The present application is a divisional application of chinese patent application having an application date of 27/2/2009, an application number of 200980111248.2, and an invention name of "amide compound, composition, and use thereof".
Disclosure of Invention
Provided herein are biphenyl and pyridylphenylamide compounds and pharmaceutical compositions thereof having efficacy and selectivity for the prevention and treatment of diseases associated with neurological and inflammatory disorders and dysfunctions.
In particular, the compounds, pharmaceutical compositions and methods provided are useful for treating, preventing or ameliorating a range of mammalian diseases, such as, but not limited to, pain of various origins or etiologies, such as acute, chronic, inflammatory and neuropathic pain, dental pain and headache (e.g., migraine, cluster headache and tension headache). In some embodiments, the compounds, pharmaceutical compositions and methods provided are useful for treating inflammatory pain and associated hyperalgesia and allodynia. In some embodiments, compounds, pharmaceutical compositions and methods are provided that are useful for treating neuropathic pain and related hyperalgesia and allodynia (e.g., trigeminal or herpetic neuralgia, diabetic neuropathy, causalgia, sympathetically maintained pain, and afferent nerve block syndromes, such as brachial plexus avulsion (avulsion)). In some embodiments, compounds, pharmaceutical compositions and methods are provided for use as anti-inflammatory agents for the treatment of arthritis and for the treatment of parkinson's disease, alzheimer's disease, asthma, myocardial infarction, neurodegenerative diseases, inflammatory bowel disease and autoimmune diseases, renal diseases, obesity, eating disorders, cancer, schizophrenia, epilepsy, sleep disorders, cognitive disorders, depression, anxiety, blood pressure and lipid metabolism disorders.
Thus, in one aspect, there is provided a compound having formula 1:
wherein
Each A, B and W is independently selected from CR4；
X' is selected from CR4aAnd N;
l is-C (R)2aR2b)-；
R1Selected from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, hydroxy C1-C4Alkyl radical, C3-C7cycloalkyl-C1-C4Alkyl or 4-7 membered heterocycloalkyl-C1-C4An alkyl group;
R2aand R2bEach independentlySelected from hydrogen, C1-C4Alkyl or hydroxy C1-C4An alkyl group;
R3is substituted or unsubstituted C1-C6An alkyl group; CH (OH) R3a、OR3a、CN、COR3a、COOR3a、SOR3a、SO2R3a、CONR3aR3b、SONR3aR3bOr SO2NR3aR3b；
R3aIs H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
R3bis H, substituted or unsubstituted C1-C6An alkyl group; or R3aAnd R3bTogether form a cycloheteroalkyl ring of 3-7 atoms;
R4each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted acyl, substituted or unsubstituted acylamino, substituted or unsubstituted alkylamino, substituted or unsubstituted alkylthio, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted alkylarylamino, substituted or unsubstituted amino, substituted or unsubstituted arylalkyl, sulfo, substituted sulfonyl, substituted sulfinyl, substituted sulfanyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted arylsulfonyl, azido, substituted or unsubstituted carbamoyl, carboxy, cyano, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted dialkylamino, halogen, nitro and mercapto;
R4aAnd R4bEach independently selected from H, substituted or unsubstituted alkyl, substituted or unsubstituted acyl, substituted or unsubstituted acylamino, substituted or unsubstituted alkylamino, substituted or unsubstituted alkylthio, substituted or unsubstituted alkoxy, orSubstituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted alkylarylamino, substituted or unsubstituted arylalkoxy, substituted or unsubstituted amino, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, sulfo, substituted sulfonyl, substituted sulfinyl, substituted sulfanyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted arylsulfonyl, azido, substituted or unsubstituted carbamoyl, carboxy, cyano, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloheteroalkyl, substituted or unsubstituted dialkylamino, halogen, heteroaryloxy, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroalkyl, hydroxy, nitro, and mercapto;
and subscript m' is selected from 0 to 4;
provided that
When R is3Is CO2Me、SO2Ph OR OR3aWhen R is1Is not unsubstituted phenyl;
When R is3Is SO2- (4-methylpiperazin-1-yl) or SO2- (thiomorpholin-1-yl) and X' is CH, R4bIs not H;
when R is3When Me is methyl substituted by alkoxy, R1Is not a substituted phenyl group;
when R is3Is CO2When H, R4bIs Cl, F, Br, Me, Et, OMe or CF3；
When X' is CR4a，R3Is CONR3aR3bAnd R is3aWhen is H, R3bIs not substituted n-pentyl, substituted pentynyl, substituted benzyl, substituted phenethyl, substituted thienylethyl or substituted thiazolylethyl; and
when R is1Is a 5-6 membered heterocycloalkylmethyl group, and R3Is CO2Me or n-Pr, R4bIs not Cl or 4-F;
or a pharmaceutically acceptable salt, N-oxide, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In a particular embodiment of formula 1, A, B and W are each CH.
In one embodiment of formula 1, L is selected from-CH2-、-CHMe-、-CMe2-、-CH(CH2OH) -and-CH (CH)2CH2OH)-。
In one embodiment of formula 1, L is selected from-CH2-and-CHMe-.
In one embodiment of formula 1, the compound is a compound of formula 2a, 2b, 2c, 2d, 2e, 2f, 2g, 2h, or 2 i:
wherein
X’、R1、R3a、R3b、R4a、R4bAnd m' is as described for formula 1; r2aIs H, Me, CH2OH or CH2CH2OH，
Or a pharmaceutically acceptable salt, solvate, N-oxide, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In another aspect, pharmaceutical compositions are provided comprising biphenyl and pyridylphenylamide provided herein and a pharmaceutically acceptable carrier, excipient, or diluent. The pharmaceutical composition may comprise one or more compounds described herein.
It is to be understood that the compounds provided herein for use in pharmaceutical compositions and methods of treatment disclosed herein may be prepared to be pharmaceutically acceptable and used.
In another aspect, there is provided a method for preventing, treating or ameliorating a disease such as those mentioned herein, which may be particularly associated with, for example, arthritis, asthma, myocardial infarction, lipid metabolism disorders, cognitive disorders, anxiety, schizophrenia, depression, memory dysfunction, for example alzheimer's disease, inflammatory bowel disease and autoimmune diseases, comprising administering to a mammal in need thereof one or more compounds provided herein or pharmaceutical compositions thereof in an amount effective to prevent, treat or ameliorate said disease.
In another aspect, methods are provided for preventing, treating, or ameliorating a disease that results in a painful response or is associated with maintaining an imbalance in the basal activity of the sensory nerves in a mammal. The compounds provided herein are useful as analgesics for the treatment of pain of various origins or etiologies, such as acute pain, inflammatory pain (e.g., pain associated with osteoarthritis and rheumatoid arthritis); various neuropathic pain syndromes (e.g., post-herpetic neuralgia, trigeminal neuralgia, reflex sympathetic dystrophy (reflex sympathic dystrophy), diabetic neuropathy, Guillain-Barr é syndrome, fibromyalgia (fibromyalgia), phantom limb pain, post-mastectomy pain, polyneuropathy (perpheral neuropathy), HIV neuropathy, and chemotherapy-induced neuropathy and other iatrogenic neuropathies); visceral pain (e.g., visceral pain associated with gastroesophageal reflux disease (gastroesophageal reflux disease), irritable bowel syndrome, inflammatory bowel disease, pancreatitis, and various gynecological and urological conditions), dental pain, and headache (e.g., migraine, cluster headache, and tension headache).
In one aspect, a method for preventing, treating or ameliorating a neurodegenerative disease or disorder in a mammal is provided. The neurodegenerative disease or disorder can be, for example, parkinson's disease, alzheimer's disease, and multiple sclerosis; diseases and disorders mediated by or resulting in neuroinflammation, such as encephalitis; centrally mediated neuropsychiatric diseases and disorders such as depression, mania, bipolar disorder, anxiety, schizophrenia, eating disorders, sleep disorders, and cognitive disorders; epilepsy and seizures; prostate, bladder and bowel dysfunction, such as urinary incontinence, urinary hesitancy, rectal hypersensitivity, fecal incontinence, benign prostatic hypertrophy and inflammatory bowel disease; respiratory and airway diseases and disorders such as allergic rhinitis, asthma and reactive airway disease and chronic obstructive pulmonary disease; diseases and disorders mediated by or resulting in inflammation, such as rheumatoid arthritis and osteoarthritis, myocardial infarction, various autoimmune diseases and disorders; itch (itch)/pruritus, such as psoriasis; obesity; disorders of lipid metabolism; cancer; and renal diseases. Typically, the methods comprise administering to a mammal in need thereof one or more compounds provided herein or a pharmaceutical composition thereof in an amount effective to treat or prevent a disease.
In addition to the methods of treatment described above, the invention extends to the use of any compound of the invention in the manufacture of a medicament or a medicament which can be administered to such a treatment and the use of such a compound in the treatments disclosed and specified.
In other aspects, methods are provided for synthesizing the compounds described herein, wherein representative synthetic schemes and routes are described below. In some embodiments, methods of preparing enantiomerically pure compounds of formula 1 by asymmetric synthesis are provided. In some embodiments, methods of preparing enantiomerically pure compounds of formula 1 by chiral resolution are provided.
Other objects and advantages will be apparent to those skilled in the art in view of the following detailed description.
Detailed Description
Definition of
It is intended that the following terms have the meanings provided below and are used in understanding the specification and the intended scope of the invention.
When describing the present invention, which may include compounds, pharmaceutical compositions comprising such compounds, and methods of using such compounds and compositions, the following terms, if any, have the following meanings, unless otherwise specified. It is also to be understood that any of the moieties defined below may be substituted with various substituents when described herein, and that the corresponding definitions are intended to include such substituted moieties within the scope as described below. Unless otherwise stated, the term "substituted" is as defined below. It is also to be understood that the terms "group" and "radical" are considered herein to be used interchangeably.
The articles "a" and "an" are used herein to mean one or more (i.e., at least one) of the grammatical object of the article. By way of example, "analog" means one analog or more than one analog.
As defined herein, "acyl" or "alkanoyl" means the group-C (O) R20Wherein R is20Is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl radical, C3-C10Cycloalkylmethyl, 4-10 membered heterocycloalkyl, aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl. Representative examples include, but are not limited to, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl and benzylcarbonyl. Typical "acyl" groups are-C (O) H, -C (O) -C1-C8Alkyl, -C (O) - (CH)2)t(C6-C10Aryl), -C (O) - (CH)2)t(5-10 membered heteroaryl), -C (O) - (CH)2)t(C3-C10Cycloalkyl) and-C (O) - (CH)2)t(4-10 membered heterocycloalkyl) wherein t is an integer of 0 to 4.
"substituted acyl" or "substituted alkanoyl" means the group-C (O) R21Wherein R is21Independently are:
c substituted by halogen or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylArylalkyl radicals, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"amido" means the group-NR22C(O)R23Wherein R is22Is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, arylalkyl, 5-to 10-membered heteroaryl or heteroarylalkyl, R23Is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, arylalkyl, 5-10 membered heteroaryl, or heteroarylalkyl, as defined herein. Typical "acylamino" groups include, but are not limited to, formylamino, acetylamino, cyclohexylcarbonylamino, cyclohexylmethyl-carbonylamino, benzoylamino and benzylcarbonylamino. A specific exemplary "amido" group is-NR24C(O)-C1-C8Alkyl, -NR24C(O)-(CH2)t(C6-C10Aryl), -NR-24C(O)-(CH2)t(5-10 membered heteroaryl), -NR24C(O)-(CH2)t(C3-C10Cycloalkyl) and-NR24C(O)-(CH2)t(4-10 membered heterocycloalkyl) wherein t is an integer of 0 to 4, R24Each independently represents H or C1-C8An alkyl group.
"substituted amido" means the group-NR25C(O)R26Wherein:
R25independently is
H, C substituted by halogen or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-to 10-membered heterocycloalkyl、C6-C10Aryl, arylalkyl, 5-to 10-membered heteroaryl or heteroarylalkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy substitution; and
R26independently is
H, C substituted by halogen or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, arylalkyl, 5-to 10-membered heteroaryl or heteroarylalkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy substitution;
provided that R is25And R26Is not H.
"acyloxy" means the group-OC (O) R27Wherein R is27Is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl radical, C3-C10Cycloalkylmethyl, 4-10 membered heterocycloalkyl, aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl, as defined herein. Representative examples include, but are not limited to, formyl, acetyl, cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl and benzylcarbonyl. Typical "acyl" groups are-C (O) H, -C (O) -C1-C8Alkyl, -C (O) - (CH)2)t(C6-C10Aryl), -C (O) - (CH)2)t(5-10 membered heteroaryl), -C (O) - (CH)2)t(C3-C10Cycloalkyl) and-C (O)(CH2)t(4-10 membered heterocycloalkyl) wherein t is an integer of 0 to 4.
"substituted acyloxy" means the group-OC (O) R28Wherein R is28Independently is
C substituted by halogen or hydroxy1-C8An alkyl group; or
C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, arylalkyl, 5-to 10-membered heteroaryl or heteroarylalkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"alkoxy" means the group-OR29Wherein R is29Is C1-C8An alkyl group. Specific alkoxy groups are methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, tert-butoxy, sec-butoxy, n-pentoxy, n-hexoxy and 1, 2-dimethylbutoxy. Specific alkoxy groups are lower alkoxy groups having 1 to 6 carbon atoms. Other specific alkoxy groups have 1 to 4 carbon atoms.
"substituted alkoxy" means an alkoxy group substituted with one or more groups recited in the definition of "substituted" herein, and particularly means an alkoxy group having 1 or more substituents (e.g., 1 to 5 substituents, particularly 1 to 3 substituents, particularly 1 substituent) selected from amino, substituted amino, C6-C10Aryl, aryloxy, carboxy, cyano, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, halogen, 5-10 membered heteroaryl, hydroxy, nitro, thioalkoxy, thioaryloxy, mercapto, alkyl-S (O) -, aryl-S (O) -, alkyl-S (O)2And aryl-S (O)2-. A typical "substituted alkoxy" is-O- (CH)2)t(C6-C10Aryl), -O- (CH)2)t(5-10 membered heteroaryl), -O- (CH)2)t(C3-C10Cycloalkyl) and-O- (CH)2)t(4-10 membered heterocycloalkyl) wherein t is an integer from 0 to 4 and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl present may itself be unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy. A specific exemplary "substituted alkoxy" group is OCF3、OCH2CF3、OCH2Ph、OCH2-cyclopropyl, OCH2CH2OH and OCH2CH2NMe2。
"alkoxycarbonyl" means a radical-C (O) -OR30Wherein R is30Is represented by C1-C8Alkyl radical, C3-C10Cycloalkyl radical, C3-C10Cycloalkylalkyl, 4-10 membered heterocycloalkylalkyl, aralkyl, or 5-10 membered heteroarylalkyl, as defined herein. Typical "alkoxycarbonyl" groups are C (O) O-C1-C8Alkyl, -C (O) O- (CH)2)t(C6-C10Aryl), -C (O) O- (CH)2)t(5-10 membered heteroaryl), -C (O) O- (CH)2)t(C3-C10Cycloalkyl) and-C (O) O- (CH)2)t(4-10 membered heterocycloalkyl) wherein t is an integer of 1 to 4.
"substituted alkoxycarbonyl" means a radical-C (O) -OR31Wherein R is31Represents:
C1-C8alkyl radical, C3-C10Cycloalkyl radical, C3-C10Cycloalkylalkyl or 4-10 membered heterocycloalkylalkyl each substituted with halogen, substituted or unsubstituted amino or hydroxy; or
C6-C10Aralkyl or 5-to 10-membered heteroarylalkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"Aryloxycarbonyl" means the group-C (O) -OR32Wherein R is32Is represented by C6-C10Aryl, as defined herein. A typical "aryloxycarbonyl" group is-C (O) O- (C)6-C10Aryl).
"substituted aryloxycarbonyl" means the group-C (O) -OR33Wherein R is33To represent
C6-C10Aryl radical, unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"Heteroaryloxycarbonyl" means the group-C (O) -OR34Wherein R is34Represents a 5-10 membered heteroaryl group as defined herein. A typical "aryloxycarbonyl" group is-C (O) O- (5-10 membered heteroaryl).
"substituted heteroaryloxycarbonyl" means the group-C (O) -OR35Wherein R is35Represents:
5-10 membered heteroaryl, unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"Alkoxycarbonylamino" means a radical-NR36C(O)OR37Wherein R is36Is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl radical, C3-C10Cycloalkylmethyl, 4-10 membered heterocycloalkyl, aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl as defined herein, R37Is C1-C8Alkyl radical, C3-C10Cycloalkyl radical, C3-C10Cycloalkylmethyl, 4-10 membered heterocycloalkyl, aryl, arylalkyl, 5-10 membered heteroaryl or heteroarylalkyl as defined herein.
"alkyl" means a straight or branched chain aliphatic hydrocarbon having 1 to 20 carbon atoms. Specific alkyl groups have 1 to 12 carbon atoms. More specific alkyl groups are lower alkyl groups having 1 to 6 carbon atoms. Another specific alkyl group has 1-4 carbon atoms. Typical linear groups include methyl, ethyl, n-propyl and n-butyl. Branched means that one or more lower alkyl groups (e.g., methyl, ethyl, propyl, or butyl are attached to a linear alkyl chain, typical branching groups include isopropyl, iso-butyl, tert-butyl, and isopentyl.
"substituted alkyl" means an alkyl group as defined above substituted with one or more groups as described in the definition of "substituted" herein, and particularly means an alkyl group having 1 or more substituents (e.g., 1 to 5 substituents, particularly 1 to 3 substituents, particularly 1 substituent) selected from acyl, acylamino, acyloxy (-O-acyl or-OC (O) R)20) Alkoxy, alkoxycarbonyl, alkoxycarbonylamino (-NR' -alkoxycarbonyl OR-NH-C (O) -OR)27) Amino, substituted amino, aminocarbonyl (carbamoyl or amido or-C (O) -NR "2) Aminocarbonylamino (-NR' -C (O) -NR) "2) Aminocarbonyloxy (-O-C (O) -NR) "2) Aminosulfonyl, sulfonylamino, aryl, aryloxy, azido, carboxy, cyano, cycloalkyl, halogen, hydroxy, heteroaryl, nitro, mercapto, -S-alkyl, -S-aryl, -S (O) -alkyl, -S (O) -aryl, -S (O)2-alkyl and-S (O)2-an aryl group. In a toolIn embodiments of the invention, "substituted alkyl" means substituted with halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, -NR' "SO2R”、-SO2NR "R '", -C (O) R ", -C (O) OR", -OC (O) R ", -NR '" C (O) R ", -C (O) NR" R ' ", -NR" R ' "OR- (CR '" R ")mOR' "substituted C1-C8An alkyl group; wherein each R' is independently selected from H, C1-C8Alkyl, - (CH)2)t(C6-C10Aryl), - (CH)2)t(5-10 membered heteroaryl), - (CH)2)t(C3-C10Cycloalkyl) and- (CH)2)t(4-10 membered heterocycloalkyl) wherein t is an integer from 0 to 4 and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl present may itself be unsubstituted1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy. R' "and R" "each independently represent H or C1-C8An alkyl group.
"alkylene" means a divalent saturated alkylene group which may be straight or branched chain having from 1 to 11 carbon atoms and more specifically from 1 to 6 carbon atoms. The term is defined as, for example, methylene (-CH)2-) ethylene (-CH2CH2-), propylene isomers (e.g. -CH2CH2CH2-and-CH (CH)3)CH2-) etc. are typical.
"substituted alkylene" means a group as set forth in the definition of "substituted" herein, and particularly means an alkylene having 1 or more substituents (e.g., 1 to 5 substituents, particularly 1 to 3 substituents) selected from acyl, amido, acyl, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, amino-carbonylamino, aminocarbonyloxy, aryl, aryloxy, azido Nitrogen, carboxyl, cyano, halogen, hydroxy, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, mercapto, alkyl-S (O) -, aryl-S (O) -, alkyl-S (O)2And aryl-S (O)2-。
"alkenyl" means a monovalent ethylenically unsaturated hydrocarbon radical having from 2 to 11 carbon atoms, specifically from 2 to 8 carbon atoms, and more specifically from 2 to 6 carbon atoms, which may be straight or branched chain and has at least 1 and specifically from 1 to 2 sites of ethylenic unsaturation. Specific alkenyl groups include vinyl (-CH ═ CH)2) N-propenyl (-CH)2CH＝CH2) Isopropenyl (-C (CH)3)＝CH2) Vinyl and substituted vinyl, and the like.
"substituted alkenyl" means a group as set forth in the definition of "substituted" herein, and particularly means alkenyl having 1 or more substituents (e.g., 1 to 5 substituents, particularly 1 to 3 substituents) selected from acyl, acylamino, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxy, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, mercapto, alkyl-S (O) -, aryl-S (O) -, alkyl-S (O) —2And aryl-S (O)2-。
"alkenylene" means a divalent ethylenically unsaturated hydrocarbon radical having up to about 11 carbon atoms, and more specifically 2 to 6 carbon atoms, which may be straight or branched chain and has at least 1 and specifically 1 to 2 sites of ethylenic unsaturation. The term is taken to mean, for example, ethenylene (-CH-), propenylene isomers (e.g., -CH- ═ CHCH)2-and-C (CH)3) CH-and-CH-C (CH)3) -) etc. are typical.
"alkynyl" means an acetylenically or acetylenically unsaturated hydrocarbon radical, in particularIs an acetylenically unsaturated hydrocarbyl group having 2 to 11 carbon atoms, and more specifically 2 to 6 carbon atoms, which may be linear or branched and has at least 1, and specifically 1 to 2, acetylenically unsaturated sites. Specific non-limiting examples of alkynyl groups include ethynyl, ethynyl (-C ≡ CH), propynyl (-CH)2C.ident.CH) and the like.
"substituted alkynyl" means a group as set forth in the definition of "substituted" herein, and particularly means alkynyl having 1 or more substituents (e.g., 1 to 5 substituents and particularly 1 to 3 substituents) selected from acyl, amido, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxy, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, mercapto, alkyl-S (O) -, aryl-S (O) -, alkyl-S (O)2And aryl-S (O)2-。
"amino" means the radical-NH2。
"substituted amino" means an amino group substituted with one or more groups described in the definition of "substituted" herein and specifically means the group-N (R)38)2Wherein R is38Each independently selected from:
hydrogen, C1-C8Alkyl radical, C6-C10Aryl, 5-10 membered heteroaryl, 4-10 membered heterocycloalkyl or C3-C10A cycloalkyl group; or
C substituted by halogen or hydroxy1-C8An alkyl group; or
·-(CH2)t(C6-C10Aryl), - (CH)2)t(5-10 membered heteroaryl), - (CH)2)t(C3-C10Cycloalkyl) or- (CH)2)t(4-10 yuan)Heterocycloalkyl) in which t is an integer from 0 to 8, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy substitution; or
Two R38The groups together form an alkylene group.
When two R are38When the radicals are all hydrogen, -N (R)38)2Is an amino group. A typical "substituted amino" group is-NR39-C1-C8Alkyl, -NR39-(CH2)t(C6-C10Aryl), -NR-39-(CH2)t(5-10 membered heteroaryl), -NR39-(CH2)t(C3-C10Cycloalkyl) and-NR39-(CH2)t(4-10 membered heterocycloalkyl) wherein t is an integer of 0 to 4, R39Each independently represents H or C1-C8An alkyl group; and any alkyl groups present may themselves be substituted by halogen, substituted or unsubstituted amino or hydroxy; and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl radicals present may themselves be unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy. For the avoidance of doubt, the term "substituted amino" includes the groups alkylamino, substituted alkylamino, alkylarylamino, substituted alkylarylamino, arylamino, substituted arylamino, dialkylamino and substituted dialkylamino as defined above.
"alkylamino" means the group-NHR40Wherein R is40Is C1-C8An alkyl group.
"substituted alkylamino" means the radical-NHR41Wherein R is41Is C1-C8An alkyl group; and alkyl is substituted by halogen, substituted or unsubstituted amino, hydroxy, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, 5-10 membered heteroaryl, aralkyl or heteroaralkyl substitution; and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl radicals present may themselves be unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"Alkylarylamino" means the group-NR42R43Wherein R is42Is aryl, R43Is C1-C8An alkyl group.
"substituted alkylarylamino" means the group-NR44R45Wherein R is44Is aryl, R45Is C1-C8An alkyl group; alkyl substituted by halogen, substituted or unsubstituted amino, hydroxy, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, 5-10 membered heteroaryl, aralkyl or heteroaralkyl substitution; and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl radicals present may themselves be unsubstituted C1-C4Alkyl, halogen, cyano, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"arylamino" means the group-NHR46Wherein R is46Is selected from C6-C10Aryl and 5-10 membered heteroaryl, as defined herein.
"substituted arylamino" means the group-NHR47Wherein R is47Independently selected from C6-C10Aryl and 5-10 membered heteroaryl; c in which any aryl or heteroaryl radical present may itself be unsubstituted1-C4Alkyl, halogen, cyano, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"dialkylamino" means a radical-NR48R49Wherein R is48And R49Each independently selected from C1-C8An alkyl group.
"substituted dialkylamino" means the group-NR50R51Wherein each R59And R51Independently selected from C1-C8An alkyl group; at least one alkyl group being independently substituted by halogen, hydroxy, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, 5-10 membered heteroaryl, aralkyl or heteroaralkyl substitution; any aryl, heteroaryl, cycloalkyl or heterocycloalkyl radicals present may themselves be unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"Diarylamino" means the radical-NR52R53Wherein R is52And R53Each independently selected from C6-C10And (4) an aryl group.
"aminosulfonyl" or "sulfonamide" means the group-S (O)2)NH2。
"substituted aminosulfonyl" or "substituted sulfonamide" means a group such as-S (O)2)N(R54)2Wherein R is54Each independently selected from:
·H、C1-C8alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-10 membered heteroaryl and heteroaralkyl; or
C substituted by halogen or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-to 10-membered heteroaryl or heteroaralkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy substitution;
provided that at least one R54Is not H.
A typical "substituted aminosulfonyl" or "substituted sulfonamide" group is-S (O)2)N(R55)-C1-C8Alkyl, -S (O)2)N(R55)-(CH2)t(C6-C10Aryl group, -S (O)2)N(R55)-(CH2)t(5-10 membered heteroaryl), -S (O)2)N(R55)-(CH2)t(C3-C10Cycloalkyl) and-S (O)2)N(R55)-(CH2)t(4-10 membered heterocycloalkyl) wherein t is an integer from 0 to 4; r55Each independently represents H or C1-C8An alkyl group; any aryl, heteroaryl, cycloalkyl or heterocycloalkyl radicals present may themselves be unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"aralkyl" or "arylalkyl" means an alkyl group as defined above substituted with one or more aryl groups as defined above. A particular aralkyl or arylalkyl group is an alkyl group substituted with one aryl group.
"substituted aralkyl" or "substituted arylalkyl" means an alkyl group as defined above substituted with one or more aryl groups; c wherein at least one of the aryl groups present may itself be unsubstituted1-C4Alkyl, halogen, cyano, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"aryl" means a monovalent aromatic hydrocarbon radical derived by the removal of one hydrogen atom from a single carbon atom of a parent aromatic ring system. Specifically, aryl means an aromatic monocyclic or polycyclic ring structure comprising 5 to 12 ring members, more typically 6 to 10 ring members. When aryl is a monocyclic system, it preferably contains 6 carbon atoms. Typical aryl groups include, but are not limited to, groups derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, 5, coronene, fluoranthene, fluorene, hexacene, hexylene, hexalene, asymmetric indacene, symmetric indacene, indane, indene, naphthalene, octabenzene, octaphene, octalene, ovalene, penta-2, 4-diene, pentacene, pentalene, dibenzo [ b, h ] phenanthrene, perylene, phenalene, phenanthrene, benzo [ a, i ] phenanthrene, heptadiene, pyrene, pyranthrene, rubicene, benzo (9,10) phenanthrene, and binaphthylene (trinaphthallene). Specific aryl groups include phenyl, naphthyl, indenyl, and tetrahydronaphthyl.
"substituted aryl" means an aryl group substituted with one or more groups recited in the definition of "substituted" herein and specifically means an aryl group, which may be optionally substituted with 1 or more substituents (e.g., 1 to 5 substituents, specifically 1 to 3 substituents, specifically 1 substituent). In particular, "substituted aryl" means substituted by one or more groups selected from halogen, C1-C8Alkyl radical, C1-C8Haloalkyl groupCyano, hydroxy, C1-C8Alkoxy and amino groups.
Representative examples of substituted aryl groups include the following:
in these formulae, R56And R57One of which may be hydrogen, R56And R57At least one of each is independently selected from C1-C8Alkyl radical, C1-C8Haloalkyl, 4-to 10-membered heterocycloalkyl, alkanoyl, C1-C8Alkoxy, heteroaryloxy, alkylamino, arylamino, heteroarylamino, NR58COR59、NR58SOR59NR58SO2R59COOalkyl, COOaryl, CONR58R59、CONR58OR59、NR58R59、SO2NR58R59S-alkyl, SO2Alkyl, S aryl, SO2An aryl group; or R56And R57May be linked to form a 5-8 atom ring (saturated or unsaturated) optionally containing one or more heteroatoms selected from N, O or S. R60And R61Independently of each other is hydrogen, C1-C8Alkyl radical, C1-C4Haloalkyl, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, substituted aryl, 5-10 membered heteroaryl.
"fused aryl" means an aryl group having two ring carbons common to a second aryl ring or to an aliphatic ring.
"arylalkoxy" means-O-alkylaryl, wherein alkylaryl is as defined herein.
"substituted arylalkoxy" means-O-alkylaryl, wherein alkylarylSubstantially as defined herein; c in which any aryl radical present may itself be unsubstituted1-C4Alkyl, halogen, cyano, unsubstituted C1-C4Alkoxy, unsubstituted C1-4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"azido" means the group-N3。
"carbamoyl or amido" means the group-C (O) NH2。
"substituted carbamoyl or substituted amido" means the group-C (O) N (R)62)2Wherein each R62Independently are:
·H、C1-C8alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-10 membered heteroaryl and heteroaralkyl; or
C substituted by halogen or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-to 10-membered heteroaryl or heteroaralkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy substitution;
provided that at least one R62Is not H.
Typical "substituted carbamoyl" is-C (O) NR64-C1-C8Alkyl, -C (O) NR64-(CH2)t(C6-C10Aryl), -C (O) N64-(CH2)t(5-10 membered heteroaryl), -C (O) NR64-(CH2)t(C3-C10Cycloalkyl) and-C (O) NR64-(CH2)t(4-10 membered heterocycloalkyl) wherein t is an integer of 0 to 4, R64Each independently represents H or C1-C8Alkyl, optionally present aryl, heteroaryl, cycloalkyl or heterocycloalkyl, which radicals may themselves be unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"carboxy" means the group-C (O) OH.
"cycloalkyl" means a cyclic non-aromatic hydrocarbon group having 3 to 10 carbon atoms. Such cycloalkyl groups include, for example, monocyclic structures such as cyclopropyl, cyclobutyl, cyclopentyl and cyclooctyl.
"substituted cycloalkyl" means a cycloalkyl group as defined above substituted with one or more groups as set forth in the definition of "substituted" herein, and particularly means a cycloalkyl group having 1 or more substituents (e.g., 1 to 5 substituents and particularly 1 to 3 substituents, particularly 1 substituent).
"cyano" means the group-CN.
"halo" or "halogen" means fluorine (F), chlorine (Cl), bromine (Br), and iodine (I). Particular halogen groups are fluorine or chlorine.
"hetero" when used to describe a compound or a group present on a compound means that one or more carbon atoms in the compound or group are replaced with a nitrogen, oxygen, or sulfur heteroatom. Hetero may be applied to any of the above-mentioned hydrocarbon groups, such as alkyl groups, e.g., heteroalkyl groups, cycloalkyl groups, e.g., heterocycloalkyl groups, aryl groups, e.g., heteroaryl groups, cycloalkenyl groups, e.g., cycloheteroalkenyl groups, and the like, having from 1 to 5 and particularly from 1 to 3 heteroatoms.
"heteroaryl" means a monocyclic or polycyclic aromatic ring structure containing one or more heteroatoms and 5 to 12 ring members, more typically 5 to 10 ring members. Heteroaryl groups may, for example, be a 5-or 6-membered monocyclic ring or a bicyclic structure formed by fused 5-and 6-membered rings or two fused 6-membered rings, or as another example a bicyclic structure formed by two fused 5-membered rings. Each ring may contain up to 4 heteroatoms typically selected from nitrogen, sulfur and oxygen. Typically, the heteroaryl ring comprises up to 4 heteroatoms, more typically up to 3 heteroatoms, more typically up to 2 heteroatoms, e.g. a single heteroatom. In one embodiment, the heteroaryl ring comprises at least one ring nitrogen atom. The nitrogen atom on the heteroaryl ring may be basic with respect to imidazole or pyridine, or substantially non-basic with respect to indole or pyrrole nitrogens. In general, the number of basic nitrogen atoms present in the heteroaryl group (including any amino substituents of the ring) is less than 5. Examples of 5-membered monocyclic heteroaryl groups include, but are not limited to, pyrrole, furan, thiophene, imidazole, furazan,azole,Oxadiazole, oxadiazole,Triazole, isopropylOxazole, thiazole, isothiazole, pyrazole, triazole and tetrazolyl. Examples of 6-membered monocyclic heteroaryl groups include, but are not limited to, pyridine, pyrazine, pyridazine, pyrimidine, and triazine. Specific examples of bicyclic heteroaryl groups comprising a 5-membered ring fused to another 5-membered ring include, but are not limited to, imidazothiazole and imidazoimidazole. Specific examples of bicyclic heteroaryls comprising a 6-membered ring fused to a 5-membered ring include, but are not limited to, benzofuran, benzothiophene, benzimidazole, benzo Azole, isobenzoAzole, benzisohOxazole, benzothiazole, benzoisothiazole, isobenzofuran, indole, isoindole, isoindolone, indolizine, indoline, isoindoline, purine (e.g., adenine, guanine), indazole, pyrazolopyrimidine, triazolopyrimidine, benzodioxole, and pyrazolopyridine groups. Specific examples of bicyclic heteroaryls comprising 2 fused 6-membered rings include, but are not limited to, quinoline, isoquinoline, chroman, thiochroman, chromene, isochromene, chroman, isochroman, benzodiazepineAlkane, quinolizine, benzoOxazine, benzodiazine, pyridopyridine, quinoxaline, quinazoline, cinnoline, phthalazine, naphthyridine and pteridine groups. Specific heteroaryl groups are those derived from thiophene, pyrrole, benzothiophene, benzofuran, indole, quinoline, imidazole, and,Groups of oxazoles and pyrazines.
Representative examples of heteroaryl groups include the following:
wherein each Y is selected from carbonyl, N, NR65O and S; r65Independently of each other is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl and 5-10 membered heteroaryl。
Examples of representative aryl groups having heteroatom-containing substitutions include the following:
wherein each W is selected from C (R)66)2、NR66O and S; each Y is selected from carbonyl, NR66O and S; r66Independently of each other is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl and 5-10 membered heteroaryl.
The term "heterocycloalkyl" as used herein means a 4-to 10-membered stable heterocyclic non-aromatic ring and/or includes a ring fused thereto that includes one or more heteroatoms independently selected from N, O and S. Fused heterocyclic ring systems may include carbocyclic rings and need only include one heterocyclic ring. Examples of heterocycles include, but are not limited to, morpholine, piperidine (e.g., 1-piperidinyl, 2-piperidinyl, 3-piperidinyl, and 4-piperidinyl), pyrrolidine (e.g., 1-pyrrolidinyl, 2-pyrrolidinyl, and 3-pyrrolidinyl), pyrrolidone, pyran (2H-pyran or 4H-pyran), dihydrothiophene, dihydropyran, dihydrofuran, dihydrothiazole, tetrahydrofuran, tetrahydrothiophene, dithiane, dihydrofuran, dihydrothiazole, tetrahydrofuran, and mixtures thereofAlkanes, tetrahydropyrans (e.g. 4-tetrahydropyranyl), imidazolines, imidazolidinones (imidazopyridinones),Oxazolines, thiazolines, 2-pyrazolines, pyrazolidines, piperazines and N-alkylpiperazines, for example N-methylpiperazine. Other examples include thiomorpholine and its S-oxides and S, S-dioxides (in particular thiomorpholine). Other examples include azetidines, piperidones, piperazinones, and N-alkylpiperidines, such as N-methylpiperidine. Specific examples of heterocycloalkyl groups are shown in the following illustrative examples ：
Wherein each W is selected from CR67、C(R67)2、NR67O and S; each Y is selected from NR67O and S; r67Independently of each other is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, 5-10 membered heteroaryl. These heterocycloalkyl rings may be optionally substituted with one or more groups selected from acyl, acylamino, acyloxy, alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl (carbamoyl or acylamino), aminocarbonylamino, aminosulfonyl, sulfonylamino, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, halogen, hydroxy, keto, nitro, mercapto, -S-alkyl, -S-aryl, -S (O) -alkyl, -S (O) -aryl, -S (O)2-alkyl and-S (O)2-an aryl group. Substituent groups include carbonyl or thiocarbonyl groups which provide, for example, lactam and urea derivatives.
"hydroxy" means the group-OH.
"Nitro" means the radical-NO2。
"substituted" means a group in which one or more hydrogen atoms are each independently replaced by the same or different substituents. Typical substituents may be selected from:
halogen, -R68、-O-、＝O、-OR68、-SR68、-S-、＝S、-NR68R69、＝NR68、-CCl3、-CF3、-CN、-OCN、-SCN、-NO、-NO2、＝N2、-N3、-S(O)2O-、-S(O)2OH、-S(O)2R68、-OS(O2)O-、-OS(O)2R68、-P(O)(O-)2、-P(O)(OR68)(O-)、-OP(O)(OR68)(OR69)、-C(O)R68、-C(S)R68、-C(O)OR68、-C(O)NR68R69、-C(O)O-,-C(S)OR68、-NR70C(O)NR68R69、-NR70C(S)NR68R69、-NR71C(NR70)NR68R69and-C (NR)70)NR68R69；
Wherein R is68、R69、R70And R71Each independently is:
hydrogen, C1-C8Alkyl radical, C6-C10Aryl, arylalkyl, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, 5-10 membered heteroaryl, heteroarylalkyl; or
C substituted by halogen or hydroxy1-C8An alkyl group; or
·C6-C10Aryl, 5-10 membered heteroaryl, C6-C10Cycloalkyl or 4-to 10-membered heterocycloalkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
In a particular embodiment, a substituted group is substituted with one or more substituents, particularly 1-3 substituents, particularly 1 substituent.
In another specific embodiment, the substituent or group is selected from the group consisting of halogen, cyano, nitro, trifluoromethyl, trifluoromethoxy, azido, -NR72SO2R73、-SO2NR73R72、-C(O)R73、-C(O)OR73、-OC(O)R73、-NR72C(O)R73、-C(O)NR73R72、-NR73R72、-(CR72R72)mOR72Wherein R is73Each independently selected from H, C1-C8Alkyl, - (CH)2)t(C6-C10Aryl), - (CH)2)t(5-10 membered heteroaryl), - (CH)2)t(C3-C10Cycloalkyl) and- (CH)2)t(4-10 membered heterocycloalkyl) wherein t is an integer from 0 to 4; and is
Any alkyl groups present may themselves be substituted by halogen or hydroxy; and is
C where any aryl, heteroaryl, cycloalkyl or heterocycloalkyl radicals present may themselves be unsubstituted1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy. R' each independently represents H or C1-C6An alkyl group.
"substituted thioalkyl" means the radical-SR74Wherein R is74Selected from:
·C1-C8alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-10 membered heteroaryl and heteroaralkyl; or
C substituted by halogen, substituted or unsubstituted amino or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-to 10-membered heteroaryl or heteroaralkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstitutedC1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
A typical "substituted sulfanyl" group is-S- (C)1-C8Alkyl) and-S- (C)3-C10Cycloalkyl), -S- (CH)2)t(C6-C10Aryl), -S- (CH)2)t(5-10 membered heteroaryl), -S- (CH)2)t(C3-C10Cycloalkyl) and-S- (CH)2)t(4-to 10-membered heterocycloalkyl) in which t is an integer from 0 to 4, and C in which any aryl, heteroaryl, cycloalkyl or heterocycloalkyl present may itself be unsubstituted1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy. The term "substituted thioalkyl" includes the groups "alkylsulfanyl" or "alkylthio", "substituted alkylthio" or "substituted alkylsulfanyl", "cycloalkylsulfanyl" or "cycloalkylthio", "substituted cycloalkylsulfanyl" or "substituted cycloalkylthio", "arylsulfanyl" or "arylthio" and "heteroarylsulfanyl" or "heteroarylthio" as defined below.
"alkylthio" or "alkylsulfanyl" means the group-SR75Wherein R is75Is C1-C8Alkyl or a group as defined herein. Representative examples include, but are not limited to, methylthio, ethylthio, propylthio, and butylthio.
"substituted alkylthio" or "substituted alkylsulfanyl" means the group-SR76Wherein R is76Is C substituted by halogen, substituted or unsubstituted amino or hydroxy1-C8An alkyl group.
"Cycloalkylsulfanyl" or "cycloalkylsulfanyl" means the group-SR77Wherein R is77Is C3-C10Cycloalkyl or a group as defined herein. Representative examples include, but are not limited to, cyclopropylthio, cyclohexylthio and cyclopentylthio.
"substituted cycloalkylthio" or "substituted cycloalkylsulfanyl" means the group-SR78Wherein R is78Is C substituted by halogen, substituted or unsubstituted amino or hydroxy3-C10A cycloalkyl group.
"Arylthio" or "arylsulfanyl" means the group-SR79Wherein R is79Is C as defined herein6-C10And (4) an aryl group.
"Heteroarylthio" or "heteroarylsulfanyl" means the group-SR80Wherein R is80Is a 5-10 membered heteroaryl group as defined herein.
"substituted sulfinyl" means the group-S (O) R81Wherein R is81Selected from:
·C1-C8alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-10 membered heteroaryl and heteroaralkyl; or
C substituted by halogen, substituted or unsubstituted amino or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-to 10-membered heteroaryl or heteroaralkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
Typical "substituted sulfinyl" groups are-S (O) - (C)1-C8Alkyl) and-S (O) - (C)3-C10Cycloalkyl), -S (O) - (CH)2)t(C6-C10Aryl), -S (O) - (CH)2)t(5-10 membered heteroaryl), -S (O) - (CH)2)t(C3-C10Cycloalkyl) and-S- (O) - (CH)2)t(4-10 membered heterocycloalkyl) wherein t is an integer from 0 to 4, and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl present may itself be unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy. The term substituted sulfinyl includes the groups "alkylsulfinyl", "substituted alkylsulfinyl", "cycloalkylsulfinyl", "substituted cycloalkylsulfinyl", "arylsulfinyl" and "heteroarylsulfinyl" as defined herein.
"Alkylsulfinyl" means the group-S (O) R82Wherein R is82Is C as defined herein1-C8An alkyl group. Representative examples include, but are not limited to, methylsulfinyl, ethylsulfinyl, propylsulfinyl, and butylsulfinyl.
"substituted alkylsulfinyl" means the group-S (O) R83Wherein R is83Is C substituted as defined herein by halogen, substituted or unsubstituted amino or hydroxy1-C8An alkyl group.
"Cycloalkylsulfinyl" means the group-S (O) R84Wherein R is84Is C3-C10Cycloalkyl or a group as defined herein. Representative examples include, but are not limited to, cyclopropylsulfinyl, cyclohexylsulfinyl, and cyclopentylsulfinyl. Typical "cycloalkylsulfinyl" is S (O) -C3-C10A cycloalkyl group.
"substituted cycloalkanesAlkylsulfinyl "means the group-S (O) R85Wherein R is85Is C substituted by halogen, substituted or unsubstituted amino or hydroxy3-C10A cycloalkyl group.
"Arylsulfinyl" means the group-S (O) R86Wherein R is86Is C as defined herein6-C10And (4) an aryl group.
"heteroarylsulfinyl" means the group-S (O) R87Wherein R is87Is a 5-10 membered heteroaryl group as defined herein.
"substituted sulfonyl" means the group-S (O)2R88Wherein R is88Selected from:
·C1-C8alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-10 membered heteroaryl and heteroaralkyl; or
C substituted by halogen, substituted or unsubstituted amino or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-to 10-membered heteroaryl or heteroaralkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
Typical "substituted sulfonyl" groups are-S (O)2-(C1-C8Alkyl) and-S (O)2-(C3-C10Cycloalkyl), -S (O)2-(CH2)t(C6-C10Aryl), -S (O)2-(CH2)t(5-10 membered heteroaryl), -S (O)2-(CH2)t(C3-C10Cycloalkyl) and-S (O)2-(CH2)t(4-10 membered heterocycloalkyl) wherein t is an integer from 0 to 4 and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl present may itself be unsubstituted1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy. The term substituted sulfonyl includes the groups alkylsulfonyl, substituted alkylsulfonyl, cycloalkylsulfonyl, substituted cycloalkylsulfonyl, arylsulfonyl and heteroarylsulfonyl.
"alkylsulfonyl" means the group-S (O)2R89Wherein R is89Is C as defined herein1-C8An alkyl group. Representative examples include, but are not limited to, methylsulfonyl, ethylsulfonyl, propylsulfonyl, and butylsulfonyl.
"substituted alkylsulfonyl" means the group-S (O)2R90Wherein R is90Is C substituted as defined herein by halogen, substituted or unsubstituted amino or hydroxy1-C8An alkyl group.
"Cycloalkylsulfonyl" means the group-S (O)2R91Wherein R is91Is C3-C10Cycloalkyl or a group as defined herein. Representative examples include, but are not limited to, cyclopropylsulfonyl, cyclohexylsulfonyl, and cyclopentylsulfonyl.
"substituted cycloalkylsulfonyl" means the group-S (O)2R92Wherein R is92Is C substituted by halogen, substituted or unsubstituted amino or hydroxy3-C10A cycloalkyl group.
"arylsulfonyl" means a radical-S (O)2R93Wherein R is93Is C as defined herein6-C10And (4) an aryl group.
"Heteroarylsulfonyl" means a radical-S (O)2R94Wherein R is94Is a 5-10 membered heteroaryl group as defined herein.
"sulfo" or "sulfonic acid" means a group such as-SO3H。
"substituted sulfo" or "sulfonate" means the group-S (O)2OR95Wherein R is95Selected from:
·C1-C8alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-10 membered heteroaryl and heteroaralkyl; or
C substituted by halogen, substituted or unsubstituted amino or hydroxy1-C8An alkyl group; or
·C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-to 10-membered heteroaryl or heteroaralkyl, each of which is unsubstituted C1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
Typical "substituted sulfo" or "sulfonate" groups are-S (O)2-O-(C1-C8Alkyl) and-S (O)2-O-(C3-C10Cycloalkyl), -S (O)2-O-(CH2)t(C6-C10Aryl), -S (O)2-O-(CH2)t(5-10 membered heteroaryl), -S (O)2-O-(CH2)t(C3-C10Cycloalkyl) and-S (O)2-O-(CH2)t(4-10 membered heterocycloalkyl) in which t is an integer from 0 to 4, and any aryl, heteroaryl, cycloalkyl or heterocycloalkyl present may themselves be unsubstitutedC1-C4Alkyl, halogen, unsubstituted C1-C4Alkoxy, unsubstituted C1-C4Haloalkyl, unsubstituted C1-C4Hydroxyalkyl or unsubstituted C1-C4Haloalkoxy or hydroxy.
"mercapto" means the group-SH.
"Aminocarbonylamino" means the group-NR96C(O)NR96R96Wherein each R96Independently of each other is hydrogen, C1-C8Alkyl radical, C3-C10Cycloalkyl, 4-10 membered heterocycloalkyl, C6-C10Aryl, aralkyl, 5-10 membered heteroaryl, and heteroaralkyl as defined herein; or wherein two R are96Groups are linked to form an alkylene group when linked to the same N.
"bicyclic aryl" means a monovalent aromatic hydrocarbon radical derived by the removal of one hydrogen atom from a single carbon atom of a parent bicyclic aromatic ring system. Typical bicyclic aryl groups include, but are not limited to, groups derived from indane, naphthalene, tetrahydronaphthalene, and the like. Specifically, the aryl group contains 8 to 11 carbon atoms.
