Compound, pharmaceutical composition, and method for preparing and using the compound

Abstract

Disclosed are compounds and pharmaceutically acceptable salts thereof that can be used in treating a subject in need thereof. The compounds disclosed herein can be inhibitors of tyrosine- and threonine-specific cdc2 inhibitory kinase (Myt1). Also disclosed are pharmaceutical compositions containing the compounds or pharmaceutically acceptable salts thereof, as well as methods for preparing and using the same.

Description

Technical Field

[0001] The present invention relates to compounds and pharmaceutical compositions, their preparation, and their use in the treatment of diseases or conditions, such as cancer, in particular diseases or conditions that depend on the activity of membrane-bound tyrosine and threonine-specific cdc2 inhibitory kinase (Myt1) (gene name PKMYT1) (e.g., cancers involving CCNE1 amplification / overexpression, or FBXW7 mutant cancers).

Background Art

[0002] DNA is continuously exposed to both endogenous (e.g., replication fork stalling, reactive oxygen species) and exogenous (UV, ionizing radiation, chemicals) damage-causing factors that can cause DNA damage. As a result, cells have established sophisticated mechanisms to counter these harmful events that would otherwise compromise genomic integrity and lead to genomic instability diseases such as cancer. These mechanisms are collectively referred to as the DNA damage response (DDR). One component of the overall DDR is the activation of various checkpoint pathways that regulate specific DNA repair mechanisms throughout the different stages of the cell cycle, which include the G1, S, G2, and mitotic checkpoints. The majority of cancer cells have lost their G1 checkpoint due to p53 mutations and thus rely on the G2 checkpoint to perform the required DNA damage correction before proceeding to mitosis and dividing into two daughter cells.

[0003] There is a need for new anti-cancer treatment approaches, such as those utilizing small molecules, in particular therapies that enable targeted cancer treatment.

Summary of the Invention

[0004] In one aspect, the present invention provides a compound of formula (I):

Chemical formula

[0005] In some embodiments, the compound is an atropisomer of formula (IA):

Chemical formula

[0006] In some embodiments, X is CR 2 . In some embodiments, the compound has the formula (II):

Chemical formula

[0007] In some embodiments, the compound is an atropisomer of formula (IIA):

Chemical formula

[0008] In some embodiments, the compound has the formula (III):

Chemical formula

[0009] In some embodiments, R 7B is hydrogen, optionally substituted C 2A alkyl, or halogen.

[0010] In some embodiments, R 1-6 is optionally substituted C 3 alkyl. In some embodiments, R 1-6 is halogen. In some embodiments, R 3 is optionally substituted C 4 1-6is alkyl. In some embodiments, R 4 is halogen (e.g., chlorine).

[0011] In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is optionally substituted C 1-6 alkyl. R 2 is optionally substituted methyl, or optionally substituted isopropyl. R 2 is halogen.

[0012] In some embodiments, R 1 is hydrogen. In some embodiments, R 1 is halogen. In some embodiments, R 1 is chlorine or bromine. In some embodiments, R 1 is optionally substituted C 1-6 alkyl. In some embodiments, R 1 is optionally substituted methyl, optionally substituted ethyl, optionally substituted isopropyl, or optionally substituted butyl. In some embodiments, R 1 is optionally substituted C 1-9 heteroaryl. In some embodiments, R 1 is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidinyl, pyrazinyl, pyridazinyl or pyrazolyl, and R 1 is optionally substituted with substituents defined for optionally substituted C 1-9 heteroaryl. In some embodiments, R 1 is optionally substituted C 3-8 cycloalkyl. In some embodiments, R 1 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and R 1 is optionally substituted C3-8 It is optionally substituted with a substituent defined for cycloalkyl. In some embodiments, R 1 is optionally substituted C 2-9 heterocyclyl. In some embodiments, R 1 is 1,2,3,6-tetrahydropyridinyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, oxa-aza-spiro[3.3]heptane, or oxa-aza-bicyclo[3.2.1]octane, and R 1 is optionally substituted C 2-9 heterocyclyl optionally substituted with a substituent defined for heterocyclyl. In some embodiments, R 1 is optionally substituted C 3-8 cycloalkyl. In some embodiments, R 1 is optionally substituted cyclohexenyl, or optionally substituted cyclopentenyl. In some embodiments, R 1 is optionally substituted C 6-10 aryl. In some embodiments, R 1 is optionally substituted phenyl.

