Disclosure of Invention
The present invention aims to solve, at least to some extent, one of the above technical problems or at least to provide a useful commercial choice.
According to one aspect of the present invention, there is provided a compound which is a compound of formula (I), or a tautomer, stereoisomer, hydrate, solvate, salt or prodrug of a compound of formula (I):
Wherein:
R1、R2、R3 or R4 are independently selected from the group consisting of-H, -CN, halogen, C1-6 alkyl, C3-6 cycloalkyl optionally substituted with one or more Ra, Ra is selected from the group consisting of halogen, C1-6 alkyl;
Each R5 is independently selected from H, -CN, halogen, C1-6 alkyl, C1-6 alkoxy, C3-6 cycloalkyl, optionally substituted with one or more Rb, Rb is selected from halogen, C1-6 alkyl;
At least 1 of M1、M2、M3、M4、M5 is-C (R6) =or-ch=, the remainder are-N (R7) -or-n=, and at least 1 is-N (R7) -or-n=;
R6 and R7 are the same or different and are independently selected from the group consisting of C1-6 alkyl, C1-6 alkoxy, C3-6 cycloalkyl, C3-6 cycloalkoxy, absent, -H, -CN, halogen, unsubstituted or optionally substituted with one or more Rc, said Rc being selected from halogen, C1-6 alkyl, preferably said Rc is selected from-F, -Cl, methyl, ethyl, propyl;
Z is selected from the group consisting of-O-, -S-, and,
L is a single bond or is selected from the group consisting of C1-10 straight chain alkylene, C3-10 cycloalkylene, 3-10 membered heterocyclylene, unsubstituted or optionally substituted with one or more Rd, Rd is selected from halogen, methyl, ethyl, cyclopropyl;
Q is C3-C10 cycloalkyl, 3-10 membered heterocyclyl, C6-C10 aryl, 5-10 membered heteroaryl, optionally substituted with one or more R5;
m is selected from the integers of 0-6, n is selected from the integers of 0-6, and p is selected from the integers of 0-5.
In some embodiments of the invention, Ra is selected from the group consisting of-F, -Cl, methyl, ethyl, propyl, and the remaining variables are as defined herein.
In some embodiments of the invention, Rb is selected from the group consisting of-F, -Cl, methyl, ethyl, propyl, and the remaining variables are as defined herein.
In some embodiments of the invention, R6 and R7 are independently selected from the group consisting of absent, -H, -CH3、-CH2CH3、-CH2CH2CH3,-F, -Cl, -Br, -CF3, the remaining variables being as defined herein.
In some embodiments of the invention, R6 and R7 are independently selected from the absence, -H, -CH3、-F、-Cl、-CF3, and the remaining variables are as defined herein.
In some embodiments of the invention, L is methylene, ethylene, cyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, azetidinylene, methylene-cyclopropyl, methylene-cyclobutylene, methylene-cyclopentylene, methylene-cyclohexylene, methylene-azetidinylene, the remaining variables being as defined herein, optionally substituted with one or more Rd.
In some embodiments of the invention, Q is selected from cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, azetidinyl, pyrrolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, morpholinyl, phenyl, indenyl, naphthyl, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, furanyl, thienyl, thiazolyl, oxazolyl, pyridinyl, pyrimidinyl, pyrazinyl, pyridazinyl, benzimidazolyl, indolyl, or quinolinyl, the remaining variables being as defined herein.
In some embodiments of the invention, Q is selected from benzene rings and the remaining variables are as defined herein.
In some embodiments of the invention, at least 2 or 3 of M1、M2、M3、M4、M5 are-C (R6) = or-ch=, the remainder are-N (R7) -or-n=, and at least 1 are-N (R7) -or-n=, wherein R6 and R7 are as defined above and the remaining variables are as defined herein.
In some aspects of the present invention,Selected from the group consisting ofOr (b)Wherein M3 and M5 are independently selected from-C (R6) =, -ch=or-nh=, wherein R6 and R7 are as defined above and the remaining variables are as defined herein.
In some aspects of the present invention,Selected from the group consisting ofOr (b)Or (b)Or (b)Or (b)Or (b)The remaining variables are as defined herein.
In some embodiments of the invention, in formula (I), m=0, 1 or 2;n =0, 1 or 2, the remaining variables are as defined herein.
In some embodiments of the invention, R1、R2、R3 or R4 in formula (I) is independently selected from the group consisting of-H, -CN, methyl, ethyl, n-propyl, isopropyl, optionally substituted with 1-3C1-3 alkyl groups Ra, Ra is selected from the group consisting of-F, -Cl, C1-3 alkyl, and the remaining variables are as defined herein. In some embodiments of the invention, in formula (I), each R1、R2、R3 or R4 is independently selected from-H, -CH3、-CH2CH3、-CH2CH2CH3, and the remaining variables are as defined herein.
In some aspects of the invention, in formula (I),Selected from the group consisting ofThe remaining variables are as defined herein.
In some embodiments of the invention, in formula (I), L is selected from: the remaining variables are as defined herein.
In some embodiments of the invention, in formula (I), p=0, 1 or 2, the remaining variables are as defined herein.
In some embodiments of the invention, each R5 is independently selected from H、-CN、-F、-Cl、-Br、-CH3、-CF3、-OCHF2、-OCF3、-CH2CH3、-CH2CH2CH3、The remaining variables are as defined herein.
In some aspects of the present invention, each R5 is independently selected from the group consisting of-H, -CN-F, -Cl, -Br, -CF3、-OCF3、-OCHF2, the remaining variables are as defined herein.
In some aspects of the invention, in formula (I),Selected from the group consisting ofThe remaining variables are as defined herein.
According to an embodiment of the present invention, the compounds of the present invention comprise a compound of the formula, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of a compound of the formula:
According to an embodiment of the present invention, the compounds of the present invention comprise a compound of the formula, or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of a compound of the formula:
the compounds of the present invention may exhibit tautomerism. The present invention includes all tautomeric forms of a compound, whether in equilibrium or one form predominates, and the invention encompasses each tautomeric form.
According to a further aspect of the present invention, there is provided a pharmaceutical composition comprising a therapeutically effective dose of at least one compound of formula (I) according to the present invention or a pharmaceutically acceptable salt, tautomer, stereoisomer, hydrate, solvate or prodrug thereof.
"Pharmaceutical composition" means a mixture of one or more of the compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof with other chemical components, such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.
According to a further aspect of the present invention, there is provided the use of a compound of formula (I), a pharmaceutically acceptable salt, tautomer, stereoisomer, hydrate, solvate, or prodrug thereof, or a pharmaceutical composition comprising a compound of formula (I), or a pharmaceutically acceptable salt, tautomer, stereoisomer, hydrate, solvate, or prodrug thereof, in the manufacture of a medicament for the treatment of a disease associated with ATX.
In some embodiments of the invention, wherein the ATX-related disease is selected from cancer, metabolic disease, renal disease, liver disease, fibrotic disease, interstitial lung disease, proliferative disease, inflammatory disease, pain, autoimmune disease, respiratory disease, cardiovascular disease, neurodegenerative disease, dermatological disorder, and/or abnormal angiogenesis-related disease.
In some embodiments of the invention, wherein the ATX-related disease is selected from interstitial lung disease, pulmonary fibrosis, liver fibrosis, kidney fibrosis, preferably, from idiopathic pulmonary fibrosis. According to embodiments of the present invention, the compounds of the present invention are of significant advantage in the treatment of pulmonary fibrosis, particularly idiopathic pulmonary fibrosis.
In some embodiments of the invention, wherein the ATX-related disease is selected from metabolic diseases.
In some embodiments of the invention, the metabolic disorder is selected from the group consisting of type II diabetes, non-alcoholic steatohepatitis. According to embodiments of the present invention, the compounds of the present invention are of significant advantage in the treatment of metabolic disorders, particularly type II diabetes mellitus, non-alcoholic steatohepatitis.
In some embodiments of the invention, wherein the ATX-related disorder is selected from neuropathic pain, inflammatory pain, preferably from osteoarthritis-related pain. According to embodiments of the present invention, the compounds of the present invention have significant advantages in the treatment of pain associated with osteoarthritis.
In some embodiments of the invention, wherein the ATX-related disease is selected from cancer. According to embodiments of the present invention, the compounds of the present invention have significant advantages in the treatment of cancer.
Definition and description of terms
Unless otherwise indicated, the radical and term definitions recited in the specification and claims of the present application, including as examples, exemplary definitions, preferred definitions, definitions recited in tables, definitions of specific compounds in the examples, and the like, may be arbitrarily combined and coupled with each other. Such combinations and combinations of radical definitions and structures of compounds should fall within the scope of the present description.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the claimed subject matter belongs. All patents, patent applications, and publications cited herein are hereby incorporated by reference in their entirety unless otherwise indicated. If there are multiple definitions of terms herein, the definitions of this chapter shall control.
Conventional methods within the skill of the art, such as mass spectrometry, NMR, IR and UV/Vis spectrometry and pharmacological methods, are employed unless otherwise indicated. Unless specifically defined otherwise, the terms used herein in the description of analytical chemistry, organic synthetic chemistry, and pharmaceutical chemistry are known in the art. Standard techniques may be used in chemical synthesis, chemical analysis, pharmaceutical preparation, formulation and delivery, and treatment of patients. For example, the reaction and purification can be carried out using the manufacturer's instructions for the kit, or in a manner well known in the art or in accordance with the teachings of the present application. The techniques and methods described above may generally be practiced according to conventional methods well known in the art, based on a number of general and more specific descriptions in the literature cited and discussed in this specification. In this specification, groups and substituents thereof can be selected by one skilled in the art to provide stable moieties and compounds. When substituents are described by conventional formulas written from left to right, the substituents also include chemically equivalent substituents obtained when writing formulas from right to left. For example, CH2 O is equivalent to OCH2.
Where a range of values recited in the specification and claims is understood to be an "integer," it is understood that both ends of the range and each integer within the range are recited. For example, an "integer of 1 to 6" should be understood to describe each integer of 0, 1, 2, 3, 4, 5, and 6. When a numerical range is understood as a "number," it is understood that both endpoints of the range are noted, as well as each integer within the range, and each fraction within the range. For example, a "number of 1 to 10" should be understood to describe not only each integer of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10, but also at least the sum of each integer with 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, respectively.
The term "pharmaceutically acceptable" is intended to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio.
The term "pharmaceutically acceptable salt" refers to pharmaceutically acceptable salts of non-toxic acids or bases, including salts of inorganic acids and bases, organic acids and bases.
In addition to pharmaceutically acceptable salts, other salts are contemplated by the present invention. They may serve as intermediates in the purification of the compounds or in the preparation of other pharmaceutically acceptable salts or may be used in the identification, characterization or purification of the compounds of the invention.
The term "stereoisomer" refers to an isomer produced by the spatial arrangement of atoms in a molecule, and includes cis-trans isomers, enantiomers, non-corresponding isomers and conformational isomers. The stereochemical definitions and conventions used in the present invention are generally defined as S.P.Parker,Ed.,McGraw-Hill Dictionary of Chemical Terms(1984)McGraw-Hill Book Company,New York;and Eliel,E.and Wilen,S.,"Stereochemistry of Organic Compounds",John Wiley&Sons,Inc.,New York,1994.