"bicyclic heteroaryl" means a monovalent bicyclic heteroaromatic group derived by the removal of one hydrogen atom from a single carbon atom of the parent bicyclic heteroaromatic ring system. Typical bicyclic heteroaryl groups include, but are not limited to, those derived from benzofuran, benzimidazole, benzidazole, benzodiazepineAlkane, chromene, chroman, cinnoline, phthalazine, indole, indoline, indolizine, isobenzofuran, isochromene, isoindole, isoindoline, isoquinoline, benzothiazoleAzoles, naphthyridines, benzolsOxadiazole butterflyPyridine, purine, benzopyran, benzopyrazine, pyridopyrimidine, quinazoline, quinoline, quinolizine, quinoxaline, benzomorphane, tetrahydroisoquinoline, tetrahydroquinoline, and the like. Preferred bicyclic heteroaryls are 9-11 membered bicyclic heteroaryls, with 5-10 membered heteroaryls being particularly preferred. Specific bicyclic heteroaryls are those derived from benzothiophene, benzofuran, benzothiazole, indole, quinoline, isoquinoline, benzimidazole, benzo Azole and benzodiThe radical of an alkane.
"Compounds of the invention" and equivalents are intended to include compounds as described above, particularly compounds of any formula described and/or depicted herein, including prodrugs, pharmaceutically acceptable salts, and solvates, e.g., hydrates, where the context allows. Similarly, references to intermediates, whether or not they are themselves claimed, are meant to include salts and solvates thereof, if the context permits.
"cycloalkylalkyl" means a group in which a hydrogen atom of an alkyl group is replaced with a cycloalkyl group. Typical cycloalkylalkyl groups include, but are not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, cycloheptylmethyl, cyclooctylmethyl, cyclopropylethyl, cyclobutylethyl, cyclopentylethyl, cyclohexylethyl, cycloheptylethyl, and cyclooctylethyl, and the like.
"Heterocycloalkylalkyl" means a group in which a hydrogen atom of an alkyl group is replaced with a heterocycloalkyl group. Typical heterocycloalkylalkyl groups include, but are not limited to, pyrrolidinylmethyl, piperidinylmethyl, piperazinylmethyl, morpholinylmethyl, pyrrolidinylethyl, piperidinylethyl, piperazinylethyl, morpholinylethyl, and the like.
"cycloalkenyl" means a cyclic hydrocarbon group having from 3 to 10 carbon atoms and having a single ring or multiple fused rings (including fused and bridged ring systems), and having at least one, and in particular from 1 to 2, sites of ethylenic unsaturation. Such cycloalkenyl groups include, for example, monocyclic structures such as cyclohexenyl, cyclopentenyl, cyclopropenyl, and the like.
"substituted cycloalkenyl" means those groups recited in the definition of "substituted" herein, and specifically means cycloalkenyl having 1 or more (e.g., 1-5 substituents and specifically 1-3 substituents) substituents selected from acyl, amido, acyloxy, alkoxy, substituted alkoxy, alkoxycarbonyl, alkoxycarbonylamino, amino, substituted amino, aminocarbonyl, aminocarbonylamino, aminocarbonyloxy, aryl, aryloxy, azido, carboxyl, cyano, cycloalkyl, substituted cycloalkyl, halogen, hydroxy, keto, nitro, thioalkoxy, substituted thioalkoxy, thioaryloxy, thioketo, mercapto, alkyl-S (O) -, aryl-S (O) -, alkyl-S (O)2And aryl-S (O)2-。
"fused cycloalkenyl" means a cycloalkenyl group having two ring carbon atoms in common with a second aliphatic or aromatic ring and having ethylenic unsaturation located in and imparting aromaticity to the cycloalkenyl group.
"vinyl" means substituted or unsubstituted- (C ═ C) -.
"ethylene" means substituted or unsubstituted- (C-C) -.
"ethynyl" means- (C.ident.C) -.
By "hydrogen bond donor" group is meant a group that contains O-H or N-H functionality. Examples of "hydrogen bond donor" groups include-OH, -NH2and-NH-R97Wherein R is97Is alkyl, acyl, cycloalkyl, aryl or heteroaryl.
"dihydroxyphosphoryl" means the group-PO (OH)2。
"substituted dihydroxyphosphoryl" means those groups described in the definition of "substituted" herein, and specifically means dihydroxyphosphoryl in which one or both hydroxyl groups are substituted. Suitable substituents are described in detail below.
"Aminohydroxyphosphoryl" means the group-PO (OH) NH2。
"substituted aminohydroxyphosphoryl" means those groups described in the definition of "substituted" herein, and in particular aminohydroxyphosphoryl in which an amino group is substituted by one or two substituents. Suitable substituents are described in detail below. In some embodiments, the hydroxyl group may also be substituted.
"nitrogen-containing heterocycloalkyl" means a 4-to 7-membered non-aromatic cyclic group containing at least one nitrogen atom, such as, but not limited to, morpholine, piperidine (e.g., 2-piperidinyl, 3-piperidinyl, and 4-piperidinyl), pyrrolidine (e.g., 2-pyrrolidinyl and 3-pyrrolidinyl), azetidine, pyrrolidone, imidazoline, imidazolidinone, 2-pyrazoline, pyrazolidine, piperazine, and N-alkylpiperazines, such as N-methylpiperazine. Specific examples include azetidines, piperidones, and piperazinones.
"thioketo" means a group that is S.
One of ordinary skill in the art of organic synthesis will appreciate that the maximum number of heteroatoms in a stable, chemically feasible heterocycle, whether aromatic or non-aromatic, is determined by ring size, unsaturation, and heteroatom valency. In general, a heterocycle may have 1-4 heteroatoms, provided that the heterocycle is chemically feasible and stable.
"pharmaceutically acceptable" means approved by a regulatory agency of the federal or a state government or a non-U.S. corresponding regulatory agency or can be approved by it or listed in the U.S. pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans.
By "pharmaceutically acceptable salt" is meant a salt of a compound of the invention that is pharmaceutically acceptable and has the desired pharmacological activity of the parent compound. In particular, such salts are non-toxic and may be inorganic or organic acid addition salts and base addition salts. In particular, such salts include: (1) acid addition salts with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids such as acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3- (4-hydroxybenzoyl) benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1, 2-ethane-disulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo [2.2.2] -oct-2-ene-1-carboxylic acid, glucoheptonic acid, 3-phenylpropionic acid, trimethylacetic acid, tert-butylacetic acid, lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoic acid, salicylic acid, stearic acid, muconic acid, etc.; or (2) salts formed when an acidic proton present on the parent compound is replaced by a metal ion, such as an alkali metal ion, alkaline earth metal ion, or aluminum ion; or a complex with an organic base (e.g., ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, etc.). Salts also include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and when the compound contains a basic functionality, non-toxic salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, and the like. The term "pharmaceutically acceptable cation" means a cationic counterion acceptable for acidic functional groups. Such cations are typically sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium cations, and the like.
By "pharmaceutically acceptable vehicle" is meant a diluent, adjuvant, excipient, or carrier with which a compound of the invention is administered.
By "prodrug" is meant a compound, including derivatives of the compounds of the invention, which have a cleavable group and become the compounds of the invention which are pharmaceutically active in vivo by solvolysis or under physiological conditions. Examples include, but are not limited to, choline ester derivatives and the like, N-alkyl morpholine esters, and the like.
By "solvate" is meant a form of the compound that is typically combined with a solvent by a solvolysis reaction. Such physical bonding includes hydrogen bonding. Common solvents include water, ethanol, acetic acid, and the like. The compounds of the invention may be prepared, for example, in crystalline form, and may be solvated or hydrated. Suitable solvates include pharmaceutically acceptable solvates, such as hydrates, and also includes stoichiometric and non-stoichiometric solvates. In some instances, solvates can separate, for example, when one or more solvent molecules are incorporated into the crystalline solid lattice. "solvate" includes both solution-phase and isolatable solvates. Representative solvates include hydrates, ethanolates, and methanolates.
"subject" includes humans. The terms "human", "patient" and "subject" are used interchangeably herein.
By "therapeutically effective amount" is meant an amount of a compound that, when administered to a subject to treat a disease, is sufficient to treat such disease. The "therapeutically effective amount" may vary depending on the compound, the disease of the subject being treated and its severity and age, weight, etc.
By "preventing" or "prevention" is meant reducing the risk of acquiring or developing a disease or disorder (i.e., causing a subject who has not been exposed to a pathogen or who is predisposed to a disease prior to the onset of the disease to not develop at least one clinical symptom of the disease.
The term "prevention" relates to "prevention" and means a prophylactic measure or method that does not treat or cure a disease. Non-limiting examples of prophylactic measures can include administration of a vaccine; administering low molecular weight heparin to hospitalized patients at risk of thrombosis due to, for example, immobilization; and administering an antimalarial drug, such as chloroquine, prior to visiting a geographic area where malaria is endemic or is at high risk of contracting malaria.
In one embodiment, "treating" or "treatment" of any disease or disorder means ameliorating the disease or disorder (i.e., arresting the disease or reducing the manifestation, extent, or severity of at least one clinical symptom thereof). In another embodiment, "treating" or "treatment" means improving at least one physical parameter (physical parameter), which may not be discernible by the subject. In another embodiment, "treating" or "treatment" means modulating the disease or disorder, either physically (e.g., stabilizing a discernible symptom), physiologically (e.g., stabilizing a physical parameter), or both. In another embodiment, "treating" or "treatment" relates to slowing disease progression.
"Compounds of the invention" and equivalent expressions are meant to include compounds of the formula as described above, including, for example, prodrugs, pharmaceutically acceptable salts and solvates, e.g., hydrates, as the context permits. Similarly, references to intermediates, whether or not they are themselves claimed, are intended to include salts and solvates thereof which are so permitted.
When ranges are referred to herein, for example, but not limited to C1-C8Alkyl, the recitation of a range should be taken as a representation of each member of the range.
Other derivatives of the compounds of the invention are active in their acid and acid derivative forms, but generally offer the advantages of solubility, histocompatibility or delayed release in mammalian organisms in acid sensitive forms (see Bundgard, h., Design of produgs, pp.7-9,21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known to those skilled in the art, such as esters prepared by reacting the parent acid with a suitable alcohol, or amides prepared by reacting the parent acid compound with a substituted or unsubstituted amine, or anhydrides or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups on the side chains of the compounds of the invention are specific prodrugs. In some cases, it is desirable to prepare diester-type prodrugs, such as (acyloxy) alkyl esters or ((alkoxycarbonyl) oxy) alkyl esters. In particular C of the compounds of the invention1－C8Alkyl radical, C2-C8Alkenyl, aryl, C7-C12Substituted byAryl and C7-C12Aryl alkyl esters.
The term "isotopic variation" as used herein means a compound comprising an unnatural proportion of an isotope at one or more of the atoms making up such compound. For example, an "isotopic variant" of a compound can comprise one or more non-radioactive isotopes, such as deuterium (g), (b), (c), (d), and (d)2H or D), carbon-13 (13C) Nitrogen-15 (15N), and the like. It is to be understood that in compounds that are subject to such isotopic substitution, if present, the following atoms may be altered such that, for example, any hydrogen may be2H/D, any carbon may be13C or any nitrogen may be15N, and the presence and configuration of such atoms can be determined by one skilled in the art. Likewise, the invention may include the preparation of isotopic variations using radioisotopes, for example, wherein the resulting compounds may be used in drug and/or substrate tissue distribution studies. With radioactive isotopes of tritium3H and carbon-14 i.e14C is particularly useful for this purpose because of its ease of incorporation and ease of detection. In addition, positron emitting isotopes (e.g. for example) can be prepared11C、18F、15O and13n) and suitable for use in Positron Emission Tomography (PET) studies to examine substrate receptor occupancy.
All isotopic variations (whether radioactive or non-radioactive) of the compounds provided herein are intended to be encompassed within the scope of the present invention.
It will also be understood that compounds having the same molecular formula but differing in nature or in the order of their atomic bonding or in the arrangement of their atoms in space are referred to as "isomers". Isomers that differ in their arrangement in atomic space are referred to as "stereoisomers".
Stereoisomers that are non-mirror images of each other are referred to as "diastereomers", and those that are non-superimposable mirror images of each other are referred to as "enantiomers". For example, when a compound has an asymmetric center, which is bonded to four different groups, then enantiomeric pairs are possible. Enantiomers can be characterized by the absolute configuration of their asymmetric centers and are described by the R-and S-ordering rules of Cahn and Prelog, or by the rotation of the molecule around the plane of polarization, and are called dextrorotatory or levorotatory (i.e., the (+) or (-) -isomers, respectively). The chiral compounds may exist as individual enantiomers or as mixtures thereof. Mixtures containing equal proportions of enantiomers are referred to as "racemic mixtures".
"tautomer" means a compound that is a tautomeric form of the structure of a particular compound and that is altered by substitution of hydrogen atoms and electrons. Thus, the two structures can be in equilibrium by pi electron and atom (usually H) motion. For example, enols and ketones are tautomers because they interconvert rapidly due to treatment with acid or base. Another example of tautomerism is the acid and nitro forms of phenylnitromethane, which are also formed by treatment with acids or bases.
The tautomeric forms may be associated with optimal chemical reactivity and biological activity to yield the compound of interest.
Pure enantiomeric compounds, as used herein, are substantially free of other enantiomers or stereoisomers of the compound (i.e., enantiomeric excess). In other words, the "S" form of the compound is substantially free of the "R" form of the compound and is thus an enantiomeric excess of the "R" form. The term "enantiomerically pure" or "pure enantiomer" means that the compound comprises more than 75%, more than 80%, more than 85%, more than 90%, more than 91%, more than 92%, more than 93%, more than 94%, more than 95%, more than 96%, more than 97%, more than 98%, more than 98.5%, more than 99%, more than 99.2%, more than 99.5%, more than 99.6%, more than 99.7%, more than 99.8%, or more than 99.9% by weight of the enantiomer. In some embodiments, the weight is based on the total weight of all enantiomers or stereoisomers of the compound.
As used herein and unless otherwise indicated, the term "enantiomerically pure R-compounds" means at least about 80% by weight of R-compounds and up to about 20% by weight of S-compounds, at least about 90% by weight of R-compounds and up to about 10% by weight of S-compounds, at least about 95% by weight of R-compounds and up to about 5% by weight of S-compounds, at least about 99% by weight of R-compounds and up to about 1% by weight of S-compounds, at least about 99.9% by weight of R-compounds, or up to about 0.1% by weight of S-compounds. In some embodiments, the weight is based on the total weight of the compound.
As used herein and unless otherwise indicated, the term "enantiomerically pure S-compound" or "S-compound" means at least about 80% by weight of the S-compound and up to about 20% by weight of the R-compound, at least about 90% by weight of the S-compound and up to about 10% by weight of the R-compound, at least about 95% by weight of the S-compound and up to about 5% by weight of the R-compound, at least about 99% by weight of the S-compound and up to about 1% by weight of the R-compound, or at least about 99.9% by weight of the S-compound and up to about 0.1% by weight of the R-compound. In some embodiments, the weight is based on the total weight of the compound.
In the compositions provided herein, an enantiomerically pure compound, or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof, may be present with other active or inactive ingredients. For example, a pharmaceutical composition comprising an enantiomerically pure R-compound may comprise, for example, about 90% excipient and about 10% enantiomerically pure R-compound. In some embodiments, the enantiomerically pure R-compounds in such compositions may, for example, comprise at least about 95% by weight of the R-compound and up to about 5% by weight of the S-compound, based on the total weight of the compound. For example, a pharmaceutical composition comprising an enantiomerically pure S-compound may, for example, comprise about 90% excipient and about 10% enantiomerically pure S-compound. In some embodiments, the enantiomerically pure S-compounds in such compositions may, for example, comprise at least about 95% by weight of the S-compound and up to about 5% by weight of the R-compound, based on the total weight of the compound. In some embodiments, the active ingredient may be formulated with little or no excipient or carrier.
The compounds of the invention may have one or more asymmetric centers; such compounds may thus be prepared as individual (R) -or (S) -stereoisomers or as mixtures thereof.
Unless otherwise indicated, the description or naming of a particular compound in the specification and claims includes the individual enantiomers and mixtures thereof, otherwise racemates thereof. Methods for determining the stoichiometry of stereoisomers and separating stereoisomers are well known in the art.
Compound (I)
In some aspects, provided herein are compounds for use in the prevention and/or treatment of a variety of diseases in a mammal, among which are arthritis, parkinson's disease, alzheimer's disease, stroke, uveitis, asthma, myocardial infarction, the treatment and prevention of pain syndromes (acute and chronic or neuropathic), traumatic brain injury, acute spinal cord injury, neurodegenerative diseases, alopecia (hair loss), inflammatory bowel disease, and autoimmune diseases or disorders.
In one aspect, provided herein are compounds of formula 1:
wherein
B and W are each independently selected from CR4；
X' is selected from CR4aAnd N;
l is-C (R)2aR2b)-；
R1Selected from substituted or unsubstituted aryl, substituted or unsubstituted heteroaryl, hydroxy C1-C4Alkyl radical, C3-C7cycloalkyl-C1-C4Alkyl or 4-7 membered heterocycloalkyl-C1-C4An alkyl group;
R2aand R2bEach independently selected from hydrogen and C1-C4Alkyl or hydroxy C1-C4An alkyl group;
R3is substituted or unsubstituted C1-C6An alkyl group; CH (OH) R3a，OR3a，CN，COR3a，COOR3a，SOR3a，SO2R3a，CONR3aR3b，SONR3aR3bOr SO2NR3aR3b；
R3aIs H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted aryl, or substituted or unsubstituted heteroaryl;
R3bis H, substituted or unsubstituted C1-C6An alkyl group; or R3aAnd R3bTogether form a cycloheteroalkyl ring of 3-7 atoms;
R4each independently selected from the group consisting of H, substituted or unsubstituted alkyl, substituted or unsubstituted acyl, substituted or unsubstituted acylamino, substituted or unsubstituted alkylamino, substituted or unsubstituted alkylthio, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted alkylarylamino, substituted or unsubstituted amino, substituted or unsubstituted arylalkyl, sulfo, substituted sulfonyl, substituted sulfinyl, substituted sulfanyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted arylsulfonyl, azido, substituted or unsubstituted carbamoyl, carboxy, cyano, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocycloalkyl, substituted or unsubstituted dialkylamino, halogen, nitro and mercapto;
R4aAnd R4bEach independently selected from H, substituted or unsubstituted alkyl, substituted or unsubstituted acylSubstituted or unsubstituted acylamino, substituted or unsubstituted alkylamino, substituted or unsubstituted alkylthio, substituted or unsubstituted alkoxy, substituted or unsubstituted alkoxycarbonyl, substituted or unsubstituted alkylarylamino, substituted or unsubstituted arylalkoxy, substituted or unsubstituted amino, substituted or unsubstituted aryl, substituted or unsubstituted arylalkyl, sulfo, substituted sulfonyl, substituted sulfinyl, substituted sulfanyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted arylsulfonyl, azido, substituted or unsubstituted carbamoyl, carboxy, cyano, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloheteroalkyl, substituted or unsubstituted dialkylamino, halogen, heteroaryloxy, substituted or unsubstituted heteroaryl, Substituted or unsubstituted heteroalkyl, hydroxy, nitro and mercapto;
and subscript m' is selected from 0 to 4;
provided that
When R is3Is CO2Me、SO2Ph OR OR3aWhen R is1Is not unsubstituted phenyl;
When R is3Is SO2- (4-methylpiperazin-1-yl) or SO2- (thiomorpholin-1-yl), X' is CH, R4bIs not H;
when R is3When Me is methyl substituted by alkoxy, R1Is not a substituted phenyl group;
when R is3Is CO2When H, R4bIs Cl, F, Br, Me, Et, OMe or CF3；
When X' is CR4a，R3Is CONR3aR3b，R3aWhen is H, R3bIs not substituted n-pentyl, substituted pentynyl, substituted benzyl, substituted phenethyl, substituted thienylethyl or substituted thiazolylethyl; and
when R is1Is a 5-6 membered heterocycloalkylmethyl group, R3Is CO2Me or n-Pr, R4bIs not Cl or 4-F;
or a pharmaceutically acceptable salt, N-oxide, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In another aspect, provided herein is a pharmaceutical composition of a compound of formula 1:
wherein A, B, W, X', L, R1、R3、R4bAnd m' is as described above;
provided that
i) When R is3Is CO2Me、SO2Ph OR OR3aWhen R is1Is not unsubstituted phenyl;
ii) when R is3Is SO2- (4-methylpiperazin-1-yl) or SO2- (thiomorpholin-1-yl), X' is CH, R4bIs not H;
iii) when R is3When Me is methyl substituted by alkoxy, R1Is not a substituted phenyl group;
iv) when R is3Is CO2When H, R4bIs Cl, F, Br, Me, Et, OMe or CF3；
v) when X' is CR4a，R3Is CONR3aR3b，R3aWhen is H, R3bIs not substituted n-pentyl, substituted pentynyl, substituted benzyl, substituted phenethyl, substituted thienylethyl or substituted thiazolylethyl; and
vi) when R is1Is a 5-6 membered heterocycloalkylmethyl group, R3Is CO2Me orn-Pr of R4bIs not Cl or 4-F;
or a pharmaceutically acceptable salt, N-oxide, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
Provided herein in another aspect are pharmaceutical compositions of compounds of formula 1; wherein A, B, W, X', L, R1、R3a、R4bAnd m' is as described above; r3Is CO2Me、SO2Ph OR OR3a；R1Is unsubstituted phenyl.
Provided herein in another aspect are pharmaceutical compositions of compounds of formula 1; a, B, W, L, R therein1、R3aAnd m' is as described above; r3Is SO2- (4-methylpiperazin-1-yl) or SO2- (thiomorpholin-1-yl); x' is CH; r4bIs H.
Provided herein in another aspect are pharmaceutical compositions of compounds of formula 1; wherein A, B, W, X', L, R3a、R4bAnd m' is as described above; r3Is Me or methyl substituted by alkoxy; r1Is a substituted phenyl group.
Provided herein in another aspect are pharmaceutical compositions of compounds of formula 1; wherein A, B, W, X', L, R1、R3aAnd m' is as described above; r3Is CO2H；R4bNot Cl, F, Br, Me, Et, OMe or CF3。
Provided herein in another aspect are pharmaceutical compositions of compounds of formula 1; a, B, W, L, R therein1、R3、R3a、R4bAnd m' is as described above; x' is CR4a；R3Is CONR3aR3b；R3aIs H; and R is3bIs substituted n-pentyl, substituted pentynyl, substituted benzyl, substituted phenethyl, substituted thienylethyl or substituted thiazolylethyl;
herein anotherIn one aspect, there is provided a pharmaceutical composition of a compound of formula 1; wherein A, B, W, X', L, R3aAnd m' is as described above; r1Is 5-6 membered heterocycloalkylmethyl, R3Is CO2Me or n-Pr and R4bIs Cl or 4-F.
In a particular embodiment of formula 1, A, B and W are each CH.
In one embodiment of formula 1, L is selected from-CH2-、-CHMe-、-CMe2-、-CH(CH2OH) -and-CH (CH)2CH2OH)-。
In one embodiment of formula 1, L is selected from-CH2-and-CHMe-.
In one embodiment of formula 1, the compound is a compound of formula 2a, 2b, 2c, 2d, 2e, 2f, 2g, 2h, 2i, 2j, 2k, 2l, 2m, or 2 n:
wherein
X’、R1、R3a、R3b、R4a、R4bAnd m' is as described for formula 1; r2aIs H, Me, CH2OH or CH2CH2OH; het is a substituted or unsubstituted heterocycloalkyl; or a pharmaceutically acceptable salt, solvate, N-oxide, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In one embodiment of formulas 1-2n, subscript m' is 1, 2, or 3.
In one particular embodiment of formulae 1-2n, subscript m' is 1.
In one embodiment of formulas 1-2n, R4bEach independently is H, C1-C4Alkyl, halo C1-C4Alkyl, CN, NO2Or a halogen.
In one embodiment of formula 1, the compound is a compound of formula 3a, 3b, 3c, 3d, 3e, 3f, 3g, 3h, 3i, 3j, 3k, 3l, or 3 m:
wherein
X’、R1、R3a、R3b、R4a、R4bAnd m' is as described for formula 1; r2aIs H, Me, CH2OH or CH2CH2OH; het is a substituted or unsubstituted heterocycloalkyl; or a pharmaceutically acceptable salt, solvate, N-oxide, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In one embodiment of formula 1, the compound is a compound of formula 2b, 2c, 2h, 2i, 3b, 3c, 3h, or 3 i; r3aIs H.
In one embodiment of formulas 1-3n, R3aIs a substituted or unsubstituted alkyl group.
In one embodiment of formulas 1-3n, R3aIs Me, Et, i-Pr, n-Pr, i-Bu, t-Bu, CF3、CH2CF3Or a benzyl group.
In one embodiment of formulas 1-3n, R3aIs a substituted methyl group.
In one embodiment of formulas 1-3n, R3aIs methoxymethyl, methoxyethyl, dimethylaminomethyl or dimethylaminoethyl.
In one embodiment of formulas 1-3n, R3aIs heteroarylmethyl or heterocycloalkylmethyl.
In one of the formulae 1 to 3nIn a particular embodiment, R3aIs heteroarylethyl or heterocycloalkylethyl.
In one embodiment of formulas 1-3n, R3aIs a pyridylmethyl group.
In one embodiment of formulas 1-3n, R3aIs piperidinylmethyl, piperazinylmethyl, pyrrolidinylmethyl, or morpholinylmethyl.
In one embodiment of formulas 1-3n, R3aIs pyridylethyl, piperidinylethyl, piperazinylethyl, pyrrolidinylethyl or morpholinylethyl.
In another specific embodiment of formulas 1-3n, the compound has formula 3b, R3aIs piperidinylmethyl, piperazinylmethyl, pyrrolidinylmethyl, or morpholinylmethyl.
In one embodiment of formulas 1-3n, R3aIs cyclopropyl, cyclopentyl, cyclopropylmethyl or cyclopentylmethyl.
In one embodiment of formulas 1-3n, R3aIs a substituted or unsubstituted heteroaryl.
In one embodiment of formulas 1-3n, R3aIs a substituted or unsubstituted pyridyl, pyrazinyl or pyrimidinyl group.
In one embodiment of formulas 1-3n, R3aIs a substituted or unsubstituted phenyl group.
In one embodiment of formulas 1-3n, R3aIs pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl,Oxazolyl, thiazolyl, triazolyl, thiadiazolyl, unsubstituted or substituted with alkyl or haloalkyl.
In one embodiment of formulas 1-3n, R3aSelected from substituted or unsubstituted quinolinesAryl, isoquinolyl, methylenedioxyphenyl, imidazopyridinyl, benzoAzolyl, benzothiazolyl, and indolyl.
In one embodiment of formulas 1-3n, R3aIs that
And wherein the subscript n1 is selected from 1 to 5, R5aEach independently selected from the group consisting of hydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted acyl, substituted or unsubstituted acylamino, substituted or unsubstituted alkylamino, substituted or unsubstituted alkylthio, substituted or unsubstituted alkoxy, aryloxy, alkoxycarbonyl, substituted or unsubstituted alkylarylamino, arylalkoxy, substituted arylalkoxy, amino, aryl, substituted aryl, arylalkyl, sulfo, substituted sulfinyl, substituted sulfonyl, substituted sulfanyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted arylsulfonyl, azido, substituted or unsubstituted carbamoyl, carboxyl, cyano, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloheteroalkyl, substituted or unsubstituted dialkylamino, substituted or unsubstituted alkylamino, dialkylamino, alkylamino, alkylthio, substituted or unsubstituted alkoxy, aryloxy, alkylthio, substituted or unsubstituted alkoxy, aryloxy, substituted or unsubstituted alkylsulfonyl, Halogen, heteroaryloxy, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroalkyl, hydroxy, nitro and mercapto.
In one embodiment of formulas 1-3n, R3bIs H or alkyl.
In one embodiment of formulas 1-3n, R3bIs H, Me, Et or i-Pr.
In one embodiment of formulas 1-3k, R3bIs H.
In a particular embodiment of formula 2j, 2k, 2j, 3k or 3n, Het is azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, morpholin-1-yl, piperazin-1-yl and azepin-1-yl, unsubstituted or substituted with one or more groups selected from alkyl, alkoxy, dialkylamino, halo, haloalkyl, hydroxy or hydroxyalkyl.
In a particular embodiment of formula 2j, 2k, 2j, 3k or 3n, Het is azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, morpholin-1-yl, piperazin-1-yl and azepin-1-yl, unsubstituted or substituted with one or more groups selected from Me, Et, i-Pr, OMe, NMe2Cl, F, OH or CF3Is substituted with a group (b).
In one embodiment of formula 2l or 3l, R3aIs Me, Et, i-Pr, n-Pr, i-Bu, t-Bu, CF3、CH2CF3Or a benzyl group.
In one embodiment of formula 2l or 3l, R3aIs Me.
In one embodiment of formula 2m or 3m, R3aIs Me, Et or i-Pr.
In one embodiment of formula 2m or 3m, R3bIs Me, Et or i-Pr.
In one embodiment of formula 2m or 3m, R3aAnd R3bEach independently being Me, Et or i-Pr.
In one embodiment of formula 2m or 3m, R3aAnd R3bEach being Me.
In another embodiment of formula 1, the compound is a compound of formula 4, 5, 6, 7, 8, or 9:
wherein
X’、R1、R4bAnd m' is as described for formula 1; r5cIs R5a(ii) a Subscript n3 is 1, 2, or 3; r5aAs described above;
R2aand R2bEach independently selected from hydrogen and C1-C4Alkyl or hydroxy C1-C4An alkyl group; q is-O-or-C (OH) H-;
or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In one embodiment of formulas 4-9, subscript m' is 1, 2, or 3.
In another embodiment of formulas 4-9, subscript m' is 1.
In another embodiment of formula 1, the compound is a compound of formula 10, 11, 12, 13, 14, or 15:
wherein
X’、R1、R4aAnd R4bAs described in formula 1; r5cIs R5a(ii) a Subscript n3 is 1, 2, or 3; r5aAs described above;
R2aand R2bEach independently selected from hydrogen and C1-C4Alkyl or hydroxy C1-C4An alkyl group;
or R2aAnd R2bTogether form a cycloalkyl or cycloheteroalkyl ring of 3-7 atoms; q is-O-or-C (OH) H-;
or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In one embodiment of formulas 4-15, Q is-O-.
In another embodiment of formulas 4-15, Q is-C (OH) H-.
In one embodiment of formulas 4-15, R2aIs H, Me, CH2OH or CH2CH2OH。
In one embodiment of formulas 4-15, R2bIs H.
In one embodiment of formulas 4-15, n3 is 1 or 2.
In one embodiment of formulas 4-15, R5cIndependently selected from the group consisting of H, alkyl, halogen, cyano, alkoxy, and haloalkyl.
In one embodiment of formulas 4-15, R5cIs H, Cl, F, Me, OMe or CF3。
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, X' is CR4a；R4aIs H, C1-C4Alkyl, halo C1-C4Alkyl, CN, NO2Or a halogen.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, X' is CR4a；R4aIs H, Me, CF3Cl, F, CN or NO2。
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, X' is CR4a；R4aIs CN.
In one embodiment of formulas 1, 2a-2N, 3a-3N, and 4-15, X' is N.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R4bIs H, C1-C4Alkyl, halo C1-C4Alkyl or halogen.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15，R4bIs H, Me, CF3Cl, Br or F.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Is a substituted or unsubstituted aryl or heteroaryl group.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Is a substituted or unsubstituted bicyclic aryl, bicyclic alkyl or bicyclic heteroaryl.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15R1Is a substituted or unsubstituted phenyl group.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Is a substituted or unsubstituted pyridyl, pyrazinyl, thiazolyl or pyrimidinyl group.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Is a substituted or unsubstituted pyridyl group.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Is a substituted or unsubstituted pyrimidinyl group.
In a specific embodiment of formulae 1, 2a-2n, 3a-3n, and 4-15, wherein R1Selected from substituted or unsubstituted quinolyl, isoquinolyl, methylenedioxyphenyl, imidazopyridinyl, benzoAzolyl, benzothiazolyl, and indolyl.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Is that
And wherein subscript n2 is selected from 1 to 5 and each R5bIs independently selected fromHydrogen, substituted or unsubstituted alkyl, substituted or unsubstituted acyl, substituted or unsubstituted acylamino, substituted or unsubstituted alkylamino, substituted or unsubstituted alkylthio, substituted or unsubstituted alkoxy, aryloxy, alkoxycarbonyl, substituted or unsubstituted alkylarylamino, arylalkoxy, substituted arylalkoxy, amino, aryl, substituted aryl, arylalkyl, sulfo, substituted sulfinyl, substituted sulfonyl, substituted sulfanyl, substituted or unsubstituted aminosulfonyl, substituted or unsubstituted alkylsulfonyl, substituted or unsubstituted arylsulfonyl, azido, substituted or unsubstituted carbamoyl, carboxyl, cyano, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloheteroalkyl, substituted or unsubstituted dialkylamino, halogen, substituted or unsubstituted alkylamino, alkylthio, substituted or unsubstituted alkoxy, aryloxy, alkylthio, substituted or unsubstituted alkoxy, aryloxy, alkoxycarbonyl, substituted or unsubstituted alkylarylamino, arylthio, substituted or unsubstituted arylalkyl, substituted or unsubstituted carbamoyl, carboxyl, cyano, substituted or unsubstituted cycloalkyl, substituted or unsubstituted cycloheteroalkyl, Heteroaryloxy, substituted or unsubstituted heteroaryl, substituted or unsubstituted heteroalkyl, hydroxy, nitro and mercapto.
In one particular embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, subscript n2 is 1, 2, or 3.
In one particular embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, subscript n2 is 1 or 2.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Is that
And wherein R5bAs described above.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R5bEach independently selected from H, alkyl, halogen, cyano, alkoxy, and haloalkyl.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R5bEach independently selected from H, Me, Et, n-Pr, iso-Pr, Ph, Cl, F, Br, CN,OH、OMe、OEt、OPh、COPh、CO2Me、CH2-N-morpholino, CH2-N- (4-Me-piperidino), NH2、CONH2、CF3、CHF2、OCF3、OCHF2、t-Bu、SMe、CH＝CH-CO2H、SOMe、SO2Me、SO2CF3、SO2NH2,SO3H、SO3Me, cyclopropyl, triazolyl, morpholinyl and pyridyl.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R5bEach independently selected from H, Cl, F, Me, OMe or CF3。
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R5bEach being Me.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R5bEach is OMe.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R5bEach is CF3。
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Selected from hydroxy C1-C4Alkyl radical, C3-C7cycloalkyl-C1-C4Alkyl or 4-7 membered heterocycloalkyl-C1-C4An alkyl group.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Selected from the group consisting of hydroxymethyl, 1-hydroxyethyl and 2-hydroxyethyl.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R1Selected from piperidin-1-ylmethyl, piperazin-1-ylmethyl, and morpholin-1-ylmethyl.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R2aIs hydrogen.
In one embodiment of formulas 1, 2a-2n, 3a-3n and 4-15In the scheme, R2aIs methyl, hydroxymethyl or hydroxyethyl.
In one embodiment of formulas 1, 2a-2n, 3a-3n, and 4-15, R2aIs methyl.
In one embodiment of formula 1, the compound is a compound of formula 3 g; x' is CH.
In one embodiment of formula 1, the compound is a compound of formula 3 g; x' is C-CN.
In one embodiment of formula 1, the compound is a compound of formula 3 g; x' is N.
In one embodiment of formula 1, the compound is a compound of formula 3 g; r2aIs methyl, hydroxymethyl or hydroxyethyl.
In one embodiment of formula 1, the compound is a compound of formula 3 g; r4bIs methyl.
In one embodiment of formula 1, the compound is a compound of formula 3 g; r1Is that
And wherein R5bIs Me, CF3Or OMe.
In one embodiment of formula 1, the compound is a compound of formula 3 g; r1Is that
And wherein R5bIs Me, CF3Or OMe.
In one embodiment of formula 1, are combinedCompound of formula 3 g; r1Is that
And wherein R5bIs Me, CF3Or OMe.
In one embodiment of formula 1, the compound is a compound of formula 3 g; x' and R1、R2aAnd R4bAs described in the preceding paragraph, R3aIs Me or Et.
In one embodiment of formula 1, the compound is a compound of formula 3 g; x' and R1、R2aAnd R4bAs described in the preceding paragraph, R3aIs Me.
In another embodiment of formula 1, the compound is a compound of formula 16, 17, 18, 19, 20, or 21:
wherein
R2aIs H, Me, CH2OH or CH2CH2OH；R4aIs H or CN; r4bIs F, Cl, Br, Me or CF3；R5bIs F, Cl, Br, Me, OMe or CF3；
R3Is i) CH2R3c；ii)C(OH)(H)R3a；iii)C(OH)(Me)R3a；iv)OR3a；v)C(＝O)R3a；vi)C(＝O)OR3a；vii)S(O)2R3a；viii)C(＝O)NR3aR3b；ix)S(O)2NR3aR3b；x)C(＝O)-Het；xi)S(O)2-Het；xii)CH2OR3a；xiii)CH2NR3aR3b(ii) a And xiv) C (OH) (H) CH2-Het；
And R is3a、R3b、R3cAnd Het is as described for formulae 2a-3 n;
or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In one embodiment of formulas 16-21, R5bIs Me.
In one embodiment of formulas 16-21, R5bIs CF3。
In one embodiment of formulas 16-21, R5bIs OMe.
In another embodiment of formula 1, the compound is a compound of formula 22 or 23:
Wherein
Cy is a substituted or unsubstituted 4-7 membered N-containing heterocycloalkyl group;
R2ais H, Me, CH2OH or CH2CH2OH；R4aIs H or CN; r4bIs F, Cl, Br, Me or CF3；
R3Is i) CH2R3c；ii)C(OH)(H)R3a；iii)C(OH)(Me)R3a；iv)OR3a；v)C(＝O)R3a；vi)C(＝O)OR3a；vii)S(O)2R3a；viii)C(＝O)NR3aR3b；ix)S(O)2NR3aR3b；x)C(＝O)-Het；xi)S(O)2-Het；xii)CH2OR3a；xiii)CH2NR3aR3b(ii) a Or xiv) C (OH) (H) CH2-Het；
And R is3a、R3b、R3cAnd Het is as described for formulae 2a-3 n;
or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In one embodiment of formulas 22-23, Cy is a morpholin-1-yl.
In one embodiment of formulas 16-23, R2aIs H.
In one embodiment of formulas 16-23, R2aIs Me.
In one embodiment of formulas 16-23, R2aIs CH2OH。
In one embodiment of formulae 16-18 and 22, R4aIs H.
In one embodiment of formulae 16-18 and 22, R4aIs CN.
In one embodiment of formulas 16-23, R4bIs Me.
In one embodiment of formulas 16-23, R3aIs Me, Et, i-Pr, n-Pr, i-Bu, t-Bu, CF3、CH2CF3Or a benzyl group.
In one embodiment of formulas 16-23, R3aIs methoxymethyl, methoxyethyl, dimethylaminomethyl or dimethylaminoethyl.
In one embodiment of formulas 16-23, R3aIs piperidinylmethyl, piperazinylmethyl, pyrrolidinylmethyl, or morpholinylmethyl.
In one embodiment of formulas 16-23, R3aIs pyridylmethyl, pyridylethyl, piperidinylethyl, piperazinylethyl, pyrrolidinylethyl or morpholinylethyl.
An implement in the formulae 16-23In a particular embodiment, R3aIs cyclopropyl, cyclopentyl, cyclopropylmethyl or cyclopentylmethyl.
In one embodiment of formulas 16-23, R3aIs a substituted or unsubstituted phenyl, pyridyl, pyrazinyl or pyrimidinyl group.
In one embodiment of formulas 16-23, R3aIs pyrrolyl, furyl, thienyl, pyrazolyl, imidazolyl,Oxazolyl, thiazolyl, triazolyl, thiadiazolyl, unsubstituted or substituted with alkyl or haloalkyl.
In one embodiment of formulas 16-23, R3aIs quinolinyl, isoquinolinyl, methylenedioxyphenyl, imidazopyridinyl, benzoAzolyl, benzothiazolyl, and indolyl.
In one embodiment of formulas 16-23, R3bIs H, Me, Et or i-Pr.
In one embodiment of formulas 16-23, R3bIs H.
In one particular embodiment of formulae 16-23, Het is azetidin-1-yl, pyrrolidin-1-yl, piperidin-1-yl, morpholin-1-yl, piperazin-1-yl, and azepin-1-yl, which is unsubstituted or substituted with one or more groups selected from Me, Et, i-Pr, OMe, NMe2Cl, F, OH or CF3Is substituted with a group (b).
In one embodiment of formula 1, the compound is selected from the compounds listed in table 1.
In one embodiment of formula 1, the compound is selected from
4' -methyl-5- (propane-1-sulfonyl) -biphenyl-3-carboxylic acid ((S) -2-hydroxy-1-methyl-ethyl) -amide;
2,4' -dimethyl-5- (pyrrolidine-1-sulfonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
2 '-cyano-4' -methyl-5- (pyrrolidine-1-sulfonyl) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (pyrrolidine-1-sulfonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (2-methyl-propane-1-sulfonyl) -biphenyl-3-carboxylic acid ((S) -2-hydroxy-1-methyl-ethyl) -amide;
5-cyclopentanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid ((S) -2-hydroxy-1-methyl-ethyl) -amide;
4' -methyl-5- (propane-2-sulfonyl) -biphenyl-3-carboxylic acid ((S) -2-hydroxy-1-methyl-ethyl) -amide;
3-methanesulfonyl-5- (5-methyl-pyridin-2-yl) -N- (2-methyl-pyrimidin-5-ylmethyl) -benzamide;
3- (4-hydroxy-pyrrolidin-3-yloxy) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -benzamide;
5- (2-dimethylamino-1-hydroxy-ethyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (4-methyl-3-oxo-piperazine-1-carbonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (2-methyl-aziridine-1-carbonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid ((S) -1-methyl-2-morpholin-4-yl-ethyl) -amide;
5- [ 1-hydroxy-2- (4-methyl-piperazin-1-yl) -ethyl ] -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide; and
5- { 1-hydroxy-2- [ (2-hydroxy-ethyl) -methyl-amino ] -ethyl } -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
In one embodiment of formula 1, the compound is selected from
4' -methyl-biphenyl-3, 5-dicarboxylic acid 5- [ (6-chloro-pyridin-3-ylmethyl) -amide ]3- (isobutyl-methyl-amide);
4' -methyl-biphenyl-3, 5-dicarboxylic acid 3- (isobutyl-methyl-amide) 5- [ (2-methyl-pyrimidin-5-ylmethyl) -amide ];
4' -methyl-biphenyl-3, 5-dicarboxylic acid 3- (isobutyl-methyl-amide) 5- { [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide };
4' -methyl-5- [ (2-methyl-pyrimidin-5-ylmethyl) -carbamoyl ] -biphenyl-3-carboxylic acid ethyl ester;
5- (3, 3-difluoro-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (piperidine-1-carbonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5- (azepane-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [1- (4-chloro-3-methanesulfonyl-phenyl) -ethyl ] -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5- (azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5- (3-methoxy-pyrrolidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (2-trifluoromethyl-pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5- (3, 3-difluoro-pyrrolidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-trifluoromethyl-pyridin-3-yl) -ethyl ] -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid 3-methanesulfonyl-4-methyl-benzylamide;
2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (propane-1-sulfonyl) -biphenyl-3-carboxylic acid 3-methanesulfonyl-4-methyl-benzylamide;
4' -methyl-5- (propane-1-sulfonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (2-methoxy-pyrimidin-5-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (imidazo [1,2-a ] pyridin-7-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (5-methyl-pyrazin-2-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -methyl-5- (propane-1-sulfonyl) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -methyl-5- (propane-1-sulfonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (propane-1-sulfonyl) -biphenyl-3-carboxylic acid (imidazo [1,2-a ] pyridin-7-ylmethyl) -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
2 '-cyano-5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
2 '-cyano-5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-trifluoromethyl-pyridin-3-ylmethyl) -amide;
2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (6-chloro-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid 3-methanesulfonyl-4-methyl-benzylamide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (1, 1-dioxo-2, 3-dihydro-1H-1 $ l% 6& -benzo [ b ] thiophen-6-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (3-chloro-5-trifluoromethyl-pyridin-2-ylmethyl) -amide;
5- (azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-trifluoromethyl-pyridin-3-yl) -ethyl ] -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5- (azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methoxy-pyrimidin-5-yl) -ethyl ] -amide;
5- (azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (3, 5-dichloro-pyridin-2-ylmethyl) -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-trifluoromethyl-pyridin-3-yl) -ethyl ] -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (S) -1- (6-difluoromethyl-pyridin-3-yl) -2-hydroxy-ethyl ] -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (benzo)Oxazol-5-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (6-chloro-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (S) -2-hydroxy-1- (6-trifluoromethyl-pyridin-3-yl) -ethyl ] -amide;
5- ((R) -3-hydroxy-pyrrolidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (S) -2-hydroxy-1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid 4-chloro-3- [1,2,4] triazol-4-yl-benzylamide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
4' -methyl-5- (pyrrolidine-1-sulfonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
3- (5-methyl-pyridin-2-yl) -N- (6-methyl-pyridin-3-ylmethyl) -5- (pyrrolidine-1-sulfonyl) -benzamide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (R) -1- (5-chloro-1-methyl-1H-pyrazol-4-yl) -ethyl ] -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
3- (5-methyl-pyridin-2-yl) -N- [1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (pyrrolidine-1-carbonyl) -benzamide;
3- (5-methyl-pyridin-2-yl) -N- (6-methyl-pyridin-3-ylmethyl) -5- (pyrrolidine-1-carbonyl) -benzamide;
3- (5-methyl-pyridin-2-yl) -5- (pyrrolidine-1-carbonyl) -N- (6-trifluoromethyl-pyridin-3-ylmethyl) -benzamide;
5- (hexahydro-pyrrolo [1,2-a ] pyrazine-2-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (3-chloro-5-trifluoromethyl-pyridin-2-ylmethyl) -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-trifluoromethyl-pyridin-3-ylmethyl) -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-chloro-pyridin-3-ylmethyl) -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
5- (3-hydroxy-azetidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (3, 5-dichloro-pyridin-2-ylmethyl) -amide;
3- (5-methyl-pyridin-2-yl) -5- (pyrrolidine-1-carbonyl) -N- [ (R) -1- (6-trifluoromethyl-pyridin-3-yl) -ethyl ] -benzamide;
3- ((R) -3-hydroxy-pyrrolidine-1-carbonyl) -5- (5-methyl-pyridin-2-yl) -N- (6-trifluoromethyl-pyridin-3-ylmethyl) -benzamide;
3- ((R) -3-hydroxy-pyrrolidine-1-carbonyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-trifluoromethyl-pyridin-3-yl) -ethyl ] -benzamide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (6-methoxy-pyridin-3-ylmethyl) -amide;
5- ((R) -3-hydroxy-pyrrolidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5- ((R) -3-hydroxy-pyrrolidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-trifluoromethyl-pyridin-3-yl) -ethyl ] -amide;
4' -methyl-biphenyl-3, 5-dicarboxylic acid 3- (methyl-pyridin-4-ylmethyl-amide) 5- [ (6-methyl-pyridin-3-ylmethyl) -amide ];
4' -methyl-biphenyl-3, 5-dicarboxylic acid 5- [ (6-methyl-pyridin-3-ylmethyl) -amide ]3- [ methyl- (2,2, 2-trifluoro-ethyl) -amide ];
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (pyrrolidine-1-carbonyl) -benzamide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -5- (pyrrolidine-1-carbonyl) -benzamide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- ((R) -3-hydroxy-pyrrolidine-1-carbonyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
3- ((R) -3-hydroxy-pyrrolidine-1-carbonyl) -5- (5-methyl-pyridin-2-yl) -N- (6-methyl-pyridin-3-ylmethyl) -benzamide;
4' -methyl-5- (2-methyl-propane-1-sulfonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (2-methyl-propane-1-sulfonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5- (2, 5-dimethyl-pyrrolidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
5- (hexahydro-pyrrolo [1,2-a ] pyrazine-2-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5- (hexahydro-pyrrolo [1,2-a ] pyrazine-2-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (7-aza-bicyclo [2.2.1] heptane-7-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (2-methyl-propane-1-sulfonyl) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5- ((2R,5R) -2, 5-dimethyl-pyrrolidine-1-carbonyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (propane-2-sulfonyl) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
4' -methyl-5- (propane-2-sulfonyl) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -methyl-5- (propane-2-sulfonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -bromo-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (propane-2-sulfonyl) -biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -3-hydroxy-1- (6-methyl-pyridin-3-yl) -propyl ] -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (S) -2-hydroxy-1- (6-methoxy-pyridin-3-yl) -ethyl ] -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (S) -1- (6-difluoromethyl-pyridin-3-yl) -2-hydroxy-ethyl ] -amide;
5-ethanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5-ethanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5-ethanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
5-ethanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
4' -bromo-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -bromo-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5-cyclopentanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5-cyclopentanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (S) -2-hydroxy-1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -3-hydroxy-1- (6-methoxy-pyridin-3-yl) -propyl ] -amide;
4' -methyl-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (6-methyl-1-oxy-pyridin-3-ylmethyl) -amide;
4' -methyl-5- (pyrimidin-2-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
4' -methyl-5- (thiazol-2-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -methyl-5- (thiazol-2-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (hydroxy-pyridin-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
5- (hydroxy-pyridin-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (2-methoxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
5- (2-methoxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (2-methoxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-5- (2-methoxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5- (2-methoxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (thiazol-2-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (thiazol-2-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid (6-methyl-1-oxy-pyridin-3-ylmethyl) -amide;
5-methanesulfonyl-4' -methyl-biphenyl-3-carboxylic acid [ (S) -2-hydroxy-1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (pyridin-2-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (thiazol-2-yloxy) -benzamide;
5- (hydroxy-pyridin-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid (2-methyl-pyrimidin-5-ylmethyl) -amide;
5- (hydroxy-pyridin-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -5- (thiazol-2-yloxy) -benzamide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -5- (pyridin-2-yloxy) -benzamide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (pyridin-2-yloxy) -benzamide;
3- (hydroxy-phenyl-methyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
3- (hydroxy-phenyl-methyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -benzamide;
2 '-cyano-5- (hydroxy-phenyl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-5- (hydroxy-phenyl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (hydroxy-thiazol-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (hydroxy-thiazol-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-5- (hydroxy-pyridin-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5- (hydroxy-pyridin-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
3- (hydroxy-pyridin-2-yl-methyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -benzamide;
3- (hydroxy-pyridin-2-yl-methyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
5- (1, 2-dihydroxy-ethyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5-hydroxymethyl-4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (hydroxy-thiazol-2-yl-methyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
3- (hydroxy-thiazol-2-yl-methyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -benzamide;
2 '-cyano-5- (hydroxy-thiazol-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-5- (hydroxy-thiazol-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (2,2, 2-trifluoro-1-hydroxy-ethyl) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (2,2, 2-trifluoro-1-hydroxy-ethyl) -biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
2 '-cyano-5- (1, 2-dihydroxy-ethyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (2-methoxy-1-methyl-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (2-methoxy-1-methyl-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
5- (2-hydroxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5- (2-hydroxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
4' -methyl-5- (tetrahydro-furan-3-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
4' -methyl-5- (tetrahydro-furan-2-ylmethoxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5- (1-hydroxy-ethyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (tetrahydro-furan-2-ylmethoxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (tetrahydro-furan-3-ylmethoxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (tetrahydro-furan-3-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5- (2-methoxy-1-methyl-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2' -cyano-5- ([1, 4)]IIAlk-2-ylmethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl]-an amide;
5- (2, 3-dihydroxy-propoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (1-hydroxy-ethyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (1-hydroxymethyl-2-methoxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (2-methoxy-1-methyl-ethoxy) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
5- [ hydroxy- (3-methyl-3H-imidazol-4-yl) -methyl ] -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (1-hydroxy-ethyl) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (tetrahydro-furan-3-yloxy) -benzamide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (tetrahydro-furan-3-ylmethoxy) -benzamide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (tetrahydro-furan-2-ylmethoxy) -benzamide;
5- (4-hydroxy-tetrahydro-furan-3-yloxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5- (4-hydroxy-tetrahydro-furan-3-yloxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (2-hydroxy-1-methyl-ethoxy) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
5- (2-hydroxy-1-methyl-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (2-hydroxy-1-methoxymethyl-ethoxy) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
2 '-cyano-4' -methyl-5- (2,2, 2-trifluoro-1-hydroxy-ethyl) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5- [ hydroxy- (3-methyl-3H-imidazol-4-yl) -methyl ] -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-4' -methyl-5- (tetrahydro-furan-3-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
4' -methyl-5- (tetrahydro-furan-3-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5- (4-hydroxy-tetrahydro-furan-3-yloxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
2 '-cyano-5- (4-hydroxy-tetrahydro-furan-3-yloxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (2-methyl-pyrimidin-5-yl) -ethyl ] -amide;
5- (1-hydroxy-2-morpholin-4-yl-ethyl) -4' -methyl-biphenyl-3-carboxylic acid (6-methyl-pyridin-3-ylmethyl) -amide;
3- (5-hydroxymethyl-thiazol-2-yloxy) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
2 '-cyano-4' -methyl-5- (4-methyl-thiazol-2-yloxy) -biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
3- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -5- (4-methyl-thiazol-2-yloxy) -benzamide;
3- (benzothiazol-2-yloxy) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
2 '-cyano-5- (2-hydroxy-1-methyl-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
2 '-cyano-5- (1-hydroxymethyl-2-methoxy-ethoxy) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (1-hydroxy-propyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide;
5- (1, 2-dimethoxy-ethyl) -4' -methyl-biphenyl-3-carboxylic acid [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -amide; and
3- (4-chloro-thiazol-2-yloxy) -5- (5-methyl-pyridin-2-yl) -N- [ (R) -1- (6-methyl-pyridin-3-yl) -ethyl ] -benzamide;
or a pharmaceutically acceptable salt, solvate, prodrug, stereoisomer, tautomer, or isotopic variant thereof.