[0013] In some embodiments, R 1 is -Q-R 7B . In some embodiments, Q is optionally substituted C 2-6 alkynylene. In some embodiments, Q is optionally substituted C 1-6 alkylene. In some embodiments, Q is optionally substituted C 6-10 arylene. In some embodiments, R 7B is optionally substituted C 2-9 heterocyclyl. In some embodiments, R 7B is optionally substituted C 6-10 aryl.

[0014] In some embodiments, R 1is independently selected from the group consisting of methyl, difluoromethyl, trifluoromethyl, fluorine, chlorine, bromine, amino, hydroxyl, cyano, oxo, -C(O)NH2, -C(O)NH(Me), -C(O)N(Me)2, -(CH2) n -C(O)OH, and -(CH2) n -C(O)Ot-Bu and is optionally substituted with one, two or three groups, and n is 0 or 1.

[0015] In some embodiments, R 1 is -N(R 7 )2. In some embodiments, R 1 is diethylamino.

[0016] In some embodiments, R 5 is hydrogen. In some embodiments, R 5 is -N(R 7 )2. In some embodiments, R 5 is -NH2. In some embodiments, R 6 is -C(O)NH(R 8 ). In some embodiments, R 6 is -C(O)NH2. In some embodiments, R 6 is -C(O)NH(Me). In some embodiments, R 6 is -SO2R 7A . In some embodiments, R 6 is -SO2Me.

[0017] In some embodiments, a compound selected from the group consisting of Compounds 1-328 (e.g., Compounds 1-288) and pharmaceutically acceptable salts thereof.

[0018] In another aspect, the present invention provides a pharmaceutical composition comprising a compound disclosed herein or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient. In some embodiments, the composition is isotopically enriched with deuterium.

[0019] In yet another aspect, the present invention provides a method of inhibiting Myt1 in cells in which Myt1 is expressed, the method comprising contacting the cells with a compound disclosed herein.

[0020] In some embodiments, the cells overexpress CCNE1. In some embodiments, the cells are in a subject.

[0021] In yet another aspect, the present invention provides a method of treating a subject in need thereof, the method comprising administering to the subject a compound disclosed herein, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition disclosed herein.

[0022] In some embodiments, the subject is suffering from a disease or condition having symptoms of cell overproliferation and in need of treatment therefor. In some embodiments, the disease or condition is cancer. In some embodiments, the cancer is a cancer in which CCNE1 is overexpressed.

[0023] In yet another aspect, the present invention provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a Myt1 inhibitor, wherein the cancer has been previously identified as a cancer in which CCNE1 is overexpressed.

[0024] In another aspect, the present invention provides a method of treating cancer in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a Myt1 inhibitor, wherein the cancer is a cancer in which CCNE1 is overexpressed.

[0025] In yet another aspect, the present invention provides a method of inducing cell death in cancer cells in which CCNE1 is overexpressed, the method comprising contacting the cells with an effective amount of a Myt1 inhibitor.

[0026] In some embodiments, the cells are in a subject. In some embodiments, the Myt1 inhibitor is a compound disclosed herein or a pharmaceutically acceptable salt thereof. In some embodiments, the cancer in which CCNE1 is overexpressed is uterine cancer, ovarian cancer, breast cancer, gastric cancer, esophageal cancer, lung cancer or endometrial cancer.