Depending on the choice of starting materials and methods, the compounds according to the invention may be present in the form of one of the possible isomers or mixtures thereof, for example as pure optical isomers or as isomer mixtures, for example as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. When describing optically active compounds, the prefix D and L or R and S are used to denote the absolute configuration of the molecule in terms of chiral center (or chiral centers) in the molecule. The prefixes D and L or (+) and (-) are symbols for designating the rotation of plane polarized light by a compound, where (-) or L represents that the compound is left-handed. The compound prefixed with (+) or D is dextrorotatory. For a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be referred to as enantiomers, and mixtures of such isomers are generally referred to as mixtures of enantiomers. The 50:50 mixture of enantiomers is referred to as a racemic mixture or racemate, which may occur when there is no stereoselectivity or stereospecificity in the chemical reaction or process. Many geometric isomers of olefins, c=n double bonds, etc. may also be present in the compounds described herein, and all such stable isomers are contemplated in the present invention. When the compounds described herein contain olefinic double bonds, such double bonds include E and Z geometric isomers unless specified otherwise. If the compound contains a disubstituted cycloalkyl, the cycloalkyl substituent may be in cis or trans (cis-or trans-) configuration.
When the bonds to chiral carbons in the formulae of the present invention are depicted in straight lines, it is understood that both the (R) and (S) configurations of the chiral carbons and the enantiomerically pure compounds and mixtures thereof resulting therefrom are included within the general formula. The graphic representation of racemates or enantiomerically pure compounds herein is from Maehr, J.chem. Ed.1985,62:114-120. Unless otherwise indicated, the absolute configuration of a stereocenter is indicated by the wedge-shaped key and the dashed key.
Optically active (R) -or (S) -isomers can be prepared using chiral synthons or chiral preparations, or resolved using conventional techniques. The compounds of the invention containing asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Resolution of the racemic mixture of the compounds may be carried out by any of a number of methods known in the art. An exemplary method includes fractional recrystallization using a chiral resolving acid that is an optically active salified organic acid. Suitable resolving agents for use in the fractional recrystallisation process are, for example, D and L forms of optically active acids such as tartaric acid, diacetyl tartaric acid, dibenzoyl tartaric acid, mandelic acid, malic acid, lactic acid or various optically active camphorsulphonic acids such as β -camphorsulphonic acid. Other resolving agents suitable for the fractional crystallization process include stereoisomerically pure forms of α -methyl-benzylamine (e.g., S and R forms or diastereoisomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methyl ephedrine, cyclohexylethylamine, 1, 2-diaminocyclohexane, and the like. Resolution of the racemic mixture may also be carried out by eluting on a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). High Performance Liquid Chromatography (HPLC) or Supercritical Fluid Chromatography (SFC) may be used. The choice of the particular method and elution conditions, choice of chromatographic column can be selected by one skilled in the art based on the structure of the compound and the results of the assay. Further, any enantiomer or diastereomer of a compound described herein may also be obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration.
The term "tautomer" refers to a functional group isomer that results from the rapid movement of an atom in a molecule at two positions. The compounds of the present invention may exhibit tautomerism. Tautomeric compounds may exist in two or more interconvertible species. Proton-mobile tautomers result from the migration of a hydrogen atom covalently bonded between two atoms. Tautomers generally exist in equilibrium and attempts to isolate individual tautomers often result in a mixture whose physicochemical properties are consistent with the mixture of compounds. The location of the equilibrium depends on the chemical nature of the molecule. For example, among many aliphatic aldehydes and ketones such as acetaldehyde, the keto form predominates, while among phenols, the enol form predominates. The present invention encompasses all tautomeric forms of the compounds.
In examples of the invention, protons may occupy cyclic forms of two or more positions of the heterocyclic system, for example, 1H-and 3H-imidazoles, 1H-, 2H-and 4H-1,2, 4-triazoles, 1H-and 2H-isoindoles, and 1H-and 2H-pyrazoles. Tautomeric forms may be in equilibrium or spatially fixed to one form by appropriate substitution. For example:
the nomenclature differs because the hydrogen of the nitrogen on the triazole can be on any of the three nitrogen due to resonance, but these three forms represent one compound in fact.
The term "pharmaceutical composition" means a mixture of one or more of the compounds described herein or a physiologically/pharmaceutically acceptable salt or prodrug thereof with other chemical components, such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to the organism.
For a drug or pharmacologically active agent, the terms "effective dose", "effective amount" or "therapeutically effective amount" refer to a sufficient amount of the drug or agent that is non-toxic but achieves the desired effect. For the purposes of the present oral dosage form, an "effective amount" of one active agent in a composition refers to that amount which is required to achieve the desired effect when used in combination with another active agent in the composition. Determination of an effective amount varies from person to person, depending on the age and general condition of the recipient, and also on the particular active substance, a suitable effective amount in an individual case can be determined by one skilled in the art according to routine experimentation.
The term "active ingredient", "therapeutic agent", "active substance" or "active agent" refers to a chemical entity that is effective in treating a disorder, disease or condition of interest.
The term "solvate" refers to a compound of the invention or a salt thereof that includes a stoichiometric or non-stoichiometric solvent that binds with non-covalent intermolecular forces, and when the solvent is water, is a hydrate.
The term "prodrug" refers to a compound of the invention that can be converted to a biologically active compound under physiological conditions or by solvolysis. Prodrugs of the invention are prepared by modifying functional groups in the compounds, which modifications may be removed by conventional procedures or in vivo to give the parent compound. Prodrugs include compounds wherein a hydroxyl group or amino group of a compound of the invention is attached to any group that, when administered to a mammalian subject, cleaves to form a free hydroxyl group, free amino group, respectively.
The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more of the atoms comprising the compounds. For example, compounds may be labeled with a radioisotope, such as deuterium (2 H), tritium (3 H), iodine-125 (125 I) or C-14 (14 C). All isotopic variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.
The term "C1-6 alkyl" is understood to mean preferably a straight or branched saturated monovalent hydrocarbon radical having 1 to 6 carbon atoms. The alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-ethylpropyl, 1, 2-dimethylpropyl, neopentyl, 1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3-dimethylbutyl, 2-dimethylbutyl, 1-dimethylbutyl, 2, 3-dimethylbutyl, 1, 3-dimethylbutyl, or 1, 2-dimethylbutyl, or the like, or an isomer thereof. In particular, the groups have 1,2 or 3 carbon atoms ("C1-C3 alkyl"), such as methyl, ethyl, n-propyl or isopropyl.
The term "C3-6 cycloalkyl" is understood to mean a saturated monovalent monocyclic or bicyclic hydrocarbon ring having 3 to 6 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.
The term "alkoxy" refers to an alkyl group as defined above having the indicated number of carbon atoms attached through an oxygen bridge. Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and sec-pentoxy.
The term "C1-6 alkoxy" is understood to mean-O- (C1-6 alkyl), where "C1-6 alkyl" has the above definition.
The term "C3-6 cycloalkoxy" is understood to be-O- (C3-6 cycloalkyl) in which "C1-6 alkyl" has the above definition.
The term "halo" or "halogen" as used herein refers to fluorine, chlorine, bromine and iodine.
"Haloalkyl" is meant to include branched and straight-chain saturated aliphatic hydrocarbon groups having the specified number of carbon atoms, substituted with one or more halogens (e.g., -CvFw, where v=1 to 3,w =1 to (2v+1)). Examples of haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl, pentachloroethyl, 2-trifluoroethyl, heptafluoropropyl, and heptachloropropyl.
The term "3-10 membered heterocyclyl" means a saturated monovalent monocyclic or bicyclic hydrocarbon ring comprising 1 to 5, preferably 1 to 3 heteroatoms selected from N, O and S. In particular, the heterocyclic groups may include, but are not limited to, 4-membered rings such as azetidinyl, oxetanyl, 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl, or 6-membered rings such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl, or trithianyl, or 7-membered rings such as diazepinyl. Optionally, the heterocyclyl may be benzo-fused. The heterocyclyl may be bicyclic, such as, but not limited to, a 5,5 membered ring, such as hexahydrocyclopenta [ c ] pyrrol-2 (1H) -yl ring, or a 5,6 membered bicyclic ring, such as hexahydropyrrolo [1,2-a ] pyrazin-2 (1H) -yl ring. The nitrogen atom containing ring may be partially unsaturated, i.e. it may contain one or more double bonds, such as but not limited to 2, 5-dihydro-1H-pyrrolyl, 4H- [1,3,4] thiadiazinyl, 4, 5-dihydro-oxazolyl or 4H- [1,4] thiazinyl, or it may be benzo-fused, such as but not limited to dihydroisoquinolinyl. According to the invention, the heterocyclic group is non-aromatic.
Advantageous effects
According to a specific example of the present invention, the compound of formula (I) or a tautomer, stereoisomer, hydrate, solvate, pharmaceutically acceptable salt or prodrug of the compound of formula (I) according to the present invention is effective in inhibiting ATX enzyme activity, and exhibits superior hepatic metabolic stability and superior pharmacokinetic properties compared to the control compound.
Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
Detailed Description
The scheme of the present invention will be explained below with reference to examples. It will be appreciated by those skilled in the art that the following examples are illustrative of the present invention and should not be construed as limiting the scope of the invention. The examples are not to be construed as limiting the specific techniques or conditions described in the literature in this field or as per the specifications of the product. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.
Embodiments of the present invention provide compounds of formula (I), tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof, methods and intermediates for preparing compounds of formula (I), tautomers, stereoisomers, hydrates, solvates, pharmaceutically acceptable salts or prodrugs thereof, pharmaceutical compositions, and uses of the compounds and pharmaceutical compositions of the invention in the manufacture of medicaments.
The reaction solvent used in each of the reaction steps described in the present invention is not particularly limited, and any solvent which dissolves the starting materials to some extent and does not inhibit the reaction is included in the present invention. In addition, many similar modifications, equivalent substitutions, or equivalent solvents, combinations of solvents, and different proportions of solvent combinations described herein are considered to be encompassed by the present invention.
The structure of the compounds is determined by Nuclear Magnetic Resonance (NMR) and/or Mass Spectrometry (MS). The unit of NMR shift was 10-6 (ppm). The solvent for NMR measurement is deuterated dimethyl sulfoxide, deuterated chloroform, deuterated methanol, etc., and the internal standard is Tetramethylsilane (TMS).
Liquid chromatography-mass spectrometry (LC-MS) was determined by WatersAcquityH-classUplc-QDA mass spectrometer and monitored using a ACQUITYUPLCBEHC, 2.1 x 50mm,1.7 μm column. Gradient elution conditions were 95-5% solvent A1 and 5-95% solvent B1 at a flow rate of 1.0mL/min, then 95% B1 and 5% A1 were maintained for 0.5min as a percentage of the volume of a certain solvent to the total solvent volume. Wherein solvent A1 is 0.1% formic acid aqueous solution, and solvent B1 is 0.1% formic acid acetonitrile solution. The percentage is the volume percentage of solute in the solution.
Abbreviations for the present invention are defined as follows:
CuI-cuprous iodide
DCM: dichloromethane
DIPEA may also be written as DIEA, diisopropylethylamine, i.e. N, N-diisopropylethylamine
DMF N, N-dimethylformamide
DMSO-dimethyl sulfoxide
Et3 N triethylamine
M: mol/L, for example n-butyllithium (14.56 mL,29.1mmol,2.5M in n-hexane) represents an n-hexane solution of n-butyllithium at a molar concentration of 2.5mol/L
N: equivalent concentration, e.g. 2N hydrochloric acid means 2mol/L hydrochloric acid solution
NADPH reduced coenzyme II
NaH sodium hydrogen, sodium hydride
NMM N-methylmorpholine
NMP N-methylpyrrolidone
Pd/C palladium on carbon
PPh3 triphenylphosphine
SFC supercritical fluid chromatography
T3P-propylphosphoric tricyclic anhydride, i.e. 2,4, 6-tripropyl-1,3,5,2,4,6-trioxatriphosphine-2, 4, 6-trioxide or 1-propylphosphoric anhydride
THF tetrahydrofuran
IC50 half inhibition concentration, which means the concentration at which half of the maximum inhibition effect is achieved.