Other embodiments within the scope provided herein are set forth in non-limiting manner in other sections and examples herein. It is to be understood that these examples are for illustrative purposes only and are not to be construed as limiting in any way.
In some aspects, provided herein are prodrugs and derivatives of the compounds of the above formula. Prodrugs are derivatives of the compounds provided herein that have groups that can be cleaved by metabolic means and converted by solvolysis or under physiological conditions to the compounds provided herein, which are pharmaceutically active in vivo. Examples include, but are not limited to, choline ester derivatives and the like, N-alkyl morpholine ester derivatives, and the like.
Some of the compounds provided herein are active in their acid and acid derivative forms, but generally offer the advantages of solubility, histocompatibility or delayed release in mammalian organisms in acid sensitive forms (see Bundgard, h., Design of produgs, pp.7-9,21-24, Elsevier, Amsterdam 1985). Prodrugs include acid derivatives well known to those skilled in the art, such as esters prepared by reacting the parent acid with a suitable alcohol, or amides prepared by reacting the parent acid compound with a substituted or unsubstituted amine, or anhydrides or mixed anhydrides. Simple aliphatic or aromatic esters, amides and anhydrides derived from acidic groups on the side chains of the compounds of the present invention are preferred prodrugs. In some cases, it is desirable to prepare diester-type prodrugs such as (acyloxy) alkyl esters or ((alkoxycarbonyl) oxy) alkyl esters. Preference is given to C of the compounds provided herein1－C8Alkyl radical, C2-C8Alkenyl, aryl, C7-C12Substituted aryl and C7-C12Aryl alkyl esters.
Pharmaceutical composition
When used as a medicament, the compounds provided herein are typically administered in the form of a pharmaceutical composition. Such compositions may be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound.
In general, the compounds provided herein are administered in a therapeutically effective amount. The amount of compound actually administered is typically determined by a clinician according to relevant circumstances including the disease being treated, the chosen route of administration, the actual compound administered, the age, weight and response of the individual patient, the severity of the patient's symptoms, and the like.
The pharmaceutical compositions provided herein can be administered by a variety of routes, including oral, rectal, transdermal, subcutaneous, intravenous, intramuscular, and intranasal routes. Depending on the intended route of delivery, the compounds provided herein are preferably formulated as injectable or oral compositions or as ointments, lotions or patches, all for transdermal administration.
Compositions for oral administration may take the form of bulk liquid solutions or suspensions or bulk powders. However, it is more common to formulate the compositions in unit dosage form that facilitates accurate administration. The term "unit dosage form" means physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include prefilled, pre-measured ampoules or syringes of liquid compositions, or in the case of solid compositions, pills, tablets, capsules and the like. In such compositions, the furansulfonic acid compound is typically a minor ingredient (about 0.1 to about 50% by weight or preferably about 1 to about 40% by weight), the remainder being various vehicles or carriers and processing aids that aid in forming the desired administration form.
Liquid forms suitable for oral administration may include suitable aqueous or nonaqueous carriers containing buffers, suspending and dispersing agents, coloring agents, flavoring agents, and the like. Solid forms may include, for example, any of the following or compounds of similar nature: binders, such as microcrystalline cellulose, gum tragacanth or gelatin; excipients, such as starch or lactose; disintegrating agents, such as alginic acid, Primogel or corn starch; lubricants, such as magnesium stearate; glidants, such as colloidal silicon dioxide; sweetening agents, such as sucrose or saccharin; or a flavoring agent, such as peppermint, methyl salicylate, or orange flavoring.
Injectable compositions are typically based on injectable sterile saline or phosphate buffered saline or other injectable carriers well known in the art. As noted above, the active compound in such compositions is typically a minor ingredient, usually about 0.05-10% by weight, with the remainder being an injectable carrier or the like.
Transdermal compositions are typically formulated as topical ointments or creams containing the active ingredient, generally in an amount of about 0.01 to about 20% by weight, preferably about 0.1 to about 10% by weight, and more preferably about 0.5 to about 15% by weight. When formulated as an ointment, the active ingredient is typically mixed with either paraffin or with a water-miscible ointment base. Alternatively, the active ingredient may be formulated as a cream, for example, in an oil-in-water cream base. Such transdermal formulations are well known in the art and typically include additional ingredients that promote the stability of the active ingredient or epidermal penetration of the formulation. All such transdermal formulations and ingredients are included within the scope provided herein.
The compounds provided herein can also be administered by transdermal means. Thus, transdermal administration can be carried out using patches of the reservoir or porous membrane type or of various solid matrices.
The above ingredients for orally, parenterally or topically administrable compositions are representative only. Other materials and processing techniques, etcRemington’sPharmaceuticalSciences17 th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, section 8, which is incorporated herein by reference.
The above ingredients for orally, parenterally or topically administrable compositions are representative only. Other materials and processing techniques are set forth in Remington's The Science and Practice of Pharmacy,21st edition,2005, Publisher: Lippincott Williams & Wilkins, section 8, which is incorporated herein by reference.
The compounds of the present invention may also be administered in a sustained release form or from a slow release drug delivery system. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.
The following formulation examples illustrate representative pharmaceutical compositions that can be prepared according to the present invention. However, the present invention is not limited to the following pharmaceutical compositions.
Formulation 1-tablet
The compounds of the invention may be mixed as a dry powder with a dry gelatin binder in a weight ratio of about 1: 2. A small amount of magnesium stearate was added as a lubricant. The mixture was formed into 240-270mg tablets (80-90mg active compound/tablet) using a tablet press.
Preparation 2-capsule
The compounds of the present invention may be mixed with a starch diluent in a weight ratio of about 1:1 as a dry powder. The mixture was filled into 250mg capsules (125mg active compound/capsule).
Formulation 3-liquid
The compound of the invention (125mg) may be mixed with sucrose (1.75g) and xanthan gum (4mg), the resulting mixture may be blended, passed through a No. 10 U.S. sieve, and then mixed with a previously prepared aqueous solution of microcrystalline cellulose and sodium carboxymethylcellulose (11:89,50 mg). Sodium benzoate (10mg), flavours and colours were diluted with water and added while stirring. Sufficient water was then added to produce a total volume of 5 mL.
Formulation 4-tablet
The compounds of the invention may be mixed with dry gelatin binder in a weight ratio of about 1:2 as a dry powder. A small amount of magnesium stearate was added as a lubricant. The mixture was formed into 450-900mg tablets (150-300mg active compound/tablet) using a tablet press.
Preparation 5-injection
The compounds of the present invention may be dissolved or suspended in a buffered sterile saline injectable aqueous medium to a concentration of about 5 mg/mL.
Formulation 6-topical formulation
Stearyl alcohol (250g) and white petrolatum (250g) may be melted at about 75 ℃, then a mixture of the compound of the present invention (50g), methylparaben (0.25g), propylparaben (0.15g), sodium lauryl sulfate (10g) and propylene glycol (120g) dissolved in water (about 370g) is added and the resulting mixture is stirred until it coagulates.
Method of treatment
The compounds of the present invention are useful as therapeutic agents for the treatment of diseases in mammals. Accordingly, the compounds and pharmaceutical compositions provided herein are useful as therapeutic agents for the prevention and/or treatment of neurodegenerative, autoimmune and inflammatory diseases in mammals, including humans and non-human mammals. Accordingly and as indicated above, the present invention includes within its scope and extends to such methods of treatment, to compounds for use in such methods, and to the use of such compounds in the manufacture of medicaments for use in such methods.
In one aspect of the treatment methods, provided herein are methods of treating a mammal susceptible to or suffering from a disease associated with arthritis, asthma, myocardial infarction, inflammatory bowel disease, and autoimmune diseases, comprising administering an effective amount of one or more of the pharmaceutical compositions.
In another aspect of the treatment methods, provided herein are methods of treating a mammal susceptible to or suffering from a disease that produces a painful response or involves the maintenance of an imbalance in sensory nerve basal activity. The compounds of the present invention are useful as analgesics for the treatment of pain of various origins or etiologies, such as acute, inflammatory pain (e.g., pain associated with osteoarthritis and rheumatoid arthritis); various neuropathic pain syndromes (e.g., post-herpetic neuralgia, trigeminal neuralgia, reflex sympathetic dystrophy, diabetic neuropathy, Guillain-Barr syndrome, fibromyalgia, phantom limb pain, post-mastectomy pain, polyneuropathy, HIV neuropathy, and chemotherapy-induced neuropathy and other iatrogenic neuropathies); visceral pain (e.g., visceral pain associated with gastroesophageal reflux disease, irritable bowel syndrome, inflammatory bowel disease, pancreatitis, and various gynecological and urological disorders), dental pain, and headache (e.g., migraine, cluster headache, and tension headache).
In other aspects of the methods of treatment, provided herein are methods of treating a mammal susceptible to or suffering from neurodegenerative diseases and disorders, such as parkinson's disease, alzheimer's disease, and multiple sclerosis; diseases and disorders mediated by or resulting in neuroinflammation, such as encephalitis; centrally mediated neuropsychiatric diseases and disorders such as depression, mania, bipolar disorder, anxiety, schizophrenia, eating disorders, sleep disorders, and cognitive disorders; epilepsy and seizures; prostate, bladder and bowel dysfunction, such as urinary incontinence, urinary hesitancy, rectal hypersensitivity, fecal incontinence, benign prostatic hypertrophy and inflammatory bowel disease; respiratory and airway diseases and disorders such as allergic rhinitis, asthma and reactive airway disease and chronic obstructive pulmonary disease; diseases and disorders mediated by or resulting in inflammation, such as rheumatoid arthritis and osteoarthritis, myocardial infarction, various autoimmune diseases and disorders; itch/pruritus, such as psoriasis; obesity; disorders of lipid metabolism; cancer; and a renal disease, the method comprising administering the one or more pharmaceutical compositions in an amount effective to treat the disease or prevent the disease.
As a further aspect, there are provided compounds of the invention for use as medicaments, particularly in the treatment or prevention of the above-mentioned conditions and diseases. We also provide the use of a compound of the invention in the manufacture of a medicament for the treatment or prevention of one of the conditions and diseases described above.
The injection dosage level is from about 0.1 mg/kg/hr to at least 10 mg/kg/hr, all within about 1 to about 120 hours, especially 24 to 96 hours. A bolus injection of about 0.1mg/kg to about 10mg/kg or more may also be administered to achieve sufficient steady state levels. It is not desirable for 40-80 kg human patients to have a maximum total dose of more than about 2 g/day.
For the prevention and/or treatment of long-term diseases, such as neurodegenerative and autoimmune diseases, the treatment regimen usually lasts for many months or years, and therefore oral administration is preferred for patient compliance and tolerability. Because of the oral administration, 1-5 and especially 2-4 and typically 3 oral doses per day are representative regimens. Using these modes of administration, each dose provides about 0.01 to about 20mg/kg of a compound provided herein, with preferred doses each providing about 0.1 to about 10mg/kg, especially about 1 to about 5 mg/kg.
The transdermal dose is generally selected to provide a blood level similar to or lower than that achieved using an injected dose.
When used to prevent the onset of a neurodegenerative, autoimmune, or inflammatory disease, the compounds provided herein are administered to a patient at risk of developing the disease, typically at the dosage levels described above under the direction and supervision of a clinician. Patients at risk for developing a particular disease generally include patients with a family history or who have been genetically determined or screened to be particularly susceptible to developing the disease.
The compounds provided herein can be administered as a single active agent, or they can be administered in combination with other agents, including other active amines and derivatives. The combined administration may be carried out by any technique apparent to those skilled in the art, including, for example, separate, sequential, simultaneous and alternating administration.
General synthetic methods
The compounds provided herein can be prepared from readily available starting materials using the following general methods and procedures. See, for example, synthetic schemes 1-11 below. It will be understood that typical or preferred processing conditions (i.e., reaction temperatures, times, molar ratios of reactants, solvents, pressures, etc.) are given therein, and other conditions may be used unless otherwise stated. The optimum reaction conditions will vary depending on the particular reactants or solvents used, but such conditions can be determined by one skilled in the art by routine optimization methods.
In addition, as will be apparent to those skilled in the art, conventional protecting groups are required to prevent undesired reactions of the functional groups. The selection of suitable protecting groups for particular functional groups and suitable conditions for protection and deprotection are well known in the art. For example, a number of Protecting Groups and their introduction and removal are described in t.w.greene and p.g.m.wuts, Protecting Groups in Organic Synthesis, Second Edition, Wiley, New York,1991 and references cited therein.
For example, the compounds provided herein can be prepared as follows: the carboxylic acid is reacted with an appropriately substituted amine and the product isolated and purified by standard methods known in the art. Such methods include, but are not limited to, recrystallization, column chromatography, or HPLC. The following synthetic schemes are provided using detailed descriptions for the preparation of representative substituted biaryl amides cited herein. The compounds provided herein can be prepared from starting materials and reagents that are well known or commercially available to those skilled in the art of organic synthesis.
Enantiomerically pure compounds provided herein can be prepared according to any technique known to those skilled in the art. For example, they may be prepared from suitable optically pure precursors by chiral or asymmetric synthesis or obtained by any conventional technique (e.g. by chromatographic resolution using chiral columns, TLC or by preparation of diastereomers, their separation and regeneration of the desired enantiomers). See, for example, "eneromers, racemes and solutions" by j.jacques, a.collets and s.h.wilen (Wiley-Interscience, New York, 1981); s.h.wilen, a.colet and j.jacques, Tetrahedron,2725 (1977); l.l.eliel Stereochemistry of Carbon Compound (McGraw-Hill, NY, 1962); and S.H.Wilen tablets of solving Agents and Optical solutions268(E.L.Eliel ed., Univ.of Notre Dame Press, Notre Dame, IN,1972, Stereochemistry of Organic Compound, Ernest L.Eliel, Samuel H.Wilen and Lewis N.Manda (1994John Wiley & Sons, Inc.) and Stereoselective Synthesis A Practical Approach, Mih ly N gr di (1995VCH Publishers, Inc., NY, NY).
In some embodiments, enantiomerically pure compounds of formula 1 can be obtained by reacting the racemate with a suitable optically active acid or base. Suitable acids or bases include Bighley et al, 1995, Salt Forms of Drugs and Adsorption, in Encyclopedia of Pharmaceutical Technology, vol.13, Swarbrick & Boylan, eds., Marcel Dekker, New York; ten Hoeve & H.Wynberg,1985, Journal of Organic Chemistry50: 4508-4514; dale & Mosher,1973, j.am.chem.soc.95: 512; and CRC Handbook of Optical Resolution via quantitative Salt format, the contents of which are incorporated herein by reference in their entirety.
Enantiomerically pure compounds can also be recovered from crystallized diastereomers or mother liquors, depending on the solubility of the particular acid resolving agent used and the particular acid enantiomer used. The identity and optical purity of the particular compound so recovered can be determined by polarimetry or other analytical methods well known in the art. The diastereomers can then be separated, for example, by chromatography or fractional crystallization and the desired enantiomer regenerated by treatment with a suitable base or acid. The other enantiomers can be obtained in a similar manner from the racemate or from the first separation in the liquid after-treatment.
In some embodiments, enantiomerically pure compounds can be separated from racemic compounds by chiral chromatography. Various chiral columns and eluents for separating enantiomers are available and suitable conditions for separation can be determined empirically by methods well known to those skilled in the art. Typical chiral columns useful for separating enantiomers provided herein include, but are not limited toOB、OB-H、OD、 OD-H、OF、OG、 OJ andOK。
Detailed Description
Various substituted biarylamides can be prepared using the following general methods or synthetic schemes.
General synthetic schemes
In the above synthetic schemes 1 to 11, X and X1Each independently selected from F, Cl, Br, I and OTf; x' is F, Cl, Br, I, Ts or OH; or R is selected from C1-C6Alkyl and benzyl; A. b, W, X', L, R1、R3a、R3b、R4bL and m' are as described herein.
Synthesis of intermediates
Intermediate 1
(S) -2-amino-2- (6- (difluoromethyl) pyridin-3-yl) ethanol
A) 5-bromo-2- (difluoromethyl) pyridine
Treatment of 5-bromo-2-pyridylaldehyde (10g,54 mmol) (org. Lett.2004,6,4905) in anhydrous CH with DAST (9.2g,70mmol) at-78 deg.C2Cl2(100mL) the resulting reaction mixture was warmed to room temperature over a 5h period. After completion of the reaction, the reaction mixture was quenched with ice-cold water and CH2Cl2And (4) extracting. With anhydrous Na2SO4The organic layer was dried and the solvent evaporated in vacuo. By column chromatography (SiO)2,5％Et2O/petroleum ether) to give the title compound (6g, 54% yield). MS 210[ M +1 ]]+；1H-NMR(300MHz,CDCl3):δ8.72(s,1H),7.93(d,1H,J＝8.3Hz),7.54(d,1H,J＝8.3Hz),6.60(t,1H,J＝55.1Hz)。
B)2- (difluoromethyl) -5-vinylpyridines
To potassium vinyltrifluoroborate (3.32g,24.8mmol), PdCl2(0.1g,0.56mmol) and PPh3(0.45g,1.71mmol) in 60mLTHF-H of2Adding Cs to the stirred suspension in O (9:1)2CO3(20.2g,62mmol) and 5-bromo-2- (difluoromethyl) pyridine (4.3g,20.7 mmol). The resulting reaction mixture was heated to 80 ℃ and stirred for 16 h. After completion of the reaction, the reaction mixture was cooled to room temperature, treated with water, and treated with CH2Cl2And (4) extracting. With anhydrous Na2SO4The organic layer was dried and the solvent evaporated in vacuo. By column chromatography (SiO)2,5％Et2O/pentane) to give the title compound (2.46g, 75% yield). MS 156[ M +1 ]]+；1H-NMR(300MHz,CDCl3):δ8.64(s,1H),7.86(d,1H,J＝8.3Hz),7.60(d,1H,J＝8.3Hz),6.74(dd,1H,J＝18,11.2Hz),6.63(t,1H,J＝55.6Hz),5.91(d,1H,J＝18Hz),5.48(d,1H,J＝11.2Hz)。
C) (R) -1- (6- (difluoromethyl) pyridin-3-yl) ethane-1, 2-diol
To an AD-mixture-. beta.at 0 deg.C (21.67g) in 30mL t-BuOH-H2To a stirred suspension in O (1:1) was added 2- (difluoromethyl) -5-vinylpyridine (2.4g,15.5mmol) and the resulting reaction mixture was stirred at 0 ℃ for 5h and then at room temperature. After 16h, use Na2SO3The reaction mixture was worked up in solution and then extracted with EtOAc (3 ×). With anhydrous Na2SO4The combined organic layers were dried and the solvent evaporated in vacuo to give the title compound (2.6g, 87% yield). MS:190[ M +1 ]]+；1H-NMR(300MHz,CDCl3):8.64(s,1H),7.89(d,1H,J＝7.8Hz),7.64(d,1H,J＝7.8Hz),6.64(t,1H,J＝55.4Hz),4.91-4.96(m1H),3.84(dd,1H,J＝11,3.6Hz),3.67(dd,1H,J＝11,7.8Hz)。
D) (R) -2- (tert-butyldimethylsilyloxy) -1- (6- (difluoromethyl) pyridin-3-yl) ethanol
To (R) -1- (6- (difluoromethyl) pyridin-3-yl) ethane-1, 2-diol (2.5g,13.2mmol) in anhydrous CH at room temperature2Cl2To the stirred solution in (1) was added tert-butyldimethylsilyl chloride (2.19g,14.5mmol) and imidazole (0.99g,14.5 mmol). The resulting reaction mixture was stirred for 16 h. After completion of the reaction, the reaction was treated with waterMixture of CH2Cl2And (4) extracting. With anhydrous Na2SO4The organic layer was dried and the solvent evaporated in vacuo. By column chromatography (SiO)220% EtOAc/petroleum ether) to afford the title compound (3g, 75% yield). MS:304[ M +1 ]]+；1H-NMR(300MHz,CDCl3):8.63(s,1H),7.89(d,1H,J＝8.2Hz),7.63(d,1H,J＝8.2Hz),6.64(t,1H,J＝55.4Hz),4.82-4.86(m,1H),3.82(dd,1H,J＝10.1,3.7Hz),3.58(dd,1H,J＝10.1,7.8Hz),0.9(s,9H),0.06(s,6H)。
E) Methanesulfonic acid (R) -2- (tert-butyldimethylsilyloxy) -1- (6- (difluoromethyl) pyridin-3-yl) ethyl ester
To (R) -2- (tert-butyldimethylsilyloxy) -1- (6- (difluoromethyl) pyridin-3-yl) ethanol (3.0g,9.90mmol) in anhydrous CH at 0 deg.C2Cl2Adding MeSO into the stirred solution2Cl (1.2g,10.9mmol) and Et3N (1.3g,12.9 mmol). The resulting reaction mixture was allowed to warm to room temperature over a period of 2 h. After completion of the reaction, the reaction mixture was treated with water and CH2Cl2And (4) extracting. With anhydrous Na2SO4The organic layer was dried and the solvent evaporated in vacuo. By column chromatography (SiO)210% EtOAc/petroleum ether) to afford the title compound (2.26g, 60% yield). MS 381[ M +1 ]]+；1H-NMR(300MHz,CDCl3):8.67(s,1H),7.90(d,1H,J＝8.3Hz),7.68(d,1H,J＝8.3Hz),6.65(t,1H,J＝55.2Hz),5.62-5.66(m,1H),3.98(dd,1H,J＝11.2,6.8Hz),3.87(dd,1H,J＝11.2,4.9Hz),3.028(s,3H),0.87(s,9H),0.04-0.05(m,6H)。
F) (S) -5- (1-azido-2- (tert-butyldimethylsilyloxy) ethyl) -2- (difluoromethyl) pyridine
To a stirred solution of (R) -2- (tert-butyldimethylsilyloxy) -1- (6- (difluoromethyl) pyridin-3-yl) ethyl methanesulfonate (2.24g,5.87mmol) in anhydrous DMF (15mL) was added NaN3(0.45g,7.05mmol) and the resulting reaction mixture was heated at 60 ℃ for 2 h. After completion of the reaction, the reaction mixture was treated with ice-cold water and Et2And (4) extracting. By using anhydrous Na2SO4The organic layer was dried and the solvent evaporated in vacuo to give the title compound (1.63g, 85% yield). MS:328[ M +1 ]]+；1H-NMR(300MHz,CDCl3):8.60(s,1H),7.82(d,1H,J＝8.3Hz),7.64(d,1H,J＝8.3Hz),6.64(t,1H,J＝55.7Hz),4.65-4.70(m1H),3.80-3.86(m,2H),0.89(s,9H),0.04-0.06(m,6H)。
G) (S) -2-azido-2- (6- (difluoromethyl) pyridin-3-yl) ethanol
To a stirred solution of (S) -5- (1-azido-2- (tert-butyldimethylsilyloxy) ethyl) -2- (difluoromethyl) pyridine (1.9g,5.79mmol) in 20mL EtOH at 0 deg.C was added 5mL6N HCl and the resulting reaction mixture was slowly warmed to room temperature over a 2h period. The solvent was evaporated and the residue partitioned between 10% NaHCO3Solution with CH2Cl2In the meantime. With anhydrous Na2SO4The organic layer was dried and the solvent evaporated in vacuo to give the title compound (1.1g, 89% yield). MS:215[ M +1 ]]+；1H-NMR(300MHz,CDCl3):8.63(s,1H),7.86(d,1H,J＝8.3Hz),7.68(d,1H,J＝8.3Hz),6.65(t,1H,J＝55.2Hz),4.75-4.79(m1H),3.70-3.90(m,2H)。
H) (S) -2-amino-2- (6- (difluoromethyl) pyridin-3-yl) ethanol
To a stirred solution of (S) -2-azido-2- (6- (difluoromethyl) pyridin-3-yl) ethanol (1g,4.67mmol) in 15mL THF was added PPh3(2.45g,9.39 mmol). The resulting reaction mixture was stirred for 3h, treated with 0.5mL of water, and stirred for 16 h. After completion of the reaction, the reaction mixture was treated with 2N HCl (15mL) and extracted with EtOAc. By NH3The aqueous layer was treated with water and the volatiles were evaporated to give a residue. By column chromatography (Small neutral Al)2O3Pad, NH3Water/MeOH/CH2Cl21:14:85) to give the title compound (0.74g, 85% yield). MS:189[ M +1 ]]+；.1H-NMR(300MHz,CD3OD):8.72(s,1H),8.09(d,1H,J＝8.3Hz),7.75(d,1H,J＝8.3Hz),6.73(t,1H,J＝55.0Hz),4.39-4.43(m1H),3.90(dd,1H,J＝10.9,4.7Hz),3.80(dd,1H,J＝10.9,6.2Hz)。
Intermediate 2
(S) -2-amino-2- (6- (trifluoromethyl) pyridin-3-yl) ethanol
A)2- (trifluoromethyl) -5-vinylpyridine
2.5M n-butyllithium in hexane (27mL,67.5mmol) was added to methyl triphenyl bromide at-78 deg.C under a nitrogen atmosphere over a 12min period(24.4g,68.4mmol) in THF (150 mL). The reaction was warmed to room temperature to obtain a deep red ylide solution, and a solution of 6- (trifluoromethyl) nicotinaldehyde (10g,57mmol) in THF (50mL) was introduced into the ylide solution with ice cooling. The reaction mixture was warmed to room temperature and stirred for 3 hours. The resulting suspension was heated to 60 ℃ over 30 minutes and then stirred at 60 ℃ for 1 hour. After cooling, the mixture was diluted with water (400mL) and extracted with EtOAc. The organic layer was washed with brine and dried (MgSO)4) And (4) concentrating. The residue was purified by silica gel column (0-10% EtOAc/hexanes) to give the title compound.
B) (R) -1- (6- (trifluoromethyl) pyridin-3-yl) ethane-1, 2-diol
To a 100mL flask were added tert-butanol (49mL), water (49mL) and AD-mixture-beta (13.78 g). Stirring at room temperature yielded two clear phases. The mixture was cooled to 0 ℃ and 2- (trifluoromethyl) -5-vinylpyridine (1.7g,9.8mmol) was added instantaneously. The heterogeneous slurry was stirred vigorously at-20 ℃ overnight. TLC showed the reaction was complete. While stirring the mixture at 0 ℃, solid sodium sulfite (15g,0.12mol) was added and the mixture was warmed to rt and stirred for 1 hour. EtOAc was added to the reaction mixture, and after separation of the layers, the aqueous phase was extracted with EtOAc. With anhydrous MgSO4Drying boxThe combined organic layers were concentrated in vacuo. Passing through a silica gel column (MeOH/CH)2Cl20-10%) to yield the title compound as a white solid.
C) 4-Methylbenzenesulfonic acid (R) -2-hydroxy-2- (6- (trifluoromethyl) pyridin-3-yl) ethyl ester
To (R) -1- (6- (trifluoromethyl) pyridin-3-yl) ethane-1, 2-diol (6.4g,31mmol) and pyridine (20mL) in CH at 0 deg.C2Cl2(200mL) was added p-toluenesulfonyl chloride (7.0g,37mmol) in small portions. The mixture was slowly warmed to rt, stirred for 40 hours, then taken up with CH2Cl2(100mL) dilution. With NaHCO3The organic phase was washed with aqueous solution, brine and dried (Na)2SO4) And concentrated to give the crude title compound.
D) (R) -5- (Oxiran-2-yl) -2- (trifluoromethyl) pyridine
To a stirred solution of 4-methylbenzenesulfonic acid (R) -2-hydroxy-2- (6- (trifluoromethyl) pyridin-3-yl) ethyl ester (1.0g,2.77mmol) in THF (50mL) was added powdered potassium hydroxide (464mg,8.28mmol) dropwise in small portions. The mixture was stirred for 40 minutes and TLC showed the reaction was complete. The mixture was filtered through Celite and the filter cake was washed with acetonitrile (50 mL). The filtrate was carefully concentrated to half volume and the resulting epoxide solution was used directly in the next reaction.
E) (S) -2-azido-2- (6- (trifluoromethyl) pyridin-3-yl) ethanol
To a stirred solution of (R) -5- (oxiran-2-yl) -2- (trifluoromethyl) pyridine (2.72g,14.4mmol) in acetonitrile (200mL) was added lithium perchlorate (20g,0.19mol) and sodium azide (3.7g,57 mmol). The mixture was stirred at 60 ℃ overnight and TLC showed the reaction was complete. After cooling, the mixture was filtered through Celite and the filtrate was concentrated. The residue was treated with water and extracted with EtOAc. The combined organic layers were dried and concentrated. The residue was purified by silica gel column to give the title compound.
F) (S) -2-amino-2- (6- (trifluoromethyl) pyridin-3-yl) ethanol
A mixture of (S) -2-azido-2- (6- (trifluoromethyl) pyridin-3-yl) ethanol (604mg,2.60mmol) in ethyl acetate (28mL) and 10% Pd-C (70mg) in H2Stirring under an atmosphere (1atm) for 1 hr. The catalyst was filtered off and the filtrate was concentrated to give the title compound.1H-NMR(300MHz,CDCl3):8.64(s,1H),7.86(d,1H,J＝8.1Hz),7.60(d,1H,J＝8.1Hz),4.16(m,1H),3.73(dd,1H,J＝10.5,4.2Hz),3.56(dd,1H,J＝10.4,7.2Hz)。
Intermediate 3
C- (1, 1-dioxo-2, 3-dihydro-1H-benzo [ b ] thiophen-6-yl) -methylamine
A) 6-Cyanobenzo [ b ] thiophene-2-carboxylic acid methyl ester
A round-bottomed flask was charged with 3-fluoro-4-formylbenzonitrile (9.00g,60.4mmol), dimethyl sulfoxide (90mL), triphenylphosphine (18.0mL,129mmol), and then methyl 2-mercaptoacetate (5.40mL,60.4 mmol). The reaction mixture was heated at 80 ℃ for 2 hours. After cooling, the reaction mixture was poured into water and the precipitated product was collected by filtration, dried and used in the next step without further purification.
B) 6-cyanobenzo [ b ] thiophene-2-carboxylic acid
A round bottom flask was charged with 6-cyanobenzo [ b ] in water (200mL)]Thiophene-2-carboxylic acid methyl ester (4.00g,17.9mmol), methanol (130mL), and sodium hydroxide (23g,0.58 mol). The reaction was stirred at room temperature for 20 minutes. The reaction was concentrated to half volume and acidified with 6N aqueous HCl. With CHCl3The mixture was extracted with/i-PrOH (90:10) and the organic phase was concentrated under reduced pressure to give the title compound as a brown solid.
C) Benzo [ b ] thiophene-6-carbonitriles
6-cyanobenzo [ b ] thiophene-2-carboxylic acid (1.90g,9.35mmol), 1, 8-diazabicyclo [5.4.0] undec-7-ene (5.6mL,37.4mmol), and N, N-dimethylacetamide (15mL) were added to a microwave vial, and the reaction was subjected to microwave irradiation at 190 ℃ for 1 hour. After cooling, the reaction mixture was poured into 1N aqueous HCl and extracted with ethyl acetate. The organic phase was concentrated under reduced pressure to give the product as a light brown solid.
D)1, 1-dioxo-1H-benzo [ b ] thiophene-6-carbonitrile
Benzo [ b ] thiophene-6-carbonitrile (1.35g,8.48mmol), dichloromethane (270mL) and m-chloroperbenzoic acid (70% purity, 5.85g,23.73mmol) were added in portions to the round-bottom flask over 20 min. The reaction mixture was heated at 45 ℃ overnight. The reaction system produces a 2:1 ratio mixture of sulfoxide and sulfone. The reaction system was diluted with ethyl acetate, quenched with saturated aqueous sodium thiosulfate solution, and stirred for 1 hour. The mixture was then extracted with ethyl acetate and the organic phase was concentrated under reduced pressure. The residue was purified by flash chromatography to give the title compound as a white solid.
E) C- (1, 1-dioxo-2, 3-dihydro-1H-benzo [ b ] thiophen-6-yl) -methylamine
To a hydrogenation par vessel were added 1, 1-dioxo-1H-benzo [ b ] thiophene-6-carbonitrile (0.30g,1.57mmol), ethanol (25mL), and palladium hydroxide (66 mg). The vessel was placed on a shaker for 5h in a 40Psi hydrogen atmosphere. The reaction mixture was filtered through Celite and the filter cake was washed with methanol. The filtrate was concentrated to give the product as a white solid.
Intermediate 4
(S) -2- (tert-butyldimethylsilyloxy) -1- (6-methylpyridin-3-yl) ethylamine
A) (6-methyl-3-pyridyl) methanol
To a solution of methyl 6-methylnicotinate (25g,0.16mol) in diethyl ether (620mL) was added dropwise sodium bis (2-methoxyethoxy) aluminum hydride at room temperature under a nitrogen atmosphere(65 wt.% in toluene, 110mL,0.37 mol). The mixture was then heated to reflux for 1.5 hr. After cooling, the reaction mixture was quenched with water (500mL) at 0 ℃ and the aqueous layer was extracted with EtOAc. With anhydrous MgSO4The combined organic layers were dried, filtered, and concentrated. The residue was purified by silica gel column to give the title compound.
B) 6-methylnicotinaldehydes
Dimethyl sulfoxide (25.3mL,0.357mol) and CH at-78 deg.C under a nitrogen atmosphere2Cl2To a stirred solution (600mL) was added oxalyl chloride (16mL,0.19mol) slowly. After the addition was complete, the mixture was stirred for a further 10 min. To the resulting solution was added dropwise (6-methyl-3-pyridyl) methanol (20g,0.162mol) in CH2Cl2(10mL), and the mixture was stirred at-78 deg.C for 2.5 hr. Triethylamine (110mL,0.82mol) was added slowly at-78 deg.C, and the mixture was allowed to warm slowly to room temperature and stirred for an additional 1 hr. Treating the mixture with water, using CH2Cl2The aqueous phase was extracted (3x500 mL). The combined organic layers were washed with brine, anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by silica gel column to give the title compound.
C) 2-methyl-5-vinylpyridine
To methyl triphenyl bromide at-78 deg.C under nitrogen atmosphere for 1hr period(62.54g,0.175mol) to the suspension in THF (150mL) was added 2.5M n-butyllithium in hexane (69mL,0.17 mol). The mixture was warmed to room temperature to give a dark yellow ylide solution. To the ylide solution was added a solution of 6-methylnicotinaldehyde (18.63g,0.146mol) in THF (50mL) with ice cooling. Will be provided withThe reaction mixture was warmed to room temperature and stirred for 2 hours. The resulting suspension was heated to 60 ℃ over 30 minutes and stirred at 60 ℃ for 1 hour. After cooling, the reaction mixture was diluted with water and extracted with EtOAc. The organic layer was separated, washed with brine and dried (MgSO)4) And (4) concentrating. The residue was purified by silica gel column (0-10% EtOAc/hexanes) to give the title compound.
D) (R) -1- (6-methylpyridin-3-yl) ethane-1, 2-diol
To a 100mL flask was added tert-butanol (480mL), water (480mL) and AD-mixture-beta (138 g). Stirring at room temperature yielded two clear phases, and then the mixture was cooled to 0 ℃. 2-methyl-5-vinylpyridine (11.72g,0.0934mol) was added instantaneously and the heterogeneous slurry was stirred vigorously at-20 ℃ overnight. TLC showed the reaction was complete. While stirring the mixture at 0 ℃, solid sodium sulfite (117.8g,0.934mol) was added and the mixture was warmed to rt and stirred for 1 hour. EtOAc was added and after separation of layers, the aqueous phase was extracted with EtOAc. With anhydrous MgSO4The combined organic layers were dried and concentrated in vacuo to afford the diol as a brown oil.
E) (R) -2- (tert-butyldimethylsilyloxy) -1- (6-methylpyridin-3-yl) ethanol
To (R) -1- (6-methylpyridin-3-yl) ethane-1, 2-diol (13.72g,0.085mol), 1H-imidazole (13.55g,0.197mol) and CH at 0 deg.C2Cl2(180mL) to the stirred mixture was added tert-butyldimethylsilyl chloride (15.31g,0.098 mol). The reaction mixture was stirred at 0 ℃ for 30 minutes, then warmed to room temperature and stirred overnight. The mixture was washed with water (300mL) and CH2Cl2And (4) extracting. The combined organic layers were dried over sodium sulfate and concentrated. The residue was purified by flash chromatography (0-50% EtOAc/hexanes) to give a colorless oil.
F) (S) -5- (1-azido-2- (tert-butyldimethylsilyloxy) ethyl) -2-methylpyridine
To a stirred mixture of (R) -2- (tert-butyldimethylsilyloxy) -1- (6-methylpyridin-3-yl) ethanol (15.6g,0.055mol) and diphenylphosphine azide (62.8mL,0.292mol) in toluene (200mL) at 0 deg.C was added 1, 8-diazabicyclo [5.4.0] undec-7-ene (44.5mL,0.292 mol). The mixture was stirred at 0 ℃ for 30 minutes and then heated at 60 ℃ overnight. After cooling, the mixture was washed with water. The organic layer was dried and concentrated in vacuo. The residue was purified by flash chromatography (0-15% EtOAc/hexanes) to give a colorless oil.
G) (S) -2- (tert-butyldimethylsilyloxy) -1- (6-methylpyridin-3-yl) ethylamine
A mixture of (S) -5- (1-azido-2- (tert-butyldimethylsilyloxy) ethyl) -2-methylpyridine (11.21g,0.036mol), ethyl acetate (400mL) and 10% Pd-C (7g) in H2Stirred under an atmosphere (1atm) for 1 hour. The catalyst was filtered off and the filtrate was concentrated to give the title product as an oil.1H NMR(CD3OD,400MHz):8.39(d,1H,J＝2.0Hz), 7.74(dd,1H,J＝8.0,2.4Hz),7.28(d,J＝8.0Hz,1H),3.98(t,1H,J＝6.0Hz),3.73(d,2H,J＝6.0Hz),2.51(s,3H),0.86(s,9H),-0.02(s,6H)。
Intermediate 5
Imidazo [1,2-a ] pyridin-7-ylmethylamines
A) Imidazo [1,2-a ] pyridin-7-ylmethanols
Lithium tetrahydroaluminate (0.50g,13.2mmol) was slowly added in small portions to imidazo [1,2-a ] ]Pyridine-7-carboxylic acid ethyl ester (2.20g,11.6mmol) in ice-cold THF (150 mL). The mixture was slowly warmed to rt and stirred at rt overnight. The solution was cooled to 0 ℃ and carefully quenched with 1N aqueous HCl (10 mL). Adding solid K2CO3And anhydrous Na2SO4The mixture was stirred at rt for 30 min. The mixture was filtered and the filter cake was washed with THF. The filtrate was concentrated to give the crude product as a solid (1.68g, 80% purity),it was used in the next step without further purification.
B)7- (azidomethyl) imidazo [1,2-a ] pyridines
To imidazo [1,2-a ] at 0 DEG C]Pyridin-7-ylmethanol (80% purity, 1.68g,9.07mmol) in toluene (40mL) and CH2Cl2(40mL) to a stirred solution was added diphenylphosphine azide (3.5mL,16mmol) followed by 1, 8-diazabicyclo [5.4.0 ]]Undec-7-ene (2.5mL,17 mmol). The mixture was stirred at 0 ℃ for 2h, then at rt for 16 h. With water (200mL) and CH2Cl2The reaction mixture was diluted (200 mL). The organic layer was separated, washed with brine and dried (Mg)2SO4) Filtering, and concentrating. The residue was purified by column on silica gel to give the title compound as a colorless oil (1.5g, two-step yield 75%).
C) Imidazo [1,2-a ] pyridin-7-ylmethylamines
Adding 7- (azidomethyl) imidazo [1, 2-a) into a round-bottom flask ]Pyridine (1.5g,8.7mmol), ethyl acetate (200mL) and 10% Pd/C (250 mg). The mixture was stirred in a hydrogen balloon at room temperature for 40 minutes. The mixture was filtered through Celite and the filtrate was concentrated to give the title product (1.2 g).1H NMR(400MHz,DMSO-d6):8.48(d,1H,J＝7.2Hz),7.88(s,1H),7.52(d,1H,J＝1.2Hz),7.51(s,1H),6.89(dd,1H,J＝7.2,1.6Hz),3.83(s,2H)。
Intermediate 6
(4-methyl-3- (methylsulfonyl) phenyl) methylamine
A) Methyl-4-methyl-3- (methylsulfonyl) benzoate
At 0℃Chlorosulfonic acid (170mL,2.57mol) was added to a round bottom flask and 4-methylbenzoic acid (50g,0.37mol) was added in portions. Heating the reaction mixture at 130 deg.CAnd 5 h. The reaction mixture was then poured onto ice and stirred for 10 minutes. The solid formed was filtered, washed with cold water and dried to give 4-methyl-3- (chlorosulfonyl) benzoic acid as a white powder. This compound (80g,0.34mol) was then added portionwise to a round-bottomed flask containing sodium sulfite (200g,2.05mol) and water (80 mL). Aqueous sodium hydroxide (6N) was then added dropwise until the pH of the reaction mixture reached 10 and the reaction was stirred overnight. The reaction was then made acidic (pH2) by the addition of 2N HCl. The solid formed was filtered and dried to give 4-methyl-3-sulfinylbenzoic acid as a white solid. This compound (55g,0.275mol) was added to a round bottom flask containing potassium carbonate (75.8g,0.55mol) and N, N-dimethylformamide (450 mL). Methyl iodide (68.1mL,1.10mol) was added slowly and the reaction was stirred for 4 hours. The reaction mixture was then diluted with water and extracted with EtOAc. The solvent was removed under reduced pressure to give the title compound as a white solid.
B) (4-methyl-3- (methylsulfonyl) phenyl) methanol
To a round-bottom flask was added methyl-4-methyl-3- (methylsulfonyl) benzoate (1.5g,7.0mmol) and methanol (15mL) at 0 ℃. Sodium borohydride (3.7g,100mmol) was added in portions and the reaction was stirred at room temperature for 12 h. The reaction was stopped with ice and extracted with MTBE. The combined organic phases were concentrated under reduced pressure and the residue was purified by flash chromatography to give the title compound.
C)4- (1-azidomethyl) -1-methyl-2- (methylsulfonyl) benzene
To a round-bottom flask were added (4-methyl-3- (methylsulfonyl) phenyl) methanol (6.0g,30.14mmol), toluene (70 mL). Diphenylphosphoryl azide (7.8mL,36.1mmol) and 1, 8-diazabicyclo [5.4.0] undec-7-ene (5.3mL,36.1mmol) were added at 0 deg.C and the reaction mixture was stirred at room temperature overnight. The solvent was removed under reduced pressure, the residue diluted with water and extracted with EtOAc. The combined organic phases were concentrated under reduced pressure and the residue was purified by column chromatography to give the title compound (4g, 62% yield).
D) (4-methyl-3- (methylsulfonyl) phenyl) methylamine
To a round-bottom flask was added 4- (1-azidomethyl) -1-methyl-2- (methylsulfonyl) benzene (5.0g,22.17mmol), THF (40mL), triphenylphosphine (6.4g,24.3mmol) and water (1.6mL,34.2mmol) at 0 deg.C, and the reaction mixture was stirred at room temperature overnight. The solvent was removed, the residue was dissolved in MTBE and 20% HCl di-dropwise added at 0 deg.C The alkane solution, the resulting salt was collected and washed with EtOAc. Then neutralizing the salt with 6N NaOH aqueous solution, and neutralizing with CH2Cl2And (4) extracting. The organic layer was concentrated under reduced pressure to give the title compound as a very thick oil.1H NMR(400MHz,CDCl3):7.98(s,1H),7.50(d,1H,J＝8.0Hz),7.32(d,1H,J＝8.0Hz),3.93(s,2H),3.08(s,3H),2.69(s,3H)。
Intermediate 7
1- (4-chloro-3- (methylsulfonyl) phenyl) ethylamine
A) 4-chloro-3-sulfinylbenzoic acid
At 0℃Chlorosulfonic acid (6.36g,96.0mmol) was added to a round bottom flask and 4-chlorobenzoic acid was added in portions. The reaction mixture was heated at 130 ℃ for 5 h. The reaction mixture was then poured onto ice and stirred for 10 minutes. The solid formed was filtered, washed with cold water and dried to give 4-chloro-3- (chlorosulfonyl) benzoic acid as a white powder. This compound was then added portionwise to a round bottom flask containing sodium sulfite (7.1g,56.4mmol) and water (40 mL). Aqueous sodium hydroxide (6N) was then added dropwise until the pH of the reaction mixture reached 10 and the reaction was stirred overnight. The reaction was then made acidic (pH2) by the addition of 2N HCl. The solid formed was filtered and dried to give the title compound as a white solid.