[0027] In yet another aspect, the present invention provides a method of treating cancer in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an Myt1 inhibitor, wherein the cancer has been previously identified as a cancer having an inactivating mutation in the FBXW7 gene.

[0028] In another aspect, the present invention provides a method of treating cancer in a subject, comprising administering to a subject in need thereof a therapeutically effective amount of an Myt1 inhibitor, wherein the cancer has an inactivating mutation in the FBXW7 gene.

[0029] In yet another aspect, the present invention provides a method of inducing cell death in FBXW7 mutant cancer cells, comprising contacting the cells with an effective amount of an Myt1 inhibitor.

[0030] In some embodiments, the cells are in a subject. The cancer is uterine cancer, colorectal cancer, breast cancer, lung cancer or esophageal cancer. In some embodiments, the Myt1 inhibitor is a compound disclosed herein or a pharmaceutically acceptable salt thereof.

[0031] Abbreviations Abbreviations and terms commonly used in the fields of organic chemistry, medicinal chemistry, pharmacology, and medicine and well-known to those skilled in the art are used herein. Representative abbreviations and definitions are shown below.

[0032] Ac is acetyl [CH3C(O)-]; Ac2O is acetic anhydride; AcOH is acetic acid; APC is an antigen-presenting cell; aq. is aqueous; 9-BBN is 9-borabicyclo[3.3.1]nonane; BINAP is (2,2'-bis(diphenylphosphino)-1,1'-binaphthyl); Bn is benzyl; BOC is tert-butyloxycarbonyl; CDI is carbonyldiimidazole; DCM is dichloromethane; DIAD is diisopropyl azodicarboxylate; DIBAL is diisobutylaluminum hydride; DIPEA is diisopropylethylamine; DMA is dimethylacetamide; DMAP is 4-dimethylaminopyridine; DMF is N,N-dimethylformamide; DMSO is dimethyl sulfoxide; dppf is 1,1'-bis(diphenylphosphino)ferrocene; EDAC (or EDC) is 1-ethyl-3-[3-(dimethylamino)propyl]-carbodiimide HCl; ESI is electrospray ionization mass spectrometry; Et2O is diethyl ether; Et3N is triethylamine; Et is ethyl; EtOAc is ethyl acetate; EtOH is ethanol; 3-F-Ph is 3-fluorophenyl; HATU is (1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate; HCl is hydrochloric acid; HOBt is 1-hydroxybenzotriazole; HPLC is high performance liquid chromatography; LCMS is HPLC with mass spectrum detection; LiHMDS is lithium bis(trimethylsilyl)amide; LG is a leaving group; M is molarity; mCPBA is meta-chloroperbenzoic acid; mmol is millimole; Me is methyl; MeCN is acetonitrile; MeOH is methanol; Ms is methanesulfonyl; MS is mass spectrometry; N is normal; NaHMDS is sodium hexamethyldisilazide; NaOAc is sodium acetate; NaOtBu is sodium tert-butoxide; NMO is N-methylmorpholine N-oxide; NMP is N-methylpyrrolidinone;NMR is nuclear magnetic resonance spectroscopy; Pd2(dba)3 is tris(dibenzylideneacetone)dipalladium; PdCl2(PPh3)2 is dichlorobis-(triphenylphosphine)palladium; PG represents an unspecified protecting group; Ph is phenyl; PhMe is toluene; PPh3 is triphenylphosphine; PMB is paramethoxybenzyl; rt is room temperature; RBF is a round bottom flask; RuPhos Pd G1 is chloro-(2-dicyclohexylphosphino-2’,6’-diisopropoxy-1,1’-biphenyl)[2-(2-aminoethyl)phenyl]palladium(II); SEM is [2-(trimethylsilyl)ethoxy]methyl; SFC is supercritical fluid chromatography; S; N Ar is nucleophilic aromatic substitution; TBAB is tetrabutylammonium bromide; TBAF is tetrabutylammonium fluoride; TBS is tert-butyldimethylsilyl; tBu is tert-butyl; Tf is triflate; TFA is trifluoroacetic acid; THF is tetrahydrofuran; THP is tetrahydropyran; TLC is thin layer chromatography; TMAD is tetramethylazodicarboxamide; TMS is trimethylsilyl; TPAP is tetrapropylammonium perruthenate; Ts is p-toluenesulfonyl; UPLC is ultra performance liquid chromatography.