Unless indicated to the contrary, the compounds exemplified herein are named and numbered using ChemBioDraw Ultra 13.0.0.
Preparation example 1 Synthesis of intermediate B
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (intermediate B)
The synthetic route for the target compound intermediate B is shown below:
first step Synthesis of methyl 2- (butyl-3-yn-1-yloxy) acetate (Compound B-2)
Raw material 3-butynol (compound B-1) (2.8 g,0.4 mol) was added to dry tetrahydrofuran (100 mL), cooled to 0℃and 60% sodium hydrogen (2.4 g,0.6 mmol) was added thereto, stirred at 0℃for 0.5h, raw material methyl 2-bromoacetate (7.3 g,0.48 mmol) was added to the reaction solution, and naturally warmed to room temperature and stirred for 16h. Water (100 mL) was added, extracted with ethyl acetate (50 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =5:1) to give compound B-2 as a colorless liquid (2.24 g, 39% yield).
LC-MSm/z:143.0[M+H]+。
Second step Synthesis of methyl 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetate (Compound B-3)
Compound B-2 (2.24 g,15.7 mmol) was added to N, N-dimethylformamide (15 mL) and methanol (4 mL) at room temperature, and azido trimethylsilane (2.72 g,23.6 mmol), cuprous iodide (356 mg,1.9 mmol) was added under nitrogen, heated to 110℃and stirred for 16h. Water (150 mL) was added, extracted with ethyl acetate (20 mL. Times.3), and the aqueous phase was lyophilized to give compound B-3 as a brown liquid (2.5 g, 85.6% yield).
LC-MSm/z:186.1[M+H]+。
Third step, synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (intermediate B)
Compound B-3 (2.20 g,11.9 mmol) was added to a mixed solution of water (10 mL), tetrahydrofuran (10 mL) and methanol (10 mL) at room temperature, followed by addition of sodium hydroxide (1.9 g,47.6 mmol) and stirring at room temperature for 16h. The reaction solution was concentrated to 15mL, ph=1 to 2 was adjusted, and the aqueous phase was lyophilized to give intermediate B as a yellow liquid (2.1 g, yield 100%).
LC-MSm/z:172.0[M+H]+。
Comparative example 1 control compound and preparation thereof
Reference is made to the synthesis of the control compound from patent application WO2014110000A 1.
The control compounds of the test examples below all refer to the compounds described in comparative example 1.
EXAMPLE 1 Synthesis of Compound 1
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorophenoyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 1)
The synthetic route for target compound 1 is shown below:
First step Synthesis of tert-butyl 2- ((3, 5-dichlorophenyl ethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 1C)
2-Chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (0.75 g,2.93 mmol) and 2- (3, 5-dichlorophenyl) ethylamine (compound 1A) (0.557 g,2.93 mmol), N, N-diisopropylethylamine (1.025 mL,5.87 mmol) were dissolved in N-methylpyrrolidone (5 mL) and stirred at 80℃for 16H. Cooled to room temperature, diluted with distilled water (10 mL), extracted with ethyl acetate (10 ml×3), the organic phases were combined, washed with saturated brine (10 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) =10:1 to 1:1) to give compound 1C (750 mg, yield 62.5%) as a yellow solid.
LC-MSm/z:409.1[M+H]+。
Second step Synthesis of N- (3, 5-dichlorophenyl ethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 1D)
A4M solution of hydrogen chloride in ethyl acetate (10 mL) was added to a round bottom flask containing compound 1C (940 mg, 2.294 mmol) and stirred at room temperature for 1h. After the reaction was completed, the crude compound 1D was concentrated to give a white solid (1.0 g, yield 114%). The crude product was used directly in the next step without purification.
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorophenoyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 1)
O-benzotriazol-N, N, N ', N' -tetramethylurea tetrafluoroboric acid (84 mg,0.262 mmol) was added to N, N-dimethylformamide (5 mL) containing 2- (2- (1H-1, 2, 3-triazol-5-yl) ethoxy) acetic acid (compound 1E), trifluoroacetic acid (37.3 mg,0.131 mmol), N, N-diisopropylethylamine (0.114 mL, 0.254 mmol) at 0 ℃. After stirring for 5 minutes, compound 1D (50 mg,0.131 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 16 hours. Distilled water (20 mL) was added to dilute, extraction was performed with ethyl acetate (20 mL. Times.3), the organic phases were combined, washed with saturated brine (10 mL. Times.2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by preparative chromatography to give compound 1 (4.8 mg, yield 7.93%).
1H NMR(400MHz,MeOD)δ8.22(d,1H),7.73–7.59(m,1H),7.28–7.18(m,3H),4.64(d,2H),4.56(d,2H),4.30(s,2H),3.85(t,2H),3.65-3.61(m,2H),3.07(t,2H),2.89(t,2H).
LC-MSm/z:462.3[M+H]+。
EXAMPLE 2 Synthesis of Compound 2
Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-bis (trifluoromethyl) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 2)
The synthetic route for compound 2 is shown below:
first step Synthesis of tert-butyl 2- ((3, 5-bis (trifluoromethyl) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 2C)
2-Chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 2B) (0.145 g,0.567 mmol) and 2- (3, 5-bis (trifluoromethyl) phenyl) ethylamine hydrochloride (compound 2A) (200 mg,0.68 mmol), diisopropylethylamine (0.294 mL, 1.709 mmol) were dissolved in N-methylpyrrolidone (5 mL) and stirred at 80℃for 16H. Cooled to room temperature, diluted with distilled water (10 mL), extracted with ethyl acetate (10 ml×3), combined with the organic phases, washed with saturated brine (10 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue purified by silica gel column (petroleum ether: ethyl acetate (V/V) =10:1 to 1:1) to give 2- ((3, 5-bis (trifluoromethyl) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 2C) (150 mg, yield 55.5%) as a yellow solid.
LC-MSm/z:477.3[M+H]+。
Second step Synthesis of (N- (3, 5-bis (trifluoromethyl) phenethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 2D)
A4M solution of 1, 4-dioxane (5 mL,20.00 mmol) in hydrogen chloride was added to a round bottom flask containing tert-butyl 2- ((3, 5-bis (trifluoromethyl) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 2C) (150 mg,0.315 mmol) and stirred at room temperature for 3H. Concentration gave (N- (3, 5-bis (trifluoromethyl) phenethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 2D) (145 mg) as a pale yellow solid.
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-bis (trifluoromethyl) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 2)
2,4, 6-Tripropyl-1,3,5,2,4,6-trioxytriphosphate-2, 4, 6-amine dihydrochloride (compound 2D) (145 mg,0.323 mmol) in N, N-dimethylformamide was added dropwise to a solution containing 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (intermediate B) (100 mg,0.578 mmol), triethylamine (0.537 mL,3.85 mmol), (N- (3, 5-bis (trifluoromethyl) phenethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 2D) (145 mg,0.323 mmol) in DMF (3 mL) at 0℃after completion of the reaction, distilled water (30 mL) was added for dilution, the organic phase was combined with saturated brine (10 mL. Times.2), the organic phase was separated, and the sodium sulfate was dried, and the 1- (2, 4-triazol-2-yl) 2-amino-pyrimidine-2-dihydrochloride (compound 2D) (145 mg,0.323 mmol) was isolated, and the anhydrous ethyl-2- (1H-2, 4-triazol-2-yl) was prepared by filtration.
1H NMR(400MHz,MeOD)δ8.24(d,1H),7.94-7.89(m,3H),7.65(s,1H),7.39(s,1H),4.57(s,1H),4.52-4.48(m,2H),4.37(s,1H),4.30(s,2H),3.75(t,2H),3.61-3.59(m,2H),3.09(t,2H),2.91-2.89(m,2H).
LC-MSm/z:530.3[M+H]+。
EXAMPLE 3 Synthesis of Compound 3
2- (2- (1-H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- (phenethylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 3)
The synthetic route for compound 3 is shown below:
first step Synthesis of tert-butyl 2- (phenethylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 3C)
In a single vial was added tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 3A) (1 g,3.92 mmol), 2-phenylethan-1-amine (compound 3B) (0.47 g,3.92 mmol), diisopropylethylamine (1.01 g,7.84 mmol), N-methylpyrrolidone (10 mL) and stirred at 80℃for 16 hours. Cooled to room temperature, diluted with water (10 mL), extracted with ethyl acetate (50 ml×3), the organic phases were combined, washed with saturated brine (50 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) =10:1 to 1:1) to give 2- (phenethylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 3C) (814 mg, yield 61%).
LC-MSm/z:340.4[M+H]+。
Second step, synthesis of N-phenethyl-6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 3D)
In a single vial was added 1, 4-dioxane solution of hydrogen chloride (2.5M, 20 mL), tert-butyl 2- (phenethylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 3C) (814 mg,2.39 mmol), and stirred at room temperature for 1 hour. Filtration and vacuum drying at 50 ℃ afforded N-phenethyl-6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 3D) as a white solid (590 mg, 79% yield).
Third step Synthesis of 2- (2- (1-H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- (phenethylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 3)
N-phenethyl-6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 3D) (100 mg,0.32 mmol), 2- (2- (1H-1, 2, 3-triazol) -4 yl) ethoxy) acetic acid (intermediate B) (112.5 mg,0.65 mmol), diisopropylethylamine (279 mg,2.16 mmol), N, N-dimethylformamide (2 mL) dissolved at room temperature, cooled to 0 ℃, 1-propylphosphoric anhydride (343 mg,0.54mmol,50% N, N-dimethylformamide solution) was added dropwise, the reaction was completed at room temperature overnight after the addition, LCMS showed complete reaction of the starting materials, water (10 mL) was added for dilution, dichloromethane (50 mL. Times.2) was used for extraction, the organic phase was washed with saturated brine (50 mL. Times.2) and separated, the organic phase was dried over sodium sulfate, filtered and concentrated to give 2- (1, 3-triazol-2- (3, 4-amino) -2- (3, 4-triazol) -2-yl) acetic acid hydrochloride (2, 2-dimethyl formamide solution) was prepared by chromatography.
1HNMR(400MHz,CDCl3)δ8.22(d,1H),7.52(s,1H),7.28-7.33(m,2H),7.19-7.23(m,3H),5.25-5.30(m,1H),4.50-4.74(m,4H),4.26-4.28(d,2H),3.92(t,2H),3.69(m,2H),3.07(t,2H),2.90(t,2H).
LC-MSm/z:394.3[M+H]+。
EXAMPLE 4 Synthesis of Compound 4
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 4)
The synthetic route for target compound 4 is shown below:
First step Synthesis of tert-butyl 2- ((4- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carbonate (Compound 4C)
Tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 4A) (598 mg, 2.399 mmol) and 2- (4- (trifluoromethoxy) phenyl) ethyl-1-amine (compound 4B) (320 mg,1.560 mmol), DIEA (806 mg,6.24 mmol) were dissolved in N-methylpyrrolidone (6 mL) and stirred at 105℃for 18 hours. The reaction mixture was added with water (80 mL) and extracted with ethyl acetate (50 mL. Times.3), the organic phases were combined and concentrated. The residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) =3:1) to give compound 4C (467 mg, yield 70.5%) as a white solid.
LC-MSm/z:425.3[M+H]+。
Second step Synthesis of N- (4- (trifluoromethoxy) phenethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 4D)
To compound 4C (700 mg,1.649 mmol) was added a dioxane solution (50 mL) of 2.4N hydrogen chloride, stirred at normal temperature for 1.5 hours, concentrated, and the residue was slurried with methyl tertiary ether to give compound 4D (650 mg, yield 99.4%) as a light brown solid.