B) 4-chloro-3- (methylsulfonyl) benzoic acid
To a round bottom flask was added 4-chloro-3-sulfinylbenzoic acid (12g,54.0mmol), potassium carbonate (15.4g,109mmol), and N, N-dimethylformamide (170 mL). Methyl iodide (16.1mL,218mmol) was added slowly and the reaction stirred for 4 hours. The reaction mixture was then diluted with water and extracted with EtOAc. The combined organic layers were concentrated under reduced pressure to give the intermediate methyl-4-chloro-3- (methylsulfonyl) benzoate as an off-white solid. The solid was dissolved in methanol (100mL), 6N sodium hydroxide (50mL) was added, and the reaction was stirred for 3 hours. Methanol was removed under reduced pressure and water was added to the residue. The resulting mixture was then acidified with 2N HCl at 0 ℃, and the solid formed was filtered and dried to give the title compound as a white solid.
C) 4-chloro-N-methoxy-N-methyl-3- (methylsulfonyl) benzamide
To a round bottom flask was added 4-chloro-3- (methylsulfonyl) benzoic acid (7.5g,30.24mmol), dichloromethane (75mL), and N, N-dimethylformamide (0.25 mL). Oxalyl chloride (4.0mL,45.36mmol) was added slowly and the reaction mixture was stirred at room temperature for 1 hour. The solvent was removed to give the intermediate acid chloride, which was dissolved in dichloromethane (50mL), N, O-dimethylhydroxylamine (10.0g,122.9mmol) was added and the reaction mixture was stirred at room temperature overnight. The solvent was removed under reduced pressure and the residue was purified by flash chromatography to give the title compound.
D)1- (4-chloro-3- (methylsulfonyl) phenyl) ethanone
To a round bottom flask was added 4-chloro-N-methoxy-N-methyl-3- (methylsulfonyl) benzamide (18.0g,64.9mmol) and tetrahydrofuran (200mL) at 0 ℃. Methylmagnesium bromide (1M in THF, 78mL,78mmol) was added slowly and the reaction stirred at that temperature for 3 h. The reaction was then quenched with aqueous ammonium chloride solution and extracted with ethyl acetate. The organic layer was concentrated under reduced pressure and the residue was purified by flash chromatography to give the title compound.
E)1- (4-chloro-3- (methylsulfonyl) phenyl) ethanol
To a round bottom flask was added 1- (4-chloro-3- (methylsulfonyl) phenyl) ethanone (12.0g,50.0mmol) and tetrahydrofuran (100mL) at 0 ℃. Sodium borohydride (3.7g,100mmol) was added in portions and the reaction was stirred at room temperature for 12 h. The reaction was stopped with ice and extracted with MTBE. The organic solvent was removed under reduced pressure and the residue was purified by flash chromatography to give the title compound.
F)4- (1-azidoethyl) -1 chloro-2- (methylsulfonyl) benzene
To a round bottom flask was added 1- (4-chloro-3- (methylsulfonyl) phenyl) ethanol (8.5g,32.44mmol) and toluene (100 mL). Diphenylphosphoryl azide (8.4mL,38.93mmol) and 1, 8-diazabicyclo [5.4.0] undec-7-ene (5.8mL,38.93mmol) were added at 0 deg.C and the mixture was stirred at room temperature overnight. The volatile material was removed under reduced pressure, the residue was diluted with water and extracted with EtOAc. The combined organic layers were concentrated under reduced pressure and the residue was purified by column chromatography to give the product (2.4g, 28% yield).
G)1- (4-chloro-3- (methylsulfonyl) phenyl) ethylamine
To a round bottom flask was added 4- (1-azidoethyl) -1 chloro-2- (methylsulfonyl) benzene (2.1g,7.7mmol), THF (20mL), triphenylphosphine (2.4g,8.5mmol) and water (0.6mL,34.2mmol) at 0 deg.C and the reaction was stirred at room temperature overnight. The solvent was removed, the residue was dissolved in MTBE and 20% HCl di-dropwise added at 0 deg.CAnd (3) an alkane solution. The resulting salt was collected. Washed with EtOAc. Then neutralizing the salt with 6N NaOH, and neutralizing with CH2Cl2And (4) extracting. The combined organic layers were concentrated under reduced pressure to give the title compound as a very thick oil.1H NMR(400MHz,CDCl3):8.14(d,1H,J＝2.0Hz),7.63(dd,1H,J＝8.4,2.4Hz),7.52(d,1H,J＝8.0Hz),4.24(q,1H,J＝6.4Hz),3.28(s,3H),1.40(d,3H,J＝6.4Hz)。
Intermediate 8
5- (aminomethyl) -2-chloro-N-cyclopropylbenzenesulfonamide
A) 4-chloro-3- (chlorosulfonyl) benzoic acid methyl ester
To a round bottom flask was slowly added 4-chloro-3- (chlorosulfonyl) benzoic acid (16.0g,63.24mmol) and methanol (100mL) and thionyl chloride (9.3mL,126.5mmol) at 0 ℃. The reaction mixture was then heated to reflux for 1.5 hours. The volatiles were removed under reduced pressure and the residue was diluted with water and extracted with EtOAc. The solvent was removed under reduced pressure to give the title compound as a solid.
B) 4-chloro-3- (N-cyclopropylsulfamoyl) benzoic acid methyl ester
To a round bottom flask was added methyl 4-chloro-3- (chlorosulfonyl) benzoate (12.0g,44.32mmol) and 1, 4-bisAlkane (100 mL). Cyclopropylamine (9.31mL,137.8mmol) was added dropwise and the mixture was stirred at this temperature for 3 h. The solvent was removed and the residue was treated with EtOAc. The organic phase was washed with brine, dried and concentrated to give the title compound (4.5g, 32% yield).
C) 2-chloro-N-cyclopropyl-5- (hydroxymethyl) benzenesulfonamide
Lithium aluminum hydride (2.36g,62.2mmol) was added to a round bottom flask under nitrogen and THF (50mL) was added at 0 ℃. Methyl 4-chloro-3- (N-cyclopropylsulfamoyl) benzoate (4.5g,15.57mmol) in THF (20mL) was added slowly and the mixture stirred at this temperature for 3 h. With 1:1 THF/H2The reaction mixture was quenched with 6N NaOH and filtered through Celite. The filtrate was concentrated under reduced pressure and the residue was purified by flash chromatography to give the title compound (2.5g, 63% yield) as a solid.
D)5- (azidomethyl) -2-chloro-N-cyclopropylbenzenesulfonamide
To a round bottom flask was added 2-chloro-N-cyclopropyl-5- (hydroxymethyl) benzenesulfonamide (2.4g,10mmol) and toluene (30 mL). Diphenylphosphoryl azide (2.19mL,10.1mmol) and 1, 8-diazabicyclo [5.4.0] undec-7-ene (1.31mL,10.1mmol) were added at 0 deg.C and the mixture was stirred at room temperature overnight. The volatiles were removed under reduced pressure and the residue was diluted with water and extracted with EtOAc. The organic solvent was removed under reduced pressure and the residue was purified by column chromatography to give the title compound (1.4g, 53% yield).
D)5- (aminomethyl) -2-chloro-N-cyclopropylbenzenesulfonamide
To a round-bottom flask was added 5- (azidomethyl) -2-chloro-N-cyclopropylbenzenesulfonamide (1.40g,4.87mmol), THF (40mL), triphenylphosphine (1.6g,6.43mmol) and water (0.11mL,6.43mmol) at 0 deg.C and the reaction mixture was allowed to stand at room temperature overnight. The solvent was removed, the residue was dissolved in MTBE and 20% HCl di-dropwise added at 0 deg.CAnd (3) an alkane solution. The resulting salt was collected and washed with EtOAc. Then neutralizing the salt with 6N NaOH, and neutralizing with CH2Cl2And (4) extracting. The combined organic phases were concentrated under reduced pressure to give the title product as a very thick oil.1H NMR(400MHz,DMSO-d6):8.00(s,1H),7.58(s,2H),3.78(s,2H),2.18(m,1H),0.50-0.35(m,4H)。
Synthesis of representative Compounds
Compound 1
(S) -N- (1-hydroxypropan-2-yl) -N-isobutyl-N, 4' -dimethylbiphenyl-3, 5-dicarboxamide
A)4' -Methylbiphenyl-3, 5-dicarboxylic acid dimethyl ester
Under argonTo a mixture of dimethyl 5-bromoisophthalate (2.50g,9.15mmol), p-tolylboronic acid (1.37g,10.1mmol), toluene (50mL), ethanol (10mL), cesium carbonate (3.28g,10.1mmol) and water (5mL) was added tetrakis (triphenylphosphine) -palladium (0) (529mg,0.458mmol) under an atmosphere. The mixture was heated to reflux for 6h, then cooled to room temperature and filtered through Celite. The filtrate was concentrated and the residue was purified by silica gel column (0-50% EtOAc/hexanes) to give a white solid.1H NMR(400MHz,CDCl3):δ8.63(s,1H),8.45(s,2H),7.57(d,2H,J＝8.0Hz),7.29(d,2H,J＝8.0Hz),3.98(s,6H),2.42(s,3H)。
B)5- (methoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid
A mixture of dimethyl 4' -methylbiphenyl-3, 5-dicarboxylate (1.70g,5.98mmol), MeOH (100mL), and 2N aqueous NaOH (8mL) was stirred at 60 deg.C until LC-MS showed almost complete exhaustion of the diester. After cooling, the mixture was concentrated in vacuo and acidified to pH with 1N aqueous HCl<4, extract with EtOAc (100mL x 3). The combined organic layers were washed with brine and dried (Na)2SO4) And concentrated to give a white solid (containing some dicarboxylic acid).
C) (S) -5- (1-hydroxypropan-2-ylcarbamoyl) -4' -methylbiphenyl-3-carboxylic acid methyl ester
To 5- (methoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid (0.60g,2.2mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (470mg,2.4mmol), 1-hydroxybenzotriazole hydrate (340mg,2.2mmol) and CH2Cl2(5mL) to the mixture was added (S) -2-aminopropan-1-ol (180mg,2.4mmol) and N, N-diisopropylethylamine (0.58mL,3.3 mmol). The mixture was stirred at rt overnight, then diluted with EtOAc and with NaHCO3Washed with aqueous solution, brine and dried (Na)2SO4) And (4) concentrating. The residue was purified by silica gel column (0-80% EtOAc/hexanes) to give a white solid. LC-MS:328.2[ M +1 ]]+；1HNMR(400MHz,DMSO-d6):δ8.48(d,1H,J＝8.0Hz),8.40(t,1H,J＝ 1.6Hz),8.37(t,1H,J＝1.6Hz),8.28(t,1H,J＝1.6Hz),7.69(d,2H,J＝8.0Hz),7.34(d,2H,J＝8.0Hz),4.78(t,1H,J＝5.6Hz),4.07(m,1H),3.92(s,3H),3.49(m,1H),3.38(m,1H),2.37(s,3H),1.16(d,3H,J＝6.8Hz)。
D) (S) -5- (1-hydroxypropan-2-ylcarbamoyl) -4' -methylbiphenyl-3-carboxylic acid
A mixture of methyl (S) -5- (1-hydroxypropan-2-ylcarbamoyl) -4' -methylbiphenyl-3-carboxylate (105mg,0.321mmol), lithium hydroxide (9.98mg,0.417mmol), THF (3mL), and water (1mL) was stirred at room temperature for 12 h. LC-MS showed the reaction was complete. The reaction mixture was diluted with water (5mL), acidified to pH 3 with 1N aqueous HCl, and extracted with EtOAc (50mL × 3). The combined organic layers were washed with brine and dried (Na)2SO4) And concentrated to give a white solid. LC-MS 314.4[ M +1 ]]+。
E) (S) -N3- (1-hydroxypropan-2-yl) -N5-isobutyl-N5, 4' -dimethylbiphenyl-3, 5-dicarboxamide
To (S) -5- (1-hydroxypropan-2-ylcarbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (32mg,0.10mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (29mg,0.15mmol), 1-hydroxybenzotriazole hydrate (16mg,0.10mmol), CH2Cl2To a mixture of (3mL) and DMF (0.5mL) were added N-methylisobutylamine (13mg,0.15mmol) and N, N-diisopropylethylamine (0.027mL,0.15 mmol). The mixture was stirred at rt overnight, then diluted with EtOAc and with NaHCO3Washed with aqueous solution, brine and dried (Na)2SO4) And (4) concentrating. By preparative HPLC (100X21.2mm C18 column, 40-80% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:383.3[M+1]+；1H NMR(400MHz,CDCl3) (rotamers) δ 8.00(d,1H, J ═ 5.2Hz),7.66(m,2H),7.50(d,2H, J ═ 8.0Hz),7.27(d,2H, J ═ 8.0Hz),6.52(m,1H),4.30(m,1H),3.81(dd,1H, J ═ 11.2,3.6Hz),3.67(dd,1H, J ═ 11.2,5.6Hz),3.41(d,1H, J ═ 7.2Hz),3.11(d,1H, J ═ 7.6Hz),3.08 and 2.95(s,3H),2.41(s,3H),2.10 and 1.94(m,1H),1.31(d,3H, J ═ 6.01 (s, 6H), 1.77 (d, 8.8, J ═ 8.0Hz), 8.7.6 Hz, 6H, and 7.6 Hz).
Compound 2
(S) -N3- (1-hydroxypropan-2-yl) -N5-isobutyl-4' -methylbiphenyl-3, 5-dicarboxamide
To (S) -5- (1-hydroxypropan-2-ylcarbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (25mg,0.080mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (30mg,0.16mmol), 1-hydroxybenzotriazole hydrate (12mg,0.080mmol), CH2Cl2To a mixture of (3mL) and DMF (0.5mL) was added 2-methyl-1-propylamine (12mg,0.16mmol) and N, N-diisopropylethylamine (0.021mL,0.12 mmol). The mixture was stirred at rt overnight, then diluted with EtOAc and with NaHCO3Washed with aqueous solution, brine and dried (Na)2SO4) And (4) concentrating. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:369.1[M+1]+；1H NMR(400MHz,CDCl3):8.08(m,3H),7.52(m,2H),7.28(m,2H),4.32(m,1H),3.81(dd,1H,J＝11.2,3.6Hz),3.68(dd,1H,J＝11.2,5.6Hz),3.30(d,2H,J＝6.4Hz),2.41(s,3H),1.93(m,1H),1.32(d,3H,J＝6.8Hz),0.99(d,6H,J＝6.4Hz)。
Compound 3
(S) -N- (1-hydroxypropan-2-yl) -4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxamide
A) 3-bromo-5-methylsulfanyl-benzoic acid
A mixture of 3, 5-dibromobenzoic acid (2.5g,8.9mmol), sodium methylsulfide (1.4g,20mmol) and dimethyl sulfoxide (10mL) was sealed in a microwave ovenInside the tube, it was heated in an oil bath at 100 ℃ for 4 hours. TLC showed the reaction was complete. The reaction mixture was poured into water and extracted with EtOAc (3 × 50 mL). The combined organic layers were washed with brine and dried (Na)2SO4) And (4) concentrating. The residue was used in the next step without purification.1H NMR(400MHz,CDCl3):δ7.97(t,1H,J＝1.6Hz),7.87(t,1H,J＝1.6Hz),7.57(t,1H,J＝1.6Hz),2.53(s,3H)。
B) 3-bromo-5- (methylthio) benzoic acid methyl ester
To a stirred mixture of 3-bromo-5-methylsulfanyl-benzoic acid (2.2g,8.9mmol), dichloromethane (100mL) and DMF (5 drops) was added oxalyl chloride (1.0mL,12mmol) at 0 ℃. The mixture was slowly warmed to room temperature and stirred at room temperature overnight. Methanol (5.0mL,120mmol) and N, N-diisopropylethylamine (5.0mL,29mmol) were added and the mixture was stirred at room temperature for 3h, then concentrated. The residue was purified by silica gel column (0-50% EtOAc/hexanes) to give an oil.1H NMR(400MHz,CDCl3):δ7.91(t,1H,J＝1.6Hz),7.81(t,1H,J＝1.6Hz),7.52(t,1H,J＝1.6Hz),3.92(s,3H),2.52(s,3H)。
C) 3-bromo-5- (methylsulfonyl) benzoic acid methyl ester
A mixture of methyl 3-bromo-5- (methylthio) benzoate (1.6g,6.1mmol), m-chloroperbenzoic acid (75% purity, 4.2g,18mmol), and dichloromethane (100mL) was stirred at room temperature overnight. With Na2CO3The reaction mixture was washed with aqueous solution and brine and dried (Na)2SO4) And (4) concentrating. The residue was purified by silica gel column (EtOAc/hexane: 0-70%) to give a white solid.1H NMR(400MHz,CDCl3):δ8.52(s,1H),8.44(s,1H),8.27(s,1H),3.98(s,3H),3.10(s,3H)。
D)4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid methyl ester
To a mixture of methyl 3-bromo-5- (methylsulfonyl) benzoate (0.65g,2.2mmol), p-tolylboronic acid (0.332g,2.44mmol), toluene (10mL), cesium carbonate (0.795g,2.44mmol) and water (1mL) under an argon atmosphere was added tetrakis (triphenylphosphine) -palladium (0) (128mg,0.111 mmol). The mixture was heated under reflux for 15 h. After cooling, the mixture was filtered through Celite and the filter cake was washed with EtOAc. The filtrate was washed with brine and dried (Na)2SO4) And (4) concentrating. The residue was purified by silica gel column (0-100% EtOAc/hexanes) to give a white solid. LC-MS 3.26min,305.3[ M +1 ]]+(weak).
E)4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid
A mixture of methyl 4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxylate (0.55g,1.8mmol), MeOH (50mL), and 2N aqueous NaOH (4mL) was stirred at 50 ℃ for 3 h. After cooling, the mixture was concentrated in vacuo and acidified to pH with 1N aqueous HCl<4, extract with EtOAc (100 mL). The organic layer was washed with brine and dried (Na)2SO4) And concentrated to give a white solid. LC-MS 289.3[ M-1 ]]-；1H NMR(400MHz,CDCl3):δ 8.61(t,1H,J＝1.6Hz),8.59(t,1H,J＝1.6Hz),8.39(t,1H,J＝1.6Hz),7.58(d,2H,J＝8.0Hz),7.33(d,2H,J＝8.0Hz),3.15(s,3H),2.44(s,3H)。
F) (S) -N- (1-hydroxypropan-2-yl) -4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxamide
To 4 '-methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (50mg,0.17mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (36mg,0.19mmol), 1-hydroxybenzotriazole hydrate (26mg,0.17mmol) and CH2Cl2(2mL) to the mixture was added (S) -2-aminopropan-1-ol (19mg,0.26mmol) and N, N-diisopropylethylamine (0.045mL,0.26 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X20.2mm, C18 column; 30-60% MeCN-water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:348.2[M+1]+；1H NMR(400MHz,DMSO-d6):δ8.55(d,1H,J＝8.0Hz),8.42(t,1H,J＝1.6Hz),8.33(t,1H,J＝1.6Hz),8.26(t,1H,J＝1.6Hz),7.75(d,2H,J＝8.0Hz),7.37(d,2H,J＝8.0Hz),4.80(bs,1H),4.08(m,1H),3.49(m,1H),3.39(m,1H),2.50(s,3H),2.38(s,3H),1.17(d,3H,J＝6.8Hz)。
Compound 6
(S) -5- (1-hydroxypropan-2-ylcarbamoyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
A)5- (ethoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid
Dimethyl 4' -methylbiphenyl-3, 5-dicarboxylate (14.3g,50.3mmol) was dissolved in 1, 4-bisAlkane (160mL) and ethanol (200proof, 450 mL). Lithium hydroxide (2.41g,100mmol) was added to the stirred solution and the reaction mixture was stirred at room temperature and monitored by LC-MS. While depleting about 90% of the dimethyl ester to yield predominantly monoethyl ester and some dicarboxylic acid (5-10%), the mixture was cooled, neutralized with 2N aqueous HCl and concentrated in vacuo to about 100 mL. The residue was treated with water (100mL), acidified to pH2-3 with 2N aqueous HCl, and extracted with EtOAc (100mL x 3). The combined organic layers were washed with brine and dried (Na)2SO4) And concentrated to give crude monoethyl ester as a white solid. LC-MS 283.1[ M-1 ]]-。
B) (S) -5- (1-hydroxypropan-2-ylcarbamoyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
To 5- (ethoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid (35mg,0.12mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (47mg,0.25mmol), 1-hydroxybenzotriazole hydrate (19mg,0.12mmol) and CH2Cl2(3mL) to the mixture was added (S) -2-aminopropan-1-ol (10mg,0.14mmol) and N, N-diisopropylethylamine (0.032mL,0.18 mmol). The mixture was stirred at room temperature overnight and then concentrated. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:342.4[M+1]+；1H NMR(400MHz,CDCl3):8.38(t,1H,J＝1.6Hz),8.29(t,1H,J＝1.6Hz),8.24(t,1H,J＝1.6Hz),7.55(d,2H,J＝8.0Hz),7.29(d,2H,J＝8.0Hz),4.44(q,2H,J＝7.2Hz),4.34(m,1H),3.83(dd,1H,J＝11.2,3.6Hz),3.70(dd,1H,J＝11.2,6.0Hz),2.41(s,3H),1.43(t,3H,J＝7.2Hz),1.34(d,3H,J＝6.8Hz)。
Compound 7
(S) -N3- (2-hydroxy-1- (6-methoxypyridin-3-yl) ethyl) -N5-isobutyl-N5, 4' -dimethyl-biphenyl-3, 5-dicarboxamide
A)5- (isobutyl (methyl) carbamoyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
To 5- (ethoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid (260mg,0.91mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (350mg,1.8mmol), 1-hydroxybenzotriazole hydrate (140mg,0.91mmol), CH2Cl2To the mixture (5mL) were added N-methylisobutylamine (120mg,1.4mmol) and N, N-diisopropylethylamine (0.24mL,1.4 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by silica gel column (0-60% EtOAc/hexanes) to give a syrup. LC-MS 354.2[ M +1 ]]+。
B)5- (isobutyl (methyl) carbamoyl) -4' -methylbiphenyl-3-carboxylic acid
A mixture of ethyl 5- (isobutyl (methyl) carbamoyl) -4' -methylbiphenyl-3-carboxylate (165mg, 0.467mmol), lithium hydroxide (55mg,2.3mmol), ethanol (10mL) and water (1mL) was stirred at room temperature for 5 h. LC-MS showed the reaction was complete. The solvent was removed in vacuo and the residue was treated with water, acidified to pH 3 with 1N aqueous HCl and extracted with EtOAc (50mL × 3). The combined organic layers were washed with brine and dried (Na)2SO4) Concentrate to give the product as a white foam. LC-MS 326.0[ M +1 ]]+。
C) (S) -N3- (2-hydroxy-1- (6-methoxypyridin-3-yl) ethyl) -N5-isobutyl-N5, 4' -dimethylbiphenyl-3, 5-dicarboxamide
To a solution of 5- (isobutyl (methyl) carbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (35mg,0.11mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (41mg,0.22mmol), 1-hydroxybenzotriazole hydrate (16mg,0.11mmol), CH2Cl2(3mL) to the mixture was added (S) -2-amino-2- (6-methoxypyridin-3-yl) ethanol (27mg,0.16mmol) (WO2008/130481) and N, N-diisopropylethylamine (28. mu.L, 0.16 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:476.4[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) 8.19(m,2H),7.82-7.70(m,3H),7.59(m,2H),7.30(d,2H, J ═ 8.0Hz),6.80(d,1H, J ═ 8.4Hz),5.19(t,1H, J ═ 6.4Hz),3.94-3.83(m,5H),3.42(d,1H, J ═ 7.6Hz),3.18(d,1H, J ═ 7.6Hz),3.10 and 3.00(s,3H),2.38(s,3H),2.15 and 1.98(m,1H),1.01 and 0.77(d,6H, J ═ 6.8 Hz).
Compound 8
N3-isobutyl-N3, 4' -dimethyl-N5- (1- (pyrazin-2-yl) ethyl) biphenyl-3, 5-dicarboxamide
To 5- (isobutyl (methyl) carbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (21mg,0.064mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (25mg,0.13mmol), 1-hydroxybenzotriazole hydrate (9.9mg,0.064mmol) and CH2Cl2(3mL) to a mixture was added 1- (pyrazin-2-yl) ethylamine (12mg,0.097mmol) and N, N-Diisopropylethylamine (17. mu.L, 0.097 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:431.1[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) δ 8.70(s,1H),8.59(m,1H),8.49(d,1H, J ═ 2.8Hz),8.20(t,1H, J ═ 1.6Hz),7.84-7.75(m,2H),7.62-7.57(m,2H),7.30(d,2H, J ═ 8.0Hz),5.36(q,1H, J ═ 7.2Hz),3.42(d,1H, J ═ 7.6Hz),3.19(d,1H, J ═ 7.6Hz),3.10 and 3.01(s,3H),2.39(s,3H),2.15 and 1.98(m,1H),1.66(d,3H, J ═ 7.2Hz),1.02 (d,6H, J ═ 4H, 6 Hz).
Compound 9
N3- ((6-Chloropyridin-3-yl) methyl) -N5-isobutyl-N5, 4' -dimethylbiphenyl-3, 5-dicarboxamide
To 5- (isobutyl (methyl) carbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (21mg,0.064mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (25mg,0.13mmol), 1-hydroxybenzotriazole hydrate (9.9mg,0.064mmol) and CH2Cl2(3mL) to a mixture was added 2-chloro-5-aminomethylpyridine (14mg,0.097mmol) and N, N-diisopropylethylamine (17. mu.L, 0.097 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 40-80% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:450.4[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) δ 8.39(d,1H, J ═ 2.0Hz),8.18(m,1H),7.85(dd,1H, J ═ 8.0,2.4Hz),7.82-7.75(m,2H),7.58(m,2H),7.44(d,1H, J ═ 8.0Hz),7.30(d,2H, J ═ 7.6Hz),4.61(s,2H),3.42(d,1H, J ═ 7.6Hz),3.19(d,1H, J ═ 7.2Hz),3.10 and 3.01(s,3H),2.38(s,3H),2.15 and 1.98(m,1H),1.01 and 0.77(d,6H,J＝6.8Hz)。
Compound 10
(R) -N3-isobutyl-N3, 4' -dimethyl-N5- (1- (6- (trifluoromethyl) pyridin-3-yl) ethyl) biphenyl-3, 5-dicarboxamide
To 5- (isobutyl (methyl) carbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (21mg,0.064mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (25mg,0.13mmol), 1-hydroxybenzotriazole hydrate (9.9mg,0.064mmol) and CH2Cl2(3mL) to the mixture was added (R) -1- (6- (trifluoromethyl) pyridin-3-yl) ethylamine (18mg,0.097mmol) (WO2008/130481) and N, N-diisopropylethylamine (17. mu.L, 0.097 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 40-80% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:498.6[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) δ 8.77(d,1H, J ═ 1.6Hz),8.19(t,1H, J ═ 1.6Hz),8.07(dd,1H, J ═ 8.0,2.0Hz),7.83-7.75(m,3H),7.59(m,2H),7.30(d,2H, J ═ 8.4Hz),5.36(q,1H, J ═ 6.4Hz),3.42(d,1H, J ═ 7.6Hz),3.18(d,1H, J ═ 7.6Hz),3.10 and 3.00(s,3H),2.39(s,3H),2.15 and 1.98(m,1H),1.66(d,3H, J ═ 7.2), 1.01 and 0.77(d,6H, 6 Hz).
Compound 11
N3- (imidazo [1,2-a ] pyridin-7-ylmethyl) -N5-isobutyl-N5, 4' -dimethylbiphenyl-3, 5-dicarboxamide
To 5- (isobutyl (methyl) carbamoylYl) -4 '-methylbiphenyl-3-carboxylic acid (15mg,0.046mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (18mg,0.092mmol), 1-hydroxybenzotriazole hydrate (7.0mg,0.046mmol) and CH2Cl2(2mL) to the mixture was added imidazo [1,2-a ]]Pyridin-7-ylmethylamine (10mg,0.069mmol) and N, N-diisopropylethylamine (12. mu.L, 0.069 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 40-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:455.4[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) δ 8.40(d,1H, J ═ 7.2Hz),8.22(s,1H),7.87-7,76(m,3H),7.62-7.57(m,2H),7.52(d,1H, J ═ 1.6Hz),7.49(s,1H),7.30(d,2H, J ═ 8.0Hz),6.96(dd,1H, J ═ 7.2,1.6Hz),4.66(s,2H),3.43(d,1H, J ═ 8.0Hz),3.20(d,1H, J ═ 7.6Hz),3.10 and 3.02(s,3H),2.39(s,3H),2.15 and 1.98(m,1H),1.01 and 0.78(d,6H, 4.78 Hz).
Compound 12
N3-isobutyl-N3, 4' -dimethyl-N5- ((2-methylpyrimidin-5-yl) methyl) biphenyl-3, 5-dicarboxamide
To 5- (isobutyl (methyl) carbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (15mg,0.046mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (18mg,0.092mmol), 1-hydroxybenzotriazole hydrate (7.0mg,0.046mmol) and CH2Cl2(2mL) to the mixture was added (2-methylpyrimidin-5-yl) methylamine (8.5mg,0.069mmol) (WO2008/130481) and N, N-diisopropylethylamine (12. mu.L, 0.069 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mmC18 column, 40-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:431.3[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) δ 8.72(s,2H),8.17(s,1H),7.82-7.75(m,2H),7.60-7.55(m,2H),7.30(d,2H, J ═ 8.4Hz),4.59(s,2H),3.42(d,1H, J ═ 7.6Hz),3.18(d,1H, J ═ 7.6Hz),3.10 and 3.00(s,3H),2.67(s,3H),2.38(s,3H),2.15 and 1.98(m,1H),1.01 and 0.77(d,6H, J ═ 6.4 Hz).
Compound 13
(R) -N3- (3-hydroxy-1- (6-methylpyridin-3-yl) propyl) -N5-isobutyl-N5, 4' -dimethylbiphenyl-3, 5-dicarboxamide
To 5- (isobutyl (methyl) carbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (15mg,0.046mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (18mg,0.092mmol), 1-hydroxybenzotriazole hydrate (7.0mg,0.046mmol) and CH2Cl2(2mL) to the mixture was added (R) -3-amino-3- (6-methylpyridin-3-yl) propan-1-ol (11mg,0.069mmol) (WO2008/130481) and N, N-diisopropylethylamine (12. mu.L, 0.069 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by preparative HPLC (100X21.2mm C18 column, 40-70% MeCN/water [10mM Et2NH]) A white foam was obtained.
LC-MS:474.6[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) δ 8.47(d,1H, J ═ 1.6Hz),8.15(s,1H),7.80-7.73(m,3H),7.60-7.55(m,2H),7.30(d,3H, J ═ 8.0Hz),5.32(m,1H),3.72-3.57(m,2H),3.42(d,1H, J ═ 7.6Hz),3.17(d,1H, J ═ 7.6Hz),3.09 and 3.00(s,3H),2.51(s,3H),2.38(s,3H),2.24-1.94(m,3H),1.01 and 0.77(d,6H, J ═ 6.4 Hz).
Compound 14
(R) -N3-isobutyl-N3, 4' -dimethyl-N5- (1- (2-methylpyrimidin-5-yl) ethyl) biphenyl-3, 5-dicarboxamide
To 5- (isobutyl (methyl) carbamoyl) -4 '-methylbiphenyl-3-carboxylic acid (15mg,0.046mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (18mg,0.092mmol), 1-hydroxybenzotriazole hydrate (7.0mg,0.046mmol) and CH2Cl2(2mL) to the mixture was added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (9.5mg,0.069 mmol) (WO2008/130481) and N, N-diisopropylethylamine (12. mu.L, 0.069 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 40-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:445.6[M+1]+；1H NMR(400MHz,CD3OD) (rotamers) δ 8.77(s,2H),8.17(t,1H, J ═ 1.6Hz),7.81-7.74(m,2H),7.61-7.55(m,2H),7.30(d,2H, J ═ 8.4Hz),5.27(q,1H, J ═ 7.2Hz),3.42(d,1H, J ═ 7.6Hz),3.18(d,1H, J ═ 7.6Hz),3.09 and 3.00(s,3H),2.67(s,3H),2.39(s,3H),2.15 and 1.98(m,1H),1.65(d,3H, J ═ 6.8Hz),1.01 and 0.77(d,6H, J ═ 6.8 Hz).
Compound 15
4' -methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) biphenyl-3-carboxylic acid ethyl ester
To crude 5- (ethoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid (450mg,1.6mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (610mg,3.2mmol), 1-hydroxybenzotriazole hydrate (240mg,1.6mmol), CH2Cl2(5mL) to a mixture was added (2-methylpyrimidin-5-yl) methylamine (290mg,2.4mmol) and N, N-diisopropylethylamine (0.41mL,2.4 mmol). The reaction mixture was stirred at room temperature overnight and then with NaHCO3Aqueous and EtOAc (100 mL). The organic layer was separated, washed with brine, dried over anhydrous sodium sulfate and concentrated in vacuo. Passing through a silica gel column (0-20% MeOH/CH)2Cl2) The residue was purified to give the title product as a white foam.
LC-MS:390.4[M+1]+；1H NMR(400MHz,CDCl3):8.73(s,2H),8.40(t,1H,J＝1.6Hz),8.29(t,1H,J＝1.6Hz),8.26(t,1H,J＝1.6Hz),7.54(d,2H,J＝8.0Hz),7.28(d,2H,J＝8.0Hz),6.72(bs,1H),4.67(d,2H,J＝6.0Hz),4.43(q,2H,J＝7.2Hz),2.76(s,3H),2.41(s,3H),1.42(t,3H,J＝7.2Hz)。
Compound 16
4' -methyl-N3, N5-bis ((2-methylpyrimidin-5-yl) methyl) biphenyl-3, 5-dicarboxamide
The title compound was isolated from the preparation of compound 15 as a white foam because the crude 5- (ethoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid contained some 4' -methylbiphenyl-3, 5-dicarboxylic acid.
LC-MS:467.4[M+1]+；1H NMR(400MHz,CDCl3):8.66(s,4H),8.09(s,2H),8.06(s,1H),7.46(d,2H,J＝8.4Hz),7.25(d,2H,J＝8.8Hz),6.95(t,2H,J＝5.6Hz),4.61(d,4H,J＝6.0Hz),2.71(s,6H),2.39(s,3H)。
Compound 17
5- (3, 3-difluoroazetidine-1-carbonyl) -4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) biphenyl-3-carboxamide
To 4' -methyl-5- ((2-methylpyrimidin-5-yl) methylamineCarbamoyl) biphenyl-3-carboxylic acid (11mg,0.030mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (12mg,0.061mmol), 1-hydroxybenzotriazole hydrate (4.7mg,0.030mmol), CH2Cl2To the mixture (2mL) was added 3, 3-difluoroazetidine (4.2mg,0.046mmol) and N, N-diisopropylethylamine (11. mu.L, 0.061 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:437.3[M+1]+；1H NMR(400MHz,CD3OD):δ8.73(s,2H),8.26(t,1H,J＝1.6Hz),8.06(m,2H),7.60(d,2H,J＝8.4Hz),7.31(d,2H,J＝8.0Hz),4.77(bs,2H),4.59(s,2H),4.56(bs,2H),2.67(s,3H),2.38(s,3H)。
Compound 18
4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) -5- (piperidine-1-carbonyl) biphenyl-3-carboxamide
To 4 '-methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (11mg,0.030mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (12mg,0.061mmol), 1-hydroxybenzotriazole hydrate (4.7mg,0.030mmol) and CH2Cl2(2mL) was added piperidine (3.9mg,0.046mmol) and N, N-diisopropylethylamine (11. mu.L, 0.061 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:429.3[M+1]+；1H NMR(400MHz,CD3OD):δ8.72(s,2H),8.17(t,1H,J＝1.6Hz),7.80(t,1H,J＝1.6Hz),7.78(t,1H,J＝1.6Hz),7.58(d,2H,J＝8.0Hz),7.30(d,2H,J＝8.0Hz),4.58(s,2H),3.74(bs,2H),3.41(bs,2H),2.67(s,3H),2.38(s,3H),1.71(bs,4H),1.56(bs,2H)。
Compound 19
5- (azepane-1-carbonyl) -4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) biphenyl-3-carboxamide
To 4 '-methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (11mg,0.030mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (12mg,0.061mmol), 1-hydroxybenzotriazole hydrate (4.7mg,0.030mmol) and CH2Cl2To the mixture (2mL) was added hexahydro-1H-azepine (4.5mg,0.046mmol) and N, N-diisopropylethylamine (11. mu.L, 0.061 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:443.5[M+1]+；1H NMR(400MHz,CD3OD):δ8.72(s, 2H),8.17(t,1H,J＝1.6Hz),7.80(t,1H,J＝1.6Hz),7.77(t,1H,J＝1.6Hz),7.58(d,2H,J＝8.4Hz),7.30(d,2H,J＝8.0Hz),4.58(s,2H),3.70(t,2H,J＝6.0Hz),3.45(t,2H,J＝5.6Hz),2.67(s,3H),2.38(s,3H),1.90-1.80(m,2H),1.73-1.58(m,6H)。
Compound 20
4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To 4' -methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (11mg,0.030mmol), N- (3-dimethyl) Aminopropyl) -N' -ethylcarbodiimide hydrochloride (12mg,0.061mmol), 1-hydroxybenzotriazole hydrate (4.7mg,0.030mmol) and CH2Cl2To the mixture (2mL) was added pyrrolidine (3.2mg,0.046mmol) and N, N-diisopropylethylamine (11. mu.L, 0.061 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:415.2[M+1]+；1H NMR(400MHz,DMSO-d6):9.27(t,1H,J＝5.6Hz),8.68(s,2H),8.20(t,1H,J＝1.6Hz),7.93(t,1H,J＝1.4Hz),7.90(t,1H,J＝1.5Hz),7.67(d,2H,J＝8.1Hz),7.31(d,2H,J＝8.0Hz),4.49(d,2H,J＝5.6Hz),3.50(t,2H,J＝6.7Hz),3.42(t,2H,J＝6.4Hz),2.60(s,3H),2.36(s,3H),2.00-1.78(m,4H)。
Compound 21
4' -methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) biphenyl-3-carboxylic acid
A mixture of 4' -methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid ethyl ester (275mg,0.706mmol), lithium hydroxide (85mg,3.5mmol), THF (10mL), and water (1mL) was stirred at room temperature for 5 h. LC-MS showed the reaction was complete. The solvent was removed in vacuo and the residue was treated with water and acidified to pH 4-5 with 1N aqueous HCl. The precipitate was collected by filtration and dried to give the title compound as a white solid. The filtrate was extracted with EtOAc (100 mL). The organic layer was washed with water and concentrated to give some more product.
LC-MS:362.4[M+1]+；1H NMR(400MHz,DMSO-d6):δ13.33(s,1H),9.36(t,1H,J＝5.6Hz),8.69(s,2H),8.41(t,1H,J＝1.6Hz),8.36(t,1H,J＝1.6Hz),8.29(t,1H,J＝1.6Hz),7.67(d,2H,J＝8.0Hz),7.33(d,2H,J＝8.0Hz),4.50(d,2H,J＝5.6Hz),2.60(s,3H),2.37(s,3H)。
Compound 22
4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) -5- (methylsulfonyl) biphenyl-3-carboxamide
To 4 '-methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (45mg,0.15mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (59mg,0.31mmol), 1-hydroxybenzotriazole hydrate (24mg,0.15mmol) and CH2Cl2(3mL) to a mixture was added (2-methylpyrimidin-5-yl) methylamine (29mg,0.23mmol) and N, N-diisopropylethylamine (54. mu.L, 0.31 mmol). The mixture was stirred at room temperature overnight and then concentrated. By preparative HPLC (100X20.2mm, C18 column; 30-60% MeCN-water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:396.3[M+1]+；1H NMR(400MHz,DMSO-d6):δ9.45(t,1H,J＝5.6Hz),8.70(s,2H),8.45(t,1H,J＝1.6Hz),8.33(t,1H,J＝1.6Hz),8.29(t,1H,J＝1.6Hz),7.75(d,2H,J＝8.4Hz),7.36(d,2H,J＝8.0Hz),4.52(d,2H,J＝5.6Hz),2.60(s,3H),2.50(s,3H),2.38(s,3H)。
Compound 28
3- (5-methylpyridin-2-yl) -N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidine-1-carbonyl) benzamide
A) 3-bromo-5- (methoxycarbonyl) benzoic acid
To a round bottom flask was added dimethyl 5-bromoisophthalate (0.50g,1.8mmol), barium hydroxide octahydrate (0.43g,1.4mmol) and methanol (10 mL). The mixture was stirred at room temperatureOvernight. HCl (2N in ether, 10mL) was added and the volatiles were removed under reduced pressure. The residue was purified by flash chromatography to afford the desired product as a white solid.1H NMR(400MHz,DMSO-d6):13.70(Br、1H),8.42(t,J＝1.5Hz,1H),8.31-8.24(m,2H),3.90(s,3H)。
B) 3-bromo-5- (pyrrolidine-1-carbonyl) benzoic acid methyl ester
To a mixture of 3-bromo-5- (methoxycarbonyl) benzoic acid (1.8g,5.6mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (2.1g,11mmol), 1-hydroxybenzotriazole hydrate (1.7g,11mmol), dichloromethane (20mL) was added pyrrolidine (0.58g,8.2mmol) and N, N-diisopropylethylamine (1.9mL,11 mmol). The mixture was stirred at room temperature overnight and then diluted with dichloromethane (200 mL). By BaHCO3Aqueous solution and K2HPO4The organic phase was washed with aqueous solution, brine and Na2SO4Drying and concentrating. The residue was purified by flash chromatography (silica gel column, 0-100 EtOAc/hexanes) to give the desired product as a clear oil.1H NMR(400MHz,DMSO-d6):8.12(t,J＝1.8Hz,1H),7.80-7.98(m,2H),3.88(s,3H),3.47(t,J＝6.7Hz,2H),3.36(t,J＝6.5Hz,2H),1.89-1.80(m,4H)。
C) 3-bromo-5- (pyrrolidine-1-carbonyl) benzoic acid
To a round bottom flask was added methyl 3-bromo-5- (pyrrolidine-1-carbonyl) benzoate (0.86g,2.5mmol), lithium hydroxide (0.071g,3.0mmol), methanol (5mL) and water (5 mL). The mixture was stirred at room temperature for 2 h. Methanol was removed under reduced pressure. The aqueous layer was acidified to pH 1 with 1N aqueous HCl and then with CH2Cl2And (4) extracting. With Na2SO4The separated organic phase was dried, filtered and concentrated to dryness to give the desired product as a white solid.1H NMR(400MHz,DMSO-d6):13.54(Br、1H),8.09(t,J＝1.6Hz,1H),7.98(t,J＝1.5Hz,1H),7.94(t,J＝1.8Hz,1H),3.47(t,J＝6.7Hz,2H),3.37(t,J＝6.5Hz,2H),1.90-1.80(m,4H)。
D) 3-bromo-N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidine-1-carbonyl) benzamide
To a round bottom flask was added 3-bromo-5- (pyrrolidine-1-carbonyl) benzoic acid (340mg,1.0mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (0.39g,2.0mmol), 1-hydroxybenzotriazole (0.30g,2.2mmol), N, N-diisopropylethylamine (0.2g,1.5mmol), dichloromethane (10mL), 4-dimethylaminopyridine (5mg,0.04mmol) and (2-methylpyrimidin-5-yl) methylamine (240mg,1.5 mmol). The mixture was stirred at room temperature overnight and then with CH2Cl2And (6) diluting. With Na2HPO4The separated organic phase was washed with aqueous solution, brine and Na2SO4Drying and concentrating. The residue was purified by flash chromatography to give the desired product as an oil. LC-MS 404.8[ M +1 ]]+；1H NMR(400MHz,CDCl3):8.68(s,2H),7.95(t,J＝1.6Hz,1H),7.76(t,J＝1.3Hz,1H),7.74-7.71(m,1H),7.61(t,J＝1.5Hz,1H),4.57(d,J＝5.8Hz,2H),3.56(t,J＝6.8Hz,2H),3.37(t,J＝6.4Hz,2H),2.73(s,3H),1.96-1.89(m,4H)。
E)3- (5-methylpyridin-2-yl) -N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidine-1-carbonyl) benzamide
A mixture of 3-bromo-N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidine-1-carbonyl) benzamide (25mg,0.059mmol), 5-methyl-2- (tributylstannyl) pyridine (40mg,0.10mmol), tetrakis (triphenylphosphine) palladium (0) (5mg,0.0043mmol) and toluene (1mL) was subjected to microwave irradiation at 120 ℃ for 1 hour under a nitrogen atmosphere. After cooling, the mixture was diluted with water and extracted with EtOAc. With Na2SO4The combined organic layers were dried and concentrated. The residue was purified by flash chromatography to give the title compound as a white solid.
LC-MS:416.3[M+1]+；1H NMR(400MHz,DMSO-d6):9.29(t,J＝5.4Hz,1H),8.67(s,2H),8.62(t,J＝1.7Hz,1H),8.54(d,J＝2.2Hz,1H),8.32(t,J＝1.6Hz,1H),8.01-7.99(m,2H),7.75(dd,J＝1.7,8.0Hz,1H),4.49(d,J＝5.6Hz,2H),3.50(t,J＝6.7Hz,2H),3.41(t,J＝6.3Hz,2H),2.60(s,3H),2.36(s,3H),1.91-1.81(m,4H)。
Compound 29
5- (3-hydroxyazetidine-1-carbonyl) -4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) biphenyl-3-carboxamide
A)5- (3-Hydroxyazetidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
To 5- (ethoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid (2.0g,7.0mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (2.7g,14mmol), 1-hydroxybenzotriazole hydrate (1.1g,7.0mmol) and CH2Cl2(50mL) was added 3-hydroxyazetidine hydrochloride (1.2g,10mmol) and N, N-diisopropylethylamine (3.7mL,21 mmol). The mixture was stirred at room temperature for 5h, then brine and Na2CO3Washed with an aqueous solution and dried (Na)2SO4) And (4) concentrating in vacuum. The residue was purified by silica gel column (0-10% MeOH/CH)2Cl2) To obtain syrup. LC-MS 340.4[ M +1 ]]+；1H NMR(400MHz,CDCl3):8.34(t,1H,J＝1.6Hz),8.19(t,1H,J＝1.6Hz),8.04(t,1H,J＝1.6Hz),7.52(d,2H,J＝8.4Hz),7.28(d,2H,J＝8.4Hz),4.75(bs,1H),4.50(m,2H),4.41(q,2H,J＝7.2Hz),4.25(bs,1H),4.10(bs,1H),2.82(bs,1H),2.41(s,3H),1.42(t,3H,J＝7.2Hz)。
B)5- (3-hydroxyazetidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylic acid
A mixture of ethyl 5- (3-hydroxyazetidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylate (2.0g,5.9mmol), lithium hydroxide (0.56g,24mmol), methanol (100mL) and water (10mL) was stirred at rt for 4 h. LC-MS showed the reaction was complete. The solvent was removed in vacuo and the residue was treated with water, acidified to pH2-3 with 1N aqueous HCl and extracted with EtOAc (50mL x 3). The combined organic layers were washed with brine and dried (Na)2SO4) And concentrated to give a white solid. LC-MS 312.4[ M +1 ]]+；1H NMR(400MHz,DMSO-d6):13.30(bs,1H),8.26(s,1H),8.09(s,1H),8.03(s,1H),7.64(d,2H,J＝8.0Hz),7.32(d,2H,J＝8.0Hz),5.79(bs,1H),4.52(s,2H),4.30(bs,1H),4.10(bs,1H),3.84(d,1H,J＝9.6Hz),2.37(s,3H)。
C)5- (3-hydroxyazetidine-1-carbonyl) -4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) biphenyl-3-carboxamide
To 5- (3-hydroxyazetidine-1-carbonyl) -4 '-methylbiphenyl-3-carboxylic acid (350mg,1.1mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (430mg,2.2mmol), 1-hydroxybenzotriazole hydrate (170mg,1.1mmol), CH2Cl2(50mL) to a mixture was added (2-methylpyrimidin-5-yl) methylamine (210mg,1.7mmol) and N, N-diisopropylethylamine (0.39mL,2.2 mmol). The mixture was stirred at room temperature for 4h, then water and Na were used2CO3Washed with an aqueous solution and dried (Na)2SO4) And (4) concentrating in vacuum. By preparative HPLC (100X21.2mm C18 column, 20-60% MeCN/water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:417.5[M+1]+；1H NMR(400MHz,CD3OD):8.73(s,2H),8.22(t,1H,J＝1.6Hz),8.03(t,1H,J＝1.6Hz),8.01(t,1H,J＝1.6Hz),7.59(d,2H,J＝8.0Hz),7.30(d,2H,J＝8.0Hz),4.66-4.55(m,4H),4.42(m,1H),4.18(m,1H),3.97(m,1H),2.67(s,3H),2.39(s,3H)。
Compound 31
4' -methyl-5- (2-methylaziridine-1-carbonyl) -N- ((2-methylpyrimidin-5-yl) methyl) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (11mg,0.030mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (12mg,0.061mmol), 1-hydroxybenzotriazole hydrate (4.7mg,0.030mmol),CH2Cl2(2mL) to the mixture was added 2-methyl-aziridine (3.5mg,0.061mmol) and N, N-diisopropylethylamine (11. mu.L, 0.061 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) A white foam was obtained.
