4295	CAGCCTGGCACCTA	2-10-2MOE	1072
147746	4315	4326	TAAAAACAACAA	1-10-1MOE	1073
						398113	4391	4404	AGGAGGTTAAACCA	2-10-2MOE	905
147723	4422	4433	GACTCCAAAGTC	1-10-1MOE	892
						398114	4468	4481	AGGCATATAGCAGA	2-10-2MOE	1075
398115	4484	4497	AGTAAATATTGGCT	2-10-2MOE	1076
						390030	4491	4502	TTTATAAAACTG	1-10-1MOE	1074
398116	4497	4510	TAATGACCTGATGA	2-10-2MOE	1137
						147723	4530	4541	GACTCCAAAGTC	1-10-1MOE	892
390030	4599	4610	TTTATAAAACTG	1-10-1MOE	1074
						398124	4761	4774	CACATGAGCTATTC	2-10-2MOE	1089
398124	4869	4882	CACATGAGCTATTC	2-10-2MOE	1089
						147703	4926	4937	TGGCTTCATGTC	1-10-1MOE	971
147692	4928	4939	CTCACCTTCATG	1-10-1MOE	1113
						147696	4975	4986	TGGATGATTGGC	1-10-1MOE	906
147703	5034	5045	TGGCTTCATGTC	1-10-1MOE	971
						147692	5036	5047	CTCACCTTCATG	1-10-1MOE	1113
147098	5173	5184	AGTTGTTGTTCC	1-10-1MOE	1112
						398125	5183	5196	CAGTAAGGAATTTT	2-10-2MOE	913
398126	5216	5229	GTGAAGTGAGTCAT	2-10-2MOE	1090
						147098	5281	5292	AGTTGTTGTTCC	1-10-1MOE	1112
398127	5283	5296	GGTCACTCAAGATG	2-10-2MOE	1091
						398126	5324	5337	GTGAAGTGAGTCAT	2-10-2MOE	1090
398128	5335	5348	CTAAATTTAGTTCA	2-10-2MOE	911
						398127	5391	5404	GGTCACTCAAGATG	2-10-2MOE	1091
398128	5443	5456	CTAAATTTAGTTCA	2-10-2MOE	911
						147712	5474	5485	ACACCATCTCCC	1-10-1MOE	1005

147736	5600	5611	AGGTAGGAGAAG	1-10-1MOE	963
						147746	5606	5617	TAAAAACAACAA	1-10-1MOE	1073
398129	5628	5641	TTTGAGGAGCTATT	2-10-2MOE	1106
						147085	5654	5665	TCTACACCAGGT	1-10-1MOE	961
147736	5708	5719	AGGTAGGAGAAG	1-10-1MOE	963
						398129	5736	5749	TTTGAGGAGCTATT	2-10-2MOE	1106
147679	5934	5945	CAAAAGGATCCC	1-10-1MOE	907
						147723	6229	6240	GACTCCAAAGTC	1-10-1MOE	892
147723	6338	6349	GACTCCAAAGTC	1-10-1MOE	892
						390030	6803	6814	TTTATAAAACTG	1-10-1MOE	1074
398142	6885	6898	CCAGCACACTGGAA	2-10-2MOE	923
						390030	6912	6923	TTTATAAAACTG	1-10-1MOE	1074
398142	6994	7007	CCAGCACACTGGAA	2-10-2MOE	923
						147695	7054	7065	TCATTCCCCACT	1-10-1MOE	984
147695	7163	7174	TCATTCCCCACT	1-10-1MOE	984
						398166	7197	7208	GGGCTTCTTCCA	1-10-1MOE	1070
398166	7306	7317	GGGCTTCTTCCA	1-10-1MOE	1070
						147684	7442	7453	ACCCAGTCAGGG	1-10-1MOE	964
398130	7694	7707	TTAGTATGACAGCT	2-10-2MOE	925
						398131	7711	7724	GGACTCACTCAGCA	2-10-2MOE	1092
398130	7802	7815	TTAGTATGACAGCT	2-10-2MOE	925
						398125	7804	7817	CAGTAAGGAATTTT	2-10-2MOE	913
398131	7819	7832	GGACTCACTCAGCA	2-10-2MOE	1092
						390030	7877	7888	TTTATAAAACTG	1-10-1MOE	1074
398125	7912	7925	CAGTAAGGAATTTT	2-10-2MOE	913
						390030	7985	7996	TTTATAAAACTG	1-10-1MOE	1074
398132	8031	8044	TCAGGGCTACTCAT	2-10-2MOE	1093
						398132	8139	8152	TCAGGGCTACTCAT	2-10-2MOE	1093
147684	8148	8159	ACCCAGTCAGGG	1-10-1MOE	964
						147684	8256	8267	ACCCAGTCAGGG	1-10-1MOE	964
398163	8365	8376	ATGTCAACCGGC	1-10-1MOE	908
						398166	8447	8458	GGGCTTCTTCCA	1-10-1MOE	1070
398163	8473	8484	ATGTCAACCGGC	1-10-1MOE	908
						398166	8555	8566	GGGCTTCTTCCA	1-10-1MOE	1070
147718	8631	8642	TAATATGACTTG	1-10-1MOE	998
						147691	8698	8709	GAGGTGGGAAAA	1-10-1MOE	966
147691	8806	8817	GAGGTGGGAAAA	1-10-1MOE	966
						147728	8835	8846	GCCAGACAGAAG	1-10-1MOE	1013
147727	8876	8887	CAGTGGACCACA	1-10-1MOE	1128
						147728	8943	8954	GCCAGACAGAAG	1-10-1MOE	1013
398169	8946	8957	TCAGCCAGACAG	1-10-1MOE	909
						147727	8984	8995	CAGTGGACCACA	1-10-1MOE	1128
147742	9060	9071	AACTTCAGTGTC	1-10-1MOE	1041
						398133	9112	9125	CAGCACTAGATTCA	2-10-2MOE	1094
384545	9135	9146	CAAGTAGGATGT	1-10-1MOE	951
						147742	9168	9179	AACTTCAGTGTC	1-10-1MOE	1041
398133	9220	9233	CAGCACTAGATTCA	2-10-2MOE	1094
						384545	9243	9254	CAAGTAGGATGT	1-10-1MOE	951
398125	9368	9381	CAGTAAGGAATTTT	2-10-2MOE	913
						398125	9476	9489	CAGTAAGGAATTTT	2-10-2MOE	913
401409	9516	9529	ATTCTTAACACAGA	2-10-2MOE	991
						147096	9594	9605	TTGTTGTTCCCT	1-10-1MOE	1107
147733	9597	9608	TTCTTGATGTCC	1-10-1MOE	891
						147720	9689	9700	GATCTCTCGAGT	1-10-1MOE	1117

147096	9702	9713	TTGTTGTTCCCT	1-10-1MOE	1107
						147733	9705	9716	TTCTTGATGTCC	1-10-1MOE	891
147720	9797	9808	GATCTCTCGAGT	1-10-1MOE	1117
						147746	9963	9974	TAAAAACAACAA	1-10-1MOE	1073
147746	9966	9977	TAAAAACAACAA	1-10-1MOE	1073
						147746	9969	9980	TAAAAACAACAA	1-10-1MOE	1073
147746	9991	10002	TAAAAACAACAA	1-10-1MOE	1073
						147746	10071	10082	TAAAAACAACAA	1-10-1MOE	1073
147746	10074	10085	TAAAAACAACAA	1-10-1MOE	1073
						147746	10077	10088	TAAAAACAACAA	1-10-1MOE	1073
147746	10099	10110	TAAAAACAACAA	1-10-1MOE	1073
						398134	10153	10166	TAGCTTAATGTAAC	2-10-2MOE	1095
147085	10221	10232	TCTACACCAGGT	1-10-1MOE	961
						398134	10261	10274	TAGCTTAATGTAAC	2-10-2MOE	1095
390030	10278	10289	TTTATAAAACTG	1-10-1MOE	1074
						147084	10328	10339	CTACACCAGGTC	1-10-1MOE	993
147711	10684	10695	AAGGGCCCTGGG	1-10-1MOE	1040
						398128	11333	11346	CTAAATTTAGTTCA	2-10-2MOE	911
398128	11340	11353	CTAAATTTAGTTCA	2-10-2MOE	911
						147730	11783	11794	CTTGTCCATCAG	1-10-1MOE	1121
147731	11789	11800	TTTCCTCTTGTC	1-10-1MOE	934
						147730	11790	11801	CTTGTCCATCAG	1-10-1MOE	1121
147731	11796	11807	TTTCCTCTTGTC	1-10-1MOE	934
						147707	11960	11971	TAGTCATTATCT	1-10-1MOE	977
147090	12008	12019	TTCCCTCTACAC	1-10-1MOE	955
						147091	12009	12020	GTTCCCTCTACA	1-10-1MOE	1004
147091	12014	12025	GTTCCCTCTACA	1-10-1MOE	1004
						398096	12141	12154	GGAGAAGCGCAGCT	2-10-2MOE	1015
147735	12143	12154	GGAGAAGCGCAG	1-10-1MOE	1016
						398096	12146	12159	GGAGAAGCGCAGCT	2-10-2MOE	1015
147735	12148	12159	GGAGAAGCGCAG	1-10-1MOE	1016
						398166	12209	12220	GGGCTTCTTCCA	1-10-1MOE	1070
398166	12214	12225	GGGCTTCTTCCA	1-10-1MOE	1070
						398135	12303	12316	GACTACATTTTACA	2-10-2MOE	912
147741	12389	12400	CACCCACTGGTG	1-10-1MOE	1055
						147741	12394	12405	CACCCACTGGTG	1-10-1MOE	1055
398125	12431	12444	CAGTAAGGAATTTT	2-10-2MOE	913
						147714	12585	12596	TTCTGCTCCCAC	1-10-1MOE	986
147718	12594	12605	TAATATGACTTG	1-10-1MOE	998
						398125	12612	12625	CAGTAAGGAATTTT	2-10-2MOE	913
147737	12803	12814	ACAGCCAGGTAG	1-10-1MOE	1067
						147746	12876	12887	TAAAAACAACAA	1-10-1MOE	1073
147691	12900	12911	GAGGTGGGAAAA	1-10-1MOE	966
						398136	12915	12928	TTGTGACATCTAGG	2-10-2MOE	1096
147737	12984	12995	ACAGCCAGGTAG	1-10-1MOE	1067
						147746	13057	13068	TAAAAACAACAA	1-10-1MOE	1073
147691	13081	13092	GAGGTGGGAAAA	1-10-1MOE	966
						398136	13096	13109	TTGTGACATCTAGG	2-10-2MOE	1096
398138	13254	13267	AACATCAAGCTTGA	2-10-2MOE	931
						398138	13435	13448	AACATCAAGCTTGA	2-10-2MOE	931
147691	13488	13499	GAGGTGGGAAAA	1-10-1MOE	966
						147681	13659	13670	ATGTCATTAAAC	1-10-1MOE	965
147691	13669	13680	GAGGTGGGAAAA	1-10-1MOE	966
						389965	13839	13850	CTGCAACATGAT	1-10-1MOE	1018

389764	13839	13850	CTGCAACATGAT	1-9-2MOE	1018
						147681	13840	13851	ATGTCATTAAAC	1-10-1MOE	965
389965	14020	14031	CTGCAACATGAT	1-10-1MOE	1018
						389764	14020	14031	CTGCAACATGAT	1-9-2MOE	1018
389948	14067	14078	CCGTTGGACCCC	1-10-1MOE	915
						147736	14123	14134	AGGTAGGAGAAG	1-10-1MOE	963
389948	14248	14259	CCGTTGGACCCC	1-10-1MOE	915
						147738	14279	14290	TGGGTGGCCGGG	1-10-1MOE	1069
147736	14304	14315	AGGTAGGAGAAG	1-10-1MOE	963
						147731	14411	14422	TTTCCTCTTGTC	1-10-1MOE	934
147738	14461	14472	TGGGTGGCCGGG	1-10-1MOE	1069
						147692	14475	14486	CTCACCTTCATG	1-10-1MOE	1113
147731	14593	14604	TTTCCTCTTGTC	1-10-1MOE	934
						389950	14614	14625	CCCTGAAGGTTC	1-10-1MOE	1063
147692	14657	14668	CTCACCTTCATG	1-10-1MOE	1113
						147717	14750	14761	ATCTTCAGAGAT	1-10-1MOE	996
147698	14754	14765	CCCGCCACCACC	1-10-1MOE	928
						389950	14796	14807	CCCTGAAGGTTC	1-10-1MOE	1063
398112	14863	14876	CAGCCTGGCACCTA	2-10-2MOE	1072
						398121	14875	14888	GTGCCTAGCACAGA	2-10-2MOE	1097
147717	14932	14943	ATCTTCAGAGAT	1-10-1MOE	996
						398112	15045	15058	CAGCCTGGCACCTA	2-10-2MOE	1072
398121	15057	15070	GTGCCTAGCACAGA	2-10-2MOE	1097
						147730	15117	15128	CTTGTCCATCAG	1-10-1MOE	1121
147730	15299	15310	CTTGTCCATCAG	1-10-1MOE	1121
						401407	15339	15352	CAGCTTAGGCAGAG	2-10-2MOE	983
398167	15556	15567	CAGGCCATGTGG	1-10-1MOE	1059
						147736	16444	16455	AGGTAGGAGAAG	1-10-1MOE	963
147746	16510	16521	TAAAAACAACAA	1-10-1MOE	1073
						147738	16590	16601	TGGGTGGCCGGG	1-10-1MOE	1069
147736	16610	16621	AGGTAGGAGAAG	1-10-1MOE	963
						398167	16631	16642	CAGGCCATGTGG	1-10-1MOE	1059
401411	16657	16670	AGCCGCCTGAAGTG	2-10-2MOE	999
						147746	16676	16687	TAAAAACAACAA	1-10-1MOE	1073
398144	16745	16758	GACAGCTTCTATAA	2-10-2MOE	916
						147738	16756	16767	TGGGTGGCCGGG	1-10-1MOE	1069
398167	16797	16808	CAGGCCATGTGG	1-10-1MOE	1059
						398144	16911	16924	GACAGCTTCTATAA	2-10-2MOE	916
389965	17096	17107	CTGCAACATGAT	1-10-1MOE	1018
						389764	17096	17107	CTGCAACATGAT	1-9-2MOE	1018
389965	17264	17275	CTGCAACATGAT	1-10-1MOE	1018
						389764	17264	17275	CTGCAACATGAT	1-9-2MOE	1018
147709	17406	17417	CCATTTTTATCA	1-10-1MOE	978
						147745	17443	17454	TTGACCAGGAAG	1-10-1MOE	1058
147746	17497	17508	TAAAAACAACAA	1-10-1MOE	1073
						147720	17589	17600	GATCTCTCGAGT	1-10-1MOE	1117
147745	17611	17622	TTGACCAGGAAG	1-10-1MOE	1058
						147695	17634	17645	TCATTCCCCACT	1-10-1MOE	984
147746	17665	17676	TAAAAACAACAA	1-10-1MOE	1073
						147088	17707	17718	CCCTCTACACCA	1-10-1MOE	1050
147720	17757	17768	GATCTCTCGAGT	1-10-1MOE	1117
						147711	17808	17819	AAGGGCCCTGGG	1-10-1MOE	1040
147711	17976	17987	AAGGGCCCTGGG	1-10-1MOE	1040
						398139	18049	18062	AGTGACTGACCACA	2-10-2MOE	917

398139	18217	18230	AGTGACTGACCACA	2-10-2MOE	917
						398140	18596	18609	GTAGCATAGAGCCT	2-10-2MOE	918
398140	18764	18777	GTAGCATAGAGCCT	2-10-2MOE	918
						398167	18927	18938	CAGGCCATGTGG	1-10-1MOE	1059
398167	19095	19106	CAGGCCATGTGG	1-10-1MOE	1059
						147724	19147	19158	GAAATTGAGGAA	1-10-1MOE	1139
147746	19207	19218	TAAAAACAACAA	1-10-1MOE	1073
						147724	19315	19326	GAAATTGAGGAA	1-10-1MOE	1139
147740	19348	19359	TGTGAGGCTCCA	1-10-1MOE	1062
						147746	19375	19386	TAAAAACAACAA	1-10-1MOE	1073
147729	19386	19397	GTAAGAGGCAGG	1-10-1MOE	920
						147701	19503	19514	CCATGGCGGGAC	1-10-1MOE	921
147711	19508	19519	AAGGGCCCTGGG	1-10-1MOE	1040
						147740	19516	19527	TGTGAGGCTCCA	1-10-1MOE	1062
147718	19617	19628	TAATATGACTTG	1-10-1MOE	998
						390030	19618	19629	TTTATAAAACTG	1-10-1MOE	1074
147679	19635	19646	CAAAAGGATCCC	1-10-1MOE	907
						147711	19676	19687	AAGGGCCCTGGG	1-10-1MOE	1040
147694	19747	19758	CAGCCTACCAGT	1-10-1MOE	1098
						147718	19785	19796	TAATATGACTTG	1-10-1MOE	998
390030	19786	19797	TTTATAAAACTG	1-10-1MOE	1074
						147679	19803	19814	CAAAAGGATCCC	1-10-1MOE	907
147698	19852	19863	CCCGCCACCACC	1-10-1MOE	928
						147694	19915	19926	CAGCCTACCAGT	1-10-1MOE	1098
147704	20011	20022	TTGTTCTTAGGA	1-10-1MOE	1012
						147698	20020	20031	CCCGCCACCACC	1-10-1MOE	928
398142	20485	20498	CCAGCACACTGGAA	2-10-2MOE	923
						147078	20514	20525	CCTTCCACTGAT	1-10-1MOE	1044
147079	20515	20526	TCCTTCCACTGA	1-10-1MOE	1001
						147080	20516	20527	CTCCTTCCACTG	1-10-1MOE	1021
398143	20561	20574	GTCAGTCCCAGCTA	2-10-2MOE	924
						389965	20620	20631	CTGCAACATGAT	1-10-1MOE	1018
389764	20620	20631	CTGCAACATGAT	1-9-2MOE	1018
						398142	20653	20666	CCAGCACACTGGAA	2-10-2MOE	923
147078	20682	20693	CCTTCCACTGAT	1-10-1MOE	1044
						147079	20683	20694	TCCTTCCACTGA	1-10-1MOE	1001
147080	20684	20695	CTCCTTCCACTG	1-10-1MOE	1021
						147080	20704	20715	CTCCTTCCACTG	1-10-1MOE	1021
147081	20705	20716	GCTCCTTCCACT	1-10-1MOE	1006
						398143	20729	20742	GTCAGTCCCAGCTA	2-10-2MOE	924
389965	20788	20799	CTGCAACATGAT	1-10-1MOE	1018
						389764	20788	20799	CTGCAACATGAT	1-9-2MOE	1018
147746	20870	20881	TAAAAACAACAA	1-10-1MOE	1073
						147080	20872	20883	CTCCTTCCACTG	1-10-1MOE	1021
147081	20873	20884	GCTCCTTCCACT	1-10-1MOE	1006
						147746	21038	21049	TAAAAACAACAA	1-10-1MOE	1073
147717	21080	21091	ATCTTCAGAGAT	1-10-1MOE	996
						147076	21222	21233	TTCCACTGATCC	1-10-1MOE	1029
147076	21390	21401	TTCCACTGATCC	1-10-1MOE	1029
						398094	21441	21454	ATCAGCCAGACAGA	2-10-2MOE	1010
147746	21465	21476	TAAAAACAACAA	1-10-1MOE	1073
						398094	21609	21622	ATCAGCCAGACAGA	2-10-2MOE	1010
398169	21610	21621	TCAGCCAGACAG	1-10-1MOE	909
						147746	21633	21644	TAAAAACAACAA	1-10-1MOE	1073

147738	21884	21895	TGGGTGGCCGGG	1-10-1MOE	1069
						147743	22045	22056	AGGGCTTCCAGT	1-10-1MOE	1042
147738	22052	22063	TGGGTGGCCGGG	1-10-1MOE	1069
						147683	22107	22118	GCTTACGATTGT	1-10-1MOE	922
147743	22213	22224	AGGGCTTCCAGT	1-10-1MOE	1042
						147681	22566	22577	ATGTCATTAAAC	1-10-1MOE	965
389950	22619	22630	CCCTGAAGGTTC	1-10-1MOE	1063
						147681	22734	22745	ATGTCATTAAA C	1-10-1MOE	965
147736	22759	22770	AGGTAGGAGAAG	1-10-1MOE	963
						389950	22787	22798	CCCTGAAGGTTC	1-10-1MOE	1063
389949	22794	22805	GCGCGAGCCCGA	1-10-1MOE	1061
						147736	22927	22938	AGGTAGGAGAAG	1-10-1MOE	963
389949	22962	22973	GCGCGAGCCCGA	1-10-1MOE	1061
						398144	22962	22975	GACAGCTTCTATAA	2-10-2MOE	916
398142	23008	23021	CCAGCACACTGGAA	2-10-2MOE	923
						147727	23019	23030	CAGTGGACCACA	1-10-1MOE	1128
398169	23064	23075	TCAGCCAGACAG	1-10-1MOE	909
						398144	23130	23143	GACAGCTTCTATAA	2-10-2MOE	916
398145	23154	23167	ACATGTCAGTAATT	2-10-2MOE	1099
						398142	23176	23189	CCAGCACACTGGAA	2-10-2MOE	923
147727	23187	23198	CAGTGGACCACA	1-10-1MOE	1128
						147735	23243	23254	GGAGAAGCGCAG	1-10-1MOE	1016
398145	23322	23335	ACATGTCAGTAATT	2-10-2MOE	1099
						147735	23411	23422	GGAGAAGCGCAG	1-10-1MOE	1016
398146	23478	23491	CTCATGGACACAAA	2-10-2MOE	1100
						398146	23646	23659	CTCATGGACACAAA	2-10-2MOE	1100
398147	23784	23797	CTACAGGACAATAC	2-10-2MOE	957
						398114	23853	23866	AGGCATATAGCAGA	2-10-2MOE	1075
398147	23952	23965	CTACAGGACAATAC	2-10-2MOE	957
						398114	24021	24034	AGGCATATAGCAGA	2-10-2MOE	1075
147702	24319	24330	CTGGTAAATAGC	1-10-1MOE	898
						147702	24487	24498	CTGGTAAATAGC	1-10-1MOE	898
389965	24543	24554	CTGCAACATGAT	1-10-1MOE	1018
						389764	24543	24554	CTGCAACATGAT	1-9-2MOE	1018
147713	24602	24613	CTCCCACACCAT	1-10-1MOE	985
						389965	24711	24722	CTGCAACATGAT	1-10-1MOE	1018
389764	24711	24722	CTGCAACATGAT	1-9-2MOE	1018
						147684	24918	24929	ACCCAGTCAGGG	1-10-1MOE	964
147684	25086	25097	ACCCAGTCAGGG	1-10-1MOE	964
						398148	25152	25165	TCATAACTATTAAG	2-10-2MOE	981
398144	25192	25205	GACAGCTTCTATAA	2-10-2MOE	916
						147746	25216	25227	TAAAAACAACAA	1-10-1MOE	1073
147736	25313	25324	AGGTAGGAGAAG	1-10-1MOE	963
						398148	25320	25333	TCATAACTATTAAG	2-10-2MOE	981
398143	25337	25350	GTCAGTCCCAGCTA	2-10-2MOE	924
						398144	25360	25373	GACAGCTTCTATAA	2-10-2MOE	916
147746	25384	25395	TAAAAACAACAA	1-10-1MOE	1073
						147691	25442	25453	GAGGTGGGAAAA	1-10-1MOE	966
147736	25481	25492	AGGTAGGAGAAG	1-10-1MOE	963
						398130	25504	25517	TTAGTATGACAGCT	2-10-2MOE	925
147691	25610	25621	GAGGTGGGAAAA	1-10-1MOE	966
						147721	25662	25673	AATGCAGGATCT	1-10-1MOE	1118
398130	25672	25685	TTAGTATGACAGCT	2-10-2MOE	925
						147688	25750	25761	TCCCAAACAAAT	1-10-1MOE	990

147746	25810	25821	TAAAAACAACAA	1-10-1MOE	1073
						147721	25830	25841	AATGCAGGATCT	1-10-1MOE	1118
147688	25918	25929	TCCCAAACAAAT	1-10-1MOE	990
						147746	25978	25989	TAAAAACAACAA	1-10-1MOE	1073
147746	26172	26183	TAAAAACAACAA	1-10-1MOE	1073
						147746	26340	26351	TAAAAACAACAA	1-10-1MOE	1073
398149	26492	26505	GGAAGTTTTCAAGT	2-10-2MOE	1101
						398150	26526	26539	GAATCTGGAGGTAA	2-10-2MOE	1102
398149	26641	26654	GGAAGTTTTCAAGT	2-10-2MOE	1101
						398150	26675	26688	GAATCTGGAGGTAA	2-10-2MOE	1102
147729	26712	26723	GTAAGAGGCAGG	1-10-1MOE	920
						398151	26718	26731	TCAGTGTAGGAAGA	2-10-2MOE	926
147729	26861	26872	GTAAGAGGCAGG	1-10-1MOE	920
						398151	26867	26880	TCAGTGTAGGAAGA	2-10-2MOE	926
147728	26917	26928	GCCAGACAGAAG	1-10-1MOE	1013
						147728	27066	27077	GCCAGACAGAAG	1-10-1MOE	1013
147076	27258	27269	TTCCACTGATCC	1-10-1MOE	1029
						147731	27267	27278	TTTCCTCTTGTC	1-10-1MOE	934
147076	27407	27418	TTCCACTGATCC	1-10-1MOE	1029
						147731	27416	27427	TTTCCTCTTGTC	1-10-1MOE	934
398152	27559	27572	TGAATATACAGATG	2-10-2MOE	927
						398152	27708	27721	TGAATATACAGATG	2-10-2MOE	927
147696	28265	28276	TGGATGATTGGC	1-10-1MOE	906
						147696	28414	28425	TGGATGATTGGC	1-10-1MOE	906
147698	28481	28492	CCCGCCACCACC	1-10-1MOE	928
						147720	28662	28673	GATCTCTCGAGT	1-10-1MOE	1117
389965	28714	28725	CTGCAACATGAT	1-10-1MOE	1018
						389764	28714	28725	CTGCAACATGAT	1-9-2MOE	1018
389965	28861	28872	CTGCAACATGAT	1-10-1MOE	1018
						389764	28861	28872	CTGCAACATGAT	1-9-2MOE	1018
398153	28980	28993	ATTTCTCTTACAGG	2-10-2MOE	948
						398153	29126	29139	ATTTCTCTTACAGG	2-10-2MOE	948
147719	29570	29581	CCAACTCCAACT	1-10-1MOE	1116
						398154	29692	29705	AGCCCCTTGGCCGT	2-10-2MOE	1103
147719	29715	29726	CCAACTCCAACT	1-10-1MOE	1116
						398155	29785	29798	TGTTTTTACACAGA	2-10-2MOE	970
398154	29837	29850	A GCCCCTTGGCCGT	2-10-2MOE	1103
						401384	29905	29918	TGAACACATCACTA	2-10-2MOE	933
398155	29930	29943	TGTTTTTACACAGA	2-10-2MOE	970
						390030	29945	29956	TTTATAAAACTG	1-10-1MOE	1074
390030	30090	30101	TTTATAAAACTG	1-10-1MOE	1074
						398156	30141	30154	GAATACTTCAAATC	2-10-2MOE	1104
398156	30286	30299	GAATACTTCAAATC	2-10-2MOE	1104
						389948	30384	30395	CCGTTGGACCCC	1-10-1MOE	915
389948	30530	30541	CCGTTGGACCCC	1-10-1MOE	915
						398142	30591	30604	CCAGCACACTGGAA	2-10-2MOE	923
147744	30654	30665	AGGAAGGGCTTC	1-10-1MOE	1043
						147093	30689	30700	TTGTTCCCTCTA	1-10-1MOE	929
398142	30738	30751	CCAGCACACTGGAA	2-10-2MOE	923
						147744	30801	30812	AGGAAGGGCTTC	1-10-1MOE	1043
398168	31082	31093	TCGGACTTTGAA	1-10-1MOE	1008
						147746	31105	31116	TAAAAACAACAA	1-10-1MOE	1073
398168	31230	31241	TCGGACTTTGAA	1-10-1MOE	1008
						390030	31329	31340	TTTATAAAACTG	1-10-1MOE	1074

147736	31458	31469	AGGTAGGAGAAG	1-10-1MOE	963
						390030	31477	31488	TTTATAAAACTG	1-10-1MOE	1074
147736	31606	31617	AGGTAGGAGAAG	1-10-1MOE	963
						147698	31713	31724	CCCGCCACCACC	1-10-1MOE	928
384545	31829	31840	CAAGTAGGATGT	1-10-1MOE	951
						147698	31861	31872	CCCGCCACCACC	1-10-1MOE	928
147723	31941	31952	GACTCCAAAGTC	1-10-1MOE	892
						384545	31977	31988	CAAGTAGGATGT	1-10-1MOE	951
147692	32061	32072	CTCACCTTCATG	1-10-1MOE	1113
						147723	32089	32100	GACTCCAAAGTC	1-10-1MOE	892
147692	32209	32220	CTCACCTTCATG	1-10-1MOE	1113
						147089	32535	32546	TCCCTCTACACC	1-10-1MOE	956
401396	32569	32582	TGCAGGATGTTGAG	2-10-2MOE	945
						147730	32714	32725	CTTGTCCATCAG	1-10-1MOE	1121
398165	32854	32865	GTTCTTAGGAAG	1-10-1MOE	968
						147730	32862	32873	CTTGTCCATCAG	1-10-1MOE	1121
389950	32949	32960	CCCTGAAGGTTC	1-10-1MOE	1063
						398165	33002	33013	GTTCTTAGGAAG	1-10-1MOE	968
147736	33012	33023	AGGTAGGAGAAG	1-10-1MOE	963
						368352	33056	33069	CTGATCCTGCACTG	2-10-2MOE	1105
147081	33073	33084	GCTCCTTCCACT	1-10-1MOE	1006
						368360	33073	33086	AAGCTCCTTCCACT	2-10-2MOE	1035
147082	33074	33085	AGCTCCTTCCA C	1-10-1MOE	1036
						389950	33097	33108	CCCTGAAGGTTC	1-10-1MOE	1063
147736	33160	33171	AGGTAGGAGAAG	1-10-1MOE	963
						368352	33204	33217	CTGATCCTGCACTG	2-10-2MOE	1105
147081	33221	33232	GCTCCTTCCACT	1-10-1MOE	1006
						147082	33222	33233	AGCTCCTTCCAC	1-10-1MOE	1036
398138	33244	33257	AACATCAAGCTTGA	2-10-2MOE	931
						147746	33250	33261	TAAAAACAACAA	1-10-1MOE	1073
398138	33392	33405	AACATCAAGCTTGA	2-10-2MOE	931
						147746	33398	33409	TAAAAACAACAA	1-10-1MOE	1073
147732	33652	33663	GGGTCTTTCCTC	1-10-1MOE	1122
						147724	33733	33744	GAAATTGAGGAA	1-10-1MOE	1139
147732	33800	33811	GGGTCTTTCCTC	1-10-1MOE	1122
						147724	33881	33892	GAAATTGAGGAA	1-10-1MOE	1139
147719	33976	33987	CCAACTCCAACT	1-10-1MOE	1116
						147746	34034	34045	TAAAAACAACAA	1-10-1MOE	1073
398129	34045	34058	TTTGAGGAGCTATT	2-10-2MOE	1106
						147719	34124	34135	CCAACTCCAACT	1-10-1MOE	1116
147721	34156	34167	AATGCAGGATCT	1-10-1MOE	1118
						398129	34193	34206	TTTGAGGAGCTATT	2-10-2MOE	1106
147721	34304	34315	AATGCAGGATCT	1-10-1MOE	1118
						147746	34606	34617	TAAAAACAACAA	1-10-1MOE	1073
398165	34704	34715	GTTCTTAGGAAG	1-10-1MOE	968
						147746	34754	34765	TAAAAACAACAA	1-10-1MOE	1073
398165	34852	34863	GTTCTTAGGAAG	1-10-1MOE	968
						147717	34893	34904	ATCTTCAGAGAT	1-10-1MOE	996
147719	34976	34987	CCAACTCCAACT	1-10-1MOE	1116
						147092	34987	34998	TGTTCCCTCTAC	1-10-1MOE	901
147719	35124	35135	CCAACTCCAACT	1-10-1MOE	1116
						147092	35135	35146	TGTTCCCTCTAC	1-10-1MOE	901
147736	35248	35259	AGGTAGGAGAAG	1-10-1MOE	963
						147738	35391	35402	TGGGTGGCCGGG	1-10-1MOE	1069

147736	35396	35407	AGGTAGGAGAAG	1-10-1MOE	963
						147738	35539	35550	TGGGTGGCCGGG	1-10-1MOE	1069
147691	35554	35565	GAGGTGGGAAAA	1-10-1MOE	966
						147691	35702	35713	GAGGTGGGAAAA	1-10-1MOE	966
147746	35814	35825	TAAAAACAACAA	1-10-1MOE	1073
						147733	35889	35900	TTCTTGATGTCC	1-10-1MOE	891
147733	35923	35934	TTCTTGATGTCC	1-10-1MOE	891
						147746	35962	35973	TAAAAACAACAA	1-10-1MOE	1073
147726	35978	35989	TGACTCTCGGAC	1-10-1MOE	1120
						147733	36037	36048	TTCTTGATGTCC	1-10-1MOE	891
147733	36071	36082	TTCTTGATGTCC	1-10-1MOE	891
						147726	36126	36137	TGACTCTCGGAC	1-10-1MOE	1120
147736	36359	36370	AGGTAGGAGAAG	1-10-1MOE	963
						147691	36360	36371	GAGGTGGGAAAA	1-10-1MOE	966
147736	36507	36518	AGGTAGGAGAAG	1-10-1MOE	963
						147691	36508	36519	GAGGTGGGAAAA	1-10-1MOE	966
147746	36564	36575	TAAAAACAACAA	1-10-1MOE	1073
						147723	36575	36586	GACTCCAAAGTC	1-10-1MOE	892
147731	36620	36631	TTTCCTCTTGTC	1-10-1MOE	934
						147723	36723	36734	GACTCCAAAGTC	1-10-1MOE	892
147731	36768	36779	TTTCCTCTTGTC	1-10-1MOE	934
						398169	37174	37185	TCAGCCAGACAG	1-10-1MOE	909
147688	37380	37391	TCCCAAACAAAT	1-10-1MOE	990
						147688	37528	37539	TCCCAAACAAAT	1-10-1MOE	990
147714	37881	37892	TTCTGCTCCCAC	1-10-1MOE	986
						147714	38029	38040	TTCTGCTCCCAC	1-10-1MOE	986
147681	38364	38375	ATGTCATTAAAC	1-10-1MOE	965
						147736	38766	38777	AGGTAGGAGAAG	1-10-1MOE	963
147738	38909	38920	TGGGTGGCCGGG	1-10-1MOE	1069
						147736	38914	38925	AGGTAGGAGAAG	1-10-1MOE	963
147738	39057	39068	TGGGTGGCCGGG	1-10-1MOE	1069
						390030	39249	39260	TTTATAAAACTG	1-10-1MOE	1074
390030	39397	39408	TTTATAAAACTG	1-10-1MOE	1074
						147717	39545	39556	ATCTTCAGAGAT	1-10-1MOE	996
147717	39693	39704	ATCTTCAGAGAT	1-10-1MOE	996
						147746	39729	39740	TAAAAACAACAA	1-10-1MOE	1073
147746	39789	39800	TAAAAACAACAA	1-10-1MOE	1073
						147691	39829	39840	GAGGTGGGAAAA	1-10-1MOE	966
147746	39877	39888	TAAAAACAACAA	1-10-1MOE	1073
						147691	39977	39988	GAGGTGGGAAAA	1-10-1MOE	966
147727	39983	39994	CAGTGGACCACA	1-10-1MOE	1128
						147727	40131	40142	CAGTGGACCACA	1-10-1MOE	1128
147746	40333	40344	TAAAAACAACAA	1-10-1MOE	1073
						147719	40457	40468	CCAACTCCAACT	1-10-1MOE	1116
147679	40467	40478	CAAAAGGATCCC	1-10-1MOE	907
						147746	40478	40489	TAAAAACAACAA	1-10-1MOE	1073
147741	40565	40576	CACCCACTGGTG	1-10-1MOE	1055
						398166	40589	40600	GGGCTTCTTCCA	1-10-1MOE	1070
147719	40605	40616	CCAACTCCAACT	1-10-1MOE	1116
						147679	40615	40626	CAAAAGGATCCC	1-10-1MOE	907
147746	40626	40637	TAAAAACAACAA	1-10-1MOE	1073
						147735	40662	40673	GGAGAAGCGCAG	1-10-1MOE	1016
147746	40706	40717	TAAAAACAACAA	1-10-1MOE	1073
						147741	40713	40724	CACCCACTGGTG	1-10-1MOE	1055

398166	40737	40748	GGGCTTCTTCCA	1-10-1MOE	1070
						147735	40810	40821	GGAGAAGCGCAG	1-10-1MOE	1016
147746	40854	40865	TAAAAACAACAA	1-10-1MOE	1073
						147718	41218	41229	TAATATGACTTG	1-10-1MOE	998
147717	41221	41232	ATCTTCAGAGAT	1-10-1MOE	996
						147717	41369	41380	ATCTTCAGAGAT	1-10-1MOE	996
147723	41627	41638	GACTCCAAAGTC	1-10-1MOE	892
						147717	41747	41758	ATCTTCAGAGAT	1-10-1MOE	996
147723	41775	41786	GACTCCAAAGTC	1-10-1MOE	892
						390030	41908	41919	TTTATAAAACTG	1-10-1MOE	1074
390030	42056	42067	TTTATAAAACTG	1-10-1MOE	1074
						398153	42157	42170	ATTTCTCTTACAGG	2-10-2MOE	948
398153	42305	42318	ATTTCTCTTACAGG	2-10-2MOE	948
						147690	42423	42434	TGAAGTTAATTC	1-10-1MOE	1138
147695	42521	42532	TCATTCCCCACT	1-10-1MOE	984
						147710	42543	42554	TATAGCTCCTCT	1-10-1MOE	994
147690	42571	42582	TGAAGTTAATTC	1-10-1MOE	1138
						147695	42669	42680	TCATTCCCCACT	1-10-1MOE	984
147078	43321	43332	CCTTCCACTGAT	1-10-1MOE	1044
						147079	43322	43333	TCCTTCCACTGA	1-10-1MOE	1001
147716	43329	43340	TTAACGAGCCTT	1-10-1MOE	949
						147078	43469	43480	CCTTCCACTGAT	1-10-1MOE	1044
147079	43470	43481	TCCTTCCACTGA	1-10-1MOE	1001
						147080	43471	43482	CTCCTTCCACTG	1-10-1MOE	1021
398102	43837	43850	CTACCTGAGGATTT	2-10-2MOE	899
						147074	43848	43859	CCACTGATCCTG	1-10-1MOE	845
401408	43871	43884	CAATGAAGCACAGG	2-10-2MOE	989
						398102	43985	43998	CTACCTGAGGATTT	2-10-2MOE	899
147736	44137	44148	AGGTAGGAGAAG	1-10-1MOE	963
						147746	44140	44151	TAAAAACAACAA	1-10-1MOE	1073
147687	44206	44217	CGACACGGGAAC	1-10-1MOE	950
						147743	44223	44234	AGGGCTTCCAGT	1-10-1MOE	1042
384545	44242	44253	CAAGTAGGATGT	1-10-1MOE	951
						147736	44285	44296	AGGTAGGAGAAG	1-10-1MOE	963
147743	44371	44382	AGGGCTTCCAGT	1-10-1MOE	1042
						384545	44390	44401	CAAGTAGGATGT	1-10-1MOE	951
147728	44589	44600	GCCAGACAGAAG	1-10-1MOE	1013
						389948	44628	44639	CCGTTGGACCCC	1-10-1MOE	915
147720	44703	44714	GATCTCTCGAGT	1-10-1MOE	1117
						147728	44729	44740	GCCAGACAGAAG	1-10-1MOE	1013
147728	44737	44748	GCCAGACAGAAG	1-10-1MOE	1013
						389948	44776	44787	CCGTTGGACCCC	1-10-1MOE	915
147720	44851	44862	GATCTCTCGAGT	1-10-1MOE	1117
						398110	44861	44874	GTTCCCTTTGCAGG	2-10-2MOE	952
147728	44877	44888	GCCAGACAGAAG	1-10-1MOE	1013
						147705	45092	45103	CGGTTTTTGTTC	1-10-1MOE	1002
147705	45240	45251	CGGTTTTTGTTC	1-10-1MOE	1002
						147681	45337	45348	ATGTCATTAAAC	1-10-1MOE	965
147681	45485	45496	ATGTCATTAAAC	1-10-1MOE	965
						147096	45660	45671	TTGTTGTTCCCT	1-10-1MOE	1107
147096	45808	45819	TTGTTGTTCCCT	1-10-1MOE	1107
						368368	45976	45989	TCCACTGATCCTTA	2-10-2MOE	1127
147074	45977	45988	CCACTGATCCTG	1-10-1MOE	845
						147075	45978	45989	TCCACTGATCCT	1-10-1MOE	1026