LC-MSm/z:325.2[M+H]+。
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 4)
A solution of 1-propanephosphonic acid cyclic anhydride (2.06 g,6.48mmol,50% DMF solution) was added dropwise to a solution of N- (4- (trifluoromethoxy) phenethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 4D) (700 mg,1.77 mmol), triethylamine (1.747 g,17.27 mmol), 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (intermediate B) (739 mg,4.32 mmol) in dichloromethane (20 mL) at 0℃and stirred at room temperature for 18 hours, and TLC monitored the reaction. 30mL of dichloromethane and water (30 mL. Times.3) were added, the organic phase was concentrated, and the residue was purified by a silica gel column (dichloromethane: methanol (V/V) =30:1 to 8 to 1) to give a crude product (450 mg), which was recrystallized from an ethyl acetate/n-heptane system to give compound 4 (334.5 mg, yield 39.6%).
1HNMR(400MHz,CDCl3)δ8.23(d,1H),7.53(s,1H),7.26-7.20(m,2H),7.15(d,2H),5.29(d,1H),4.75(s,1H),4.64(d,2H),4.51(s,1H),4.28(d,2H),3.93(t,2H),3.69(dd,2H),3.08(t,2H),2.92(t,2H).
LC-MSm/z:478.3[M+H]+。
EXAMPLE 5 Synthesis of Compound 5
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 5)
The synthetic route for target compound 5 is shown below:
First step Synthesis of tert-butyl 3- (trifluoromethoxy) phenethylcarbamate (Compound 5B)
2- (3- (Trifluoromethoxy) phenyl) acetonitrile (compound 5A) (1 g,4.92 mmol) was dissolved in methanol (25 mL), nickel chloride hexahydrate (0.117 g,0.492 mmol) was added, the reaction solution was stirred and cooled to 0℃and sodium borohydride (1.862 g,49.2 mmol) was added in portions, and the reaction was continued at room temperature for 16h. Di-tert-butyl dicarbonate (1.611 g,7.38 mmol) was added and the reaction was continued at room temperature for 2h. The reaction mixture was quenched with water (150 mL), extracted with ethyl acetate (50 ml×3), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) =100:1 to 20:1) to give t-butyl 3- (trifluoromethoxy) phenethylcarbamate (compound 5B) as a yellow oil (1.1 g, yield 73.2%).
LC-MSm/z:206.3[M-Boc+H]+。
Second step Synthesis of 2- (3- (trifluoromethoxy) phenyl) ethylamine hydrochloride (Compound 5C)
To t-butyl 3- (trifluoromethoxy) phenethylcarbamate (compound 5B) (0.997 g,3.27 mmol) was added a dioxane solution (5 mL) of 4M hydrogen chloride, and the mixture was stirred at room temperature for 30min. The solvent was removed under reduced pressure to give compound 5C as a yellow solid, and the crude product was used in the next reaction without purification.
Third step Synthesis of tert-butyl 2- ((3- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylate (Compound 5D)
To the crude compound 5C obtained in the second step was added N-methylpyrrolidone (5 mL), tert-butyl 2-amino-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (0.5 g,1.955 mmol), N, N-diisopropylethylamine (1.264 g,9.78 mmol), and the mixture was allowed to react at 100℃for 16H. Cooled to room temperature, water (50 mL) was added, ethyl acetate was extracted (50 ml×3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =2:1) to give tert-butyl 2- ((3- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylate (compound 5D) as a yellow solid (640 mg, yield 77%).
LC-MSm/z:425.4[M+H]+。
Fourth step Synthesis of N- (3- (trifluoromethoxy) phenethyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (Compound 5E)
To tert-butyl 2- ((3- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylate (compound 5D) (640 mg,1.508 mmol) was added a 2.4M solution of hydrogen chloride in dioxane (10 mL) and the reaction stirred at room temperature for 30min. The solvent was removed under reduced pressure to give compound 5E as a yellow viscous solid, and the crude product was used in the next reaction without purification.
Fifth step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 5)
To the crude product of N- (3- (trifluoromethoxy) phenethyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 5E) as a yellow viscous solid obtained in the fourth step was added N, N-dimethylformamide (5 mL), 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (intermediate B) (387 mg,2.262 mmol), triethylamine (1.526 g,15.08 mmol), the reaction solution was cooled to about 0 ℃, and 2,4, 6-tripropyl-1,3,5,2,4,6-trioxytriphosphate-2, 4, 6-trioxide (1.152 g, 1.81mmol, 50% N, N-dimethylformamide solution) was added dropwise, and the mixture was reacted at room temperature for 4H. Water (25 mL) was added to the reaction solution, dichloromethane extraction (15 mL. Times.3), drying the organic phase over anhydrous sodium sulfate, filtration, concentration to dryness, and separation and purification of the residue on a silica gel column (dichloromethane: methanol (V/V) =100:1 to 20:1) gave the compound 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (trifluoromethoxy) phenethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 5) (200 mg, yield 27.8%).
1H NMR(400MHz,DMSO-d6)：δ8.24(d,1H),7.64(bs,1H),7.41-7.13(m,6H),4.55-4.38(m,4H),4.18-4.16(m,2H),3.74-3.71(m,2H),3.51-3.46(m,2H),2.92～2.84(m,4H).
LC-MSm/z:478.3[M+H]+。
EXAMPLE 6 Synthesis of Compound 6
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 6)
The synthetic route for target compound 6 is shown below:
First step Synthesis of tert-butyl-2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid ethyl ester (Compound 6C)
To NMP (10 mL) was added at room temperature tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 6A) (1 g,3.9 mmol), followed by DIEA (3.9 g,30 mmol), (3, 5-dichlorophenyl) methylamine (compound 6B) (1.56 g,9 mmol). The reaction was warmed to 90 ℃, stirred for 8 hours, cooled to room temperature, added with water (10 mL) under ice bath conditions, extracted with ethyl acetate (15 ml×2), the organic phase washed with water (10 mL) and concentrated to give the crude product. The crude product was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =10:1) to give tert-butyl-2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid ethyl ester (compound 6C) (1.4 g, 91% yield).
LC-MSm/z:395.19[M+H]+。
Second step Synthesis of N- (3, 5-dichlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 6D)
Tert-butyl-2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid ethyl ester (1.4 g,3.5 mmol) was added to dioxane solvent of hydrogen chloride (10 mL) under ice bath conditions, and the temperature was raised to room temperature and stirred for 2 hours. The product N- (3, 5-dichlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine hydrochloride (compound 6D) (1 g, 77.8% yield) was obtained by concentration and used directly in the next reaction.
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 6)
Compound 6D (300 mg,0.82 mmol) was added to compound 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (intermediate B) (209 mg,1.22 mmol), DIEA (6 mL), and 1-propanephosphonic acid cyclic anhydride (122 mg,0.38mmol,50% in DMF) with stirring at room temperature, and stirred overnight at room temperature. Water (10 mL) was added and extracted with ethyl acetate (10 mL. Times.2). The organic phases were combined and concentrated to give compound 6 (22 mg, yield 6.0%).
1H NMR(400MHz,CD3OD)δ8.23(d,1H),7.66(s,1H),7.26(d,3H),4.68-4.49(m,6H),4.27(d,2H),3.83(t,2H),3.04(t,2H).
LC-MSm/z:448.00[M+H]+。
EXAMPLE 7 Synthesis of Compound 7
2- (2- (1H-imidazol-1-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 7)
The synthetic route for target compound 7 is shown below:
first step Synthesis of methyl 3-imidazolyl-1-yl propionate (Compound 7B)
Imidazole (30 g,0.44 mol) was added to methyl acrylate (45.5 g,0.53 mol) and potassium phosphate (23.3 g,0.11 mol) at room temperature. Stirring at room temperature for 3 hours, TLC detection reaction was complete, silica gel filtration and concentration gave methyl 3-imidazol-1-yl propionate (34.2 g, yield 50.3%).
Second step Synthesis of 2- (1H-imidazol-1-yl) ethan-1-ol (Compound 7C)
Lithium aluminum hydride (4.04 g,0.11 mol) was added to dry tetrahydrofuran (130 mL) at room temperature and cooled to-thirty degrees Celsius with stirring. At this temperature, methyl 3-imidazolyl-1-yl propionate (13 g,0.08 mol) in tetrahydrofuran (20 mL) was slowly added dropwise, followed by 30 minutes, then warmed to room temperature, stirred for 30 minutes, quenched by dropwise addition of 15% sodium hydroxide (10 mL), extracted with ethyl acetate (30 mL. Times.2), and the organic phases combined. The crude product was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =10:1) to give 2- (1H-imidazol-1-yl) ethan-1-ol (3 g, 24.3% yield).
Third step Synthesis of tert-butyl 2- (2- (1H-imidazol-1-yl) ethoxy) acetate (Compound 7D)
Sodium hydrogen (0.86 g,21.5mmol, 60%) was added to tetrahydrofuran (20 mL) with ice and stirring for 20min, then the compound 2- (1H-imidazol-1-yl) ethan-1-ol (2 g,17.9 mmol) was added, stirring was continued for 30min, tert-butyl bromoacetate (6.99 g,35.8 mmol) was added with ice and stirring was continued overnight at room temperature. Water (10 mL) was added, ethyl acetate (20 mL. Times.2) was extracted twice, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give the crude product, which was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =10:1) to give tert-butyl 2- (2- (1H-imidazol-1-yl) ethoxy) acetate (compound 7D) (3 g, yield 61.7%).
LC-MS m/z:227.15[M+H]+。
Fourth step Synthesis of 2- (2- (1H-imidazol-1-yl) ethoxy) acetic acid (Compound 7E)
The compound tert-butyl 2- (2- (1H-imidazol-1-yl) ethoxy) acetate (3 g,13.2 mmol) was added to 6mL of dioxane hydrochloride solution at room temperature and stirred for 5 hours. The product 2- (2- (1H-imidazol-1-yl) ethoxy) acetic acid (1.6 g, 71.3% yield) was concentrated. Directly used in the next reaction.
Fifth step Synthesis of 2- (2- (1H-imidazol-1-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 7)
Raw material 2- (2- (1- (1H-imidazol-1-yl) ethoxy) acetic acid (121 mg,0.71 mmol) was added to dichloromethane (1 mL) at room temperature, DIEA (1 mL) was added, and N- (3, 5-dichlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 6D) (150 mg,0.41 mmol) and 1-propanephosphonic acid cyclic anhydride (71 mg,0.22mmol,50% in DMF) were added separately under ice-bath conditions and the reaction was warmed to room temperature and stirred for 3 hours.
1H NMR(400MHz,CD3OD)δ8.23(d,1H),7.99(s,1H),7.28(t,4H),7.04(s,1H),4.62-4.50(m,6H),4.33-4.22(m,4H),3.89-3.84(m,2H).
LC-MSm/z:224.21[M/2+H]+。
EXAMPLE 8 Synthesis of Compound 8
(S) -2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) propan-1-one (compound 8)
The synthetic route for compound 8 is shown below:
first step Synthesis of (S) -2- (but-3-yn-1-yloxy) propionic acid (Compound 8C)
To a solution of sodium hydride (2.283 g,57.1mmol, 60%) in anhydrous DMF (20 mL) was added but-3-yn-1-ol (2 g,28.5 mmol), and the mixture was stirred at 0℃for 30min, followed by (R) -2-bromopropionic acid (4.37 g,28.5 mmol). The mixture was stirred at room temperature for 16 hours, after the completion of the reaction, water (200 mL) was added, extraction was performed with ethyl acetate (20 ml×2), ph=1-2 was adjusted by adding 1N diluted hydrochloric acid to the aqueous phase, extraction was performed three times with ethyl acetate/tetrahydrofuran (10:1), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated to give (S) -2- (but-3-yn-1-yloxy) propionic acid (compound 8C) (2.20 g, yield 61%) as a yellow oil.