LC-MS:401.3[M+1]+；1H NMR(400MHz,CD3OD) 8.74(s,2H),8.28-8.13(m,3H),7.62(m,2H),7.31(d,2H, J-8.4 Hz),5.02(m,1H),4.71(m,1H),4.60(s,2H),3.84(m,1H),2.67(s,3H),2.40(s,3H),1.50 and 1.45(d,3H, J-6.8 Hz).
Compound 34
5- (3, 3-difluoropyrrolidine-1-carbonyl) -4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) biphenyl-3-carboxamide
To 4 '-methyl-5- ((2-methylpyrimidin-5-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (11mg,0.030mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (12mg,0.061mmol), 1-hydroxybenzotriazole hydrate (5mg,0.030mmol), CH2Cl2(2mL) to the mixture was added 3, 3-difluoropyrrolidine hydrochloride (9mg,0.061mmol) and N, N-diisopropylethylamine (21. mu.L, 0.12 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-70% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:451.2[M+1]+；1H NMR(400MHz,CD3OD):8.73(s,2H),8.22(s,1H),7.94(s,2H),7.60(d,2H,J＝8.0Hz),7.30(d,2H,J＝8.0Hz),4.59(s,2H),4.03-3.75(m,4H),2.67(s,3H),2.45(m,2H),2.38(s,3H)。
Compound 37
4' -methyl-5- (methylsulfonyl) -N- ((6- (trifluoromethyl) pyridin-3-yl) methyl) biphenyl-3-carboxamide
To 4 '-methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (40mg,0.14mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (53mg,0.28mmol), 1-hydroxybenzotriazole hydrate (21mg,0.14mmol) and CH2Cl2(3mL) to a mixture was added C- (6-trifluoromethyl-pyridin-3-yl) -methylamine (36mg,0.21mmol) and N, N-diisopropylethylamine (48. mu.L, 0.28 mmol). The mixture was stirred at room temperature overnight and then concentrated. By preparative HPLC (100X20.2mm, C18 column; 40-80% MeCN-water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:449.4[M+1]+；1H NMR(400MHz,CDCl3):8.77(s,1H),8.35(t,1H,J＝1.6Hz),8.28(t,1H,J＝1.6Hz),8.21(t,1H,J＝1.6Hz),7.95(d,1H,J＝8.0Hz),7.70(d,1H,J＝8.0Hz),7.55(d,2H,J＝8.0Hz),7.31(d,2H,J＝8.0Hz),6.89(t,1H,J＝6.0Hz),4.78(d,2H,J＝6.0Hz),3.12(s,3H),2.42(s,3H)。
Compound 39
2 '-cyano-4' -methyl-N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To a Parr pressure reactor was added 3-bromo-N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidine-1-carbonyl) benzamide (240mg,0.48mmol), 5-methyl-2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzonitrile (160mg,0.58mmol) (WO2008/130481), toluene (5mL), ethanol (1mL), cesium carbonate (170mg,0.52mmol), and water (0.5 mL). The mixture was degassed, purged several times with nitrogen, then tetrakis (triphenylphosphine) palladium (0) (28mg,0.024mmol) was added. The tube was sealed and the mixture was heated at 90 ℃ overnight. After cooling, the mixture was filtered through Celite and the filtrate was concentrated. The residue was purified by flash chromatography, then by preparative HPLC to give the desired product as a white solid.
LC-MS:440.3[M+1]+；1H NMR(400MHz,DMSO-d6):9.26(t,J＝5.9Hz,1H),8.67(s,2H),8.10-8.07(m,2H),7.83(dd,J＝1.6,9.7Hz,2H),7.62(t,J＝8.0Hz,2H),4.48(d,J＝5.7Hz,2H),3.51-3.43(m,4H),2.59(s,3H),2.41(s,3H),1.90-1.82(m,4H)。
Compound 45
N- (1- (4-chloro-3- (methylsulfonyl) phenyl) ethyl) -4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
A)5- (Chlorocarbonyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
To 5- (ethoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid (5.0g,18mmol), DMF (0.05mL) and CH at 0 deg.C2Cl2To a stirred solution (150mL) was added oxalyl chloride (2.23mL,26.4 mmol). The mixture was stirred at rt for 6h, then concentrated in vacuo to afford the crude acid chloride for the next reaction.
B)4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid ethyl ester
To a stirred solution of ethyl 5- (chlorocarbonyl) -4' -methylbiphenyl-3-carboxylate (5.5g,18mmol) in dichloromethane (100mL) was slowly added pyrrolidine (2.6g,36mmol) and triethylamine (7.6mL,54mmol) at 0 ℃. The mixture was stirred at room temperature for 2h, then brine, Na2CO3Aqueous solution and water wash. Drying the separated organic phase (Na)2SO4) And (4) concentrating. The residue was purified by silica gel column (30-100% EtOAc/hexanes),the desired compound is obtained.
C)4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid
A mixture of ethyl 4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylate (3.94g,11.7mmol), lithium hydroxide (0.65g,27.1mmol), MeOH (250mL), and water (40mL) was stirred at rt for 20 hr. LC-MS showed the reaction was complete. The solvent was removed in vacuo and the residue was treated with water and acidified to pH 2-3 with 1N aqueous HCl. The aqueous phase was extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with water, brine, dried and concentrated to give the product as a white solid. LC-MS 310.4[ M +1 ]]+；1H NMR(400MHz,DMSO-d6):13.29(s,1H),8.21(t,1H,J＝1.6Hz),7.99(t,1H,J＝1.6Hz),7.98(t,1H,J＝1.5Hz),7.64(d,2H,J＝8.2Hz),7.31(d,2H,J＝7.9Hz),3.50(t,2H,J＝6.7Hz),3.43(t,2H,J＝6.5Hz),2.36(s,3H),1.98-1.78(m,4H)。
D) N- (1- (4-chloro-3- (methylsulfonyl) phenyl) ethyl) -4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid (17mg,0.055mmol) in N, N-dimethylformamide (0.5mL) were added 1- (4-chloro-3- (methylsulfonyl) phenyl) ethylamine (50mg,0.21mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (80mg,0.21mmol), and N, N-diisopropylethylamine (80 μ L,0.46 mmol). The reaction mixture was stirred at 50 ℃ for 16 hours. LC-MS showed the reaction was complete. The mixture was purified by preparative HPLC to give the final product as an off-white solid.
LC-MS:525.6[M+1]+；1H NMR(400MHz,DMSO-d6):9.18(d,1H,J＝7.3Hz),8.21(s,1H),8.10(d,1H,J＝1.8Hz),8.0-7.85(m,2H),7.80-7.65(m,4H),7.32(d,2H,J＝7.9Hz),5.30-5.20(m,1H),3.50(t,2H,J＝6.5Hz),3.46-3.35(m,5H),2.36(s,3H),2.00-1.78(m,4H),1.53(d,3H,J＝7Hz)。
Compound 49
4' -methyl-5- (pyrrolidine-1-carbonyl) -N- (quinolin-7-ylmethyl) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid (17mg,0.055mmol) in N, N-dimethylformamide (0.5mL) were added quinolin-7-ylmethylamine (34mg,0.22mmol) (WO2008/130481), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (80mg,0.21mmol) and N, N-diisopropylethylamine (80 μ L,0.46 mmol). The reaction mixture was stirred at 50 ℃ for 16 hours. LC-MS showed the reaction was complete. The reaction mixture was purified by preparative HPLC to give the title compound.
LC-MS:450.5[M+1]+；1H NMR(400MHz,CDCl3):8.92(dd,1H,J＝4.1,1.4Hz),8.19(d,1H,J＝8.1Hz),8.12(t,1H,J＝1.7Hz),8.09(s,1H),7.92-7.80(m,3H),7.60(dd,1H,J＝9.9,1.4Hz),7.52(d,2H,J＝8.1Hz),7.43(dd,1H,J＝8.3,4.3Hz),7.26(d,2H,J＝7.8Hz),6.76(t,1H,J＝5.7Hz),4.91(d,2H,J＝5.8Hz),3.66(t,2H,J＝6.9Hz),3.47(t,2H,J＝6.6Hz),2.40(s,3H),2.10-1.80(m,4H)。
Compound 58
4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (methylsulfonyl) biphenyl-3-carboxamide
To 4 '-methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (40mg,0.14mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (53mg,0.28mmol), 1-hydroxybenzotriazole hydrate (21mg,0.14mmol) and CH2Cl2(3mL) to the mixture was added (6-methylpyridin-3-yl) methylamine (25mg,0.21mmol) and N, N-diisopropylethylamine (48. mu.L, 0.28 mmol). The mixture was stirred at room temperature overnight and then concentrated. By preparative HPLC(100X20.2mm, C18 column; 30-80% CH3CN-Water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:395.5[M+1]+；1H NMR(400MHz,CDCl3):8.52(s,1H),8.34(s,1H),8.26(s,1H),8.18(s,1H),7.65(dd,1H,J＝8.0,2.0Hz),7.54(d,2H,J＝8.0Hz),7.30(d,2H,J＝8.0Hz),7.17(d,1H,J＝8.0Hz),6.73(bs,1H),4.66(d,2H,J＝5.6Hz),3.11(s,3H),2.56(s,3H),2.42(s,3H)。
Compound 62
2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) -N- ((6- (trifluoromethyl) pyridin-3-yl) methyl) biphenyl-3-carboxamide
A)2 '-cyano-4' -methylbiphenyl-3, 5-dicarboxylic acid dimethyl ester
To a Parr pressure reactor was added dimethyl 5-bromoisophthalate (1.3g,4.8mmol), 5-methyl-2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzonitrile (2.0g,5.9mmol), toluene (25mL), ethanol (5mL), cesium carbonate (1.7g,5.2mmol), and water (2.5 mL). The mixture was degassed, purged several times with nitrogen, and then tetrakis (triphenylphosphine) -palladium (0) (280mg,0.24mmol) was added. The tube was sealed and the mixture was heated at 90 ℃ overnight. After cooling, the mixture was diluted with EtOAc (200mL), washed with brine, and Na2SO4Drying and concentrating. The residue was purified by flash chromatography to afford the desired product as a white solid. LC-MS 310.5[ M +1 ]]+；1H NMR(400MHz,CDCl3):8.75(s,1H),8.39(d,J＝1.6Hz,2H),7.61(s,1H),7.49(dd,J＝1.1,8.5Hz,1H),7.43(d,J＝7.9Hz,1H),3.98(s,6H),2.46(s,3H)。
B)2 '-cyano-5- (methoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid
Adding 2 '-cyano-4' -methyl into a round-bottom flaskBiphenyl-3, 5-dicarboxylic acid dimethyl ester (1.5g,4.4mmol), ethanol (100mL), 1, 4-bisAlkane (20mL) and sodium hydroxide (0.17g,4.4mmol) in water (10 mL). The mixture was stirred at room temperature for 3 h. Volatiles were removed under reduced pressure and the residue was acidified to pH 4 with 1N aqueous HCl and CH2Cl2And (4) extracting. The combined organic layers were dried and concentrated. The residue was purified by flash chromatography to give the desired product 2 '-cyano-5- (methoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid as a white solid.1HNMR (400MHz, DMSO-d6):13.58(Br, 1H),8.56(t, J ═ 1.3Hz,1H),8.34-8.31(m,2H),7.85(s,1H),7.65(d, J ═ 2.6Hz,2H),3.92(s,3H),2.42(s, 3H). 2 '-cyano-4' -methylbiphenyl-3, 5-dicarboxylic acid 3-ethyl 5-methyl ester was also isolated as a white solid.
C)2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid methyl ester
To a mixture of 2' -cyano-5- (methoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid (0.20g,0.68mmol), N- (3-dimethylaminopropyl) -N ' -ethylcarbodiimide hydrochloride (0.26g,1.4mmol), 1-hydroxybenzotriazole hydrate (0.21g,1.4mmol) and dichloromethane (10mL) was added pyrrolidine (0.072g,1.0mmol) and N, N-diisopropylethylamine (0.24mL,1.4 mmol). The mixture was stirred at room temperature overnight and then concentrated. The residue was purified by flash chromatography (silica gel column, 0-100% EtOAc/hexanes) to give the desired product as a white solid. LC-MS 349.0[ M +1 ]]+；1H NMR(400MHz,DMSO-d6)8.14(s,1H),8.12(t,J＝1.6Hz,1H),7.96(t,J＝1.7Hz,1H),7.83(s,1H),7.66-7.60(m,2H),3.91(s,3H),3.52-3.43(m,4H),2.42(s,3H),1.90-1.81(m,4H)。
B)2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid
To a round-bottom flask was added methyl 2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylate (0.17g,0.46mmol), lithium hydroxide (17mg,0.70mmol), methanol (5mL) and water (1 mL). The mixture was stirred at room temperature overnight. Pressure reducing device Devolatilizing, acidifying the residue with 1N HCl, and adding CH2Cl2And (4) extracting. With Na2SO4The combined organic layers were dried, filtered, and concentrated to give the desired product as a white solid. LC-MS 335.4[ M +1 ]]+；1H NMR(400MHz,DMSO-d6):8.14(t,J＝1.6Hz,1H),8.10(t,J＝1.5Hz,1H),7.93(t,J＝1.7Hz,1H),7.83(t,J＝0.8Hz,1H),7.66-7.60(m,2H),3.51-3.42(m,4H),2.42(s,3H),1.91-1.83(m,4H)。
C)2 '-cyano-4' -methyl-5- (pyrrolidine-1-carbonyl) -N- ((6- (trifluoromethyl) pyridin-3-yl) methyl) biphenyl-3-carboxamide
To a round bottom flask was added 2' -cyano-4 ' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid (50mg,0.1mmol), N- (3-dimethylaminopropyl) -N ' -ethylcarbodiimide hydrochloride (52mg,0.27mmol), 1-hydroxybenzotriazole (36mg,0.27mmol), N-diisopropylethylamine (35mg,0.27mmol), dichloromethane (4mL), 4-dimethylaminopyridine (1mg) and C- (6-trifluoromethyl-pyridin-3-yl) -methylamine (47mg,0.27 mmol). The mixture was stirred at room temperature overnight and then with CH2Cl2Diluting with Na2HPO4Washing with aqueous solution, brine, and Na2SO4Drying and concentrating. The residue was purified by preparative HPLC to give the desired product as a white solid.
LC-MS:493.2[M+1]+；1H NMR(400MHz,CDCl3):8.67(s,1H),8.07(dd,J＝1.6Hz,6.0Hz,2H),7.87(dd,J＝1.1,8.4Hz,1H),7.72-7.71(m,1H),7.59-7.57(m,2H),7.47(dd,J＝0.44,8.1Hz,1H),7.41(d,J＝8Hz,1H),4.67(s,2H),3.54-3.49(m,4H),2.44(s,3H),1.95-1.88(m,4H)。
Compound 63
2 '-cyano-5- (3-hydroxyazetidine-1-carbonyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
A)2 '-cyano-5- (ethoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid
To a round-bottomed flask was added 2 '-cyano-4' -methylbiphenyl-3, 5-dicarboxylic acid 3-ethyl 5-methyl ester (0.12g,0.33mmol), 1, 4-bis Alkane (10mL) and lithium hydroxide (9mg,0.4mmol) in water (2 mL). The mixture was stirred at room temperature for 2 h. The volatiles were removed under reduced pressure, the residue was acidified with 1N HCl and CH2Cl2And (4) extracting. The separated organic phase was dried and concentrated. The residue was purified by flash chromatography to afford the desired product as a white solid.
B)2 '-cyano-5- (3-hydroxyazetidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
To a mixture of 2' -cyano-5- (ethoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid (90mg,0.29mmol), N- (3-dimethylaminopropyl) -N ' -ethylcarbodiimide hydrochloride (120mg,0.61mmol), 1-hydroxybenzotriazole hydrate (93mg,0.61mmol) and dichloromethane (5mL) was added 3-hydroxyazetidine hydrochloride (67mg,0.61mmol) and N, N-diisopropylethylamine (0.21mL,1.2 mmol). The mixture was stirred at room temperature overnight and then concentrated under reduced pressure. The residue was purified by flash chromatography to afford the desired product as a white solid. LC-MS 365.4[ M +1 ]]+；1H NMR(400MHz,DMSO-d6):8.24(d,J＝1.6Hz,1H),8.18(t,J＝1.7Hz,1H),8.00(t,J＝1.7Hz,1H),7.84(s,1H),7.64-7.61(m,2H),5.79(d,J＝6.0Hz,1H),4.54-4.52(m,2H),4.37(q,J＝7.2Hz,2H),4.35-4.34(m,1H),4.29-4.27(m,1H),3.84-3.82(m,1H),2.42(s,3H),1.35(t,J＝7.2Hz,3H)。
C)2 '-cyano-5- (3-hydroxyazetidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylic acid
To a round bottom flask was added ethyl 2 '-cyano-5- (3-hydroxyazetidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylate (25mg,0.066mmol), sodium hydroxide (10mg,0.25mmol), acetonitrile (2mL) and water (2 mL). Mixing the mixture Stir at room temperature overnight. 1N aqueous HCl (3mL) was added and the volatiles were removed under reduced pressure. By CH2Cl2Extracting the residue with CH2Cl2The layer was concentrated to dryness to give the desired product as a white solid. LC-MS 337.5[ M +1 ]]+。
D)2 '-cyano-5- (3-hydroxyazetidine-1-carbonyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
To a round bottom flask was added 2' -cyano-4 ' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (80mg,0.24mmol), N- (3-dimethylaminopropyl) -N ' -ethylcarbodiimide hydrochloride (80mg,0.42mmol), 1-hydroxybenzotriazole hydrate (64mg,0.42mmol), N-diisopropylethylamine (110mg,0.83mmol), dichloromethane (5mL) and 3-hydroxyazetidine hydrochloride (45mg,0.42 mmol). The mixture was stirred at room temperature overnight and then concentrated. The residue was purified by flash chromatography to afford the desired product as a white solid.
LC-MS:441.5[M+1]+；1H NMR(400MHz,CDCl3):8.54(s,1H),8.10(d,J＝8.3Hz,2H),7.81(s,1H),7.72(d,J＝7.9Hz,1H),7.58(s,1H),7.49-7.42(m,3H),7.18(d,J＝7,9Hz,1H),4.68-4.57(m,4H),4.44-4.41(m,1H),4.29-4.28(m,1H),4.05-4.02(m,1H),2.57(s,3H),2.45(s,3H)。
Compound 65
(R) -2,4' -dimethyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (pyrrolidin-1-ylsulfonyl) biphenyl-3-carboxamide
A)5- (chlorosulfonyl) -3-iodo-2-methylbenzoic acid
Chlorosulfonic acid (3.80mL,57.2mmol) was added to a round bottom flask and 3-iodo-2-methylbenzoic acid (5.00g,19.1mmol) was added in portions at 0 ℃. The reaction was heated at 95 ℃ for 2 hours and then stirred at room temperature overnight. The reaction mixture was poured onto ice and the solid formed was collected and dried to give the desired product as a white solid.
B) 3-iodo-2-methyl-5- (pyrrolidin-1-ylsulfonyl) benzoic acid
To a round bottom flask was added 5- (chlorosulfonyl) -3-iodo-2-methylbenzoic acid (1.00g,2.77mmol), pyrrolidine (0.278mL,3.33mmol), 1, 4-bisAlkane (4mL) and pyridine (0.20mL,2.5 mmol). The mixture was stirred for 30 minutes. The solvent was removed and the residue was purified by flash chromatography to give the compound as a yellow solid.
C)2,4' -dimethyl-5- (pyrrolidin-1-ylsulfonyl) biphenyl-3-carboxylic acid
To 3-iodo-2-methyl-5- (pyrrolidin-1-ylsulfonyl) benzoic acid (1.00g,2.53mmol), p-tolylboronic acid (0.378g,2.78mmol), toluene (10mL), ethanol (3mL), cesium carbonate (0.907g,2.78mmol), and water (1mL) was added tetrakis (triphenylphosphine) palladium (0) (146mg,0.126mmol) under an argon atmosphere. The mixture was heated to reflux for 6h, then cooled to room temperature and filtered through Celite. The filtrate was concentrated and the residue was purified by silica gel column (0-50% EtOAc/hexanes) to give a yellow solid.
D) (R) -2,4' -dimethyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (pyrrolidin-1-ylsulfonyl) biphenyl-3-carboxamide
To a reaction vial was added 2,4' -dimethyl-5- (pyrrolidin-1-ylsulfonyl) biphenyl-3-carboxylic acid (40mg,0.11mmol), (R) -1- (2-methylpyrimidin-5-yl) ethylamine (20mg,0.14mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (106mg,0.28mmol), N, N-diisopropylethylamine (39. mu.L, 0.22mmol), and N, N-dimethylformamide (1.33mL), and the reaction mixture was stirred at room temperature overnight. The reaction mixture was purified by preparative HPLC to afford the compound as a light brown solid.
LC-MS:479.4[M+1]+；1H NMR(400MHz,DMSO-d6):9.13(t,J＝7.34Hz,1H),8.73(s,2H),7.61(d,J＝1.87Hz,1H),7.54(d,J＝1.87Hz,1H),7.31(d,J＝7.68Hz,2H),7.25(d,J＝7.68Hz,2H),5.15(t,J＝7.45Hz,1H),3.16(t,J＝6.72Hz,4H),2.60(s,3H),2.37(s,3H),2.18(s,3H),1.70(t,J＝6.72Hz,4H),1.50(d,J＝6.72Hz,3H)。
Compound 67
2,4' -dimethyl-N- ((2-methylpyrimidin-5-yl) methyl) -5- (pyrrolidin-1-ylsulfonyl) biphenyl-3-carboxamide
To a reaction vial was added 2,4 '-dimethyl-5- (pyrrolidin-1-ylsulfonyl) biphenyl-3-carboxylic acid (40mg,0.11mmol), (2-methylpyrimidin-5-yl) methylamine (18mg,0.15mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (106mg,0.28mmol), N-diisopropylethylamine (39 μ L,0.22mmol), and N, N-dimethylformamide (1.33mL) and the reaction mixture was stirred overnight. The reaction mixture was purified by preparative HPLC to afford the compound as a white solid.
LC-MS:465.3[M+1]+；1H NMR(400MHz,DMSO-d6):9.14(t,J＝6.16Hz,1H),8.69(s,2H),7.65(d,J＝1.93Hz,1H),7.54(d,J＝1.93Hz,1H),7.30(d,J＝7.73Hz,2H),7.26(d,J＝7.73Hz,2H),4.46(d,J＝5.64Hz,2H),3.16(t,J＝6.78Hz,4H),2.60(s,3H),2.37(s,3H),2.22(s,3H),1.70(t,J＝6.67Hz,4H)。
Compound 73
N- (4-chloro-3- (N-cyclopropylsulfamoyl) benzyl) -4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-Carboxamides
To a solution of 4 '-methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid (60mg,0.19mmol) in N, N-dimethylformamide (1.0mL) were added 5- (aminomethyl) -2-chloro-N-cyclopropylbenzenesulfonamide (80mg,0.31mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (150mg,0.39mmol), and N, N-diisopropylethylamine (150 μ L,0.86 mmol). The reaction mixture was stirred at 50 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give a pale solid.
LC-MS:552.6[M+1]+；1H NMR(400MHz,DMSO-d6):9.39(t,1H,J＝5.9Hz),8.23(t,2H,J＝1.4Hz),8.00(d,1H,J＝1.6Hz),7.95(t,1H,J＝1.3Hz),7.92(t,1H,J＝1.5Hz),7.70-7.55(m,4H),7.32(d,2H,J＝ 8.0Hz),4.58(d,2H,J＝5.8Hz),3.50(t,2H,J＝6.5Hz),3.43(t,2H,J＝6.4Hz),2.36(s,3H),2.28-2.12(m,1H),2.00-1.78(m,4H),0.50-0.32(m,4H)。
Compound 83
N- (3-hydroxy-1- (6- (trifluoromethyl) pyridin-3-yl) propyl) -4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid (17mg,0.055mmol) in N, N-dimethylformamide (0.5mL) was added 3-amino-3- (6- (trifluoromethyl) pyridin-3-yl) propan-1-ol (47mg,0.21mmol) (WO2008/130481), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (80mg,0.21mmol) and N, N-diisopropylethylamine (80 μ L,0.46 mmol). The reaction mixture was stirred at 30 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give a white solid.
LC-MS:512.4[M+1]+；1H NMR(400MHz,DMSO-d6):9.15(d,1H,J＝7.6Hz),8.81(s,1H),8.19(s,1H),8.09(dd,1H,J＝8.0,1.3Hz),8.0-7.85(m,3H),7.66(d,2H,J＝8.1Hz),7.32(d,2H,J＝8.0Hz),5.4-5.25(m,1H),4.71(t,1H,J＝4.5Hz),3.60-3.35(m,6H),2.36(s,3H),2.25-2.10(m,1H),2.05-1.95(m,1H),1.94-1.78(m,4H)。
Compound 86
4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (3-morpholinopyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (55mg,0.15mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (58mg,0.30mmol), 1-hydroxybenzotriazole hydrate (23mg,0.15mmol) and CH2Cl2To a mixture (3mL) was added 4- (pyrrolidin-3-yl) morpholine (48mg,0.30mmol) and N, N-diisopropylethylamine (53. mu.L, 0.30 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-60% MeCN/water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:499.7[M+1]+；1H NMR(400MHz,CD3OD):8.43(d,1H,J＝2.0Hz),8.19(s,1H),7.92(d,2H,J＝1.6Hz),7.76(dd,1H,J＝8.0,2.0Hz),7.59(d,2H,J＝8.0Hz),7.31-7.27(m,3H),4.59(s,2H),3.95-3.20(m,8H),2.90(m,1H),2.65-2.10(m,11H),1.85(m,1H)。
Compound 87
4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid
A)4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid ethyl ester
To 5- (ethoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid (1.5g,5.3mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (2.0g,10mmol), 1-hydroxybenzotriazole hydrate (0.32g,2.1mmol) and CH2Cl2(50mL) to a mixture was added (6-methylpyridin-3-yl) methylamine (0.97g,7.9mmol) and N, N-diisopropylethylamine (1.8mL,10 mmol). The mixture was stirred at room temperature for 4h, then water and Na were used2CO3Washed with an aqueous solution and dried (Na)2SO4) And (4) concentrating in vacuum. The residue was purified by silica gel column to give a white foam.
LC-MS:389.4[M+1]+；1H NMR(400MHz,CDCl3):8.53(d,1H,J＝2.0Hz),8.39(t,1H,J＝1.6Hz),8.29(t,1H,J＝1.6Hz),8.26(t,1H,J＝1.6Hz),7.66(dd,1H,J＝8.0,2.0Hz),7.55(d,2H,J＝8.4Hz),7.28(d,2H,J＝8.0Hz),7.17(d,1H,J＝8.0Hz),6.64(m,1H),4.67(d,2H,J＝5.6Hz),4.42(q,2H,J＝7.2Hz),2.57(s,3H),2.41(s,3H),1.42(t,3H,J＝7.2Hz)。
B)4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid
A mixture of ethyl 4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylate (1.02g,2.62mmol), lithium hydroxide (310mg,13mmol), EtOH (100mL), and water (10mL) was stirred at rt overnight. LC-MS showed the reaction was complete. The solvent was removed in vacuo and the residue treated with water (50mL) and acidified to pH 4 with 1N aqueous HCl. The precipitated solid was collected by filtration, washed with water, and dried to give a white solid.
LC-MS:361.3[M+1]+；1H NMR(400MHz,DMSO-d6):13.5(bs,1H),9.34(t,1H,J＝5.6Hz),8.44(d,1H,J＝2.0Hz),8.41(t,1H,J＝1.6Hz),8.34(t,1H,J＝1.6Hz),8.29(t,1H,J＝1.6Hz),7.67(d,2H,J＝8.4Hz),7.64(dd,1H,J＝8.0,2.0Hz),7.33(d,2H,J＝8.0Hz),7.22(d,1H,J＝8.0Hz),4.49(d,2H,J＝5.6Hz),2.44(s,3H),2.37(s,3H)。
Compound 94
(R) -5- (3-hydroxypyrrolidine-1-carbonyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
A) (R) -5- (3-hydroxypyrrolidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
To 5- (ethoxycarbonyl) -4 '-methylbiphenyl-3-carboxylic acid (1.50g,5.28mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (2.0g,10mmol), 1-hydroxybenzotriazole hydrate (0.404g,2.64mmol) and CH2Cl2(30mL) to the mixture was added (R) -3-hydroxypyrrolidine (0.92g,10mmol) and N, N-diisopropylethylamine (1.8mL,10 mmol). The mixture was stirred at room temperature overnight and then with CH2Cl2(100mL) diluted with NaHCO3Washed with aqueous solution, brine and dried (Na)2SO4) And (4) concentrating in vacuum. The residue was purified by silica gel column (100% EtOAc) to give a white foam. LC-MS 354.2[ M +1 ]]+；1H NMR(400MHz,CDCl3) 8.32(s,1H),8.16 and 8.12(bs,1H),7.92(m,1H),7.53(d,2H, J ═ 7.6Hz),7.28(d,2H, J ═ 8.4Hz),4.63 and 4.49(bs,1H),4.41(q,2H, J ═ 7.2Hz),3.95-3.40(m,4H),2.41(s,3H),2.20-1.90(m,2H),1.41(t,3H, J ═ 7.2 Hz).
B) (R) -5- (3-hydroxypyrrolidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylic acid
A mixture of (R) -ethyl 5- (3-hydroxypyrrolidine-1-carbonyl) -4' -methylbiphenyl-3-carboxylate (1.25g,3.54mmol), lithium hydroxide (0.42g,18mmol), methanol (50mL) and water (5mL) was stirred at rt for 4 h. LC-MS showed the reaction was complete. Volatiles were removed in vacuo and the residue was treated with water, acidified to pH2-3 with 1N aqueous HCl and extracted with EtOAc (50mL x 3). The combined organic layers were washed with brine and dried (Na)2SO4) And concentrated to give a white foam. LC-MS 326.3[ M +1 ]]+；1H NMR(400MHz,CD3OD) 8.33(m,1H),8.11 and 8.10(t,1H, J-1.6 Hz),7.98 and 7.97(t,1H, J-1.6 Hz),7.57(d,2H, J-7.6 Hz),7.30(d,2H, J-8.0 Hz),4.51 and 4.38(m,1H),3.85-3.35(m,4H),2.39(s,3H),2.20-1.90(m, 2H).
C) (R) -5- (3-hydroxypyrrolidine-1-carbonyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
To 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (55mg,0.15mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (58mg,0.30mmol), 1-hydroxybenzotriazole hydrate (23mg,0.15mmol) and CH2Cl2(3mL) to the mixture was added (R) -3-hydroxypyrrolidine (26mg,0.30mmol) and N, N-diisopropylethylamine (53. mu.L, 0.30 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-60% MeCN/water [10mM Et2NH]) Then column on silica gel (0-20% MeOH/CH)2Cl2) The residue was purified to give a white foam.
LC-MS:430.3[M+1]+；1H NMR(400MHz,CD3OD) 8.44(d,1H, J ═ 2.0Hz),8.19(m,1H),7.95-7.90(m,2H),7.77(dd,1H, J ═ 8.0,2.4Hz),7.59(d,2H, J ═ 8.0Hz),7.29(d,3H, J ═ 8.0Hz),4.59(s,2H),4.50 and 4.37(m,1H),3.85-3.30(m,4H),2.51(s,3H),2.38(s,3H),2.20-1.90(m, 2H).
Compound 95
(S) -5- (3-hydroxypyrrolidine-1-carbonyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
To 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (55 mg,0.15mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (58mg,0.30mmol), 1-hydroxybenzotriazole hydrate (23mg,0.15mmol) and CH2Cl2(3mL) to the mixture was added (S) -3-hydroxypyrrolidine (26mg,0.30mmol) and N, N-diisopropylethylamine (53. mu.L, 0.30 mmol). The mixture was stirred at room temperature overnight and then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, 30-60% MeCN/water [10mM Et2NH]) Then column on silica gel (0-20% MeOH/CH)2Cl2) The residue was purified to give a white foam.
LC-MS:430.3[M+1]+；1H NMR(400MHz,CD3OD) 8.44(d,1H, J ═ 2.0Hz),8.19(m,1H),7.95-7.90(m,2H),7.77(dd,1H, J ═ 8.0,2.4Hz),7.59(d,2H, J ═ 8.0Hz),7.29(d,3H, J ═ 8.0Hz),4.59(s,2H),4.50 and 4.37(m,1H),3.81-3.30(m,4H),2.51(s,3H),2.38(s,3H),2.20-1.90(m, 2H).
Compound 108
(R) -N- (1- (5-chloro-1-methyl-1H-pyrazol-4-yl) ethyl) -4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To a 20mL vial was added 4 '-methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid (30mg,0.097mmol), (R) -1- (5-chloro-1-methyl-1H-pyrazol-4-yl) ethylamine hydrochloride (24mg,0.12mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (92mg,0.24mmol), N-diisopropylethylamine (0.068mL,0.39mmol), and N, N-dimethylformamide (1.0mL), and the reaction mixture was stirred at room temperature overnight. The reaction mixture was purified by preparative HPLC to give the product as a light yellow solid.
LC-MS:451.3[M+1]+；1H NMR(DMSO-d6):8.87(d,J＝7.77Hz,1H),8.16(t,J＝1.83Hz,1H),7.92(t,J＝1.61Hz,1H),7.87(t,J＝1.61Hz,1H),7.66(d,J＝8.22Hz,2H),7.59(s,1H),7.31(d,J＝8.22Hz,2H),5.16-5.12(m,1H),3.76(s,3H),3.49(t,J＝6.70Hz,2H),3.40(t,J＝6.70Hz,2H),2.36(s,3H),1.90-1.80(m,4H),1.47(d,J＝7.45Hz,3H)。
Compound 111
4' -methyl-N- (1- (2-methylthiazol-4-yl) ethyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To a 20mL vial was added 4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid (30mg,0.097mmol), 1- (2-methylthiazol-4-yl) -ethylamine hydrochloride (22mg,0.12mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (92mg,0.24mmol), N, N-diisopropylethylamine (0.068mL,0.39mmol), and N, N-dimethylformamide (1.0mL), and the reaction mixture was stirred at room temperature overnight. The reaction mixture was purified by preparative HPLC to give the product as a light peach colored solid.
LC-MS:434.4[M+1]+；1H NMR(DMSO-d6):9.01(d,J＝8.50Hz,1H),8.24(t,J＝1.59Hz,1H),7.97(t,J＝1.99Hz,1H),7.88(t,J＝1.99Hz,1H),7.68(d,J＝8.35Hz,2H),7.31(d,J＝7.95Hz,2H),7.25(d,J＝0.99Hz,1H),5.31-5.27(m,1H),3.50(t,J＝6.53Hz,2H),3.42(t,J＝6.53Hz,2H),2.63(s,3H),2.36(s,3H),1.91-1.81(m,4H),1.53(d,J＝7.26Hz,3H)。
Compound 113
3- (5-methylpyridin-2-yl) -N- ((6-methylpyridin-3-yl) methyl) -5- (methylsulfonyl) benzamide
A) 3-bromo-5- (methylsulfonyl) benzoic acid
A mixture of methyl 3-bromo-5- (methylsulfonyl) benzoate (0.65g,2.2mmol), lithium hydroxide (0.26g,11mmol), tetrahydrofuran (25mL) and water (5mL) was addedStirred at room temperature for 3 h. Water (50mL) was added and the mixture was acidified to pH2-3 with 1N aqueous HCl and extracted with EtOAc (100 mL). The organic layer was separated, washed with brine and dried (Na)2SO4) And concentrated to give a white solid. LC-MS 278.6[ M-1 ]]-；1H NMR(400MHz,CDCl3): 8.59(m,1H),8.50(m,1H),8.33(m,1H),3.13(s,3H)。
B) 3-bromo-N- ((6-methylpyridin-3-yl) methyl) -5- (methylsulfonyl) benzamide
To 3-bromo-5- (methylsulfonyl) benzoic acid (630mg,2.2mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (860mg,4.5mmol), 1-hydroxybenzotriazole hydrate (140mg,0.90mmol) and CH2Cl2(25mL) to the mixture was added (6-methylpyridin-3-yl) methylamine (550mg,4.5mmol) and N, N-diisopropylethylamine (0.79mL,4.5 mmol). The mixture was stirred at room temperature overnight and then with CH2Cl2(50mL) dilution. With water and Na2CO3The organic phase is washed with an aqueous solution and dried (Na)2SO4) And (4) concentrating in vacuum. The residue was purified by silica gel column (50-100% EtOAc/hexanes) to give a white solid. LC-MS 385.2[ M +1 ]]+；1H NMR(400MHz,CDCl3):8.51(d,1H,J＝1.6Hz),8.28(t,1H,J＝1.6Hz),8.23(t,1H,J＝1.6Hz),8.19(t,1H,J＝1.6Hz),7.66(dd,1H,J＝8.0,2.0Hz),7.19(d,1H,J＝8.0Hz),6.93(bs,1H),4.63(d,2H,J＝5.6Hz),3.09(s,3H),2.57(s,3H)。
C)3- (5-methylpyridin-2-yl) -N- ((6-methylpyridin-3-yl) methyl) -5- (methylsulfonyl) benzamide
A mixture of 3-bromo-N- ((6-methylpyridin-3-yl) methyl) -5- (methylsulfonyl) benzamide (71mg,0.18mmol), 5-methyl-2- (tributylstannyl) pyridine (91mg,0.23mmol), tetrakis (triphenylphosphine) palladium (0) (11mg,0.0093mmol), and toluene (2.0mL) was subjected to microwave irradiation at 120 ℃ for 2 hours under an argon atmosphere. The mixture was cooled to precipitate the product. The solvent was discarded and the precipitated solid was washed with hexane and then subjected to preparative HPLC (100X21.2mm C18 column, 30-70% CH)3CN/Water [10mM Et2NH]) Purification gave a white foam.
LC-MS:396.4[M+1]+；1H NMR(400MHz,CDCl3):8.72(t,1H,J＝1.6Hz),8.69(t,1H,J＝1.6Hz),8.53(m,2H),8.34(t,1H,J＝1.6Hz),7.76(d,1H,J＝8.0Hz),7.68-7.62(m,2H),7.17(d,1H,J＝8.0Hz),6.86(bs,1H),4.67(d,2H,J＝6.4Hz),3.13(s,3H),2.57(s,3H),2.41(s,3H)。
Compound 116
(R) -4' -methyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (methylsulfonyl) biphenyl-3-carboxamide
To a solution of 4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (1.0g,3.44mmol) in N, N-dimethylformamide (10mL) were added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (900mg,5.58mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (3.0g,7.89mmol), and N, N-diisopropylethylamine (3.0mL,17.22 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. LC-MS showed the reaction was complete. The reaction solution was diluted with EtOAc, and saturated NaHCO with water3Washing with aqueous solution, brine, anhydrous MgSO4And (5) drying. The residue was purified by silica gel column and preparative HPLC to give the final product as a pale solid.
LC-MS:410.1[M+1]+；1H NMR(400MHz,DMSO-d6):9.26(d,1H,J＝7.4Hz),8.75(s,2H),8.44(t,1H,J＝1.6Hz),8.33(t,1H,J＝1.6Hz),8.28(t,1H,J＝1.6Hz),7.75(d,2H,J＝8.2Hz),7.37(d,2H,J＝7.9Hz),5.22(m,1H),3.34(s,3H),2.60(s,3H),2.38(s,3H),1.58(d,3H,J＝7.1Hz)。
Compound 122
3- (5-methylpyridin-2-yl) -N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) benzamide
A)3- (5-methylpyridin-2-yl) -5- (pyrrolidine-1-carbonyl) benzoic acid methyl ester
A mixture of methyl 3-bromo-5- (pyrrolidine-1-carbonyl) benzoate (1.2g,3.8mmol), 5-methyl-2- (tributylstannyl) pyridine (0.90mL,2.6mmol), tetrakis (triphenylphosphine) palladium (0) (100mg,0.086mmol) was subjected to microwave irradiation at 120 ℃ for 2 hours under an argon atmosphere. The mixture was cooled to room temperature and the precipitate was collected by filtration and washed with hexanes to give the title compound as a white solid. The filtrate was concentrated and purified by flash chromatography to give a further batch of the desired product. LC-MS 325.1[ M +1 ] ]+。
B)3- (5-methylpyridin-2-yl) -5- (pyrrolidine-1-carbonyl) benzoic acid
A round-bottom flask was charged with methyl 3- (5-methylpyridin-2-yl) -5- (pyrrolidine-1-carbonyl) benzoate (0.80g,2.47mmol), methanol (40mL), sodium hydroxide (0.20g,5.0mmol), and water (10 mL). The mixture was stirred at room temperature for 1 h. The volatiles were removed under reduced pressure. The residue was treated with 1n hcl (10mL), concentrated, and purified by preparative HPLC to give the desired product. LC-MS 311.5[ M +1 ]]+；1H NMR(400MHz,DMSO-d6):8.61(t,J＝1.7Hz,1H),8.52(t,J＝0.8Hz,1H),8.16(t,J＝1.8Hz,1H),7.99(t,J＝1.5Hz,1H),7.80(d,J＝8.2Hz,1H),7.72-7.69(m,1H),3.50(t,J＝6.7Hz,2H),3.41(t,J＝6.5Hz,2H),2.35(s,3H),1.91-1.81(m,4H)。
C)3- (5-methylpyridin-2-yl) -N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) benzamide
In a round bottom flask was added 3- (5-methylpyridin-2-yl) -5- (pyrrolidine-1-carbonyl) benzoic acid (80mg,0.26mmol), (6-methylpyridin-3-yl) methylamine (60mg,0.49mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (90mg,0.47mmol), 1-hydroxybenzotriazole hydrate (80mg,0.52mmol), N-diisopropylethylamine (80mg,0.62mmol) and dichloromethane (5 mL). The mixture was stirred at room temperature overnight and then concentrated. The residue was purified by preparative HPLC to give the desired product as a white solid.
LC-MS:415.5[M+1]+；1H NMR(400MHz,CD3OD):8.56(t,J＝1.7Hz,1H),8.53-8.52(m,1H),8.46(d,J＝2.1Hz,1H),8.29(t,J＝1.6Hz,1H),8.04(t,J＝1.6Hz,1H),7.89(d,J＝8.1Hz,1H),7.81-7.77(m,2H),7.32(d,J＝8.0Hz,1H),4.63(s,2H),3.66(t,J＝7.0Hz,2H),3.54(t,J＝6.7Hz,2H),2.54(s,3H),2.43(s,3H),2.05-1.94(m,4H)。
Compound 127
4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (4- (thiazol-2-yl) piperazine-1-carbonyl) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (30 mg,0.083mmol) in N, N-dimethylformamide (1mL) were added 2- (piperazin-1-yl) thiazole (40mg,0.24mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (80mg,0.21mol), and N, N-diisopropylethylamine (100 μ L,0.57 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give the final product as a white solid.
LC-MS:512.4[M+1]+；1H NMR(400MHz,DMSO-d6):9.27(t,1H,J＝5.6Hz),8.44(bs,1H),8.24(bs,1H),7.90-7.84(m,2H),7.70-7.60(m,3H),7.32(d,2H,J＝7.9Hz),7.22(d,1H,J＝8.0Hz),7.19(d,1H,J＝3.6Hz),6.89(d,1H,J＝3.6Hz),4.49(d,2H,J＝5.6Hz),3.90-3.70(m,2H),3.60-3.40(m,6H),2.44(s,3H),2.36(s,3H)。
Compound 130
5- (3-hydroxyazetidine-1-carbonyl) -4' -methyl-N- ((6- (trifluoromethyl) pyridin-3-yl) methyl) biphenyl-3-carboxamide
To a solution of 5- (3-hydroxyazetidine-1-carbonyl) -4 '-methylbiphenyl-3-carboxylic acid (80mg,0.26mmol) in N, N-dimethylformamide (1.5mL) were added C- (6-trifluoromethyl-pyridin-3-yl) -methylamine (70mg,0.40mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (200mg,0.53mmol) and N, N-diisopropylethylamine (200 μ L,1.15 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give a white solid.
LC-MS:470.5[M+1]+；1H NMR(400MHz,DMSO-d6):9.42(t,1H,J＝5.7Hz),8.78(d,1H,J＝1.3Hz),8.28(t,1H,J＝1.6Hz),8.06(t,1H,J＝1.4Hz),8.04(dd,1H,J＝8.0,1.5Hz),7.96(t,1H,J＝1.5Hz),7.89(d,1H,J＝8.1Hz),7.67(d,2H,J＝8.2Hz),7.33(d,2H,J＝8.0Hz),5.79(d,1H,J＝5.7Hz),4.65(d,2H,J＝5.7Hz),4.60-4.45(m,2H),4.28(m,1H),4.09(m,1H),3.82(m,1H),2.36(s,3H)。
Compound 140
4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- ((3aR,6aS) -octahydropyrrolo [3,4-c ] pyrrole-2-carbonyl) biphenyl-3-carboxamide
To a solution of 4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (25mg,0.069mmol) in N, N-dimethylformamide (1mL) was added (3aR,6aS) -hexahydropyrrolo [3,4-c ] -]Pyrrole-2 (1H) -carboxylic acid tert-butyl ester (30mg,0.14mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (60mg,0.16mmol) and N, N-diisopropylethylamine (100 μ L,0.57 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to afford the Boc-protected product as a white solid, which was dissolved in dichloromethane (3.0mL) and trifluoroacetic acid (0.20mL,2.6mmol) was added. The reaction mixture was stirred at rt overnight. Volatiles were removed in vacuo and subjected to preparative HPLC (100X21.2mm C18 column, 20-60% MeCN/water [10mM Et2NH]) The residue was purified to give the product as a white foam.
LC-MS:455.4[M+1]+；1H NMR(400MHz,DMSO-d6):9.26(t,1H,J＝5.3Hz),8.44(bs,1H),8.21(bs,1H),7.95-7.80(m,2H),7.70-7.60(m,3H),7.31(d,2H,J＝7.9Hz),7.22(d,1H,J＝7.9Hz),4.48(d,1H,J＝5.5Hz),3.90-3.15(m,4H),2.90(m,1H),2.85-2.60(m,4H),2.50-2.30(m,2H),2.44(s,3H),2.36(s,3H)。
Compound 162
N3,4' -dimethyl-N5- ((6-methylpyridin-3-yl) methyl) -N3- (pyridin-4-ylmethyl) biphenyl-3, 5-dicarboxamide
To a solution of 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (60mg,0.16mmol) in N, N-dimethylformamide (1mL) were added methyl-pyridin-4-ylmethyl-amine (50mg,0.41mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (140mg,0.37mmol) and N, N-diisopropylethylamine (140 μ L,0.80 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to afford the product as a light brown solid.
LC-MS:465.5[M+1]+；1H NMR (400MHz, DMSO-d6) (rotomers) 9.29(m,1H),8.57(m,2H),8.44(bs,1H),8.20(m,1H),8.95-7.55(m,5H),7.48-7.15(m,5H),4.74(bs,1H),4.60-4.40(m,3H),3.00 and 2.94(s,3H),2.44(s,3H),2.37 and 2.33(s, 3H).
Compound 163
N3,4' -dimethyl-N5- ((6-methylpyridin-3-yl) methyl) -N3- (pyridin-4-yl) biphenyl-3, 5-dicarboxamide
To a solution of 4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (50mg,0.134mmol) in N, N-dimethylformamide (1mL) were added N-methylpyridin-4-amine (40mg,0.37mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (120. mu.L, 0.69 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give the final product as a pale solid.