147076	45979	45990	TTCCACTGATCC	1-10-1MOE	1029
						368368	46124	46137	TCCACTGATCCTTA	2-10-2MOE	1127
147075	46126	46137	TCCACTGATCCT	1-10-1MOE	1026
						147076	46127	46138	TTCCACTGATCC	1-10-1MOE	1029
147705	46555	46566	CGGTTTTTGTTC	1-10-1MOE	1002
						147714	46685	46696	TTCTGCTCCCAC	1-10-1MOE	986
147705	46703	46714	CGGTTTTTGTTC	1-10-1MOE	1002
						147714	46833	46844	TTCTGCTCCCAC	1-10-1MOE	986
390030	47007	47018	TTTATAAAACTG	1-10-1MOE	1074
						147746	47023	47034	TAAAAACAACAA	1-10-1MOE	1073
147746	47171	47182	TAAAAACAACAA	1-10-1MOE	1073
						147085	47607	47618	TCTACACCAGGT	1-10-1MOE	961
147746	47609	47620	TAAAAACAACAA	1-10-1MOE	1073
						147089	47611	47622	TCCCTCTACACC	1-10-1MOE	956
147091	47613	47624	GTTCCCTCTACA	1-10-1MOE	1004
						401384	47689	47702	TGAACACATCACTA	2-10-2MOE	933
147691	47729	47740	GAGGTGGGAAAA	1-10-1MOE	966
						147085	47755	47766	TCTACACCAGGT	1-10-1MOE	961
147087	47757	47768	CCTCTACACCAG	1-10-1MOE	982
						147090	47760	47771	TTCCCTCTACAC	1-10-1MOE	955
147091	47761	47772	GTTCCCTCTACA	1-10-1MOE	1004
						147099	47770	47781	GAGTTGTTGTTC	1-10-1MOE	1108
147100	47771	47782	CGAGTTGTTGTT	1-10-1MOE	1109
						390030	47847	47858	TTTATAAAACTG	1-10-1MOE	1074
147691	47877	47888	GAGGTGGGAAAA	1-10-1MOE	966
						147099	47918	47929	GAGTTGTTGTTC	1-10-1MOE	1108
147100	47919	47930	CGAGTTGTTGTT	1-10-1MOE	1109
						390030	47995	48006	TTTATAAAACTG	1-10-1MOE	1074
147074	48222	48233	CCACTGATCCTG	1-10-1MOE	845
						147731	48340	48351	TTTCCTCTTGTC	1-10-1MOE	934
147691	48393	48404	GAGGTGGGAAAA	1-10-1MOE	966
						147731	48488	48499	TTTCCTCTTGTC	1-10-1MOE	934
147691	48541	48552	GAGGTGGGAAAA	1-10-1MOE	966
						398147	48887	48900	CTACAGGACAATAC	2-10-2MOE	957
398147	49035	49048	CTACAGGACAATAC	2-10-2MOE	957
						147074	49525	49536	CCACTGATCCTG	1-10-1MOE	845
398168	49742	49753	TCGGACTTTGAA	1-10-1MOE	1008
						384545	49858	49869	CAAGTAGGATGT	1-10-1MOE	951
398168	49890	49901	TCGGACTTTGAA	1-10-1MOE	1008
						147724	49974	49985	GAAATTGAGGAA	1-10-1MOE	1139
384545	50006	50017	CAAGTAGGATGT	1-10-1MOE	951
						147689	50084	50095	CAGAGAAGGTCT	1-10-1MOE	987
147687	50102	50113	CGACACGGGAAC	1-10-1MOE	950
						147724	50122	50133	GAAATTGAGGAA	1-10-1MOE	1139
147687	50250	50261	CGACACGGGAAC	1-10-1MOE	950
						398117	50389	50402	TTTCCACTTGGGTG	2-10-2MOE	960
147736	50436	50447	AGGTAGGAGAAG	1-10-1MOE	963
						147736	50582	50593	AGGTAGGAGAAG	1-10-1MOE	963
398168	50703	50714	TCGGACTTTGAA	1-10-1MOE	1008
						401397	50822	50835	CTGGTCAGCATTGA	2-10-2MOE	946
147746	51019	51030	TAAAAACAACAA	1-10-1MOE	1073
						147708	51101	51112	TTGATATAGTCA	1-10-1MOE	997
147746	51165	51176	TAAAAACAACAA	1-10-1MOE	1073
						147746	51185	51196	TAAAAACAACAA	1-10-1MOE	1073

147708	51247	51258	TTGATATAGTCA	1-10-1MOE	997
						147081	51287	51298	GCTCCTTCCACT	1-10-1MOE	1006
147082	51288	51299	AGCTCCTTCCAC	1-10-1MOE	1036
						147746	51324	51335	TAAAAACAACAA	1-10-1MOE	1073
147746	51331	51342	TAAAAACAACAA	1-10-1MOE	1073
						147728	51376	51387	GCCAGACAGAAG	1-10-1MOE	1013
147729	51406	51417	GTAAGAGGCAGG	1-10-1MOE	920
						147081	51433	51444	GCTCCTTCCACT	1-10-1MOE	1006
147082	51434	51445	AGCTCCTTCCAC	1-10-1MOE	1036
						147728	51492	51503	GCCAGACAGAAG	1-10-1MOE	1013
147728	51522	51533	GCCAGACAGAAG	1-10-1MOE	1013
						147729	51552	51563	GTAAGAGGCAGG	1-10-1MOE	920
368360	51633	51646	AAGCTCCTTCCACT	2-10-2MOE	1035
						147082	51634	51645	AGCTCCTTCCAC	1-10-1MOE	1036
368361	51635	51648	GAAAGCTCCTTCCA	2-10-2MOE	962
						147728	51638	51649	GCCAGACAGAAG	1-10-1MOE	1013
147695	51644	51655	TCATTCCCCACT	1-10-1MOE	984
						147736	51713	51724	AGGTAGGAGAAG	1-10-1MOE	963
147684	51721	51732	ACCCAGTCAGGG	1-10-1MOE	964
						147081	51779	51790	GCTCCTTCCACT	1-10-1MOE	1006
368360	51779	51792	AAGCTCCTTCCACT	2-10-2MOE	1035
						147082	51780	51791	AGCTCCTTCCAC	1-10-1MOE	1036
368361	51781	51794	GAAAGCTCCTTCCA	2-10-2MOE	962
						147695	51790	51801	TCATTCCCCACT	1-10-1MOE	984
147736	51859	51870	AGGTAGGAGAAG	1-10-1MOE	963
						147077	51988	51999	CTTCCACTGATC	1-10-1MOE	1047
147079	51990	52001	TCCTTCCACTGA	1-10-1MOE	1001
						147746	52064	52075	TAAAAACAACAA	1-10-1MOE	1073
147681	52085	52096	ATGTCATTAAAC	1-10-1MOE	965
						147077	52134	52145	CTTCCACTGATC	1-10-1MOE	1047
147079	52136	52147	TCCTTCCACTGA	1-10-1MOE	1001
						147691	52166	52177	GAGGTGGGAAAA	1-10-1MOE	966
147719	52252	52263	CCAACTCCAACT	1-10-1MOE	1116
						147691	52312	52323	GA GGTGGGAAAA	1-10-1MOE	966
147719	52398	52409	CCAACTCCAACT	1-10-1MOE	1116
						147728	52428	52439	GCCAGACAGAAG	1-10-1MOE	1013
147729	52483	52494	GTAAGAGGCAGG	1-10-1MOE	920
						398167	52527	52538	CAGGCCATGTGG	1-10-1MOE	1059
147682	52571	52582	CGGGTACTATGG	1-10-1MOE	992
						147728	52574	52585	GCCAGACAGAAG	1-10-1MOE	1013
147724	52615	52626	GAAATTGAGGAA	1-10-1MOE	1139
						147729	52629	52640	GTAAGAGGCAGG	1-10-1MOE	920
147703	52670	52681	TGGCTTCATGTC	1-10-1MOE	971
						398167	52673	52684	CAGGCCATGTGG	1-10-1MOE	1059
398165	52708	52719	GTTCTTAGGAAG	1-10-1MOE	968
						147704	52710	52721	TTGTTCTTAGGA	1-10-1MOE	1012
147705	52716	52727	CGGTTTTTGTTC	1-10-1MOE	1002
						147724	52761	52772	GAAATTGAGGAA	1-10-1MOE	1139
398167	52762	52773	CAGGCCATGTGG	1-10-1MOE	1059
						147703	52816	52827	TGGCTTCATGTC	1-10-1MOE	971
398165	52854	52865	GTTCTTAGGAAG	1-10-1MOE	968
						147704	52856	52867	TTGTTCTTAGGA	1-10-1MOE	1012
147705	52862	52873	CGGTTTTTGTTC	1-10-1MOE	1002
						398167	52908	52919	CAGGCCATGTGG	1-10-1MOE	1059

147689	53063	53074	CAGAGAAGGTCT	1-10-1MOE	987
						147727	53111	53122	CAGTGGACCACA	1-10-1MOE	1128
147727	53158	53169	CAGTGGACCACA	1-10-1MOE	1128
						147689	53209	53220	CAGAGAAGGTCT	1-10-1MOE	987
147727	53257	53268	CAGTGGACCACA	1-10-1MOE	1128
						147727	53304	53315	CAGTGGACCACA	1-10-1MOE	1128
147680	53638	53649	GTATGCACTGCT	1-10-1MOE	988
						147722	53650	53661	AAAGTCAGGCCA	1-10-1MOE	1130
147083	53703	53714	TACACCAGGTCA	1-10-1MOE	973
						147085	53705	53716	TCTACACCAGGT	1-10-1MOE	961
147086	53706	53717	CTCTACACCAGG	1-10-1MOE	969
						398167	53724	53735	CAGGCCATGTGG	1-10-1MOE	1059
147684	53747	53758	ACCCAGTCAGGG	1-10-1MOE	964
						147680	53784	53795	GTATGCACTGCT	1-10-1MOE	988
147722	53796	53807	AAAGTCAGGCCA	1-10-1MOE	1130
						147085	53851	53862	TCTACACCAGGT	1-10-1MOE	961
398167	53870	53881	CAGGCCATGTGG	1-10-1MOE	1059
						147684	53893	53904	ACCCAGTCAGGG	1-10-1MOE	964
398155	54026	54039	TGTTTTTACACAGA	2-10-2MOE	970
						147703	54137	54148	TGGCTTCATGTC	1-10-1MOE	971
398155	54172	54185	TGTTTTTACACAGA	2-10-2MOE	970
						147705	54275	54286	CGGTTTTTGTTC	1-10-1MOE	1002
147703	54283	54294	TGGCTTCATGTC	1-10-1MOE	971
						147705	54421	54432	CGGTTTTTGTTC	1-10-1MOE	1002
147727	54853	54864	CAGTGGACCACA	1-10-1MOE	1128
						398165	54963	54974	GTTCTTAGGAAG	1-10-1MOE	968
398090	54963	54976	TTGTTCTTAGGAAG	2-10-2MOE	972
						147704	54965	54976	TTGTTCTTAGGA	1-10-1MOE	1012
147705	54971	54982	CGGTTTTTGTTC	1-10-1MOE	1002
						147727	54999	55010	CAGTGGACCACA	1-10-1MOE	1128
398165	55109	55120	GTTCTTAGGAAG	1-10-1MOE	968
						147704	55111	55122	TTGTTCTTAGGA	1-10-1MOE	1012
147705	55117	55128	CGGTTTTTGTTC	1-10-1MOE	1002
						147083	55352	55363	TACACCAGGTCA	1-10-1MOE	973
147705	55378	55389	CGGTTTTTGTTC	1-10-1MOE	1002
						147705	55524	55535	CGGTTTTTGTTC	1-10-1MOE	1002
147712	55819	55830	ACACCATCTCCC	1-10-1MOE	1005
						147712	55965	55976	ACACCATCTCCC	1-10-1MOE	1005
147733	56289	56300	TTCTTGATGTCC	1-10-1MOE	891
						147707	56300	56311	TAGTCATTATCT	1-10-1MOE	977
147708	56306	56317	TTGATATAGTCA	1-10-1MOE	997
						390030	56321	56332	TTTATAAAACTG	1-10-1MOE	1074
147081	56333	56344	GCTCCTTCCACT	1-10-1MOE	1006
						398166	56335	56346	GGGCTTCTTCCA	1-10-1MOE	1070
147733	56435	56446	TTCTTGATGTCC	1-10-1MOE	891
						147707	56446	56457	TAGTCATTATCT	1-10-1MOE	977
147708	56452	56463	TTGATATAGTCA	1-10-1MOE	997
						390030	56467	56478	TTTATAAAACTG	1-10-1MOE	1074
147081	56479	56490	GCTCCTTCCACT	1-10-1MOE	1006
						398091	56479	56492	GGGCTTCTTCCATT	2-10-2MOE	979
398166	56481	56492	GGGCTTCTTCCA	1-10-1MOE	1070
						368366	56518	56531	CTGATCCTTAGAAG	2-10-2MOE	1019
147743	57612	57623	AGGGCTTCCAGT	1-10-1MOE	1042
						147700	57709	57720	GCGCTAGGCCGC	1-10-1MOE	1110

147743	57758	57769	AGGGCTTCCAGT	1-10-1MOE	1042
						147700	57855	57866	GCGCTAGGCCGC	1-10-1MOE	1110
398093	57963	57976	TCGGACTTTGAAAA	2-10-2MOE	1009
						398168	57965	57976	TCGGACTTTGAA	1-10-1MOE	1008
147698	58105	58116	CCCGCCACCACC	1-10-1MOE	928
						398093	58109	58122	TCGGACTTTGAAAA	2-10-2MOE	1009
398168	58111	58122	TCGGACTTTGAA	1-10-1MOE	1008
						147698	58251	58262	CCCGCCACCACC	1-10-1MOE	928
147735	58279	58290	GGAGAAGCGCAG	1-10-1MOE	1016
						147735	58425	58436	GGAGAAGCGCAG	1-10-1MOE	1016
404135	58946	58959	CATTTCCATGGCCA	2-10-2MOE	1056
						390030	59326	59337	TTTATAAAACTG	1-10-1MOE	1074
147711	59357	59368	AAGGGCCCTGGG	1-10-1MOE	1040
						147743	59382	59393	AGGGCTTCCAGT	1-10-1MOE	1042
147711	59503	59514	AAGGGCCCTGGG	1-10-1MOE	1040
						147743	59528	59539	AGGGCTTCCAGT	1-10-1MOE	1042
147695	59576	59587	TCATTCCCCACT	1-10-1MOE	984
						147713	59716	59727	CTCCCACACCAT	1-10-1MOE	985
147714	59721	59732	TTCTGCTCCCAC	1-10-1MOE	986
						147715	59746	59757	GTTGAGCATGAC	1-10-1MOE	1077
147716	59771	59782	TTAACGAGCCTT	1-10-1MOE	949
						147712	59857	59868	ACACCATCTCCC	1-10-1MOE	1005
147714	59867	59878	TTCTGCTCCCAC	1-10-1MOE	986
						147715	59892	59903	GTTGAGCATGAC	1-10-1MOE	1077
147716	59917	59928	TTAA CGAGCCTT	1-10-1MOE	949
						390030	59993	60004	TTTATAAAACTG	1-10-1MOE	1074
147690	60270	60281	TGAAGTTAATTC	1-10-1MOE	1138
						389949	60325	60336	GCGCGAGCCCGA	1-10-1MOE	1061
147690	60416	60427	TGAAGTTAATTC	1-10-1MOE	1138
						389949	60471	60482	GCGCGAGCCCGA	1-10-1MOE	1061
147746	60619	60630	TAAAAACAACAA	1-10-1MOE	1073
						384545	60676	60687	CAAGTAGGATGT	1-10-1MOE	951
147746	60765	60776	TAAAAACAACAA	1-10-1MOE	1073
						384545	60822	60833	CAAGTAGGATGT	1-10-1MOE	951
147689	60967	60978	CAGAGAAGGTCT	1-10-1MOE	987
						147689	61008	61019	CAGAGAAGGTCT	1-10-1MOE	987
147689	61049	61060	CAGAGAAGGTCT	1-10-1MOE	987
						398105	61121	61134	TGCACAGGCAGGTT	2-10-2MOE	1066
147689	61154	61165	CAGAGAAGGTCT	1-10-1MOE	987
						147689	61195	61206	CAGAGAAGGTCT	1-10-1MOE	987
398105	61267	61280	TGCACAGGCAGGTT	2-10-2MOE	1066
						147692	61365	61376	CTCACCTTCATG	1-10-1MOE	1113
147692	61511	61522	CTCACCTTCATG	1-10-1MOE	1113
						147680	61619	61630	GTATGCACTGCT	1-10-1MOE	988
147078	61755	61766	CCTTCCACTGAT	1-10-1MOE	1044
						147079	61756	61767	TCCTTCCACTGA	1-10-1MOE	1001
147080	61757	61768	CTCCTTCCACTG	1-10-1MOE	1021
						147078	61901	61912	CCTTCCACTGAT	1-10-1MOE	1044
147079	61902	61913	TCCTTCCACTGA	1-10-1MOE	1001
						147080	61903	61914	CTCCTTCCACTG	1-10-1MOE	1021
147088	62361	62372	CCCTCTACACCA	1-10-1MOE	1050
						401384	62573	62586	TGAACACATCACTA	2-10-2MOE	933
147688	62697	62708	TCCCAAACAAAT	1-10-1MOE	990
						147746	63102	63113	TAAAAACAACAA	1-10-1MOE	1073

147721	63225	63236	AATGCAGGATCT	1-10-1MOE	1118
						147742	63226	63237	AACTTCAGTGTC	1-10-1MOE	1041
147746	63248	63259	TAAAAACAACAA	1-10-1MOE	1073
						147682	63337	63348	CGGGTACTATGG	1-10-1MOE	992
147721	63371	63382	AATGCAGGATCT	1-10-1MOE	1118
						147742	63372	63383	AACTTCAGTGTC	1-10-1MOE	1041
147688	63401	63412	TCCCAAACAAAT	1-10-1MOE	990
						147097	63449	63460	GTTGTTGTTCCC	1-10-1MOE	1111
147098	63450	63461	AGTTGTTGTTCC	1-10-1MOE	1112
						401409	63458	63471	ATTCTTAACACAGA	2-10-2MOE	991
147084	63531	63542	CTACACCAGGTC	1-10-1MOE	993
						147688	63547	63558	TCCCAAACAAAT	1-10-1MOE	990
147097	63595	63606	GTTGTTGTTCCC	1-10-1MOE	1111
						147098	63596	63607	AGTTGTTGTTCC	1-10-1MOE	1112
147721	64086	64097	AATGCAGGATCT	1-10-1MOE	1118
						147721	64232	64243	AATGCAGGATCT	1-10-1MOE	1118
147692	64233	64244	CTCACCTTCATG	1-10-1MOE	1113
						147692	64379	64390	CTCACCTTCATG	1-10-1MOE	1113
147729	64633	64644	GTAAGAGGCAGG	1-10-1MOE	920
						401403	64746	64759	TTTCCTAGGAGGTG	2-10-2MOE	967
147729	64779	64790	GTAAGAGGCAGG	1-10-1MOE	920
						147746	65151	65162	TAAAAACAACAA	1-10-1MOE	1073
147746	65297	65308	TAAAAACAACAA	1-10-1MOE	1073
						147689	65302	65313	CAGAGAAGGTCT	1-10-1MOE	987
147689	65448	65459	CAGAGAAGGTCT	1-10-1MOE	987
						147717	65862	65873	ATCTTCAGAGAT	1-10-1MOE	996
147717	65895	65906	ATCTTCAGAGAT	1-10-1MOE	996
						147729	66000	66011	GTAAGAGGCAGG	1-10-1MOE	920
147717	66008	66019	ATCTTCAGAGAT	1-10-1MOE	996
						147717	66041	66052	ATCTTCAGAGAT	1-10-1MOE	996
147708	66046	66057	TTGATATAGTCA	1-10-1MOE	997
						147718	66055	66066	TAATATGACTTG	1-10-1MOE	998
147729	66146	66157	GTAAGAGGCAGG	1-10-1MOE	920
						147089	66236	66247	TCCCTCTACACC	1-10-1MOE	956
368363	66281	66294	CTTAGAAGGCAGCA	2-10-2MOE	1114
						147727	66293	66304	CAGTGGACCACA	1-10-1MOE	1128
147093	66319	66330	TTGTTCCCTCTA	1-10-1MOE	929
						147094	66320	66331	GTTGTTCCCTCT	1-10-1MOE	1115
147089	66382	66393	TCCCTCTACACC	1-10-1MOE	956
						368363	66427	66440	CTTAGAAGGCAGCA	2-10-2MOE	1114
147727	66439	66450	CAGTGGACCACA	1-10-1MOE	1128
						147719	66441	66452	CCAACTCCAACT	1-10-1MOE	1116
147093	66465	66476	TTGTTCCCTCTA	1-10-1MOE	929
						147094	66466	66477	GTTGTTCCCTCT	1-10-1MOE	1115
147075	66561	66572	TCCACTGATCCT	1-10-1MOE	1026
						368357	66562	66575	CCTTCCACTGATCC	2-10-2MOE	1046
147076	66562	66573	TTCCACTGATCC	1-10-1MOE	1029
						368377	66562	66577	CTCCTTCCACTGATCC	3-10-3MOE	1030
147077	66563	66574	CTTCCACTGATC	1-10-1MOE	1047
						368358	66563	66576	TCCTTCCACTGATC	2-10-2MOE	1031
147078	66564	66575	CCTTCCACTGAT	1-10-1MOE	1044
						147079	66565	66576	TCCTTCCACTGA	1-10-1MOE	1001
147080	66566	66577	CTCCTTCCACTG	1-10-1MOE	1021
						147081	66567	66578	GCTCCTTCCACT	1-10-1MOE	1006

147719	66587	66598	CCAACTCCAACT	1-10-1MOE	1116
						147075	66707	66718	TCCACTGATCCT	1-10-1MOE	1026
368377	66708	66723	CTCCTTCCACTGATCC	3-10-3MOE	1030
						147076	66708	66719	TTCCACTGATCC	1-10-1MOE	1029
368357	66708	66721	CCTTCCACTGATCC	2-10-2MOE	1046
						147077	66709	66720	CTTCCACTGATC	1-10-1MOE	1047
147078	66710	66721	CCTTCCACTGAT	1-10-1MOE	1044
						147079	66711	66722	TCCTTCCACTGA	1-10-1MOE	1001
147080	66712	66723	CTCCTTCCACTG	1-10-1MOE	1021
						147081	66713	66724	GCTCCTTCCACT	1-10-1MOE	1006
147089	66842	66853	TCCCTCTACACC	1-10-1MOE	956
						147089	66988	66999	TCCCTCTACACC	1-10-1MOE	956
147075	66999	67010	TCCACTGATCCT	1-10-1MOE	1026
						147075	67145	67156	TCCACTGATCCT	1-10-1MOE	1026
147705	67213	67224	CGGTTTTTGTTC	1-10-1MOE	1002
						401413	67301	67314	TGCAGCCATGTACT	2-10-2MOE	1022
147737	67309	67320	ACAGCCAGGTAG	1-10-1MOE	1067
						147080	67430	67441	CTCCTTCCACTG	1-10-1MOE	1021
147737	67455	67466	ACAGCCAGGTAG	1-10-1MOE	1067
						147080	67576	67587	CTCCTTCCACTG	1-10-1MOE	1021
147082	67578	67589	AGCTCCTTCCAC	1-10-1MOE	1036
						147090	67582	67593	TTCCCTCTACAC	1-10-1MOE	955
147091	67583	67594	GTTCCCTCTACA	1-10-1MOE	1004
						147742	67591	67602	AACTTCAGTGTC	1-10-1MOE	1041
147090	67728	67739	TTCCCTCTACAC	1-10-1MOE	955
						147698	68036	68047	CCCGCCACCACC	1-10-1MOE	928
147698	68182	68193	CCCGCCACCACC	1-10-1MOE	928
						147681	68267	68278	ATGTCATTAAAC	1-10-1MOE	965
147721	68386	68397	AATGCAGGATCT	1-10-1MOE	1118
						147681	68413	68424	ATGTCATTAAAC	1-10-1MOE	965
147712	68527	68538	ACACCATCTCCC	1-10-1MOE	1005
						147721	68532	68543	AATGCAGGATCT	1-10-1MOE	1118
147711	68760	68771	AAGGGCCCTGGG	1-10-1MOE	1040
						147711	68906	68917	AAGGGCCCTGGG	1-10-1MOE	1040
147696	69045	69056	TGGATGATTGGC	1-10-1MOE	906
						147696	69191	69202	TGGATGATTGGC	1-10-1MOE	906
147723	69194	69205	GACTCCAAAGTC	1-10-1MOE	892
						147723	69210	69221	GACTCCAAA GTC	1-10-1MOE	892
389965	69271	69282	CTGCAACATGAT	1-10-1MOE	1018
						389764	69271	69282	CTGCAACATGAT	1-9-2MOE	1018
147723	69340	69351	GACTCCAAAGTC	1-10-1MOE	892
						147723	69356	69367	GACTCCAAAGTC	1-10-1MOE	892
398101	69357	69370	TTTGATAAAGCCCT	2-10-2MOE	1064
						389965	69417	69428	CTGCAACATGAT	1-10-1MOE	1018
389764	69417	69428	CTGCAACATGAT	1-9-2MOE	1018
						398101	69503	69516	TTTGATAAAGCCCT	2-10-2MOE	1064
368353	69519	69532	CACTGATCCTGCAC	2-10-2MOE	1007
						147074	69522	69533	CCACTGATCCTG	1-10-1MOE	845
147081	69631	69642	GCTCCTTCCACT	1-10-1MOE	1006
						368353	69665	69678	CACTGATCCTGCAC	2-10-2MOE	1007
147720	69729	69740	GATCTCTCGAGT	1-10-1MOE	1117
						147721	69736	69747	AATGCAGGATCT	1-10-1MOE	1118
398167	69757	69768	CAGGCCATGTGG	1-10-1MOE	1059
						147722	69762	69773	AAAGTCAGGCCA	1-10-1MOE	1130

147723	69768	69779	GACTCCAAA GTC	1-10-1MOE	892
						147080	69776	69787	CTCCTTCCACTG	1-10-1MOE	1021
147081	69777	69788	GCTCCTTCCACT	1-10-1MOE	1006
						398093	69811	69824	TCGGACTTTGAAAA	2-10-2MOE	1009
398168	69813	69824	TCGGACTTTGAA	1-10-1MOE	1008
						147725	69814	69825	CTCGGACTTTGA	1-10-1MOE	1119
147726	69819	69830	TGACTCTCGGAC	1-10-1MOE	1120
						147727	69860	69871	CAGTGGACCACA	1-10-1MOE	1128
147720	69875	69886	GATCTCTCGAGT	1-10-1MOE	1117
						147721	69882	69893	AATGCAGGATCT	1-10-1MOE	1118
147728	69899	69910	GCCAGACAGAAG	1-10-1MOE	1013
						398094	69901	69914	ATCAGCCAGACAGA	2-10-2MOE	1010
398167	69903	69914	CAGGCCATGTGG	1-10-1MOE	1059
						398092	69904	69917	AGTCAGGCCATGTG	2-10-2MOE	1060
147722	69908	69919	AAAGTCAGGCCA	1-10-1MOE	1130
						147723	69914	69925	GACTCCAAAGTC	1-10-1MOE	892
147729	69916	69927	GTAAGAGGCAGG	1-10-1MOE	920
						398095	69919	69932	CATCAGCAAGAGGC	2-10-2MOE	1011
398093	69957	69970	TCGGACTTTGAAAA	2-10-2MOE	1009
						398168	69959	69970	TCGGACTTTGAA	1-10-1MOE	1008
147725	69960	69971	CTCGGACTTTGA	1-10-1MOE	1119
						147726	69965	69976	TGACTCTCGGAC	1-10-1MOE	1120
147704	69991	70002	TTGTTCTTAGGA	1-10-1MOE	1012
						147727	70006	70017	CAGTGGACCACA	1-10-1MOE	1128
147728	70045	70056	GCCAGACAGAAG	1-10-1MOE	1013
						398094	70047	70060	ATCAGCCAGACAGA	2-10-2MOE	1010
398169	70048	70059	TCAGCCAGACAG	1-10-1MOE	909
						147729	70062	70073	GTAAGAGGCAGG	1-10-1MOE	920
398095	70065	70078	CATCAGCAAGAGGC	2-10-2MOE	1011
						147704	70137	70148	TTGTTCTTAGGA	1-10-1MOE	1012
147697	70161	70172	CCCCAGCAGCGG	1-10-1MOE	1000
						147697	70307	70318	CCCCAGCAGCGG	1-10-1MOE	1000
147728	70450	70461	GCCAGACAGAAG	1-10-1MOE	1013
						398164	70464	70475	TTGTCGATCTGC	1-10-1MOE	1014
147730	70465	70476	CTTGTCCATCAG	1-10-1MOE	1121
						147731	70471	70482	TTTCCTCTTGTC	1-10-1MOE	934
147732	70476	70487	GGGTCTTTCCTC	1-10-1MOE	1122
						147733	70497	70508	TTCTTGATGTCC	1-10-1MOE	891
398096	70562	70575	GGAGAAGCGCAGCT	2-10-2MOE	1015
						147735	70564	70575	GGAGAAGCGCAG	1-10-1MOE	1016
147736	70569	70580	AGGTAGGAGAAG	1-10-1MOE	963
						147737	70575	70586	ACAGCCAGGTAG	1-10-1MOE	1067
147728	70596	70607	GCCAGACAGAAG	1-10-1MOE	1013
						398164	70610	70621	TTGTCGATCTGC	1-10-1MOE	1014
147730	70611	70622	CTTGTCCATCAG	1-10-1MOE	1121
						368349	70616	70629	CTGCACTGACGAGT	2-10-2MOE	1017
147731	70617	70628	TTTCCTCTTGTC	1-10-1MOE	934
						147732	70622	70633	GGGTCTTTCCTC	1-10-1MOE	1122
147733	70643	70654	TTCTTGATGTCC	1-10-1MOE	891
						398096	70708	70721	GGAGAAGCGCAGCT	2-10-2MOE	1015
147735	70710	70721	GGAGAAGCGCAG	1-10-1MOE	1016
						147736	70715	70726	AGGTAGGAGAAG	1-10-1MOE	963
147737	70721	70732	ACAGCCAGGTAG	1-10-1MOE	1067
						389764	70784	70795	CTGCAACATGAT	1-9-2MOE	1018

389965	70784	70795	CTGCAACATGAT	1-10-1MOE	1018
						389965	70930	70941	CTGCAACATGAT	1-10-1MOE	1018
389764	70930	70941	CTGCAACATGAT	1-9-2MOE	1018
						368386	70995	71010	CACTGATCCTTAGAAG	3-10-3MOE	1123
368367	70997	71010	CACTGATCCTTAGA	2-10-2MOE	1124
						368387	70997	71012	TCCACTGATCCTTAGA	3-10-3MOE	1125
368354	70999	71012	TCCACTGATCCTGC	2-10-2MOE	1024
						368374	70999	71014	CTTCCACTGATCCTGC	3-10-3MOE	1126
368368	70999	71012	TCCACTGATCCTTA	2-10-2MOE	1127
						368388	70999	71014	CTTCCACTGATCCTTA	3-10-3MOE	895
368355	71000	71013	TTCCACTGATCCTG	2-10-2MOE	1025
						147074	71000	71011	CCACTGATCCTG	1-10-1MOE	845
368375	71000	71015	CCTTCCACTGATCCTG	3-10-3MOE	1020
						147075	71001	71012	TCCACTGATCCT	1-10-1MOE	1026
368376	71001	71016	TCCTTCCACTGATCCT	3-10-3MOE	1028
						147076	71002	71013	TTCCACTGATCC	1-10-1MOE	1029
368357	71002	71015	CCTTCCACTGATCC	2-10-2MOE	1046
						368377	71002	71017	CTCCTTCCACTGATCC	3-10-3MOE	1030
147077	71003	71014	CTTCCACTGATC	1-10-1MOE	1047
						368378	71003	71018	GCTCCTTCCACTGATC	3-10-3MOE	1032
147078	71004	71015	CCTTCCACTGAT	1-10-1MOE	1044
						368359	71005	71018	GCTCCTTCCACTGA	2-10-2MOE	1033
368379	71005	71020	AAGCTCCTTCCACTGA	3-10-3MOE	1034
						147079	71005	71016	TCCTTCCACTGA	1-10-1MOE	1001
147080	71006	71017	CTCCTTCCACTG	1-10-1MOE	1021
						368360	71007	71020	AAGCTCCTTCCACT	2-10-2MOE	1035
368380	71007	71022	GAAAGCTCCTTCCACT	3-10-3MOE	896
						147081	71007	71018	GCTCCTTCCACT	1-10-1MOE	1006
147082	71008	71019	AGCTCCTTCCAC	1-10-1MOE	1036
						368361	71009	71022	GAAAGCTCCTTCCA	2-10-2MOE	962
368381	71009	71024	GGGAAAGCTCCTTCCA	3-10-3MOE	1037
						147738	71067	71078	TGGGTGGCCGGG	1-10-1MOE	1069
147739	71071	71082	CGTTTGGGTGGC	1-10-1MOE	1023
						147740	71088	71099	TGTGAGGCTCCA	1-10-1MOE	1062
147741	71129	71140	CACCCACTGGTG	1-10-1MOE	1055
						368366	71141	71154	CTGATCCTTAGAAG	2-10-2MOE	1019
368386	71141	71156	CACTGATCCTTAGAAG	3-10-3MOE	1123
						368367	71143	71156	CACTGATCCTTAGA	2-10-2MOE	1124
368387	71143	71158	TCCACTGATCCTTAGA	3-10-3MOE	1125
						368374	71145	71160	CTTCCACTGATCCTGC	3-10-3MOE	1126
368354	71145	71158	TCCACTGATCCTGC	2-10-2MOE	1024
						368368	71145	71158	TCCACTGATCCTTA	2-10-2MOE	1127
368388	71145	71160	CTTCCACTGATCCTTA	3-10-3MOE	895
						368355	71146	71159	TTCCACTGATCCTG	2-10-2MOE	1025
368375	71146	71161	CCTTCCACTGATCCTG	3-10-3MOE	1020
						147075	71147	71158	TCCACTGATCCT	1-10-1MOE	1026
368356	71147	71160	CTTCCACTGATCCT	2-10-2MOE	1027
						368376	71147	71162	TCCTTCCACTGATCCT	3-10-3MOE	1028
147076	71148	71159	TTCCACTGATCC	1-10-1MOE	1029
						368357	71148	71161	CCTTCCACTGATCC	2-10-2MOE	1046
368377	71148	71163	CTCCTTCCACTGATCC	3-10-3MOE	1030
						147077	71149	71160	CTTCCACTGATC	1-10-1MOE	1047
368358	71149	71162	TCCTTCCACTGATC	2-10-2MOE	1031
						368378	71149	71164	GCTCCTTCCACTGATC	3-10-3MOE	1032

147078	71150	71161	CCTTCCACTGAT	1-10-1MOE	1044
						368359	71151	71164	GCTCCTTCCACTGA	2-10-2MOE	1033
147079	71151	71162	TCCTTCCACTGA	1-10-1MOE	1001
						368379	71151	71166	AAGCTCCTTCCACTGA	3-10-3MOE	1034
147080	71152	71163	CTCCTTCCACTG	1-10-1MOE	1021
						368380	71153	71168	GAAAGCTCCTTCCACT	3-10-3MOE	896
147081	71153	71164	GCTCCTTCCACT	1-10-1MOE	1006
						368360	71153	71166	AAGCTCCTTCCACT	2-10-2MOE	1035
147082	71154	71165	AGCTCCTTCCAC	1-10-1MOE	1036
						368381	71155	71170	GGGAAAGCTCCTTCCA	3-10-3MOE	1037
368361	71155	71168	GAAAGCTCCTTCCA	2-10-2MOE	962
						398097	71158	71171	GGCAGTCTTTATCC	2-10-2MOE	897
147738	71213	71224	TGGGTGGCCGGG	1-10-1MOE	1069
						147739	71217	71228	CGTTTGGGTGGC	1-10-1MOE	1023
147740	71234	71245	TGTGAGGCTCCA	1-10-1MOE	1062
						147741	71275	71286	CACCCACTGGTG	1-10-1MOE	1055
398097	71304	71317	GGCAGTCTTTATCC	2-10-2MOE	897
						147727	71702	71713	CAGTGGACCACA	1-10-1MOE	1128
147727	71848	71859	CAGTGGACCACA	1-10-1MOE	1128
						390030	71986	71997	TTTATAAAACTG	1-10-1MOE	1074
147102	72015	72026	TGCGAGTTGTTG	1-10-1MOE	1129
						390030	72132	72143	TTTATAAAACTG	1-10-1MOE	1074
147102	72161	72172	TGCGAGTTGTTG	1-10-1MOE	1129
						147722	72199	72210	AAAGTCAGGCCA	1-10-1MOE	1130
147696	72232	72243	TGGATGATTGGC	1-10-1MOE	906
						147741	72254	72265	CACCCACTGGTG	1-10-1MOE	1055
147722	72345	72356	AAAGTCAGGCCA	1-10-1MOE	1130
						147696	72378	72389	TGGATGATTGGC	1-10-1MOE	906
147741	72400	72411	CACCCACTGGTG	1-10-1MOE	1055
						147711	72446	72457	AAGGGCCCTGGG	1-10-1MOE	1040
398098	72574	72587	TAACTTCAGTGTCT	2-10-2MOE	1131
						147742	72575	72586	AACTTCAGTGTC	1-10-1MOE	1041
147698	72595	72606	CCCGCCACCACC	1-10-1MOE	928
						147743	72690	72701	AGGGCTTCCAGT	1-10-1MOE	1042
398099	72690	72703	GAAGGGCTTCCAGT	2-10-2MOE	1132
						147744	72694	72705	AGGAAGGGCTTC	1-10-1MOE	1043
398100	72697	72710	TGACCAGGAAGGGC	2-10-2MOE	1133
						147745	72700	72711	TTGACCAGGAAG	1-10-1MOE	1058
398098	72720	72733	TAACTTCAGTGTCT	2-10-2MOE	1131
						147742	72721	72732	AACTTCAGTGTC	1-10-1MOE	1041
147698	72741	72752	CCCGCCACCACC	1-10-1MOE	928
						398157	72757	72770	GGAAACATACCCTG	2-10-2MOE	1045
147743	72836	72847	AGGGCTTCCAGT	1-10-1MOE	1042
						398099	72836	72849	GAAGGGCTTCCAGT	2-10-2MOE	1132
147744	72840	72851	AGGAAGGGCTTC	1-10-1MOE	1043
						398100	72843	72856	TGACCAGGAAGGGC	2-10-2MOE	1133
147745	72846	72857	TTGACCAGGAAG	1-10-1MOE	1058
						147076	72898	72909	TTCCACTGATCC	1-10-1MOE	1029
368357	72898	72911	CCTTCCACTGATCC	2-10-2MOE	1046
						147077	72899	72910	CTTCCACTGATC	1-10-1MOE	1047
147078	72900	72911	CCTTCCACTGAT	1-10-1MOE	1044
						398157	72903	72916	GGAAACATACCCTG	2-10-2MOE	1045
398158	72983	72996	AGGCCCTGAGATTA	2-10-2MOE	1134
						398159	72988	73001	GGTTAAGGCCCTGA	2-10-2MOE	1135

398160	72993	73006	GAATAGGTTAAGGC	2-10-2MOE	1048
						147076	73044	73055	TTCCACTGATCC	1-10-1MOE	1029
368357	73044	73057	CCTTCCACTGATCC	2-10-2MOE	1046
						147077	73045	73056	CTTCCACTGATC	1-10-1MOE	1047
147078	73046	73057	CCTTCCACTGAT	1-10-1MOE	1044
						147746	73052	73063	TAAAAACAACAA	1-10-1MOE	1073
398161	73092	73105	AACAATGTGTTGTA	2-10-2MOE	1049
						147746	73101	73112	TAAAAACAACAA	1-10-1MOE	1073
398158	73129	73142	AGGCCCTGAGATTA	2-10-2MOE	1134
						398159	73134	73147	GGTTAAGGCCCTGA	2-10-2MOE	1135
398160	73139	73152	GAATAGGTTAAGGC	2-10-2MOE	1048
						147746	73198	73209	TAAAAACAACAA	1-10-1MOE	1073
398161	73238	73251	AACAATGTGTTGTA	2-10-2MOE	1049
						147746	73247	73258	TAAAAACAACAA	1-10-1MOE	1073
147088	73273	73284	CCCTCTACACCA	1-10-1MOE	1050
						398105	73401	73414	TGCACAGGCAGGTT	2-10-2MOE	1066
398105	73547	73560	TGCACAGGCAGGTT	2-10-2MOE	1066
						147741	73559	73570	CACCCACTGGTG	1-10-1MOE	1055
147741	73705	73716	CACCCACTGGTG	1-10-1MOE	1055
						398162	73968	73981	ACCAAACAGTTCAG	2-10-2MOE	1057
147745	73991	74002	TTGACCAGGAAG	1-10-1MOE	1058
						398167	74008	74019	CAGGCCATGTGG	1-10-1MOE	1059
398092	74009	74022	AGTCAGGCCATGTG	2-10-2MOE	1060
						398162	74114	74127	ACCAAACAGTTCAG	2-10-2MOE	1057
147745	74137	74148	TTGACCAGGAAG	1-10-1MOE	1058
						398167	74154	74165	CAGGCCATGTGG	1-10-1MOE	1059
147089	74280	74291	TCCCTCTACACC	1-10-1MOE	956
						147090	74281	74292	TTCCCTCTACAC	1-10-1MOE	955
389949	74310	74321	GCGCGAGCCCGA	1-10-1MOE	1061
						147740	74339	74350	TGTGAGGCTCCA	1-10-1MOE	1062
389950	74381	74392	CCCTGAAGGTTC	1-10-1MOE	1063
						147089	74426	74437	TCCCTCTACACC	1-10-1MOE	956
147090	74427	74438	TTCCCTCTACAC	1-10-1MOE	955
						389949	74456	74467	GCGCGAGCCCGA	1-10-1MOE	1061
147685	74490	74501	GGCTGACATTCA	1-10-1MOE	975
						398101	74510	74523	TTTGATAAAGCCCT	2-10-2MOE	1064
398102	74536	74549	CTACCTGAGGATTT	2-10-2MOE	899
						398103	74543	74556	CCCAGTACTACCTG	2-10-2MOE	900
147685	74636	74647	GGCTGACATTCA	1-10-1MOE	975
						398102	74682	74695	CTACCTGAGGATTT	2-10-2MOE	899
398103	74689	74702	CCCAGTACTACCTG	2-10-2MOE	900
						147736	74737	74748	AGGTAGGAGAAG	1-10-1MOE	963
398104	74805	74818	CAAGAAGACCTTAC	2-10-2MOE	1065
						147736	74883	74894	AGGTAGGAGAAG	1-10-1MOE	963
147737	74893	74904	ACAGCCAGGTAG	1-10-1MOE	1067
						398105	74894	74907	TGCACAGGCAGGTT	2-10-2MOE	1066
147737	74919	74930	ACAGCCAGGTAG	1-10-1MOE	1067
						398095	74940	74953	CATCAGCAAGAGGC	2-10-2MOE	1011
398104	74951	74964	CAAGAAGACCTTAC	2-10-2MOE	1065
						398106	74974	74987	TGGAAAACTGCACC	2-10-2MOE	1068
398107	74980	74993	TATTCCTGGAAAAC	2-10-2MOE	902
						147745	75030	75041	TTGACCAGGAAG	1-10-1MOE	1058
147737	75039	75050	ACAGCCAGGTAG	1-10-1MOE	1067
						398105	75040	75053	TGCACAGGCAGGTT	2-10-2MOE	1066

147737	75065	75076	ACAGCCAGGTAG	1-10-1MOE	1067
						398108	75077	75090	GGAATGTCTGAGTT	2-10-2MOE	1136
398095	75086	75099	CATCAGCAAGAGGC	2-10-2MOE	1011
						147691	75108	75119	GAGGTGGGAAAA	1-10-1MOE	966
398106	75120	75133	TGGAAAACTGCACC	2-10-2MOE	1068
						398107	75126	75139	TATTCCTGGAAAAC	2-10-2MOE	902
147738	75155	75166	TGGGTGGCCGGG	1-10-1MOE	1069
						147745	75176	75187	TTGACCAGGAAG	1-10-1MOE	1058
398108	75223	75236	GGAATGTCTGAGTT	2-10-2MOE	1136
						398109	75247	75260	CAAGAAGTGTGGTT	2-10-2MOE	903
147691	75254	75265	GAGGTGGGAAAA	1-10-1MOE	966
						147738	75301	75312	TGGGTGGCCGGG	1-10-1MOE	1069
398110	75385	75398	GTTCCCTTTGCAGG	2-10-2MOE	952
						147091	75387	75398	GTTCCCTCTACA	1-10-1MOE	1004
398109	75393	75406	CAAGAAGTGTGGTT	2-10-2MOE	903
						398111	75470	75483	GTGAAAATGCTGGC	2-10-2MOE	904
401385	75494	75507	CCCAGTGGGTTTGA	2-10-2MOE	890
						398166	75499	75510	GGGCTTCTTCCA	1-10-1MOE	1070
147091	75525	75536	GTTCCCTCTACA	1-10-1MOE	1004
						147092	75526	75537	TGTTCCCTCTAC	1-10-1MOE	901
398110	75531	75544	GTTCCCTTTGCAGG	2-10-2MOE	952
						147091	75533	75544	GTTCCCTCTACA	1-10-1MOE	1004
147706	75540	75551	GCTGA CATCTCG	1-10-1MOE	1071
						398112	75584	75597	CAGCCTGGCACCTA	2-10-2MOE	1072
398111	75616	75629	GTGAAAATGCTGGC	2-10-2MOE	904
						147746	75617	75628	TAAAAACAACAA	1-10-1MOE	1073
398166	75645	75656	GGGCTTCTTCCA	1-10-1MOE	1070
						147091	75671	75682	GTTCCCTCTACA	1-10-1MOE	1004
147092	75672	75683	TGTTCCCTCTAC	1-10-1MOE	901
						398113	75693	75706	AGGAGGTTAAACCA	2-10-2MOE	905
398112	75730	75743	CAGCCTGGCACCTA	2-10-2MOE	1072
						147746	75763	75774	TAAAAACAACAA	1-10-1MOE	1073
398114	75770	75783	AGGCATATAGCAGA	2-10-2MOE	1075
						398115	75786	75799	AGTAAATATTGGCT	2-10-2MOE	1076
398116	75799	75812	TAATGACCTGATGA	2-10-2MOE	1137
						398113	75839	75852	AGGAGGTTAAACCA	2-10-2MOE	905
390030	75839	75850	TTTATAAAACTG	1-10-1MOE	1074
						398115	75932	75945	AGTAAATATTGGCT	2-10-2MOE	1076
398116	75945	75958	TAATGACCTGATGA	2-10-2MOE	1137
						398106	75982	75995	TGGAAAACTGCACC	2-10-2MOE	1068
390030	75985	75996	TTTATAAAACTG	1-10-1MOE	1074
						398106	76127	76140	TGGAAAACTGCACC	2-10-2MOE	1068
147690	76196	76207	TGAAGTTAATTC	1-10-1MOE	1138
						147690	76341	76352	TGAAGTTAATTC	1-10-1MOE	1138
147724	76740	76751	GAAATTGAGGAA	1-10-1MOE	1139
						147089	76873	76884	TCCCTCTACACC	1-10-1MOE	956
147679	76881	76892	CAAAAGGATCCC	1-10-1MOE	907
						147724	76885	76896	GAAATTGAGGAA	1-10-1MOE	1139
147089	77018	77029	TCCCTCTACACC	1-10-1MOE	956
						147679	77026	77037	CAAAAGGATCCC	1-10-1MOE	907
147693	77240	77251	GTGCGCTCCCAT	1-10-1MOE	1078
						147697	77759	77770	CCCCAGCAGCGG	1-10-1MOE	1000

In certain embodiments, the target zone is the nucleotide 177-190 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 177-190 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 177-190 comprises the nucleotide sequence that is selected from SEQ ID NO 886,859 or 853.In some this embodiment, the short antisense compounds of the nucleotide 177-190 of targeting SEQ ID NO:11 is selected from Isis No 147022,147023 or 147024.