Second step Synthesis of (S) -2- (but-3-yn-1-yloxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidin-6-yl) propan-1-one (Compound 8D)
To a solution of N- (3, 5-dichlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (0.1 g, 0.399 mmol) in DMF (3 mL) was added successively (S) -2- (but-3-yn-1-yloxy) propionic acid (0.072 g,0.508 mmol) and N-ethyl-N-isopropyl-2-amine (0.219 g,1.69 mmol) at 0deg.C then 1-propylphosphoric anhydride (0.14 g,0.44mmol,50% in DMF) slowly warmed to room temperature and stirred at room temperature for 12 hours. The reaction was quenched with saturated aqueous sodium bicarbonate (10 mL), then diluted with water (50 mL), extracted with ethyl acetate (10 mL. Times.3), and the combined organic layers were concentrated to give the crude product. Purification by prep. plate (DCM: meoh=10:1) afforded (S) -2- (but-3-yn-1-yloxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) propan-1-one (0.1 g, 70.4% yield) as a yellow solid.
Third step (S) -Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) propan-1-one (Compound 8)
The reactant (S) -2- (but-3-yn-1-yloxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) propan-1-one (0.1 g,0.23 mmol), L-ascorbic acid (0.084 g,0.47 mmol) and sodium bicarbonate (0.040 g,0.47 mmol) were added to a mixed solution of DMF (2 mL) and methanol (0.2 mL), followed by addition of copper sulfate pentahydrate (0.014 g,0.055 mmol) and azido trimethylsilane (0.096 g,0.832 mmol) which were stirred at 90℃for 3 hours. The reaction was diluted with water (20 mL), then extracted with DCM (10 ml×3), and the concentrated organic layers were combined and purified by preparative plate to give (S) -2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) propan-1-one (compound 8) (3.1 mg, yield 3.0%).
1H NMR(400MHz,DMSO-d6)δ8.26-8.21(m,1H),7.84-7.81(m,1H),7.49(s,1H),7.41(s,1H),7.30(s,1H),4.70-4.65(m,2H),4.64-4.61(m,2H),4.48-4.40(m,1H),4.24-4.21(m,1H),3.65-3.51(m,3H),2.84-2.74(m,2H),1.23-1.20(m,3H).
LC-MSm/z:462.3[M+H]+。
EXAMPLE 9 Synthesis of Compound 9
2- ((1- (1H-1, 2, 3-triazol-4-yl) propan-2-yl) oxy) -1- (2- (3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 9)
The synthetic route for compound 9 is shown below:
first step Synthesis of 2- ((1- (1H-1, 2, 3-triazol-4-yl) propan-2-yl) oxy) -1- (2- (3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 9)
A50% solution of 1-propylphosphoric anhydride in N, N-dimethylformamide (440 mg,0.692 mmol) was added to a solution of 2- (2- (1H-1, 2, 3-triazol-5-yl) isopropoxy) acetic acid (150 mg, 0.81mmol) in diisopropylethylamine (600 mg,4.65 mmol) in N, N-dimethylformamide (5 mL) at 0 ℃. N- (3, 5-dichlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 6D) (220 mg,0.597 mmol) was added to the reaction solution and stirred at room temperature for 16H. Distilled water (20 mL) was added for dilution, ethyl acetate (20 mL. Times.3) was used for extraction, the organic phases were combined, washed with saturated brine (10 mL. Times.2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by preparative chromatography to give the compound 2- ((1- (1H-1, 2, 3-triazol-4-yl) propan-2-yl) oxy) -1- (2- (3, 5-dichlorobenzyl) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (50 mg, 18.1% yield).
1H NMR(400MHz,DMSO-d6)δ8.24(d,1H),7.84(t,1H),7.61(s,1H),7.41(s,1H),7.30(s,2H),4.54-4.38(m,6H),4.18(q,2H),3.84-3.78(m,1H),2.88-2.78(m,2H),1.09(t,3H).
LC-MSm/z:462.3[M+H]+。
EXAMPLE 10 Synthesis of Compound 10
(R) -2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) propan-1-one (compound 10)
The synthetic route for compound 10 is shown below:
Preparation of example 8 (Compound 8) was analogous to (S) -2-bromopropionic acid instead of (R) -2-bromopropionic acid of example 8, and after the same three steps of reaction, final purification gave (R) -2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-dichlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) propan-1-one (Compound 10) (26.0 mg,0.056 mmol).
1H NMR(400MHz,DMSO-d6)δ8.26-8.21(m,1H),7.85-7.82(m,1H),7.62(br,1H),7.41(s,1H),7.30-7.27(m,2H),4.70-4.56(m,2H),4.49-4.47(m,2H),4.40-4.39(m,1H),4.27-4.21(m,1H),3.64-3.57(m,2H),3.11-3.09(m,1H),2.88-2.84(m,2H),1.23-1.14(m,3H).
LC-MSm/z:462.3[M+H]+。
EXAMPLE 11 Synthesis of Compound 11
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (difluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 11)
The synthetic route for compound 11 is shown below:
First step Synthesis of tert-butyl 2- ((3- (difluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 11C)
2-Chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 11B) (1 g,3.92 mmol) and (3-difluoromethoxy) phenyl) methylamine (compound 11A) (1.35 g,7.80 mmol), diisopropylethylamine (2.5 g,19.40 mmol) were dissolved in N-methylpyrrolidone (10 mL) and stirred at 100℃for 16H. Cooled to room temperature, diluted with distilled water (20 mL), extracted with ethyl acetate (20 ml×3), combined with the organic phase, washed with saturated brine (20 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue purified by silica gel column (petroleum ether: ethyl acetate (V/V) =10:1 to 1:1) to give tert-butyl 2- ((3- (difluoromethoxy) benzyl) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 11C) (1.2 g, yield 77.9%).
Second step Synthesis of N- (3- (difluoromethoxy) benzyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 11D)
Tert-butyl 2- ((3- (difluoromethoxy) benzyl) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylate (compound 11C) (1.2 g,3.06 mmol) was added to a solution of 100mL of hydrogen chloride in 1, 4-dioxane at room temperature, stirred for 8 hours at room temperature, after completion of the monitoring reaction, filtered and the solid dried to give N- (3- (difluoromethoxy) benzyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidine-2-amine dihydrochloride (compound 11D) (950 mg, yield 85.3%).
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (difluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 11)
1-Propylphosphoric anhydride (326 mg,1.025mmol,50% N, N-dimethylformamide solution) was added to a solution of 2- (2- (1H-1, 2, 3-triazol-5-yl) ethoxy) acetic acid (175 mg,1.023 mmol) and diisopropylethylamine (530 mg,4.109 mmol) in N, N-dimethylformamide (2 mL) at 0 ℃. N- (3- (difluoromethoxy) benzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (200 mg,0.55 mmol) was added to the reaction solution, and the mixture was stirred at room temperature for 16H. Distilled water (5 mL) was added for dilution, ethyl acetate (5 mL. Times.3) was used for extraction, the organic phases were combined, washed with saturated brine (2 mL. Times.2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by preparative chromatography to give 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (difluoromethoxy) benzyl) -amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 11) (160 mg, 65.3% yield).
1H NMR(400MHz,CDCl3)δ8.25(d,1H),7.53(s,1H),7.32(t,1H),7.25(s,1H),7.19(d,1H),7.11(s,1H),7.02(d,1H),6.50(td,1H),5.69(brs,1H),4.75(s,1H),4.68-4.63(m,4H),4.51(s,1H),4.27(d,2H),3.92(t,2H),3.08(t,2H).
LC-MSm/z:446.36[M+H]+。
EXAMPLE 12 Synthesis of Compound 12
5- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) isophthalonitrile (compound 12)
The synthetic route for compound 12 is shown below:
First step Synthesis of tert-butyl 2- ((3, 5-dicyanobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 12C)
The starting material, tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 12B) (100 mg,0.39 mmol), was added to DMF (3 mL), 3, 5-dicyanobenzylamine (92 mg,0.59 mmol) and cesium carbonate (255 mg,0.78 mmol) were added, heated to 85℃and stirred for 16H. Filtration, addition of water (30 mL), extraction with ethyl acetate (20 ml×3), combining the organic phases, drying over anhydrous sodium sulfate, filtration, concentration, and separation and purification of the residue on a silica gel column (petroleum ether: ethyl acetate (V/V) =4:1) afforded tert-butyl 2- ((3, 5-dicyanobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 12C) (17 mg, 11.5%) as a yellow solid.
LC-MSm/z:377.1[M+H]+。
Second step Synthesis of 5- (((6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-yl) amino) methyl) isophthalonitrile dihydrochloride (Compound 12D)
Raw material 2- ((3, 5-dicyanobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (500 mg,1.33 mmol) was added to dichloromethane (2 mL), trifluoroacetic acid (1 mL) was added and stirred for 4H at room temperature. Ethyl acetate (20 mL) was added, washed with saturated sodium bicarbonate solution (20 ml×3), the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =10:1), concentrated to give 5- (((6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-yl) amino) methyl) m-phthalonitrile dihydrochloride (compound 12D) (125 mg, yield 27.0%) as a yellow solid.
LC-MSm/z:277.1[M+H]+。
Third step Synthesis of 5- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) isophthalonitrile (Compound 12)
Raw material 5- (((6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-yl) amino) methyl) isophthalonitrile dihydrochloride (compound 12D) (14 mg,0.04 mmol) was added to DMF (2 mL), 1-propylphosphoric anhydride (40 mg,0.061mmol,50% ethyl acetate solution), N-methylmorpholine (11 mg,0.106 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (10 mg,0.061 mmol) were added and stirred for 15H at room temperature. Water (15 mL) was added, extracted with ethyl acetate (15 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 5- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) isophthalonitrile (compound 12) (2.8 mg, 16.3% yield) as a residue.
1H NMR(400MHz,CD3OD)δ8.30(d,1H),8.06(s,1H),8.00(s,2H),7.72(s,1H),4.69(s,4H),4.60(d,2H),4.32(d,2H),3.87(s,2H),3.10(t,2H).
LC-MSm/z:430.1[M+H]+。
EXAMPLE 13 Synthesis of Compound 13
3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile (compound 13)
The synthetic route for compound 13 is shown below:
first step Synthesis of tert-butyl 2- ((3-cyanobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 13B)
Raw material 3- (aminomethyl) benzonitrile (310 mg,2.35 mmol) was added to DMF (5 mL), tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (500 mg,1.96 mmol) and cesium carbonate (1.27 g,3.92 mmol) were added, heated to 85℃and stirred for 16H. Water (50 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((3-cyanophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 13B) as a yellow solid (180 mg, yield 26.1%).
LC-MSm/z:352.1[M+H]+。
Second step Synthesis of 3- (((6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile dihydrochloride 13C)
Raw material 2- ((3-cyanophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 13B) (180 mg,0.51 mmol) was added to dichloromethane (3 mL) at room temperature, trifluoroacetic acid (3 mL) was added, and stirred at room temperature for 16H. Concentration gave 3- (((6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile dihydrochloride (compound 13C) as a yellow solid (150 mg, 91% yield).
LC-MSm/z:252.1[M+H]+。
Third step Synthesis of 3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile (Compound 13)
Raw material 3- (((6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile dihydrochloride (compound 13C) (50 mg,0.16 mmol) was added to N, N-dimethylformamide (2 mL), 1-propylphosphoric anhydride (146 mg,0.23mmol,50% ethyl acetate solution), N-methylmorpholine (38 mg,0.38 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (40 mg,0.37 mmol) were added and stirred at room temperature for 16H. Water (15 mL) was added, extracted with ethyl acetate (15 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the residue as compound 3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile (compound 13) (10 mg, 15.4% yield).