LC-MS:451.6[M+1]+；1H NMR(400MHz,DMSO-d6):9.20(t,1H,J＝5.7Hz),8.48-8.38(m,3H),8.12(t,1H,J＝1.6Hz),7.87(t,1H,J＝1.4Hz),7.65(t,1H,J＝1.5Hz),7.59(dd,1H,J＝8.0,2.3Hz),7.42(d,2H,J＝8.1Hz),7.30-7.15(m,5H),4.45(d,2H,J＝5.7Hz),3.45(s,3H),2.44(s,3H),2.33(s,3H)。
Compound 165
(R) -N3,4' -dimethyl-N5- ((6-methylpyridin-3-yl) methyl) -N3- (pyrrolidin-3-yl) biphenyl-3, 5-dicarboxamide
To a solution of 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (50mg,0.14mmol) in N, N-dimethylformamide (1mL) were added (R) -N-methylpyrrolidin-3-amine (50mg,0.50mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (120 μ L,0.69 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction solution was purified by preparative HPLC to give the final product as a pale solid.
LC-MS:443.5[M+1]+；1H NMR (400MHz, DMSO-d6):9.26(t,1H, J ═ 5.8Hz),8.43(bs,1H),8.22(bs,1H),7.94(bs,1H),7.88(bs,1H),7.70-7.60(m,3H),7.31(d,2H J ═ 7.9Hz),7.21(d,1H, J ═ 8.0Hz),4.48(d,2H, J ═ 5.7Hz),3.65-3.05(m,5H),2.44(s,3H),2.36(s,3H),2.29 and 2.17 (rotamers: s,3H),2.05-1.65(m, 3H).
Compound 166
N3, N3,4' -trimethyl-N5- ((6-methylpyridin-3-yl) methyl) biphenyl-3, 5-dicarboxamide
To a solution of 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (50mg,0.14mmol) in N, N-dimethylformamide (1mL) were added dimethylamine hydrochloride (60mg,0.74mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (0.50mL,2.87 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction solution was purified by preparative HPLC to give the final product as a light brown solid.
LC-MS:388.5[M+1]+；1H NMR(400MHz,DMSO-d6):9.26(t,1H,J＝5.8Hz),8.44(bs,1H),8.21(bs,1H),7.90-7.60(m,5H),7.31(d,2H,J＝7.3Hz),7.21(d,1H,J＝8.0Hz),4.49(d,2H,J＝4.9Hz),3.02(s,3H),2.94(s,3H),2.44(s,3H),2.36(s,3H)。
Compound 168
N3,4' -dimethyl-N5- ((6-methylpyridin-3-yl) methyl) -N3- (2,2, 2-trifluoroethyl) biphenyl-3, 5-dicarboxamide
To a solution of 4 '-methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (50mg,0.14mmol) in N, N-dimethylformamide (1.5mL) were added 2,2, 2-trifluoro-N-methylethylamine hydrochloride (60mg,0.40mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (0.50mL,2.87 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give the final product as a light yellow solid.
LC-MS:456.3[M+1]+；1H NMR(400MHz,DMSO-d6):9.31(bs,1H),8.44(bs,1H),8.25(bs,1H),7.90-7.60(m,5H),7.32(d,2H,J＝8.1Hz),7.21(d,1H,J＝8.0Hz),4.49(d,2H,J＝5.7Hz),4.45-4.10(m,2H),3.07(bs,3H),2.44(s,3H),2.36(s,3H)。
Compound 181
(R) -3- (5-methylpyridin-2-yl) -5- (methylsulfonyl) -N- (1- (6- (trifluoromethyl) pyridin-3-yl) ethyl) benzamide
A)3- (5-methylpyridin-2-yl) -5- (methylsulfonyl) benzoic acid methyl ester
A mixture of methyl 3-bromo-5- (methylsulfonyl) benzoate (1.0g,3.4mmol), 5-methyl-2- (tributylstannyl) pyridine (1.2mL,3.5mmol), tetrakis (triphenylphosphine) palladium (0) (200mg,0.17mmol), and toluene (10mL) was subjected to microwave irradiation at 120 ℃ for 2 hours under an argon atmosphere. The mixture was cooled to room temperature. The precipitate was collected by filtration and washed with hexane to give the title compound as a light brown solid (432 mg). The filtrate was concentrated and the residue was purified by flash chromatography to give a further crop of product as a pale yellow solid (200 mg).1HNMR(400MHz,CDCl3):8.91(t,J＝1.4Hz,1H),8.80(d,J＝1.7Hz, 1H),8.61(s,1H),8.58(d,J＝0.8Hz,1H),7.76(d,J＝8.1Hz,1H),7.65(d,J＝8.1Hz,1H),4.00(s,3H),3.15(s,3H),2.42(s,3H)。
B)3- (5-methylpyridin-2-yl) -5- (methylsulfonyl) benzoic acid (HCl salt)
To a round bottom flask was added methyl 3- (5-methylpyridin-2-yl) -5- (methylsulfonyl) benzoate (0.60g,1.96mmol), barium hydroxide octahydrate (1.23g,3.90mmol) and methanol (50 mL). The mixture was stirred at room temperature until the reaction was complete. The mixture was acidified with HCl (2M in ether, 20 mL). Concentrating the mixture to obtain the desired product and BaCl2As a white solid (1.4g, 40% purity). LC-MS 291.5[ M +1 ] ]；1H NMR(400MHz,DMSO-d6):8.90(s,1H),8.80(d,J＝1.7Hz,1H),8.63(s,1H),8.43(d,J＝1.5Hz,1H),8.16(d,J＝8.1Hz,1H),7.89(d,J＝7.8Hz,1H),7.64-7.55(m,1H),3.38(s,3H),2.40(s,3H)。
C) (R) -3- (5-methylpyridin-2-yl) -5- (methylsulfonyl) -N- (1- (6- (trifluoromethyl) pyridin-3-yl) ethyl) benzamide
To a round bottom flask was added 3- (5-methylpyridin-2-yl) -5- (methylsulfonyl) benzoic acid (40% BaCl2250mg,0.34mmol), (R) -1- (6- (trifluoromethyl) pyridin-3-yl) ethylamine (130mg,0.69mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (130mg,0.69mmol), 1-hydroxybenzotriazole hydrate (100mg,0.69mmol), triethylamine (87mg,0.86mmol) and CH2Cl2(3 mL). The mixture was stirred at room temperature overnight and then concentrated. The residue was purified by flash chromatography to afford the desired product as a white solid.
LC-MS:464.8[M+1]+；1H NMR(400MHz,DMSO-d6):9.41(d,J＝7.3Hz,1H),8.86(t,J＝1.6Hz,2H),8.74(t,J＝1.6Hz,1H),8.60(d,J＝2.2Hz,1H),8.43(t,J＝1.6Hz,1H),8.13-8.10(m,2H),7.91(d,J＝8.1Hz,1H),7.82(dd,J＝2.2,8.7Hz,1H),5.34(p,J＝7.1Hz,1H),3.33(s,3H),2.38(s,3H),1.60(d,J＝7.1Hz,3H)。
Compound 185
5- (isoindoline-2-carbonyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
To a solution of 4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) biphenyl-3-carboxylic acid (50mg,0.14mmol) in N, N-dimethylformamide (1mL) were added isoindoline (40mg,0.34mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (120. mu.L, 0.69 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction solution was purified by preparative HPLC to give the final product as a yellow solid.
LC-MS:461.9[M+1]+；1H NMR(400MHz,DMSO-d6):9.27(t,1H,J＝5.7Hz),8.45(d,1H,J＝2.0Hz),8.26(t,1H,J＝1.5Hz),8.02(bs,2H),7.72-7.62(m,3H),7.42(d,1H,J＝7.2Hz),7.35-7.25(m,5H),7.22(d,1H,J＝8.0Hz),4.90(s,2H),4.81(s,2H),4.49(d,2H,J＝5.7Hz),2.44(s,3H),2.36(s,3H)。
Compound 187
4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (1,2,3, 4-tetrahydroisoquinoline-2-carbonyl) biphenyl-3-carboxamide
To a solution of 4' -methyl-5- ((6-methylpyridin-3-yl) methylcarbamoyl) -biphenyl-3-carboxylic acid (50mg,0.14mmol) in N, N-dimethylformamide (1mL) were added 1,2,3, 4-tetrahydroisoquinoline (50mg,0.38mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (120. mu.L, 0.69 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction solution was purified by preparative HPLC to give the final product as a yellow solid.
LC-MS:476[M+1]+；1H NMR (400MHz, DMSO-d6):9.27(t,1H, J ═ 5.6Hz),8.44(d,1H, J ═ 1.8Hz),8.25(bs,1H),7.88(m,2H),7.72-7.62(m,3H),7.35-7.00(m,7H),4.81 and 4.61(bs,2H),4.48(d,2H, J ═ 5.7Hz),3.88 and 3.60(bs,2H),2.85(m,2H),2.44(s,3H),2.36(s, 3H).
Compound 191
4' -methyl-N- ((R) -1- (2-methylpyrimidin-5-yl) ethyl) -5- (octahydropyrrolo [1,2-a ] pyrazine-2-carbonyl) biphenyl-3-carboxamide
A)4' -methyl-5- (octahydropyrrolo [1,2-a ] pyrazine-2-carbonyl) biphenyl-3-carboxylic acid
To a solution of 5- (ethoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid (300mg,1.06mmol) in N, N-dimethylformamide (6mL) was added octahydropyrrolo [1,2-a ] ]Pyrazine (200mg,1.58mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (1.0g,2.63mmol) and N, N-diisopropylethylamine (1.0mL,5.74 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. LC-MS showed the reaction was complete. The reaction mixture was diluted with EtOAc, and saturated NaHCO with water3Washing with aqueous solution, brine, anhydrous MgSO4Drying and concentrating. The residue was purified by silica gel column to give the ethyl ester product as a brown foam. A mixture of the resulting ethyl ester (480mg,1.22mmol), lithium hydroxide (75mg,3.1mmol), MeOH (25mL), and water (4mL) was stirred at rt for 20 hr. LC-MS showed the reaction was complete. Volatiles were removed in vacuo and the residue was treated with water and acidified to pH 5 with 1N aqueous HCl. The aqueous phase was extracted with EtOAc (3 × 100mL), and the combined organic layers were washed with brine, dried, and concentrated to give the acid product as a pale solid.
B)4' -methyl-N- ((R) -1- (2-methylpyrimidin-5-yl) ethyl) -5- (octahydropyrrolo [1,2-a ] pyrazine-2-carbonyl) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (octahydropyrrolo [1,2-a ] pyrazine-2-carbonyl) biphenyl-3-carboxylic acid (50mg,0.14mmol) in N, N-dimethylformamide (1.0mL) were added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (35mg,0.25mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (120 μ L,0.69 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give the final product as a light yellow solid.
LC-MS:484.1[M+1]+；1H NMR (400MHz, DMSO-d6):9.07(d, 1H, J ═ 7.5Hz),8.73(s,2H),8.19(bs,1H),7.82(bs,1H),7.77(bs,1H),7.66(d,2H, J ═ 7.8Hz),7.32(d,2H, J ═ 8.0Hz),5.20(m,1H),4.63 and 4.49(bs,1H),3.70-2.80(m,4H),2.64(s,3H),2.36(s,3H),2.20-1.10(m,8H),1.55(d,3H, J ═ 7.1 Hz).
Compound 200
4' -bromo-N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
A)4' -bromo-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylic acid ethyl ester
To ethyl 3-bromo-5- (pyrrolidine-1-carbonyl) benzoate (3.20g,9.81mmol), 4-bromophenylboronic acid (2.17g,10.8mmol), toluene (40mL), cesium carbonate (3.52g,10.8mmol), and water (5mL) was added tetrakis (triphenylphosphine) palladium (0) (567mg,0.49mmol) under an argon atmosphere. The mixture was heated under reflux for 15 h. After cooling, the mixture was filtered through Celite and the filter cake was washed with EtOAc. The filtrate was washed with brine and dried (Na)2SO4) And (4) concentrating. The residue was purified by silica gel column (0-80% EtOAc/hexanes) to give a syrup. LC-MS 404.0[ M +1 ]]+。
B)4' -bromo-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid
A mixture of ethyl 4' -bromo-5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxylate (3.3g,8.2mmol), MeOH (100mL), water (20mL), and lithium hydroxide (0.90g,38mmol) was stirred at room temperature for 3 h. The mixture was concentrated in vacuo and the residue was acidified to pH2-3 with 1N aqueous HCl and extracted with EtOAc (2x100 mL). The combined organic layers were dried (Na)2SO4) And concentrated to give a white foam. LC-MS 376.1[ M +1 ]]+。
C)4' -bromo-N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
To 4 '-bromo-5- (pyrrolidine-1-carbonyl) -biphenyl-3-carboxylic acid (1.65g,4.41mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (1.7g,8.8mmol), 1-hydroxybenzotriazole hydrate (0.20g,1.3mmol) and CH2Cl2(50mL) of the mixture was added (6-methylpyridin-3-yl) methylamine (0.81g,6.6mmol) and N, N-diisopropylethylamine (1.5mL,8.8 mmol). The mixture was stirred at room temperature overnight, then washed with water and dried (Na)2SO4) And (4) concentrating. Passing through a silica gel column (0-15% MeOH/CH)2Cl2) The residue was purified by preparative HPLC (100 × 20.2mm, C18 column; 30-80% CH3CN-Water [10mM ET2NH]) The analytical sample was purified to give a white foam.
LC-MS:480.1[M+1]+；1H NMR(400MHz,DMSO-d6):9.26(t,1H,J＝5.6Hz),8.44(d,1H,J＝2.0Hz),8.23(t,1H,J＝1.6Hz),7.99(t,1H,J＝1.6Hz),7.94(t,1H,J＝1.6Hz),7.77-7.66(m,4H),7.63(dd,1H,J＝8.0,2.0Hz),7.22(d,1H,J＝8.0Hz),4.49(d,2H,J＝5.6Hz),3.50(t,2H,J＝6.8Hz),3.42(t,2H,J＝6.8Hz),2.44(s,3H),1.93-1.79(m,4H)。
Compound 203
4' -methyl-d 3-N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
A) N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) -4' - (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) biphenyl-3-carboxamide
To 4' -bromo-N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide (0.47g,0.98mmol), bis (pinacolato) diboron (0.37g,1.5mmol), potassium acetate (0.29g,2.9mmol), 1, 4-bis To a mixture of alkane (20mL) and dimethyl sulfoxide (0.2mL) was added [1,1' -bis (diphenylphosphino) ferrocene]Palladium (II) dichloride complex with dichloromethane (1:1) (24mg,0.029 mmol). The mixture was heated at 85 ℃ overnight under an argon atmosphere. After cooling, the mixture was filtered through Celite and the filter cake was washed with EtOAc. Concentrating the filtrate, passing through a silica gel column (0-10% EtOH/CH)2Cl2) The residue was purified to give a brown foam. LC-MS 526.2[ M +1 ]]+。
B)4' -methyl-d 3-N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide
K3PO4Solutions ofGive 2.0M K3PO4The aqueous solution was degassed and purged with argon.Borate solutionN- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) -4' - (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) biphenyl-3-carboxamide (100mg,0.19mmol) was dissolved in DMF (0.8 mL). The solution was degassed and purged with argon.Catalyst solutionReacting [1,1' -bis (diphenylphosphino) ferrocene]The complex of palladium (II) dichloride and dichloromethane (1:1) (10mg,0.012mmol) was dissolved well in degassed N, N-dimethylformamide (5.0 mL). The solution was degassed and purged with argon.Reaction ofAt room temperature and in an argon atmosphereTo a small reaction flask was added 1mL of orange-red catalyst solution (i.e., using 2mg of catalyst) and iodomethane-d 3(55mg,0.38 mmol). Then borate solution (0.19mmol) was added followed by 0.30mL of 2.0M K3PO4An aqueous solution. The resulting reaction mixture was heated in an oil bath at 100 ℃ for 20min under an argon atmosphere. LC-MS showed no remaining borate. After cooling, the reaction was quenched with 1.0mL MeOH, and the resulting solution was filtered and subjected to preparative HPLC (100X20.2mm, C18 column; 30-80% MeCN-water [10mM Et2NH](ii) a Flow rate 20 mL/min).
LC-MS:417.2[M+1]+；1H NMR(400MHz,DMSO-d6):9.25(t,1H,J＝6.0Hz),8.44(d,1H,J＝2.0Hz),8.21(t,1H,J＝1.6Hz),7.94(t,1H,J＝1.6Hz),7.89(t,1H,J＝1.6Hz),7.67(d,2H,J＝8.4Hz),7.63(dd,1H,J＝8.0,2.4Hz),7.31(d,2H,J＝8.4Hz),7.22(d,1H,J＝8.0Hz),4.48(d,2H,J＝6.0Hz),3.50(t,2H,J＝6.8Hz),3.42(t,2H,J＝6.4Hz),2.44(s,3H),1.93-1.78(m,4H)。
Compound 206
(S) -N- (2-hydroxy-1- (6-methoxypyridin-3-yl) ethyl) -4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxamide
To 4 '-methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (35mg,0.12mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (46mg,0.24mmol), 1-hydroxybenzotriazole hydrate (18mg,0.12mmol) and CH2Cl2(3mL) to the mixture was added (S) -2-amino-2- (6-methoxypyridin-3-yl) ethanol (30mg,0.18mmol) and N, N-diisopropylethylamine (42. mu.L, 0.24 mmol). The mixture was stirred at room temperature overnight and then concentrated. By preparative HPLC (100X20.2mm, C18 column; 30-80% CH3CN-Water [10mM Et2NH]) The residue was purified to give a white foam.
LC-MS:440.9[M+1]+；1H NMR(400MHz,CDCl3):8.32(m,1H),8.27-8.23(m,3H),7.65(dd,1H,J＝8.4,2.8Hz),7.53(d,2H,J＝8.4Hz),7.30(d,2H,J＝8.0Hz),7.16(d,1H,J＝7.2Hz),6.77(d,1H,J＝8.8Hz),5.26(m,1H),4.11-4.00(m,2H),3.93(s,3H),3.13(s,3H),2.42(s,3H)。
Compound 219
(R) -5- (cyclopentylsulfonyl) -4' -methyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) biphenyl-3-carboxamide
A) 3-bromo-5- (cyclopentylthio) benzoic acid
To a stirred mixture of sodium hydride (60% in mineral oil, 0.82g,20.5mmol) and DMSO (15mL) in a sealable flask was slowly added cyclopentanethiol (1.23mL,11.9mmol) at 0 ℃. The mixture was stirred at room temperature for 30min, then a solution of 3, 5-dibromobenzoic acid (2.5g,8.9mmol) in DMSO (10mL) was added. The reaction was sealed and stirred at 80 ℃ overnight. After cooling, the reaction mixture was poured into water, adjusted to pH5-6 with 15% HCl (aq) and extracted with EtOAc. The combined organic layers were washed with brine, anhydrous MgSO4Drying and concentration gave the crude product, which was used in the next reaction without further purification.
B) 3-bromo-5- (cyclopentylthio) benzoic acid methyl ester
A solution of 3-bromo-5- (cyclopentylthio) benzoic acid (1.49g,4.95mmol) in methanol (40mL) was treated with acetyl chloride (3.87mL,54.4mmol) at 0 deg.C. The reaction mixture was heated to reflux overnight and then concentrated under reduced pressure. The residue was purified by flash chromatography (0-20% EtOAc/hexanes) to give the title compound as a colorless oil.
C) 3-bromo-5- (cyclopentylsulfonyl) benzoic acid methyl ester
To 3-bromo-5- (cyclopentylthio) benzoic acid methyl ester at 0 deg.C(1.2g,3.8mmol) to a stirred solution in dichloromethane (20mL) was added m-chloroperbenzoic acid (70% purity, 2.35g,9.52mmol) slowly in portions. The mixture was slowly warmed to room temperature and stirred overnight. With Na2CO3The reaction mixture was washed with an aqueous solution and brine, and anhydrous MgSO4Drying and concentrating. The residue was purified by silica gel column (EtOAc/hexanes) to give a colorless oil.
D)5- (Cyclopentylsulfonyl) -4' -methylbiphenyl-3-carboxylic acid methyl ester
To a microwave vial was added a solution of methyl 3-bromo-5- (cyclopentylsulfonyl) benzoate (0.50g,1.44mmol), p-tolylboronic acid (0.44g,3.25mmol), toluene (7mL) and cesium carbonate (1.06g,3.26mmol) in water (0.3 mL). The vial was degassed, purged with argon, and then tetrakis (triphenylphosphine) palladium (0) (0.16g,0.14mmol) was added. The reaction mixture was subjected to microwave irradiation at 100 ℃ for 2 h. After cooling, the mixture was filtered and the filtrate was concentrated in vacuo. The residue was purified by flash column (0-25% EtOAc/hexanes) to give the title compound as a colorless oil.
E)5- (Cyclopentylsulfonyl) -4' -methylbiphenyl-3-carboxylic acid
To a solution of methyl 5- (cyclopentylsulfonyl) -4' -methylbiphenyl-3-carboxylate (0.34g,0.95mmol) in tetrahydrofuran (15mL) was added 2.5M aqueous lithium hydroxide (1.0mL,2.5 mmol). The reaction mixture was stirred at 60 ℃ overnight. The aqueous solution was acidified to pH 5 with 15% HCl (aq) and extracted with EtOAc. The combined organic layers were concentrated in vacuo to give the title compound as a white solid.
F) (R) -5- (cyclopentylsulfonyl) -4' -methyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) biphenyl-3-carboxamide
To a solution of 5- (cyclopentylsulfonyl) -4' -methylbiphenyl-3-carboxylic acid (50mg,0.14mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (40mg,0.29mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (140. mu.L, 0.80 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. The reaction mixture was purified by preparative HPLC to give the final product as a pale solid.
LC-MS:464.1[M+1]+；1H NMR(400MHz,DMSO-d6):9.26(d, 1H,J＝7.4Hz),8.75(s,2H),8.47(t,1H,J＝1.5Hz),8.29(t,1H,J＝1.5Hz),8.21(t,1H,J＝1.6Hz),7.74(d,2H,J＝8.2Hz),7.36(d,2H,J＝8.0Hz),5.22(m,1H),3.99(m,1H),2.60(s,3H),2.38(s,3H),2.00-1.75(m,4H),1.70-1.50(m,4H),1.57(d,3H,J＝7.1Hz)。
Compound 220
3- (5-methylpyridin-2-yl) -N- ((6-methylpyridin-3-yl) methyl) -5- (trifluoromethoxy) benzamide
A) 3-bromo-N- ((6-methylpyridin-3-yl) methyl) -5- (trifluoromethoxy) benzamide
To a 5mL microwave vial was added 1-bromo-3-iodo-5- (trifluoromethoxy) benzene (98mg,0.27mmol), (6-methylpyridin-3-yl) methylamine (50mg,0.41mmol), molybdenum hexacarbonyl (160mg,0.61mmol), palladium acetate (8mg,0.036mmol), 1, 8-diazabicyclo [5.4.0 mmol)]Undec-7-ene (120mg,0.79mmol) and 1, 4-bisAlkane (2 mL). The vial was sealed in a nitrogen atmosphere, and the reaction system was subjected to microwave irradiation at 120 ℃ for 20 minutes. After cooling, the mixture was purified by flash chromatography to give the desired product as a white solid. LC-MS 391.2[ M +1 ]]+；1H NMR(CDCl3,400MHz):8.43(d,J＝2.1Hz,1H),7.85(t,J＝1.5Hz,1H),7.63-7.61(m,2H),7.52(s,1H),7.16(d,J＝8.0Hz,1H),6.74(Br、1H),4.60(d,J＝5.8Hz,2H),2.55(s,3H)。
B)3- (5-methylpyridin-2-yl) -N- ((6-methylpyridin-3-yl) methyl) -5- (trifluoromethoxy) benzamide
A mixture of 3-bromo-N- ((6-methylpyridin-3-yl) methyl) -5- (trifluoromethoxy) benzamide (60mg,0.15mmol), 5-methyl-2- (tributylstannyl) pyridine (0.1mL,0.3mmol), tetrakis (triphenylphosphine) palladium (0) (20mg,0.02mmol) and toluene (4mL) was subjected to microwave irradiation at 120 ℃ for 2 hours under an argon atmosphere. LC-MS showed a strong peak of the starting material. The mixture was degassed with nitrogen and further subjected to microwave irradiation at 120 ℃ for 2 hours. The mixture was cooled to room temperature and concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM Et2NH]) The residue was purified to give the desired product as a white solid.
LC-MS:401.7[M+1]+；1H NMR(DMSO-d6,400MHz):9.36(t,J＝5.8Hz,1H),8.60(t,J＝1.4Hz,1H),8.56(d,J＝2.2Hz,1H),8.45(d,J＝1.9Hz,1H),8.21(s,1H),8.04(d,J＝8.1Hz,1H),7.85(s,1H),7.78(dd,J＝1.6,8.2Hz,1H),7.65(dd,J＝2.4,8.0Hz,1H),7.22(d,J＝8.0Hz,1H),4.49(d,J＝5.6Hz,2H),2.44(s,3H),2.36(s,3H)。
Compound 221
(S) -N- (2-hydroxy-1- (2-methylpyrimidin-5-yl) ethyl) -4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxamide
To 4 '-methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (35mg,0.12mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (46mg,0.24mmol), 1-hydroxybenzotriazole hydrate (18mg,0.12mmol) and CH2Cl2(3mL) to the mixture was added (S) -2- (tert-butyldimethylsilyloxy) -1- (2-methylpyrimidin-5-yl) ethylamine (48mg,0.18mmol) (WO2008/130481) and N, N-diisopropylethylamine (42. mu.L, 0.24 mmol). The mixture was stirred at room temperature overnight and then concentrated. The residue was dissolved in MeOH (5mL), concentrated, and HCl solution (0.5mL) was added. The mixture was stirred at room temperature for 2h, then concentrated in vacuo. By passingPreparative HPLC (100X20.2mm, C18 column; 30-80% MeCN-water [10mM Et2NH]) The residue was purified to give a white solid.
LC-MS:426[M+1]+；1H NMR(CDCl3,400MHz):8.78(s,2H),8.32(t,1H,J＝1.6Hz),8.30-8.25(m,2H),7.53(d,2H,J＝8.0Hz),7.47(d,1H,J＝7.2Hz),7.29(d,2H,J＝8.0Hz),5.30(m,1H),4.17(dd,1H,J＝10.8,4.0Hz),4.05(dd,1H,J＝10.8,4.8Hz),3.13(s,3H),2.73(s,3H),2.41(s,3H)。
Compound 224
2-methyl-5- ((4' -methyl-5- (pyrrolidine-1-carbonyl) biphenyl-3-ylcarboxamido) methyl) pyridine 1-oxide
To a 20mL reaction vial was added 4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (pyrrolidine-1-carbonyl) biphenyl-3-carboxamide (30mg,0.073mmol) and dichloromethane (1.0 mL). The mixture was cooled to 0 ℃ and an aqueous solution of m-chloroperbenzoic acid (70% purity, 28mg,0.11mmol) was added dropwise. The reaction mixture was stirred for another 3hr, then water was added. The aqueous layer was extracted with dichloromethane and the combined organic layers were dried and concentrated. The resulting solid was purified by preparative HPLC to give the title compound.
LC-MS:429.6[M+1]+。
Compound 225
(R) -3-methoxy-5- (5-methylpyridin-2-yl) -N- (1- (2-methylpyrimidin-5-yl) ethyl) benzamide
A) 3-bromo-5-methoxybenzoic acid
To a 3-neck flask were added 3, 5-dibromobenzoic acid (1.0g,3.6mmol), 4M sodium methoxide in methanol (1.2mL,4.8mmol), and N, N-dimethylformamide (1.2 mL). The mixture was heated at 110 ℃ and copper (I) bromide (51mg,0.36mmol) was added. The mixture was stirred at 110 ℃ over 4 days. After cooling, the mixture is poured into 1N aqueous HCl, adjusted to low pH and treated with CH2Cl2And (4) extracting. With Na2SO4The organic layer was dried and concentrated to dryness. The crude product was used in the next reaction without further purification. LC-MS 231.2[ M-1 ]]-。
B) 3-bromo-5-methoxybenzoic acid methyl ester
To crude 3-bromo-5-methoxybenzoic acid (40% purity, 0.8g,1.4mmol) in CH at 0 deg.C2Cl2To a stirred mixture (50mL) was added DMF (0.1mL) and acetyl chloride (0.2mL,3 mmol). The mixture was slowly warmed to rt and stirred overnight. The mixture was then cooled to 0 ℃ and methanol (2mL) was added slowly. The mixture was stirred at rt for 3h, then Na2CO3Washed with aqueous solution, brine and dried (Na)2SO4) And (4) concentrating. The residue was purified by flash chromatography to afford the desired product as a white solid.1H NMR(CDCl3,400MHz):7.76(t,J＝1.6Hz,1H),7.49(t,J＝1.3Hz,1H),7.24(t,J＝2.2Hz,1H),3.92(s,3H),3.84(s,3H)。
C) 3-methoxy-5- (5-methylpyridin-2-yl) benzoic acid methyl ester
A mixture of methyl 3-bromo-5-methoxybenzoate (0.1g,0.4mmol), 5-methyl-2- (tributylstannyl) pyridine (0.1mL,0.3mmol), tetrakis (triphenylphosphine) palladium (0) (20mg,0.02mmol) and toluene (4mL) was subjected to microwave irradiation at 120 ℃ for 2 hours under an argon atmosphere. The mixture was cooled to room temperature and purified by flash chromatography to afford the desired product as a light yellow oil. LC-MS 257.9[ M +1 ]]+。
D) 3-methoxy-5- (5-methylpyridin-2-yl) benzoic acid
3-methoxy-5- (5-Methyl methylpyridin-2-yl) benzoate (80mg,0.2mmol), barium hydroxide octahydrate (300mg,0.95mmol) and methanol (10 mL). The mixture was stirred at room temperature overnight. 2N HCl (ether solution, 30mL) was added and the volatiles were removed under reduced pressure. LC-MS 244.1[ M +1 ]]+。
E) (R) -3-methoxy-5- (5-methylpyridin-2-yl) -N- (1- (2-methylpyrimidin-5-yl) ethyl) benzamide
To a round bottom flask was added 3-methoxy-5- (5-methylpyridin-2-yl) benzoic acid (26mg,0.11mmol), (R) -1- (2-methylpyrimidin-5-yl) ethylamine (29mg,0.21mmol), N- (3-dimethylaminopropyl) -N' -ethylcarbodiimide hydrochloride (41mg,0.21mmol), 1-hydroxybenzotriazole hydrate (33mg,0.21mmol), triethylamine (22mg,0.21mmol) and dichloromethane (3 mL). The mixture was stirred at room temperature overnight and then concentrated. By preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM Et2NH]) The residue was purified to give the desired product as a white solid.
LC-MS:363.1[M+1]+；1H NMR(400MHz,CD3OD):8.65(s,1H),8.60(s,1H),8.38(d,J＝1.9Hz,1H),7.87(t,J＝1.5Hz,1H),7.72(d,J＝8.1Hz,1H),7.65(dd,J＝1.6,8.1Hz,1H),7.58(dd,J＝1.6,2.4Hz,1H),7.34(dd,J＝1.6,2.4Hz,1H),5.17(q,J＝7.1Hz,1H),3.82(s,3H),2.57(s,3H),2.31(s,3H),1.55(d,J＝7.1Hz,3H)。
Compound 227
5- (hydroxy (phenyl) methyl) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
A)5- (hydroxymethyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
To 5- (ethoxycarbonyl) -4' -methylbiphenyl-3-carboxylic acid (2.66g,8.89mmol) in dry tetrahydrofuran (40mL) at 0 ℃ under a nitrogen atmosphereTo the stirred solution in (1M) borane in tetrahydrofuran (20mL,20mmol) was added. After the addition was complete, the reaction was stirred at 0 ℃ for 30 minutes and then the cooling bath was removed. After stirring at room temperature for 2 hours, the reaction mixture was cooled at 0 ℃ and quenched by addition of 2M aqueous HCl (30 mL). The mixture was stirred at room temperature for 1 hour, then extracted with EtOAc (50mL × 3). The combined organic layers were washed with brine, anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by silica gel column to give the title compound.
B) 5-formyl-4' -methylbiphenyl-3-carboxylic acid ethyl ester
A mixture of ethyl 5- (hydroxymethyl) -4' -methylbiphenyl-3-carboxylate (400mg,1.5mmol) and manganese (IV) oxide (660mg,6.6mmol) in dichloromethane (14mL) was stirred at room temperature overnight. The reaction mixture was filtered through Celite, washing with dichloromethane. The filtrate was concentrated to give the title compound.
C)5- (hydroxy (phenyl) methyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
Phenylmagnesium bromide (850mg,4.7mmol) was added to a solution of ethyl 5-formyl-4' -methylbiphenyl-3-carboxylate (360mg,1.3mmol) in tetrahydrofuran (8mL) at 0 ℃. The reaction mixture was stirred at room temperature for 3h, then water (10mL) was added. The mixture was extracted with EtOAc (20mLx 2). With anhydrous MgSO4The combined organic layers were dried and concentrated. The residue was purified by silica gel column chromatography to give the title compound.
D)5- (hydroxy (phenyl) methyl) -4' -methylbiphenyl-3-carboxylic acid
To a solution of ethyl 5- (hydroxy (phenyl) methyl) -4' -methylbiphenyl-3-carboxylate (230mg,0.66mmol) in 1, 4-bisTo a solution in alkane (2mL) and water (1mL) was added lithium hydroxide (48mg,2 mmol). The reaction mixture was stirred at 60 ℃ overnight. After cooling, the mixture was acidified to pH 5 with 15% HCl (aq) and extracted with EtOAc. Drying the organic layer and concentrating under vacuumThe title compound was obtained as a white solid.1H NMR (300MHz, acetone-d 6):8.14(t,1H, J ═ 2.0Hz),8.09(m,1H),7.97(m,1H),7.60-7.47(m,4H),7.37-7.19(m,5H),6.00(s,1H),2.37(s, 3H).
E)5- (hydroxy (phenyl) methyl) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a solution of 5- (hydroxy (phenyl) methyl) -4' -methylbiphenyl-3-carboxylic acid (50mg,0.16mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (70mg,0.34mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (500. mu.L, 2.87 mmol). The reaction mixture was stirred at 30 ℃ for 16 hours. By preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM Et2NH]) The reaction mixture was purified to give a pale solid.
LC-MS:437.2[M+1]+；1H NMR(400MHz,DMSO-d6):8.92(d,1H,J＝7.8Hz),8.47(d,1H,J＝2.1Hz),7.98(bs,1H),7.83(bs,1H),7.79(bs,1H),7.67(dd,1H,J＝8.0,2.2Hz),7.59(d,2H,J＝8.1Hz),7.42(d,2H,J＝7.3Hz0,7.35-7.25(m,4H),7.23-7.15(m,2H),6.04(m,1H),5.82(d,1H,J＝4.0Hz),5.17(m,1H),2.43(s,3H),2.35(s,3H),1.50(d,3H,J＝7.1Hz)。
Compound 229
(R) -5-benzoyl-4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a solution of 5- (hydroxy (phenyl) methyl) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide (20mg,0.04mmol) in dichloromethane (2.0mL) was added Dess-Martin periodinane (26mg,0.061 mmol). The reaction mixture was stirred at 40 ℃ for 2 hours and LC-MS showed the reaction was complete. By CH2Cl2The reaction solution was diluted with 10% Na2S2O3Saturated NaHSO3Aqueous solution and brine. With anhydrous MgSO4The organic layer was dried and concentrated. The residue was purified by silica gel column to give a white solid.
LC-MS:435.3[M+1]+；1H NMR(400MHz,DMSO-d6):9.13(d,1H,J＝7.7Hz),8.48(d,1H,J＝2.2Hz),8.42(t,1H,J＝1.5Hz),8.16(bs,1H),8.06(t,1H,J＝1.5Hz),7.85-7.55(m,8H),7.33(d,2H,J＝8.1Hz),7.21(d,1H,J＝8.0Hz),5.20(m,1H),2.43(s,3H),2.37(s,3H),1.52(d,3H,J＝7.1Hz)。
Compound 232
(R) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) -5- (pyrimidin-2-yloxy) biphenyl-3-carboxamide
A) 3-bromo-5-hydroxy-benzoic acid methyl ester
To a solution of 3-bromo-5-hydroxybenzoic acid (3.41g,15mmol) (J.chem.Soc.1955,463) in dry methanol (80mL) was added acetyl chloride (2.56mL,36mmol) at 0 ℃. The mixture was then heated at reflux overnight. The volatiles were removed in vacuo to afford the title compound.
B) 5-hydroxy-4' -methylbiphenyl-3-carboxylic acid methyl ester
To a stirred solution of methyl 3-bromo-5-hydroxy-benzoate (4.09g,17.7mmol) and p-tolylboronic acid (2.86g,21mmol) in toluene (36mL) at room temperature was added a solution of cesium carbonate (7.14g,22mmol) in water (3.6 mL). The mixture was purged with nitrogen and tetrakis (triphenylphosphine) palladium (0) (1.05g,0.906mmol) was added. The reaction mixture was subjected to microwave irradiation at 110 ℃ for 1 hour. After cooling, the mixture was filtered through Celite and the filtrate was concentrated. The residue was purified by flash chromatography to give the title compound.
C)4' -methyl-5- (pyrimidin-2-yloxy) biphenyl-3-carboxylic acid methyl ester
To a stirred solution of methyl 5-hydroxy-4' -methylbiphenyl-3-carboxylate (300mg,1.24mmol) in tetrahydrofuran (20mL) was added sodium hydride (60% solution in mineral oil, 55mg,1.37mmol) at 0 ℃ over a 10 minute period. The reaction mixture was concentrated to give a green solid. To a microwave vial was added the resulting green solid, dimethyl sulfoxide (8mL) and 2-chloropyrimidine (118mg,0.98 mmol). The mixture was subjected to microwave irradiation at 100 ℃ for 4 minutes. After cooling, the mixture was filtered through Celite and the filtrate was concentrated. The residue was purified by flash chromatography to give the title compound.
D)4' -methyl-5- (pyrimidin-2-yloxy) biphenyl-3-carboxylic acid
To a stirred solution of methyl 4' -methyl-5- (pyrimidin-2-yloxy) biphenyl-3-carboxylate (60mg,0.19mmol) in tetrahydrofuran (3mL) was added 2.5M aqueous lithium hydroxide (0.77mL,1.9 mmol). The mixture was stirred at rt overnight, then acidified to pH 5 by addition of 2N aqueous HCl and extracted with EtOAc (50 mL). The organic layer was separated, dried over sodium sulfate, filtered, and concentrated to give the title product.1H NMR(CD3OD,400MHz):8.63(d,2H,J＝4.8Hz),8.16(t,1H,J＝1.6Hz),7.74(t,1H,J＝2.0Hz),7.65(t,1H,J＝2.0Hz),7.56(d,2H,J＝8.0Hz),7.29(d,2H,J＝8.0Hz),7.24(t,1H,J＝4.8Hz),2.39(s,3H)。
E) (R) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) -5- (pyrimidin-2-yloxy) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (pyrimidin-2-yloxy) biphenyl-3-carboxylic acid (20mg,0.065mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (30mg,0.143mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (60mg,0.16mmol), and N, N-diisopropylethylamine (200 μ L,1.15 mmol). The reaction mixture was stirred at room temperature for 16 hours. LC-MS showed the reaction was complete. By preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM ET2NH]) Purification of the reactionThe mixture was obtained as a pale solid.
LC-MS:425.1[M+1]+；1H NMR(DMSO-d6,400MHz):8.97(d, 1H,J＝7.8Hz),8.67(d,2H,J＝4.8Hz),8.47(d,1H,J＝2.2Hz),8.06(t,1H,J＝1.4Hz),7.70-7.62(m,5H),7.32-7.25(m,3H),7.21(d,1H,J＝8.0Hz),5.18(m,1H),2.43(s,3H),2.36(s,3H),1.51(d,3H,J＝7.1Hz)。
Compound 236
(R) -4' -methyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (thiazol-2-yloxy) biphenyl-3-carboxamide
A)4' -methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylic acid methyl ester
A mixture of methyl 5-hydroxy-4' -methylbiphenyl-3-carboxylate (100mg,0.41mmol), potassium carbonate (114mg,0.83mmol), 2-bromothiazole (0.38mL,4.2mmol) and dimethyl sulfoxide (2.5mL) was stirred at 135 deg.C overnight. After cooling, the mixture was diluted with EtOAc and washed with NaHCO3Washed with aqueous (saturated) solution over anhydrous MgSO4Drying and concentrating. The residue was purified by flash chromatography (0-20% EtOAc/hexanes) to give the title compound.
B)4' -methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylic acid
To a stirred solution of methyl 4' -methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylate (560mg,1.6mmol) in tetrahydrofuran (20mL) was added 2.5M aqueous lithium hydroxide (6.7mL,17 mmol). After stirring at rt overnight, the mixture was acidified to pH 5 by addition of 2N aqueous HCl and extracted with EtOAc. The organic layer was dried over sodium sulfate and concentrated to give the title product.1H NMR(CD3OD,400MHz):8.18(t,1H,J＝1.6Hz),7.82(dd,1H,J＝2.4,1.2Hz),7.74(t,1H,J＝2.0Hz),7.57(d,2H,J＝8.0Hz),7.30(d,2H,J＝8.4Hz),7.28(d,1H,J＝4.0Hz),7.13(d,1H,J＝4.0Hz),2.39(s,3H)。
C) (R) -4' -methyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (thiazol-2-yloxy) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylic acid (50mg,0.16mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (45mg,0.33mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (150mg,0.39mmol), and N, N-diisopropylethylamine (200 μ L,1.15 mmol). The reaction mixture was stirred at room temperature for 16 hours. LC-MS showed the reaction was complete. By preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM Et2NH]) The reaction mixture was purified to give a pale solid.
LC-MS:431.2[M+1]+；1H NMR(400MHz,DMSO-d6):9.07(d,1H,J＝7.4Hz),8.73(s,2H),8.10(t,1H,J＝1.4Hz),7.82(t,1H,J＝2.0Hz),7.76(t,1H,J＝1.5Hz),7.68(d,2H,J＝8.2Hz),7.35-7.25(m,4H),5.18(m,1H),2.59(s,3H),2.36(s,3H),1.55(d,3H,J＝7.1Hz)。
Compound 240
5- (hydroxy (pyridin-2-yl) methyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
A)5- (hydroxy (pyridin-2-yl) methyl) -4' -methylbiphenyl-3-carboxylic acid ethyl ester
Grignard reagent was generated by treating magnesium (100mg,4.3mmol) with a solution of 2-bromopyridine (600mg,3.8mmol) and iodine (30mg) in tetrahydrofuran (5mL) at 60 ℃ for 1 hour under a nitrogen atmosphere. The mixture was cooled at 0 ℃ and ethyl 5-formyl-4' -methylbiphenyl-3-carboxylate (332mg,1.24mmol) was added. The reaction mixture was stirred at 0 ℃ for 2H, then by addition of 3mL of 2N H2SO4The aqueous solution was quenched. The mixture was extracted with EtOAc (20mL x2)A compound (I) is provided. The combined organic layers were washed with 1N aqueous NaOH, brine, and anhydrous MgSO4Drying, filtering and concentrating. The residue was chromatographed to give a yellow oil.
B)5- (hydroxy-pyridin-2-yl-methyl) -4' -methyl-biphenyl-3-carboxylic acid
Ethyl 5- (hydroxy (pyridin-2-yl) methyl) -4' -methylbiphenyl-3-carboxylate (319mg,0.92mmol) was dissolved in 1, 4-bisAlkane (6mL) and water (0.3 mL). Lithium hydroxide (120mg,5.0mmol) was added to the stirred solution and the reaction mixture was stirred at room temperature until the reaction was complete. Cooling the mixture with 2N H2SO4The aqueous solution was acidified to pH5-6 and extracted with EtOAc (20mL x 3). The combined organic layers were washed with brine and dried (MgSO)4) And concentrated to give a pale yellow solid.1H NMR(400MHz,CD3OD):8.47(m,1H),8.13(t,1H,J＝1.6Hz), 8.02(t,1H,J＝1.6Hz),7.91(t,1H,J＝1.6Hz),7.87(td,1H,J＝7.6,1.6Hz),7.71(d,1H,J＝8.0Hz),7.51(d,2H,J＝8.0Hz),7.31(ddd,1H,J＝7.6,5.2,1.2Hz),7.27(d,2H,J＝8.0Hz),5.93(s,1H),2.37(s,3H)。
C)5- (hydroxy (pyridin-2-yl) methyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
To a solution of 5- (hydroxy-pyridin-2-yl-methyl) -4 '-methyl-biphenyl-3-carboxylic acid (35mg,0.11mmol) in N, N-dimethylformamide (1mL) were added (6-methylpyridin-3-yl) methylamine (40mg,0.33mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol) and N, N-diisopropylethylamine (100 μ L,0.57 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. By preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM Et2NH]) The reaction mixture was purified to give a pale solid.
LC-MS:424.1[M+1]+；1H NMR(400MHz,DMSO-d6):9.15(t,1H,J＝5.5Hz),8.46(d,1H,J＝4.0Hz),8.41(bs,1H),7.99(bs,1H),7.88(bs,1H),7.83(bs,1H),7.79(t,1H,J＝7.8Hz),7.65-7.55(m,4H),7.30(d,2H,J＝7.6Hz),7.22(m,2H),6.26(bs,1H),5.83(bs,1H),4.45(d,2H,J＝5.4Hz),2.43(s,3H),2.35(s,3H)。
Compound 241
5- (hydroxy (pyridin-2-yl) methyl) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a solution of 5- (hydroxy-pyridin-2-yl-methyl) -4 '-methyl-biphenyl-3-carboxylic acid (25mg,0.078mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (35mg,0.17mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (50mg,0.13mmol), and N, N-diisopropylethylamine (200 μ L,1.15 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then passed through preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM Et2NH]) Purification gave a pale solid.
LC-MS:438.1[M+1]+；1H NMR(400MHz,DMSO-d6):8.92(d,1H,J＝7.8Hz),8.45(bs,2H),7.98(bs,1H),7.85(bs,1H),7.81(bs,1H),7.79(t,1H,J＝7.2Hz),7.70-7.55(m,4H),7.30(d,2H,J＝8.0Hz),7.22(m,2H),6.25(bs,1H),5.83(s,1H),5.20-5.10(m,1H),2.43(s,3H),2.35(s,3H),1.50(d,3H,J＝7.1Hz)。
Compound 242
(R) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) -5- (pyridin-2-yloxy) biphenyl-3-carboxamide
A)4' -methyl-5- (pyridin-2-yloxy) biphenyl-3-carboxylic acid methyl ester
A mixture of methyl 5-hydroxy-4' -methylbiphenyl-3-carboxylate (100mg,0.41mmol), potassium carbonate (100mg,0.72mmol), 2-bromopyridine (0.40mL,4.2mmol), and dimethyl sulfoxide (2mL) was stirred at 135 deg.C overnight. After cooling, the mixture was diluted with EtOAc and washed with NaHCO3Washed with aqueous (saturated) solution over anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by flash chromatography (0-20% EtOAc/hexanes) to give the title compound.
B)4' -methyl-5- (pyridin-2-yloxy) biphenyl-3-carboxylic acid
To a stirred solution of methyl 4' -methyl-5- (pyridin-2-yloxy) biphenyl-3-carboxylate (460mg,1.4mmol) in tetrahydrofuran (20mL) was added 2.5M aqueous lithium hydroxide (5.6mL,14 mmol). After stirring at rt overnight, the mixture was acidified to pH 5 by addition of 2N aqueous HCl and extracted with EtOAc. The organic layer was dried over sodium sulfate, filtered, and concentrated to give the title compound.1H NMR(CD3OD,300MHz):8.17(d,1H,J＝3.2Hz),8.11(s,1H),7.88(td,1H,J＝8.0,2.0Hz),7.65(s,1H),7.58-7.52(m,3H),7.29(d,2H,J＝8.4Hz),7.16(dd,1H,J＝7.2,4.8Hz),7.06(d,1H,J＝8.0Hz),2.38(s,3H)。
C) (R) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) -5- (pyridin-2-yloxy) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (pyridin-2-yloxy) biphenyl-3-carboxylic acid (50mg,0.16mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (70mg,0.34mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (150mg,0.39mmol), and N, N-diisopropylethylamine (400 μ L,2.30 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the final product as a pale solid.