In certain embodiments, the target zone is the nucleotide 195-228 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 195-228 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 195-228 comprises the nucleotide sequence that is selected from SEQ ID NO 877,868,882,886,859,853,865,835,843,846,842,848,874,849,863,855,850,864 or 834.In some this embodiment, the short antisense compounds of the nucleotide 195-228 of targeting SEQ ID NO:11 is selected from Isis No 147019,147020,147021,147022,147023,147024,147025,147026,147027,147028,147073,147029,147030,147036,147037,147038,147039,147040 or 147041.

In certain embodiments, the target zone is the nucleotide 323-353 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 323-353 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 323-353 comprises the nucleotide sequence that is selected from SEQ ID NO 866,881,869,883,858,833,875,837,829,871,884,887,839,830,840,861 or 879.In some this embodiment, the short antisense compounds of the nucleotide 323-353 of targeting SEQ ID NO:11 is selected from Isis No 147042,147043,147044,147045,147046,147047,147051,147052,147053,147054,147055,147056,147057,147058,147059,147060 or 147061.

In certain embodiments, the target zone is the nucleotide 322-353 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 177-190 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 322-353 comprises the nucleotide sequence that is selected from SEQ ID NO 842,866,881,869,883,858,833,875,837,829,871,884,887,839,830,840,861 or 879.In some this embodiment, the short antisense compounds of the nucleotide 322-353 of targeting SEQ ID NO:11 is selected from Isis No 147073,147042,147043,147044,147045,147046,147047,147051,147052,147053,147054,147055,147056,147057,147058,147059,147060 or 147061.

In certain embodiments, the target zone is the nucleotide 679-799 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 679-799 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 679-799 comprises the nucleotide sequence that is selected from SEQ ID NO 883,858,883 or 858.In some this embodiment, the short antisense compounds of the nucleotide 679-799 of targeting SEQ IDNO:11 is selected from Isis No 147045,147046,147045 or 147046.

In certain embodiments, the target zone is the nucleotide 679-827 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 679-827 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 679-827 comprises the nucleotide sequence that is selected from SEQ ID NO 883,858,883,858 or 851.In some this embodiment, the short antisense compounds of the nucleotide 679-827 of targeting SEQ ID NO:11 is selected from Isis No 147045,147046,147045,147046 or 147066.

In certain embodiments, the target zone is the nucleotide 1024-1046 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 1024-1046 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 1024-1046 comprises the nucleotide sequence that is selected from SEQ ID NO 841,862,880,857,851,876,838,860,878,856,832 or 842.In some this embodiment, the short antisense compounds of the nucleotide 1024-1046 of targeting SEQ ID NO:11 is selected from Isis No 147062,147063,147064,147065,147066,147067,147068,147069,147070,147071,147072 or 147073.

In certain embodiments, the target zone is the nucleotide 992-1046 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 992-1046 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 992-1046 comprises the nucleotide sequence that is selected from SEQ ID NO 831,841,862,880,857,851,876,838,860,878,856,832 or 842.In some this embodiment, the short antisense compounds of the nucleotide 992-1046 of targeting SEQ ID NO:11 is selected from Isis No 404131,147062,147063,147064,147065,147066,147067,147068,147069,147070,147071,147072 or 147073.

In certain embodiments, the target zone is the nucleotide 1868-1881 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 1868-1881 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 1868-1881 comprises the nucleotide sequence that is selected from SEQ ID NO 886,859 or 853.In some this embodiment, the short antisense compounds of the nucleotide 1868-1881 of targeting SEQ ID NO:11 is selected from Isis No 147022,147023 or 147024.

In certain embodiments, the target zone is the nucleotide 1886-1919 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 1886-1919 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 1886-1919 comprises the nucleotide sequence that is selected from SEQ ID NO 877,868,882,886,859,865,843,846,874,863,855,864 or 834.In some this embodiment, the short antisense compounds of the nucleotide 1886-1919 of targeting SEQ ID NO:11 is selected from Isis No 147019,147020,147021,147022,147023,147025,147027,147028,147030,147037,147038,147040 or 147041.

In certain embodiments, the target zone is the nucleotide 1869-1919 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 1869-1919 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 1869-1919 comprises the nucleotide sequence that is selected from SEQ ID NO 859,853,877,868,882,886,859,865,843,846,874,863,855,864 or 834.In some this embodiment, the short antisense compounds of the nucleotide 1869-1919 of targeting SEQ ID NO:11 is selected from Isis No 147023,147024,147019,147020,147021,147022,147023,147025,147027,147028,147030,147037,147038,147040 or 147041.

In certain embodiments, the target zone is the nucleotide 1976-1989 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 1976-1989 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 1976-1989 comprises the nucleotide sequence that is selected from SEQ ID NO 886,859 or 853.In some this embodiment, the short antisense compounds of the nucleotide 1976-1989 of targeting SEQ ID NO:11 is selected from Isis No 147022,147023 or 147024.

In certain embodiments, the target zone is the nucleotide 1995-2027 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 1995-2027 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 1995-2027 comprises the nucleotide sequence that is selected from SEQ ID NO 868,882,886,859,853,865,835,843,846,848,874,849,863,855,850,864 or 834.In some this embodiment, the short antisense compounds of the nucleotide 1995-2027 of targeting SEQ ID NO:11 is selected from Isis No 147020,147021,147022,147023,147024,147025,147026,147027,147028,147029,147030,147036,147037,147038,147039,147040 or 147041.

In certain embodiments, the target zone is the nucleotide 2366-2382 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 2366-2382 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 2366-2382 comprises the nucleotide sequence that is selected from SEQ ID NO 867 or 873.In some this embodiment, the short antisense compounds of the nucleotide 2366-2382 of targeting SEQ ID NO:11 is selected from Isis No 404199 or 404134.

In certain embodiments, the target zone is the nucleotide 6220-6233 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 6220-6233 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 6220-6233 comprises the nucleotide sequence that is selected from SEQ ID NO 870,836 or 844.In some this embodiment, the short antisense compounds of the nucleotide 6220-6233 of targeting SEQ ID NO:11 is selected from Isis No 147032,147033 or 147034.

In certain embodiments, the target zone is the nucleotide 6288-6300 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 6288-6300 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 6288-6300 comprises the nucleotide sequence that is selected from SEQ ID NO 869 or 883.In some this embodiment, the short antisense compounds of the nucleotide 6288-6300 of targeting SEQ ID NO:11 is selected from Isis No 147044 or 147045.

In certain embodiments, the target zone is the nucleotide 6329-6342 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 6329-6342 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 6329-6342 comprises the nucleotide sequence that is selected from SEQ ID NO 870,836 or 844.In some this embodiment, the short antisense compounds of the nucleotide 6329-6342 of targeting SEQ ID NO:11 is selected from Isis No 147032,147033 or 147034.

In certain embodiments, the target zone is the nucleotide 6397-6409 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 6397-6409 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 6397-6409 comprises the nucleotide sequence that is selected from SEQ ID NO 869 or 883.In some this embodiment, the short antisense compounds of the nucleotide 6397-6409 of targeting SEQ ID NO:11 is selected from Isis No 147044 or 147045.

In certain embodiments, the target zone is the nucleotide 7057-7178 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 7057-7178 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 7057-7178 comprises the nucleotide sequence that is selected from SEQ ID NO 830,840,861,830 or 840.In some this embodiment, the short antisense compounds of the nucleotide 7057-7178 of targeting SEQ ID NO:11 is selected from Isis No 147058,147059,147060,147058 or 147059.

In certain embodiments, the target zone is the nucleotide 8630-8750 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 8630-8750 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 8630-8750 comprises the nucleotide sequence that is selected from SEQ ID NO 843,846,843 or 846.In some this embodiment, the short antisense compounds of the nucleotide 8630-8750 of targeting SEQ ID NO:11 is selected from Isis No 147027,147028,147027 or 147028.

In certain embodiments, the target zone is the nucleotide 10957-11077 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 10957-11077 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 10957-11077 comprises the nucleotide sequence that is selected from SEQ ID NO 881,869,881 or 869.In some this embodiment, the short antisense compounds of the nucleotide 10957-11077 of targeting SEQ ID NO:11 is selected from Isis No 147043,147044,147043 or 147044.

In certain embodiments, the target zone is the nucleotide 11605-11623 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 11605-11623 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 11605-11623 comprises the nucleotide sequence that is selected from SEQ ID NO 856,878 or 856.In some this embodiment, the short antisense compounds of the nucleotide 11605-11623 of targeting SEQ ID NO:11 is selected from Isis No 147071,147070 or 147071.

In certain embodiments, the target zone is the nucleotide 12805-12817 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 12805-12817 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 12805-12817 comprises the nucleotide sequence that is selected from SEQ ID NO 874 or 885.In some this embodiment, the short antisense compounds of the nucleotide 12805-12817 of targeting SEQ ID NO:11 is selected from Isis No 147030 or 147031.

In certain embodiments, the target zone is the nucleotide 12986-12998 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 12986-12998 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 12986-12998 comprises the nucleotide sequence that is selected from SEQ ID NO 874 or 885.In some this embodiment, the short antisense compounds of the nucleotide 12986-12998 of targeting SEQ ID NO:11 is selected from Isis No 147030 or 147031.

In certain embodiments, the target zone is the nucleotide 15560-15572 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 15560-15572 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 15560-15572 comprises the nucleotide sequence that is selected from SEQ ID NO 876 or 838.In some this embodiment, the short antisense compounds of the nucleotide 15560-15572 of targeting SEQ ID NO:11 is selected from Isis No 147067 or 147068.

In certain embodiments, the target zone is the nucleotide 17787-17941 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 17787-17941 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 17787-17941 comprises the nucleotide sequence that is selected from SEQ ID NO 874 or 880.In some this embodiment, the short antisense compounds of the nucleotide 17787-17941 of targeting SEQ ID NO:11 is selected from Isis No 147030 or 147064.

In certain embodiments, the target zone is the nucleotide 21190-21202 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 21190-21202 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 21190-21202 comprises the nucleotide sequence that is selected from SEQ ID NO 843 or 846.In some this embodiment, the short antisense compounds of the nucleotide 21190-21202 of targeting SEQ ID NO:11 is selected from Isis No 147027 or 147028.

In certain embodiments, the target zone is the nucleotide 21358-21370 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 21358-21370 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 21358-21370 comprises the nucleotide sequence that is selected from SEQ ID NO 843 or 846.In some this embodiment, the short antisense compounds of the nucleotide 21358-21370 of targeting SEQ ID NO:11 is selected from Isis No 017027 or 147028.

In certain embodiments, the target zone is the nucleotide 24318-24332 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 24318-24332 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 24318-24332 comprises the nucleotide sequence that is selected from SEQ ID NO 881,869,883 or 858.In some this embodiment, the short antisense compounds of the nucleotide 24318-24332 of targeting SEQ ID NO:11 is selected from Isis No 147043,147044,147045 or 147046.

In certain embodiments, the target zone is the nucleotide 24486-24501 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 24486-24501 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 24486-24501 comprises the nucleotide sequence that is selected from SEQ ID NO 881,869,858 or 833.In some this embodiment, the short antisense compounds of the nucleotide 24486-24501 of targeting SEQ ID NO:11 is selected from Isis No 147043,147044,147046 or 147047.

In certain embodiments, the target zone is the nucleotide 25065-25077 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 25065-25077 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 25065-25077 comprises the nucleotide sequence that is selected from SEQ ID NO 864 or 834.In some this embodiment, the short antisense compounds of the nucleotide 25065-25077 of targeting SEQ ID NO:11 is selected from Isis No 147040 or 147041.

In certain embodiments, the target zone is the nucleotide 25232-25245 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 25232-25245 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 25232-25245 comprises the nucleotide sequence that is selected from SEQ ID NO 850,864 or 834.In some this embodiment, the short antisense compounds of the nucleotide 25232-25245 of targeting SEQ ID NO:11 is selected from Isis No 147039,147040 or 147041.

In certain embodiments, the target zone is the nucleotide 25508-25523 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 25508-25523 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 25508-25523 comprises the nucleotide sequence that is selected from SEQ ID NO 839 or 879.In some this embodiment, the short antisense compounds of the nucleotide 25508-25523 of targeting SEQ ID NO:11 is selected from Isis No 147057 or 147061.

In certain embodiments, the target zone is the nucleotide 25676-28890 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 25676-28890 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 25676-28890 comprises the nucleotide sequence that is selected from SEQ ID NO 839,860 or 878.In some this embodiment, the short antisense compounds of the nucleotide 25676-28890 of targeting SEQ ID NO:11 is selected from Isis No 147057,147069 or 147070.

In certain embodiments, the target zone is the nucleotide 33056-33069 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 33056-33069 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 33056-33069 comprises the nucleotide sequence that is selected from SEQ ID NO 860,878 or 856.In some this embodiment, the short antisense compounds of the nucleotide 33056-33069 of targeting SEQ ID NO:11 is selected from Isis No 147069,147070 or 147071.

In certain embodiments, the target zone is the nucleotide 33205-33217 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 33205-33217 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 33205-33217 comprises the nucleotide sequence that is selected from SEQ ID NO 878 or 856.In some this embodiment, the short antisense compounds of the nucleotide 33205-33217 of targeting SEQ ID NO:11 is selected from Isis No 14707 or 147071.

In certain embodiments, the target zone is the nucleotide 33318-33334 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 33318-33334 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 33318-33334 comprises the nucleotide sequence that is selected from SEQ ID NO 858,854 or 875.In some this embodiment, the short antisense compounds of the nucleotide 33318-33334 of targeting SEQ ID NO:11 is selected from Isis No 147046,147049 or 147051.

In certain embodiments, the target zone is the nucleotide 33466-33482 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 33466-33482 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 33466-33482 comprises the nucleotide sequence that is selected from SEQ ID NO 858,833 or 875.In some this embodiment, the short antisense compounds of the nucleotide 33466-33482 of targeting SEQ ID NO:11 is selected from Isis No 147046,147047 or 147051.

In certain embodiments, the target zone is the nucleotide 33640-33656 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 33640-33656 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 33640-33656 comprises the nucleotide sequence that is selected from SEQ ID NO 858 or 875.In some this embodiment, the short antisense compounds of the nucleotide 33640-33656 of targeting SEQ ID NO:11 is selected from Isis No 147046 or 147051.

In certain embodiments, the target zone is the nucleotide 33788-33804 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 33788-33804 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 33788-33804 comprises the nucleotide sequence that is selected from SEQ ID NO 858 or 875.In some this embodiment, the short antisense compounds of the nucleotide 33788-33804 of targeting SEQ ID NO:11 is selected from Isis No 147046 or 147051.

In certain embodiments, the target zone is the nucleotide 35437-35449 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 35437-35449 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 35437-35449 comprises the nucleotide sequence that is selected from SEQ ID NO 840 or 861.In some this embodiment, the short antisense compounds of the nucleotide 35437-35449 of targeting SEQ ID NO:11 is selected from Isis No 147059 or 147060.

In certain embodiments, the target zone is the nucleotide 40353-40373 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 40353-40373 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 40353-40373 comprises the nucleotide sequence that is selected from SEQ ID NO 879 or 881.In some this embodiment, the short antisense compounds of the nucleotide 40353-40373 of targeting SEQ ID NO:11 is selected from Isis No 147061 or 147043.

In certain embodiments, the target zone is the nucleotide 42527-42541 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 42527-42541 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 42527-42541 comprises the nucleotide sequence that is selected from SEQ ID NO 885,870 or 844.In some this embodiment, the short antisense compounds of the nucleotide 42527-42541 of targeting SEQ ID NO:11 is selected from Isis No 147031,147032 or 147034.

In certain embodiments, the target zone is the nucleotide 42675-42689 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 42675-42689 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 42675-42689 comprises the nucleotide sequence that is selected from SEQ ID NO 885,870,836 or 844.In some this embodiment, the short antisense compounds of the nucleotide 42675-42689 of targeting SEQ ID NO:11 is selected from Isis No 147031,147032,147033 or 147034.

In certain embodiments, the target zone is the nucleotide 46313-46328 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 46313-46328 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 46313-46328 comprises the nucleotide sequence that is selected from SEQ ID NO 839,830,840 or 879.In some this embodiment, the short antisense compounds of the nucleotide 46313-46328 of targeting SEQ ID NO:11 is selected from Isis No 147057,147058,147059 or 147061.

In certain embodiments, the target zone is the nucleotide 46461-46476 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 46461-46476 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 46461-46476 comprises the nucleotide sequence that is selected from SEQ ID NO 839,840 or 879.In some this embodiment, the short antisense compounds of the nucleotide 46461-46476 of targeting SEQ ID NO:11 is selected from Isis No 147057,147059 or 147061.

In certain embodiments, the target zone is the nucleotide 48369-48381 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 48369-48381 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 48369-48381 comprises the nucleotide sequence that is selected from SEQ ID NO 842 or 845.In some this embodiment, the short antisense compounds of the nucleotide 48369-48381 of targeting SEQ ID NO:11 is selected from Isis No 147073 or 147074.

In certain embodiments, the target zone is the nucleotide 48714-48726 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 48714-48726 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 48714-48726 comprises the nucleotide sequence that is selected from SEQ ID NO 843 or 846.In some this embodiment, the short antisense compounds of the nucleotide 48714-48726 of targeting SEQ ID NO:11 is selected from Isis No 147027 or 147028.

In certain embodiments, the target zone is the nucleotide 49050-49062 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 49050-49062 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 49050-49062 comprises the nucleotide sequence that is selected from SEQ ID NO 876 or 838.In some this embodiment, the short antisense compounds of the nucleotide 49050-49062 of targeting SEQ ID NO:11 is selected from Isis No 147067 or 147068.

In certain embodiments, the target zone is the nucleotide 49672-49684 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 49672-49684 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 49672-49684 comprises the nucleotide sequence that is selected from SEQ ID NO 842 or 845.In some this embodiment, the short antisense compounds of the nucleotide 49672-49684 of targeting SEQ ID NO:11 is selected from Isis No 147073 or 147074.

In certain embodiments, the target zone is the nucleotide 52292-52304 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 52292-52304 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 52292-52304 comprises the nucleotide sequence that is selected from SEQ ID NO 849 or 863.In some this embodiment, the short antisense compounds of the nucleotide 52292-52304 of targeting SEQ ID NO:11 is selected from Isis No 147036 or 147037.

In certain embodiments, the target zone is the nucleotide 52438-52450 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 52438-52450 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 52438-52450 comprises the nucleotide sequence that is selected from SEQ ID NO 849 or 863.In some this embodiment, the short antisense compounds of the nucleotide 52438-52450 of targeting SEQ ID NO:11 is selected from Isis No 147036 or 147037.

In certain embodiments, the target zone is the nucleotide 53445-53458 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 53445-53458 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 53445-53458 comprises the nucleotide sequence that is selected from SEQ ID NO 866,881 or 869.In some this embodiment, the short antisense compounds of the nucleotide 53445-53458 of targeting SEQ ID NO:11 is selected from Isis No 147042,147043 or 147044.

In certain embodiments, the target zone is the nucleotide 53591-53604 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 53591-53604 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 53591-53604 comprises the nucleotide sequence that is selected from SEQ ID NO 866,874,881,885 or 869.In some this embodiment, the short antisense compounds of the nucleotide 53591-53604 of targeting SEQ ID NO:11 is selected from Isis No 147042,147030,147043,147031 or 147044.

In certain embodiments, the target zone is the nucleotide 53738-53750 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 53738-53750 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 53738-53750 comprises the nucleotide sequence that is selected from SEQ ID NO 874 or 885.In some this embodiment, the short antisense compounds of the nucleotide 53738-53750 of targeting SEQ ID NO:11 is selected from Isis No 147030 or 147031.

In certain embodiments, the target zone is the nucleotide 53783-53795 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 53783-53795 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 53783-53795 comprises the nucleotide sequence that is selected from SEQ ID NO 864 or 834.In some this embodiment, the short antisense compounds of the nucleotide 53783-53795 of targeting SEQ ID NO:11 is selected from Isis No 147040 or 147041.

In certain embodiments, the target zone is the nucleotide 55008-55020 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 55008-55020 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 55008-55020 comprises the nucleotide sequence that is selected from SEQ ID NO 866 or 881.In some this embodiment, the short antisense compounds of the nucleotide 55008-55020 of targeting SEQ ID NO:11 is selected from Isis No 147042 or 147043.

In certain embodiments, the target zone is the nucleotide 55154-55166 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 55154-55166 of antisense compounds targeting SEQ ID NO:11.In some this embodiment, the short antisense compounds of targeted nucleotide 55154-55166 comprises the nucleotide sequence that is selected from SEQ ID NO 866 or 881.In some this embodiment, the short antisense compounds of the nucleotide 55154-55166 of targeting SEQ ID NO:11 is selected from Isis No 147042 or 147043.

In certain embodiments, the target zone is the nucleotide 55682-55695 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 55682-55695 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 55682-55695 comprises the nucleotide sequence that is selected from SEQ ID NO 877 or 882.In some this embodiment, the short antisense compounds of the nucleotide 55682-55695 of targeting SEQ ID NO:11 is selected from Isis No 147019 or 147021.

In certain embodiments, the target zone is the nucleotide 56275-56293 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 56275-56293 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 56275-56293 comprises the nucleotide sequence that is selected from SEQ ID NO 871,884,887,830,840,861 or 879.In some this embodiment, the short antisense compounds of the nucleotide 56275-56293 of targeting SEQ ID NO:11 is selected from Isis No 147054,147055,147056,147058,147059,147060 or 147061.

In certain embodiments, the target zone is the nucleotide 56418-56439 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 56418-56439 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 56418-56439 comprises the nucleotide sequence that is selected from SEQ ID NO 875,829,871,884,887,839,830 or 879.In some this embodiment, the short antisense compounds of the nucleotide 56418-56439 of targeting SEQ ID NO:11 is selected from Isis No 147051,147053,147054,147055,147056,147057,147058 or 147061.

In certain embodiments, the target zone is the nucleotide 57264-57276 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 57264-57276 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 57264-57276 comprises the nucleotide sequence that is selected from SEQ ID NO 883 or 858.In some this embodiment, the short antisense compounds of the nucleotide 57264-57276 of targeting SEQ ID NO:11 is selected from Isis No 147045 or 147046.

In certain embodiments, the target zone is the nucleotide 61276-61293 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 61276-61293 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 61276-61293 comprises the nucleotide sequence that is selected from SEQ ID NO 856,847,849,863,855,850 or 864.In some this embodiment, the short antisense compounds of the nucleotide 61276-61293 of targeting SEQ ID NO:11 is selected from Isis No 147071,147035,147036,147037,147038,147039 or 147040.

In certain embodiments, the target zone is the nucleotide 61257-61320 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 61257-61320 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 61257-61320 comprises the nucleotide sequence that is selected from SEQ ID NO 881,856,847,849,863,855,850,864 or 886.In some this embodiment, the short antisense compounds of the nucleotide 61257-61320 of targeting SEQ IDNO:11 is selected from Isis No 147043,147071,147035,147036,147037,147038,147039,147040 or 147071.

In certain embodiments, the target zone is the nucleotide 61422-61439 of SEQ ID NO:11.11. lack in certain embodiments, the nucleotide 61422-61439 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 61422-61439 comprises the nucleotide sequence that is selected from SEQ ID NO 844,847,849,863,855 or 864.In some this embodiment, the short antisense compounds of the nucleotide 61422-61439 of targeting SEQ IDNO:11 is selected from Isis No 147034,147035,147036,147037,147038 or 147040.

In certain embodiments, the target zone is the nucleotide 61422-61466 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 61422-61466 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 61422-61466 comprises the nucleotide sequence that is selected from SEQ ID NO 844,847,849,863,855,864 or 856.In some this embodiment, the short antisense compounds of the nucleotide 61422-61466 of targeting SEQ ID NO:11 is selected from Isis No 147034,147035,147036,147037,147038,147040 or 147071.

In certain embodiments, the target zone is the nucleotide 63065-63078 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 63065-63078 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 63065-63078 comprises the nucleotide sequence that is selected from SEQ ID NO 851 or 838.In some this embodiment, the short antisense compounds of the nucleotide 63065-63078 of targeting SEQ ID NO:11 is selected from Isis No 147066 or 147068.

In certain embodiments, the target zone is the nucleotide 63207-63222 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 63207-63222 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 63207-63222 comprises the nucleotide sequence that is selected from SEQ ID NO 841 or 851.In some this embodiment, the short antisense compounds of the nucleotide 63207-63222 of targeting SEQ ID NO:11 is selected from Isis No 147062 or 147066.

In certain embodiments, the target zone is the nucleotide 64538-64550 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 64538-64550 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 64538-64550 comprises the nucleotide sequence that is selected from SEQ ID NO 849 or 863.In some this embodiment, the short antisense compounds of the nucleotide 64538-64550 of targeting SEQ ID NO:11 is selected from Isis No 147036 or 147037.

In certain embodiments, the target zone is the nucleotide 64864-64876 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 64864-64876 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 64864-64876 comprises the nucleotide sequence that is selected from SEQ ID NO 851 or 876.In some this embodiment, the short antisense compounds of the nucleotide 64864-64876 of targeting SEQ ID NO:11 is selected from Isis No 147066 or 147067.

In certain embodiments, the target zone is the nucleotide 65010-65028 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 65010-65028 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 65010-65028 comprises the nucleotide sequence that is selected from SEQ ID NO 851,876 or 883.In some this embodiment, the short antisense compounds of the nucleotide 65010-65028 of targeting SEQ ID NO:11 is selected from Isis No 147066,147067 or 147045.

In certain embodiments, the target zone is the nucleotide 65163-65175 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 65163-65175 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 65163-65175 comprises the nucleotide sequence that is selected from SEQ ID NO 883 or 858.In some this embodiment, the short antisense compounds of the nucleotide 65163-65175 of targeting SEQ ID NO:11 is selected from Isis No 147045 or 147046.

In certain embodiments, the target zone is the nucleotide 65408-65422 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 65408-65422 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 65408-65422 comprises the nucleotide sequence that is selected from SEQ ID NO 883 or 856.In some this embodiment, the short antisense compounds of the nucleotide 65408-65422 of targeting SEQ ID NO:11 is selected from Isis No 147068 or 147071.

In certain embodiments, the target zone is the nucleotide 65549-65568 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 65549-65568 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 65549-65568 comprises the nucleotide sequence that is selected from SEQ ID NO 860,838 or 856.In some this embodiment, the short antisense compounds of the nucleotide 65549-65568 of targeting SEQ ID NO:11 is selected from Isis No 147069,147068 or 147071.

In certain embodiments, the target zone is the nucleotide 67741-67754 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 67741-67754 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 67741-67754 comprises the nucleotide sequence that is selected from SEQ ID NO 848,874 or 885.In some this embodiment, the short antisense compounds of the nucleotide 67741-67754 of targeting SEQ ID NO:11 is selected from Isis No 147029,147030 or 147031.

In certain embodiments, the target zone is the nucleotide 67886-67900 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 67886-67900 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 67886-67900 comprises the nucleotide sequence that is selected from SEQ ID NO 846,848,874 or 885.In some this embodiment, the short antisense compounds of the nucleotide 67886-67900 of targeting SEQ ID NO:11 is selected from Isis No 147028,147029,147030 or 147031.

In certain embodiments, the target zone is the nucleotide 68867-68880 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 68867-68880 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 68867-68880 comprises the nucleotide sequence that is selected from SEQ ID NO 881,869 or 883.In some this embodiment, the short antisense compounds of the nucleotide 68867-68880 of targeting SEQ ID NO:11 is selected from Isis No 147043,147044 or 147045.

In certain embodiments, the target zone is the nucleotide 69013-69532 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 69013-69532 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 69013-69532 comprises the nucleotide sequence that is selected from SEQ ID NO 881,869,883,858,856,832 or 842.In some this embodiment, the short antisense compounds of the nucleotide 69013-69532 of targeting SEQ ID NO:11 is selected from Isis No 147043,147044,147045,147046,147071,147072 or 147073.

In certain embodiments, the target zone is the nucleotide 69665-69880 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 69665-69880 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 69665-69880 comprises the nucleotide sequence that is selected from SEQ ID NO 856,832,842,845 or 851.In some this embodiment, the short antisense compounds of the nucleotide 69665-69880 of targeting SEQ ID NO:11 is selected from Isis No 147071,147072,147073,147074 or 147066.

In certain embodiments, the target zone is the nucleotide 70611-70630 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 70611-70630 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 70611-70630 comprises the nucleotide sequence that is selected from SEQ ID NO 859,841,862,880,857 or 851.In some this embodiment, the short antisense compounds of the nucleotide 70611-70630 of targeting SEQ ID NO:11 is selected from Isis No 147023,147062,147063,147064,147065 or 147066.

In certain embodiments, the target zone is the nucleotide 70762-70776 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 70762-70776 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 70762-70776 comprises the nucleotide sequence that is selected from SEQ ID NO 862,880,857 or 851.In some this embodiment, the short antisense compounds of the nucleotide 70762-70776 of targeting SEQ ID NO:11 is selected from Isis No 147063,147064,147065 or 147066.

In certain embodiments, the target zone is the nucleotide 70998-71010 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 70998-71010 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 70998-71010 comprises the nucleotide sequence that is selected from SEQ ID NO 832 or 842.In some this embodiment, the short antisense compounds of the nucleotide 70998-71010 of targeting SEQ ID NO:11 is selected from Isis No 147072 or 147073.

In certain embodiments, the target zone is the nucleotide 71144-714364 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 71144-714364 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 71144-714364 comprises the nucleotide sequence that is selected from SEQ ID NO 832,842,845,863,855 or 850.In some this embodiment, the short antisense compounds of the nucleotide 71144-714364 of targeting SEQ IDNO:11 is selected from Isis No 147072,147073,147074,147037,147038 or 147039.

In certain embodiments, the target zone is the nucleotide 71497-71652 of SEQ ID NO:11.In certain embodiments, lack the nucleotide 71497-71652 of antisense compounds targeting SEQ ID NO:11.In certain embodiments, the short antisense compounds of targeted nucleotide 71497-71652 comprises the nucleotide sequence that is selected from SEQ ID NO 863,855,850 or 879.In some this embodiment, the short antisense compounds of the nucleotide 71497-71652 of targeting SEQ ID NO:11 is selected from Isis No 147037,147038,147039 or 147061.

In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 8-16, preferred 9-15, more preferably 9-14, more preferably 10-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 9-14 nucleotide.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 10-14 nucleotide.In certain embodiments, this short antisense compounds is short antisense oligonucleotide.

In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid is short gapmer.In some this embodiment, comprise at least one high-affinity in the short gapmer of targeting PTP1B nucleic acid one or more pterions therein and modify.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid comprises 1-3 high-affinity modification in each pterion.In some this embodiment, the nucleoside in pterion or nucleotide comprise 2 ' and modify.In some this embodiment, the monomer in pterion is BNA.In some this embodiment, the monomer in pterion is selected from α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA, β-D-methylene oxygen base (4 '-CH₂-O-2 ') BNA, ethyleneoxy group (4 '-(CH₂)₂-O-2 ') BNA, amino oxygen base (4 '-CH₂-O-N (R)-2 ') BNA and oxygen base amino (4 '-CH₂-N (R)-O-2 ') BNA.In certain embodiments, the monomer in pterion comprises in 2 ' position and is selected from following substituent group: pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) and O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.In certain embodiments, the monomer in pterion is 2 ' MOE nucleotide.

In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid comprises between 5 ' pterion and 3 ' pterion at interval.In certain embodiments, comprise 5,6,7,8,9,10,11,12,13 or 14 monomers at interval.In certain embodiments, Jian Ge monomer is not modified deoxyribonucleotide.In certain embodiments, Jian Ge monomer is not modified ribonucleotide.In certain embodiments, modify (if any) at interval and cause producing such antisense compounds, when in conjunction with its target nucleic acids, this chemical compound can be supported the cutting of RNA enzyme (including but not limited to RNA enzyme H).

In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid has the same Dan Julian key.In some this embodiment, these keys all are that thiophosphate connects key.In certain embodiments, Lian Jian is that di-phosphate ester connects key.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid has mixed matrix.

In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 8 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 9 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 10 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 11 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 12 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 13 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 14 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 15 monomers.In certain embodiments, its length of short antisense compounds of targeting PTP1B nucleic acid is 16 monomers.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid comprises 9-15 monomer.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid comprises 10-15 monomer.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid comprises 12-14 monomer.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid comprises 12-14 nucleotide or nucleoside.

In certain embodiments, the invention provides the method for the expression of regulating PTP1B.In certain embodiments, this method comprises the short antisense compounds that uses one or more targeting PTP1B nucleic acid, and wherein the short antisense compounds of this targeting PTP1B nucleic acid is about 8 to about 16, preferably 9-15, more preferably 9-14, more preferably 10-14 monomer (promptly about 8 monomer to about 16 connections).Those of ordinary skills will appreciate that, this comprises the method that the short antisense compounds that uses one or more 8,9,10,11,12,13,14,15 or 16 monomeric targeting PTP1B nucleic acid is regulated the expression of PTP1B.

In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 8 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 9 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 10 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 11 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 12 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 13 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 14 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 15 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for adjusting PTP1B comprises that using length is the short antisense compounds of 16 monomeric targeting PTP1B nucleic acid.

In certain embodiments, the method for the expression of adjusting PTP1B comprises that use comprises the short antisense compounds of 9-15 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for the expression of adjusting PTP1B comprises that use comprises the short antisense compounds of 10-15 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for the expression of adjusting PTP1B comprises that use comprises the short antisense compounds of 12-14 monomeric targeting PTP1B nucleic acid.In certain embodiments, the method for the expression of adjusting PTP1B comprises the short antisense compounds that uses the targeting PTP1B nucleic acid that comprises 12-14 nucleotide or nucleoside.

10.PTEN

In certain embodiments, the invention provides the short antisense compounds of the nucleic acid of targeting coding PTEN.In certain embodiments, this chemical compound is used for regulate the PTEN expression of cell.In some this embodiment, give animal with the short antisense compounds of targeting PTEN nucleic acid.In certain embodiments, the short antisense compounds of targeting PTEN nucleic acid can be used for studying PTEN, is used to study some nuclease and/or is used to assess the antisense activity.In some this embodiment, the short antisense compounds of targeting PTEN nucleic acid can be used for assessing some die body and/or chemical modification.In certain embodiments, the short antisense compounds of targeting PTEN nucleic acid being given animal can cause measurable phenotype to change.

The short antisense compounds of targeting PTEN can have short antisense compounds characteristic or feature any or that multiple this paper summarized.In certain embodiments, the short antisense compounds of targeting PTP1B nucleic acid has and is selected from following die body (pterion-deoxidation interval-pterion): 1-12-1,1-1-10-2,2-10-1-1,3-10-3,2-10-3,2-10-2,1-10-1,1-10-2,3-8-3,2-8-2,1-8-1,3-6-3 or 1-6-1, more preferably 1-10-1,2-10-2,3-10-3 and 1-9-2.

The short antisense compounds of some targeting PTEN nucleic acid

In certain embodiments, lacking the antisense compounds targeting has

The PTEN nucleic acid of the sequence of searching number No.NM_000314.4 (being incorporated herein) with SEQ ID NO:14.In certain embodiments, lacking the antisense compounds targeting has

The PTEN nucleic acid of the sequence of the nucleotide 8063255-8167140 of the sequence of searching number No.NT_033890.3 (being incorporated herein) with SEQ ID NO:15.In some this embodiment, short antisense compounds and the SEQ ID NO:14 of targeting SEQ ID NO:14 have at least 90% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:14 of targeting SEQ ID NO:14 have at least 95% complementarity.In some this embodiment, the short antisense compounds of targeting SEQID NO:14 and SEQ ID NO:14 have 100% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:15 of targeting SEQ ID NO:15 have at least 90% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:15 of targeting SEQ ID NO:15 have at least 95% complementarity.In some this embodiment, short antisense compounds and the SEQ ID NO:15 of targeting SEQ ID NO:15 have 100% complementarity.

In certain embodiments, the short antisense compounds of targeting SEQ ID NO:14 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 20 and 21.In certain embodiments, the short antisense compounds of targeting SEQ ID NO:15 comprises the nucleotide sequence that is selected from the nucleotide sequence that provides in table 22 and 23.

Each nucleotide sequence that provides in the table 20,21,22 and 23 is independent of any to connecting the modification of key or nuclear base between sugar moieties, nucleoside.Thus, the short antisense compounds that comprises the nucleotide sequence that provides in the table 20,21,22 and 23 can comprise one or more to connecting the modification of key or nuclear base between sugar moieties, nucleoside independently.Isis number (Isis NO.) described antisense compounds represented the nuclear base sequence with one or more to connecting key between sugar moieties, nucleoside or examining the combination of the modification of base.

Table 20 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:14.Table 22 has been enumerated the short antisense compounds that 100% complementarity is arranged with SEQ ID NO:15.The file that is marked with " gapmer die body " is represented the pterion of each short antisense compounds-at interval-pterion die body.Spacer segment comprises 2 '-deoxyribonucleotide, and each nucleotide of each pterion section comprises the sugar through 2 '-modification.The concrete sugar through 2 '-modification is also represented in " gapmer die body " file.For example, " 2-10-2MOE " means the 2-10-2gapmer die body, and wherein the spacer segment both sides of 10 2 '-deoxyribonucleotides are connected to the pterion section of 2 nucleotide, and wherein the nucleotide of pterion section is 2 '-MOE nucleotide.Connecting key between nucleoside is thiophosphate.Short antisense compounds comprises the 5-methylcytidine that substitutes not modified cytosine, unless show " not modified cytosine " in the gapmer die body file, represented in this case cytosine is not modified cytosine.For example, " 5-mC is only in the interval " expression spacer segment has the 5-methylcytidine, and the pterion section has not modified cytosine.

Nucleotide and abbreviation thereof through 2 '-modification comprise: 2 '-O-methoxy ethyl (MOE); 2 '-O-methyl (OMe); 2 '-O-(2,2,3,3,3-five fluoropropyls) (PentaF); 2 '-O-[(2-methoxyl group) ethyl]-4 '-sulfo-(2 '-MOE-4 '-sulfo-); (R)-CMOE-BNA.Shown in table 20 and 22, the pterion can include 2 ' the substituent monomer that comprises above a type.For example, 1-2-10-2MOE/PentaF/MOE represents that a nucleotide of modifying through MOE connects the interval that two nucleotide of modifying through PentaF connect 10 deoxyribonucleotides again and connects two nucleotide of modifying through PentaF again.For example, 1-1-10-22 '-(butyl acetamido)-palmitamide methylene oxygen base BNA/ methylene oxygen base BNA represents that 5 '-least significant end nucleotide is 2 '-(butyl acetamide)-palmitamide, second nucleotide is methylene oxygen base BNA nucleotide, and 3 ' pterion is methylene oxygen base BNA.Unless otherwise, otherwise cytosine is a 5-methylcytosine, and connecting key between nucleoside is thiophosphate.