1H NMR(400MHz,DMSO-d6)δ8.30(d,1H),7.95(s,1H),7.72-7.64(m,4H),7.54(t,1H),4.60-4.43(m,6H),4.22(d,2H),3.78-3.74(m,2H),2.95(t,2H).
LC-MSm/z:405.0[M+H]+。
EXAMPLE 14 Synthesis of Compound 14
3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) -5- (trifluoromethoxy) benzonitrile (compound 14)
The synthetic route for compound 14 is shown below:
First step Synthesis of tert-butyl 2- ((3-bromo-5- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 14B)
Raw material 3-bromo-5- (trifluoromethoxy) benzylamine (0.9 g,3.3 mmol) was added to N-methylpyrrolidone (5 mL), tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (1.2 g,4.95 mmol) and diisopropylethylamine (1.24 g,9.9 mmol) were added, heated to 90℃and stirred for 16H. Water (50 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((3-bromo-5- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 14B) (1.2 g, yield 74.5%) as a yellow liquid.
LC-MSm/z:489.1[M+H]+。
Second step Synthesis of tert-butyl 2- ((3-cyano-5- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 14C)
Raw material 2- ((3-bromo-5- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 14B) (1.4 g,2.86 mmol) was added to N, N-dimethylacetamide (15 mL), zinc cyanide (1.4 g,11.96 mmol) and [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (210 mg,0.29 mmol) were added, and the mixture was heated to 160 ℃ with microwaves under nitrogen and stirred for 1H. Water (50 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((3-cyano-5- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 14C) (0.8 g, yield 64.1%) as a white solid.
LC-MSm/z:436.1[M+H]+。
Third step Synthesis of 3- (((6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-yl) amino) methyl) -5- (trifluoromethoxy) benzonitrile dihydrochloride (Compound 14D)
The starting material, tert-butyl 2- ((3-cyano-5- (trifluoromethoxy) phenyl) -amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 14C) (100 mg,0.23 mmol), was added to ethyl acetate (15 mL), and a solution of 4M hydrogen chloride in 1, 4-dioxane (2 mL) was added and stirred at room temperature for 2H. Concentration gave 3- (((6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-yl) amino) methyl) -5- (trifluoromethoxy) benzonitrile dihydrochloride (compound 14D) (36 mg, yield 38.5%) as a white solid.
LC-MSm/z:336.1[M+H]+。
Fourth step Synthesis of 3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) -5- (trifluoromethoxy) benzonitrile (Compound 14)
Raw material 3- (((6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-yl) amino) methyl) -5- (trifluoromethoxy) benzonitrile dihydrochloride (compound 14D) (220 mg,0.54 mmol) was added to N, N-dimethylformamide (5 mL), 1-propylphosphoric anhydride (706 mg,1.86mmol,50% ethyl acetate solution), N-methylmorpholine (310 mg,3.1 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (210 mg,1.24 mmol) were added and stirred at room temperature for 16H. Water (15 mL) was added, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give the compound 3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) -5- (trifluoromethoxy) benzonitrile (compound 14) (18 mg, yield 6.8%) as a residue.
1H NMR(400MHz,CDCl3)δ8.31(d,1H),7.58(d,2H),7.44(d,2H),5.83(s,1H),4.78-4.66(m,5H),4.54(s,1H),4.31(d,2H),3.95(t,2H)3.10(t,2H).
LC-MSm/z:489.1[M+H]+。
EXAMPLE 15 Synthesis of Compound 15
Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2-benzylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 15)
The synthetic route for compound 15 is shown below:
first step Synthesis of tert-butyl 2-benzylamino-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 15B)
2-Chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (0.5 g,1.96 mmol) and benzylamine (0.25 g,2.35 mmol), diisopropylethylamine (1.025 mL,5.87 mmol) were dissolved in N-methylpyrrolidone (5 mL) and stirred at 80℃for 16 hours. Cooled to room temperature, diluted with distilled water (10 mL), extracted with ethyl acetate (10 ml×3), combined with the organic phase, washed with saturated brine (10 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue purified by silica gel column (petroleum ether: ethyl acetate (V/V) =10:1 to 1:1) to give 2-benzylamino-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (350 mg, yield 54.8%) as a yellow solid.
Second step Synthesis of N-benzyl-6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (Compound 15C)
A solution of hydrogen chloride in 1, 4-dioxane (4M, 10 mL) was added to a round bottom flask containing tert-butyl 2-benzylamino-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carbonate (350 mg,0.922 mmol) and stirred at room temperature for 1H. Concentration gave N-benzyl-6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 15C) as a white solid (270 mg, 98% yield). The crude product was used directly in the next step without purification.
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2-benzylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 15)
A50% solution of 1-propylphosphoric anhydride in N, N-dimethylformamide (871 mg,1.370 mmol) was added to a solution of 2- (2- (1H-1, 2, 3-triazol-5-yl) ethoxy) acetic acid (373 mg,1.310 mmol) and diisopropylethylamine (442 mg,3.426 mmol) in N, N-dimethylformamide (5 mL) at 0 ℃. N-benzyl-6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (270 mg,0.906 mmol) was added to the reaction solution and stirred at room temperature for 16H. Distilled water (20 mL) was added for dilution, ethyl acetate (20 mL. Times.3) was used for extraction, the organic phases were combined, washed with saturated brine (10 mL. Times.2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by preparative chromatography to give 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2-benzylamino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (64 mg, yield 18.6%) as a pale yellow solid.
1H NMR(400MHz,DMSO-d6)δ8.23(d,1H),7.51(s,1H),7.33-7.27(m,5H),5.62(b,1H),4.74(s,1H),4.66-4.61(m,4H),4.49(s,1H),4.26(d,2H),3.91(t,2H),3.07(t,2H).
LC-MSm/z:380.2[M+H]+。
EXAMPLE 16 Synthesis of Compound 16
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-difluorophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 16)
The synthetic route for compound 16 is shown below:
first step Synthesis of tert-butyl 2- ((3, 5-difluorophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 16C)
The starting material, tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (800 mg,3.14 mmol), was added to N, N-dimethylformamide (3 mL), 3, 5-difluorobenzylamine (538 mg,3.76 mmol) and cesium carbonate (2.05 g,6.28 mmol) were added, heated to 90℃and stirred for 16H. Filtration, addition of water (30 mL), extraction with ethyl acetate (20 ml×3), combining the organic phases, drying over anhydrous sodium sulfate, filtration, concentration, and separation and purification of the residue on a silica gel column (petroleum ether: ethyl acetate (V/V) =4:1) gave tert-butyl 2- ((3, 5-difluorophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 16C) (500 mg, 43.9%) as a yellow solid.
LC-MSm/z:363.2[M+H]+。
Second step Synthesis of N- (3, 5-difluorophenyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 16D)
Raw material 2- ((3, 5-difluorophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid (500 mg,1.38 mmol) was added to a solution of 4M hydrogen chloride in 1, 4-dioxane (4 mL) and stirred for 2H at room temperature. Concentration gave N- (3, 5-difluorophenyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 16D) as a yellow solid (360 mg, 72% yield).
LC-MSm/z:263.2[M+H]+。
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-difluorophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 16)
The starting material N- (3, 5-difluorophenyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 16D) (150 mg,0.45 mmol) was added to N, N-dimethylformamide (4 mL) and ethyl acetate (2 mL), 1-propylphosphoric anhydride (544 mg,0.86mmol,50% ethyl acetate solution), N-methylmorpholine (115 mg,1.14 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (132 mg,0.63 mmol) were added at room temperature and stirred for 5H. Water (15 mL) was added, extracted with ethyl acetate (15 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =5:1) to give 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3, 5-difluorophenyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 16) (40 mg, 21.3% yield).
1H NMR(400MHz,DMSO-d6)δ8.31(d,1H),7.91(s,1H),7.70(d,1H),7.08-6.99(m,3H),4.60-4.52(m,5H),4.43(s,1H),4.22(d,2H),3.78-3.74(m,2H),2.94(t,2H).
LC-MSm/z:416.2[M+H]+。
EXAMPLE 17 Synthesis of Compound 17
3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) -5-chlorobenzonitrile (compound 17)
The synthetic route for compound 17 is shown below:
First step Synthesis of 3- (azidomethyl) -5-chlorobenzonitrile (Compound 17B)
The starting material 3-chloro-5- (hydroxymethyl) benzonitrile (2.0 g,11.9 mmol) was added to toluene (60 mL) at room temperature, 1, 8-diazabicyclo undec-7-ene (2.18 g,14.4 mmol) was added under nitrogen and stirred for 16h. Water (90 mL) was added, extracted with ethyl acetate (50 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =10:1) to give 3- (azidomethyl) -5-chlorobenzonitrile (compound 17B) (1.53 g, 66% yield) as a colorless liquid.
LC-MSm/z:193.2[M+H]+。
Second step Synthesis of 3-aminomethyl-5-chlorobenzonitrile (Compound 17C)
Raw material 3- (azidomethyl) -5-chlorobenzonitrile (compound 17B) (1.38 g,7.23 mmol) was added to tetrahydrofuran (30 mL) and water (5 mL), then triphenylphosphine (2.08 g,7.95 mmol) was added and stirred at room temperature for 16h, 1M HCl (20 mL) was added, extracted with ethyl acetate (30 ml×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column separation over silica gel (petroleum ether: ethyl acetate (V/V) =1:1) to afford the title compound as a pale yellow solid 3-aminomethyl-5-chlorobenzonitrile (0.7 g, yield 58%).
LC-MSm/z:167.2[M+H]+。
Third step Synthesis of tert-butyl 2- ((3-chloro-5-cyanobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylate (Compound 17D)
The starting material, tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (550 mg,2.15 mmol), was added to N, N-dimethylformamide (8 mL), followed by 3-aminomethyl-5-chlorobenzonitrile (430 mg,2.59 mmol) and potassium carbonate (742 mg,5.37 mmol), heated to 100℃and stirred for 16H. Filtration, addition of water (30 mL), extraction with ethyl acetate (20 ml×3), combination of organic phases, drying over anhydrous sodium sulfate, filtration, concentration, and separation and purification of the residue on a silica gel column (petroleum ether: ethyl acetate (V/V) =1:1) gave tert-butyl 2- ((3-chloro-5-cyanobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylate (compound 17D) (150 mg, yield 21.7%) as a pale yellow solid.
LC-MSm/z:386.2[M+H]+。
Fourth step Synthesis of 3-chloro-5- (((6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile dihydrochloride (Compound 17E)
The starting material, tert-butyl 2- ((3-chloro-5-cyanobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylate (compound 17D) (150 mg,0.38 mmol), was added to methanol (5 mL) and dichloromethane (5 mL), a solution of 4M hydrogen chloride in 1, 4-dioxane (5 mL) was added and stirred at room temperature for 15H. The reaction solution was concentrated, ph=9 to 10 was adjusted with saturated aqueous sodium carbonate, extracted with ethyl acetate (20 ml×3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to give 3-chloro-5- (((6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile dihydrochloride (compound 17E) (100 mg, yield 73.7%) as a pale yellow solid.