LC-MS:424.0[M+1]+；1H NMR(400MHz,DMSO-d6):8.97(d,1H,J＝7.7Hz),8.47(bs,1H),8.15(d,1H,J＝4.2Hz),8.02(bs,1H),7.89(t,1H,J＝8.5Hz),7.70-7.60(m,3H),7.57(bs,2H),7.30(d,2H,J＝8.0Hz),7.25-7.05(m,3H),5.18(m,1H),2.43(s,3H),2.35(s,3H),1.51(d,3H,J＝7.1Hz)。
Compound 248
5- (2-methoxyethoxy) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
A)5- (2-methoxyethoxy) -4' -methylbiphenyl-3-carboxylic acid methyl ester
A mixture of methyl 5-hydroxy-4' -methylbiphenyl-3-carboxylate (400mg,1.65mmol), 1-bromo-2-methoxyethane (436mg,3.14mmol), potassium carbonate (434mg,3.14mmol) and dimethyl sulfoxide (5mL) was stirred at 135 deg.C overnight. After cooling, the mixture was diluted with EtOAc and washed with NaHCO3Washed with aqueous (saturated) solution over anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by flash chromatography (0-100% EtOAc/hexanes) to give the title compound.
B)5- (2-methoxyethoxy) -4' -methylbiphenyl-3-carboxylic acid
To a stirred solution of methyl 5- (2-methoxyethoxy) -4' -methylbiphenyl-3-carboxylate (703mg,2.34mmol) in tetrahydrofuran (40mL) was added 2.5M aqueous lithium hydroxide (9.6mL,24 mmol). After stirring at rt overnight, the mixture was acidified to pH 5 by addition of 2N HCl and extracted with EtOAc. Drying the organic layer (Na)2SO4) And concentrated to give the title compound.1H NMR(CD3OD,400MHz):7.84(t,1H,J＝1.6Hz), 7.55-7.51(m,3H),7.34(t,1H,J＝2.0Hz),7.27(d,2H,J＝8.0Hz),4.22(m,2H),3.79(m,2H),3.45(s,3H),2.38(s,3H)。
C)5- (2-methoxyethoxy) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
To a solution of 5- (2-methoxyethoxy) -4' -methylbiphenyl-3-carboxylic acid (50mg,0.18mmol) in N, N-dimethylformamide (1mL) were added (6-methylpyridin-3-yl) methylamine (45mg,0.37mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (150mg,0.39mmol), and N, N-diisopropylethylamine (150. mu.L, 0.86 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. LC-MS showed the reaction was complete. The reaction mixture was purified by preparative HPLC to give the title product.
LC-MS:391.5[M+1]+；1H NMR(400MHz,DMSO-d6):9.13(t,1H,J＝5.8Hz),8.42(d,1H,J＝2.0Hz),7.74(bs,1H),7.66-7.60(m,3H),7.40(bs,1H),7.34(bs,1H),7.29(d,2H,J＝8.0Hz),7.22(d,1H,J＝8.0Hz),4.47(d,2H,J＝5.8Hz),4.22(t,2H,J＝4.4Hz),3.69(t,2H,J＝4.4Hz),3.32(s,3H),2.44(s,3H),2.35(s,3H)。
Compound 254
(R) -2 '-cyano-4' -methyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (thiazol-2-yloxy) biphenyl-3-carboxamide
A)2 '-cyano-5-hydroxy-4' -methylbiphenyl-3-carboxylic acid methyl ester
To the flask, 5-methyl-2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzonitrile (2.1g,8.2mmol), methyl 3-bromo-5-hydroxy-benzoate (1.5g,6.2mmol), potassium phosphate (2.8g,13mmol), tetrakis (triphenylphosphine) palladium (0) (400mg,0.35mmol), 1, 4-bis (triphenylphosphine) were added under a nitrogen atmosphere Alkane (90mL) and water (20 mL). The reaction mixture was stirred at 85 ℃ overnight. After cooling, the mixture was filtered through Celite and the filter cake was washed with EtOAc. The filtrate was concentrated and the residue was purified by silica gel column to give the title compound.
B)2 '-cyano-4' -methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylic acid methyl ester
A stirred mixture of methyl 2 '-cyano-5-hydroxy-4' -methylbiphenyl-3-carboxylate (350mg,1.2mmol), potassium carbonate (363mg,2.63mmol), 2-bromothiazole (431mg,2.63mmol) and dimethyl sulfoxide (10mL) was heated at 135 deg.C overnight. After cooling, the reaction mixture was diluted with EtOAc. With NaHCO3The organic phase was washed with aqueous (saturated) solution and anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by flash chromatography to give the title compound.
C)2 '-cyano-4' -methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylic acid
To a solution of methyl 2 '-cyano-4' -methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylate (112mg,0.32mmol) in tetrahydrofuran (2mL) was added 2.5M aqueous lithium hydroxide solution (0.72mL,1.8 mmol). The reaction mixture was stirred at 60 ℃ overnight. The aqueous solution was acidified to pH 5 with 15% HCl (aq) and extracted with EtOAc. The combined organic layers were concentrated in vacuo to give the title compound as a white solid.
D) (R) -2 '-cyano-4' -methyl-N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (thiazol-2-yloxy) biphenyl-3-carboxamide
To a solution of 2 '-cyano-4' -methyl-5- (thiazol-2-yloxy) biphenyl-3-carboxylic acid (30mg,0.089mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (30mg,0.22mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol), and N, N-diisopropylethylamine (100mg,0.26mmol)μL,0.57 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours and then purified by preparative HPLC to give a brown solid.
LC-MS:455.9[M+1]+；1H NMR (400MHz, DMSO-d6):9.07(d,1H, J ═ 7.4Hz),8.71(s,2H),8.00(bs,1H),7.94(t,1H, J ═ 1.8Hz),7.83(bs,1H),7.76(t,1H, J ═ 1.8Hz),7.68-7.60(m,2H),7.32 and 7.30(AB,2H, J ═ 3.7Hz),5.17(m,1H),2.59(s,3H),2.41(s,3H),1.53(d,3H, J ═ 7.1 Hz).
Compound 256
2-methyl-5- ((4' -methyl-5- (methylsulfonyl) biphenyl-3-ylcarboxamido) methyl) pyridine 1-oxide
A20 mL reaction vial was charged with 4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (methylsulfonyl) biphenyl-3-carboxamide (40mg,0.10mmol), m-chloroperbenzoic acid (70% purity, 52mg,0.21mmol), dichloromethane (3mL), and water (1 mL). The reaction mixture was stirred at room temperature for 12 h. Saturated aqueous sodium bicarbonate was added and the layers were separated. The organic layer was dried over sodium sulfate and concentrated in vacuo. The resulting oil was purified by flash chromatography (0-10% methanol/dichloromethane) to give the title compound as a white solid.
LC-MS:411.2[M+1]+；1H NMR(400MHz,DMSO-d6):9.5(t,1H,J＝5.8Hz),8.47(t,1H,J＝1.5Hz),8.34(t,1H,J＝1.5Hz),8.29(t,1H,J＝1.5Hz),8.27(s,1H),7.75(d,2H,J＝8.1Hz),7.49(d,1H,J＝8.1Hz),7.36(d,2H,J＝8.1Hz),7.26(d,1H,J＝8.5Hz),4.89(d,2H,J＝5.8Hz),3.37(s,3H),2.82(s,3H),2.32(s,3H)。
Compound 257 (Novak)
(S) -N- (2-hydroxy-1- (6-methylpyridin-3-yl) ethyl) -4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxamide
To a solution of 4' -methyl-5- (methylsulfonyl) biphenyl-3-carboxylic acid (37mg,0.12mmol) in N, N-dimethylformamide (1mL) were added (S) -2- (tert-butyldimethylsilyloxy) -1- (6-methylpyridin-3-yl) ethylamine (80mg,0.27mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylEthylamine (120. mu.L, 0.69 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours. LC-MS showed the reaction was complete. To the reaction mixture was added MeOH (2mL) and 12N aqueous HCl (0.5 mL). The mixture was stirred at rt for 2hr, then concentrated in vacuo. By preparative HPLC (100X21.2mm C18 column, MeCN-water [10mM Et2NH]) The residue was purified to give the final product as a pale solid.
LC-MS:425[M+1]+；1H NMR(400MHz,DMSO-d6):9.16(d,1H, J＝8.0Hz),8.49(d,1H,J＝2.1Hz),8.47(t,1H,J＝1.6Hz),8.35(t,1H,J＝1.6Hz),8.28(t,1H,J＝1.6Hz),7.75(d,2H,J＝8.2Hz),7.71(dd,1H,J＝8.0,2.2Hz),7.37(d,2H,J＝8.0Hz),7.22(d,1H J＝8.0Hz),5.20-5.05(m,2H),3.85-3.65(m,2H),3.35(s,3H),2.44(s,3H),2.39(s,3H)。
Compound 264
(R) -3- (5-methylpyridin-2-yl) -N- (1- (6-methylpyridin-3-yl) ethyl) -5- (thiazol-2-yloxy) benzamide
A) 3-hydroxy-5- (5-methylpyridin-2-yl) benzoic acid methyl ester
A mixture of 3-bromo-5-hydroxy-benzoic acid methyl ester (1.47g,6.36mmol), 5-methyl-2- (tributylstannyl) pyridine (3.0g,7.85mmol), tetrakis (triphenylphosphine) -palladium (0) (300mg,0.26mmol) in toluene (20mL) and DMF (2mL) was subjected to microwave irradiation at 110 ℃ for 1 hour. After cooling, the mixture was diluted with water and extracted with EtOAc. With anhydrous MgSO4The combined organic layers were dried and concentrated in vacuo. The residue was purified by flash chromatography to give the title compound as a white solid.
B)3- (5-methylpyridin-2-yl) -5- (thiazol-2-yloxy) benzoic acid methyl ester
Reacting methyl 3-hydroxy-5- (5-methylpyridin-2-yl) benzoate (290mg,1.2mmol), 2-bromothiazole (410mg,2.5mmol),A stirred mixture of potassium carbonate (346mg,2.5mmol) and dimethyl sulfoxide (5mL) was heated at 135 deg.C overnight. The reaction mixture was then cooled to room temperature and diluted with EtOAc. With NaHCO3The organic phase was washed with aqueous (saturated) solution and anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by flash chromatography (0-30% EtOAc/hexanes) to give the title compound.
C)3- (5-methylpyridin-2-yl) -5- (thiazol-2-yloxy) benzoic acid
To a solution of methyl 3- (5-methylpyridin-2-yl) -5- (thiazol-2-yloxy) benzoate (240mg, 0.74mmol) in tetrahydrofuran (6mL) was added 2.5M aqueous lithium hydroxide (1.6mL,4.1 mmol). The reaction mixture was stirred at 60 ℃ overnight. The aqueous solution was acidified to pH 5-6 with 15% HCl (aq) and extracted with EtOAc. The combined organic layers were concentrated in vacuo to give the title compound as a white solid.1H NMR(CDCl3,400MHz):8.77(s,1H),8.70(s,1H),8.08(s,1H),8.04(s,1H),7.71(d,J＝8.0Hz,1H),7.64(dd,J＝8.0,2.0Hz,1H),7.25(d,J＝4.8Hz,1H),6.85(d,J＝4.0Hz,1H),2.41(s,3H)。
D) (R) -3- (5-methylpyridin-2-yl) -N- (1- (6-methylpyridin-3-yl) ethyl) -5- (thiazol-2-yloxy) benzamide
To a solution of 3- (5-methylpyridin-2-yl) -5- (thiazol-2-yloxy) benzoic acid (40mg,0.13mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (400. mu.L, 2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the final product as a light brown solid.
LC-MS:431[M+1]+；1H NMR (400MHz, DMSO-d6):9.08(d,1H, J ═ 7.8Hz),8.55(d,1H, J ═ 1.9Hz),8.49(m,2H),8.19(t,1H, J ═ 2.0Hz),8.01(d,1H, J ═ 8.2Hz),7.85(t,1H, J ═ 1.5Hz),7.77(dd,1H, J ═ 8.2,1.8Hz),7.69(dd,1H, J ═ 8.0,2.4Hz),7.33, and 7.30(AB,2H, J ═ 3.8Hz),7.21(d,1H, J ═ 7.0 ═ 8Hz),7.21(d,1H, J ═ 8.0 Hz), and 7.30 (J ═ b,2H, J ═ 3.8Hz)8.0Hz),5.19(m,1H),2.43(s,3H),2.36(s,3H),1.52(d,3H,J＝7.1Hz)。
Compound 269
(R) -3- (5-methylpyridin-2-yl) -N- (1- (2-methylpyrimidin-5-yl) ethyl) -5- (thiazol-2-yloxy) benzamide
To a solution of 3- (5-methylpyridin-2-yl) -5- (thiazol-2-yloxy) benzoic acid (40mg,0.13mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (40mg,0.29mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (120. mu.L, 0.69 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the product as a pale solid.
LC-MS:432.2[M+1]+；1H NMR (400MHz, DMSO-d6):9.12(d,1H, J ═ 7.5Hz),8.73(s,2H),8.55(d,1H, J ═ 2.0Hz),8.49(t,1H, J ═ 1.4Hz),8.19(t,1H, J ═ 2.1Hz),8.01(d,1H, J ═ 8.2Hz),7.84(bs,1H),7.77(dd,1H, J ═ 8.1,1.7Hz),7.33 and 7.30(AB,2H, J ═ 3.76Hz),5.19(m,1H),2.59(s,3H),2.36(s,3H),1.56(d,3H, J ═ 7.1 Hz).
Compound 274
(R) -3- (methoxymethyl) -5- (5-methylpyridin-2-yl) -N- (1- (2-methylpyrimidin-5-yl) ethyl) benzamide
A)5- (5-methylpyridin-2-yl) isophthalate dimethyl ester
Under a nitrogen atmosphere, dimethyl 5-bromoisophthalate (1.0g,3.66mmol) and 5-methyl were addedA mixture of 2- (tributylstannyl) pyridine (1.5mL,4.5mmol), tetrakis (triphenylphosphine) palladium (0) (200mg,0.17mmol), and toluene (10mL) was subjected to microwave irradiation at 120 ℃ for 2 hours. The mixture was cooled to room temperature and the precipitate was collected by filtration and washed with hexane to give a white solid as the desired product. The filtrate was purified by flash chromatography to give a further batch of the desired product (0.8 g total). LC-MS 286.3[ M +1 ]]+；1H NMR(400MHz,CDCl3):8.84(d,J＝1.6Hz,2H),8.71(t,J＝1.6Hz,1H),8.56(t,J＝0.8Hz,1H),7.75(d,J＝8.0Hz,1H),7.61(dd,J＝2.0,8.1Hz,1H),3.98(s,6H),2.41(s,3H)。
B)3- (hydroxymethyl) -5- (5-methylpyridin-2-yl) benzoic acid methyl ester
To a stirred solution of dimethyl 5- (5-methylpyridin-2-yl) isophthalate (1.78g,5.93 mmol) in methanol (100mL) was added sodium tetrahydroborate (2.0g,0.053mol) at 0 ℃. After 3h, saturated NH was used4The reaction was treated with aqueous Cl and the volatiles were removed in vacuo. The residue was dissolved in water by addition of NaHCO3Basified to pH 9 and then treated with CH2Cl2(70mL x 3). With anhydrous MgSO4The combined organic layers were dried and concentrated. The residue was purified by flash chromatography to give the title compound.
C)3- (methoxymethyl) -5- (5-methylpyridin-2-yl) benzoic acid methyl ester
To a stirred solution of methyl 3- (hydroxymethyl) -5- (5-methylpyridin-2-yl) benzoate (110mg,0.41mmol) in N, N-dimethylformamide (2mL) was added sodium hydride (60% solution in mineral oil, 21mg,0.52mmol) at 0 deg.C. After 30 min, methyl iodide (73mg,0.51mmol) was added at 0 ℃. The mixture was warmed to room temperature and stirred for 30 minutes. The mixture was quenched with water (2mL) and extracted with EtOAc. With anhydrous MgSO4The combined organic layers were dried and concentrated to give the title compound.
D)3- (methoxymethyl) -5- (5-methylpyridin-2-yl) benzoic acid
To 3- (methoxymethyl) -5- (5-methylpyridine-2-yl) benzoic acid methyl ester (75mg,0.28mmol) in 1, 4-bisTo a stirred solution of alkane (3mL) and water (0.3mL) was added lithium hydroxide (66mg,2.8mmol) and the reaction mixture was stirred at room temperature for 30 min. With 2N H2SO4The reaction mixture was acidified to pH 7-8 with aqueous solution and extracted with EtOAc (20mL x 3). The combined organic layers were washed with brine and dried (MgSO)4) And concentrated to give the title compound.1H NMR(D2O,400MHz):8.10(s,1H),7.98(s,1H),7.72(s,1H),7.51(s,1H),7.40(d,J＝8.0Hz,1H),7.32(d,J＝8.4Hz,1H),4.41(s,2H),3.32(s,3H),2.13(s,3H)。
E) (R) -3- (methoxymethyl) -5- (5-methylpyridin-2-yl) -N- (1- (2-methylpyrimidin-5-yl) ethyl) benzamide
To a solution of 3- (methoxymethyl) -5- (5-methylpyridin-2-yl) benzoic acid (30mg,0.12mmol) in N, N-dimethylformamide (1.5mL) were added (R) -1- (2-methylpyrimidin-5-yl) ethylamine (40mg,0.29mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol), and N, N-diisopropylethylamine (100. mu.L, 0.57 mmol). The reaction mixture was stirred at 25 ℃ for 16 hours and then purified by preparative HPLC to give the final product.
LC-MS:377.2[M+1]+；1H NMR(400MHz,CDCl3):8.69(s,2H),8.49(d,1H,J＝1.1Hz),8.30(bs,1H),8.07(bs,1H),7.80(bs,1H),7.69(d,1H,J＝8.1Hz),7.58(dd,1H,J＝8.1,2.0Hz),6.79(d,1H,J＝7.2Hz),5.34(m,1H),4.56(s,2H),3.43(s,3H),2.72(s,3H),2.38(s,3H),1.65(d,3H,J＝7.1Hz)。
Compound 280
5- (hydroxy (thiazol-2-yl) methyl) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
A) 3-bromo-5- (hydroxymethyl) benzoic acid methyl ester
Sodium tetrahydroborate (10g,0.28mol) was slowly added to a stirred solution of dimethyl 5-bromoisophthalate (5.4g,20mmol) in methanol (300mL) at 0 ℃. The mixture was stirred at rt for 3h, then treated with water (300 mL). Removing volatile substances in vacuo, using CH2Cl2The aqueous phase was extracted (100mL x 3). The combined organic layers were washed with brine, anhydrous MgSO4Drying and vacuum concentrating. The resulting oil was purified by flash chromatography to give the title compound.
B) 3-bromo-5-formylbenzoic acid methyl ester
Methyl 3-bromo-5- (hydroxymethyl) benzoate (149mg,0.61mmol) and manganese (IV) oxide (529mg,6.08mmol) and CH2Cl2The mixture (3mL) was stirred at rt overnight. The mixture was filtered through Celite and CH was used2Cl2The filter cake is washed. The filtrate was concentrated to give the title compound.
C) 3-bromo-5- (hydroxy (thiazol-2-yl) methyl) benzoic acid methyl ester
To a stirred solution of 2-bromothiazole (0.40g,24.4mmol) in THF (60mL) at rt under a nitrogen atmosphere was added a 2M solution of isopropylmagnesium chloride in THF (10mL,20 mmol). The mixture was stirred at rt for 1h, then cooled to 0 ℃. Methyl 3-bromo-5-formylbenzoate (1.0g,4.1mmol) was added and the reaction mixture was stirred at 0 ℃ for 30 minutes. The reaction mixture was quenched with water (30mL) and CH2Cl2(60mL x 3). With anhydrous MgSO4The combined organic layers were dried and concentrated. The residue was purified by flash chromatography to give the title compound (0.90 g).
D)5- (hydroxy (thiazol-2-yl) methyl) -4' -methylbiphenyl-3-carboxylic acid methyl ester
To methyl 3-bromo-5- (hydroxy (thiazol-2-yl) methyl) benzoate (320mg,0.98mmol), p-tolylboronic acid (0.16g,1.2 m) in an argon atmospheremol), potassium carbonate (0.20g,1.5mmol), 1, 4-bisTo a mixture of alkane (5mL) and water (0.5mL,0.03mol) was added tetrakis (triphenylphosphine) palladium (0) (60mg,0.052 mmol). The reaction mixture was heated at 95 ℃ for 8 h. After cooling, the mixture was treated with water (15mL) and extracted with EtOAc (20mL × 3). The combined organic layers were dried (MgSO)4) And (4) concentrating. The residue was purified by flash chromatography to give the title compound.
E)5- (hydroxy (thiazol-2-yl) methyl) -4' -methylbiphenyl-3-carboxylic acid
To a solution of methyl 5- (hydroxy (thiazol-2-yl) methyl) -4' -methylbiphenyl-3-carboxylate (170mg,0.50mmol) in 1, 4-bisTo a stirred solution of alkane (1.5mL) and water (1.5mL) was added lithium hydroxide (42mg,1.8 mmol). After stirring for 4h at rt, with NH4The reaction mixture was worked up with aqueous Cl and extracted with EtOAc (15mL × 3). With anhydrous MgSO4The combined organic layers were dried and purified by silica gel column to give the title compound.1H NMR(CDCl3,400MHz):8.19(s,1H),8.16(s,1H),7.86(s,1H),7.72(d,J＝2.4Hz,1H),7.41(d,J＝7.6Hz,2H),7.24(m,1H),7.14(d,J＝7.6Hz,2H),6.21(s,1H),5.28(s,1H),2.32(s,3H)。
F)5- (hydroxy (thiazol-2-yl) methyl) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a stirred solution of 5- (hydroxy (thiazol-2-yl) methyl) -4' -methylbiphenyl-3-carboxylic acid (31mg,0.095mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (50mg,0.24mmol), N, N, N ', N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol), and N, N-diisopropylethylamine (300. mu.L, 1.72 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the final product as a light brown solid.
LC-MS:444.1[M+1]+；1H NMR(400MHz,DMSO-d6):8.96(d,1H,J＝7.8Hz),8.47(d,1H,J＝2.0Hz),8.05(bs,1H),7.90(bs,1H),7.85(bs,1H),7.72-7.58(m,5H),7.31(d,2H,J＝8.0Hz),7.21(d,1H,J＝8.0Hz),6.95(d,1H,J＝3.7Hz),6.08(d,1H,J＝4.2Hz),5.18(m,1H),2.43(s,3H),2.36(s,3H),1.51(d,3H,J＝7.0Hz)。
Compound 290
(R) -2 '-cyano-5- (hydroxymethyl) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
A)2 '-cyano-5- (hydroxymethyl) -4' -methylbiphenyl-3-carboxylic acid methyl ester
To a solution of dimethyl 2 '-cyano-4' -methylbiphenyl-3, 5-dicarboxylate (343mg,1.05mmol) in methanol (20mL) was slowly added sodium tetrahydroborate (0.60g,16mmol) at 0 ℃. The mixture was stirred at rt for 6h, then treated with water (20 mL). Removing volatile substances in vacuo, using CH2Cl2The aqueous phase was extracted (30mL x 3). The combined organic layers were washed with brine, anhydrous MgSO4Drying and vacuum concentrating. The residue was purified by flash chromatography to give the title compound.
B)2 '-cyano-5- (hydroxymethyl) -4' -methylbiphenyl-3-carboxylic acid
To a solution of methyl 2 '-cyano-5- (hydroxymethyl) -4' -methylbiphenyl-3-carboxylate (220mg,0.78mmol) in 1, 4-bisTo a stirred solution of alkane (5mL) and water (1mL) was added lithium hydroxide (60mg,2.5 mmol). The reaction mixture was stirred at rt for 8h, then with NH4Aqueous Cl and EtOAc. Separating the organic layer with anhydrous MgSO4Drying and concentrating. The residue was purified by silica gel chromatography to give the title compound.1H NMR (acetone-d 6,400mhz) 8.16(m,1H),8.11(m,1H),7.81(m,1H),7.73(m,1H),7.65-7.61(m,1H),7.56(d,1H, J ═ 8.0Hz),4.81(s,2H),2.46(s, 3H).
C) (R) -2 '-cyano-5- (hydroxymethyl) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a stirred solution of 2 '-cyano-5- (hydroxymethyl) -4' -methylbiphenyl-3-carboxylic acid (40mg,0.15mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (400. mu.L, 2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the final product as a pale solid.
LC-MS:386.4[M+1]+；1H NMR(400MHz,DMSO-d6):8.93(d,1H,J＝7.8Hz),8.47(d,1H,J＝2.2Hz),7.93(bs,1H),7.91(bs,1H),7.81(bs,1H),7.70-7.60(m,3H),7.57(d,1H,J＝8.0Hz),7.20(d,1H,J＝8.0Hz),5.41(t,1H,J＝5.7Hz),5.18(m,1H),4.62(d,2H,J＝5.7Hz),2.43(s,3H),2.41(s,3H),1.50(d,3H,J＝7.1Hz)。
Compound 295
2 '-cyano-4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- (2,2, 2-trifluoro-1-hydroxyethyl) biphenyl-3-carboxamide
A) 3-bromo-5- (2,2, 2-trifluoro-1-hydroxyethyl) benzoic acid methyl ester
To a stirred mixture of methyl 3-bromo-5-formylbenzoate (0.60g,2.5mmol), tetra-N-butylammonium bromide (0.90g,2.8mol), potassium fluoride (10mg,0.17mmol) and toluene (10mL) was added (trifluoromethyl) trimethylsilane (0.74mL,5.0mmol) at-20 ℃. The reaction mixture was stirred for 20min and then quenched with water. To the direction ofTo the mixture were added 1M aqueous HCl (2mL) and 1, 4-bis Alkane (12mL) and the mixture was stirred for 30 min. The aqueous phase was extracted with EtOAc (3 × 20 mL). The combined organic layers were washed with brine, anhydrous MgSO4Drying, and concentrating under reduced pressure. The residue was purified by flash column to give the title compound.
B)2 '-cyano-4' -methyl-5- (2,2, 2-trifluoro-1-hydroxyethyl) biphenyl-3-carboxylic acid methyl ester
A mixture of methyl 3-bromo-5- (2,2, 2-trifluoro-1-hydroxyethyl) benzoate (300 mg,0.96mmol), 5-methyl-2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzonitrile (280mg,1.1mmol), tetrakis (triphenylphosphine) palladium (0) (60mg,0.052mmol) in N, N-dimethylformamide (0.75mL) and toluene (1.5mL) was subjected to microwave irradiation at 110 ℃ for 1h under a nitrogen atmosphere. After cooling, the mixture was diluted with water (15mL) and extracted with EtOAc (20mL × 3). The combined organic layers were washed with brine, anhydrous MgSO4Drying and concentrating. The residue was purified by flash chromatography to give the title compound.
C)2 '-cyano-4' -methyl-5- (2,2, 2-trifluoro-1-hydroxyethyl) biphenyl-3-carboxylic acid
To a solution of methyl 2 '-cyano-4' -methyl-5- (2,2, 2-trifluoro-1-hydroxyethyl) biphenyl-3-carboxylate (250mg,0.72mmol) in 1, 4-bisTo a stirred solution of alkane (5mL) and water (2.5mL) was added lithium hydroxide (50mg,2.1 mmol). The reaction mixture was stirred at rt for 4h, then with NH4Aqueous Cl and EtOAc. Separating the organic layer with anhydrous MgSO4Drying and concentrating. The residue was purified by silica gel chromatography to give the title compound.1H NMR(CDCl3400MHz) 8.27(s,1H),8.24(s,1H),7.95(s,1H),7.58(s,1H),7.48 and 7.43(AB, J ═ 8.0Hz,2H),5.17(q, J ═ 6.4Hz,1H),2.43(s, 3H).
D)2 '-cyano-4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- (2,2, 2-trifluoro-1-hydroxyethyl) biphenyl-3-carboxamide
To a solution of 2' -cyano-4 ' -methyl-5- (2,2, 2-trifluoro-1-hydroxyethyl) biphenyl-3-carboxylic acid (32mg,0.095mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (50mg,0.24mmol), N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol), and N, N-diisopropylethylamine (300 μ L,1.72 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and purified by preparative HPLC to give the final product as a pale solid.
LC-MS:454.3[M+1]+；1H NMR(400MHz,DMSO-d6):9.00(d,1H,J＝7.7Hz),8.48(d,1H,J＝2.1Hz),8.09(s,2H),7.83(s,2H),7.70-7.55(m,3H),7.21(d,1H,J＝8.0Hz),7.07(d,1H,J＝5.6Hz),5.35(m,1H),5.19(m,1H),2.43(s,3H),2.42(s,3H),1.51(d,3H,J＝7.1Hz)。
Compound 297
2 '-cyano-5- (1, 2-dihydroxyethyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
A) 3-bromo-5-vinylbenzoic acid methyl ester
Methyl triphenyl phosphonium bromide is reacted at-78 ℃ under nitrogen atmosphere(350mg,0.97mmol) to a stirred solution in THF (3mL) was added dropwise a 2.5M solution of n-butyllithium in tetrahydrofuran (0.31mL,0.78 mmol). The mixture was gradually warmed until the yellow color continued to appear. The mixture was cooled to 0 ℃ and a solution of methyl 3-bromo-5-formylbenzoate (157mg,0.65mmol) in THF (2mL) was added dropwise. Stirring at 0 deg.C for 20min, adding saturated NH4The mixture was quenched with aqueous Cl and extracted with EtOAc.The organic layer was separated, washed with brine, dried and concentrated. The residue was purified by flash column to give the title compound.
B) 3-bromo-5- (1, 2-dihydroxyethyl) benzoic acid methyl ester
To a stirred solution of methyl 3-bromo-5-vinylbenzoate (1.6g,6.6mmol) and N-methylmorpholine N-oxide (2.0g,17.1mmol) in acetone (40mL) and water (10mL) was added dropwise a 5% aqueous solution of osmium tetroxide (0.50g,0.10 mmol). The reaction mixture was stirred at rt for 3h, then saturated Na was added2S2O3An aqueous solution. After stirring for 30min, the reaction mixture was extracted with EtOAc (100mL × 3). The combined organic layers were washed with water and brine, dried and concentrated. The residue was purified by flash column to give the title compound.
C)2 '-cyano-5- (1, 2-dihydroxyethyl) -4' -methylbiphenyl-3-carboxylic acid methyl ester
A mixture of methyl 3-bromo-5- (1, 2-dihydroxyethyl) benzoate (100mg,0.36mmol), 5-methyl-2- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) benzonitrile (130mg,0.54mmol), tetrakis (triphenylphosphine) palladium (0) (40mg,0.035mmol), potassium carbonate (75mg,0.54mmol), N-dimethylformamide (0.5mL) and toluene (1mL) was subjected to microwave irradiation at 110 ℃ for 1h under a nitrogen atmosphere. After cooling, the mixture was diluted with water (5mL) and extracted with EtOAc (15mL x 3). The combined organic layers were washed with brine, anhydrous MgSO4Drying and vacuum concentrating. The residue was purified by flash column to give the title compound.
D)2 '-cyano-5- (1, 2-dihydroxyethyl) -4' -methylbiphenyl-3-carboxylic acid
To a solution of methyl 2 '-cyano-5- (1, 2-dihydroxyethyl) -4' -methylbiphenyl-3-carboxylate (140mg,0.45mmol) in 1, 4-bisTo a stirred solution of alkane (5mL) and water (2mL) was added lithium hydroxide (40mg,1.67 mmol). The reaction mixture was stirred at rt for 6h, then with NH4Aqueous Cl, extracted with EtOAc (3 ×).With anhydrous MgSO4The combined organic layers were dried and concentrated to give the title compound.1H NMR (400MHz, acetone-d 6):8.21(m,1H),8.13(m,1H),7.87(m,1H),7.73(s,1H),7.66-7.61(m,1H),7.58(d,1H, J ═ 7.6Hz),4.91(dd,1H, J ═ 7.2,4.4Hz),3.75(dd,1H, J ═ 11.2,4.4Hz),3.65(dd,1H, J ═ 11.2,7.2Hz),2.47(s, 3H).
E)2 '-cyano-5- (1, 2-dihydroxyethyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
To a solution of 2' -cyano-5- (1, 2-dihydroxyethyl) -4' -methylbiphenyl-3-carboxylic acid (30mg,0.10mmol) in N, N-dimethylformamide (1mL) were added (6-methylpyridin-3-yl) methylamine (35mg,0.26mmol), N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol), and N, N-diisopropylethylamine (100 μ L,0.57 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the final product as a pale solid.
LC-MS:402.1[M+1]+；1H NMR(400MHz,DMSO-d6):9.14(m,1H),8.47(bs,1H),7.94(m,2H),7.79(bs,1H),7.70-7.50(m,4H),7.20(m,1H),5.46(bs,1H),4,80(bs,1H),4.66(bs,1H),4.46(bs,2H),3.49(bs,2H),2.43(s,3H),2.40(s,3H)。
Compound 308
4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- (tetrahydrofuran-3-yloxy) biphenyl-3-carboxamide
A)4' -methyl-5- (tetrahydrofuran-3-yloxy) biphenyl-3-carboxylic acid methyl ester
5-hydroxy-4' -methylbiphenyl-3-carboxylic acid methyl ester (120mg,0.47mmol), triphenylphosphine (120mg,0.47mmol), 3-hydroxytetrahydrofuran (45mg,0.50mmol) in CH at rt2Cl2(7mL) to the stirred solution was slowly added azobisdicarboxylic acidIsopropyl ester (100mg,0.50mmol) in CH2Cl2(2 mL). The mixture was stirred at rt overnight and then concentrated in vacuo. The residue was purified by flash chromatography to give the title compound.
B)4' -methyl-5- (tetrahydrofuran-3-yloxy) biphenyl-3-carboxylic acid
To a solution of methyl 4' -methyl-5- (tetrahydrofuran-3-yloxy) biphenyl-3-carboxylate (150mg,0.46mmol) in 1, 4-bisTo a stirred solution of alkane (5mL) and water (0.5mL) was added lithium hydroxide (55mg,2.3 mmol). The reaction mixture was stirred at room temperature for 2 hours, then 2NH was used2SO4The aqueous solution was acidified to pH4-5 and concentrated in vacuo. Treating the residue with water, using CH2Cl2(10 mL. times. 3). The combined organic layers were dried (MgSO)4) And concentrated to give the title compound.1H NMR(CD3OD,400MHz):7.85(t,J＝1.6Hz,1H),7.52(d,2H,J＝8.4Hz),7.48(dd,1H＝2.4,1.2Hz),7.34(dd,1H,J＝2.4,1.6Hz),7.28(d,2H,J＝8.4Hz),5.15(m,1H),4.05-3.87(m,4H),2.38(s,3H),2.36-2.27(m,1H),2.20-2.11(m,1H)。
C)4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- (tetrahydrofuran-3-yloxy) biphenyl-3-carboxamide
To a solution of 4 '-methyl-5- (tetrahydrofuran-3-yloxy) biphenyl-3-carboxylic acid (40mg,0.13mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (400 μ L,2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and purified by preparative HPLC to give the final product as a pale solid.
LC-MS:417.1[M+1]+；1H NMR(400MHz,DMSO-d6):8.90(d,1H,J＝7.7Hz),8.47(d,1H,J＝2.2Hz),7.73(bs,1H),7.70-7.60(m,3H),7.35(bs,1H),7.32-7.25(m,3H),7.21(d,1H,J＝8.0Hz),5.20(m,2H),4.00-3.70(m,4H),2.43(s,3H),2.35(s,3H),2.25(m,1H),2.00(m,1H),1.51(d,3H,J＝7.1Hz)。
Compound 310
4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- ((tetrahydrofuran-2-yl) methoxy) biphenyl-3-carboxamide
A)4' -methyl-5- ((tetrahydrofuran-2-yl) methoxy) biphenyl-3-carboxylic acid methyl ester
To methyl 5-hydroxy-4' -methylbiphenyl-3-carboxylate (150mg,0.59mmol), triphenylphosphine (150mg,0.59mmol), tetrahydro-2-furancarbinol (60mg,0.59mmol) in CH at rt2Cl2(8mL) to a stirred solution was slowly added diisopropyl azodicarboxylate (200mg,1.0mmol) in CH2Cl2(3 mL). The mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by flash chromatography to give the title compound.
B)4' -methyl-5- ((tetrahydrofuran-2-yl) methoxy) biphenyl-3-carboxylic acid
To a solution of methyl 4' -methyl-5- ((tetrahydrofuran-2-yl) methoxy) biphenyl-3-carboxylate (130mg,0.40mmol) in 1, 4-bisTo a stirred solution of alkane (5mL) and water (0.5mL) was added lithium hydroxide (50mg,2.1 mmol). The reaction mixture was stirred at room temperature for 2 hours and then with 2N H2SO4The aqueous solution was acidified to pH4-5 and concentrated in vacuo. Treating the residue with water, using CH2Cl2(10 mL. times. 3). The combined organic layers were dried (MgSO)4) And concentrated to give the title compound.1H NMR(CD3OD,400MHz):7.84(t,J＝1.6Hz,1H),7.54-7.50(m,3H),7.38(dd,1H,J＝2.8,1.6Hz),7.27(d,2H,J＝8.0Hz),4.34-4.27(m,1H),4.12(dd,1H,J＝10.0,3.6Hz),4.04(dd,1H,J＝10.0,6.4Hz),3.97-3.80(m,2H),2.38(s,3H),2.20-1.80(m,4H)。
4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- ((tetrahydrofuran-2-yl) methoxy) biphenyl-3-carboxamide
To a stirred solution of 4 '-methyl-5- ((tetrahydrofuran-2-yl) methoxy) biphenyl-3-carboxylic acid (40mg,0.13mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (400 μ L,2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the final product as a light brown solid.
LC-MS:430.9[M+1]+；1H NMR(400MHz,DMSO-d6):8.90(d,1H,J＝7.7Hz),8.47(d,1H,J＝2.2Hz),7.71(bs,1H),7.70-7.60(m,3H), 7.38(bs,1H),7.33(t,1H,J＝1.6Hz),7.29(d,2H,J＝8.0Hz),7.21(d,1H,J＝8.0Hz),5.18(m,1H),4.19(m,1H),4.15-3.95(m,2H),3.79(m,1H),3.67(m,1H),2.43(s,3H),2.35(s,3H),2.02(m,1H),1.85(m,2H),1.70(m,1H),1.51(d,3H,J＝7.1Hz)。
Compound 315
2 '-cyano-5- (2, 3-dihydroxypropoxy) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
A)2 '-cyano-5- ((2, 2-dimethyl-1, 3-dioxolan-4-yl) methoxy) -4' -methylbiphenyl-3-carboxylic acid methyl ester
To methyl 2 '-cyano-5-hydroxy-4' -methylbiphenyl-3-carboxylate (150mg,0.56mmol), triphenylphosphine (220mg,0.84mmol), 2-dimethyl-1, 3-dioxolane-4-methanol (82mg,0.62mmol) in CH at room temperature2Cl2(5mL) in a stirred solutionDiisopropyl azodicarboxylate (340mg,1.7mmol) was slowly added to CH2Cl2(3 mL). The reaction mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by flash chromatography to give the title compound.
B)2 '-cyano-5- ((2, 2-dimethyl-1, 3-dioxolan-4-yl) methoxy) -4' -methylbiphenyl-3-carboxylic acid
To a solution of methyl 2 '-cyano-5- ((2, 2-dimethyl-1, 3-dioxolan-4-yl) methoxy) -4' -methylbiphenyl-3-carboxylate (72mg,0.19mmol) in tetrahydrofuran (3mL) was added 2.5M aqueous lithium hydroxide solution (1mL,2.5 mmol). The reaction mixture was stirred at 60 ℃ overnight. The aqueous solution was acidified to pH 5 with 15% HCl (aq) and extracted with EtOAc. The combined organic layers were concentrated in vacuo to afford the title compound.
C)2 '-cyano-5- (2, 3-dihydroxypropoxy) -4' -methylbiphenyl-3-carboxylic acid
To a stirred solution of 2 '-cyano-5- ((2, 2-dimethyl-1, 3-dioxolan-4-yl) methoxy) -4' -methyldibenzene-3-carboxylic acid (63mg,0.17mmol) in THF (3mL) was added 1MHCl aqueous solution (3mL,3 mmol). The reaction mixture was stirred at room temperature for 4 hours, then concentrated in vacuo. The residue was treated with water and extracted with EtOAc. With anhydrous MgSO4The combined organic layers were dried and concentrated to give the title compound.1H NMR(CD3OD,400MHz):7.77(t,1H,J＝1.6Hz),7.67(m,2H),7.58(dd,1H,J＝8.0,2.0Hz),7.51(d,1H,J＝8.0Hz),7.38(t,1H,J＝2.0Hz),4.19(dd,1H,J＝9.6,4.0Hz),4.10(dd,1H,J＝9.6,6.0Hz),4.02(m,1H),3.72(dd,1H,J＝11.2,5.6Hz),3.67(dd,1H,J＝11.2,5.6Hz),2.45(s,3H)。
D)2 '-cyano-5- (2, 3-dihydroxypropoxy) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a solution of 2' -cyano-5- (2, 3-dihydroxypropoxy) -4' -methylbiphenyl-3-carboxylic acid (30mg,0.092mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol), and N, N-diisopropylethylamine (400 μ L,2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and purified by preparative HPLC to give the final product as a pale solid.
LC-MS:446.6[M+1]+；1H NMR(400MHz,DMSO-d6):8.92(d,1H,J＝7.4Hz),8.46(bs,1H),7.79(bs,1H),7.70-7.55(m,4H),7.53(bs,1H),7.27(bs,1H),7.20(d,1H,J＝7.8Hz),5.17(m,1H),5.00(m,1H),4.70(t,1H,J＝3.8Hz),4.11(m,1H),3.97(m,1H),3.83(m,1H),3.46(t,2H,J＝4.8Hz),2.43(s,3H),2.41(s,3H),1.49(d,3H,J＝6.2Hz)。
Compound 324
2 '-cyano-4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- (morpholin-2-ylmethoxy) biphenyl-3-carboxamide
A)2- ((2 '-cyano-5- (methoxycarbonyl) -4' -methylbiphenyl-3-yloxy) methyl) morpholine-4-carboxylic acid tert-butyl ester
To methyl 2 '-cyano-5-hydroxy-4' -methylbiphenyl-3-carboxylate (150mg,0.56mmol), triphenylphosphine (220mg,0.84mmol), tert-butyl 2- (hydroxymethyl) morpholine-4-carboxylate (130mg,0.62mmol) (bioorg.med.chem.lett.2007,17,533) in CH at room temperature under a nitrogen atmosphere2Cl2(5mL) to a stirred solution was slowly added diisopropyl azodicarboxylate (340mg,1.7mmol) in CH2Cl2(3 mL). The mixture was stirred at room temperature overnight and then concentrated in vacuo. The residue was purified by flash chromatography to give the title compound.
B)5- ((4- (tert-Butoxycarbonyl) morpholin-2-yl) methoxy) -2 '-cyano-4' -methylbiphenyl-3-carboxylic acid
To tert-butyl 2- ((2 '-cyano-5- (methoxycarbonyl) -4' -methylbiphenyl-3-yloxy) methyl) morpholine-4-carboxylate (10)7mg,0.23mmol) in THF (2mL) was added 2.5M aqueous lithium hydroxide (0.52mL,1.3 mmol). The reaction mixture was stirred at 60 ℃ overnight, then acidified to pH 5 with 15% HCl (aq) and extracted with EtOAc. The combined organic layers were concentrated in vacuo to afford the title compound.1H NMR(CDCl3,400MHz):7.86(t,1H,J＝1.6Hz),7.70(t,1H,J＝1.6Hz),7.59(s,1H),7.47(dd,1H,J＝8.0,1.2Hz),7.42(d,1H,J＝8.0Hz),7.37(s,1H),4.20-3.56(m,7H),3.10-2.90(m,2H),2.45(s,3H),1.49(s,9H)。
C)2 '-cyano-4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) -5- (morpholin-2-ylmethoxy) biphenyl-3-carboxamide
To a solution of 5- ((4- (tert-butoxycarbonyl) morpholin-2-yl) methoxy) -2' -cyano-4 ' -methylbiphenyl-3-carboxylic acid (50mg,0.11mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N ' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (400 μ L,2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and purified by preparative HPLC to give the Boc-protected product, which was dissolved in CH2Cl2(5mL), trifluoroacetic acid (1mL) was added. The mixture was stirred at rt overnight and LC-MS showed deprotection complete. The mixture was concentrated in vacuo and the residue was purified by preparative HPLC to give the final product as a white solid.
LC-MS:471.6[M+1]+；1H NMR(400MHz,DMSO-d6):8.90(d,1H,J＝7.7Hz),8.46(d,1H,J＝2.1Hz),8.00(bs,1H),7.70-7.55(m,4H), 7.53(bs,1H),7.29(bs,1H),7.21(d,1H,J＝8.0Hz),5.17(m,1H),4.05(m,2H),3.75(m,2H),3.49(m,1H),2.92(m,1H),2.65(m,2H),2.53(m,1H),2.43(s,3H),2.41(s,3H),1.50(d,3H,J＝7.1Hz)。
Compound 336
5- (4-hydroxytetrahydrofuran-3-yloxy) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
A)5- (4-Hydroxytetrahydrofuran-3-yloxy) -4' -methylbiphenyl-3-carboxylic acid methyl ester
Methyl 5-hydroxy-4' -methylbiphenyl-3-carboxylate (370mg,1.4mmol), potassium carbonate (421mg,3.05mmol), DMSO (8mL), and 3, 6-dioxabicyclo [3.1.0 ]]A stirred mixture of hexane (260mg,3.0mmol) was heated at 110 deg.C overnight. After cooling, the reaction mixture was diluted with EtOAc. With NaHCO3The organic phase was washed with aqueous (saturated) solution and anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by flash chromatography to give the title compound.
B)5- (4-hydroxytetrahydrofuran-3-yloxy) -4' -methylbiphenyl-3-carboxylic acid
To a solution of methyl 5- (4-hydroxytetrahydrofuran-3-yloxy) -4' -methylbiphenyl-3-carboxylate (140mg,0.40mmol) in 1, 4-bisTo a stirred solution of alkane (2.8mL) and water (2.8mL) was added lithium hydroxide (50mg,2.1 mmol). The reaction mixture was stirred at room temperature for 2 hours and then with 2N H2SO4The aqueous solution was acidified to pH4-5 and concentrated in vacuo. The residue was treated with water and extracted with EtOAc (10mL × 2). The combined organic layers were dried (MgSO)4) And concentrated to give the title compound.1H NMR(CD3OD,400MHz):7.87(t,J＝1.6Hz,1H),7.55-7.51(m,3H),7.42(t,J＝2.0Hz,1H),7.28(d,J＝7.6Hz,2H),4.84(d,J＝4.4Hz,1H),4.38(d,J＝4.0Hz,1H),4.23(dd,J＝10.4,4.4Hz,1H),4.05(dd,J＝9.6,4.4Hz,1H),3.94(dd,J＝10.4,1.6Hz,1H),3.77(dd,J＝9.6,1.6Hz,1H),2.38(s,3H)。
C)5- (4-hydroxytetrahydrofuran-3-yloxy) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a solution of 5- (4-hydroxytetrahydrofuran-3-yloxy) -4 '-methylbiphenyl-3-carboxylic acid (35mg,0.11 mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (100mg,0.26mmol), and N, N-diisopropylethylamine (400 μ L,2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and then purified by preparative HPLC to give the final product as a white solid.
LC-MS:432.8[M+1]+；1H NMR(400MHz,DMSO-d6):8.90(d,1H,J＝7.8Hz),8.47(d,1H,J＝1.6Hz),7.73(bs,1H),7.70-7.60(m,3H),7.41(bs,1H),7.38(bs,1H),7.29(d,2H,J＝7.9Hz),7.21(d,1H,J＝8.0Hz),5.53(bs,1H),5.17(m,1H),4.85(d,1H,J＝3.7Hz),4.24(bs,1H),4.08(dd,1H,J＝10.2,4.1Hz),3.93(dd,1H,J＝9.5,4.4Hz),3.80(d,1H,J＝10.2Hz),3.61(d,1H,J＝9.4Hz),2.43(s,3H),2.36(s,3H),1.50(d,3H,J＝7.0Hz)。
Compound 338
2 '-cyano-5- (4-hydroxypyrrolidin-3-yloxy) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
A)3- (2 '-cyano-5- (methoxycarbonyl) -4' -methylbiphenyl-3-yloxy) -4-hydroxypyrrolidine-1-carboxylic acid tert-butyl ester
Methyl 2 '-cyano-5-hydroxy-4' -methylbiphenyl-3-carboxylate (300mg,1.1mol), potassium carbonate (320mg,2.4mmol), DMSO (8mL) and 6-oxa-3-aza-bicyclo [ 3.1.0% ]A stirred mixture of tert-butyl hexane-3-carboxylate (2.4mmol) (J.Am.chem.Soc.2008,130,3900) was heated at 135 deg.C overnight. After cooling, the reaction mixture was diluted with EtOAc. With NaHCO3The organic phase was washed with aqueous (saturated) solution and anhydrous MgSO4Drying, filtering and concentrating. The residue was purified by flash chromatography (0-50% EtOAc/hexanes) to give the title compound.