Table 20: the short antisense compounds of targeting SEQ ID NO:14

ISISNo	5 ' target site	3 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQID NO
						390092	5530	5541	AGAATGAGACTT	1-10-1MOE	1514
390091	5435	5446	TGAGGCATTATC	1-10-1MOE	1522
						390090	5346	5357	AGAGTATCTGAA	1-10-1MOE	1227
390088	5162	5173	CACATTAACAGT	1-10-1MOE	1511

390087	5126	5137	GTGGCAACCACA	1-10-1MOE	1501
						390085	5031	5042	ATTTGATGCTGC	1-10-1MOE	1505
390084	4982	4993	CAAAGAATGGTG	1-10-1MOE	1215
						390082	4910	4921	AGGACTTGGGAT	1-10-1MOE	1503
390080	4833	4844	TGCTGCA CATCC	1-10-1MOE	1150
						392067	4832	4845	CTGCTGCACATCCA	Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified	1510
390078	4714	4725	CTTTCAGTCATA	1-10-1MOE	1520
						390077	4693	4704	GTCAAATTCTAT	1-10-1MOE	1252
390076	4599	4610	TTCCAATGACTA	1-10-1MOE	1506
						390075	4576	4587	GTAAGCAAGGCT	1-10-1MOE	#N/A
390074	4533	4544	ACCCTCATTCAG	1-10-1MOE	1513
						390068	4191	4202	GTAAATCCTAAG	1-10-1MOE	1515
390064	4001	4012	ACCACAGCTAGT	1-10-1MOE	1498
						390063	3977	3988	CACCAATAAGTT	1-10-1MOE	1219
390058	3828	3839	AGTAGTTGTACT	1-10-1MOE	1192
						390056	3793	3804	GGGCATATCAAA	1-10-1MOE	1521
390054	3705	3716	AACACTGCACAT	1-10-1MOE	1493
						390052	3623	3634	GACAATTTCTAC	1-10-1MOE	1492
390050	3503	3514	GTATTCAAGTAA	1-10-1MOE	1140
						390049	3479	3490	GTTAATGACATT	1-10-1MOE	1491
390047	3428	3439	TGTGTAAGGTCA	1-10-1MOE	1490
						390041	3175	3186	TTAGCACTGGCC	1-10-1MOE	1489
398076	3171	3182	CACTGGCCTTGA	1-10-1MOE	1488
						398009	3170	3183	GCACTGGCCTTGAT	2-10-2MOE	1487
398075	3111	3122	AAATCATTGTCA	1-10-1MOE	1233
						398008	3110	3123	TAAATCATTGTCAA	2-10-2MOE	1486
398074	2913	2924	GCACCAATATGC	1-10-1MOE	1248
						398007	2912	2925	AGCACCAATATGCT	2-10-2MOE	1247
398073	2681	2692	TTAGCCAACTGC	1-10-1MOE	1485
						398006	2680	2693	CTTAGCCAACTGCA	2-10-2MOE	1484
390033	2679	2690	AGCCAACTGCAA	1-10-1MOE	1483
						398072	2671	2682	GCAAACTTATCT	1-10-1MOE	1482
398005	2670	2683	TGCAAACTTATCTG	2-10-2MOE	1481
						390030	2534	2545	TTTATAAAACTG	1-10-1MOE	1074
398071	2533	2544	TTATAAAACTGG	1-10-1MOE	1480
						398004	2532	2545	TTTATAAAACTGGA	2-10-2MOE	1479
390029	2510	2521	AAAGTGCCATCT	1-10-1MOE	1478
						390028	2491	2502	TCCTAATTGAAT	1-10-1MOE	1477
398070	2481	2492	ATTTTAAATGTC	1-10-1MOE	1476
						398003	2480	2493	AATTTTAAATGTCC	2-10-2MOE	1475
390027	2455	2466	AGGTATATACAT	1-10-1MOE	1206
						398069	2451	2462	ATATACATGACA	1-10-1MOE	1474
398002	2450	2463	TATATACATGACAC	2-10-2MOE	1473
						398068	2440	2451	ACAGCTACACAA	1-10-1MOE	1472
398001	2439	2452	CACAGCTACACAAC	2-10-2MOE	1471
						390026	2438	2449	AGCTACACAACC	1-10-1MOE	1470
390025	2406	2417	GTGTCAAAACCC	1-10-1MOE	1211
						398067	2405	2416	TGTCAAAACCCT	1-10-1MOE	1210
398000	2404	2417	GTGTCAAAACCCTG	2-10-2MOE	1469
						398066	2372	2383	AGATTGGTCAGG	1-10-1MOE	1468
397999	2371	2384	AAGATTGGTCAGGA	2-10-2MOE	1467

398065	2349	2360	GTTCCTATAACT	1-10-1MOE	1466
						397998	2348	2361	TGTTCCTATAACTG	2-10-2MOE	1465
398064	2331	2342	CTGACACAATGT	1-10-1MOE	1464
						397997	2330	2343	TCTGACACAATGTC	2-10-2MOE	1463
398063	2321	2332	GTCCTATTGCCA	1-10-1MOE	1205
						397996	2320	2333	TGTCCTATTGCCAT	2-10-2MOE	1462
390022	2286	2297	CAGTTTATTCAA	1-10-1MOE	1142
						336221	2230	2243	TCAGACTTTTGTAA	3-8-3MOE	1461
336220	2224	2237	TTTTGTAATTTGTG	3-8-3MOE	1460
						336219	2209	2222	ATGCTGATCTTCAT	3-8-3MOE	1459
390021	2203	2214	CTTCATCAAAAG	1-10-1MOE	1458
						336218	2201	2214	CTTCATCAAAAGGT	3-8-3MOE	1457
389779	2201	2212	TCATCAAAAGGT	1-9-2MOE	1176
						389979	2201	2212	TCATCAAAAGGT	1-10-1MOE	1176
397995	2200	2213	TTCATCAAAAGGTT	2-10-2MOE	1456
						336217	2192	2205	AAGGTTCATTCTCT	3-8-3MOE	1455
390020	2183	2194	TCTGGATCAGAG	1-10-1MOE	1149
						336216	2182	2195	CTCTGGATCAGAGT	3-8-3MOE	1454
336215	2169	2182	TCAGTGGTGTCAGA	3-8-3MOE	1453
						398062	2166	2177	GGTGTCAGAATA	1-10-1MOE	1255
397994	2165	2178	TGGTGTCAGAATAT	2-10-2MOE	1452
						390019	2163	2174	GTCAGAATATCT	1-10-1MOE	1173
336214	2157	2170	GAATATCTATAATG	3-8-3MOE	1573
						398061	2151	2162	ATAATGATCAGG	1-10-1MOE	1451
397993	2150	2163	TATAATGATCAGGT	2-10-2MOE	1450
						336213	2146	2159	ATGATCAGGTTCAT	3-8-3MOE	1449
389778	2144	2155	TCAGGTTCATTG	1-9-2MOE	1448
						389978	2144	2155	TCAGGTTCATTG	1-10-1MOE	1448
398060	2137	2148	CATTGTCACTAA	1-10-1MOE	1447
						336212	2136	2149	TCATTGTCACTAAC	3-8-3MOE	1446
397992	2136	2149	TCATTGTCACTAAC	2-10-2MOE	1446
						336211	2112	2125	ACAGAAGTTGAACT	3-8-3MOE	1445
390017	2111	2122	GAAGTTGAACTG	1-10-1MOE	1444
						398059	2108	2119	GTTGAACTGCTA	1-10-1MOE	1443
397991	2107	2120	AGTTGAACTGCTAG	2-10-2MOE	1442
						336210	2104	2117	TGAACTGCTAGCCT	3-8-3MOE	1441
335340	2104	2118	TTGAACTGCTAGCCT	1-10-4MOE	1440
						335339	2103	2117	TGAACTGCTAGCCTC	1-10-4MOE	1439
335338	2102	2116	GAACTGCTAGCCTCT	1-10-4MOE	1438
						335337	2101	2115	AACTGCTAGCCTCTG	1-10-4MOE	1437
335336	2100	2114	ACTGCTAGCCTCTGG	1-10-4MOE	1436
						390430	2099	2111	GCTAGCCTCTGGA	The 1-10-2MOE cytosine is not modified	1163
390431	2099	2111	GCTAGCCTCTGGA	C is 9-(amino ethoxy) phenoxazine in the not modified pterion of 1-10-2MOE cytosine	1163
						390432	2099	2111	GCTAGCCTCTGGA	1-10-2MOE	1163
390433	2099	2111	GCTAGCCTCTGGA	The not modified Nt6 of 1-10-2MOE cytosine is 9-(amino ethoxy) phenoxazine	1163
						390434	2099	2111	GCTAGCCTCTGGA	1-10-2MOE	1163

				The not modified Nt 7 of cytosine is 9-(amino ethoxy) phenoxazine
						390435	2099	2111	GCTAGCCTCTGGA	The not modified Nt9 of 1-10-2MOE cytosine is 9-(amino ethoxy) phenoxazine	1163
335335	2099	2113	CTGCTAGCCTCTGGA	1-10-4MOE	1435
						389777	2098	2109	TAGCCTCTGGAT	1-9-2MOE	1434
389954	2098	2109	TAGCCTCTGGAT	1-10-1MOE	1434
						335334	2098	2112	TGCTAGCCTCTGGAT	1-10-4MOE	1433
331429	2097	2110	CTAGCCTCTGGATT	2-10-2MOE	1431
						335349	2097	2110	CTAGCCTCTGGATT	2-10-2MOE	1431
335367	2097	2110	CTAGCCTCTGGATT	2-10-2 methylene oxygen base BNA	1431
						335378	2097	2110	CTAGCCTCTGGATT	2-10-2 methylene oxygen base BNA	1431
392061	2097	2110	CTAGCCTCTGGATT	Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified	1431
						383991	2097	2109	TAGCCTCTGGATT	1-10-2 2 '-(acetylaminohydroxyphenylarsonic acid butyl-acetamido)-cholesterol/MOE	1432
383992	2097	2109	TAGCCTCTGGATT	1-10-2 2 '-(acetylaminohydroxyphenylarsonic acid butyl-acetamido)-cholic acid/MOE	1432
						386970	2097	2109	TAGCCTCTGGATT	1-10-2MOE	1432
390578	2097	2109	TAGCCTCTGGATT	T in the not modified pterion of 1-10-2MOE cytosine is the 2-thio-thymine	1432
						390614	2097	2109	TAGCCTCTGGATT	1-10-2PentaF	1432
335333	2097	2111	GCTAGCCTCTGGATT	1-10-4MOE	1430
						386683	2097	2109	TAGCCTCTGGATT	1-10-22 '-(butyl acetamido)-palmitamide/MOE	1432
371975	2096	2110	CTAGCCTCTGGATTT	3-10-2MOE	1429
						335341	2096	2111	GCTAGCCTCTGGATTT	3-10-3MOE	1428
335350	2096	2111	GCTAGCCTCTGGATTT	3-10-3MOE	1428
						335368	2096	2111	GCTAGCCTCTGGATTT	Connect key for di-phosphate ester in the 3-10-3 methylene oxygen base BNA pterion	1428
335379	2096	2111	GCTAGCCTCTGGATTT	3-10-3 methylene oxygen base BNA	1428
						383739	2096	2111	GCTAGCCTCTGGATTT	3-10-3MOE is a 5-methylcytosine in the interval	1428
384071	2096	2111	GCTAGCCTCTGGATTT	3-10-3OMe	1428

				At interval is 5-methylcytosine
						384073	2096	2111	GCTAGCCTCTGGATTT	3-10-3 methylene oxygen base BNA is a 5-methylcytosine in the interval	1428
390576	2096	2111	GCTAGCCTCTGGATTT	3-10-3MOE at interval in for the T in the 5-methylcytosine pterion be the 2-thio-thymine	1428
						390580	2096	2111	GCTAGCCTCTGGATTT	Pyrimidine in the 3-10-3MOE pterion is that the cytosine in the 5-thiazole interval is not modified	1428
390581	2096	2111	GCTAGCCTCTGGATTT	Cytosine in the 3-10-3MOE interval is not modified	1428
						391863	2096	2111	GCTAGCCTCTGGATTT	The 3-10-3MOE cytosine is not modified	1428
391864	2096	2111	GCTAGCCTCTGGATTT	Cytosine in the 3-10-3 methylene oxygen base BNA interval is not modified	1428
						391865	2096	2111	GCTAGCCTCTGGATTT	3-10-3 methylene oxygen base BNA cytosine is not modified	1428
375560	2096	2110	CTAGCCTCTGGATTT	2-10-3MOE	1429
						391172	2096	2110	CTAGCCTCTGGATTT	Methylene oxygen base BNA cytosine is not modified	1429
391175	2096	2110	CTAGCCTCTGGATTT	2-10-3 methylene oxygen base BNA	1429
						391449	2096	2110	CTAGCCTCTGGATTT	The 2-10-3MOE cytosine is not modified	1429
392054	2096	2110	CTAGCCTCTGGATTT	Cytosine in the 2-10-3 methylene oxygen base BNA interval is not modified	1429
						392055	2096	2110	CTAGCCTCTGGATTT	Cytosine in the 2-10-3MOE interval is not modified	1429
362977	2096	2111	GCTAGCCTCTGGATTT	2-12-2MOE	1428
						386770	2096	2109	TAGCCTCTGGATTT	1-11-2MOE	1427
390577	2096	2109	TAGCCTCTGGATTT	T in the not modified pterion of 1-10-3MOE cytosine is the 2-thio-thymine	1427
						335332	2096	2110	CTAGCCTCTGGATTT	1-10-4MOE	1429
390579	2096	2111	GCTAGCCTCTGGATTT	1-1-1-10-3 MOE/4 '-sulfo-	1428

				/ 2 '-O-[(2-methoxyl group) ethyl]-4 '-sulfo-/2 '-O-[(2-methoxyl group) ethyl] connect key for di-phosphate ester in the not modified pterion of cytosine in-4 '-sulfo-pterion
						391173	2096	2110	CTAGCCTCTGGATTT	2-10-3 (5 ' R)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1429
391174	2096	2110	CTAGCCTCTGGATTT	2-10-3 (5 ' S)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1429
						390607	2096	2111	GCTAGCCTCTGGATTT	Cytosine in the 3-10-3MOE/pentaF pterion is not modified	1428
390609	2096	2111	GCTAGCCTCTGGATTT	Cytosine in the 3-10-2-1 MOE/MOE/pentaF pterion is not modified	1428
						384072	2096	2111	GCTAGCCTCTGGATTT	Cytosine in the 1-2-10-3 MOE/pentaF/pentaF pterion is not modified	1428
390606	2096	2111	GCTAGCCTCTGGATTT	Cytosine in the 1-2-10-3 MOE/pentaF/pentaF pterion is not modified	1428
						390608	2096	2111	GCTAGCCTCTGGATTT	Cytosine in the 1-2-10-3 MOE/pentaF/pentaF pterion is not modified	1428
391869	2096	2111	GCTAGCCTCTGGATTT	1-2-10-3 methylene oxygen base BNA/ (5 ' S)-5 '-methyl-methylene oxygen base BNA/ (5 ' S)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1428
						385036	2096	2111	GCTAGCCTCTGGATTT	Cytosine in 1-2-10-3 OMe/2 '-O-methyl-4 '-sulfo-/2 '-O-methyl-4 '-sulfo-pterion is not modified	1428
385871	2096	2111	GCTAGCCTCTGGATTT	1-2-10-3OMe/ 2 '-O-[(2-methoxyl group) ethyl]-4 '-sulfo-/2 '-O-[(2-methoxyl group) second	1428

				Base] cytosine in-4 '-sulfo-pterion is not modified
						386682	2096	2111	GCTAGCCTCTGGATTT	1-2-10-32 '-(butyl acetamido)-palmitamide/MOE/MOE	1428
390582	2096	2111	GCTAGCCTCTGGATTT	1-2-10-3 MOE/2 '-O-[(2-methoxyl group) ethyl]-4 '-sulfo-/2 '-O-[(2-methoxyl group) ethyl] connect key for di-phosphate ester in the not modified pterion of cytosine in-4 '-sulfo-pterion	1428
						391868	2096	2111	GCTAGCCTCTGGATTT	1-2-10-3 (5 ' R)-5 '-methyl-methylene oxygen base BNA/ methylene oxygen base BNA/ (5 ' R)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1428
336209	2095	2108	AGCCTCTGGATTTG	3-8-3MOE	1425
						335331	2095	2109	TAGCCTCTGGATTTG	1-10-4MOE	1426
335376	2095	2109	TAGCCTCTGGATTTG	1-10-4 methylene oxygen base BNA	1426
						335377	2095	2109	TAGCCTCTGGATTTG	1-10-4 methylene oxygen base BNA is a di-phosphate ester in the 3 ' pterion	1426
335330	2094	2108	AGCCTCTGGATTTGA	1-10-4MOE	1424
						336208	2079	2092	GGCTCCTCTACTGT	3-8-3MOE	1423
336207	2073	2086	TCTACTGTTTTTGT	3-8-3MOE	1422
						336206	2047	2060	CACCTTAAAATTTG	3-8-3MOE	1518
389776	2046	2057	CTTAAAATTTGG	1-9-2MOE	1421
						389977	2046	2057	CTTAAAATTTGG	1-10-1MOE	1421
397990	2045	2058	CCTTAAAATTTGGA	2-10-2MOE	1420
						336205	2043	2056	TTAAAATTTGGAGA	3-8-3MOE	1419
398058	2029	2040	AGTATCGGTTGG	1-10-1MOE	1418
						336204	2028	2041	AAGTATCGGTTGGC	3-8-3MOE	1417
397989	2028	2041	AA GTATCGGTTGGC	2-10-2MOE	1417
						336203	2002	2015	TGCTTTGTCAAGAT	3-8-3MOE	1416
389775	2002	2013	CTTTGTCAAGAT	1-9-2MOE	1177
						389976	2002	2013	CTTTGTCAAGAT	1-10-1MOE	1177
397988	2001	2014	GCTTTGTCAAGATC	2-10-2MOE	1415
						336202	1959	1972	TCCTTGTCATTATC	3-8-3MOE	1414
389774	1945	1956	CACGCTCTATAC	1-9-2MOE	1413
						389975	1945	1956	CACGCTCTATAC	1-10-1MOE	1413
336201	1944	1957	GCACGCTCTATACT	3-8-3MOE	1412
						336200	1929	1942	CAAATGCTATCGAT	3-8-3MOE	1411
389773	1904	1915	AGACTTCCATTT	1-9-2MOE	1410
						389974	1904	1915	AGACTTCCATTT	1-10-1MOE	1410

336199	1902	1915	AGACTTCCATTTTC	3-8-3MOE	1409
						336198	1884	1897	TTTTCTGAGGTTTC	3-8-3MOE	1408
398057	1878	1889	GGTTTCCTCTGG	1-10-1MOE	1407
						397987	1877	1890	AGGTTTCCTCTGGT	2-10-2MOE	1406
336197	1873	1886	TTCCTCTGGTCCTG	3-8-3MOE	1405
						390015	1868	1879	GGTCCTGGTATG	1-10-1MOE	1404
398056	1865	1876	CCTGGTATGAAG	1-10-1MOE	1403
						336196	1864	1877	TCCTGGTATGAAGA	3-8-3MOE	1402
397986	1864	1877	TCCTGGTATGAAGA	2-10-2MOE	1402
						398055	1849	1860	TATTTACCCAAA	1-10-1MOE	1401
397985	1848	1861	GTATTTACCCAAAA	2-10-2MOE	1400
						336195	1847	1860	TATTTACCCAAAAG	3-8-3MOE	1399
389772	1846	1857	TTACCCAAAAGT	1-9-2MOE	1398
						389973	1846	1857	TTACCCAAAAGT	1-10-1MOE	1398
336194	1838	1851	AAAAGTGAAACATT	3-8-3MOE	1145
						398054	1836	1847	GTGAAACATTTT	1-10-1MOE	1144
397984	1835	1848	AGTGAAACATTTTG	2-10-2MOE	1397
						336193	1828	1841	CATTTTGTCCTTTT	3-8-3MOE	1182
336192	1810	1823	CATCTTGTTCTGTT	3-8-3MOE	1396
						336191	1800	1813	TGTTTGTGGAAGAA	3-8-3MOE	1395
398053	1796	1807	TGGAAGAACTCT	1-10-1MOE	1394
						397983	1795	1808	GTGGAAGAACTCTA	2-10-2MOE	1393
389771	1794	1805	GAAGAACTCTAC	1-9-2MOE	1392
						389972	1794	1805	GAAGAACTCTAC	1-10-1MOE	1392
336190	1789	1802	GAACTCTACTTTGA	3-8-3MOE	1391
						336189	1773	1786	TCACCACACACAGG	3-8-3MOE	1390
336188	1754	1767	GCTGAGGGAACTCA	3-8-3MOE	1389
						398052	1751	1762	GGGAACTCAAAG	1-10-1MOE	1388
389770	1750	1761	GGAACTCAAAGT	1-9-2MOE	1386
						389971	1750	1761	GGAACTCAAAGT	1-10-1MOE	1386
397982	1750	1763	AGGGAACTCAAAGT	2-10-2MOE	1387
						336187	1747	1760	GAACTCAAAGTACA	3-8-3MOE	1385
390012	1745	1756	TCAAAGTACATG	1-10-1MOE	1384
						336186	1688	1701	TCTTCACCTTTAGC	3-8-3MOE	1383
398051	1684	1695	CCTTTAGCTGGC	1-10-1MOE	1220
						397981	1683	1696	ACCTTTAGCTGGCA	2-10-2MOE	1382
336185	1677	1690	AGCTGGCAGACCAC	3-8-3MOE	1381
						389769	1676	1687	TGGCAGACCACA	1-9-2MOE	1249
389970	1676	1687	TGGCAGACCACA	1-10-1MOE	1249
						392060	1675	1688	CTGGCAGACCACAA	Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified	1380
398050	1672	1683	AGACCACAAACT	1-10-1MOE	1379
						397980	1671	1684	CAGACCACAAACTG	2-10-2MOE	1378
390011	1658	1669	GGATTGCAAGTT	1-10-1MOE	1238
						336184	1655	1668	GATTGCAAGTTCCG	3-8-3MOE	1508
336183	1644	1657	CCGCCACTGAACAT	3-8-3MOE	1377
						390010	1643	1654	CCACTGAACATT	1-10-1MOE	1240
398049	1641	1652	ACTGAACATTGG	1-10-1MOE	1376
						397979	1640	1653	CACTGAACATTGGA	2-10-2MOE	1375
336182	1633	1646	CATTGGAATAGTTT	3-8-3MOE	1374
						389768	1630	1641	GAATAGTTTCAA	1-9-2MOE	1373
389969	1630	1641	GAATAGTTTCAA	1-10-1MOE	1373

398048	1626	1637	AGTTTCAAACAT	1-10-1MOE	1372
						397978	1625	1638	TAGTTTCAAACATC	2-10-2MOE	1371
336181	1623	1636	GTTTCAAACATCAT	3-8-3MOE	1370
						398047	1614	1625	CATCTTGTGAAA	1-10-1MOE	1369
336180	1613	1626	TCATCTTGTGAAAC	3-8-3MOE	1368
						390009	1613	1624	ATCTTGTGAAAC	1-10-1MOE	1175
397977	1613	1626	TCATCTTGTGAAAC	2-10-2MOE	1368
						390007	1563	1574	CAGGTAGCTATA	1-10-1MOE	1367
336179	1561	1574	CAGGTAGCTATAAT	3-8-3MOE	1366
						336178	1541	1554	CATAGCGCCTCTGA	3-8-3MOE	1365
336177	1534	1547	CCTCTGACTGGGAA	3-8-3MOE	1364
						389767	1534	1545	TCTGACTGGGAA	1-9-2MOE	1151
389968	1534	1545	TCTGACTGGGAA	1-10-1MOE	1151
						335344	1503	1516	TCTCTGGTCCTTAC	2-10-2MOE	1363
335355	1503	1516	TCTCTGGTCCTTAC	Connect key for di-phosphate ester in the 2-10-2MOE pterion	1363
						335370	1503	1516	TCTCTGGTCCTTAC	Connect key for di-phosphate ester in the 2-10-2 methylene oxygen base BNA pterion	1363
335381	1503	1516	TCTCTGGTCCTTAC	2-10-2 methylene oxygen base BNA	1363
						335411	1503	1516	TCTCTGGTCCTTAC	2-10-2MOE 3 ' C is a 9-(amino ethoxy) phenoxazine	1363
335412	1503	1516	TCTCTGGTCCTTAC	C in the 2-10-2MOE 5 ' pterion is a 9-(amino ethoxy) phenoxazine	1363
						335413	1503	1516	TCTCTGGTCCTTAC	C in the 2-10-2MOE pterion is a 9-(amino ethoxy) phenoxazine	1363
336176	1502	1515	CTCTGGTCCTTACT	3-8-3MOE	1361
						335345	1502	1517	GTCTCTGGTCCTTACT	3-10-3MOE	1362
335356	1502	1517	GTCTCTGGTCCTTACT	Connect key for di-phosphate ester in the 3-10-3MOE pterion	1362
						335371	1502	1517	GTCTCTGGTCCTTACT	Connect key for di-phosphate ester in the 3-10-3 methylene oxygen base BNA pterion	1362
335382	1502	1517	GTCTCTGGTCCTTACT	3-10-3 methylene oxygen base BNA	1362
						335414	1502	1517	GTCTCTGGTCCTTACT	C in the 3-10-3MOE 3 ' pterion is a 9-(amino ethoxy) phenoxazine	1362
335415	1502	1517	GTCTCTGGTCCTTACT	C in the 3-10-3MOE 5 ' pterion is for being 9-(amino ethoxy)	1362

				Phenoxazine
						335416	1502	1517	GTCTCTGGTCCTTACT	C in the 3-10-3MOE pterion is a 9-(amino ethoxy) phenoxazine	1362
336175	1495	1508	CCTTACTTCCCCAT	3-8-3MOE	1360
						336174	1472	1485	GGGCCTCTTGTGCC	3-8-3MOE	1359
336173	1465	1478	TTGTGCCTTTAAAA	3-8-3MOE	1358
						398046	1465	1476	GTGCCTTTAAAA	1-10-1MOE	1199
389766	1464	1475	TGCCTTTAAAAA	1-9-2MOE	1217
						389967	1464	1475	TGCCTTTAAAAA	1-10-1MOE	1217
397976	1464	1477	TGTGCCTTTAAAAA	2-10-2MOE	1357
						336172	1437	1450	AATAAATATGCACA	3-8-3MOE	1356
398045	1423	1434	TCATTACACCAG	1-10-1MOE	1355
						336171	1422	1435	ATCATTACACCAGT	3-8-3MOE	1354
389765	1422	1433	CATTACACCAGT	1-9-2MOE	1353
						389966	1422	1433	CATTACACCAGT	1-10-1MOE	1353
397975	1422	1435	ATCATTACACCAGT	2-10-2MOE	1354
						390005	1400	1411	CCAGCTTTACAG	1-10-1MOE	1352
336170	1392	1405	TTACAGTGAATTGC	3-8-3MOE	1351
						398044	1382	1393	GCTGCAACATGA	1-10-1MOE	1350
336169	1381	1394	TGCTGCAACATGAT	3-8-3MOE	1349
						389764	1381	1392	CTGCAACATGAT	1-9-2MOE	1018
389965	1381	1392	CTGCAACATGAT	1-10-1MOE	1018
						397974	1381	1394	TGCTGCAACATGAT	2-10-2MOE	1349
336168	1362	1375	TCTTCACTTAGCCA	3-8-3MOE	1348
						390004	1362	1373	TTCACTTAGCCA	1-10-1MOE	1208
336167	1353	1366	AGCCATTGGTCAAG	3-8-3MOE	1347
						398043	1345	1356	CAAGATCTTCAC	1-10-1MOE	1244
336166	1344	1357	TCAAGATCTTCACA	3-8-3MOE	1346
						390003	1344	1355	AAGATCTTCACA	1-10-1MOE	1243
397973	1344	1357	TCAAGATCTTCACA	2-10-2MOE	1346
						336165	1329	1342	AAGGGTTTGATAAG	3-8-3MOE	1345
390002	1322	1333	ATAAGTTCTAGC	1-10-1MOE	1344
						336164	1318	1331	AAGTTCTAGCTGTG	3-8-3MOE	1343
398042	1305	1316	TGGGTTATGGTC	1-10-1MOE	1214
						336163	1304	1317	GTGGGTTATGGTCT	3-8-3MOE	1342
397972	1304	1317	GTGGGTTATGGTCT	2-10-2MOE	1342
						398089	1298	1309	TGGTCTTCAAAA	1-10-1MOE	1341
389763	1296	1307	GTCTTCAAAAGG	1-9-2MOE	1197
						389964	1296	1307	GTCTTCAAAAGG	1-10-1MOE	1197
398041	1294	1305	CTTCAAAAGGAT	1-10-1MOE	1196
						336162	1293	1306	TCTTCAAAAGGATA	3-8-3MOE	1340
397971	1293	1306	TCTTCAAAAGGATA	2-10-2MOE	1340
						398040	1279	1290	GTGCAACTCTGC	1-10-1MOE	1236
336161	1278	1291	TGTGCAACTCTGCA	3-8-3MOE	1235
						397970	1278	1291	TGTGCAACTCTGCA	2-10-2MOE	1235
398039	1264	1275	TAAATTTGGCGG	1-10-1MOE	1339
						397969	1263	1276	TTAAATTTGGCGGT	2-10-2MOE	1338
336160	1261	1274	AAATTTGGCGGTGT	3-8-3MOE	1337
						336159	1253	1266	CGGTGTCATAATGT	3-8-3MOE	1336
398038	1252	1263	TGTCATAATGTC	1-10-1MOE	1200
						390000	1251	1262	GTCATAATGTCT	1-10-1MOE	1194
397968	1251	1264	GTGTCATAATGTCT	2-10-2MOE	1195

336158	1227	1240	AGATTGTATATCTT	3-8-3MOE	1335
						389762	1220	1231	ATCTTGTAATGG	1-9-2MOE	1334
389963	1220	1231	ATCTTGTAATGG	1-10-1MOE	1334
						336157	1215	1228	TTGTAATGGTTTTT	3-8-3MOE	1333
336156	1202	1215	TATGCTTTGAATCC	3-8-3MOE	1332
						389998	1199	1210	TTTGAATCCAAA	1-10-1MOE	1331
397967	1198	1211	CTTTGAATCCAAAA	2-10-2MOE	1330
						336155	1190	1203	CCAAAAACCTTACT	3-8-3MOE	1500
336154	1176	1189	ACATCATCAATATT	3-8-3MOE	1329
						389761	1171	1182	CAATATTGTTCC	1-9-2MOE	1328
389962	1171	1182	CAATATTGTTCC	1-10-1MOE	1328
						398037	1170	1181	AATATTGTTCCT	1-10-1MOE	1202
397966	1169	1182	CAATATTGTTCCTG	2-10-2MOE	1327
						336153	1164	1177	TTGTTCCTGTATAC	3-8-3MOE	1326
336152	1149	1162	CCTTCAAGTCTTTC	3-8-3MOE	1325
						389996	1141	1152	TTTCTGCAGGAA	1-10-1MOE	1165
336151	1138	1151	TTCTGCAGGAAATC	3-8-3MOE	1324
						398036	1138	1149	CTGCAGGAAATC	1-10-1MOE	1323
397965	1137	1150	TCTGCAGGAAATCC	2-10-2MOE	1322
						389760	1129	1140	ATCCCATAGCAA	1-9-2MOE	1321
389961	1129	1140	ATCCCATAGCAA	1-10-1MOE	1321
						398035	1126	1137	CCATAGCAATAA	1-10-1MOE	1320
336150	1125	1138	CCCATAGCAATAAT	3-8-3MOE	1319
						397964	1125	1138	CCCATAGCAATAAT	2-10-2MOE	1319
336149	1110	1123	TTTGGATAAATATA	3-8-3MOE	1496
						389995	1106	1117	TAAATATAGGTC	1-10-1MOE	1516
336148	1100	1113	TATAGGTCAAGTCT	3-8-3MOE	1495
						398034	1099	1110	AGGTCAAGTCTA	1-10-1MOE	1300
397963	1098	1111	TAGGTCAAGTCTAA	2-10-2MOE	1494
						389994	1095	1106	CAAGTCTAAGTC	1-10-1MOE	1299
336147	1090	1103	GTCTAAGTCGAATC	3-8-3MOE	1298
						389993	1083	1094	GAATCCATCCTC	1-10-1MOE	1297
336146	1080	1093	AATCCATCCTCTTG	3-8-3MOE	1296
						398033	1077	1088	ATCCTCTTGATA	1-10-1MOE	1198
397962	1076	1089	CATCCTCTTGATAT	2-10-2MOE	1295
						336145	1070	1083	CTTGATATCTCCTT	3-8-3MOE	1294
336144	1057	1070	TTTGTTTCTGCTAA	3-8-3MOE	1293
						389759	1056	1067	GTTTCTGCTAAC	1-9-2MOE	1292
389960	1056	1067	GTTTCTGCTAAC	1-10-1MOE	1292
						392059	1055	1068	TGTTTCTGCTAACG	Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified	1291
336143	1044	1057	ACGATCTCTTTGAT	3-8-3MOE	1290
						398032	1038	1049	TTTGATGATGGC	1-10-1MOE	1222
397961	1037	1050	CTTTGATGATGGCT	2-10-2MOE	1289
						389992	1036	1047	TGATGATGGCTG	1-10-1MOE	1288
336142	1032	1045	ATGATGGCTGTCAT	3-8-3MOE	1287
						389991	1021	1032	TGTCTGGGAGCC	1-10-1MOE	1286
392058	1020	1033	ATGTCTGGGAGCCT	Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified	1285
						397960	1020	1033	ATGTCTGGGAGCCT	2-10-2MOE	1285

389990	1007	1018	TGGCTGAAGAAA	1-10-1MOE	1284
						397959	1006	1019	GTGGCTGAAGAAAA	2-10-2MOE	1283
398031	987	998	GAGAGATGGCAG	1-10-1MOE	1282
						397958	986	999	AGAGAGATGGCAGA	2-10-2MOE	1281
389758	983	994	GATGGCAGAAGC	1-9-2MOE	1280
						389959	983	994	GATGGCAGAAGC	1-10-1MOE	1280
398030	976	987	GAAGCTGCTGGT	1-10-1MOE	1143
						397957	975	988	AGAAGCTGCTGGTG	2-10-2MOE	1279
389989	953	964	TTCTGCAGGATG	1-10-1MOE	1170
						389757	941	952	GAAATGGCTCTG	1-9-2MOE	1278
389958	941	952	GAAATGGCTCTG	1-10-1MOE	1278
						397956	940	953	GGAAATGGCTCTGG	2-10-2MOE	1277
398029	931	942	TGGACTTGGCGG	1-10-1MOE	1186
						397955	930	943	CTGGACTTGGCGGT	2-10-2MOE	1276
398028	914	925	GATGCCCCTCGC	1-10-1MOE	1275
						397954	913	926	TGATGCCCCTCGCT	2-10-2MOE	1274
398027	883	894	GGACCGCAGCCG	1-10-1MOE	1155
						397953	882	895	TGGACCGCAGCCGG	2-10-2MOE	1273
389756	874	885	CCGGGTAATGGC	1-9-2MOE	1272
						389957	874	885	CCGGGTAATGGC	1-10-1MOE	1272
398026	867	878	ATGGCTGCTGCG	1-10-1MOE	1160
						397952	866	879	AATGGCTGCTGCGG	2-10-2MOE	1271
389987	848	859	CTGGATGGTTGC	1-10-1MOE	1270
						389755	806	817	AGAGGCCTGGCA	1-9-2MOE	1269
389956	806	817	AGAGGCCTGGCA	1-10-1MOE	1269
						389985	584	595	ATGGTGACAGGC	1-10-1MOE	1268
398025	581	592	GTGACAGGCGAC	1-10-1MOE	1267
						397951	580	593	GGTGACAGGCGACT	2-10-2MOE	1266
389754	312	323	TGCTCACAGGCG	1-9-2MOE	1158
						389955	312	323	TGCTCACAGGCG	1-10-1MOE	1158
398024	231	242	CAGCGGCTCAAC	1-10-1MOE	1265
						397950	230	243	ACAGCGGCTCAACT	2-10-2MOE	1264
389982	205	216	CATGGCTGCAGC	1-10-1MOE	1161
						392056	204	217	TCATGGCTGCAGCT	2-10-2 methylene oxygen base BNA	1263
394424	204	217	TCATGGCTGCAGCT	2-10-2MOE	1263
						396007	204	217	TCATGGCTGCAGCT	2-10-2 (R)-CMOE BNA cytosine is not modified	1263
396008	204	217	TCATGGCTGCAGCT	2-10-2 (S)-CMOE BNA cytosine is not modified	1263
						396009	204	217	TCATGGCTGCAGCT	2-10-2 α-L-methylene oxygen base BNA cytosine is not modified	1263
396566	204	217	TCATGGCTGCAGCT	The amino BNA cytosine of 2-10-2 oxygen base is not modified	1263
						396567	204	217	TCATGGCTGCAGCT	The amino BNA cytosine of 2-10-2N-methyl-oxygen base is not modified	1263
396568	204	217	TCATGGCTGCAGCT	2-10-2 (6R)-6-methyl methylene oxygen base BNA	1263

				Cytosine is not modified
						397913	204	217	TCATGGCTGCAGCT	Cytosine in the 2-10-2OMe interval is not modified	1263
401974	204	217	TCATGGCTGCAGCT	The 2-10-2OMe cytosine is not modified	1263
						403737	204	217	TCATGGCTGCAGCT	2-10-2 methylene oxygen base BNA is a 5-thiazole nucleus base in the pterion	1263
						404121	204	217	TCATGGCTGCAGCT	2-10-2 methylene oxygen base BNA is 5-methylcytosine a 3 ' terminal THF thiophosphate in the interval	1263
404228	204	217	TCATGGCTGCAGCT	It in the 2-10-2 methylene oxygen base BNA interval oppositely dealkalize base (5 '-terminal reverse abasic) of 5-methylcytosine 5 '-end	1263
						396024	204	217	TCATGGCTGCAGCT	2-10-2 (6 ' S)-6 '-methyl-methylene oxygen base BNA cytosine is not modified	1263
396569	204	217	TCATGGCTGCAGCT	2-10-2 (5 ' S)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1263
						396577	204	217	TCATGGCTGCAGCT	Cytosine in methylene oxygen base BNA/ methylene oxygen base BNA/2 '-(butyl acetamido)-palmitamide/interval is not modified	1263
396576	204	217	TCATGGCTGCAGCT	Cytosine in 1-1-10-22 '-(butyl acetamido)-palmitamide/methylene oxygen base BNA/ methylene oxygen base BNA interval is not modified	1263
						398023	191	202	CCGAGAGGAGAG	1-10-1MOE	1262
397949	190	203	TCCGAGAGGAGAGA	2-10-2MOE	1261
						398022	126	137	AAGAGTCCCGCC	1-10-1MOE	1260
397948	125	138	AAAGAGTCCCGCCA	2-10-2MOE	1259

Table 22: the short antisense compounds of targeting SEQ ID NO:15

ISIS No.	5 ' target site	3 ' target site	Sequence (5 '-3 ')		The Gapmer die body		SEQ ID NO
								397948	525	538	AAAGAGTCCCGCCA		2-10-2MOE		259
398022	526	537	AAGAGTCCCGCC		1-10-1MOE		260
								397949	590	603	TCCGAGAGGAGAGA		2-10-2MOE		261
398023	591	602	CCGAGAGGAGAG		1-10-1MOE		262
								394424	604	617	TCATGGCTGCAGCT		2-10-2MOE		263
397913	604	617	TCATGGCTGCAGCT		Cytosine in the 2-10-2OMe interval is not modified		1263
								401974	604	617	TCATGGCTGCAGCT		The 2-10-2Ome cytosine is not modified		1263
403737	604	617	TCATGGCTGCAGCT		2-10-2 methylene oxygen base BNA is a 5-thiazole nucleus base in the pterion		1263
								392056	604	617	TCATGGCTGCAGCT		Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified		1263
396576	604	617	TCATGGCTGCAGCT		Cytosine in 1-1-10-22 '-(butyl acetamido)-palmitamide/methylene oxygen base BNA/ methylene oxygen base BNA interval is not modified		1263
								396577	604	617	TCATGGCTGCAGCT		Cytosine in methylene oxygen base BNA/ methylene oxygen base BNA/2 '-(butyl acetamido)-palmitamide/interval is not modified		1263
404121	604	617	TCATGGCTGCAGCT		2-10-2 methylene oxygen base BNA is 5-methylcytosine a 3 ' terminal THF thiophosphate in the interval		1263
								404228	604	617	TCATGGCTGCAGCT		It in the 2-10-2 methylene oxygen base BNA interval oppositely dealkalize base of 5-methylcytosine 5 '-end		1263
396007	604	617	TCATGGCTGCAGCT		2-10-2 (R)-CMOE BNA cytosine is not modified		1263
								396008	604	617	TCATGGCTGCAGCT		2-10-2(S)-CMOE BNA		1263

				Cytosine is not modified
						396009	604	617	TCATGGCTGCAGCT	2-10-2 α-L-methylene oxygen base BNA cytosine is not modified	1263
396024	604	617	TCATGGCTGCAGCT	2-10-2 (6 ' S)-6 '-methyl-methylene oxygen base BNA cytosine is not modified	1263
						396566	604	617	TCATGGCTGCAGCT	The amino BNA cytosine of 2-10-2 oxygen base is not modified	1263
396567	604	617	TCATGGCTGCAGCT	The amino BNA cytosine of 2-10-2N-methyl-oxygen base is not modified	1263
						396568	604	617	TCATGGCTGCAGCT	2-10-2 (6R)-6-methyl methylene oxygen base BNA cytosine is not modified	1263
396569	604	617	TCATGGCTGCAGCT	2-10-2 (5 ' S)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1263
389982	605	616	CATGGCTGCAGC	1-10-1MOE	1161
						397950	630	643	ACAGCGGCTCAACT	2-10-2MOE	1264
398024	631	642	CAGCGGCTCAAC	1-10-1MOE	1265
						389955	712	723	TGCTCACAGGCG	1-10-1MOE	1158
389754	712	723	TGCTCACAGGCG	1-9-2MOE	1158
						397951	980	993	GGTGACAGGCGACT	2-10-2MOE	1266
398025	981	992	GTGACAGGCGAC	1-10-1MOE	1267
						389985	984	995	ATGGTGACAGGC	1-10-1MOE	1268
389956	1206	1217	AGAGGCCTGGCA	1-10-1MOE	1269
						389755	1206	1217	AGAGGCCTGGCA	1-9-2MOE	1269
389987	1248	1259	CTGGATGGTTGC	1-10-1MOE	1270
						397952	1266	1279	AATGGCTGCTGCGG	2-10-2MOE	1271
398026	1267	1278	ATGGCTGCTGCG	1-10-1MOE	1160
						389957	1274	1285	CCGGGTAATGGC	1-10-1MOE	1272
389756	1274	1285	CCGGGTAATGGC	1-9-2MOE	1272
						397953	1282	1295	TGGACCGCAGCCGG	2-10-2MOE	1273
398027	1283	1294	GGACCGCAGCCG	1-10-1MOE	1155
						397954	1313	1326	TGATGCCCCTCGCT	2-10-2MOE	1274
398028	1314	1325	GATGCCCCTCGC	1-10-1MOE	1275
						397955	1330	1343	CTGGACTTGGCGGT	2-10-2MOE	1276
398029	1331	1342	TGGACTTGGCGG	1-10-1MOE	1186
						397956	1340	1353	GGAAATGGCTCTGG	2-10-2MOE	1277
389958	1341	1352	GAAATGGCTCTG	1-10-1MOE	1278
						389757	1341	1352	GAAATGGCTCTG	1-9-2MOE	1278
389989	1353	1364	TTCTGCAGGATG	1-10-1MOE	1170
						397957	1375	1388	AGAAGCTGCTGGTG	2-10-2MOE	1279
398030	1376	1387	GAAGCTGCTGGT	1-10-1MOE	1143
						389959	1383	1394	GATGGCAGAAGC	1-10-1MOE	1280