[0033] Definitions As used herein, the term "abnormal" refers to being different from normal. When used in the context of an activity, abnormal refers to the activity being greater than or less than the average of a normal control or a normal non-diseased control sample. Abnormal activity may refer to an amount of activity that causes a disease, in which case, when the abnormal activity returns to a normal or non-disease related amount (e.g., by administering a compound described herein or using a method described herein), a reduction in the disease or one or more disease symptoms is obtained.

[0034] As used herein, the term "acyl" represents a group -C(=O)-R, where R represents alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, or heterocyclyl. The acyl may be optionally substituted as described herein for each R group.

[0035] As used herein, the term "adenocarcinoma" represents a malignant tumor resulting from glandular cells that line an organ within an organism. Non - limiting examples of adenocarcinoma include non - small cell lung cancer, prostate cancer, pancreatic cancer, esophageal cancer, and colorectal cancer.

[0036] As used herein, the term "alkanoyl" represents a group in which a hydrogen or an alkyl group is bonded to a parent molecular group through a carbonyl group, and is exemplified by formyl (i.e., carboxaldehyde group), acetyl, propionyl, butyryl, and isobutyryl. An unsubstituted alkanoyl group contains 1 to 7 carbons. The alkanoyl group may be unsubstituted or substituted for the alkyl group as described herein (e.g., optionally substituted C1 - 7 alkanoyl). By adding the terminal "-(o)yl" to another group defined herein, such as aryl, cycloalkyl, and heterocyclyl, "aroyl", "cycloalkanoyl", and "(heterocyclyl)oyl" can be defined. These groups each represent a carbonyl group substituted with aryl, cycloalkyl, or heterocyclyl. Each of "aroyl", "cycloalkanoyl", and "(heterocyclyl)oyl" may be optionally substituted as defined for "aryl", "cycloalkyl", or "heterocyclyl".

[0037] As used herein, the term "alkenyl" refers to a monovalent straight-chain or branched acyclic hydrocarbon group containing one, two, or three carbon-carbon double bonds. Non-limiting examples of alkenyl groups include ethenyl, prop-1-enyl, prop-2-enyl, 1-methylethenyl, but-1-enyl, but-2-enyl, but-3-enyl, 1-methylprop-1-enyl, 2-methylprop-1-enyl, and 1-methylprop-2-enyl. The alkenyl group may be optionally substituted as defined herein for alkyl.

[0038] As used herein, the term "alkenylene" refers to a divalent alkenyl group. Optionally substituted alkenylene is alkenylene that is optionally substituted as described herein for alkenyl.

[0039] As used herein, the term "alkoxy", unless otherwise specified, represents a chemical substituent of the formula -OR, where R is a C 1-6 alkyl group. In some embodiments, the alkyl group can be further substituted as defined herein. The term "alkoxy" can be combined with other terms defined herein, such as aryl, cycloalkyl, or heterocyclyl, to define "arylalkoxy" groups, "cycloalkylalkoxy" groups, and "(heterocyclyl)alkoxy" groups. These groups represent alkoxy groups substituted with aryl, cycloalkyl, or heterocyclyl, respectively. Each of "arylalkoxy", "cycloalkylalkoxy", and "(heterocyclyl)alkoxy" may be optionally substituted as defined herein for each individual moiety.

[0040] As used herein, the term "alkoxyalkyl" refers to a group of the formula -L-O-R, where L is a C 1-6 alkylene and R is a C 1-6represents a chemical substituent that is alkyl. Optionally substituted alkoxyalkyl is optionally substituted alkoxyalkyl as described herein for alkyl.