LC-MSm/z:286.2[M+H]+。
Fifth step Synthesis of 3- (((6- (2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) -5-chlorobenzonitrile (Compound 17)
Raw material 3-chloro-5- (((6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) benzonitrile dihydrochloride (compound 17E) (100 mg,0.28 mmol) was added to N, N-dimethylformamide (4 mL), then 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (65 mg,0.42 mmol), 1-propylphosphoric anhydride (225 mg,0.7mmol,50% ethyl acetate solution) and N-methylmorpholine (70 mg,0.7 mmol) were added and stirred at room temperature for 16H. Water (15 mL) was added, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =10:1) to give compound 3- (((6- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-yl) amino) methyl) -5-chlorobenzonitrile (compound 17) (40 mg, yield 32%).
1H NMR(400MHz,DMSO-d6)δ8.31(d,1H),7.92(d,2H),7.72-7.69(m,3H),4.60-4.51(m,5H),4.44(s,1H),4.21(d,2H),3.78-3.74(m,2H),2.95(t,2H).
LC-MSm/z:439.0[M+H]+。
EXAMPLE 18 Synthesis of Compound 18
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 18)
The synthetic route for compound 18 is shown below:
first step Synthesis of tert-butyl 2- ((3- (trifluoromethoxy) benzyl) -1-amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 18B)
Raw material 3-trifluoromethoxybenzylamine (675 mg,3.5 mmol) was added to N, N-dimethylformamide (10 mL), tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (811 mg,3.2 mmol) and cesium carbonate (2.3 g,7 mmol) were added, heated to 90℃and stirred for 16H. Water (50 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((3- (trifluoromethoxy) benzyl) -1-amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 18B) as a yellow solid (380 mg, 29% yield).
LC-MSm/z:411.1[M+H]+。
Second step Synthesis of N- (3- (trifluoromethoxy) benzyl)) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (Compound 18C)
Raw material 2- ((3- (trifluoromethoxy) benzyl) -1-amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 18B) (135 mg,0.37 mmol) was added to dichloromethane (38 mL), and a1, 4-dioxane solution of 4M hydrogen chloride (3 mL) was added and stirred for 2H. Concentration gave N- (3- (trifluoromethoxybenzyl)) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 18C) as a yellow solid (116 mg, 82% yield).
LC-MSm/z:311.1[M+H]+。
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 18)
Raw material N- (3- (trifluoromethoxybenzyl)) 6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 18C) (116 mg,0.30 mmol) was added to N, N-dimethylformamide (4 mL) and ethyl acetate (2 mL), 1-propylphosphoric anhydride (176 mg,0.56mmol,50% ethyl acetate solution), N-methylmorpholine (87 mg,1.85 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (63 mg,0.37 mmol) were added, cooled to 0℃and stirred for 0.5H, warmed naturally to room temperature and stirred for 16H. Water (15 mL) was added, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =5:1) to give compound 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3- (trifluoromethoxybenzyl)) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 18) (107 mg, 77% yield).
1H NMR(400MHz,DMSO-d6)δ8.30(d,1H),7.96-7.91(m,1H),7.70(d,1H),7.46(t,1H),7.35(d,1H),7.28(s,1H),7.22(d,1H),4.60-4.43(m,6H),4.22(d,2H),3.78-3.74(m,2H),2.95(t,2H).
LC-MSm/z:464.1[M+H]+。
EXAMPLE 19 Synthesis of Compound 19
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 19)
The synthetic route for compound 19 is shown below:
first step Synthesis of tert-butyl 2- ((3-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 19B)
Raw material 3-chlorobenzylamine (338 mg,2.4 mmol) was added to N-methylpyrrolidone (10 mL), tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (500 mg,2.0 mmol) and N, N-diisopropylethylamine (774 mg,6 mmol) were added, heated to 90℃and stirred for 16H. Water (50 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((3-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 19B) as a yellow solid (570 mg, 80.8% yield).
LC-MSm/z:361.4[M+H]+。
Second step Synthesis of N- (3- (chlorobenzyl)) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (Compound 19C)
The starting material, tert-butyl 2- ((3-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 19B) (150 mg,0.42 mmol), was added to dichloromethane (10 mL), followed by a 4M solution of hydrogen chloride in 1, 4-dioxane (3 mL) and stirred for 2H. Concentration gave N- (3-chlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 19C) as a yellow solid (124 mg, 89% yield).
LC-MSm/z:261.4[M+H]+。
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 19)
Raw material N- (3-chlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine (compound 19C) (110 mg,0.42 mmol) was added to N, N-dimethylformamide (4 mL) and ethyl acetate (2 mL), 1-propylphosphoric anhydride (200 mg,0.63mmol,50% ethyl acetate solution), N-methylmorpholine (212 mg,2.1 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (72 mg,0.42 mmol) were added, cooled to 0℃and stirred for 0.5H, naturally warmed to room temperature, and stirred for 16H. Water (15 mL) was added, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =5:1) to give the compound 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((3-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 19) (40 mg, yield 23%).
1H NMR(400MHz,DMSO-d6)δ8.30(d,1H),7.90(s,1H),7.69(s,1H),7.34-7.26(m,4H),4.59-4.43(m,6H),4.21(d,2H),3.76(d,2H),2.95(t,2H).
LC-MSm/z:414.2[M+H]+。
EXAMPLE 20 Synthesis of Compound 20
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((1- (3, 5-dichlorophenyl) ethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 20)
The synthetic route for compound 20 is shown below:
first step Synthesis of 1- (3, 5-dichlorophenyl) ethane-1-amine (Compound 20B)
The starting material 3, 5-dichloroacetophenone (400 mg,2.1 mmol) was added to methanol (2 mL) at room temperature, followed by tetraethyltitanate (958 mg,4.2 mmol) and concentrated aqueous ammonia (1.5 mL) and stirred at room temperature for 16h. Sodium borohydride (160 mg,4.2 mmol) was added at room temperature and stirred for 3h. Methanol (2 mL) and water (5 mL) were added, filtration, extraction with ethyl acetate (10 ml×3), and drying of the combined organic phases over anhydrous sodium sulfate, filtration, concentration, and purification of the residue by silica gel column (petroleum ether: ethyl acetate (V/V) =1:4) gave 1- (3, 5-dichlorophenyl) ethane-1-amine (compound 20B) as a yellow liquid (258 mg, yield 64%).
LC-MSm/z:190.1[M+H]+。
Second step Synthesis of tert-butyl 2- ((1- (3, 5-dichlorophenyl) ethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 20C)
Raw material 1- (3, 5-dichlorophenyl) ethane-1-amine (compound 20B) (235 mg,1.2 mmol) was added to N-methylpyrrolidone (3 mL), tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (300 mg,1.2 mmol) and N, N-diisopropylethylamine (457 mg,3.5 mmol) were added, heated to 90 ℃ and stirred for 16H. Water (15 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((1- (3, 5-dichlorophenyl) ethyl-1-amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 20C) (117 mg, 28.8% yield) as a yellow liquid.
LC-MSm/z:409.1[M+H]+。
Third step Synthesis of N- (1- (3, 5-dichlorophenyl) ethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (Compound 20D)
Raw material 2- ((1- (3, 5-dichlorophenyl) ethyl-1-amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 20C) (250 mg,0.61 mmol) was added to a solution of 4M hydrogen chloride in 1, 4-dioxane (4 mL) and stirred for 2H, concentrated to give N- (1- (3, 5-dichlorophenyl) ethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidine-2-amine dihydrochloride (compound 20D) (200 mg, 86% yield) as a yellow solid.
LC-MSm/z:309.1[M+H]+。
Fourth step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((1- (3, 5-dichlorophenyl) ethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 20)
Raw material N- (1- (3, 5-dichlorophenyl) ethyl) -6, 7-dihydro-5H-pyrrolo [3,4-D ] pyrimidin-2-amine dihydrochloride (compound 20D) (200 mg,0.52 mmol) was added to N, N-dimethylformamide (4 mL) and ethyl acetate (2 mL), 1-propylphosphoric anhydride (515 mg,0.8mmol,50% ethyl acetate solution), N-methylmorpholine (293 mg,2.9 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (119 mg,0.7 mmol) were added, cooled to 0℃and stirred for 0.5H, naturally warmed to room temperature and stirred for 16H. Water (15 mL) was added, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =5:1) to give 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((1- (3, 5-dichlorophenyl) ethyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 20) (6 mg, yield 2.5%).
1H NMR(400MHz,DMSO-d6)δ8.25(d,1H),7.88(dd,1H),7.64(s,1H),7.42(dt,3H),5.11-5.04(m,1H),4.50(dd,4H),4.17(d,2H),3.74(tt,2H),2.93(t,2H)1.42(d,3H).
LC-MSm/z:462.1[M+H]+。
EXAMPLE 21 Synthesis of Compound 21
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 21)
The synthetic route for compound 21 is shown below:
First step Synthesis of tert-butyl 2- ((4-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 21B)
Raw material 4-chlorobenzylamine (282 mg,2.0 mmol) was added to N-methylpyrrolidone (10 mL), tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (510 mg,2.0 mmol) and N, N-diisopropylethylamine (1.03 g,8 mmol) were added, heated to 90℃and stirred for 16H. Water (50 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((4-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 21B) (300 mg, yield 41.7%) as a yellow solid.
LC-MSm/z:361.4[M+H]+。
Second step Synthesis of N- (4-chlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (Compound 21C)
The starting material, tert-butyl 2- ((4-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 21B) (105 mg,0.29 mmol), was added to dichloromethane (6 mL), followed by a 4M solution of hydrogen chloride in 1, 4-dioxane (2 mL) and stirred for 2H. Concentration gave N- (4-chlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 21C) as a yellow solid (90 mg, 93% yield).
LC-MSm/z:261.4[M+H]+。
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 21)
Raw material N- (4-chlorobenzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 21C) (140 mg,0.42 mmol) was added to N, N-dimethylformamide (4 mL) and ethyl acetate (2 mL), 1-propylphosphoric anhydride (262 mg,0.82mmol,50% ethyl acetate solution), N-methylmorpholine (278 mg,2.75 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (90 mg,0.55 mmol) were added, cooled to 0℃and stirred for 0.5H, naturally brought to room temperature, and stirred for 16H. Water (15 mL) was added, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =5:1) to give the compound 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4-chlorobenzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 21) (50 mg, 29% yield).
1H NMR(400MHz,DMSO-d6)δ8.28(d,1H),7.86(d,1H),7.69(s,1H),7.36-7.30(m,4H),4.59-4.42(m,6H),4.21(d,2H),3.76(d,2H),2.95(t,2H).
LC-MSm/z:414.2[M+H]+。
EXAMPLE 22 Synthesis of Compound 22
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 22)
The synthetic route for compound 22 is shown below:
First step Synthesis of tert-butyl 2- ((4- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 22B)
Raw material 4-trifluoromethoxybenzylamine (compound 22A) (100 mg,0.52 mmol) was added to N, N-dimethylformamide (3 mL), tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (100 mg,0.45 mmol) and cesium carbonate (255 mg,0.78 mmol) were added, heated to 90 ℃ and stirred for 16H. Water (50 mL) was added, extracted with ethyl acetate (20 mL. Times.3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (petroleum ether: ethyl acetate (V/V) =3:1) to give tert-butyl 2- ((4- (trifluoromethoxy) benzyl) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (compound 22B) (12 mg, yield 7.5%) as a yellow solid.
LC-MSm/z:411.1[M+H]+。
Second step Synthesis of N- (4- (trifluoromethoxy) benzyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (Compound 22C)
Raw material 2- ((4- (trifluoromethoxy) benzyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 22B) (150 mg,0.37 mmol) was added to dichloromethane (8 mL), and a solution of 4M hydrogen chloride in 1, 4-dioxane (3 mL) was added and stirred for 2H. Concentration gave N- (4- (trifluoromethoxybenzyl)) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 22C) as a yellow solid (113 mg, 80% yield).