389758	1383	1394	GATGGCAGAAGC	1-9-2MOE	1280
						397958	1386	1399	AGAGAGATGGCAGA	2-10-2MOE	1281
398031	1387	1398	GAGAGATGGCAG	1-10-1MOE	1282
						397959	1406	1419	GTGGCTGAAGAAAA	2-10-2MOE	1283
389990	1407	1418	TGGCTGAAGAAA	1-10-1MOE	1284
						397960	1420	1433	ATGTCTGGGAGCCT	2-10-2MOE	1285
392058	1420	1433	ATGTCTGGGAGCCT	2-10-2 methylene oxygen base BNA is a 5-methylcytosine in the pterion	1285
						389991	1421	1432	TGTCTGGGAGCC	1-10-1MOE	1286
336142	1432	1445	ATGATGGCTGTCAT	3-8-3MOE	1287
						389992	1436	1447	TGATGATGGCTG	1-10-1MOE	1288
397961	1437	1450	CTTTGATGATGGCT	2-10-2MOE	1289
						398032	1438	1449	TTTGATGATGGC	1-10-1MOE	1222
336143	1444	1457	ACGATCTCTTTGAT	3-8-3MOE	1290
						392059	1455	1468	TGTTTCTGCTAACG	2-10-2 methylene oxygen base BNA is a 5-methylcytosine in the pterion	1291
389960	1456	1467	GTTTCTGCTAAC	1-10-1MOE	1292
						389759	1456	1467	GTTTCTGCTAAC	1-9-2MOE	1292
336144	1457	1470	TTTGTTTCTGCTAA	3-8-3MOE	1293
						336145	1470	1483	CTTGATATCTCCTT	3-8-3MOE	1294
397962	1476	1489	CATCCTCTTGATAT	2-10-2MOE	1295
						398033	1477	1488	ATCCTCTTGATA	1-10-1MOE	1198
336146	1480	1493	AATCCATCCTCTTG	3-8-3MOE	1296
						389993	1483	1494	GAATCCATCCTC	1-10-1MOE	1297
336147	1490	1503	GTCTAAGTCGAATC	3-8-3MOE	1298
						389994	1495	1506	CAAGTCTAAGTC	1-10-1MOE	1299
398034	1499	1510	AGGTCAAGTCTA	1-10-1MOE	1300
						398010	1500	1513	TACAGGTCAAGTCT	2-10-2MOE	1166
398077	1501	1512	ACAGGTCAAGTC	1-10-1MOE	1167
						398011	1512	1525	CGCAGAAATGGATA	2-10-2MOE	1301
398078	1513	1524	GCAGAAATGGAT	1-10-1MOE	1302
						398012	1570	1583	TTCGCATCCGTCTA	2-10-2MOE	1303
398079	1571	1582	TCGCATCCGTCT	1-10-1MOE	1304
						398013	1663	1676	CCCTAGGTTGAATA	2-10-2MOE	1305
398080	1664	1675	CCTAGGTTGAAT	1-10-1MOE	1306
						398014	2025	2038	GTTATGCAAATCAG	2-10-2MOE	1307
398081	2026	2037	TTATGCAAATCA	1-10-1MOE	1308
						398015	2620	2633	TGACTCAGTAAATT	2-10-2MOE	1309
398082	2621	2632	GACTCAGTAAAT	1-10-1MOE	1310
						398016	2655	2668	TTAAAATTCTTGGG	2-10-2MOE	1311
398083	2656	2667	TAAAATTCTTGG	1-10-1MOE	1312
						398017	2687	2700	CCTAACTTTTAGAC	2-10-2MOE	1313
398084	2688	2699	CTAACTTTTAGA	1-10-1MOE	1314
						398018	2745	2758	ACCTGAAACTGCAA	2-10-2MOE	1315
398085	2746	2757	CCTGAAACTGCA	1-10-1MOE	1157
						398019	13166	13179	GTGTCAAAACCACT	2-10-2MOE	1316
398086	13167	13178	TGTCAAAACCAC	1-10-1MOE	1204
						398020	14675	14688	CCTATTCCCACTGA	2-10-2MOE	1317
398087	14676	14687	CTATTCCCACTG	1-10-1MOE	1318
						390033	15351	15362	AGCCAACTGCAA	1-10-1MOE	1483

398021	30985	30998	TTGGATAAATATCT	2-10-2MOE	1168
						398088	30986	30997	TGGATAAATATC	1-10-1MOE	1169
397964	31001	31014	CCCATAGCAATAAT	2-10-2MOE	1319
						336150	31001	31014	CCCATAGCAATAAT	3-8-3MOE	1319
398035	31002	31013	CCATAGCAATAA	1-10-1MOE	1320
						389961	31005	31016	ATCCCATAGCAA	1-10-1MOE	1321
389760	31005	31016	ATCCCATAGCAA	1-9-2MOE	1321
						397965	31013	31026	TCTGCAGGAAATCC	2-10-2MOE	1322
398036	31014	31025	CTGCAGGAAATC	1-10-1MOE	1323
						336151	31014	31027	TTCTGCAGGAAATC	3-8-3MOE	1324
389996	31017	31028	TTTCTGCAGGAA	1-10-1MOE	1165
						336152	31025	31038	CCTTCAAGTCTTTC	3-8-3MOE	1325
336153	31040	31053	TTGTTCCTGTATAC	3-8-3MOE	1326
						397966	31045	31058	CAATATTGTTCCTG	2-10-2MOE	1327
398037	31046	31057	AATATTGTTCCT	1-10-1MOE	1202
						389962	31047	31058	CAATATTGTTCC	1-10-1MOE	1328
389761	31047	31058	CAATATTGTTCC	1-9-2MOE	1328
						336154	31052	31065	ACATCATCAATATT	3-8-3MOE	1329
389977	31480	31491	CTTAAAATTTGG	1-10-1MOE	1421
						389776	31480	31491	CTTAAAATTTGG	1-9-2MOE	1421
397967	62446	62459	CTTTGAATCCAAAA	2-10-2MOE	1330
						389998	62447	62458	TTTGAATCCAAA	1-10-1MOE	1331
336156	62450	62463	TATGCTTTGAATCC	3-8-3MOE	1332
						336157	62463	62476	TTGTAATGGTTTTT	3-8-3MOE	1333
389963	62468	62479	ATCTTGTAATGG	1-10-1MOE	1334
						389762	62468	62479	ATCTTGTAATGG	1-9-2MOE	1334
336158	62475	62488	AGATTGTATATCTT	3-8-3MOE	1335
						390000	67987	67998	GTCATAATGTCT	1-10-1MOE	1194
397968	67987	68000	GTGTCATAATGTCT	2-10-2MOE	1195
						398038	67988	67999	TGTCATAATGTC	1-10-1MOE	1200
336159	67989	68002	CGGTGTCATAATGT	3-8-3MOE	1336
						336160	67997	68010	AAATTTGGCGGTGT	3-8-3MOE	1337
397969	67999	68012	TTAAATTTGGCGGT	2-10-2MOE	1338
						398039	68000	68011	TAAATTTGGCGG	1-10-1MOE	1339
397971	69952	69965	TCTTCAAAAGGATA	2-10-2MOE	1340
						336162	69952	69965	TCTTCAAAAGGATA	3-8-3MOE	1340
398041	69953	69964	CTTCAAAAGGAT	1-10-1MOE	1196
						389964	69955	69966	GTCTTCAAAA GG	1-10-1MOE	1197
389763	69955	69966	GTCTTCAAAAGG	1-9-2MOE	1197
						398089	69957	69968	TGGTCTTCAAAA	1-10-1MOE	1341
397972	69963	69976	GTGGGTTATGGTCT	2-10-2MOE	1342
						336163	69963	69976	GTGGGTTATGGTCT	3-8-3MOE	1342
398042	69964	69975	TGGGTTATGGTC	1-10-1MOE	1214
						336164	69977	69990	AAGTTCTAGCTGTG	3-8-3MOE	1343
390002	69981	69992	ATAAGTTCTAGC	1-10-1MOE	1344
						336165	69988	70001	AAGGGTTTGATAAG	3-8-3MOE	1345
390003	70003	70014	AAGATCTTCACA	1-10-1MOE	1243
						397973	70003	70016	TCAAGATCTTCACA	2-10-2MOE	1346
336166	70003	70016	TCAAGATCTTCACA	3-8-3MOE	1346
						398043	70004	70015	CAAGATCTTCAC	1-10-1MOE	1244
336167	70012	70025	AGCCATTGGTCAAG	3-8-3MOE	1347
						390004	70021	70032	TTCACTTAGCCA	1-10-1MOE	1208
336168	70021	70034	TCTTCACTTAGCCA	3-8-3MOE	1348
						389965	70040	70051	CTGCAACATGAT	1-10-1MOE	1018

389764	70040	70051	CTGCAACATGAT	1-9-2MOE	1018
						397974	70040	70053	TGCTGCAACATGAT	2-10-2MOE	1349
336169	70040	70053	TGCTGCAACATGAT	3-8-3MOE	1349
						398044	70041	70052	GCTGCAACATGA	1-10-1MOE	1350
336170	70051	70064	TTACAGTGAATTGC	3-8-3MOE	1351
						390005	70059	70070	CCAGCTTTACAG	1-10-1MOE	1352
389966	70081	70092	CATTACACCAGT	1-10-1MOE	1353
						389765	70081	70092	CATTACACCAGT	1-9-2MOE	1353
397975	70081	70094	ATCATTACACCAGT	2-10-2MOE	1354
						336171	70081	70094	ATCATTACACCAGT	3-8-3MOE	1354
398045	70082	70093	TCATTACACCAG	1-10-1MOE	1355
						336172	70096	70109	AATAAATATGCACA	3-8-3MOE	1356
389967	70123	70134	TGCCTTTAAAAA	1-10-1MOE	1217
						389766	70123	70134	TGCCTTTAAAAA	1-9-2MOE	1217
397976	70123	70136	TGTGCCTTTAAAAA	2-10-2MOE	1357
						398046	70124	70135	GTGCCTTTAAAA	1-10-1MOE	1199
336173	70124	70137	TTGTGCCTTTAAAA	3-8-3MOE	1358
						336174	70131	70144	GGGCCTCTTGTGCC	3-8-3MOE	1359
336175	70154	70167	CCTTACTTCCCCAT	3-8-3MOE	1360
						335345	70161	70176	GTCTCTGGTCCTTACT	3-10-3MOE	1362
335356	70161	70176	GTCTCTGGTCCTTACT	Connect key for di-phosphate ester in the 3-10-3MOE pterion	1362
						335414	70161	70176	GTCTCTGGTCCTTACT	C in the 3-10-3MOE 3 ' pterion is a 9-(amino ethoxy) phenoxazine	1362
335415	70161	70176	GTCTCTGGTCCTTACT	C in the 3-10-3MOE 5 ' pterion is a 9-(amino ethoxy) phenoxazine	1362
						335416	70161	70176	GTCTCTGGTCCTTACT	C in the 3-10-3MOE pterion is a 9-(amino ethoxy) phenoxazine	1362
336176	70161	70174	CTCTGGTCCTTACT	3-8-3MOE	1361
						335371	70161	70176	GTCTCTGGTCCTTACT	Connect key for di-phosphate ester in the 3-10-3 methylene oxygen base BNA pterion	1362
335382	70161	70176	GTCTCTGGTCCTTACT	3-10-3 methylene oxygen base BNA	1362
						335344	70162	70175	TCTCTGGTCCTTAC	2-10-2MOE	1363
335355	70162	70175	TCTCTGGTCCTTAC	Connect key for di-phosphate ester in the 2-10-2MOE pterion	1363
						335411	70162	70175	TCTCTGGTCCTTAC	2-10-2MOE 3 ' C is a 9-(amino ethoxy) phenoxazine	1363
335412	70162	70175	TCTCTGGTCCTTAC	Second C of 2-10-2MOE is 9-(amino ethoxy) phenoxazine	1363

335413	70162	70175	TCTCTGGTCCTTAC	Second of 2-10-2MOE and 3 ' terminal C are 9-(amino ethoxy) phenoxazine	1363
						335370	70162	70175	TCTCTGGTCCTTAC	Connect key for di-phosphate ester in the 2-10-2 methylene oxygen base BNA pterion	1363
335381	70162	70175	TCTCTGGTCCTTAC	2-10-2 methylene oxygen base BNA	1363
						398068	79799	79810	ACAGCTACACAA	1-10-1MOE	1472
389968	89056	89067	TCTGACTGGGAA	1-10-1MOE	1151
						389767	89056	89067	TCTGACTGGGAA	1-9-2MOE	1151
336177	89056	89069	CCTCTGACTGGGAA	3-8-3MOE	1364
						336178	89063	89076	CATAGCGCCTCTGA	3-8-3MOE	1365
336179	89083	89096	CAGGTAGCTATAAT	3-8-3MOE	1366
						390007	89085	89096	CAGGTAGCTATA	1-10-1MOE	1367
390009	89135	89146	ATCTTGTGAAAC	1-10-1MOE	1175
						397977	89135	89148	TCATCTTGTGAAAC	2-10-2MOE	1368
336180	89135	89148	TCATCTTGTGAAAC	3-8-3MOE	1368
						398047	89136	89147	CATCTTGTGAAA	1-10-1MOE	1369
336181	89145	89158	GTTTCAAACATCAT	3-8-3MOE	1370
						397978	89147	89160	TAGTTTCAAACATC	2-10-2MOE	1371
398048	89148	89159	AGTTTCAAACAT	1-10-1MOE	1372
						389969	89152	89163	GAATAGTTTCAA	1-10-1MOE	1373
389768	89152	89163	GAATAGTTTCAA	1-9-2MOE	1373
						336182	89155	89168	CATTGGAATAGTTT	3-8-3MOE	1374
397979	89162	89175	CACTGAACATTGGA	2-10-2MOE	1375
						398049	89163	89174	ACTGAACATTGG	1-10-1MOE	1376
390010	89165	89176	CCACTGAACATT	1-10-1MOE	1240
						336183	89166	89179	CCGCCACTGAACAT	3-8-3MOE	1377
397980	94786	94799	CAGACCACAAACTG	2-10-2MOE	1378
						398050	94787	94798	AGACCACAAACT	1-10-1MOE	1379
392060	94790	94803	CTGGCAGACCACAA	Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified	1380
						389970	94791	94802	TGGCAGACCACA	1-10-1MOE	1249
389769	94791	94802	TGGCAGACCACA	1-9-2MOE	1249
						336185	94792	94805	AGCTGGCAGACCAC	3-8-3MOE	1381
397981	94798	94811	ACCTTTAGCTGGCA	2-10-2MOE	1382
						398051	94799	94810	CCTTTAGCTGGC	1-10-1MOE	1220
336186	94803	94816	TCTTCACCTTTAGC	3-8-3MOE	1383
						390012	94860	94871	TCAAAGTACATG	1-10-1MOE	1384
336187	94862	94875	GAACTCAAAGTACA	3-8-3MOE	1385
						389971	94865	94876	GGAACTCAAAGT	1-10-1MOE	1386
389770	94865	94876	GGAACTCAAAGT	1-9-2MOE	1386
						397982	94865	94878	AGGGAACTCAAAGT	2-10-2MOE	1387
398052	94866	94877	GGGAACTCAAAG	1-10-1MOE	1388
						336188	94869	94882	GCTGAGGGAACTCA	3-8-3MOE	1389
336189	94888	94901	TCACCACACACAGG	3-8-3MOE	1390
						336190	94904	94917	GAACTCTACTTTGA	3-8-3MOE	1391

389972	94909	94920	GAAGAACTCTAC	1-10-1MOE	1392
						389771	94909	94920	GAAGAACTCTAC	1-9-2MOE	1392
397983	94910	94923	GTGGAAGAACTCTA	2-10-2MOE	1393
						398053	94911	94922	TGGAAGAACTCT	1-10-1MOE	1394
336191	94915	94928	TGTTTGTGGAAGAA	3-8-3MOE	1395
						336192	94925	94938	CATCTTGTTCTGTT	3-8-3MOE	1396
397984	97824	97837	AGTGAAACATTTTG	2-10-2MOE	1397
						398054	97825	97836	GTGAAACATTTT	1-10-1MOE	1144
336194	97827	97840	AAAAGTGAAACATT	3-8-3MOE	1145
						389973	97835	97846	TTACCCAAAAGT	1-10-1MOE	1398
389772	97835	97846	TTACCCAAAAGT	1-9-2MOE	1398
						336195	97836	97849	TATTTACCCAAAAG	3-8-3MOE	1399
397985	97837	97850	GTATTTACCCAAAA	2-10-2MOE	1400
						398055	97838	97849	TATTTACCCAAA	1-10-1MOE	1401
397986	97853	97866	TCCTGGTATGAAGA	2-10-2MOE	1402
						336196	97853	97866	TCCTGGTATGAAGA	3-8-3MOE	1402
398056	97854	97865	CCTGGTATGAAG	1-10-1MOE	1403
						390015	97857	97868	GGTCCTGGTATG	1-10-1MOE	1404
336197	97862	97875	TTCCTCTGGTCCTG	3-8-3MOE	1405
						397987	97866	97879	AGGTTTCCTCTGGT	2-10-2MOE	1406
398057	97867	97878	GGTTTCCTCTGG	1-10-1MOE	1407
						336198	97873	97886	TTTTCTGAGGTTTC	3-8-3MOE	1408
336199	97891	97904	AGACTTCCATTTTC	3-8-3MOE	1409
						389974	97893	97904	AGACTTCCATTT	1-10-1MOE	1410
389773	97893	97904	AGACTTCCATTT	1-9-2MOE	1410
						336200	97918	97931	CAAATGCTATCGAT	3-8-3MOE	1411
336201	97933	97946	GCACGCTCTATACT	3-8-3MOE	1412
						389975	97934	97945	CACGCTCTATAC	1-10-1MOE	1413
389774	97934	97945	CACGCTCTATAC	1-9-2MOE	1413
						336202	97948	97961	TCCTTGTCATTATC	3-8-3MOE	1414
397988	97990	98003	GCTTTGTCAAGATC	2-10-2MOE	1415
						389976	97991	98002	CTTTGTCAAGAT	1-10-1MOE	1177
389775	97991	98002	CTTTGTCAAGAT	1-9-2MOE	1177
						336203	97991	98004	TGCTTTGTCAAGAT	3-8-3MOE	1416
397989	98017	98030	AAGTATCGGTTGGC	2-10-2MOE	1417
						336204	98017	98030	AAGTATCGGTTGGC	3-8-3MOE	1417
398058	98018	98029	AGTATCGGTTGG	1-10-1MOE	1418
						336205	98032	98045	TTAAAATTTGGAGA	3-8-3MOE	1419
397990	98034	98047	CCTTAAAATTTGGA	2-10-2MOE	1420
						389977	98035	98046	CTTAAAATTTGG	1-10-1MOE	1421
389776	98035	98046	CTTAAAATTTGG	1-9-2MOE	1421
						336207	102230	102243	TCTACTGTTTTTGT	3-8-3MOE	1422
336208	102236	102249	GGCTCCTCTACTGT	3-8-3MOE	1423
						335330	102251	102265	AGCCTCTGGATTTGA	1-10-4MOE	1424
335331	102252	102266	TAGCCTCTGGATTTG	1-10-4MOE	1426
						336209	102252	102265	AGCCTCTGGATTTG	3-8-3MOE	1425
335377	102252	102266	TAGCCTCTGGATTTG	1-10-4 methylene oxygen base BNA is a di-phosphate ester in the 3 ' pterion	1426
						335376	102252	102266	TAGCCTCTGGATTTG	1-10-4 methylene oxygen base BNA	1426
390577	102253	102266	TAGCCTCTGGATTT	The 1-10-3MOE cytosine is not modified	1427

				T in the pterion is the 2-thio-thymine
						335332	102253	102267	CTAGCCTCTGGATTT	1-10-4MOE	1429
386770	102253	102266	TAGCCTCTGGATTT	1-11-2MOE	1427
						375560	102253	102267	CTAGCCTCTGGATTT	2-10-3MOE	1429
391449	102253	102267	CTAGCCTCTGGATTT	The 2-10-3MOE cytosine is not modified	1429
						392055	102253	102267	CTAGCCTCTGGATTT	Cytosine in the 2-10-3MOE interval is not modified	1429
362977	102253	102268	GCTAGCCTCTGGATTT	2-12-2MOE	1428
						371975	102253	102267	CTAGCCTCTGGATTT	3-10-2MOE	1429
386556	102253	102268	GCTAGCCTCTGGATTT	3-10-3MOE	1428
						335341	102253	102268	GCTAGCCTCTGGATTT	3-10-3MOE	1428
335350	102253	102268	GCTAGCCTCTGGATTT	3-10-3MOE	1428
						383739	102253	102268	GCTAGCCTCTGGATTT	3-10-3MOE is a 5-methylcytosine in the interval	1428
390576	102253	102268	GCTAGCCTCTGGATTT	3-10-3MOE at interval in for the T in the 5-methylcytosine pterion be the 2-thio-thymine	1428
						390580	102253	102268	GCTAGCCTCTGGATTT	Pyrimidine in the 3-10-3MOE pterion is that the cytosine in the 5-thiazole interval is not modified	1428
390581	102253	102268	GCTAGCCTCTGGATTT	Cytosine in the 3-10-3MOE interval is not modified	1428
						391096	102253	102268	GCTAGCCTCTGGATTT	3-10-3MOE	1428
391098	102253	102268	GCTAGCCTCTGGATTT	3-10-3MOE	1428
						391863	102253	102268	GCTAGCCTCTGGATTT	The 3-10-3MOE cytosine is not modified	1428
384071	102253	102268	GCTAGCCTCTGGATTT	3-10-3OMe is a 5-methylcytosine in the interval	1428
						385036	102253	102268	GCTAGCCTCTGGATTT	Cytosine in 1-2-10-3 OMe/2 '-O-methyl-4 '-sulfo-/2 '-O-methyl-4 '-sulfo-pterion is not modified	1428
335368	102253	102268	GCTAGCCTCTGGATTT	Connect key for di-phosphate ester in the 3-10-3 methylene oxygen base BNA pterion	1428
						391864	102253	102268	GCTAGCCTCTGGATTT	Cytosine in the 3-10-3 methylene oxygen base BNA interval is not	1428

				Modified
						392054	102253	102267	CTAGCCTCTGGATTT	Cytosine in the 2-10-3 methylene oxygen base BNA interval is not modified	1429
391172	102253	102267	CTAGCCTCTGGATTT	Methylene oxygen base BNA cytosine is not modified	1429
						391865	102253	102268	GCTAGCCTCTGGATTT	3-10-3 methylene oxygen base BNA cytosine is not modified	1428
391868	102253	102268	GCTAGCCTCTGGATTT	1-2-10-3 (5 ' R)-5 '-methyl-methylene oxygen base BNA/ methylene oxygen base BNA/ (5 ' R)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1428
						391869	102253	102268	GCTAGCCTCTGGATTT	1-2-10-3 methylene oxygen base BNA/ (5 ' S)-5 '-methyl-methylene oxygen base BNA/ (5 ' S)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1428
384073	102253	102268	GCTAGCCTCTGGATTT	3-10-3 methylene oxygen base BNA is a 5-methylcytosine in the interval	1428
						335379	102253	102268	GCTAGCCTCTGGATTT	3-10-3 methylene oxygen base BNA	1428
390579	102253	102268	GCTAGCCTCTGGATTT	1-1-1-10-3MOE/4 ' sulfo-/2 '-O-[(2-methoxyl group) ethyl]-4 '-sulfo-/2 '-O-[(2-methoxyl group) ethyl] connect key for di-phosphate ester in the not modified pterion of cytosine in-4 '-sulfo-pterion	1428
						390582	102253	102268	GCTAGCCTCTGGATTT	1-2-10-3MOE/4 ' sulfo-/2 '-O-[(2-methoxyl group) ethyl] connect key for di-phosphate ester in the not modified pterion of cytosine in-4 '-sulfo-pterion	1428
390606	102253	102268	GCTAGCCTCTGGATTT	Cytosine in the 1-2-10-3 MOE/pentaF/pentaF pterion is a di-phosphate ester in the not modified pterion	1428

				Lian Jian
						384072	102253	102268	GCTA GCCTCTGGATTT	Cytosine in the 1-2-10-3 MOE/pentaF/pentaF pterion is not modified	1428
385871	102253	102268	GCTAGCCTCTGGATTT	1-2-10-3OMe/ 2 '-O-[(2-methoxyl group) ethyl]-4 '-sulfo-/2 '-O-[(2-methoxyl group) ethyl] cytosine in-4 '-sulfo-pterion is not modified	1428
						390607	102253	102268	GCTAGCCTCTGGATTT	Cytosine in the 3-10-3MOE/pentaF pterion is not modified	1428
390608	102253	102268	GCTAGCCTCTGGATTT	Cytosine in the 1-2-10-3 MOE/pentaF/pentaF pterion is not modified	1428
						390609	102253	102268	GCTAGCCTCTGGATTT	Cytosine in the 3-10-2-1 MOE/MOE/pentaF pterion is not modified	1428
386682	102253	102268	GCTAGCCTCTGGATTT	1-2-10-32 '-(butyl acetamido)-palmitamide/MOE/MOE	1428
						391173	102253	102267	CTAGCCTCTGGATTT	2-10-3 (5 ' R)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1429
391174	102253	102267	CTAGCCTCTGGATTT	2-10-3 (5 ' S)-5 '-methyl-methylene oxygen base BNA cytosine is not modified	1429
						386970	102254	102266	TAGCCTCTGGATT	1-10-2MOE	1432
390578	102254	102266	TAGCCTCTGGATT	T in the not modified pterion of 1-10-2MOE cytosine is the 2-thio-thymine	1432
						335333	102254	102268	GCTAGCCTCTGGATT	1-10-4MOE	1430
331429	102254	102267	CTAGCCTCTGGATT	2-10-2MOE	1431
						335349	102254	102267	CTAGCCTCTGGATT	2-10-2MOE	1431
335367	102254	102267	CTAGCCTCTGGATT	Connect key for di-phosphate ester in the 2-10-2 methylene oxygen base BNA pterion	1431
						392061	102254	102267	CTAGCCTCTGGATT	Cytosine in the 2-10-2 methylene oxygen base BNA interval is not modified	1431
335378	102254	102267	CTAGCCTCTGGATT	2-10-2 methylene oxygen base	1431

				BNA
						383991	102254	102266	TAGCCTCTGGATT	1-10-2 2 '-(acetylaminohydroxyphenylarsonic acid butyl-acetamido)-cholesterol/MOE	1432
383992	102254	102266	TAGCCTCTGGATT	1-10-2 2 '-(acetylaminohydroxyphenylarsonic acid butyl-acetamido)-cholic acid/MOE	1432
						386683	102254	102266	TAGCCTCTGGATT	1-10-2 5 ' terminal 2 '-(butyl acetamido)-palmitamide/MOE	1432
390614	102254	102266	TAGCCTCTGGATT	1-10-2PentaF	1432
						389954	102255	102266	TAGCCTCTGGAT	1-10-1MOE	1434
335334	102255	102269	TGCTAGCCTCTGGAT	1-10-4MOE	1433
						389777	102255	102266	TAGCCTCTGGAT	1-9-2MOE	1434
390430	102256	102268	GCTAGCCTCTGGA	The 1-10-2MOE cytosine is not modified	1163
						390431	102256	102268	GCTAGCCTCTGGA	C in the not modified pterion of 1-10-2MOE cytosine is 9-(amino ethoxy) phenoxazine	1163
390432	102256	102268	GCTAGCCTCTGGA	1-10-2MOE	1163
						390433	102256	102268	GCTAGCCTCTGGA	The not modified Nt 6 of 1-10-2MOE cytosine is 9-(amino ethoxy) phenoxazine	1163
390434	102256	102268	GCTAGCCTCTGGA	The not modified Nt 7 of 1-10-2MOE cytosine is 9-(amino ethoxy) phenoxazine	1163
						390435	102256	102268	GCTAGCCTCTGGA	The not modified Nt 9 of 1-10-2MOE cytosine is 9-(amino ethoxy) phenoxazine	1163
335335	102256	102270	CTGCTAGCCTCTGGA	1-10-4MOE	1435
						335336	102257	102271	ACTGCTAGCCTCTGG	1-10-4MOE	1436
335337	102258	102272	AACTGCTAGCCTCTG	1-10-4MOE	1437
						335338	102259	102273	GAACTGCTAGCCTCT	1-10-4MOE	1438
335339	102260	102274	TGAACTGCTAGCCTC	1-10-4MOE	1439
						335340	102261	102275	TTGAACTGCTAGCCT	1-10-4MOE	1440
336210	102261	102274	TGAACTGCTAGCCT	3-8-3MOE	1441
						397991	102264	102277	AGTTGAACTGCTAG	2-10-2MOE	1442
398059	102265	102276	GTTGAACTGCTA	1-10-1MOE	1443
						390017	102268	102279	GAAGTTGAACTG	1-10-1MOE	1444
336211	102269	102282	ACAGAAGTTGAACT	3-8-3MOE	1445
						397992	102293	102306	TCATTGTCACTAAC	2-10-2MOE	1446
336212	102293	102306	TCATTGTCACTAAC	3-8-3MOE	1446
						398060	102294	102305	CATTGTCACTAA	1-10-1MOE	1447
389978	102301	102312	TCAGGTTCATTG	1-10-1MOE	1448
						389778	102301	102312	TCAGGTTCATTG	1-9-2MOE	1448
336213	102303	102316	ATGATCAGGTTCAT	3-8-3MOE	1449

397993	102307	102320	TATAATGATCAGGT	2-10-2MOE	1450
						398061	102308	102319	ATAATGATCAGG	1-10-1MOE	1451
336214	102314	102327	GAATATCTATAATG	3-8-3MOE	1139
						390019	102320	102331	GTCAGAATATCT	1-10-1MOE	1173
397994	102322	102335	TGGTGTCA GAATAT	2-10-2MOE	1452
						398062	102323	102334	GGTGTCAGAATA	1-10-1MOE	1255
336215	102326	102339	TCAGTGGTGTCAGA	3-8-3MOE	1453
						336216	102339	102352	CTCTGGATCAGAGT	3-8-3MOE	1454
390020	102340	102351	TCTGGATCAGAG	1-10-1MOE	1149
						336217	102349	102362	AAGGTTCATTCTCT	3-8-3MOE	1455
397995	102357	102370	TTCATCAAAAGGTT	2-10-2MOE	1456
						389979	102358	102369	TCATCAAAAGGT	1-10-1MOE	1176
389779	102358	102369	TCATCAAAAGGT	1-9-2MOE	1176
						336218	102358	102371	CTTCATCAAAA GGT	3-8-3MOE	1457
390021	102360	102371	CTTCATCAAAAG	1-10-1MOE	1458
						336219	102366	102379	ATGCTGATCTTCAT	3-8-3MOE	1459
336220	102381	102394	TTTTGTAATTTGTG	3-8-3MOE	1460
						336221	102387	102400	TCAGACTTTTGTAA	3-8-3MOE	1461
390022	102443	102454	CAGTTTATTCAA	1-10-1MOE	1142
						397996	102477	102490	TGTCCTATTGCCAT	2-10-2MOE	1462
398063	102478	102489	GTCCTATTGCCA	1-10-1MOE	1205
						397997	102487	102500	TCTGACACAATGTC	2-10-2MOE	1463
398064	102488	102499	CTGACACAATGT	1-10-1MOE	1464
						397998	102505	102518	TGTTCCTATAACTG	2-10-2MOE	1465
398065	102506	102517	GTTCCTATAACT	1-10-1MOE	1466
						397999	102528	102541	AAGATTGGTCAGGA	2-10-2MOE	1467
398066	102529	102540	AGATTGGTCAGG	1-10-1MOE	1468
						398000	102561	102574	GTGTCAAAACCCTG	2-10-2MOE	1469
398067	102562	102573	TGTCAAAACCCT	1-10-1MOE	1210
						390025	102563	102574	GTGTCAAAACCC	1-10-1MOE	1211
390026	102595	102606	AGCTACACAACC	1-10-1MOE	1470
						398001	102596	102609	CACAGCTACACAAC	2-10-2MOE	1471
398068	102597	102608	ACAGCTACACAA	1-10-1MOE	1472
						398002	102607	102620	TATATACATGACAC	2-10-2MOE	1473
398069	102608	102619	ATATACATGACA	1-10-1MOE	1474
						390027	102612	102623	AGGTATATACAT	1-10-1MOE	1206
398003	102637	102650	AATTTTAAATGTCC	2-10-2MOE	1475
						398070	102638	102649	ATTTTAAATGTC	1-10-1MOE	1476
390028	102648	102659	TCCTAATTGAAT	1-10-1MOE	1477
						390029	102667	102678	AAAGTGCCATCT	1-10-1MOE	1478
398004	102689	102702	TTTATAAAACTGGA	2-10-2MOE	1479
						398071	102690	102701	TTATAAAACTGG	1-10-1MOE	1480
390030	102691	102702	TTTATAAAACTG	1-10-1MOE	1074
						398005	102827	102840	TGCAAACTTATCTG	2-10-2MOE	1481
398072	102828	102839	GCAAACTTATCT	1-10-1MOE	1482
						390033	102836	102847	AGCCAACTGCAA	1-10-1MOE	1483
398006	102837	102850	CTTAGCCAACTGCA	2-10-2MOE	1484
						398073	102838	102849	TTAGCCAACTGC	1-10-1MOE	1485
398007	103069	103082	AGCACCAATATGCT	2-10-2MOE	1247
						398074	103070	103081	GCACCAATATGC	1-10-1MOE	1248
398008	103267	103280	TAAATCATTGTCAA	2-10-2MOE	1486
						398075	103268	103279	AAATCATTGTCA	1-10-1MOE	1233
398009	103327	103340	GCACTGGCCTTGAT	2-10-2MOE	1487
						398076	103328	103339	CACTGGCCTTGA	1-10-1MOE	1488

390041	103332	103343	TTAGCACTGGCC	1-10-1MOE	1489
						390047	103585	103596	TGTGTAAGGTCA	1-10-1MOE	1490
390049	103636	103647	GTTAATGACATT	1-10-1MOE	1491
						390050	103660	103671	GTATTCAAGTAA	1-10-1MOE	1140
390052	103780	103791	GACAATTTCTAC	1-10-1MOE	1492
						390054	103862	103873	AACACTGCACAT	1-10-1MOE	1493

Salt, prodrug and bioequivalence thing (bioequivalent)

Antisense compounds provided herein comprises the salt of the acceptable salt of any medicine, ester or this ester, perhaps any other functionalized chemical equivalent, this equivalent can (directly or indirectly) provide bioactive metabolites or its residue when comprising people's animal.Therefore, for example, present disclosure specification also relates to the drug acceptable salt and the other biological equivalent of the prodrug of antisense compounds and drug acceptable salt, this prodrug.

Term " prodrug " expression is with the healing potion of non-activity or the preparation of low activity form, and it is being converted to activity form (being medicine) under the effect of endogenous enzyme, chemical substance and/or condition in the middle of health or the soma.Specifically, according to disclosed method among WO 93/24510 or the WO94/26764, the prodrug version of oligonucleotide is to prepare as SATE ((S-acetyl-2-sulfur ethyl) phosphate ester) derivant.Prodrug can comprise that also one or both ends wherein comprise the antisense compounds of such nuclear base, and described nuclear base can be cut (for example connecting key by locating to mix phosphodiester backbone endways) to produce reactive compound.In certain embodiments, one or more non-drug moieties cut down to produce activity form from prodrug.In some this embodiment, this non-drug moiety is not nucleotide or oligonucleotide.

Term " drug acceptable salt " refers to can accept and the acceptable salt of medicine on the physiology of chemical compound described herein, is promptly keeping the required biological activity of parent compound not give the salt of unwanted toxicology effect again to parent compound.The sodium salt of antisense oligonucleotide is suitable for, and is widely accepted to be used for the mankind are carried out the therapeutic administration.

In certain embodiments, also provide the salt of double-strandednucleic acid (including but not limited to the dsRNA chemical compound), include but not limited to sodium salt.

G. some drugs compositions

In certain embodiments, pharmaceutical composition of the present invention comprises one or more short antisense compounds and one or more excipient.In some this embodiment, excipient is selected from water, saline solution, alcohol, Polyethylene Glycol, gelatin, lactose, amylase, magnesium stearate, Pulvis Talci, silicic acid, viscous paraffin (viscous paraffin), hydroxy methocel and polyvinylpyrrolidone.

In certain embodiments, pharmaceutical composition of the present invention prepares with known technology, and described known technology includes but not limited to mixing, dissolving, pelletize, makes dragee (dragee-making), grinds (levigating), emulsifying, encapsulation, embedding or tablet forming technique.

In certain embodiments, pharmaceutical composition of the present invention is liquid (for example suspensoid, elixir and/or a solution).In some this embodiment, composition of liquid medicine is to assign to prepare with one-tenth well known in the art, and described composition includes but not limited to water, glycerol, oil, alcohol, correctives, antiseptic and coloring agent.

In certain embodiments, pharmaceutical composition of the present invention is solid (for example tablet and/or a capsule).In some this embodiment, the solid composite medicament that comprises one or more oligonucleotide is to assign to prepare with one-tenth well known in the art, and described composition includes but not limited to starch, saccharide, diluent, granulating agent, lubricant, binding agent and disintegrating agent.

In certain embodiments, pharmaceutical composition of the present invention is to prepare as depot formulation (depotpreparation).Some this depot formulation is more more long-acting than non-depot formulation usually.In certain embodiments, this preparation is by implanting (for example subcutaneous or intramuscular) or giving by intramuscular injection.In certain embodiments, depot formulation is to prepare with suitable polymeric material or hydrophobic material (for example emulsion in acceptable oil) or ion exchange resin, perhaps is prepared into the indissoluble derivant, for example is prepared into the salt of indissoluble.

In certain embodiments, pharmaceutical composition of the present invention comprises delivery system.The example of delivery system includes but not limited to liposome and emulsion.Some delivery system can be used for preparing the some drugs compositions, comprises that those comprise the pharmaceutical composition of hydrophobic compound.In certain embodiments, use some organic solvent such as dimethyl sulfoxine.

In certain embodiments, pharmaceutical composition of the present invention comprises one or more tissue specificities and sends molecule, and the described molecule of sending is designed to one or more drug delivery of the present invention to specific tissue or cell type.For example, in certain embodiments, pharmaceutical composition comprises the liposome that is coated with tissue specificity antibody.

In certain embodiments, pharmaceutical composition of the present invention comprises the cosolvent system.Some this cosolvent system comprises for example benzylalcohol, non-polar surfactant, water miscibility organic polymer and water.In certain embodiments, this cosolvent system is used to hydrophobic compound.This cosolvent west carry can a limiting examples be VPD cosolvent system, it is the ethanol solution that comprises 3%w/v benzylalcohol, 8%w/v non-polar surfactant polysorbate Tween 80 TM and 65%w/v Liquid Macrogol.Each ratio of this cosolvent system be can change considerably, and don't the dissolubility and the toxicity characteristic of system changed.In addition, also can change each cosolvent composition itself, for example, can use other surfactants to replace the polysorbate Tween 80; Can change the classification size (fraction size) of Polyethylene Glycol; The other biological compatible polymer can replace Polyethylene Glycol, for example polyvinylpyrrolidone; Other saccharides or polysaccharide can replace dextrose.

In certain embodiments, pharmaceutical composition of the present invention comprises slow-released system.A limiting examples of this slow-released system is the semi-permeable substrate that the solid hydrophobic polymer is made.In certain embodiments, depend on its chemical characteristic, slow-released system can be at several hrs, several days, several week or some months release medicine in the time.

In certain embodiments, preparation of pharmaceutical compositions of the present invention becomes for orally give.In some this embodiment, pharmaceutical composition is to prepare by one or more oligonucleotide and one or more drug acceptable carriers are made up.Some this carrier can make pharmaceutical composition can be mixed with tablet, pill (pill), dragee (dragee), capsule, liquid agent, gel, syrup, serosity agent, suspensoid etc., for the oral absorption of experimenter.In certain embodiments, medicinal composition for oral administration obtains by oligonucleotide and one or more solid excipients are mixed.Suitable excipient includes but not limited to filler such as saccharide, comprises lactose, sucrose, mannitol or Sorbitol; Cellulosics, for example corn starch, wheaten starch, rice fecula, potato starch, gelatin, Tragacanth, methylcellulose, hydroxypropyl emthylcellulose, sodium carboxymethyl cellulose and/or polyvinylpyrrolidone (PVP).In certain embodiments, this mixture is optional is ground and the optional auxiliary substance that adds.In certain embodiments, pharmaceutical composition is formed to obtain tablet or dragee core (core).In certain embodiments, add disintegrating agent (for example crospolyvinylpyrrolidone, agar or alginic acid or its salt such as sodium alginate).

In certain embodiments, coating is provided for sugar-coat alkyl core.In some this embodiment, can use concentrated sugar solution, it can be chosen wantonly and comprise arabic gum, Pulvis Talci, polyvinylpyrrolidone, carbopol gel, Polyethylene Glycol and/or titanium dioxide, lacquer solution and appropriate organic solvent or solvent mixture.Dyestuff or pigment can be added to tablet or dragee coating.