[0041] As used herein, the term "alkyl" refers to a straight-chain or branched-chain acyclic saturated hydrocarbon group which, when unsubstituted, has from 1 to 12 carbon atoms, unless otherwise specified. In certain preferred embodiments, unsubstituted alkyl has from 1 to 6 carbon atoms. Alkyl groups are exemplified by methyl; ethyl; n-propyl and isopropyl; n-butyl, sec-butyl, isobutyl, and tert-butyl; neopentyl, etc., and may be optionally substituted with one, two, three substituents, or in the case of alkyl groups having two or more carbons, four or more substituents within the valence tolerance, and the substituents are independently selected from the group consisting of amino; alkoxy; aryl; aryloxy; azide; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heterocyclyl; (heterocyclyl)oxy; heteroaryl; hydroxy; nitro; thiol; silyl; cyano; alkylsulfonyl; alkylsulfinyl; alkylsulfenyl; =O; =S; -C(O)R or -SO2R (wherein R is amino); and =NR' (wherein R' is H, alkyl, aryl, or heterocyclyl). Each of the substituents may itself be unsubstituted or may be substituted with unsubstituted substituent(s) as defined herein for each group within the valence tolerance.

[0042] As used herein, the term "alkylene" refers to a divalent alkyl group. Optionally substituted alkylene is optionally substituted alkylene as described herein for alkyl.

[0043] As used herein, the term "alkylamino" refers to the formula -N(R N1 )2 or -NHR N1 (wherein R N1refers to a group having a group (wherein the alkyl group is an alkyl group). The alkyl portion of the alkylamino may be optionally substituted as defined for alkyl. Each optional substituent of the substituted alkylamino may itself be unsubstituted or, within the valence tolerance, may be substituted with an unsubstituted substituent(s) defined herein for each group.

[0044] As used herein, the term "alkylsulfenyl" represents a group of the formula -S-(alkyl). The alkylsulfenyl may be optionally substituted as defined for alkyl.

[0045] As used herein, the term "alkylsulfinyl" represents a group of the formula -S(O)-(alkyl). The alkylsulfinyl may be optionally substituted as defined for alkyl.

[0046] As used herein, the term "alkylsulfonyl" represents a group of the formula -S(O)2-(alkyl). The alkylsulfonyl may be optionally substituted as defined for alkyl.

[0047] As used herein, the term "alkynyl" represents a monovalent straight-chain or branched-chain hydrocarbon group having 2 to 6 carbon atoms and containing at least one carbon-carbon triple bond, exemplified by ethynyl, 1-propynyl, etc. The alkynyl group may be unsubstituted or substituted as defined for alkyl (e.g., optionally substituted alkynyl).

[0048] As used herein, the term "alkynylene" refers to a divalent alkynyl group. Optionally substituted alkynylene is an alkynylene that is optionally substituted as described herein for alkynyl.

[0049] As used herein, the term "amino" represents -N(R N1 )2, and when the amino is unsubstituted, R N1All are H; when the amino group is substituted, each R N1 is independently H, -OH, -NO2, -N(R N2 )2, -SO2OR N2 , -SO2R N2 , -SOR N2 , -C(O)OR N2 , an N-protecting group, alkyl, alkenyl, alkynyl, alkoxy, aryl, arylalkyl, aryloxy, cycloalkyl, cycloalkenyl, heteroalkyl, or heterocyclyl, provided that at least one R N1 is not H, and each R N2 is independently H, alkyl, or aryl. Each of the substituents may itself be unsubstituted or may be substituted with an unsubstituted substituent(s) as defined herein for each group. In some embodiments, the amino group is an unsubstituted amino (i.e., -NH2) or a substituted amino (e.g., NHR N1 ), in which case R N1 is independently -OH, SO2OR N2 , -SO2R N2 , -SOR N2 , -COOR N2 , optionally substituted alkyl, or optionally substituted aryl, and each R N2 can be optionally substituted alkyl or optionally substituted aryl. In some embodiments, the substituted amino can be an alkylamino in which the alkyl group is optionally substituted as described herein for alkyl. In some embodiments, the amino group is -NHR N1 , and R N1 is optionally substituted alkyl.