LC-MSm/z:311.1[M+H]+。
Third step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4- (trifluoromethoxybenzyl)) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 22)
Raw material N- (4- (trifluoromethoxybenzyl)) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 22C) (113 mg,0.29 mmol) was added to N, N-dimethylformamide (4 mL) and ethyl acetate (2 mL), 1-propylphosphoric anhydride (175 mg,0.55mmol,50% ethyl acetate solution), N-methylmorpholine (185 mg,1.82 mmol) and 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (63 mg,0.37 mmol) was added, cooled to 0℃and stirred for 0.5H, warmed naturally to room temperature and stirred for 16H. Water (15 mL) was added, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by column chromatography on silica gel (dichloromethane: methanol (V/V) =5:1) to give 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((4- (trifluoromethoxybenzyl)) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 22) (95 mg, yield 70.7%).
1H NMR(400MHz,DMSO-d6)δ8.31(d,1H),7.97(s,1H),7.71(d,1H),7.44(d,2H),7.31(d,2H),4.60-4.44(m,6H),4.22(d,2H),3.78-3.74(m,2H),2.96(t,2H).
LC-MSm/z:464.1[M+H]+。
EXAMPLE 23 Synthesis of Compound 23, compound 24 and Compound 25
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 23)
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((1S, 2R) -2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidin-6-yl) ethan-1-one (compound 24)
2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((1R, 2S) -2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-D ] pyrimidin-6-yl) ethan-1-one (compound 25)
The synthetic routes for compound 23, compound 24 and compound 25 are shown below:
first step Synthesis of tert-butyl (2-phenylcyclopropyl) carbamate (Compound 23B)
Diphenyl azide phosphate (4.12 g,16.96 mmol) was added to a solution of 2-phenylcyclopropanecarboxylic acid (2.5 g,15.41 mmol), triethylamine (1.716 g,16.96 mmol) in t-butanol (19.42 g,262 mmol) at 0 ℃ and reacted overnight at 90 ℃ under nitrogen after displacement. After cooling to room temperature and TLC monitoring, distilled water (50 mL) was added to dilute, extraction was performed with ethyl acetate (100 ml×3), the organic phases were combined, washed with saturated brine (10 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) =10:1 to 1:1) to give tert-butyl (2-phenylcyclopropyl) carbamate (compound 23B) (2.28 g, yield 63.4%) as a yellow solid.
1H NMR(400MHz,DMSO-d6)δ7.26-7.15(m,3H),7.17-7.11(m,1H),7.10-7.05(m,2H),2.60(bs,1H),1.91-1.86(m,1H),1.38(s,9H),1.13-1.04(m,2H).
Second step Synthesis of 2-phenylcyclopropan-1-amine-2, 2-trifluoroacetic acid (Compound 23C)
Trifluoroacetic acid (11.59 mL,150 mmol) was added to dichloromethane (5 mL) containing tert-butyl (2-phenylcyclopropyl) carbamate (compound 23B) (1.95 g,8.36 mmol) and stirred at room temperature for 3h. The reaction solution was concentrated directly to give 2-phenylcyclopropan-1-amine-2, 2-trifluoro-acetic acid (compound 23C) (2.1 g) as a yellow oil.
Third step Synthesis of tert-butyl 2- ((2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (Compound 23E)
Cesium carbonate (5.27 g,16.18 mmol) was added to a solution of N, N-dimethylformamide (20 mL) containing tert-butyl 2-chloro-5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (1.379 g,5.39 mmol) and 2-phenylcyclopropane-1-amine (compound 23C) (2.0 g,8.09 mmol) and stirred at 90 ℃ for 16 hours. After cooling to room temperature and TLC monitoring, distilled water (100 mL) was added to dilute, extraction was performed with ethyl acetate (100 ml×3), the organic phases were combined, washed with saturated brine (10 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by silica gel column (petroleum ether: ethyl acetate (V/V) =10:1 to 1:1) to give 2- ((2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylic acid tert-butyl ester (compound 23E) (800 mg, yield 42.08%).
LC-MSm/z:353.2[M+H]+。
Fourth step Synthesis of N- (2-phenylcyclopropyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (Compound 23F)
A solution of tert-butyl 2- ((2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidine-6-carboxylate (530 mg,15 mmol) (compound 23E) in 4M hydrogen chloride in 1, 4-dioxane (15 mL,36.0 mmol) was stirred at room temperature for 2H. After completion of the reaction was monitored, concentrated to give N- (2-phenylcyclopropyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 23F) (380 mg, yield 78%) as a yellow solid, which was used directly in the next step.
Fourth step Synthesis of 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (Compound 23)
2,4, 6-Tripropyl-1,3,5,2,4,6-trioxytriphosphate-2, 4, 6-trioxide (888 mg, 1.3995 mmol,50% in N, N-dimethylformamide) was added dropwise to a solution of N- (2-phenylcyclopropyl) -6, 7-dihydro-5H-pyrrolo [3,4-d ] pyrimidin-2-amine dihydrochloride (compound 23F) (378 mg,1.16 mmol), 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) acetic acid (239 mg, 1.3995 mmol) in N, N-diisopropylethylamine (2.030 mL,11.62 mmol) in N, N-dimethylformamide (10 mL) at 0℃and stirred at room temperature for 16H. After completion of LCMS monitoring reaction, distilled water (50 mL) was added to dilute, and dichloromethane (100 ml×3) was used to extract, the organic phases were combined, washed with saturated brine (10 ml×2), separated, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was separated by preparative chromatography to give 2- (2- (1H-1, 2, 3-triazol-4-yl) ethoxy) -1- (2- ((2-phenylcyclopropyl) amino) -5, 7-dihydro-6H-pyrrolo [3,4-d ] pyrimidin-6-yl) ethan-1-one (compound 23) (96.0 mg, 20.4% yield).
1H NMR(400MHz,DMSO-d6)δ8.28(d,1H),7.71–7.69(m,2H),7.26(t,2H),7.15(d,3H),4.59-4.42(m,4H),4.21(d,2H),3.77-3.73(m,2H),2.94-2.89(m,3H),1.96-1.94(m,1H),1.28–1.19(m,2H).
LC-MSm/z:406.4[M+H]+。
Fifth step, synthesis of Compound 24 and Compound 25
Compound 24 and compound 25 were resolved by SFC.
The separation conditions were a column of Cellucoat X4.6 mm I.D.,3um, mobile phase consisting of mobile phase A and mobile phase B, mobile phase A being carbon dioxide, mobile phase B being methanol containing 0.05% diethylamine, gradient elution conditions 40% mobile phase B,60% mobile phase A, flow rate 3mL/min, wavelength 220nm, column temperature 35℃and column pressure 100Bar.
Peak 1:
1H NMR(400MHz,CDCl3)δ8.29(d,1H),7.53(d,1H),7.34–7.28(m,2H),7.23-7.19(m,3H),5.60(d,1H),4.76-4.51(m,4H),4.28(d,2H),3.93-3.90(m,2H),3.09-2.98(m,3H),2.09(s,1H),1.40(t,2H).
LC-MSm/z:406.2[M+H]+。
peak 2:
1H NMR(400MHz,CDCl3)δ8.29(d,1H),7.53(d,1H),7.30(t,2H),7.21(t,3H),5.61(d,1H),4.76-4.52(m,4H),4.28(d,2H),3.92(d,2H),3.11–2.95(m,3H),2.09(s,1H),1.47–1.41(m,1H),1.35-1.34(m,1H).
LC-MSm/z:406.2[M+H]+。
Biological Activity and related Property test cases
Test example 1 Autotaxin (ATX) enzyme Activity inhibition assay
The inhibitory activity of the compounds on Autotaxin enzyme was tested using Autotaxin Inhibitor SCREENING ASSAY KIT (Cayman, 700580). Test compounds were first formulated as 10mM stock solution in DMSO solvent, then diluted with DMSO gradient at 8 concentration points, and then diluted at 8 concentration points with Autotaxin Assay buffer (1X) provided in the kit to 19X working solution of compound (DMSO content 1.9%). Autotaxin ASSAY REAGENT (10×) was taken out and diluted 10-fold with Autotaxin Assay Buffer (1×). Taken out Autotaxin Substrate, added with 1.2: 1.2mL Autotaxin Assay Buffer (1×) for dissolution, mixed evenly and stood at room temperature. In a 96-well plate, 150 mu L Autotaxin Assay Buffer (1×) of diluted 19×compound working solution, 10 mu L Autotaxin ASSAY REAGENT (1×) of diluted 20 mu L of dissolved Autotaxin Substrate are added into each well of each concentration point, uniformly mixed, incubated for 30min at a constant temperature of 37 ℃ through a shaking table in a dark place, the 96-well plate is taken out, the OD405 is read on an enzyme-labeled instrument, experimental results are input into GRAPHPAD PRISM software, and the IC50 of each compound is obtained through fitting calculation.
TABLE 1 results of test Compounds for inhibitory Activity on ATX enzyme Activity
Experimental results show that the compound has good inhibitory activity on ATX enzyme.
Test example 2 stability test of human liver microsomes
The stability test of the human liver microsome is carried out by adopting the co-incubation of the compound and the human liver microsome in vitro. Test compounds were first formulated as a 10mM stock solution in DMSO solvent, followed by dilution of the compounds to 0.5mM using acetonitrile. Human liver microsomes (Corning) were diluted with PBS to a microsome/buffer solution, and 0.5mM of the compound was diluted with the solution to a working solution having a compound concentration of 1.5. Mu.M and a human liver microsome concentration of 0.75mg/ml. The reaction was started by taking a deep well plate, adding 30. Mu.L of working solution per well, then adding 15. Mu.L of pre-warmed 6mM NADPH solution, and incubating at 37 ℃. At 0, 5, 15, 30, 45 minutes of incubation, the reaction was terminated by adding 135 μl acetonitrile to the corresponding wells. After the reaction was terminated with acetonitrile at the last 45 min time point, the deep-well plate was vortexed for 10 minutes (600 rpm/min) and then centrifuged for 15 minutes. Taking the supernatant after centrifugation, adding purified water in a ratio of 1:1, performing LC-MS/MS detection to obtain the ratio of the peak area of the compound to the internal standard peak area at each time point, comparing the ratio of the peak area of the compound at 5, 15, 30 and 45 minutes with the ratio of the peak area at 0 minute, calculating the residual percentage of the compound at each time point, and calculating T1/2 by using Excel.
TABLE 2 results of human liver microsome stability test
Experimental results show that the compound of the invention has better liver metabolism stability, slower metabolism in human body and higher exposure.
Test example 3 pharmacokinetic test
In vivo pharmacokinetic experiments of mice, 6 male mice, 25-30g, were used and fasted overnight. 3 rats were harvested and 10mg/kg of the drug was orally administered by gavage, and blood was collected 15, 30 minutes and 1,2, 4, 8, 24 hours before and after administration. An additional 3 mice were taken, 1mg/kg given by intravenous injection, and blood was collected 5, 15, 30 minutes and 1,2, 4, 8, 24 hours before and after administration. Blood samples were centrifuged at 8000 rpm at 4℃for 6 minutes and plasma was collected and stored at-20 ℃. Plasma at each time point is taken, 3-5 times of acetonitrile solution containing an internal standard is added for mixing, vortex mixing is carried out for 1 minute, 13000 r/min and 4 ℃ are centrifugated for 10 minutes, 3 times of water is added for mixing the supernatant, and a proper amount of mixed solution is taken for LC-MS/MS analysis. The principal pharmacokinetic parameters were analyzed using the WinNonlin 7.0 software non-compartmental model.
TABLE 3 results of in vivo pharmacokinetic experiments in mice
Experimental results show that the compound of the invention shows more excellent pharmacokinetic properties.