In certain embodiments, medicinal composition for oral administration is push style (push-fit) capsule of being made by gelatin.Some this push style capsule comprises one or more medicaments of the present invention and blended with it one or more filleies such as lactose, adhesive such as starch and/or lubricant such as Pulvis Talci or magnesium stearate and optional stabilizing agent.In certain embodiments, medicinal composition for oral administration is the soft seal capsule of being made by gelatin and plasticizer such as glycerol or Sorbitol.In some soft capsule, one or more medicaments of the present invention are dissolved or be suspended in the suitable liquid, and described liquid is fatty oil (fatty oil), liquid paraffin or liquid macrogol for example.Can add stabilizing agent in addition.

In certain embodiments, preparation of pharmaceutical compositions becomes for oral administration (buccaladministration).Some this pharmaceutical composition is tablet or the lozenge of preparing in a usual manner.

In certain embodiments, preparation of pharmaceutical compositions becomes for injection to give (for example intravenous, subcutaneous, intramuscular injection etc.).In some this embodiment, pharmaceutical composition comprises carrier and preparation in liquid, aqueous, and the buffer of described liquid, aqueous for example water or physical compatibility is as Hanks solution, Ringer's solution or normal saline buffer solution.In certain embodiments, also comprise other compositions (for example help dissolved composition or serve as the composition of antiseptic).In certain embodiments, injectable suspensions is to wait with suitable liquid-carrier, suspending agent to prepare.Some medicinal composition for injections is to present with unit dosage form, for example presents in ampoule or in multi-dose container.Some medicinal composition for injections is suspensoid, solution or the Emulsion in oil-containing or water-bearing media, and can comprise blender (formulatory agent) as suspending agent, stabilizing agent and/or dispersant.Some is applicable to that the solvent of medicinal composition for injections includes but not limited to lipophilic solvent and fatty oil such as Oleum sesami, Acrawax such as ethyl oleate or triglyceride and liposome.Moisture injection suspension can comprise the material of the viscosity that can improve suspensoid, as sodium carboxymethyl cellulose, Sorbitol or glucosan.Randomly, this suspensoid also can comprise suitable stabilizers, and the dissolubility that perhaps can improve medicament is so that the material of the highly spissated solution of preparation.

In certain embodiments, preparation of pharmaceutical compositions becomes for saturating mucosa to give.In some this embodiment, the penetrating agent that will be suitable for barrier to be infiltrated is used for preparation.This penetrating agent is well known in the art.

In certain embodiments, preparation of pharmaceutical compositions becomes for sucking to give.Some this inhaled medication compositions is the form preparation with the aerosol spray (aerosol spray) in pressurized package (pressurized pack) or nebulizer (nebulizer).Some this pharmaceutical composition comprises propellant, for example dichlorodifluoromethane, Arcton 11, dichlorotetra-fluoroethane, carbon dioxide or other suitable gas.In certain embodiments, use pressurized aerosol, can determine dosage unit with the valve of transmissibility metered amount (metered amount).In certain embodiments, can prepare capsule and the medicine-storing tube (cartridge) that uses in inhaler (inhaler) or insufflator (insufflator).Some this preparation comprises medicament of the present invention and the suitable powder binder such as the mixture of powders of lactose or starch.

In certain embodiments, preparation of pharmaceutical compositions becomes for rectum to give, as suppository or enema,retention.Some this pharmaceutical composition comprises known composition, as cocoa butter and/or other glyceride.

In certain embodiments, preparation of pharmaceutical compositions becomes for topical administration.Some this pharmaceutical composition comprises gentle humidification base material, as ointment (ointment) or emulsifiable paste (cream).Exemplary suitable ointment base material includes but not limited to that vaseline, vaseline add volatile silicone, lanoline and water-in-oil emulsion such as Eucerin^TM(can be available from Beiersdorf, Cincinnati, Ohio).Exemplary suitable emulsifiable paste base material includes but not limited to Nivea Cream^TM(can be available from Beiersdorf, Cincinnati, Ohio), cold cream (cold cream, USP), Purpose Cream^TM(can be available from Johnson ﹠amp; Johnson, New Brunswick, N.J.), hydrophilic ointment (USP) and Lubriderm^TM(can be available from Pfizer, Morris Plains, N.J.).

In certain embodiments, pharmaceutical composition of the present invention comprises the oligonucleotide for the treatment of effective dose.In certain embodiments, this treatment effective dose is enough to prevent, alleviate or improves the symptom of disease or prolong the experimenter's who is treated survival period.Determining fully in those skilled in the art's limit of power of treatment effective dose.

In certain embodiments, the short antisense compounds of one or more the present invention is prepared as prodrug.In certain embodiments, when giving in vivo, prodrug is chemically converted on the biology, has more active short antisense compounds form on the medicine or in the treatment.In certain embodiments, prodrug is suitable for, because they give than corresponding activity form is easier.For example, in some cases, prodrug may have more bioavailability (for example passing through orally give) than corresponding activity form.In some cases, prodrug is compared dissolubility with corresponding activity form improvement.In certain embodiments, the water solublity of prodrug is lower than corresponding activity form.In some cases, this prodrug can pass cell membrane better, because be deleterious for mobility at water solublity in the cell membrane.In certain embodiments, prodrug is an ester.In some this embodiment, ester is hydrolyzed into carboxylic acid by metabolism when giving.In some cases, the chemical compound that contains carboxylic acid is corresponding activity form.In certain embodiments, prodrug comprises and the bonded small peptide of acid groups (polyamino acid).In some this embodiment, this peptide is cut when giving and forms corresponding activity form.

In certain embodiments, prodrug is to produce like this: pharmaceutical active compounds is modified, made can be reproduced when this reactive compound gives in vivo.Prodrug can be designed to change the metabolic stability or the transhipment feature of medicine, can cover side effect or toxicity, can improve the taste of medicine or can change other features or the characteristic of medicine.Those skilled in the art are by the knowledge of drug effect dynamic process and drug metabolism in the relevant body, if know certain pharmaceutical active compounds, the prodrug that just can design this chemical compound is (referring to for example Nogrady (1985) Medicinal Chemistry A Biochemical Approach, Oxford University Press, New York, pages 388-392).

In certain embodiments, comprise disease or the sufferer that the pharmaceutical composition of one or more medicaments of the present invention can be used for treating (particularly among the people experimenter) in the mammal.Suitable route of administration includes but not limited to following mode: in oral, internal rectum, saturating mucosa, enteral (intestinal), enteral (enteral), part, suppository, suction, the sheath, in the ventricle, intraperitoneal, intranasal, ophthalmic and parenteral (for example intravenous, intramuscular, bone marrow are interior and subcutaneous).In certain embodiments, give the interior medicine (pharmaceutical intrathecals) of sheath to realize part rather than systemic exposure.For example, can be with the pharmaceutical composition direct injection in the zone of required effect (for example at kidney or heart area).

In certain embodiments, lacking antisense compounds compares with its parent oligonucleotide and is particularly suitable for orally give.In certain embodiments, short antisense compounds is more suitable in orally give than its parent oligonucleotide, improves because they compare usefulness with its parent oligonucleotide.In certain embodiments, short antisense compounds is more suitable in orally give than its parent oligonucleotide, but has better stability availability or dissolubility property because they are compared with its parent oligonucleotide.

In yet another aspect, medicament is aseptic lyophilizing oligonucleotide, and available suitable diluent such as sterile water for injection carry out rehydration.The product of rehydration gives as subcutaneous injection liquid or as venoclysis liquid after being diluted in the saline.The oligonucleotide that freeze dried drug products comprised is to prepare with water for injection, and transfers to pH 7.0-9.0, lyophilizing then with acid or alkali in preparation process.Freeze dried oligonucleotide can be the oligonucleotide of 25-800mg.Will be appreciated that this contains 25,50,75,100,125,150,175,200,225,250,275,300,325,350,375,425,450,475,500,525,550,575,600,625,650,675,700,725,750,775 and the lyophilizing oligonucleotide of 800mg.Freeze dried drug products can be packaged in the 2mL I type clear glass bottle (handling through ammonium sulfate), clogs with the brombutyl rubber stopper, and uses aluminum FLIP-Closedtop (overseal) sealing.

Compositions of the present invention can comprise other routines in addition and be present in auxiliary element in the pharmaceutical composition, and the consumption of these compositions is levels that this area has been established.Therefore, for example, compositions can comprise other compatible pharmaceutically active material, as pruritus, astringent, local anesthetic or antiinflammatory, the material that perhaps can comprise the other various dosage forms that can be used for the physics preparation present composition is as dyestuff, correctives, antiseptic, antioxidant, opacifier, thickening agent and stabilizing agent.But this material is not answered the biological activity of each composition of the excessive interference present composition when adding.Preparation can be sterilized, and can mix with auxiliary agent if needed, described auxiliary agent is lubricant, antiseptic, stabilizing agent, wetting agent, emulsifying agent, the salt in order to influence osmotic pressure, buffer agent, coloring agent, correctives and/or aromatic substance etc. for example, and disadvantageous interaction can not take place with the oligonucleotide in the preparation these auxiliary agents.

Antisense compounds provided herein also can mix with other molecule, molecular structure or compound mixture, encapsulation, put together or otherwise associate.

This paper also describes is the pharmaceutical composition and the preparation of the antisense compounds that comprises that this paper provides.Pharmaceutical composition can give in many ways, and this depends on and need carry out topical therapeutic or whole body therapeutic and depend on the zone that will treat.In a preferred embodiment, be lung surface topical administration particularly on the surface of respiratory tract, for example spray into, suck or be blown into powder or aerosol gives by per os and/or nose.

The pharmaceutical preparation described herein that can present with unit dosage form expediently can be prepared according to the known routine techniques of medicine industry.This technology comprises makes each active component and pharmaceutical carrier or excipient that bonded step take place.In general, preparation is like this preparation: each active component and liquid-carrier, micro-solid carrier or the two are combined equably and closely, and words if necessary are to product be shaped (for example being configured as specific granular size so that send) then.In a preferred embodiment, pharmaceutical preparation is to be prepared in appropriate solvent (for example water or normal saline) for pulmonary administration, perhaps be mixed with sterile preparation, wherein carrier or other materials make the drop can form required diameter, for sending with device with inhaler, intranasal delivery device, nebulizer and other pulmonary delivery.Perhaps, pharmaceutical preparation can be mixed with dry powder, for using in Diskus.

" pharmaceutical carrier " or " excipient " can be in order to one or more delivery of nucleic acids are gone up inert media to medicine acceptable solvent, suspending agent or any other pharmacology of individuality, and they are well known in the art.Excipient can be a liquid or solid, will carry out in conjunction with the administering mode of being planned its selection, and making to provide the bulkiness and the denseness of abbreviation when other compositions of itself and nucleic acid and given pharmaceutical composition make up.

H. some treatment is used

In certain embodiments, antisense compounds is used for regulate the expression of the target gene in the animal (as the people).In certain embodiments, this chemical compound can be used to treat metabolic disease or regulates one or more disease indications.For example, described method comprises that the antisense compounds with the expression that can regulate target gene of effective dose gives described needs treatment this step with the animal of this target gene diseases associated or disease.The antisense compounds that can effectively regulate the expression of target RNA or protein expression product provided herein is considered to active antisense compounds.Active antisense compounds also comprises one or more the chemical compound that can effectively regulate in numerous disease indications, and described disease indication comprises metabolic disease or cardiovascular disease indication, and the example is described hereinafter.

The adjusting of the expression of target gene can measure in body fluid (may contain or not contain cell), tissue or the organ of this animal.Obtaining the method for sample for analysis such as body fluid (for example saliva, serum, urine), tissue (for example biopsy) or organ and prepare sample for the method for analyzing, is well known to a person skilled in the art.The analytical method of RNA and protein level is discussed hereinbefore, and is well known to a person skilled in the art.The effect of treatment can be assessed like this: by routine clinical method known in this field, measure and express relevant biological marker or disease indication with target gene from above-mentioned body fluid, tissue or organ that the animal that has contacted one or more chemical compounds described herein is collected.These biological markers include but not limited to liver transaminase, bilirubin, albumin, blood urea nitrogen, creatine and other kidneys and liver function sign; Adhesion molecule, c-proteins C reactive, chemotactic factor, cytokine and other inflammation signs in interleukin, tumor necrosis factor, the born of the same parents.

Antisense compounds provided herein can be added to suitable medicine by certain chemical compound with effective dose and can accept in the diluent or carrier and use in pharmaceutical composition.The carrier and the diluent of science and technology hands are well known to a person skilled in the art.The selection of diluent or carrier will be decided according to a plurality of factors, includes but not limited to the dissolubility and the route of administration of chemical compound.These Considerations are well known to those skilled in the art.In one aspect, antisense compounds described herein can suppress the expression of target gene.Described chemical compound also can be used for making in order to the medicine of treatment with target gene diseases associated or disease.

Also be susceptible to so that the method that body fluid, organ or tissue contact with the antisense compounds that one or more this paper provides or the compositions of effective dose.Body fluid, organ or tissue can be contacted with one or more chemical compounds, thereby cause the target gene in the cell in body fluid, the organ or tissue to express adjusted.Effective dose can be measured like this: with the method that technical staff's routine is known, monitor antisense compounds or the compositions regulating action to target nucleic acids or its product, thereby be measured to effective dose.

Give altogether

In certain embodiments, two or more antisense compounds are given altogether.In certain embodiments, pharmaceutical composition comprises antisense compounds (particularly oligonucleotide) and one or more targeting second nucleic acid target target antisense compounds of one or more targeting first nucleic acid.In these antisense compounds one or more can be short antisense compounds.In certain embodiments, pharmaceutical composition comprises two or more antisense compounds of the same nucleic acid target target of targeting zones of different.In this antisense compounds one or more can be short antisense compounds.Two or more chemical compounds of combining can use or sequential use simultaneously.

In certain embodiments, other medicament of one or more pharmaceutical compositions and one or more is given altogether.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease identical with one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament be designed to its disease for the treatment of or disease different with one or more pharmaceutical compositions of the present invention.In certain embodiments, described one or more other medicament is designed to treat the ill effect of one or more pharmaceutical compositions of the present invention.In certain embodiments, one or more pharmaceutical compositions of the present invention and another medicament are given altogether, to treat the ill effect of this another medicament.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given simultaneously.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is given at different time.In certain embodiments, other medicament of one or more pharmaceutical compositions of the present invention and one or more is prepared together in single dosage form.In certain embodiments, with other separately preparation of medicament of one or more pharmaceutical compositions of the present invention and one or more.

In certain embodiments, can comprise that lipid reduces medicament with the medicament that pharmaceutical composition of the present invention gives altogether.In some this embodiment, can include but not limited to atorvastatin, simvastatin, rosuvastatin with the medicament that pharmaceutical composition of the present invention gives altogether and according to Ezetimibe.In some this embodiment, lipid is reduced medicament give before the pharmaceutical composition of the present invention giving.In some this embodiment, lipid is reduced medicament after giving pharmaceutical composition of the present invention, give.In some this embodiment, lipid is reduced medicament when giving pharmaceutical composition of the present invention, give.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is identical with the dosage that gives separately to be given when lipid reduces medicament.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is lower than the dosage that is given when giving lipid reduction medicament separately.In some this embodiment, the dosage that the lipid that gives altogether reduces medicament is higher than the dosage that is given when giving lipid reduction medicament separately.

In certain embodiments, it is the HMG-CoA reductase inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, the HMG-CoA reductase inhibitor is an inhibin.In some this embodiment, inhibin is selected from atorvastatin, simvastatin, pravastatin, fluvastatin and rosuvastatin.In certain embodiments, it is cholesterol absorption inhibitor that the lipid that gives altogether reduces medicament.In some this embodiment, cholesterol absorption inhibitor is according to Ezetimibe.In certain embodiments, it is HMG-CoA reductase inhibitor and cholesterol absorption inhibitor formulated together altogether that the lipid that gives altogether reduces medicament.In some this embodiment, it is according to Ezetimibe/simvastatin that lipid formulated together altogether reduces medicament.In certain embodiments, it is microsomal triglyceride transfer protein inhibitor (MTP inhibitor) that the lipid that gives altogether reduces medicament.

In certain embodiments, the medicament that gives altogether is a bile acid chelating agent.In some this embodiment, bile acid chelating agent is selected from colestyramine, colestipol and colesevelam.

In certain embodiments, the medicament that gives altogether is a nicotinic acid.In some this embodiment, nicotinic acid is selected from rapid release nicotinic acid, extended release nicotinic acid and slow release nicotinic acid.

In certain embodiments, the medicament that gives altogether is a Carboxymethylcellulose.In some this embodiment, Carboxymethylcellulose is selected from gemfibrozil, fenofibrate, chlorine Bei Te, bezafibrate and ciprofibrate.

More examples of the medicament that can give altogether with pharmaceutical composition of the present invention include but not limited to corticosteroid (including but not limited to prednisone); Immunoglobulin (includes but not limited to vein immunoglobulin (IVIg); Analgesic (for example acetaminophen); Antiinflammatory (including but not limited to NSAID (non-steroidal anti-inflammatory drug) (for example ibuprofen, COX-1 inhibitor and cox 2 inhibitor)); Salicylate; Antibiotic; Antiviral agent; Antifungal; Antidiabetic medicine (for example biguanide, glucosidase inhibitor, insulin, sulfonylureas and thiazolidinediones); The adrenergic modulation agent; Diuretic; Hormone (for example short anabolism steroid, androgen, estrogen, calcitonin, progesterone, somatostatin and thyroxin); Immunomodulator; Muscle relaxant; Hydryllin; Osteoporosis medicament (for example diphosphonate, calcitonin and estrogen); Prostaglandin; Antineoplastic agent; The spiritual healing medicament; Tranquilizer; Poison Oak Tree (oak) or black poison wood (sumac) product; Antibody and vaccine.

In certain embodiments, pharmaceutical composition of the present invention can be reduced therapy in conjunction with lipid gives.In some this embodiment, it is that curative life style changes that lipid reduces therapy.In some this embodiment, it is the LDL Apheresis that lipid reduces therapy.

I. test kit, research reagent and diagnosis

Antisense compounds provided herein can be used for diagnosis and uses as research reagent and test kit (kit).In addition, can specific inhibition of gene expression or the regulator gene antisense compounds of expressing, often be used for explaining the function of specific gene by the technical staff, perhaps be used for distinguishing each member's of certain biological approach function.

For the use in test kit and diagnosis, antisense compounds as herein described can be combined individually or with other chemical compounds or healing potion, as the instrument of difference and/or fabric analysis, with explaination cell and certain part of the central gene of expressing of tissue or the expression pattern of whole complementary series (entire complement).The method of gene expression analysis is well known to a person skilled in the art.

J. lack some advantage of antisense compounds

In certain embodiments, lacking antisense compounds compares with its parent oligonucleotide and has advantage.For example, in certain embodiments, short antisense compounds to the affinity of target nucleic acids greater than its parent oligonucleotide.In certain embodiments, the external usefulness of lacking antisense compounds is higher than its parent oligonucleotide.In some this embodiment, the external usefulness of this raising can not be explained by the affinity that improves fully.In certain embodiments, the external usefulness of this raising is attributable to that short antisense compounds infiltrates the raising of the ability in the cell and/or near the raising of the ability of the target nucleic acids in the cell.In certain embodiments, lack that usefulness is higher than its parent oligonucleotide in the body of antisense compounds.In certain embodiments, the interior usefulness of this higher body is not owing to the raising of external usefulness or the improve of affinity.In certain embodiments, short antisense compounds is compared with its parent oligonucleotide, and usefulness is much larger than according to the external usefulness or the usefulness that can predict according to affinity in the body that it had.In certain embodiments, in the body of this raising usefulness be attributable to the raising of bioavailability, cytotropic infiltration better, in case enter behind the cell to target nucleic acids near better or other factors.

In certain embodiments, can expect that the specificity that short antisense compounds and its parent oligonucleotide compare its target nucleic acids is lower.In some this embodiment, can expect that short antisense compounds can cause side effect to increase, comprise that the potential of toxic action may.In certain embodiments, do not observe this other side effect.In certain embodiments, the specific combinative non-target nucleic acids of short antisense compounds can not can touch for short antisense compounds.In this embodiment, comprise side-effect problem the lacking of toxicity than expection.

In certain embodiments, because short antisense compounds is less, their unlikely conjugated proteins.In some this embodiment, this less protein bound causes toxicity lower, because protein bound can cause undesired consequence.In certain embodiments, this less protein bound causes usefulness higher, has more antisense compounds can supply the performance therapeutical effect because this makes.In certain embodiments, less protein bound causes the toxic reduction of drug-drug interactions.

Non-limiting disclosure and being attached to herein by reference

In conjunction with some embodiment some chemical compound as herein described, compositions and method are specifically described, but following examples are only in order to illustrating chemical compound as herein described, and are not intended to and limit these chemical compounds.The list of references of quoting among the application, GenBank searching number etc., its each all by reference integral body be attached to herein.

Embodiment 1

Cell culture and handle with short antisense compounds

The effect that short antisense compounds is expressed target nucleic acids can be tested in any of the cell line in multiple cultivation or former generation.Cell line can be available from the source that can openly obtain, as American Type Culture Collection (ATCC, Manassas, VA).Cell is cultivated according to method known to a person of ordinary skill in the art.

When cell reached suitable degree of being paved with, by specification used

Handle cell with oligonucleotide.When cell reaches 65-75% degree of being paved with, handle cell with oligonucleotide.With oligonucleotide with

Invitrogen Life Technologies, Carlsbad is CA) at Opti-

(Invitrogen LifeTechnologies, Carlsbad mix in CA) low blood serum medium, to reach required oligonucleotide concentration and 2.5 or 3 μ g/mL/100nM oligonucleotide

Concentration.With this transfection mixture about 0.5 hour of incubation at room temperature.For the cell of in 96 orifice plates, growing, with each hole with 100 μ L OPTI-

Washing is once handled with the transfection mixture of 130 μ L then.For the cell of in the tissue culturing plate of 24 orifice plates or other standards, growing, carry out similar processing with the culture medium and the oligonucleotide of proper volume.Repeating twice or three times handles cell and collects data.After handling about 4-7 hour under 37 ℃, contain the culture medium of transfection mixture with the fresh culture displacement.Harvesting after oligonucleotide is handled 16-24 hour.

The use control oligonucleotide is measured the best oligomeric compounds concentration for specific cells system.In addition, when test in oligomeric compounds screening experiment or phenotype test during oligomeric compounds, also parallel testing control oligonucleotide.

Used oligonucleotide concentration is different with the difference of cell line.For measuring the best oligonucleotide concentration for specific cells system, handle cell with the positive control oligonucleotide of a plurality of concentration.The positive control oligonucleotide concentration that will cause 80% of target mRNA to suppress then, as in the subsequent experimental of this cell line to the screening concentration of new oligonucleotide.Do not suppress if reach 80%, then will cause the 60% positive control oligonucleotide least concentration that suppresses of target mRNA, as the screening of the oligonucleotide in the subsequent experimental of this cell line concentration.Do not suppress if reach 60%, think that then this specific cells system is not suitable for the oligonucleotide transfection experiment.The concentration of antisense oligonucleotide used herein is when this antisense oligonucleotide is 50nM-300nM during with the liposome reagent transfection, when antisense oligonucleotide is 1nM-40nM during by electroporation transfection.

Embodiment 2: the real-time quantitative PCR analysis of target mRNA level

Target mRNA level quantitatively be to use ABI

7600,7700 or 7900Sequence Detection System (PE-Applied Biosystems, Foster City CA) finish by real-time quantitative PCR according to manufacturer's description.

Before carrying out quantitative PCR, assessment earlier has the ability of specific primer-probe groups and GAPDH amplified reaction " multiple carry out (multiplexed) " to measured target gene.Carry out in regular turn reverse transcription (RT) reaction and PCR in real time after RNA separates, two reactions are all carried out in identical hole.RT and PCR reagent available from Invitrogen Life Technologies (Carlsbad, CA).RT, PCR in real time are in identical hole, by with 20 μ L PCR mixture (cocktail) (2.5x PCR buffer (decrement MgCl₂) (buffer minus MgCl₂), 6.6mM MgCl₂, dATP, dCTP, dCTP and the dGTP of each 375 μ M, the forward primer of each 375nM and reverse primer, the probe of 125nM, 4 RNA of unit enzyme inhibitors, 1.25 units

Taq, 5 MuLV of unit reverse transcription and 2.5xROX dyestuff) be added to and carry out in 96 orifice plates that contain 30 μ L total rna solutions (20-200ng).The RT reaction is to carry out in 30 minutes at 48 ℃ of following incubations.Activated in 10 minutes at 95 ℃ of following incubations

Behind the Taq, carry out 40 cyclic two step PCR schemes: 95 ℃, 15 seconds (degeneration), 60 ℃ then, 1.5 minutes (annealing/extension).

The gene target quantity that obtains by RT, PCR in real time is with following dual mode normalization: use the expression normalization of GAPDH (a kind of its express constant gene), perhaps by usefulness

(OR) quantitatively total RNA comes normalization for Molecular Probes, Inc.Eugene.It is by moving simultaneously with target, multiplely carry out (multiplexing) or carry out (separately) separately, coming to be undertaken quantitative by RT, PCR in real time that GAPDH expresses.Total RNA uses

(Molecular Probes, Inc.Eugene OR) carry out quantitatively the RNA quantitative reagent.

With 170 μ L's

Work reagent (

Reagent is diluted in 10mM Tris-HCl at 1: 350,1mM EDTA, pH 7.5) pipette in 96 orifice plates of the cell RNA that contains 30 μ L purification.This plate exists

Carry out reading with 485nm excitation wavelength and 530nm emission wavelength among 4000 (the PE Applied Biosystems).

GAPDH PCR probe have with 5 ' terminal covalently bound JOE and with 3 ' terminal covalently bound TAMRA or MGB, wherein JOE is a fluorescence report dyestuff, TAMRA or MGB are the quencher dyestuffs.In some cell types, use to measuring GAPDH and express from the primer of the GAPDH sequential design of different plant species and probe.For example, personnel selection GAPDH primer and probe groups measurement are spread out from cell and the expression of the GAPDH in the cell line of monkey.

Probe that PCR in real time is used and primer are designed to use conventional method and target nucleic acids hybridization.For example conventionally use

(CA) software designs probe and the primer that PCR in real time is used for Applied Biosystems, Foster City.The example of the target nucleic acids that hybridize primer and probe sequence and they provides in table 24.Target specific PCR probe have with 5 ' terminal covalently bound FAM and with 3 ' terminal covalently bound TAMRA or MGB, wherein FAM is a fluorescent dye, TAMRA or MGB are the quencher dyestuffs.

Table 24

The target Auele Specific Primer and the probe that are used for PCR in real time

The target title	Species	Sequence type	Sequence (5 '-3 ')	SEQID NO
					ApoB	Mice	Forward primer	CGTGGGCTCCAGCATTCTA	1524
ApoB	Mice	Reverse primer	AGTCATTTCTGCCTTTGCGTC	1525
					ApoB	Mice	Probe	CCAATGGTCGGGCACTGCTCAA	1526
ApoB	Mice	Forward primer	GAAAATAGACTTCCTGAATAACTATGCATT	1527
					ApoB	Mice	Reverse primer	ACTCGCTTGCCAGCTTGC	1528
ApoB	Mice	Probe	TTTCTGAGTCCCCGTGCCCAACA	1529
					GCGR	Mice	Forward primer	TGAGCCTTGCCACCTTCTCT	1530
GCGR	Mice	Reverse primer	GCGCACCCCAGCCAA	1531
					GCGR	Mice	Probe	AGAGGAGCTTCTTTTCCCTCTACCTGGGC	1532
GCGR	Mice	Forward primer	ATTTCCTGCCCCTGGTACCT	1533

The target title	Species	Sequence type	Sequence (5 '-3 ')	SEQ ID NO
					GCGR	Mice	Reverse primer	CGGGCCCACACCTCTTG	1534
GCGR	Mice	Probe	CCACAAAGTGCAGCACCGCCTAGTGT	1535
					PTEN	Mice	Forward primer	GCCACAGGCTCCCAGACAT	1536
PTEN	Mice	Reverse primer	TCCATCCTCTTGATATCTCCTTTTG	1537
					PTEN	Mice	Probe	ACAGCCATCATCAAAGAGATCGTTAGCAGAA	1538
PTEN	Mice	Forward primer	ATGACAATCATGTTGCAGCAATTC	1539
					PTEN	Mice	Reverse primer	CGATGCAATAAATATGCACAAATCA	1540
PTEN	Mice	Probe	CTGTAAAGCTGGAAAGGGACGGACTGGT	1541

Embodiment 3: targeting ApoB nucleic acid and have 2 '-MOE or methylene oxygen base (4 '-CH₂-O-2 ') the short antisense compounds of BNA modification

Give six ages in week male Balb/c mice (Jackson Laboratory, Bar Harbor, the ME) antisense compounds of peritoneal injection (i.p.) targeting ApoB, weekly twice, continuous three weeks.Antisense compounds dosage comprises 2.4,1.2,0.6,0.3 and 0.15 μ mol/kg.For the antisense compounds of 14 nucleotide of length, these dosage equal about 12,6,3,1.5 or 0.75mg/kg respectively.Shown in the table 25 is the sequence and the die body of antisense compounds used in this research.The length of these antisense compounds is 20 or 14 nucleotide, has central authorities' " at interval " zone that 10 2 '-deoxyribonucleotides are formed, and the interval region both sides are connected to 2 '-O-methoxy ethyl (" pterion " that 2 '-MOE) the wing or BNA modify.For example, 2-10-2MOE gapmer die body represents to have the interval of 10 nucleotide, both sides are connected to the antisense compounds in the 2 nucleotide pterions of modifying with 2 '-MOE at interval.The black matrix residue is represented 2 '-O-methoxy ethyl part, and the italic residue is represented methylene oxygen base (4 '-CH₂-O-2 ') BNAs.Connecting key between the nucleoside of each chemical compound all is thiophosphate.All cytosine residues of ISIS 147764 and ISIS 372938 are all replaced by 5-methylcytosine.For ISIS 387462, only there is the cytosine residue in the chemical compound pterion to be replaced by 5-methylcytosine.The ApoB-100 sequence that ApoB antisense compounds targeting can openly obtain comprises Genbank searching number XM_137955.5 (SEQ ID NO:2).

Table 25: the antisense compounds of targeting ApoB nucleic acid

ISIS NO	Target SEQ ID NO	5 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						147764	2	8865	GTCCCTGAAGATGTCAATGC	5-10-5MOE	1561

ISISNO	Target SEQ ID NO	5 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						372938	2	8235	GGTACATGGAAGTC	2-10-2MOE	190
387462	2	8235	GGTACATGGAAGTC	2-10-2 methylene oxygen base (4 '-CH2-O-2’) BNA	190

Inject the last time and put to death mice in back 48 hours, with assessment transaminase's (table 26); Liver and kidney weight (table 27); Triglyceride, LDL, HDL and free fatty acid levels (table 28); Target mRNA level (table 29) in the liver; Target proteins matter level in the blood plasma; With oligonucleotide tissue concentration (table 30).These terminal points (endpoint) are to measure with method described herein and known to a person of ordinary skill in the art.

Table 26:ALT and AST level (IU/L)

ISIS NO	Dosage μ mol/kg	ALT	AST
				Saline	N/A	27.8	46.3
147764	2.4	29.5	64.0
				372938	2.4	26.0	49.0
372938	1.2	24.8	49.5
				372938	0.6	28.0	79.3
372938	0.3	28.3	60.0
				372938	0.15	28.3	50.3
387462	2.4	41.3	84.0
				387462	1.2	35.3	63.5
387462	0.6	32.0	77.3
				387462	0.3	27.8	55.0
387462	0.15	29.3	68.3

Table 27: liver and kidney weight (accounting for the percent of saline control)

ISIS NO	Dosage μ mol/kg	Liver	Kidney
				Saline	N/A	100	100
147764	2.4	102	105
				372938	2.4	100	100
372938	1.2	90	101
				372938	0.6	96	112

ISIS NO	Dosage μ mol/kg	Liver	Kidney
				372938	0.3	91	107
372938	0.15	96	98
				387462	2.4	116	90
387462	1.2	113	90
				387462	0.6	106	97
387462	0.3	101	126
				387462	0.15	95	100

Handle between the animal saline treatment animal or oligonucleotide, TBW and food consumption quantity do not have significant difference.Glucose level between all processed group is also similar.

Table 28: triglyceride (TRIG), T-CHOL (CHOL), HDL, LDL and free fatty (FFA) level

ISIS NO	Dosage μ mol/kg	TRIG (mg/dL)	CHOL (mg/dL)	HDL (mg/dL)	LDL (mg/dL)	FFA (mg/dL)
							Saline	N/A	167	107	81.8	11.0	1.76
147764	2.4	167	107	81.3	10.3	1.29
							372938	2.4	153	104	79.0	10.3	1.28
372938	1.2	136	101	77.8	9.5	1.70
							372938	0.6	184	110	83.3	10.8	1.66
372938	0.3	138	109	84.3	11.0	1.53
							372938	0.15	151	106	82.8	10.8	1.57
387462	2.4	49	14	9.0	1.5	0.74
							387462	1.2	71	23	16.5	2.0	0.76
387462	0.6	150	55	39.3	3.7	1.43
							387462	0.3	136	92	72.8	7.5	1.14
387462	0.15	163	104	81.5	9.3	1.47

Table 29:%ApoB mRNA level (with respect to saline control)

ISIS NO	2.4 μmol/kg	1.2 μmol/kg	0.6 μmol/kg	0.3 μmol/kg	0.15 μmol/kg
						147764	57.7	ND	ND	ND	ND
372938	77.0	90.0	87.3	92.6	93.1
						387462	1.5	8.5	27.4	58.9	75.8

Caused the remarkable and dose dependent of triglyceride, T-CHOL, HDL, LDL and free fatty to descend with ISIS 387462 processing.Find according to these phenotypes, handle the ApoB mRNA (table 29) that also caused in the mice plasma and the dose dependent decline of protein (not shown) level with ISIS387462.For determining at methylene oxygen base (4 '-CH₂-O-2 ') whether the viewed efficient raising of BNA gapmer is because due to the increase of oligonucleotide accumulation, has measured total length and total oligonucleotide concentration in liver and the kidney.

Table 30: total length and total antisense compounds tissue concentration (μ M), with respect to ApoB mRNA level

(accounting for the percent of saline saline control)

ISIS NO	Dosage μ mol/kg	The kidney total length	The liver total length	Kidney is total	Liver is total	ApoB mRNA
							147764	2.4	28.6	22.9	33.5	31.3	58
372938	2.4	32.0	5.49	34.0	7.76	77
							387462	2.4	37.2	5.69	38.9	7.31	1.5
387462	1.2	29.8	3.71	31.3	4.91	8.5
							387462	0.6	18.9	1.97	20.0	2.57	27
387462	0.3	9.11	0.73	9.49	0.78	59
							387462	0.15	6.97	0.19	7.43	0.24	76

The level of 2-10-2 methylene oxygen base (4 '-CH2-O-2 ') BNA gapmer is similar with 2-10-2 MOE gapmer to 5-10-5 in kidney, but significantly reduces in liver.The EC of ISIS387462 in liver₅₀Compare to determine by inhibition with oligonucleotide concentration in the liver and ApoBmRNA.The EC of ISIS 387462₅₀Be approximately 1 μ M.Contrast therewith, the effectively EC of 5-10-5MOE gapmer chemical compound in liver₅₀Be generally 15 μ M.

These results prove to have methylene oxygen base (4 '-CH generally in the pterion₂-O-2 ') the short gapmer of ApoB is the potent inhibitor that target mRNA expresses, effectively triglyceride reducing, cholesterol and free fatty.As if the usefulness of short antisense compounds is not the result that tissue accumulation increases, because with respect to 5-10-5 MOE gapmer, observing this chemical compound has similar level in kidney, the level in liver lowers.In addition, methylene oxygen base (4 '-CH₂-O-2 ') BNA gapmer seldom shows and even does not show adverse side effect.

Embodiment 4: targeting GCGR nucleic acid and have the short antisense compounds that 2 '-MOE modifies

By peritoneal injection give eight age in week the C57/BL6 mice (Jackson Laboratory, Bar Harbor, ME) the GCGR oligonucleotide of single dose, concentration is 6.25,12.5,25 or 50mg.Each dosage group has four mices.Shown in the table 31 is sequence, die body and the conjugate that is used for the GCGR antisense compounds of this research.The runic residue is represented 2 '-O-methoxy ethyl (2 '-MOE) part.All chemical compounds all are to comprise between the thiophosphate nucleoside to connect key without exception, and each cytosine is replaced by 5-methylcytosine.ISIS 386626, ISIS 386627 and ISIS 386628 also comprise by diamides and connect key (2 '-OCH₂C (=O) N (H) (CH₂)₄N (H) C (=O)-(CH₂)₁₅CH₃) with the C that is connected of 2 '-O position of sugar₁₆Conjugated group.The GCGR sequence that GCGR antisense compounds energy targeting has been announced comprises

Searching number BC031885.1 (SEQ ID NO:7) is interior.

Table 31: the short antisense compounds of targeting GCGR nucleic acid

ISIS NO	Target SEQ ID NO	5 ' target site	Sequence (5 '-3 ')	The Gapmer die body	Conjugate	SEQ ID NO
							148364	7	393	TGCACTTTGTGGTACCAAGG	5-10-5 MOE	Do not have	1562
386626	7	1768	GC16CTTCTCCATCATA	2-10-2 MOE	C16	1563
							386627	7	1244	GC16GGCATGCTCGTCA	2-10-2 MOE	C16	653
386593	7	1244	GGGCATGCTCGTCA	2-10-2 MOE	Do not have	649
							386628	7	1680	TC16GTCTTGCTGCTTT	2-10-2 MOE	C16	1564
386594	7	1680	TGTCTTGCTGCTTT	2-10-2 MOE	Do not have	1565

Put to death mice in back 48 hours in injection, to measure serum transaminase level (table 32); Liver white adipose tissue (WAT), spleen and kidney weight (table 33); Triglyceride and glucose level (table 34); GCGR mRNA level (table 35-41); And the total length in liver and the kidney and total oligonucleotide concentration (table 42).These terminal points are to assess with method described herein and known to a person of ordinary skill in the art.Include the data of handling preceding blood-letting (Pre-Bleed) and handling back blood-letting (Post-Bleed) in.

Table 32:ALT﹠amp; AST level (IU/L)

ISIS NO	Dosage (mg/kg)	ALT Pre-Bleed	ALT Post-Bleed	AST Pre-Bleed	AST Post-Bleed
						Saline	N/A	36	51	55	85
148364	50	24	40	40	115
						148364	25	26	35	42	87
148364	12.5	23	32	44	69
						148364	6.25	28	34	47	76
386626	50	28	40	48	120
						386626	25	30	36	44	92
386626	12.5	28	34	44	90
						386626	6.25	26	42	46	69
386627	50	27	457	42	451
						386627	25	29	97	45	142
386627	12.5	29	62	46	81
						386627	6.25	23	87	38	96
386593	50	23	33	46	58
						386593	25	25	32	41	95
386593	12.5	26	33	43	74
						386593	6.25	28	31	43	53
386628	50	28	68	44	76
						386628	25	24	32	40	57
386628	12.5	28	35	42	75
						386628	6.25	22	29	40	59
386594	50	29	34	46	92
						386594	25	27	31	47	82
386594	12.5	28	33	45	74
						386594	6.25	23	48	42	67

Table 33: organ weight's (accounting for the percent of saline control)

ISIS NO	Dosage (mg/kg)	Liver	WAT	Kidney	Spleen
						Saline	N/A	100	100	100	100
148364	50	103	80	108	123
						148364	25	103	75	112	115
148364	12.5	100	84	108	96
						148364	6.25	101	89	104	113
386626	50	112	77	104	130
						386626	25	109	97	103	120
386626	12.5	96	73	97	114
						386626	6.25	100	90	100	95

ISIS NO	Dosage (mg/kg)	Liver	WAT	Kidney	Spleen
						386627	50	90	113	102	165
386627	25	99	87	99	143
						386627	12.5	109	93	102	136
386627	6.25	103	96	102	131
						386593	50	96	98	102	118
386593	25	83	94	100	104
						386593	12.5	99	82	101	129
386593	6.25	96	77	98	144
						386628	50	104	100	99	126
386628	25	102	97	109	113
						386628	12.5	101	111	99	114
386628	6.25	98	106	102	151
						386594	50	90	80	99	131
386594	25	93	76	99	128
						386594	12.5	94	98	100	113
386594	6.25	102	85	101	119

Generally speaking, the GCGR antisense compounds seldom shows or does not show fully adverse side effect.

Table 34: triglyceride (TRIG), cholesterol (CHOL) and glucose level (IU/L)

ISIS NO	Dosage (mg/kg)	TRIG Pre-Bleed	TRIG Post-Bleed	CHOL Pre-Bleed	CHOL Post-Bleed	Glucose Pre-Bleed	Glucose Post-Bleed
								Saline	N/A	132	181	91	96	208	285
148364	50	110	177	81	94	207	228
								148364	25	115	200	83	96	219	239
148364	12.5	106	179	85	89	198	256
								148364	6.25	86	162	86	89	226	215
386626	50	87	163	79	57	239	179
								386626	25	100	187	87	72	235	186
386626	12.5	100	148	82	76	232	185
								386626	6.25	86	162	85	90	222	221
386627	50	106	120	83	126	227	150
								386627	25	101	148	90	115	218	203
386627	12.5	99	203	86	98	237	219
								386627	6.25	111	165	88	104	238	228
386593	50	130	128	100	95	244	213
								386593	25	119	135	83	77	206	208
386593	12.5	122	128	83	79	222	233
								386593	6.25	120	138	84	78	214	219
386628	50	102	98	88	95	209	232
								386628	25	102	129	84	85	210	223
386628	12.5	90	123	90	94	231	240
								386628	6.25	117	121	83	85	228	229
386594	50	93	99	84	85	203	274

ISIS NO	Dosage (mg/kg)	TRIG Pre-Bleed	TRIG Post-Bleed	CHOL Pre-Bleed	CHOL Post-Bleed	Glucose Pre-Bleed	Glucose Post-Bleed
								386594	25	106	94	90	86	219	272
386594	12.5	118	133	85	95	200	292
								386594	6.25	112	146	78	94	222	275

GCGR 2-10-2MOE gapmer demonstrates the trend that trends towards blood-letting triglyceride levels after the low processing for 5-10-5MOE gapmer, and wherein ISIS 386628 and ISIS 386594 have maximum dose dependent effect.After handling with ISIS 386626 and ISIS386627, glucose level also descends in the dose dependent mode.Handle the decline that also mainly causes handling back blood-letting glucose level with ISIS386628, ISIS 386593 and ISIS 386594.As if cholesterol levels do not have significant difference throughout between the reason group.

For whether the phenotype variation shown in more than determining is relevant with the reduction of GCGR mRNA, assess the level of target mRNA in liver by method described herein to being subject to processing animal by PCR in real time.What table 35 and 41 showed is the direct comparative result of the antisense compounds of targeting GCGR nucleic acid to the effect of target expression.The result represents with the percent that accounts for saline control.