B)5- (1- (tert-Butoxycarbonyl) -4-hydroxypyrrolidin-3-yloxy) -2 '-cyano-4' -methylbiphenyl-3-carboxylic acid
To a stirred solution of tert-butyl 3- (2 '-cyano-5- (methoxycarbonyl) -4' -methylbiphenyl-3-yloxy) -4-hydroxypyrrolidine-1-carboxylate (310mg,0.68mmol) in THF (3mL) was added 2.5M aqueous lithium hydroxide solution (1.5mL,3.8 mmol). The reaction mixture was stirred at 60 ℃ overnight. The aqueous solution was acidified to pH 5 with 15% HCl (aq) and extracted with EtOAc. The combined organic layers were concentrated in vacuo to afford the title compound.
C)2 '-cyano-5- (4-hydroxypyrrolidin-3-yloxy) -4' -methyl-N- ((R) -1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a solution of 5- (1- (tert-butoxycarbonyl) -4-hydroxypyrrolidin-3-yloxy) -2 '-cyano-4' -methylbiphenyl-3-carboxylic acid (60mg,0.14mol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (60mg,0.29mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (120mg,0.32mmol), and N, N-diisopropylethylamine (400. mu.L, 2.3 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and purified by preparative HPLC to give the Boc-protected product, which was dissolved in CH2Cl2(5mL), trifluoroacetic acid (1mL) was added. The mixture was stirred at room temperature for 2hr, then concentrated in vacuo. The residue was purified by preparative HPLC to give the final product as a pale solid.
LC-MS:457.3[M+1]+；1H NMR(400MHz,DMSO-d6):8.90(d,1H,J＝7.6Hz),8.46(bs,1H),7.80(bs,1H),7.70-7.55(m,4H),7.52(bs,1H),7.33(bs,1H),7.20(d,1H,J＝8.2Hz),5.28(bs,1H),5.16(m,1H),4.65(bs,1H),4.16(bs,1H),3.50-3.25(m,2H),3.05(dd,1H,J＝11.7,4.4Hz),2.88(d,1H,J＝12.6Hz),2.73(d,1H,J＝11.7Hz),2.43(s,3H),2.41(s,3H),1.49(d,3H,J＝6.9Hz)。
Compound 340
(R) -5- ((dimethylamino) methyl) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
A) 3-bromo-5- ((dimethylamino) methyl) benzoic acid methyl ester
A mixture of methyl 3-bromo-5-formylbenzoate (200mg,0.82mmol), methanol (3mL),2.0M dimethylamine in THF (0.3mL,6mmol), zinc dichloride (30mg,0.22mmol) and sodium cyanoborohydride (200mg,3.2mmol) was stirred at 0 deg.C for 1 h. The mixture was diluted with water (10mL) and the aqueous layer was extracted with EtOAc (15mL x 3). With anhydrous MgSO4The combined organic layers were dried and concentrated. The residue was purified by silica gel chromatography to give the title compound.
B)5- ((dimethylamino) methyl) -4' -methylbiphenyl-3-carboxylic acid ester
A mixture of methyl 3-bromo-5- ((dimethylamino) methyl) benzoate (50mg,0.18mmol), p-tolylboronic acid (30mg,0.22mmol), tetrakis (triphenylphosphine) -palladium (0) (10mg,0.009mmol), N-dimethylformamide (0.5mL), and toluene (1.5mL) was subjected to microwave irradiation at 110 ℃ for 1h under a nitrogen atmosphere. After cooling, the mixture was diluted with water (15mL) and extracted with EtOAc (15mL x 3). With anhydrous MgSO4The combined organic layers were dried and concentrated in vacuo. The residue was purified by flash chromatography to give the title compound.
C)5- ((dimethylamino) methyl) -4' -methylbiphenyl-3-carboxylic acid
To a solution of methyl 5- ((dimethylamino) methyl) -4' -methylbiphenyl-3-carboxylate (90mg,0.32mmol) in 1, 4-bisTo a stirred solution of alkane (1.5mL) and water (1.5mL) was added lithium hydroxide (20mg,0.83 mmol). The reaction mixture was stirred at room temperature for 2h, then NH was used4Aqueous Cl, extracted with EtOAc (15mL × 3). With anhydrous MgSO4The combined organic layers were dried and concentrated. By chromatography on silica gelThe residue was purified by the method to give the title compound.1HNMR(CD3OD,400MHz):8.29(s,1H),7.79(s,1H),7.60(s,1H),7.59(d,J＝8.0Hz,2H),7.28(d,J＝8.0Hz,2H),4.27(s,2H),2.81(s,6H),2.38(s,3H)。
D) (R) -5- ((dimethylamino) methyl) -4' -methyl-N- (1- (6-methylpyridin-3-yl) ethyl) biphenyl-3-carboxamide
To a solution of 5- ((dimethylamino) methyl) -4 '-methylbiphenyl-3-carboxylic acid (20mg,0.074mmol) in N, N-dimethylformamide (1mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (40mg,0.19mmol), N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (60mg,0.16mmol), and N, N-diisopropylethylamine (200 μ L,1.15 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and purified by preparative HPLC to give the final product.
LC-MS:388.3[M+1]+；1H NMR(400MHz,DMSO-d6):8.93(d,1H,J＝7.8Hz),8.48(d,1H,J＝2.2Hz),8.01(t,1H,J＝1.4Hz),7.75(bs,1H),7.70-7.65(m,2H),7.62(d,2H,J＝8.1Hz),7.31(d,2H,J＝8.0Hz),7.21(d,1H,J＝8.0Hz),5.19(m,1H),3.48(s,2H),2.43(s,3H),2.36(s,3H),2.17(s,6H),1.52(d,3H,J＝7.1Hz)。
Compound 342
3- (2- (diethylamino) -1-hydroxyethyl) -5- (5-methylpyridin-2-yl) -N- ((R) -1- (6-methylpyridin-3-yl) ethyl) benzamide
A) 3-bromo-5- (oxiran-2-yl) benzoic acid methyl ester
To methyl 3-bromo-5-vinylbenzoate (1.00g,4.15mmol) in CH at 0 ℃ over a 10 minute period2Cl2(40mL) was added m-chloroperbenzoic acid (70% pure, 1.43g,5.80mmol) in portions to the stirred solution. The mixture was warmed to room temperature and stirred for 2 hoursThen it was poured onto saturated sodium bicarbonate (250 mL). The mixture was extracted with ethyl acetate (3 × 100mL), Na2SO4The combined extracts were dried and concentrated. The residue was purified by column chromatography (0-50% EtOAc/hexanes) to give the title compound as a yellow oil.1H NMR(400MHz,DMSO-d6):7.92(t,1H,J＝1.7Hz),7.86(t,1H,J＝1.5Hz),7.78(t,1H,J＝1.7Hz),4.09(q,1H,J＝2.5Hz),3.87(s,3H), 3.17-3.14(m,1H),2.92-2.90(m,1H)。
B) 3-bromo-5- (1- (diethylamino) -2-hydroxyethyl) benzoic acid methyl ester
Combine methyl 3-bromo-5- (oxiran-2-yl) benzoate (0.10g,0.39mmol), ethanol (10mL), and diethylamine (120. mu.L, 1.2mmol) in a 20mL reaction vessel. The mixture was heated at 50 ℃ overnight and then concentrated. By preparative HPLC (100X21.2mmC18 column, CH)3CN/Water [10mM Et2NH]) The residue was purified to give the title compound.
LC-MS:331.7[M+1]+；1H NMR(400MHz,DMSO-d6):7.95(t,1H,J＝1.7Hz),7.91(t,1H,J＝1.7Hz),7.80(t,1H,J＝1.7Hz),5.28(brs,1H),4.66(t,1H,J＝6.5Hz),3.86(s,3H),2.57-2.51(m,2H),2.48-2.43(m,4H),0.86(t,6H,J＝7.0Hz)。
C)3- (2- (diethylamino) -1-hydroxyethyl) -5- (5-methylpyridin-2-yl) benzoic acid methyl ester
A mixture of methyl 3-bromo-5- (2- (diethylamino) -1-hydroxyethyl) benzoate (80mg,0.24mmol), 5-methyl-2- (tributylstannyl) pyridine (100. mu.L, 0.30mmol), tetrakis (triphenylphosphine) -palladium (0) (14mg,0.012mmol), and toluene (2.6mL) was subjected to microwave irradiation at 120 ℃ for 2 hours under an argon atmosphere. The mixture was cooled to room temperature, poured into brine (20mL) and extracted with ethyl acetate (3 × 20 mL). With Na2SO4The combined extracts were dried and concentrated in vacuo. The residue was purified by column chromatography (0-100% EtOAc/hexanes) to give the title compound as a yellow oil. LC-MS 343.7[ M +1 ]]+。
D)3- (2- (diethylamino) -1-hydroxyethyl) -5- (5-methylpyridin-2-yl) benzoic acid
In a 20mL reaction vessel, combine methyl 3- (2- (diethylamino) -1-hydroxyethyl) -5- (5-methylpyridin-2-yl) benzoate (50mg,0.15mmol), tetrahydrofuran (10mL), and lithium hydroxide (8.7mg,0.36 mmol). The mixture was heated at 50 ℃ for 3 hours. After cooling, the mixture was treated with 7M aqueous HCl (52. mu.L, 0.36mmol) and concentrated to give the crude compound, which was used directly in the next step.
E)3- (2- (diethylamino) -1-hydroxyethyl) -5- (5-methylpyridin-2-yl) -N- ((R) -1- (6-methylpyridin-3-yl) ethyl) benzamide
To a solution of 3- (2- (diethylamino) -1-hydroxyethyl) -5- (5-methylpyridin-2-yl) benzoic acid (30mg,0.075mmol) in N, N-dimethylformamide (5mL) were added (R) -1- (6-methylpyridin-3-yl) ethylamine dihydrochloride (100mg,0.48mmol), N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (160mg,0.42mmol), and N, N-diisopropylethylamine (300. mu.L, 1.7 mmol). The reaction mixture was stirred at 25 ℃ for 16 h and passed through preparative HPLC (100X21.2mm C18 column, CH3CN/Water [10mM Et2NH]) Purification gave the title product.
LC-MS:447.6[M+1]+；1H NMR(400MHz,DMSO-d6):8.96(d,1H,J＝8.0Hz),8.54(bs,1H),8.49(bs,1H),8.38(bs,1H),8.19(bs,1H),7.93(d,1H,J＝8.1Hz),7.87(bs,1H),7.77-7.65(m,2H),7.22(d,1H,J＝8.1Hz),5.21(m,1H),5.06(bs,1H),4.72(bs,1H),3.40-3.30(m,2H),2.65-2.50(m,4H),2.43(s,3H),2.35(s,3H),1.53(d,3H,J＝7.0Hz),0.92(t,6H,J＝7.1Hz)。
Compound 365
5- (1-hydroxy-2-morpholinoethyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
A) 3-bromo-5-formylbenzoic acid
In a round bottom flask 3-formylbenzoic acid (10.0g,66.6mmol) and sulfuric acid (653g,6.66mol) were combined. N-bromosuccinimide (14.23g,79.9mmol) was added portionwise over a 10 minute time period, the reaction was stirred at room temperature for 3h, and the mixture was poured onto ice. The white precipitate formed was filtered, washed with cold water (5x100mL), and recrystallized from water-ethanol to give the title compound as a white solid (12.98g, 76.6%). LC-MS 227.0[ M-1 ]]-；1H NMR(400MHz,DMSO-d6):10.05(s,1H),8.40(t,1H,J＝1.5Hz),8.3(d,2H,J＝1.5Hz)。
B) 5-formyl-4' -methylbiphenyl-3-carboxylic acid
To a mixture of 3-bromo-5-formylbenzoic acid (8.0g,34.9mmol), p-tolylboronic acid (9.5g,70mmol), toluene (300mL), cesium carbonate (28g,87mmol), and water (25mL) was added tetrakis (triphenylphosphine) palladium (0) (2.0g,1.7mmol) under a nitrogen atmosphere. The mixture was heated at reflux for 5 h. After cooling, the mixture was filtered through Celite and the filter cake was washed with EtOAc. The filtrate was washed with brine, dried and concentrated. The residue was purified by column chromatography using a dichloromethane: methanol gradient (0-5%) to give the title compound (4.8g, 51%). LC-MS 239.0[ M-1 ]]-。
C) 5-formyl-4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
In a round-bottom flask was combined 5-formyl-4' -methylbiphenyl-3-carboxylic acid (3.00g,12.5mmol), (6-methylpyridin-3-yl) methylamine (1.91g,15.6mmol), N-diisopropylethylamine (6.46g,49.9mmol) and N, N-dimethylformamide (97 mL). N, N, N ', N' -tetramethyl-O- (7-azabenzotriazol-1-yl) urea hexafluorophosphate (9.50g,25.0mmol) was added in one portion and the mixture was heated at 60 ℃ for 2 h. After cooling, the mixture was poured onto saturated sodium bicarbonate (200 mL). Extracted with ethyl acetate (3 × 100 mL). The combined extracts were dried over sodium sulfate and concentrated in vacuo. The residue was purified by column chromatography using a dichloromethane methanol gradient (0-10%) to give the title compound A compound (I) is provided. LC-MS 346.2[ M +1 ]]+；1HNMR(400MHz,DMSO-d6):10.14(s,1H),8.46-8.43(m,2H),8.37-8.33(m,2H),7.72(d,2H,J＝8.0Hz),7.64(dd,1H,J＝8.0Hz),7.34(d,2H,J＝7.9Hz),7.22(d,2H,J＝7.9Hz),4.51(d,2H,J＝5.9Hz),2.44(s,3H),2.37(s,3H)。
D)4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (oxiran-2-yl) biphenyl-3-carboxamide
A mixture of sodium hydride (60% in mineral oil, 0.85g,21.2mmol) in dimethyl sulfoxide (75mL) was cooled to-10 ℃. Trimethylsulfoxonium iodide (4.66g,21.2mmol) in DMSO (25mL) was added dropwise over a 10min period. The mixture was warmed to room temperature and stirred for an additional 1 hour. The mixture was cooled to 0 ℃ and 5-formyl-4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide (3.65g,10.6mmol) in DMSO (25mL) was added dropwise over a 10 minute period. The mixture was warmed to room temperature and stirred for 1 hour. The mixture was poured onto ice and extracted with ethyl acetate (3 × 150 mL). The combined extracts were dried over sodium sulfate and concentrated in vacuo. The mixture was purified by column chromatography using a dichloromethane to methanol gradient (2-10%) to give the title compound as a yellow oil. LC-MS 358.8[ M +1 ]]+。
E)5- (1-hydroxy-2-morpholinoethyl) -4' -methyl-N- ((6-methylpyridin-3-yl) methyl) biphenyl-3-carboxamide
In a 20mL reaction vessel 4' -methyl-N- ((6-methylpyridin-3-yl) methyl) -5- (oxiran-2-yl) biphenyl-3-carboxamide (15mg,0.042mmol), ethanol (2mL) and morpholine (13mg,0.15mmol) were combined. The mixture was heated at 50 ℃ overnight. Volatiles were removed under reduced pressure and the residue was purified by preparative HPLC (100X21.2mm C18 column, CH)3CN/Water [10mM Et2NH]) The title product is obtained.
LC-MS:446.2[M+1]+；1H NMR(400MHz,DMSO-d6):9.13(t,1H,J＝5.8Hz),8.40(d,1H,J＝2.3Hz),8.00(t,1H,J＝1.7Hz),7.80(s,1H),7.76(s,1H),7.62(d,3H,J＝8.2Hz),7.30(d,2H,J＝7.8Hz),7.26(d,1H,J＝7.8Hz),5.21(d,1H,J＝4.0Hz),4.87-4.80(m,1H),4.47(d,2H,J＝5.8Hz),3.56(t,4H,J＝4.6Hz),2.57-2.52(m,2H),2.48-2.44(m,4H),2.43(s,3H),2.35(s,3H)。
Representative compounds of the invention can be synthesized according to the methods set forth above and using appropriate reagents, starting materials, and purification methods well known to those skilled in the art.
Test of
Cell-based assays (e.g., calcium influx or electrophysiological assays), biochemical assays (e.g., with P2X) can be used2And P2X3Receptor binding assays) or compounds provided herein can be evaluated in models of pain or urinary function in animals. Examples of the tests are described below.
The purinergic receptor P2X is expressed in various tissues including various sensory and sympathetic ganglia, such as the Dorsal Root (DRG), the hypogonasal ganglion (ND), the trigeminal nerve (TG), and the supracervical ganglion (SCG), and also in smooth muscle cells2And P2X3(Burnstock, Trends Pharmacol. Sci.27:166-76, 2006). In several regions, P2X was co-expressed2And P2X3Receptor, functional studies confirmed the presence of heteropolymeric (heteromeric) P2X2/3Receptors whose properties differ from those of homomeric (homomeric) receptors. In addition, Inclusion and P2X are described3First transmembrane domain fused P2X2Chimeric P2X of N-terminal cytoplasmic domains2/3A receptor; these chimeric channels retain homomeric P2X3Pharmacological profile of the receptor with increased homomerisation of P2X2The non-desensitizing phenotype of the receptor (Neelands et al Br. J. Pharmacol.140:202-10, 2003). The non-desensitizing properties of the chimeric receptor are particularly useful in screening.
Members of the P2X family are ligand-gated, non-selective cation channels whose activity can be characterized by measuring calcium ion influx using electrophysiological methods or by using calcium-sensitive fluorescent dyes. The use of agonists such as ATP or ATP analogues such as α, β -methylene adenosine 5' -triphosphate (α β MeATP, Sigma-Aldrich) results in the opening of channels that generate current and calcium influx (Bianchi et al Eur.J.Pharmacol.376: 127-19, 1999).
Compounds provided herein can be tested for P2X by determining the ability of compounds provided herein to affect channel opening by ATP, α β MeATP or other agonists3And P2X2/3Antagonist activity of the receptor. Functional assays for receptor activity include, but are not limited to: (i) calcium ion influx as measured by fluorescence of calcium sensitive dyes; and (ii) ion influx due to channel opening as measured by electrophysiological methods. These methods can be used to assess channel function when the relevant receptor is heterologously expressed in mammalian or amphibian cells. These methods can also be used to evaluate the compounds provided herein in rodent primary neurons and other mammalian primary cells and cell lines that normally express a receptor of interest.
The compounds can also be further evaluated using biochemical means in binding to P2X3And P2X2/3Receptor side ability.
Compounds can also be evaluated for their ability to alter sensory and autonomic nervous system signaling in which receptors are known to have effects (e.g., bladder afferent signaling, sensory neuropathic pain). Finally, the compounds provided herein can be tested in vivo in relevant animal models known to those skilled in the art, such as a neuropathic, inflammatory or visceral pain model or a urinary incontinence model.
The following biological examples are provided to illustrate the compounds, pharmaceutical compositions and methods provided herein, but are not to be construed as limiting the scope thereof in any way.
Calcium uptake assay
Cloning and cell lines:
person P2X3(accession No. NM-002559), P2X2(accession No. NM-170682) and rat P2X3(accession No. NM-031075) and P2X2(accession No. NM-053656) into mammalian expression vectors (e.g.pcDNA5/TO or pcDNA3 Invitrogen). Human P2X was generated as described by Neelands et al2/3Chimeric clones were then cloned into expression vectors as described above. Receptors are expressed in cells such as HEK293 or 1321N1 (obtained from ECACC) by transient transfection using standard lipid-mediated transfection or by generating stable transfectants for each receptor. To express P2X2/3Heteromultimerizing receptor, P2X3Expression vectors were stably transfected into an already stably expressed P2X2The cell line of (1). Isolation of P2X Using pharmacological methods2/3Heteromeric function. The cell lines were maintained in DMEM + 5% Glutamax, appropriate levels of selective antibiotics and 10% heat-inactivated FBS.
P2X antagonist assay:
the functional activity of the compounds at the P2X receptor was determined by measuring the ability to inhibit agonist-induced calcium influx. Test Compound Pair P2X2/3Chimera, P2X3Homopolymers or P2X2/3Antagonist activity of heteromers. At the beginning of each screening day, agonist EC was determined50. Then using a predetermined agonist concentration (EC)50-90Cell line dependent) as a stimulus the% inhibition or IC50s of the compound was determined. The agonist used is α β MeATP, ATP or other ATP analogs. Compounds can be tested at concentrations of 1pM to 10. mu.M.
To test for antagonist activity, cells expressing the appropriate receptor were seeded in 96 or 384 well plates 18-24 hours prior to the assay. On the day of the experiment, 10mM CaCl was supplemented at most2Loading cells with calcium sensitive fluorescent dyes (e.g., Fluo-4no wash reagent-Invitrogen cat # F36206 or BD) in Hank buffered saline (HBSS)TMPBX Calcium Assay Kit-BD cat # 640175). Plates were incubated at 37 ℃ and then equilibrated at room temperature. Using a fluorescence imaging plate reader (e.g. FLIPR)TETRAMolecular Devices, Sunnyvale, CA) to determine agonist-induced calcium influx antagonism. The test comprises two stages: a pretreatment period followed by a treatment period. The compounds can be tested as follows. For the pretreatment period, 50 μ L of test compound in HBSS at a concentration of 3x was added to contain 100 μ L of dye loading mediumIn parenchymal cells, the compounds were tested to achieve a final concentration of 1 ×. For the treatment period, at set intervals (1-30 minutes) after pretreatment, 50 μ L of 1X test compound +4X agonist solution was added to yield final concentrations of 1X compound and 1X agonist. Fluorescence was measured at 0.1-3 second intervals-with an excitation wavelength of 494nM and an emission wavelength of 515 nM. The response was determined as (peak fluorescence after addition of the agonist) minus (baseline fluorescence before treatment). Percent inhibition was calculated as follows:
determination of IC by analysis of dose response data in 4-parameter Log-fitting Using GraphPad Prizm50The value is obtained.
Electrophysiological experiments
Whole-cell patch clamp:
whole cell recordings were performed using a multiclad 700A patch clamp amplifier and the Clampex acquisition program (Molecular Devices Corporation). Whole cell recordings were obtained with P2X3And/or P2X21321N1 or HEK cells stably or transiently transfected with the expression vector. The solution was applied for a 1-3 s period by a gravity flow 8-valve delivery system or for a millisecond period using a variable Dynaflow perfusion system (Cellectricon Inc.). The internal pipette solution may include 140mM cesium chloride, 10mM EGTA, and 5mM Hepes at ph 7.2; the standard external solution was 140mM NaCl, 5mM KCl, 1mM CaCl2、2mM MgCl225mM Hepes and 10mM glucose. Concentration-response curves were obtained by recording the current as a response to brief agonist application at 1-3min intervals, during which regular external solutions were perfused. To obtain an inhibition curve, the cells are pre-administered with antagonist for a defined period of time before the brief administration of agonist + antagonist. The duration of antagonist pre-application and agonist + agonist application was constant for the entire test concentration series. The agonist-induced current in the-60 or-80 millivolt-clamped cells was measured. By makingDetermination of IC Using GraphPad Prizm or Origin to analyze dose response data in a 4-parameter Log fit50The value is obtained.
Automatic two-electrode voltage clamp recording:
xenopus (Xenopus) oocytes (Nasco) were isolated by enzymatic isolation using collagenase (Worthington,2 mg/ml). The oocytes were then each injected with P2X3、P2X2Or P2X2And P2X3A combination of mRNAs. Each oocyte received 64nl of an aqueous RNA solution at a concentration of-0.01. mu.g/. mu.l. The injected oocytes were stored at 16 ℃ in a standard oocyte incubation solution ND96, containing (in mM) 96NaCl, 2KCl, 1MgCl2、1-5CaCl2And 50. mu.g/ml gentamicin. Agonist-induced currents resulting from the opening of the P2X channel were observed in oocytes 1-5 days after injection. For automatic recording, 8 oocytes were placed in the recording chamber. When filled with 3M KCl solution, each oocyte was penetrated with 2 glass electrodes having a resistance of 0.5-1 MOhm. Electrode advancement and oocyte penetration were under software control (OPUSXPRESS1.1, Molecular devices Corporation). Solutions were prepared in 96-well plates and robotically pipetted into the oocyte recording chambers using 8-channel pipettes. The inhibitory effect of the antagonist was determined by: the% residual current when the oocyte is stimulated with agonist in the presence of the test compound is calculated compared to the peak current in the presence of agonist alone. The order of applying the solutions to the oocytes was as follows: first a specific concentration (e.g. EC) is added50、EC80Or EC90) To elicit a maximal response. After pulsing, the oocytes were washed with ND96 for several minutes. A specific concentration of test compound is then added followed by the same concentration of compound and agonist. The concentration of the compound may be 0.3-10,000 nM. Determination of IC by analysis of dose response data in 4-parameter Log-fitting Using GraphPad Prizm or Origin software50The value is obtained.
Manual two-electrode voltage clamp:
each oocyte was manually punctured with 2 electrodes and agonist induced currents were measured using an oocyte clamp amplifier (Warner Instrument Corp.) and lemtex (molecular Devices corporation) acquisition software. The solution was delivered using gravity flow and applied as described above. Agonist-induced currents were measured in the absence and presence of antagonist. A series of concentrations of antagonist were tested to obtain an inhibition curve as described above.
Selective screening:
test inhibition P2X3And/or P2X2/3Activity of activated compounds at other P2X receptors to determine their selectivity for specific P2X family members. Examples of receptors tested include, but are not limited to, P2X1, P2X2P2X4, P2X5, P2X6 and P2X 7. The types of assays used for the selectivity determination may include: 1) agonist-induced calcium influx in cells heterologously expressing the relevant receptor; 2) electrophysiological measurement of receptor inhibition in mammalian cells heterologously expressing a receptor of interest or Xenopus oocytes. Method and data analysis and comparison of P2X above3And P2X2/3The same applies.
Radioligand binding:
radioligand experiments to determine the test compound vs P2X3Homomerisation and P2X2/3Affinity of heteromultimeric receptors. These studies also provide valuable insight into the mechanism of antagonism. For P2X3And P2X2/3General methods for radioligand binding assays for receptors are described by Jarvis et al J.Pharmacol. Exp.Ther.10:407-16, 2004.
Briefly, P2X was expressed transiently or stably3Or P2X2/3The cells of the recipient make cell membranes. Cells were grown to confluence, washed, separated, and stored as pellets at-80 ℃ until use. Some studies may require the addition of apyrase or hexokinase (Sigma-Aldrich) during cell membrane preparation to minimize ATP-mediated desensitization of receptors during membrane preparation. By resuspending the cell pellet in homogenization buffer, homogenizing, and centrifugingObtaining the membrane precipitate to prepare the membrane. The total protein concentration was determined using standard methods.
Alternative binding studies were performed using a revised approach from Jarvis et al. Under optimized conditions, in a binding buffer, radioligand ([ 3H)]A-317491, Abbott) or other high affinity radioligands and a range of different concentrations of test compounds. Ligand saturation studies were performed using a range of concentrations of radioligand. All binding reactions were terminated by rapid filtration through a glass fiber filter. The membranes were washed, incubated in scintillation fluid and counted in a scintillation counter. Determination of IC using 4-parameter logical Hill equation50The value is obtained.
Drug metabolism and pharmacokinetics
Caco-2 permeability:
caco-2 permeability was determined according to the method described in Yee, pharm. Res.14:763-6, 1997. Caco-2 cells were grown on a filter support (Falcon HTS porous addition system) for 14 days. The medium was removed from the top and bottom lateral chambers and the monolayers were preincubated with pre-warmed 0.3ml of top buffer and 1.0ml of bottom lateral buffer at 37 ℃ for 0.75 hours in a water bath shaker at 50 cycles/min. The top buffer was composed of Hanks balanced salt solution, 25mM D-glucose monohydrate, 20mM MES biological buffer, 1.25mM CaCl2And 0.5mM MgCl2(pH 6.5). The bottom outer buffer solution is composed of Hanks balanced salt solution, 25mM D-glucose monohydrate, 20mM HEPES biological buffer solution, 1.25mM CaCl2And 0.5mM MgCl2(pH 7.4). At the end of the preincubation, the medium was removed and a buffer solution (10 μ M) of the test compound was added to the top chamber. The addition was moved to wells containing fresh bottom outside buffer and incubated for 1 hr. The concentration of the drug in the buffer was determined by LC/MS analysis.
The flow rate (F, mass/time) is calculated from the slope of the occurrence of accumulation of the substrate on the receiver side, and the apparent permeability coefficient (Papp) is calculated according to the following equation:
Papp(cm/sec)＝(F*VD)/(SA*MD)
Wherein SA is the surface area of transport (0.3 cm)2) VD is the donor volume (0.3ml) and MD is the total amount of drug on the donor side when t ═ 0. All data represent the average of 2 additions. Monolayer integrity was determined from fluorescent yellow transport.
Human dofetilide binding:
cell paste of HEK-293 cells expressing HERG product can be suspended in 10-fold volume at 25 ℃ with a solution containing 1mM MgCl210mM KCl in 2M HCl was adjusted to pH7.5 in 50mM Tris buffer. Cells were homogenized using a Polytron homogenizer (20 seconds at maximum power) and centrifuged at 48,000g for 20 minutes at 4 ℃. The pellet was resuspended, homogenized and centrifuged again in the same manner. The resulting supernatant was discarded and the final pellet (10-fold volume of 50mM Tris buffer) was resuspended and homogenized at maximum power for 20 seconds. The membrane homogenate was aliquoted and stored at-80 ℃ until use. Aliquots were used for protein concentration determination using a protein assay rapid kit and an ARVO SX plate reader (Wallac). All operations, stock solutions and equipment were kept on ice at all times. For saturation experiments, experiments were performed with a total volume of 200. mu.l. By incubating 20 μ l of the [ sic ] in the absence or presence of a final concentration of 10 μ M of dofetilide (20 μ l) at room temperature3H]Doffilide and 160. mu.l of membrane homogenate (20-30. mu.g protein/well) for 60min to determine the saturation of total or non-specific binding, respectively. All incubations were terminated by Polyetherimide (PEI) -impregnated glass fiber filter paper, rapid vacuum filtration using a Skatron cell harvester, followed by two washes with 50mM Tris buffer (pH7.5 at 25 ℃). The radioactivity of receptor binding was quantified by liquid scintillation counting using a Packard LS counter.
For the competitive assay, compounds were diluted in a semi-log (semi-log) manner to 4-point dilutions on 96-well polypropylene plates. First, a total dilution with DMSO was performed, and then transferred to a container containing 1mM MgCl210mM KCl in 50mM Tris buffer (pH7.5 at 25 ℃) so that the final DMSO concentration is equal to 1%. Triplicate partitions on assay platesCompound (4. mu.l). Total binding and non-specific binding wells were set at 6 wells as vehicle and a final concentration of 10. mu.M dofetilide, respectively. Radioligand was prepared at a final concentration of 5.6x and the solution was added to each well (36 μ l). The assay was initiated by adding YSi poly-L-lysine Scintillation Proximity Assay (SPA) beads (50. mu.l, 1 mg/well) and membrane (110. mu.l, 20. mu.g/well). Incubation was continued at room temperature for 60 min. The plate was incubated at room temperature for a further 3 hours to allow the beads to settle. The receptor-bound radioactivity was quantified by counting the WALLAC microtob plate counter.
HERG test:
HEK293 cells stably expressing the HERG potassium channel were used for electrophysiological studies. Methods for stably transfecting such channels in HEK cells can be found elsewhere (Zhou et al Biophys. J.74:230-41, 1998). On the day before the experiment, cells were harvested from culture flasks and placed on glass coverslips in standard Minimal Essential Medium (MEM) containing 10% Fetal Calf Serum (FCS). The plated cells were stored at 37 ℃ and maintained at 95% O2/5％CO2In an incubator in an atmosphere. Cells were studied 15-28hr after collection.
HERG currents were studied in whole cell mode using standard patch clamp techniques. During this experiment, cells were superfused (superfused) with a standard external solution (mM) of the following composition: NaCl, 130; KCl, 4; CaCl2,2；MgCl21, 1; glucose, 10; HEPES, 5; pH7.4 with NaOH. Whole cell recordings were performed using a patch clamp amplifier and a patch pipette (patch pipette) with a resistance of 1-3MOhm when perfusing standard internal solutions of the following composition (mM): KCl, 130; MgATP, 5; MgCl21.0; HEPES, 10; EGTA5, pH7.2 and KOH. Accept only contact resistance (access resistance) and close resistance (seal resistance) with less than 15MOhm>Those cells of 1GOhm were used for further experiments. A series of resistance compensations (resistance compensation) were applied to 80% maximum. No leakage subtraction was performed. However, acceptable contact resistance depends on the recorded current magnitude and the series of resistance compensation levels that can be safely used. After the whole cell construction is completed and the cells are performed using pipette solution Sufficient time for dialysis: (>5min), a standard voltage protocol was applied to the cells to induce membrane currents. The voltage scheme is as follows. The membrane was depolarized from a holding potential of-80 mV to +40mV1000 ms. This step is followed by a voltage ramp (voltage ramp) down (rate 0.5mV msec-1) back to the holding potential. This voltage protocol was applied to the cells every 4 seconds continuously throughout the experiment (0.25 Hz). The peak current amplitude of about-40 mV induced during the ramp (ramp) was measured. Once a stably elicited current response is obtained in the external solution, the vehicle (standard external solution of 0.5% DMSO) will be applied for 10-20min with a peristaltic pump. Test compounds were applied for a period of 10min at 0.3, 1, 3 or 10mM as long as the resulting change in current response amplitude was minimal under the vehicle control conditions. The 10min period included providing time for the solution to pass from the solution reservoir through the tubing to the recording chamber via the pump. After the concentration of the drug in the chamber has sufficiently reached the test concentration, the cells are contacted with the compound solution for more than 5 min. Followed by a wash period of 10-20min to assess reversibility. Cells were finally exposed to high doses of dofetilide (5mM), a specific IKr blocker, to assess insensitive endogenous currents.
All experiments were performed at room temperature (23. + -. 1 ℃). The resulting membrane currents were recorded by computer on-line operation, filtered at 500-1KHz (Bessel-3dB) using a chip-clamp amplifier and special data analysis software, and sampled at 1-2 KHz. The peak current amplitude occurring at about-40 mV was determined off-line using a computer.
The arithmetic mean of the 10 amplitude values was calculated under vehicle control conditions and in the presence of the drug. The percentage of IN reduction IN each experiment was obtained from the calibrated current values using the following formula: IN ═ 1-ID/IC) x100, where ID is the average of the current IN the presence of drug and IC is the average of the current under control conditions. Separate experiments were performed for each drug concentration or time paired control, and the arithmetic mean of each experiment was defined as the study result.
Half-life in Human Liver Microsomes (HLM):
test compound (1. mu.M) was mixed with 3.3mM MgCl2And 0.78mg/mL of HLM (HL101) were incubated together in 100mM potassium phosphate buffer (pH7.4) at 37 ℃ in a 96-deep well plate. The reaction mixture was divided into two groups: non-P450 and P450 groups. NADPH was added only to the reaction mixtures of the P450 group. Aliquots of the P450 set of samples were taken at time points 0, 10, 30 and 60min, where the 0min time point represents the time when NADPH was added to the P450 set of reaction mixtures. Aliquots of the non-P450 group samples were taken at-10 and 65min time points. The collected aliquots were extracted using acetonitrile solution containing an internal standard. The precipitated protein was spun down with a centrifuge (2000rpm,15 min). The concentration of the compound in the supernatant was determined by an LC/MS/MS system. Half-life values were obtained by plotting the natural logarithm of the peak area ratio of compound/internal standard versus time. The metabolic rate (k) is generated by the slope of the best fit line for each point. This was converted to a half-life value using the following equation:
Half-life is ln 2/k.
In vivo efficacy assay
P2X can be tested in various animal models of human disease3、P2X2/3Antagonists, including models of neuropathic, inflammatory and visceral pain and models of bladder function. P2X administration before or after induction model depending on the specific model and PK profile of compound3An antagonist. Routes of administration may include intraperitoneal (i.p.), subcutaneous (s.c.), oral (p.o.), intravenous (i.v.), intrathecal (i.t.), or intraplantar. Endpoints for these studies may include mechanical allodynia, thermal hyperalgesia, cold allodynia, formalin-induced reduction in pain response, reduced or altered mechanical sensation of the bladder with writhing and contractions, appropriate for the model described below.
Formalin model:
test compounds were administered at different times prior to intraplantar administration of formalin. A dilute solution of formalin (25-50 μ L of 1-2.5% formaldehyde/saline) was administered s.c. into the plantar surface of the left hind paw under mild compression. Immediately after injection, the animals were placed on a sieve table in a clean observation room, large enough to allow the animals to move freely during the study. Behavior is scored using manual scoring or automatic scoring.
And (3) manual scoring: using a three-channel timer, the observer notes the drop in support weight (t)1) Lifting the claw (t)2) And licking/biting/shaking (t)3) Time (t in seconds). According to the method of Dubuisson and Dennis, Pain,4:161-174,1977, applying the formula t1+2t2+3t3The results are weighted by/180, where 180s is the estimated time for each increment. The acquisition behavior is incremented by 3min alternately, with a start time of 0min (i.e., 0-3min,6-9min, etc.), and ending at 60 min.
Automatic scoring: a small metal strip weighing 0.5g was placed on the left paw. Formalin was given and the animals were placed, without restriction, inside a viewing chamber using an electromagnetic detector system (Automated probability Analyzer, University of California, San Diego). The number of paw withdrawals was recorded electronically.
ATP and α β -methylene ATP (α β meATP) -induced inflammatory pain:
rats were dosed subcutaneously with up to 1 μ Mol α β meATP, ATP, adenosine or PBS in volumes up to 100 μ L into the dorsal aspect of the hind paw. Immediately after injection, the animals were placed inside a clear observation chamber, which was large enough to allow free movement of the animals. The duration of withdrawal and licking was recorded at 20 intervals to evaluate nocifensive behavior. Responses were determined using either manual or automated methods as described above for the formalin test. Other behavioral tests may include the assessment of mechanical allodynia and thermal hyperalgesia. For testing, compounds were administered prior to agonist injection.
Complete adjuvant model freund (CFA):
animals received 100 μ L of complete freund's adjuvant s.c. injection under isoflurane anesthesia into the plantar surface of the right hind paw containing 100 μ g of Mycobacterium tuberculosis (Mycobacterium tuberculosis) strain H37 Ra. Swelling and inflammation were visible within 1h after administration. Nociceptive testing can begin 24h after CFA administration. The compound is typically administered 0.5-12hr prior to testing.
Carrageenin-induced acute pain:
animals received 100 μ L of 2% carageenan under isoflurane anesthesia and were injected subcutaneously into the plantar surface of the right hind paw. Swelling and inflammation were visible within 1h after administration. Nociceptive testing can begin 3-24h after carrageenan administration (Hargreaves et al Pain,32:77-88,1988). The compound is typically administered 0.5-12hr prior to testing.
Chronic constrictive injury model (CCI or Bennett model):
the CCI model was completed according to the methods described by Bennett and Xie, Pain,33:87-107,1988. Briefly, under isoflurane anesthesia, the right sciatic nerve was exposed horizontally in the mid-thigh by blunt dissection through the biceps femoris muscle. The proximal part of the sciatic nerve bifurcation, i.e. the nerve of about 7mm, has no adherent tissue, and 4 loose ligated 4.0 chromium gut sutures are tied up around the nerve. The space between ligations was about 1 mm. The wound is closed in layers and the skin is closed with staples or non-silk sutures. Animals simulating surgery were treated in the same manner except that the sciatic nerve was not ligated. Nociceptive testing may be performed 7-21 days after surgery. The compound is typically administered 0.5-12hr prior to testing.
Spinal nerve transection (SNT or Chung model):
under anesthesia, rats were placed in a prone position on a sterile plane. A median incision of L4-S2 was performed and the left paraspinal muscles were isolated from the spinous process. The L5 and L6 spinal nerves were firmly ligated with 4-0 silicon-treated silk suture according to the method described by Kim and Chung, Pain,50:355-363, 1992. The L4 spinal nerve was carefully protected to prevent surgical injury. The skin was closed with wound clips and the animals were returned to their cages. Rats that showed postoperative neurological deficits or difficulty in grooming the prolongation were excluded from the experiment. Animals were evaluated for nociceptive response (baseline) prior to surgery and then at various time points after administration of the test compound. Nociceptive testing may be performed 7-21 days after surgery. The compound is typically administered 0.5-12hr prior to testing.
Chemotherapy-induced painful neuropathy:
chemotherapy neuropathy was induced by i.p. administration of 1mg/kg Taxol (Taxol) administered once a day on an alternating 4-day basis (Polomano et al Pain,94:293-304, 2001). Nociceptive testing may be performed 9-30 days after the start of taxol administration. The compound is typically administered 0.5-12hr prior to testing.
And (3) nociceptive testing:
Mechanical allodynia: mechanical allodynia testing was performed using the up-down method (up-down method) of Dixon, Ann.Rev.Pharmacol.Toxicol.20:441-462,1980, modified to the mechanical threshold of Chaplan et al J.Neurosci.Methods53:55-63,1994. To evaluate tactile allodynia, rats were placed in clear plexiglas chambers with a mesh floor and allowed to acclimatize for a period of at least 15 minutes. Following acclimation, a series of von Frey monofilaments were provided to the plantar surface of the left (manipulated) foot of each rat. Each presentation was continued for a period of 4-8 seconds or until nociceptive flinching behavior was observed. Flinching, paw flinching or paw licking were considered nociceptive behavior responses. The 50% withdrawal threshold was calculated using the method described by Chaplan et al J.Neurosci.Methodss 53:55-63,1994.
Thermal hyperalgesia: hindpaw withdrawal latencies to noxious thermal stimuli were determined using a plantar tester (Ugo Basile) according to the technique described by Hargreaves et al, Pain32:77-88,1988. Radiant heat was concentrated on the plantar surface of the ipsilateral paw, and paw withdrawal latency was measured. An increase in paw withdrawal latency indicates reversal of hyperalgesia.
Mechanical pain and hypersensitivity: the paw pressure test can be used to assess mechanical hyperalgesia. For the purpose of this test, hindpaw withdrawal thresholds (PWT) against noxious mechanical stimuli were determined using an analgesymeter (Ugo Basile) as described by Stein et al Pharmacol, biochem, Behav.31:451-455, 1988. The maximum weight that can be applied to the hind paw was set at 250g, and the endpoint was defined as the complete withdrawal of the paw. PWT was measured once at each time point for each rat.
Cold allodynia: to determine cold allodynia, 1 drop of acetone was applied to the plantar surface of the paw using a 50 μ L hamilton syringe through the underside of the grid on which the animal stood. This process was performed 5 times with 3min intervals. Vigorous shaking was recorded as a positive response and the time taken for shaking was recorded. Alternatively, cold allodynia can be tested using a cold water bath method in which the animals are placed in a cold water bath at a depth of 1.5-2.0cm and a temperature of 3-4 degrees celsius and the number of times the paw is lifted is counted.
Colorectal distension (CRD):
animals were fasted prior to induction model, but water was taken ad libitum for 16h before induction model. The 5cm latex balloon was connected to a barostat system consisting of a flow meter and pressure control program through the length of the tubing. Under isoflurane anesthesia, the balloon was inserted transanally into the distal colon a distance of 5cm from the anus and taped to the base of the tail. After anaesthesia, animals were placed ad libitum in clean polypropylene cages and allowed to acclimate for 30 min. The balloon was inflated stepwise from 0-75mmHg every 30s in 5mm increments. The colonic response threshold is defined as the pressure that induces the first abdominal contraction. Abdominal contractions, indicative of visceral pain, are associated with a hump, a humped position, licking the lower abdomen, repeated contraction waves of the ipsilateral obliterator system with inward rolling, stretching, pressing the lower abdomen to the floor along with the lateral hind limbs (Wesselman, neurosci. lett.,246:73-76,1998). Alternatively, electrodes may be placed in the extraabdominal oblique muscle system for electromyographic recording of abdominal contractions. In this regard, EMG activity is quantified during colonic balloon inflation. The compound is typically administered 0.5-12hr prior to testing.
Acetic acid writhing test:
rats were given i.p. 0.6% acetic acid solution (10ml/kg) and the number of abdominal contractions within 30min was counted. The compound is typically administered 0.5-12hr prior to testing.
Bladder afferent recording:
to determine P2X3And P2X2/3Receptor inhibition has a precise role in the micturition response, testing compounds to modulate afferent signaling from the bladder. Compounds were evaluated in bladder/pelvic nerve preparations as described in Vlaskovska et al J.neuroscience,21: 5670-. Briefly, the periurethral canal connecting to the inferior vertebra and surrounding tissue was isolated in its entirety and oxygenated with oxygen (5% CO) in the recording chamber2And 95% O2) It was superfused with Krebs solution. The bladder is catheterized through the urethra for intraluminal infusion. A second double lumen catheter was inserted into the bladder to measure intraluminal pressure and empty the bladder. After the bladder is prepared, the pelvic nerve, which is removed from the vertebrae, is dissected away and a suction glass electrode is inserted. Neural activity was measured using standard electrophysiological methods. After a stabilization period of 60min, iterative ramp expansion (ramp expansion) was performed until the afferent response stabilized. This stable afferent response was used to compare the mechanical sensitivity of bladder afferents between different treatment groups.
Isochoric bladder contraction test:
female sprague-dawley rats were anesthetized, the trachea was dissected, and cannulae were inserted in the carotid artery and femoral vein. The bladder is accessed by laparotomy and the ureters are ligated and transected. For fluid infusion and pressure measurement, the bladder is cannulated.
After surgery, the bladder is infused with saline until a stable volume-induced bladder contraction is induced. Once a stable threshold volume and contraction frequency are obtained, the compound is administered to the animal and the contraction frequency is determined.
Bladder function reperfusion and cystitis models:
animals were anesthetized and transurethral closed bladder manometry was performed as described above (Dmitrieva et al Neuroscience78:449-59, 1997; Cockayne et al Nature407:1011-5, 2000). A PE-10 polypropylene catheter was inserted into the bladder via the urethra. Each cystometrogram consists of a slow infusion of saline into the bladder through a transurethral catheter, followed by recording of the pressure associated with the infusion by a pressure transducer. Contractions greater than a predetermined threshold are interpreted as micturition contractions. For each cystometrogram, the volume at which active contraction occurred (micturition threshold) and the number of contractions per cystometrogram were recorded. The effect of the compound was then determined.
Cystometrograms can also be obtained in animal models of cystitis in which the bladder is stimulated by injection of cyclophosphamide (150mg/kg, i.p.) 24hr prior to cystometry or by infusion of up to 1% acetic acid during cystometry.
The synthetic and biological examples described in this application are provided to illustrate the compounds, pharmaceutical compositions and methods provided herein, but are not intended to limit the scope thereof in any way. In these examples, all temperatures are in degrees Celsius (unless otherwise indicated). Compounds that can be prepared according to the methods provided herein are shown in the following table along with their biological activity data. Representative compounds were synthesized according to the methods described above.
Exemplary Compounds provided herein
The following compounds were or may be prepared according to the synthetic methods described herein. Calcium uptake assays were performed as described above and the activity of each compound was expressed as follows:
+ Compounds hP2X2/3H IC50>1000nM
The + Compound shows hP2X2/3H IC50501-1000nM
The + + + Compound shows hP2X2/3H IC50100-500nM
The + + + + + Compound shows hP2X2/3H IC50<100
Compounds display hP2X3IC50>1000nM
Compounds display hP2X3IC50501-1000nM
Compounds display hP2X3IC50100-500nM
Compounds display hP2X3IC50<100
TABLE 1 Ca influx IC of typical Compounds50
From the foregoing description, it will be apparent to those skilled in the art that various modifications and variations can be made in the compositions and methods provided herein. All such modifications as fall within the scope of the appended claims are intended to be included therein.
All publications (including, but not limited to, patents and patent applications) cited in this specification are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as though fully set forth.
At least some of the chemical names of the compounds of the present invention specified and recited in this application were generated on an automated basis by using a commercially available chemical naming software program and were not independently validated. Representative programs that perform this function include the Lexichem naming tool sold by Open Eye Software, inc and the Autonom Software tool sold by MDL, inc. In the case where the chemical names shown and the structures described differ, the structures shown control.
Use ofthe/DRAW is made into the chemical structure shown herein. Any open valency appearing on a carbon, oxygen or nitrogen atom in a structure herein indicates the presence of a hydrogen atom, wherein a chiral center is present in the structure, but does not show specific stereochemistry for the chiral center, and enantiomers associated with chiral structures are encompassed by the structure.