Table 35: with the GCGR mRNA level after ISIS 148364 and ISIS 386626 processing

ISIS NO	50mg/kg	25mg/kg	12.5mg/kg	6.25mg/kg
					148364	36	79	87	62
386626	0	8	3	7

Table 36: with the GCGR mRNA level after ISIS 148364 and ISIS 386627 processing

ISIS NO	50mg/kg	25mg/kg	12.5mg/kg	6.25mg/kg
					148364	63	87	105	86
386627	3	30	57	74

Table 37: with the GCGR mRNA level after ISIS 148364 and ISIS 386593 processing

ISIS NO	50mg/kg	25mg/kg	12.5mg/kg	6.25mg/kg
					148364	56	74	105	86
386593	9	38	74	90

Table 38: with the GCGR mRNA level after ISIS 148364 and ISIS 386628 processing

ISIS NO	50mg/kg	25mg/kg	12.5mg/kg	6.25mg/kg
					148364	42	77	98	101
386628	2	18	53	77

Table 39: with the GCGR mRNA level after ISIS 148364 and ISIS 386594 processing

ISIS NO	50mg/kg	25mg/kg	12.5mg/kg	6.25mg/kg
					148364	59	98	102	96
386594	25	47	50	96

Table 40: with the GCGR mRNA level after ISIS 386627 and ISIS 386593 processing

ISIS NO	50mg/kg	25mg/kg	12.5mg/kg	6.25mg/kg
					386627	5	40	58	42
386593	10	29	34	71

Table 41: with the GCGR mRNA level after ISIS 386628 and ISIS 386594 processing

ISIS NO	50mg/kg	25mg/kg	12.5mg/kg	6.25mg/kg
					386628	4	13	38	97
386594	19	50	56	99

Handling the remarkable dose dependent that has caused GCGR mRNA to express with 2-10-2MOE gapmer descends.ISIS 386626 demonstrates maximum target mRNA and descends.For determining whether improve in the observed efficient of short antisense compounds is because due to the increase of antisense compounds accumulation, measured the total antisense compounds concentration in liver and the kidney.

Table 42: the summation total length antisense compounds concentration (μ g/g) in liver and the kidney

ISIS NO	Total kidney	Total liver	The total length kidney	The total length liver
					148364	90	54	58	46
386626	757	274	355	125

ISIS NO	Total kidney	Total liver	The total length kidney	The total length liver
					386593	91	12	77	12
386628	496	286	305	202

Result shown in the table 42 proves, comprises C₁₆The short antisense compounds of conjugate is presented at the remarkable increase that all occurs the antisense compounds accumulation in liver and the kidney.But for the ISIS 386593 that can effectively reduce target mRNA, triglyceride and glucose level, the accumulating level in liver is similar to 5-10-5 MOE gapmer, and accumulating level is lower in kidney.Though these results suggest are and C₁₆Binding energy improve liver and kidney antisense compounds concentration, but this and not exclusively be the reason that causes the effectiveness of short antisense compounds.

These results prove that generally the short antisense compounds of GCGR can significantly suppress target mRNA expresses, and goes back triglyceride reducing and glucose level simultaneously.In addition, except ISIS386627, each short MOE gapmer seldom shows and even does not show toxic action.

Embodiment 5: targeting GCGR nucleic acid and have 2 '-MOE and methylene oxygen base (4 '-CH₂-O-2 ') the short antisense compounds of BNA modification

By intraperitoneal (i.p.) injection give eight ages in week male C57/BL6 mice (JacksonLaboratory, Bar Harbor, ME) the GCGR antisense compounds of single dose, concentration is 10,3.2,1 and 0.32 μ mol/kg.Each dosage group has four mices.Shown in the table 43 is sequence, die body and the conjugate that is used for the GCGR antisense compounds of this research.The runic residue represents that (2 '-MOE) modifies 2 '-O-methoxy ethyl, and the italic residue is represented methylene oxygen base (4 '-CH₂-O-2 ') BNA modifies.All antisense compounds all are to comprise between the thiophosphate nucleoside to connect key without exception, and each cytosine is replaced by 5-methylcytosine.The GCGR nucleic acid that GCGR antisense compounds energy targeting has been announced comprises Genbank searching number BC031885.1 (SEQID NO:7).

Table 43: the antisense compounds of targeting GCGR nucleic acid

ISIS NO	Target SEQ ID NO	5 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						148364	7	393	TGCACTTTGTGGTACCAAGG	5-10-5MOE	1562
396144	7	1768	GCTTCTCCATCATA	2-10-2MOE	1566
						396148	7	1768	GCTTCTCCATCATA	2-10-2 methylene oxygen base (4 '-CH2-O-2’) BNA	1567
396145	7	1765	ATGGCTTCTCCATCATATCC	5-10-5MOE	1568
						396146	7	1244	GGGCATGCTCGTCA	2-10-2MOE	650
396149	7	1244	GGGCATGCTCGTCA	2-10-2 methylene oxygen base (4 '-CH2-O-2’) BNA	652
						396147	7	1241	CTTGGGCATGCTCGTCAGTC	5-10-5MOE	1569

For whether the phenotype variation shown in more than determining is relevant with the reduction of GCGR mRNA, assess the level of target mRNA in liver by method described herein to being subject to processing animal by RT, PCR in real time.What table 44 showed is the direct comparative result of the antisense compounds of targeting GCGR nucleic acid to the effect of target expression.The result represents with the percent that accounts for saline control.

Table 44:GCGRmRNA level

ISIS NO.	0.32μmol/kg	1μmol/kg	3.2μmol/kg	10μmol/kg
					148364	105	106	73	38
396144	122	117	40	35
					396148	20	6	2	1
396145	nd	Nd	33	8
					396146	98	135	95	35
396149	91	41	30	7
					396147	nd	Nd	68	28

Shown in table 44, each has methylene oxygen base (4 '-CH₂-O-2 ') the short antisense compounds of BNA modification shows that all the dose dependent of GCGR mRNA level reduces.In addition, these short antisense compounds more can cause target to reduce than 5-10-5MOE gapmer effectively.Each comprises methylene oxygen base (4 '-CH in the pterion₂-O-2 ') the short antisense compounds of BNA all causes GCGR albumen for saline control and ISIS 148364 processing remarkable decline to be arranged.Then, calculate the estimation ED of each antisense compounds with Graphpad Prism₅₀Concentration; ED₅₀Be the dosage when observing 50%mRNA decline.The result shows in following table 45.

Table 45: the ED of estimation₅₀Concentration

The Gapmer die body	ISIS NO	ED50(μmole/kg)	ED50(mg/kg)
				5-10-5MOE	148364	7	50.6
2-10-2MOE	396144	4	18.1
				2-10-2 methylene oxygen base BNA	396148	0.1	0.4
5-10-5MOE	396145	2.1	9.3
				2-10-2MOE	396146	8.3	40
2-10-2 methylene oxygen base BNA	396149	1.1	5
				5-10-5MOE	396147	5.2	37.5

Embodiment 6: targeting PTEN nucleic acid and have methylene oxygen base (4 '-CH₂-O-2 ') the short antisense compounds of BNA modification

Give six ages in week male Balb/c mice (Jackson Laboratory, Bar Harbor ME) give single intraperitoneal injection PTEN antisense compounds, and dosage is 8 μ mol/kg.Each dosage group has four mices.Shown in the table 46 is sequence and the die body that is used for the PTEN antisense compounds of this research.The runic residue represents that (2 '-MOE), the italic residue is represented methylene oxygen base BNA nucleotide to 2 '-O-methoxy ethyl part.Each antisense compounds all is to comprise thiophosphate to connect key without exception.In addition, in the interval of ISIS 384073 and the cytosine residue in the pterion of ISIS 392056, ISIS 392057, ISIS 392061 and ISIS 392063 replaced by 5-methylcytosine.The PTEN nucleic acid that antisense compounds energy targeting has been announced comprises Genbank searching number U92437.1 (SEQ ID NO:13).

Table 46: the antisense compounds of targeting PTEN nucleic acid

ISIS NO	Target SEQ ID NO	5 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						141923	Contrast	N/A	CCTTCCCTGAAGGTTCCTCC	5-10-5MOE	1570
116847	29	2011	TCAAATCCAGAGGCTAGCAG	5-10-5MOE	1571
						384073	29	2013	AAATCCAGAGGCTAGC	3-10-3 methylene oxygen base (4 '-CH2-O-2’)BNA	1428
391172	29	2013	AAATCCAGAGGCTAG	2-10-3 methylene oxygen base (4 '-CH2-O-2’)BNA	1429

ISIS NO	Target SEQ ID NO	5 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						392056	29	140	AGCTGCAGCCATGA	2-10-2 methylene oxygen base (4 '-CH2-O-2’)BNA	1263
392057	29	807	GGTCCAGGGCCAAG	2-10-2 methylene oxygen base (4 '-CH2-O-2’)BNA	1162
						392061	29	2014	AATCCAGAGGCTAG	2-10-2 methylene oxygen base (4 '-CH2-O-2’)BNA	1431
392063	29	3099	AGGCCAGTGCTAAG	2-10-2 methylene oxygen base (4 '-CH2-O-2’)BNA	1226

Mice was put to death in injection in back 72 hours, with according to program as herein described and known to a person of ordinary skill in the art, measured serum transaminase level (table 47); Liver and spleen weight (table 47); And the PTEN mRNA level (table 48) in liver, kidney and the fat.

Table 47: transaminase level and organ weight

ISIS NO	AST (IU/L)	ALT (IU/L)	Liver weight (accounting for the percent of saline control)	Spleen weight (accounting for the percent of saline control)
					Saline	98.5	37.5	100	100
141923	89.5	34.8	101	108
					116847	59.8	29.5	109	108
384073	57.8	29.3	115	111
					391172	48.5	32.8	120	112
392056	516	892	125	167
					392057	63.8	34.5	125	101
392061	189	42.0	123	111
					392063	67.3	21.8	127	134

Generally speaking, have methylene oxygen base (4 '-CH₂-O-2 ') the short antisense compounds of BNA modification seldom shows and even does not show untoward reaction.In addition, TBW does not have significant difference between the reason group throughout.

Table 48: the %PTEN mRNA level in liver, kidney and the fat

ISIS NO	Liver	Kidney	Fat
				Saline	100	100	100
141923	102	133	118
				116847	37	96	85
384073	24	74	77
				391172	18	63	101
392056	27	88	74

ISIS NO	Liver	Kidney	Fat
				392057	33	79	96
392061	24	61	85
				392063	6.5	52	72

Shown in table 48, the target mRNA level that the antisense compounds of each targeting PTEN nucleic acid all causes comparing with the control treatment animal in the liver with saline treatment significantly reduces.Antisense compounds is various to the effect of the target mRNA level in kidney and the fat.

Embodiment 7: targeting PTEN nucleic acid and have the short antisense compounds that BNA modifies

Give six ages in week male Balb/c mice (Jackson Laboratory, Bar Harbor ME) give single intraperitoneal (i.p.) injection targeting PTEN the antisense compounds of nucleic acid, and dosage is 8,4,2 or 1 μ mol/kg.Each dosage group has four mices.Shown in the table 49 is sequence, pterion chemical constitution and the die body that is used for each antisense compounds of this research.The runic residue represents that tilted letter is represented methylene oxygen base (4 '-CH through the nucleotide of 2 '-MOE modification₂-O-2 ') BNA modifies.All antisense compounds all are to comprise thiophosphate in each position to connect key.Methylene oxygen base (4 '-CH of each cytosine of ISIS116847 and ISIS 392063₂-O-2 ') the cytosine residue in the BNA pterion is substituted by 5-methylcytosine, and methylene oxygen base (4 '-CH of ISIS 392745₂-O-2 ') thymidine in the BNA pterion is substituted by the 5-methylthymidine.The PTEN nucleic acid that antisense compounds energy targeting has been announced comprises Genbank searching number U92437.1 (SEQID NO:13).

Table 49: the antisense compounds of targeting PTEN nucleic acid

ISIS NO	Target SEQ ID NO	The target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						116847	13	2011	TCAAATCCAGAGGCTAGCAG	5-10-5 MOE	1571
392063	13	3099	CTTAGCACTGGCCT	2-10-2 methylene oxygen base BNA	1226
						392745	13	3099	CTTAGCACTGGCCT	The 2-10-2 methylene	1226

ISIS NO	Target SEQ ID NO	The target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
										Oxygen base BNA

Mice was put to death in injection in back 72 hours, to measure serum transaminase level (table 50); Liver, kidney and spleen weight (table 50); PTEN mRNA level (table 51) in the liver; And the ED that estimates₅₀Oligonucleotide concentration (table 52).These terminal points are to measure with method described herein and known to a person of ordinary skill in the art.

Table 50:AST, ALT and bilirubin level and organ weight

ISIS NO	Dosage μ mol/kg	AST (IU/L)	ALT (IU/L)	Bilirubin (mg/dL)	Liver weight (accounting for the percent of saline control)	Kidney weight (accounting for the percent of saline control)	Spleen weight (accounting for the percent of saline control)
								Saline	N/A	64.0	31.8	0.15	100	100	100
116847	8	73.0	32.0	0.1	114	92	106
								392063	8	50.3	17.3	0.1	115	98	115
392063	4	100.8	31.3	0.15	122	94	116
								392063	2	60.5	32.8	0.1	112	99	106
392063	1	57.5	29.3	0.1	104	95	107
								392745	8	75.5	23.5	0.13	125	99	100
392745	4	77.0	29.3	0.13	121	100	96
								392745	2	69.0	32.0	0.13	110	98	103
392745	1	52.0	27.3	0.1	109	97	104

Generally speaking, the PTEN antisense compounds does not demonstrate significant toxicity sign.Kidney, liver and spleen weight are all in normal range.TBW does not have significant difference between the reason group throughout.

Table 51: the %PTEN mRNA level (with respect to saline control) in the liver

ISIS NO	8μmol/kg	4 μmol/kg	2μmol/kg	1μmol/kg
					11684 7	36	ND	ND	ND
39206	7.4	16	32	60

ISIS NO	8 μmol/kg	4 μmol/kg	2 μmol/kg	1μmol/kg
					3
39274 5	5.2	11	31	60

Shown in table 51, each has methylene oxygen base (4 '-CH₂-O-2 ') the short antisense compounds of BNA modification shows that all the dose dependent of PTEN mRNA level reduces.In addition, these short antisense compounds more can cause target to reduce than 5-10-5MOE gapmer effectively.After the dosage with 8 μ mol/kg gives each antisense compounds, also measured the pten protein level in the liver.Each comprises methylene oxygen base (4 '-CH in the pterion₂-O-2 ') the short antisense compounds of BNA all causes pten protein for saline control and ISIS 116847 processing remarkable reduction to be arranged.Then, calculate the estimation ED of each oligonucleotide with Graphpad Prism₅₀Concentration.The result shows in following table 52.

Table 52: the ED of estimation₅₀Concentration

The pterion chemical constitution	ISIS NO	ED50(μmole/kg)	ED50(mg/kg)
				MOE (band 5-MeC)	116847	6.3	45.2
Methylene oxygen base BNA (band 5-MeC)	392063	1.3	5.8
				Methylene oxygen base BNA	392745	1.2	5.6

Be the further dissimilar dicyclic ring acid compound of research, designed the short antisense compounds of another group targeting PTEN nucleic acid and test.Give six ages in week male Balb/c mice (Jackson Laboratory, Bar Harbor ME) give single intraperitoneal (i.p.) injection antisense compounds, and dosage is 8,4,2 or 1 μ mol/kg.Each dosage group has four mices.Shown in the table 53 is sequence, pterion chemical constitution and the die body that is used for each antisense compounds of this research.All antisense compounds all are to comprise thiophosphate in each position to connect key.Methylene oxygen base (4 '-CH of ISIS392063₂-O-2 ') the cytosine residue in the BNA pterion is substituted by 5-methylcytosine.The PTEN nucleic acid that antisense compounds energy targeting has been announced comprises Genbank searching number U92437.1 (SEQ ID NO:13).

Table 53: the antisense compounds of targeting PTEN nucleic acid

ISIS NO	Target SEQ ID NO	5 ' target site	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
						392063	29	3099	CTTAGCACTGGCCT	2-10-2 methylene oxygen base BNA	1226
396564	29	3099	CTTAGCACTGGCCT	2-10-2 oxygen base amino (4 '-CH2-N(R)-O-2’) BNA	1226
						396006	29	3099	CTTAGCACTGGCCT	2-10-2 α-L-methylene oxygen base BNA	1226

Mice was put to death in injection in back 72 hours, with according to method as herein described and known to a person of ordinary skill in the art, measured serum transaminase level (table 54); Liver and spleen weight (table 54); And the PTEN mRNA level (table 55) in the liver.

Table 54:AST and ALT level and organ weight

ISIS NO	Dosage μ mol/g	AST (IU/L)	ALT (IU/L)	Liver weight	Spleen weight
						Saline	N/A	71	33	100	100
392063	8	97	38	118	103
						392063	4	179	36	115	107
392063	2	67	32	109	116
						392063	1	68	27	102	105
396564	8	67	25	100	104
						396564	4	96	30	102	106
396564	2	68	27	100	119
						396564	1	79	39	97	109
396006	8	56	28	110	104
						396006	2	139	36	97	105

Table 55: the %PTEN mRNA level (with respect to saline control) in the liver

ISIS NO	8μmol/kg	4μmol/kg	2μmol/kg	1μmol/kg
					392063	6.9	18	39	71
396564	86	97	100	96
					396006	6.5	ND	ND	70

As mentioned above, has α-L-methylene oxygen base (4 '-CH₂-O-2 ') the short antisense compounds of BNA modification has caused the dose dependent of target mRNA level to descend.The short antisense compounds that the amino BNA of apparatus aerobic base modifies is handled and has been caused the target expression to have appropriateness to descend.

Embodiment 8: single gives, the dose response research of carrying out with the short antisense compounds of targeting ApoB and PTEN nucleic acid

Give six ages in week male Balb/c mice (Jackson Laboratory, Bar Harbor ME) give single intraperitoneal (i.p.) injection antisense compounds, and dosage is 8,4,2 or 1 μ mol/kg.Each dosage group has four mices.Shown in the table 56 is sequence, pterion chemical constitution and the die body that is used for each antisense compounds of this research.The italic residue is represented methylene oxygen base (4 '-CH₂-O-2 ') BNA modifies, and the underscore residue is represented N-methyl-oxygen base amino (4 '-CH₂-N (CH₃)-O-2 ') BNA modifies, and adds the frame residue and represents α-L-methylene oxygen base (4 '-CH₂-O-2 ') BNA modifies.All antisense compounds all are to comprise thiophosphate in each position to connect key.Methylene oxygen base (4 '-CH of each cytosine of ISIS 116847 and ISIS 392063₂-O-2 ') the cytosine residue in the BNA pterion is substituted by 5-methylcytosine, and methylene oxygen base (4 '-CH of ISIS 392745₂-O-2 ') thymidine (thymidine) in the BNA pterion is substituted by 5-methylthymidine (thymidine).The PTEN nucleic acid that PTEN antisense compounds energy targeting has been announced comprises Genbank searching number U92437.1 (SEQ ID NO:13).The ApoB nucleic acid that ApoB antisense compounds energy targeting has been announced comprises Genbank searching number XM_137955.5 (SEQ ID NO:2).

Table 56: the short antisense compounds of targeting ApoB and PTEN nucleic acid

Table 57:ApoB and PTEN mRNA decline percent (with respect to saline control)

ISIS NO	Dosage (μ mol/kg)	ApoB mRNA decline percent (with respect to saline)	PTEN mRNA decline percent (with respect to saline)
				387462	8	0.62	92.8
	4	6.55	103
					2	18.6	105
	1	42.0	98.0
				392063	8	126	6.79
	4	111	18.1
					2	112	42.4
	1	114	62.3
				396565	8	116	23.8
	4	1.04	46.6
					2	94.4	76.1
	1	115	89.5
				396006	8	94.3	62.9
	4	101	18.2
					2	79.7	52.4
	1	111	82.4

Shown in table 57, each short antisense compounds with methylene oxygen base BNA modification shows that all the dose dependent of target mRNA level reduces.Particularly, the short antisense compounds (ISIS 396565) with the amino BNA of N-methyl-oxygen base pterion also demonstrates the dose dependent reduction of expressing with β-D-methylene oxygen base BNA and the similar PTEN of the short antisense compounds of α-L-methylene oxygen base BNA.Then, calculate the estimation ED of each antisense compounds with Graphpad Prism₅₀Concentration.The result shows in following table 58.

Table 58: the ED of estimation₅₀Concentration

The pterion chemical constitution	ISIS NO	ED50(μmole/kg)	ED50(mg/kg)
				Methylene oxygen base BNA	387462	0.8	3.9
Methylene oxygen base BNA	392063	1.5	7
				The amino BNA of N-Me-oxygen base	396565	3.8	17.4

The pterion chemical constitution	ISIS NO	ED50(μmole/kg)	ED50(mg/kg)
				α-L-methylene oxygen base BNA	396006	2.1	9.3

Embodiment 9: the parent antisense compounds of targeting SGLT-2mRNA and giving of parent mixed matrix antisense compounds

With ISIS 257016 with 1,7.5,14 or the dosage of 17mg/kg weekly twice intraperitoneal give the db/db mice (Charles River Laboratories, Wilmington, MA).Matched group comprises by identical dosage accepts brinish group and accept the group of ISIS 145733.ISIS257016 and ISIS 145733 comprise sequence GAAGTAGCCACCAACTGTGC (SEQ ID NO:1572), further comprise central authorities' " at interval " zone that 10 2 '-deoxyribonucleotides are formed, both sides (5 ' and 3 ' direction) at interval are connected to " pterion " of 5 nucleic acid.(2 '-MOE) nucleotide is formed by 2 '-methoxy ethyl in the pterion.All cytidine residues all are the 5-methylcytidines.For ISIS 145733, (skeleton) Lian Jian is thiophosphate (P=S) without exception between nucleoside; And ISIS 257016 has mixed matrix.Connecting key between the nucleoside of ISIS 257016 is di-phosphate ester (P=O) in the pterion, is thiophosphate in the interval.After giving last dosage 48 hours, mice is put to death, analyze the SGLT-2mRNA level of renal tissue.The result shows in following table 59.

The Antisense Suppression that table 59:5-10-5MOE gapmer expresses SGLT2mRNA in the body

Than saline control, ISIS 257016 and ISIS 145733 have significantly reduced the SGLT-2 level.(the mRNA level is to measure with RT, PCR in real time as mentioned above).But, proved that ISIS 257016 is reducing aspect the SGLT-2mRNA than ISIS 145733 powerful about 20-50 times.The reason group is also seen the relevant decline (the saline group is 661 ± 14, and the group of accepting ISIS 257016 is 470 ± 23) of plasma glucose levels throughout.ISIS 257016 and ISIS145733 being accumulated in the whole dosage range in kidney is all similar, but detects few total length 257016 antisense compounds in kidney, and this has supported that catabolite is this theory that causes the active reason that improves.And the onset that single gives behind the 25mg/kg is relevant with the time point (time pint) that does not almost have complete (intact) 257016 antisense compounds to stay.

By and the similar scheme of above-mentioned dosage, very lean mice, ob/ob mice with in ZDF rat (Charles Rivers Laboratories), carried out similar research.The sequence of the binding site of ISIS145733 and ISIS 257016 is (referring to table 60) of guarding between mice and rat.SGLT-2mRNA in the liver descends with above being seen similar.In the research of carrying out with rat, giving for twice weekly in continuous two weeks under the dosage of 10mg/kg, ISIS 145733 demonstrates and the SGLT-2mRNA level is reduced about 40%, and the reduction that ISIS257016 realized is greater than 80%.ISIS 257016 reduces the degree maximum that SGLT2 expresses under the low dosage of 12.5mg/kg.That carries out under than the low dosage scope studies show that in addition the mixed matrix antisense compounds causes the SGLT2mRNA level significantly to reduce under the dosage less than 1mg/kg/ week.

Embodiment 10: the parent antisense compounds of targeting SGLT-2mRNA gives with short antisense compounds

Pharmacokinetic shows that ISIS 257016 works as prodrug, and it becomes the pharmacophore of 12 nuclear bases through metabolism.In next one research, give ISIS 257016 or the ISIS 370717 that gives 1.5mg/kg in the ZDF rat peritoneum twice weekly, perhaps give saline with similar dosage.ISIS 370717 is antisense compounds of 12 nuclear bases of targeting SGLT-2 nucleic acid, comprise sequence TAGCCACCAACT (SEQ ID NO:154), and further comprising central authorities' " at interval " zone that 10 2 '-deoxyribonucleotides are formed, both sides (5 ' and 3 ' direction) at interval are connected to " pterion " of 1 nucleic acid.(2 '-MOE) nucleotide is formed by 2 '-methoxy ethyl in the pterion.All cytidine residues all are the 5-methylcytidines.(skeleton) Lian Jian is thiophosphate (P=S) without exception between the nucleoside of whole oligonucleotide.

5 week of administration back execution animal, analyze the SGLT-2mRNA level of renal tissue.ISIS 257016 is similar with the pharmacological activity of ISIS 370717, but the antisense compounds of these 12 nucleotide demonstrates onset faster.ISIS 370717 after single gives 2.8 μ mol/kg second day demonstrates SGLT2 in the kidney and expresses and almost have 80% to suppress, and ISIS257016 gives only to demonstrate about 25% in back second day at identical single and suppresses.Data acknowledgement ISIS 257016 is the prodrugs with 12 nucleotide pharmacophore.

Embodiment 11: the usefulness and the bioavailability of short antisense compounds

The improvement of the oral administration biaavailability of the performance improvements that ISIS 370717 is shown and these short antisense compounds makes them can be used for orally give.By allowing normal rat accept ISIS 370717, ISIS 145733 or the saline of 100mg/kg weekly for twice in the jejunum.Give back 48 hours execution animals for the last time, analyze the antisense compounds concentration and the SGLT-2mRNA level of renal tissue.Compared with the control, the accumulation of ISIS 370717 in renal tissue significantly higher (about 500 milligrams/gram tissue).In addition, SGLT-2mRNA has reduced above 80% than contrast.

Embodiment 12:12 the short antisense compounds of nucleotide pterion, interval and total length on every side changes

The relevant oligonucleotide of sequence that has the die body of variation with ISIS 3707171-10-1MOE gapmer as template for preparing.These variations provide in table 60.Use the sequence of having announced (to be respectively GenBank searching number U29881.1, to be incorporated herein with SEQ ID NO:1575; With GenBank searching number AJ292928.1, be incorporated herein with SEQ ID NO:1576), antisense compounds is designed to can the targeting mice or the zones of different of rat SGLT2 nucleic acid.

Table 60: the short antisense compounds of targeting SGLT2 nucleic acid

ISIS NO	5 ' target site on the mice SEQ ID NO:1575	5 ' target site on the rat SEQ ID NO:1576	The Gapmer die body	Sequence (5 '-3 ')	SEQ ID NO
						257016	2680	148	5-10-5 MOE	GAAGTAGCCACCAACTGTGC	1553
370717	2684	152	1-10-1 MOE	TAGCCACCAACT	1554
						386169	2684	152	2-8-2 MOE	TAGCCACCAACT	1555
386176	2685	153	1-8-1 MOE	AGCCACCAAC	1556
						386196	2684	152	3-6-3 MOE	TAGCCACCAACT	1557

Analyze the effect of antisense compounds to mice SGLT2mRNA level.Data are to take from the numerical range of three experiments (range), give by peritoneal injection to give 2.5,0.5 or the above-mentioned MOE gapmer of 0.1umol/kg continuous three weeks of mice weekly for twice in experiment.Give back 48 hours execution mices for the last time, the SGLT2 level in the assessment kidney.The SGLT2mRNA level is measured by RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.The result shows in following table 61.

Table 61:1-10-1 and 1-10-2MOE gapmer are to the Antisense Suppression of SGLT2 in the body

All various die bodys that tried of these presentation of results suppress in the dose dependent mode all that SGLT2 expresses in the body.Find that 1-10-1,2-8-2 and 1-8-1gapmer are potent especially.

Embodiment 13:1-10-1 and 1-10-2MOE Gapmer are to the Antisense Suppression of rat SGLT-2

1-10-1 that provides in the table 62 and 1-10-2MOE gapmer antisense compounds are designed to the zones of different of targeting mice or rat SGLT2 RNA.The short antisense compounds of in the table 62 all is the chimeric oligonucleotide (gamper) of length 12 or 13 nucleotide, it comprises central authorities' " at interval " section that 10 2 '-deoxyribonucleotides are formed, 5 ' side joint at interval has " pterion " of 1 nucleoside, and 3 ' side joint has " pterion " of 2 or 1 nucleotide.(2 '-MOE) nucleotide is formed by 2 '-methoxy ethyl in the pterion.(skeleton) Lian Jian is thiophosphate (P=S) between the nucleoside in the oligonucleotide.All cytidine residues all are the 5-methylcytidines.

Table 62: the antisense compounds of targeting SGLT2 nucleic acid

ISIS NO	5 ' target site (mice) on the SEQ ID NO:XXX	5 ' target site (rat) on the SEQ ID NO:XXX	The Gapmer die body	Sequence (5 '-3 ')	SEQ ID NO
						370717	2684	152	1-10-1 MOE	TAGCCACCAACT	1554
382675	2683	151	1-10-1 MOE	TAGCCACCAACTG	1559
						379692		508	1-10-1 MOE	TGTTCCAGCCCA	246
382676		507	1-10-2 MOE	TGTTCCAGCCCAG	246
						379699		1112	1-10-2 MOE	GGCATGAGCTT	281
382677		1111	1-10-2 MOE	GGCATGAGCTTCA	281
						382677		958	1-10-2 MOE	GGCATGAGCTTCA	281

Analyze of the effect of short antisense compounds to rat SGLT2mRNA level.Data are to take from the numerical range of three experiments, give by peritoneal injection to give 450,150 or 1-10-1 or the 1-10-2MOE gapmer of 50nmol/kg continuous three weeks of male Sprague-Dawley rat (170-200g) weekly for twice in experiment.Give back 48 hours execution rats for the last time, the SGLT2mRNA level in the assessment kidney.The target level is measured by RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.The result shows in following table 63.

Table 63:1-10-1 and 1-10-2MOE gapmer are to the Antisense Suppression of SGLT2mRNA in the body

These presentation of results 1-10-1 and 1-10-2MOE gapmer reduce SGLT2mRNA in the body in the dose dependent mode.

TBW and liver, spleen and the kidney weight of rat have also further been assessed.The significant change of spleen, liver weight or body weight can show that certain specific compound has caused toxic action.All variations are all in the range of error of experiment.In processing procedure or when research stops, do not observe significant body weight change.Not observing significant liver or spleen weight changes.

The toxic action of the short antisense compounds that gives in the body also can be assessed by measurement enzyme and the proteinic level relevant with the i or I of liver or kidney.The assessed value of the level of serum transaminase aspartate aminotransferase (AST) and alanine aminotransferase (ALT), the index of hepatic disease or damage often.Serum total bilirubin is the index of hepato-biliary function, and albumin and blood urea nitrogen (BUN) are the indexs of renal function.Glucose and triglyceride levels change because of the toxicity of handling sometimes.Serum glucose also depends on the activity of SGLT2 on the part.In the rat of handling, measure the level of ALT, AST, total bilirubin, albumin, BUN, glucose and triglyceride with short antisense compounds.The level of the routine clinical index of liver and kidney injury and disease does not have significant change with respect to the saline treatment animal in normal range, the short antisense compounds of this proof can appreciable impact kidney or liver function.Triglyceride and glucose level significantly do not raise for the saline treatment animal.

Embodiment 14:1-10-1MOE Gapmer is to the Antisense Suppression of mice and rat SGLT-2

Table 64 has shown the 1-10-1MOE gapmer antisense compounds of the zones of different that is designed to targeting mice SGLT2mRNA.

Table 64: the structure of the antisense compounds of targeting SGLT2mRNA is formed

ISIS NO	5 ' target site (mice) on the SEQ ID NO:XXX	5 ' target site (rat) on the SEQ ID NO:XXX	Die body	Sequence (5 '-3 ')	SEQ ID NO
						370717	2684	152	1-10-1MOE	TAGCCACCAACT	1554
379692		508	1-10-1MOE	TGTTCCAGCCCA	246
						379699		1112	1-10-1MOE	GGCATGAGCTTC	281
379702		1525	1-10-1MOE	GCACACAGCTGC	293
						381408	3034**		1-10-1MOE	TACCGAACACCT	1560

* represents that target sequence is had 3 mispairing

Analyze of the effect of short antisense compounds to mice SGLT2mRNA level.Data are taken from three experiments, give by peritoneal injection to give 450,150 or one of the above-mentioned 1-10-1MOE gapmer of 50nmol/kg male 6 weeks continuous two weeks of Balb/c mice in age weekly for twice in experiment.Give back 48 hours execution mices for the last time, the SGLT2mRNA level in the assessment kidney.The target level is measured by RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.The result shows in following table 65.

Table 65:1-10-1MOE gapmer is to the Antisense Suppression of SGLT-2mRNA in the body

These presentation of results, except ISIS 381408, all 1-10-1MOEgapmer suppress SGLT2 expression in the body in the dose dependent mode.The activity of ISIS381408 has shown (referring to table 65) in rat studies.

The assessment of 1-10-1 Gapmer in the rat

Studied the effect of above-mentioned 1-10-1gapmer (ginseng sees the above table 64) to rat SGLT2mRNA level.Data are taken from four experiments, continuous three weeks of male Sprague-Dawley rat (170-200g) are given 250nmol/kg twice weekly by peritoneal injection in experiment.Give back 48 hours execution rats for the last time, the SGLT2mRNA level in the assessment kidney.The target level is measured by RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.The result shows in following table 66.

Table 66:1-10-1MOE gapmer is to the Antisense Suppression of SGLT2mRNA in the body

These presentation of results, all 1-10-1MOE gapmer all suppress the SGLT2 expression in the rat studies in vivo.

Embodiment 15: other 1-10-1 and 2-8-2MOE Gapmer are to the Antisense Suppression of mice and rat SGLT-2 expression

1-10-1 and 2-8-2MOE gapmer are short, and antisense compounds is designed to the zones of different of targeting mice SGLT2RNA, but has complementarity between each species.Table 67 has shown each short antisense compounds.The short antisense compounds of all of table 67 all is the gapmer of 12 nucleotide of length, comprises central authorities' " at interval " section of being made up of 2 '-deoxyribonucleotide, and both sides (5 ' and 3 ' direction) at interval are connected to " pterion " modified with 2 '.(2 '-MOE) nucleotide is formed by 2 '-methoxy ethyl in the pterion.(skeleton) Lian Jian is thiophosphate (P=S) between the nucleoside in the oligonucleotide.All cytidine residues all are the 5-methylcytidines.

Table 67: the short antisense compounds of targeting SGLT2 nucleic acid

ISIS NO	5 ' target site (rat)	Target SEQ ID (rat)	The Gapmer die body	Sequence (5 '-3 ')	SEQ ID NO
						379692	508		1-10-1MOE	TGTTCCAGCCCA	246
388625	508		1-10-1MOE	TGTTCCAGCCCA	246
						379699	1112		1-10-1MOE	GGCATGAGCTTC	281
388626	1112		2-8-2MOE	GGCATGAGCTTC	281
						379702	1525		2-8-2MOE	GCACACAGCTGC	293
388627	1525		2-8-2MOE	GCACACAGCTGC	293

Analyze of the effect of short antisense compounds to SGLT2mRNA level in the mice body.Data are taken from three experiments, give by peritoneal injection to give 0.5,0.1 or 1-10-1 or the 2-8-2MOEgapmer of 0.02umol/kg male 6 weeks continuous three weeks of Balb/c mice in age weekly for twice in experiment.Give back 48 hours execution mices for the last time, the SGLT2 level in the assessment kidney.The target level is measured by RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.The result shows in following table 68.

Table 68:1-10-1 and 2-8-2MOE gapmer are to the Antisense Suppression of GLT2mRNA in the body

These presentation of results 1-10-1 and 2-8-2MOE gapmer are the expression that suppresses SGLT2 in the body in the dose dependent mode.

TBW and liver, spleen and the kidney weight of mice have also further been assessed.All variations are all in the range of error of experiment.In processing procedure or when research stops, do not observe significant body weight change.Not observing significant liver or spleen weight changes.

In the mice of handling, measured the level of ALT, AST, BUN, transaminase, plasma creatinine, glucose and triglyceride with short antisense compounds.The level of the routine clinical index of liver and kidney injury and disease does not have significant change with respect to the saline treatment animal in normal range, the short antisense compounds of this proof can appreciable impact kidney or liver function.Triglyceride and glucose level does not significantly raise for the saline treatment animal.

Assessment ISIS 3796921-10-1 MOE Gapmer, ISIS 3921701-10-1 methylene oxygen base BNA Gapmer, ISIS 3886252-8-2MOE Gapmer and ISIS 392173 2-8-2 methylene oxygen base BNA Gapmer in mice

ISIS 379692 1-10-1 MOE gapmer and ISIS 3886252-8-2MOEgapmer are compared (referring to table 69) to the effect of SGLT2mRNA level in the mice body and this effect of ISIS 392170 1-10-1 methylene oxygen base BNA Gapmer and ISIS 3921732-8-2 methylene oxygen base BNAGapmer.Data are taken from three experiments, give by peritoneal injection to give 5,25 and ISIS 3796921-10-1MOE gapmer or the ISIS 3886252-8-2MOE gapmer of 125nmol/kg male 6 weeks continuous three weeks of Balb/c mice in age weekly for twice in experiment.Give back 48 hours execution mices for the last time, the SGLT2mRNA level in the assessment kidney.The target level is measured by RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.Data are represented with the percent change ("+" expression increases, and "-" expression descends) with respect to the saline treatment animal, and are shown in table 69.

Table 69:1-10-1 and 2-8-2MOE gapmer are to the Antisense Suppression of SGLT2mRNA in the body

Oligonucleotide dosage nmol/kg	ISIS 379692 1-10-1MOE	ISIS 392170 1-10-1 methylene oxygen base BNA	ISIS 388625 2-8-2MOE	ISIS 392173 2-8-2 methylene oxygen base BNA
					125	-58	-69	-70	-75
25	-46	-54	-47	-57
					5	-7	-23	-18	-44

These presentation of results 1-10-1 and 2-8-2 MOE gapmer are the expression that suppresses SGLT2 in the body in the dose dependent mode in three the highest dosage ranges.The result illustrates that also methylene oxygen base BNA construction is more more potent than MOE construction.In processing procedure or when research stops, do not observe significant body weight change.Not observing significant liver or spleen weight changes.The toxicity parameter of level that comprises ALT, AST, BUN and kreatinin does not have significant change with respect to the saline treatment animal in normal range, this illustrates that these chemical compounds can appreciable impact kidney or liver function.

Assessment ISIS 379692 1-10-1 MOE Gapmer and ISIS 3886252-8-2 MOE Gapmer in rat

Assessed ISIS 3796921-10-1 MOE gapmer and ISIS 388625 MOE2-8-2 gapmer (referring to table 70) effect to SGLT2mRNA level in the rat body.Data are taken from four experiments, give by peritoneal injection to give 200,50,12.5 or ISIS 3796921-10-1 MOE gapmer or the ISIS 388625 2-8-2 MOEgapmer of 3.125nmol/kg continuous three weeks of male Sprague-Dawley rat (170-200g) weekly for twice in experiment.Give back 48 hours execution rats for the last time, the SGLT2 level in the assessment kidney.The target level is measured by RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.The result shows in following table 70.

Table 70:1-10-1 and 2-8-2 MOE gapmer are to the Antisense Suppression of SGLT2mRNA in the body

These presentation of results 1-10-1 and 2-8-2MOE gapmer are the expression that suppresses SGLT2 in the body in the dose dependent mode in three the highest dosage ranges.

TBW and liver, spleen and the kidney weight of rat have also further been assessed.All variations are all in the range of error of experiment.In processing procedure or when research stops, do not observe significant body weight change.Not observing significant liver or spleen weight changes.

In the rat of handling, measured the level of ALT, AST, BUN, cholesterol, plasma creatinine and triglyceride with short antisense compounds.The level of the routine clinical index of liver and kidney injury and disease does not have significant change with respect to the saline treatment animal in normal range, the short antisense compounds of this proof can appreciable impact kidney or liver function.

The Antisense Suppression that SGLT2 expresses in the embodiment 16:ZDF rat

Analyzed the effect of ISIS 388625,388626 and the gentle HbA1c of control oligonucleotide ISIS 388628 pairs of ZDF rat plasmas G/W.Leptin receptor defects type Zucker diabetes fat (ZDF) rat is the useful model of research type ii diabetes.Diabetes spontaneous generation when age in these male rats 8-10 week, and follow and hyperphagia, polyuria, excessive thirst and weightening finish occur and cut down, these symptoms similar to the clinical symptoms of diabetes (Phillips MS, et al., 1996, Nat Genet 13,18-19).Give six ZDF rat weekly peritoneal injections of continuous 12 weeks in age in week short antisense compounds, dosage is 400nM/kg.Data show in table 71 and 72.

Table 71: plasma glucose

Table 72:HbA1c state

Compare with the animal of PBS and control treatment, ISIS 388625 and 388626 has significantly reduced plasma glucose levels and HbA1C.

Embodiment 17: Antisense Suppression (ISIS 388625) ISIS 388625 that the dirty middle SGLT2 of Testis et Pentis Canis is expressed is 2-8-2MOE Gapmer, and its sequence isTGGTTCCAGCCCA(SEQ ID NO:246) (for example referring to table 71).Studied the effect of 388625 couples of Canis familiaris L. SGLT2mRNA of ISIS level.Data are taken from two dosage groups, have in two groups 9 male than lattice Canis familiaris L. (beagle dog), twice subcutaneous injection 1 or ISIS in 10mg/kg/ week 388625 or saline weekly.Put to death all Canis familiaris L.s at the 46th day that studies, assessment kidney SGLT2 level.The target level is measured by quantitative RT, PCR in real time as described in other embodiment of this paper.PCR result normalizes to inner ISIS contrast.The result shows in following table 73.

The Antisense Suppression of SGLT2mRNA in 388625 pairs of bodies of table 73:ISIS

These presentation of results can realize that SGLT2mRNA descends greater than 80% under the ISIS in 1mg/kg/ week 388625 dosage.Under high slightly dosage, can realize bigger decline.Also confirm in Canis familiaris L., to give ISIS 388625 and can improve glucose tolerance.In the glucose tolerance test of standard, to compare with saline control, the peak value plasma glucose levels on average reduces above 50%, and glucose subsequently descends and reduces.The urine glucose is drained and has also been increased.

Embodiment 18: the in vivo test of the short antisense compounds of targeting SGLT2 nucleic acid

Design, synthesized 20 with people/monkey/mice/complementary 1-10-1MOEgapmer of rat SGLT2, and in vivo test their suppress the situation of SGLT2mRNA level in kidney.Table 74 provides the target site to mice and rat.Table 4 and 5 provides the target site to the people.Data are meansigma methodss of two experiments, in experiment, give male 6 continuous two weeks of Balb/c mice in age in week the oligonucleotide of twice peritoneal injection (totally four injections) 350nmol/kg weekly.Give back 48 hours execution mices for the last time, the SGLT2mRNA level in the assessment kidney.The SGLT2mRNA level be with two different PCR primer probe groups---primer probe groups (PPS) 534 and PPS 553---according to the program of standard, measure by quantitative PCR in real time analysis.The SGLT2mRNA level is normalized to cyclophilin mRNA level, and back one level also is to measure by quantitative PCR in real time.The result shows in following table 74.

Table 74: the Antisense Suppression of SGLT2 in the body

ISIS

SEQ ID

SEQ ID

Sequence (5 '-3 ')

Die body

PPS

PPS

SEQ

NO	5 ' target site (mice) on the NO:XXX	5 ' target site (rat) on the NO:XXX			534 (accounting for the percent of saline control)	553 (accounting for the percent of saline control)	ID NO
								PBS			N/A		---	---
370717	2684	152	TAGCCACCAACT	1-10-1 MOE	-84.4	-84.3	1554
								379684	2070	64	TGTCAGCAGGAT	1-10-1 MOE	-45.0	-43.2	214
379685	2103	97	TGACCAGCAGGA	1-10-1 MOE	-10.3	-20.5	219
								379686	2121*	115	ACCACAAGCCAA	1-10-1 MOE	-71.9	-75.1	225
379687	2824	216	GATGTTGCTGGC	1-10-1 MOE	-47.1	-52.1	230
								379688	2876	268	CCAAGCCACTTG	1-10-1 MOE	-62.6	-70.4	240
379689		298	AGAGCGCATTCC	1-10-1 MOE	-17.5	-30.4	241
								379690		415	ACAGGTAGAGGC	1-10-1 MOE	-18.9	-22.5	242
379691		454	AGATCTTGGTGA	1-10-1 MOE	-35.0	-48.6	243
								379692		508	TGTTCCAGCCCA	1-10-1 MOE	-88.1	-88.5	246
379693		546	CATGGTGATGCC	1-10-1 MOE	-51.6	-59.9	254
								379694		609	GACGAAGGTCTG	1-10-1 MOE	-42.1	-54.4	264
379695		717	GGACACCGTCAG	1-10-1 MOE	-52.5	-64.1	266
								379696		954	CAGCTTCAGGTA	1-10-1 MOE	-24.6	-36.2	267
379697		982	CTGGCATGACCA	1-10-1 MOE	-32.0	-46.3	272
								379698		1071	GCAGCCCACCTC	1-10-1 MOE	-11.8	-27.0	275
379699		1112	GGCATGAGCTTC	1-10-1 MOE	-83.5	-85.8	281
								379700		1138	CCAGCATGAGTC	1-10-1 MOE	-2.8	-16.4	285
379701		1210	CCATGGTGAAGA	1-10-1 MOE	-0.3	-11.9	288
								379702		1525	GCACACAGCTGC	1-10-1 MOE	-87.8	-89.5	293
379703		1681	GCCGGAGACTGA	1-10-1 MOE	-44.2	-45.9	295

* representing has 1 or 2 mispairing to target sequence

The Antisense Suppression of people PCSK9 in the embodiment 19:Hep3B cell

Tested of the effect of the short antisense compounds of targeting PCSK9 nucleic acid to external PCSK9mRNA.Table 6 has shown each short antisense compounds.Isis No, Gapmer die body and the SEQ ID NO of each short antisense compounds show again in table 75.Short antisense compounds with 100mM is handled the Hep3B cell of cultivating.The 5-10-5MOEgapmer of targeting PCSK9 nucleic acid is as positive control.After the processing time section,, as described herein by quantitative PCR in real time measurement PCSK9mRNA level from cell separation RNA.With the PCSK9mRNA level according to passing through

Total rna content of measuring is adjusted.The result suppresses percent (%Inhib) expression with the PCSK9 with respect to untreatment control cell in table 75.In " (%Inhib) " hurdle, " 0 " represents that this specific antisense compounds do not observe the decline of PCSK9mRNA.

Table 75: short antisense compounds is to the Antisense Suppression of PCSK9

ISIS No.	SEQ ID NO	5 ' target site on the SEQ ID NO:4	3 ' target site on the SEQ ID NO:4	The Gapmer die body	% suppresses scope	％Inhib
							400297	329	695	708	2-10-2MOE		0
400298	330	696	709	2-10-2MOE		0
							400299	331	697	710	2-10-2MOE		0
400300	332	742	755	2-10-2MOE		9
							400301	333	757	770	2-10-2MOE	20-30％	27
400302	334	828	841	2-10-2MOE		0
							400303	335	829	842	2-10-2MOE		0
400304	336	830	843	2-10-2MOE	10-20％	11
							400305	337	937	950	2-10-2MOE	30-40％	38
400306	338	952	965	2-10-2MOE	40-50％	40
							400307	339	988	1001	2-10-2MOE	70-80％	76
400308	340	989	1002	2-10-2MOE	50-60％	55
							400309	341	990	1003	2-10-2MOE	40-50％	44
400310	342	991	1004	2-10-2MOE		8
							400311	343	992	1005	2-10-2MOE	10-20％	18
400312	344	993	1006	2-10-2MOE	20-30％	28
							400313	345	994	1007	2-10-2MOE	10-20％	10
400314	346	1057	1070	2-10-2MOE	20-30％	26
							400315	347	1075	1088	2-10-2MOE		0
400316	348	1076	1089	2-10-2MOE		8

400317	349	1077	1090	2-10-2MOE		7
							400318	350	1078	1091	2-10-2MOE	20-30％	26
400319	351	1093	1106	2-10-2MOE		0
							400320	352	1094	1107	2-10-2MOE		0
400321	353	1095	1108	2-10-2MOE		0
							400322	354	1096	1109	2-10-2MOE		0
400323	355	1147	1160	2-10-2MOE		0
							400324	356	1255	1268	2-10-2MOE		7
400325	357	1334	1347	2-10-2MOE		4
							400326	358	1335	1348	2-10-2MOE		0
400327	359	1336	1349	2-10-2MOE	30-40％	36
							400328	360	1453	1466	2-10-2MOE	10-20％	13
400329	361	1454	1467	2-10-2MOE	10-20％	14
							400330	362	1455	1468	2-10-2MOE	40-50％	43
400331	363	1456	1469	2-10-2MOE	30-40％	35
							400332	364	1569	1582	2-10-2MOE		0
400333	365	1570	1583	2-10-2MOE		0
							400334	366	1571	1584	2-10-2MOE		0
400335	367	1572	1585	2-10-2MOE		0
							400336	368	1573	1586	2-10-2MOE		4
400337	369	1574	1587	2-10-2MOE		0
							400338	370	1575	1588	2-10-2MOE		9
400339	371	1576	1589	2-10-2MOE		0
							400340	372	1577	1590	2-10-2MOE		0
400341	373	1578	1591	2-10-2MOE		0
							400342	374	1621	1634	2-10-2MOE		0
400343	375	1622	1635	2-10-2MOE		0
							400344	376	1623	1636	2-10-2MOE		0
400345	377	1624	1637	2-10-2MOE		0
							400346	378	1738	1751	2-10-2MOE		5
400347	379	1739	1752	2-10-2MOE		0
							400348	380	1740	1753	2-10-2MOE		0
400349	381	1741	1754	2-10-2MOE	10-20％	13
							400350	382	1834	1847	2-10-2MOE	10-20％	15
400351	383	1835	1848	2-10-2MOE	10-20％	14
							400352	384	1836	1849	2-10-2MOE	20-30％	29
400353	385	1837	1850	2-10-2MOE	10-20％	19
							400354	386	1838	1851	2-10-2MOE	10-20％	19
400355	387	1839	1852	2-10-2MOE		0
							400356	388	1840	1853	2-10-2MOE		0
400357	389	2083	2096	2-10-2MOE		0
							400358	390	2084	2097	2-10-2MOE	10-20％	12

400359	391	2085	2098	2-10-2MOE		0
							400360	392	2086	2099	2-10-2MOE	30-40％	38
400361	393	2316	2329	2-10-2MOE		2
							400362	394	2317	2330	2-10-2MOE	10-20％	16
400363	395	2318	2331	2-10-2MOE		8
							400364	396	2319	2332	2-10-2MOE		0
400365	397	2320	2333	2-10-2MOE	20-30％	25
							400366	398	2321	2334	2-10-2MOE	10-20％	15
400367	399	2322	2335	2-10-2MOE	10-20％	12
							400368	400	2323	2336	2-10-2MOE	10-20％	11
400369	401	2324	2337	2-10-2MOE		0
							400370	402	2325	2338	2-10-2MOE	10-20％	13
400371	403	3543	3556	2-10-2MOE		0

Shown in table 75, short antisense compounds targeting PCSK9 nucleic acid, that have 2-10-2MOE Gapmer die body has reduced the PCSK9 mRNA in the cultured cell.

The short antisense compounds of targeting PCSK9 nucleic acid is to test in Hep3B cell dosage response experiment.Cell is that the short antisense compounds of the nM concentration that provides with table 76 as described herein is handled.After the processing time section,, as described herein by quantitative PCR in real time measurement PCSK9mRNA level from cell separation RNA.The PCSK9mRNA level is normalized to cyclophilin mRNA level, and back one level is to measure by PCR in real time with cyclophilin Auele Specific Primer probe groups.The result suppresses percent with the PCSK9 with respect to untreatment control cell and represents.The EC that has also shown every kind of short antisense compounds of test in the dose response experiment₅₀(observe the mRNA reduction and reach 50% o'clock concentration), EC₅₀Be to calculate with Graphpad Prism.As shown in the table, the PCSK9mRNA level reduces in the dose dependent mode.

Table 76: short antisense compounds is to the dose dependent Antisense Suppression of PCSK9

Embodiment 20: comprise the Antisense Suppression of the short antisense compounds of BNAs to PCSK9

The short antisense compounds of targeting PCSK9 nucleic acid is to test in mice and people's cultured cell in the dose response experiment.The chemical compound that is tried comprises ISIS 403739 and ISIS403740.ISIS 403739 these short antisense compounds are made up of the nucleotide sequence of SEQ ID NO:404, have the 2-10-2Gapmer die body, and wherein the nucleotide in the pterion comprises (6 ' S)-6 ' methyl BNA.ISIS 403740 these short antisense compounds are made up of the nucleotide sequence of SEQ ID NO:405, have the 2-10-2Gapmer die body, and wherein the nucleotide in the pterion comprises (6 ' S)-6 ' methyl BNA.Also tested the 5-10-5MOE gapmer of targeting PCSK9 nucleic acid.

Mouse liver cell is inoculated, and the short antisense compounds of the nM concentration that provides with table 77 as described herein is handled.After the processing time,, as described herein by quantitative PCR in real time measurement PCSK9mRNA level from cell separation RNA.The PCSK9mRNA level is normalized to cyclophilin mRNA level, and back one level is to measure by PCR in real time with cyclophilin Auele Specific Primer probe groups.The result suppresses percent with the PCSK9 with respect to untreatment control cell and represents.Wherein the decline of PCSK9mRNA is not observed in " 0 " expression.ISIS 403739 demonstrates mice PCSK9mRNA under the dosage of 30nM and Geng Gao dose dependent descends.ISIS 403740 shows the decline of mice PCSK9mRNA under two maximum dose levels.

Table 77: comprise the Antisense Suppression of the short antisense compounds of BNAs to mice PCSK9

People Hep3B cell is handled with the short antisense compounds of nM concentration as herein described.After the processing time,, as described herein by quantitative PCR in real time measurement PCSK9mRNA level from cell separation RNA.The PCSK9mRNA level is normalized to cyclophilin mRNA level, and back one level is to measure by PCR in real time with cyclophilin Auele Specific Primer probe groups.The result suppresses percent with the PCSK9 with respect to untreatment control cell and represents.Data show in table 78, prove with ISIS 403740 and handle the reduction that dose dependent appears in descendant PCSK9mRNA.ISIS 403739 demonstrates dose dependent and descends under higher dosage.

Table 78: comprise the Antisense Suppression of the short antisense compounds of BNAs to mice PCSK9

The Antisense Suppression of GCGR in the embodiment 21:HepG2 cell

Tested of the effect of the short antisense compounds of targeting GCGR nucleic acid to external GCGR mRNA.

The HepG2 cell

The cultivation HepG2 cell that with the density in 96 orifice plates is 10000 cells/well is handled by the antisense oligonucleotide of usefulness 25,50,100 described herein or 200nM.After the processing time,, as described herein by quantitative PCR in real time measurement GCGRmRNA level from cell separation RNA.With GCGR mRNA level according to passing through

Total rna content of measuring is adjusted.The result reduces percent with the GCGRmRNA with respect to untreatment control cell and represents.

Table 79 has provided with the data after ISIS 327161 (a kind of 3-10-3MOE gapmer) processing of prescribed dose.ISIS 327161 has reduced GCGR mRNA in the dose dependent mode.

Table 79: short antisense compounds is to the Antisense Suppression of the GCGR in the HepG2 cell

ISIS NO.	Seq ID NO	Sequence (5 '-3 ')	The Gapmer die body	25 nM	50 nM	100 nM	200 nM
								327161	520	AGCTGCTGTACATC	3-8-3 MOE	-36	-30	-33	-64

The monkey hepatocyte

Tested of the effect of the short antisense compounds of other targeting GCGR nucleic acid to external monkey GCGRmRNA.Former generation monkey hepatocyte usefulness 25,50,100 as described herein of cultivating or the short antisense compounds of 200nM are handled.After the processing time,, as described herein by quantitative PCR in real time measurement GCGR mRNA level from cell separation RNA.With GCGR mRNA level according to passing through

Total rna content of measuring is adjusted.The result reduces percent with the GCGR mRNA with respect to untreatment control cell and represents in table 80.

Table 80: short antisense compounds is to the Antisense Suppression of the GCGR in the former generation monkey hepatocyte

ISIS NO.	Seq ID NO	Sequence (5 '-3 ')	The Gapmer die body	25 nM	50 nM	100 nM	200 nM
								327131	489	ATGTTGGCCGTGGT	3-8-3 MOE	0	-8	-36	-36
327161	520	AGCTGCTGTACATC	3-8-3 MOE	-19	-33	-55	-54

Embodiment 22: short antisense compounds is to the Antisense Suppression of DGAT2

Tested of the effect of the short antisense compounds of targeting DGAT2 nucleic acid to external DGAT2mRNA.Handle cultivation A10 cell in 96 orifice plates with the short antisense compounds of 75mM.After the processing time,, as described herein by quantitative PCR in real time measurement DGAT2mRNA level from cell separation RNA.With the DGAT2mRNA level according to passing through

Total rna content of measuring is adjusted.The result suppresses percent with the DGAT2 with respect to untreatment control cell and represents in table 81.

The Antisense Suppression of DGAT2 in the table 81:A10 cell

ISIS NO.	Seq ID NO	Sequence (5 '-3 ')	The Gapmer die body	The % contrast
					372491	795	ACATGAGGATGACACT	3-10-3MOE	80
372500	702	GTGTGTCTTCACCAGC	3-10-3MOE	16
					372501	704	TTGTGTGTCTTCACCA	3-10-3MOE	28
372503	708	GCAGGTTGTGTGTCTT	3-10-3MOE	35
					372508	719	AGTTCCTGGTGGTCAG	3-10-3MOE	35
372516	805	TACAGAAGGCACCCAG	3-10-3MOE	27
					372524	738	GCCAGGCATGGAGCTC	3-10-3MOE	21
372530	746	TCGGCCCCAGGAGCCC	3-10-3MOE	35
					372546	825	TTGGTCTTGTGATTGT	3-10-3MOE	34
372563	691	AGCCAGGTGACAGA	2-10-2MOE	48
					372569	796	CATGAGGATGACAC	2-10-2MOE	104
372578	703	TGTGTCTTCACCAG	2-10-2MOE	59
					372580	707	GGTTGTGTGTCTTC	2-10-2MOE	48
372586	720	GTTCCTGGTGGTCA	2-10-2MOE	40
					372594	806	ACAGAAGGCACCCA	2-10-2MOE	77
372602	739	CCAGGCATGGAGCT	2-10-2MOE	39
					372618	765	GTGGTACAGGTCGA	2-10-2MOE	29
372624	826	TGGTCTTGTGATTG	2-10-2MOE	56

In external dose response experiment, tested the short antisense compounds of other targeting DGAT2mRNA.The A10 cell prepare as mentioned above and with 6.25,12.5,25.0,50.0,100.0 and the short antisense compounds of 200.0nM handle, to determine that DGAT2 suppresses whether to occur in the dose dependent mode.The short antisense compounds of each that provides in the digital proof table 82 all reduces rat DGAT2mRNA in the dose dependent mode.The result represents with the inhibition percent with respect to untreatment control cell." 0 " expression DGAT2mRNA does not reduce.

The dose-dependent inhibition of DGAT2 in the table 82:A10 cell

ISIS NO.	Seq ID NO	Sequence (5 '-3 ')	The Gapmer die body	6.25 nM	12.5 nM	25.0 nM	50.0 nM	100.0 nM	200.0 nM
										372562	784	GTCTTGGAGGGCCG	2-10-2 MOE	0	0	0	36	48	75

372568	794	GACACTGCAGGCCA	2-10-2 MOE	0	0	15	26	72	69
										372586	720	GTTCCTGGTGGTCA	2-10-2 MOE	19	0	7	22	45	77
372602	739	CCAGGCATGGAGCT	2-10-2 MOE	0	0	0	18	47	76
										372618	765	GTGGTACAGGTCGA	2-10-2 MOE	0	5	0	27	65	80

At testing in vitro the short antisense compounds of other targeting DGAT2mRNA.The A10 cell prepare as mentioned above and with 0.62,1.85,5.56,16.67,50.0 and the short antisense compounds of 150.0nM handle, to determine that DGAT2 suppresses whether to occur in the dose dependent mode.The quantitative PCR in real time of DGAT2mRNA usefulness as described herein is measured.The short antisense compounds of each that provides in the digital proof following table 83 all suppresses rat DGAT2mRNA in the dose dependent mode.The result suppresses percent with the rat DGAT2 with respect to untreatment control cell and represents.Wherein the decline of DGAT2mRNA is not observed in " 0 " expression.

The dose-dependent inhibition of DGAT2 in the table 83:A10 cell

ISIS NO.	Seq ID NO	Sequence (5 '-3 ')	The Gapmer die body	0.62 nM	1.85 nM	5.56 nM	16.67 nM	50 nM	150 nM
										372500	702	GTGTGTCTTCACCAGC	3-10-3 MOE	0	0	0	18	64	88
372501	704	TTGTGTGTCTTCACCA	3-10-3 MOE	1	5	10	11	25	68
										372503	708	GCAGGTTGTGTGTCTT	3-10-3 MOE	7	10	4	25	54	80
372508	719	AGTTCCTGGTGGTCAG	3-10-3 MOE	0	0	6	14	39	71
										372516	805	TACAGAAGGCACCCAG	3-10-3 MOE	1	10	0	4	35	81
372524	738	GCCAGGCATGGAGCTC	3-10-3 MOE	7	0	5	30	68	91
										372530	746	TCGGCCCCAGGAGCCC	3-10-3 MOE	0	2	0	10	38	78
372546	825	TTGGTCTTGTGATTGT	3-10-3 MOE	0	2	11	4	48	78
										372563	691	AGCCAGGTGACAGA	2-10-2 MOE	0	0	0	1	4	46
372578	703	TGTGTCTTCACCAG	2-10-2 MOE	0	0	0	2	7	42
										372580	707	GGTTGTGTGTCTTC	2-10-2 MOE	0	5	5	3	16	42
372586	720	GTTCCTGGTGGTCA	2-10-2 MOE	0	0	0	0	7	55

372594	806	ACAGAAGGCACCCA	2-10-2 MOE	0	0	0	0	2	15
										372602	739	CCAGGCATGGAGCT	2-10-2 MOE	0	0	10	0	19	51
372618	765	GTGGTACAGGTCGA	2-10-2 MOE	0	0	0	0	30	60
										372624	826	TGGTCTTGTGATTG	2-10-2 MOE	0	0	0	1	16	38

The Antisense Suppression of people PTP1B in the embodiment 23:HuVEC cell

Tested of the effect of the short antisense compounds of targeting PTP1B nucleic acid to external PTP1B mRNA.With the density in 96 orifice plates is that the cultivation HuVEC cell short antisense compounds with 3nM as described herein of 5000 cells/well is handled.After the processing time,, as described herein by quantitative PCR in real time measurement PTP1B mRNA level from cell separation RNA.With PTP1B mRNA level according to passing through

Total rna content of measuring is adjusted.The result suppresses percent (%Inhib) expression with the PTP1B with respect to untreatment control cell.Digital proof in the table 84, short antisense compounds targeting PTP1B nucleic acid, that have the 2-10-2Gapmer die body can suppress the PTP1B in the HuVEC cell.

Table 84: short antisense compounds is to the Antisense Suppression of the PTP1B in the HuVEC cell

ISIS NO.	SEQ ID NO	The Gapmer die body	％Inhib
				399301	1542	2-10-2OMe	55
404137	1053	2-10-2MOE	76
				404138	1054	2-10-2MOE	76
404139	1052	2-10-2MOE	80
				404140	1051	2-10-2MOE	73

The Antisense Suppression of people PTP1B in the embodiment 24:HepG2 cell

Tested of the effect of the short antisense compounds of targeting PTP1B nucleic acid to external PTP1B mRNA.With the density in 96 orifice plates is that the cultivation HepG2 cell of 10000 cells/well is handled with the antisense oligonucleotide of 25nM.After the processing time,, as described herein by quantitative PCR in real time measurement PTP1B mRNA level from cell separation RNA.With the PTP1BmRNA level according to passing through

Total rna content of measuring is adjusted.The result suppresses percent (%Inhib) expression with the PTP1B with respect to untreatment control cell.Digital proof in the table 85, short antisense compounds targeting PTP1B nucleic acid, that have the 2-10-2Gapmer die body can suppress the PTP1B in the HepG2 cell.

Table 85: short antisense compounds is to the Antisense Suppression of the PTP1B in the HepG2 cell

ISIS NO.	SEQ ID NO	The Gapmer die body	％Inhib
				399301	1542	2-10-2OMe	43
404137	1053	2-10-2MOE	71
				404138	1054	2-10-2MOE	86
404139	1052	2-10-2MOE	45
				404140	1051	2-10-2MOE	93

Embodiment 25: at the HuVEC cell: the Antisense Suppression of carrying out PTP1B in the dose response experiment

With the inoculation of the density of 5000 cells/well, and short antisense compounds with the nM concentration shown in the table 86 as described herein is handled with human vascular endothelial (HuVEC) cell.After the processing time,, as described herein by quantitative PCR in real time measurement PTP1B mRNA level from cell separation RNA.With PTP1B mRNA level according to passing through

Total rna content of measuring is adjusted.Measure the mRNA level with two kinds of different people PTP1B primer probe groups.The result of primer probe groups (PPS) 198 shows that in table 86 result of primer probe groups (PPS) 3000 shows in table 87.The result represents with the PTP1B mRNA expression inhibiting percent with respect to untreatment control cell.Wherein the decline of PTP1B mRNA is not observed in " 0 " expression.Shown in table 86 and 87, PTP1B mRNA level reduces in the dose dependent mode.

The dose response of people PTP1B in the table 86:HuVEC cell uses PPS 198

The dose response of people PTP1B in the table 87:HuVEC cell uses PPS 3000

Embodiment 26: short antisense compounds is to the Antisense Suppression of ApoB

Tested the interior effect of body of short antisense compounds shown in the table 88.Give 6 ages in week male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) continuous three weeks weekly twice intraperitoneal give 3.2,1,0.32 or the dosage of .1umol/kg.5-10-5MOE gapmer is as control treatment.Put to death mice the last time behind the dosage in about 48 hours.Collect liver organization and carry out the RNA separation, collect blood and carry out the serum chemistry analysis.ApoB mRNA level is as described herein to be measured by PCR in real time.ApoB mRNA level is normalized to the rna level of measuring by RIBOGREEN, and in table 89 to represent with respect to the inhibition percent of the ApoB mRNA level in the saline treatment control animal.

Table 88: the short antisense compounds of targeting ApoB nucleic acid

ISIS NO	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
				387462	GGTACATGGAAGTC	2-10-2 methylene oxygen base BNA	190
398296	GGTACATGGAAGTC	2-10-2 6 '-(S)-methyl methylene oxygen base BNA	190

Table 89: comprise the Antisense Suppression of the short antisense compounds of BNA to ApoB

IsisNo	Dosage (umol/kg)	％Inhib
			379818	1	56
387462	0.1	33
				0.32	57
	1	93
				3.2	99
398296	0.1	17
				0.32	35
	1	80
				3.2	98

Table 89 is presented at after having the 2-10-2Gapmer die body and be with the short antisense compounds processing of BNA modification in the pterion, and ApoB mRNA level reduces in the dose dependent mode.Under 1umol/kg dosage, short antisense compounds to the inhibition of ApoB greater than 5-10-5MOEgapmer viewed inhibition under suitable dosage.1 and the short antisense compounds dosage of 3.2umol/kg under, cholesterol has reduced.

Judge that by organ weight and body weight, serum transaminase, bilirubin, blood urea nitrogen and kreatinin short antisense compounds seldom shows and even do not show adverse side effect.

Embodiment 27: short antisense compounds is to the Antisense Suppression of PTEN

Tested the interior effect of body of short antisense compounds shown in the table 90.Give 6 ages in week male Balb/c mice (Jackson Laboratory, Bar Harbor, ME) continuous three weeks weekly twice intraperitoneal give 3.2,1,0.32 or the dosage of .1umol/kg.5-10-5MOE gapmer is as control treatment.Put to death mice the last time behind the dosage in about 48 hours.Collect liver organization and carry out the RNA separation, collect blood and carry out the serum chemistry analysis.PTEN mRNA level is as described herein to be measured by PCR in real time.PTEN mRNA level is normalized to the rna level of measuring by RIBOGREEN, and in table 91 to represent with respect to the inhibition percent of the PTEN mRNA level in the saline treatment control animal.

Table 90: the short antisense compounds of targeting PTEN nucleic acid

ISIS NO	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
				392063	AGGCCAGTGCTAAG	2-10-2 methylene oxygen base BNA	1226
392749	AGGCCAGTGCTAAG	2-10-2 (6 ' S)-6 '-methyl methylene oxygen base BNA	1226
				396006	AGGCCAGTGCTAAG	2-10-2 α-L-methylene oxygen base BNA	1226

Table 91: comprise the Antisense Suppression of the short antisense compounds of BNA modification to PTEN

IsisNo	Dosage (umol/kg)	％ Inhib
			116847	1	47
392063	0.1	26
				0.32	43
	1	74
				3.2	96
392749	0.1	17
				0.32	34
	1	64
				3.2	96
396006	0.1	20
				0.32	32
	1	67
				3.2	88

Table 91 is presented at after having the 2-10-2Gapmer die body and be with the short antisense compounds processing of BNA modification in the pterion, and PTEN mRNA level level reduces in the dose dependent mode.Under 1umol/kg dosage, short antisense compounds to the inhibition of PTEN greater than 5-10-5MOE gapmer viewed inhibition under suitable dosage.

Except the ISIS 392063 of maximum dose level, do not observe significantly improving of serum transaminase.Generally speaking, lack antisense compounds and seldom show and even do not show adverse side effect.

Embodiment 28: the single that comprises the short antisense compounds of BNA modification gives

Give 6 ages in week male Balb/c mice (ME) single intraperitoneal injection is lacked antisense compounds for Jackson Laboratory, Bar Harbor, and dosage is 8,4,2 or 1 μ mol/kg.The short antisense compounds that is tried is ISIS 387462 and ISIS 398296.Each dosage group has four mices.5-10-5MOE gapmer is as control treatment.Put to death mice the last time behind the dosage in about 48 hours.Collect liver organization and carry out the RNA separation, collect blood and carry out the serum chemistry analysis.ApoB mRNA level is as described herein to be measured by PCR in real time.ApoB mRNA level is normalized to the rna level of measuring by RIBOGREEN, and in table 92 to represent with respect to the inhibition percent of the ApoB mRNA level in the saline treatment control animal.

Table 92: comprise the Antisense Suppression of the short antisense compounds of BNA to ApoB

IsisNo	Dosage (umol/kg)	％Inhib
			379818	8	77
387462	8	99
				4	93
	2	81
				1	58
398296	8	97
				4	81
	2	54
				1	19

After table 92 was presented at the short antisense compounds that single has the 2-10-2Gapmer die body and band BNA modifies in the pterion, ApoB mRNA level reduced in the dose dependent mode.Under 8umol/kg dosage, short antisense compounds to the inhibition of ApoB greater than 5-10-5MOE gapmer viewed inhibition under suitable dosage.The ED of ISIS 387462₅₀Be 3.9mg/kg, the ED of ISIS 398296₅₀Be 8.7mg/kg.Cholesterol also reduces in the dose dependent mode.Triglyceride reduces under maximum dose level.

Judge that by organ weight and body weight, serum transaminase, bilirubin, blood urea nitrogen and kreatinin short antisense compounds seldom shows and even do not show adverse side effect.

In similarly single is studied, ISIS 392748 (has SEQ ID NO:1226, the 2-10-2Gapmer die body, wherein the nucleotide in pterion comprises (6 ' R)-6 '-methyl methylene oxygen base BNA modification) reduced PTEN mRNA in the dose dependent mode.In addition, and ISIS 392749 (have SEQ ID NO:1226, the 2-10-2Gapmer die body, wherein the nucleotide in pterion comprises (6 ' S)-6 '-methyl methylene oxygen base BNA modification) reduced PTEN mRNA in the dose dependent mode.Have the sequence of 2-10-2Gapmer die body, SEQ ID NO:1226 and the short antisense compounds that 6-(S)-CH2-O-CH3-BNA modifies, in similar body, also reduced PTEN mRNA in the research.Have the sequence of 2-10-2Gapmer die body, SEQ ID NO:1226 and the short antisense compounds that 6-(S)-CH2-O-CH3-BNA modifies, in similar body, also reduced PTEN mRNA in the research.

Embodiment 29: the single that comprises the short antisense compounds of BNA modification gives

Give 6 ages in week male Balb/c mice (ME) single intraperitoneal injection is lacked antisense compounds for Jackson Laboratory, Bar Harbor, and dosage is 8,4,2 or 1 μ mol/kg.Each dosage group has four mices.The chemical compound that is tried is ISIS 392063, ISIS 392749 and ISIS 366006.5-10-5MOE gapmer is as control treatment.Put to death mice the last time behind the dosage in about 48 hours.Collect liver organization and carry out the RNA separation, collect blood and carry out the serum chemistry analysis.ApoB mRNA level is as described herein to be measured by PCR in real time.ApoB mRNA level is normalized to the rna level of measuring by RIBOGREEN, and in table 93 to represent with respect to the inhibition percent of the ApoB mRNA level in the saline treatment control animal.

Table 93: comprise the Antisense Suppression of the short antisense compounds of BNA modification to PTEN

IsisNo	Dosage (umol/kg)	％ Inhib
			116847	8	62
392063	8	92
				4	82
	2	58
				1	38
396565	8	76
				4	38
	2	24
				1	11
396006	8	94
				4	82
	2	48
				1	18

Table 93 is presented at after having the 2-10-2Gapmer die body and be with the short antisense compounds processing of BNA modification in the pterion, and PTEN mRNA level reduces in the dose dependent mode.Under 8umol/kg dosage, short antisense compounds to the inhibition of PTEN greater than 5-10-5MOEgapmer viewed inhibition under suitable dosage.The estimation ED of ISIS 392063₅₀Be 7mg/kg, ISIS 396565 is 17.4mg/kg, and ISIS 396006 is 9.3mg/kg.

Except the ISIS 392063 of maximum dose level, do not observe significantly improving of serum transaminase.Generally speaking, lack antisense compounds and seldom show and even do not show adverse side effect.

Embodiment 30: comprise the Antisense Suppression of the short antisense compounds of Palmic acid conjugate to ApoB

Give 6 ages in week male Balb/c mice (ME) single intraperitoneal injection is lacked antisense compounds for Jackson Laboratory, Bar Harbor, and dosage is 2.5,1.0,0.4 and 0.16umol/kg.Each dosage group has four mices.The chemical compound that is tried shows in table 94.5-10-5MOEgapmer is as control treatment.Put to death mice the last time behind the dosage in about 48 hours.Collect liver organization and carry out the RNA separation, collect blood and carry out the serum chemistry analysis.The ApoBmRNA level is as described herein to be measured by PCR in real time.ApoB mRNA level is normalized to the rna level of measuring by RIBOGREEN, and in table 95 to represent with respect to the inhibition percent of the ApoB mRNA level in the saline treatment control animal.

Table 94: the short antisense compounds that comprises the Palmic acid conjugate

ISIS NO	Sequence (5 '-3 ')	The Gapmer die body	SEQ ID NO
				387462	GGTACATGGAAGTC	2-10-2 methylene oxygen base BNA	190
391871	GGTACATGGAAGTC	Cytosine not modified (i.e. the 2-10-2MOE of 2 '-(butyl acetamido)-palmitamide on 5 ' nucleotide subsitution) in 1-1-10-22 '-(butyl acetamido)-palmitamide/MOE/MOE interval	190
				391872	GGTACATGGAAGTC	Cytosine not modified (i.e. the 2-10-2 methylene oxygen base BNA of 2 '-(butyl acetamido)-palmitamide on 5 ' nucleotide subsitution) in 1-1-10-22 '-(butyl acetamido)-palmitamide/methylene oxygen base BNA/ methylene oxygen base BNA interval	190

Table 95: the Antisense Suppression that comprises the short antisense compounds of Palmic acid conjugate

IsisNo	Dosage (umol/kg)	％Inhib
			5-10-5	2.5	54
387462	2.5	99
				1.0	91
	0.4	65
				0.16	16
391871	2.5	49
				1.0	18
	0.4	5
				0.16	0
391872	2.5	99
				1.0	92
	0.4	50
				0.16	18

Table 95 is presented at after the short antisense compounds processing with Palmic acid (C16) conjugate, and ApoB mRNA level reduces in the dose dependent mode.Under 2.5umol/kg dosage, short antisense compounds to the inhibition of ApoB greater than 5-10-5MOE gapmer viewed inhibition under suitable dosage.In this research, the estimation ED of ISIS 387462₅₀Be 1.5mg/kg, ISIS 391871 is 13.1mg/kg, and ISIS 391872 is 1.9mg/kg.The estimation E`D of 5-10-5MOEgapmer₅₀Be 17.4mg/kg.Triglyceride 2.5 and the ISIS 387462 and the ISIS of 1.0mg/kg dosage lower for 391872 times.ISIS 387462 and ISIS 391872 are with the obvious hypercholesterolemia reducing of dose dependent mode, HDL-C and LDL-C; The decline of LDL-C is very obvious, so that it drops under the detectable limit.Generally speaking, lack antisense compounds and seldom show and even do not show adverse side effect.

Embodiment 31: comprise the Antisense Suppression of the short antisense compounds of BNA modification to PCSK9 in the body

Give 6 ages in week male Balb/c mice (ME) single intraperitoneal injection gives ISIS 403739 or 403740 short antisense compounds for Jackson Laboratory, Bar Harbor, and dosage is 15,4.7,1.5 and .47umol/kg.Each dosage group has four mices.5-10-5MOE gapmer is as control treatment.Put to death mice the last time behind the dosage in about 72 hours.Collect liver organization and carry out the RNA separation, collect blood and carry out the serum chemistry analysis.The PCSK9mRNA level is as described herein to be measured by PCR in real time.The PCSK9mRNA level is normalized to the cyclophilin mRNA level of measuring by PCR in real time.ISIS 403739 has reduced PCSK9mRNA about 70% with respect to saline control.It is about 13% that ISIS 403740 has reduced PCSK9 with respect to saline control, but this reduction is not significant on the statistics.Lower dosage does not lower PCSK9mRNA significantly.Generally speaking, lack antisense compounds and seldom show and even do not show adverse side effect.

Claims (37)

1. length is 8-16 monomeric short antisense compounds, it comprises 2 '-deoxyribonucleotide interval region, these interval region both sides are connected to the pterion, and wherein each pterion independently comprises 1-3 the monomer of modifying through high-affinity, and the nucleotide of wherein said short antisense compounds targeting coding GCCR.

2. the short antisense compounds of claim 1, the wherein said monomer of modifying through high-affinity is through sugar-modified nucleotide.

3. the short antisense compounds of claim 2, wherein at least one comprises abutment between 4 ' and the 2 ' position of sugar-modified nucleotide at sugar.

4. the short antisense compounds of claim 2, wherein each described nucleotide of modifying through high-affinity is given the Δ Tm of every nucleotide 1-4 degree.

5. the short antisense compounds of claim 2, wherein each describedly comprises 2 '-substituent group except that H or OH through sugar-modified nucleotide.

6. the short antisense compounds of claim 5, wherein at least one described be the dicyclic ring thuja acid of 4 '-2 ' bridge joint through sugar-modified nucleotide.

7. the short antisense compounds of claim 5, wherein each 2 '-substituent group independently is alkoxyl, substituted alkoxy or halogen.

8. the short antisense compounds of claim 7, wherein each 2 '-substituent group is OCH₂CH₂OCH₃

9. the short antisense compounds of claim 3, wherein each described conformation through sugar-modified nucleotide independently is β-D or α-L.

10. the short antisense compounds of claim 5, wherein each described abutment independently comprise 1 or 2-4 independently be selected from-[C (R₁) (R₂)]_n-,-C (R₁)=C (R₂)-,-C (R₁)=N-,-C (=NR₁)-,-C (=O)-,-C (=S)-,-O-,-Si (R₁)₂-,-S (=O)_x-and-N (R₁)-linking group;

Wherein

X is 0,1 or 2;

N is 1,2,3 or 4;

Each R₁And R₂Independent is H, protecting group, hydroxyl, C₁-C₁₂Alkyl, substituted C₁-C₁₂Alkyl, C₂-C₁₂Thiazolinyl, substituted C₂-C₁₂Thiazolinyl, C₂-C₁₂Alkynyl, substituted C₂-C₁₂Alkynyl, C₅-C₂₀Aryl, substituted C₅-C₂₀Aryl, heterocyclic radical, substituted heterocyclic radical, heteroaryl, substituted heteroaryl, C₅-C₇Alcyl, substituted C₅-C₇Alcyl, halogen, OJ₁, NJ₁J₂, SJ₁, N₃, COOJ₁, acyl group (C (=O)-H), substituted acyl group, CN, sulfonyl (S (=O)₂-J₁) or sulfinyl (S (=O)-J₁); With

Each J₁And J₂Independent is H, C₁-C₁₂Alkyl, substituted C₁-C₁₂Alkyl, C₂-C₁₂Thiazolinyl, substituted C₂-C₁₂Thiazolinyl, C₂-C₁₂Alkynyl, substituted C₂-C₁₂Alkynyl, C₅-C₂₀Aryl, substituted C₅-C₂₀Aryl, acyl group (C (=O)-H), substituted acyl group, heterocyclic radical, substituted heterocyclic radical, C₁-C₁₂Aminoalkyl, substituted C₁-C₁₂Aminoalkyl or protecting group.

11. the short antisense compounds of claim 10, wherein each described abutment independently is 4 '-CH₂-2 ', 4 '-(CH₂)₂-2 ', 4 '-CH₂-O-2 ', 4 '-(CH₂)₂-O-2 ', 4 '-CH₂-O-N (R₁)-2 ' and 4 '-CH₂-N (R₁)-O-2 '-, each R wherein₁Independent is H, protecting group or C₁-C₁₂Alkyl.

12. the short antisense compounds of claim 1, wherein each described monomer of modifying through high-affinity independently is selected from dicyclic ring thuja acid or other nucleotide through 2 '-modification.

13. the short antisense compounds of claim 12, wherein said nucleotide through 2 '-modification is selected from halogen, pi-allyl, amino, azido, thio group, O-pi-allyl, O-C₁-C₁₀Alkyl ,-OCF₃, O-(CH₂)₂-O-CH₃, 2 '-O (CH₂)₂SCH₃, O-(CH₂)₂-O-N (R_m) (R_n) or O-CH₂-C (=O)-N (R_m) (R_n), each R wherein_mAnd R_nIndependent is H or substituted or unsubstituted C₁-C₁₀Alkyl.

14. the short antisense compounds of claim 13, wherein said nucleotide through 2 '-modification is 2 '-OCH₂CH₂OCH₃Nucleotide.

15. the short antisense compounds of claim 1, wherein at least one Dan Julian key is modified Dan Julian key.

16. the short antisense compounds of claim 15, wherein said modified Dan Julian key is that thiophosphate connects key.

17. the short antisense compounds of claim 1, wherein each Dan Julian key is to connect key between the thiophosphate nucleoside.

18. each short antisense compounds of claim 1-17, the length of described short antisense compounds is 8-15 monomer.

19. the short antisense compounds of claim 18, the length of described short antisense compounds are 9-15 monomer.

20. the short antisense compounds of claim 18, the length of described short antisense compounds are 10-15 monomer.

21. the short antisense compounds of claim 18, the length of described short antisense compounds are 9-14 monomer.

22. the short antisense compounds of claim 18, the length of described short antisense compounds are 10-14 monomer.

23. the short antisense compounds of claim 18, the length of described short antisense compounds are 9-13 monomer.

24. the short antisense compounds of claim 18, the length of described short antisense compounds are 10-13 monomer.

25. the short antisense compounds of claim 18, the length of described short antisense compounds are 9-12 monomer.

26. the short antisense compounds of claim 18, the length of described short antisense compounds are 10-12 monomer.

27. the short antisense compounds of claim 18, the length of described short antisense compounds are 9-11 monomer.

28. the short antisense compounds of claim 18, the length of described short antisense compounds are 10-11 monomer.

29. the short antisense compounds of claim 18, the length of described short antisense compounds are 8 monomers.

30. the short antisense compounds of claim 18, the length of described short antisense compounds are 9 monomers.

31. the short antisense compounds of claim 18, the length of described short antisense compounds are 10 monomers.

32. the short antisense compounds of claim 18, the length of described short antisense compounds are 11 monomers.

33. the short antisense compounds of claim 18, the length of described short antisense compounds are 12 monomers.

34. the short antisense compounds of claim 18, the length of described short antisense compounds are 13 monomers.

35. the short antisense compounds of claim 18, the length of described short antisense compounds are 14 monomers.

36. the short antisense compounds of claim 18, the length of described short antisense compounds are 15 monomers.

37. the short antisense compounds of claim 18, the length of described short antisense compounds are 16 monomers.