[0050] As used herein, the term "aryl" refers to a monocyclic, bicyclic, or polycyclic carbocyclic system having one or two aromatic rings. An aryl group may contain 6 to 10 carbon atoms. All atoms within an unsubstituted carbocyclic aryl group are carbon atoms. Non-limiting examples of carbocyclic aryl groups include phenyl, naphthyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indanyl, indenyl, and the like. An aryl group may be unsubstituted or substituted with one, two, three, four, or five substituents independently selected from the group consisting of alkyl; alkenyl; alkynyl; alkoxy; alkylsulfinyl; alkylsulfenyl; alkylsulfonyl; amino; aryl; aryloxy; azide; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heteroalkyl; heterocyclyl; (heterocyclyl)oxy; hydroxy; nitro; thiol; silyl; -(CH2) n -C(O)OR A ;-C(O)R; and -SO2R, wherein R is amino or alkyl, and R A is H or alkyl, and n is 0 or 1. Each of the substituents may itself be unsubstituted or may be substituted with an unsubstituted substituent(s) as defined herein for each group.

[0051] As used herein, the term "arylalkyl" refers to an alkyl group substituted with an aryl group. The aryl moiety and the alkyl moiety may each be optionally substituted as described herein for the individual groups.

[0052] As used herein, the term "arylene" refers to a divalent aryl group. An optionally substituted arylene is an optionally substituted arylene as described herein for aryl.

[0053] As used herein, the term "aryloxy" refers to a chemical substituent of the formula -OR, where R is an aryl group, unless otherwise specified. In aryloxy optionally substituted, the aryl group is optionally substituted as described herein for aryl.

[0054] As used herein, the term "azide" refers to the -N3 group.

[0055] As used herein, the term "cancer" refers to any type of cancer, neoplasm, or malignant tumor found in a mammal (e.g., human).

[0056] As used herein, the term "carbocyclic" refers to an optionally substituted C3-16 monocyclic, bicyclic, or tricyclic structure in which the ring, which can be aromatic or non-aromatic, is formed by carbon atoms. Examples of carbocyclic structures include cycloalkyl groups, cycloalkenyl groups, cycloalkynyl groups, and certain aryl groups.

[0057] As used herein, the term "carbonyl" refers to the -C(O)- group.

[0058] As used herein, the term "carcinoma" refers to a malignant neoplasm composed of epithelial cells that tend to invade surrounding tissues and cause metastasis.

[0059] As used herein, the term "cyano" refers to the -CN group.

[0060] As used interchangeably herein, the terms "CCNE1" and "cyclin E1" refer to cyclin E1 (gene name CCNE1) specific to the G1 / S phase. Cells in which CCNE1 is overexpressed are cells that exhibit higher CCNE1 activity compared to cells in which CCNE1 is normally expressed. For example, CCNE1 overexpressing cells are cells that exhibit at least 3 copy numbers as opposed to diploid normal cells having 2 copies. Cells exhibiting a copy number greater than 3 of CCNE1 are cells in which CCNE1 is overexpressed. CCNE1 overexpression can be measured by determining the expression level of the gene product in the cell (e.g., CCNE1 mRNA transcript count, or CCNE1 protein level).

[0061] As used herein, the term "cycloalkenyl" refers to a non-aromatic carbocyclic group having at least one double bond and 3 to 10 carbons in the ring (e.g., C 3-10 cycloalkenyl), unless otherwise specified. Non-limiting examples of cycloalkenyl include cycloprop-1-enyl, cycloprop-2-enyl, cyclobut-1-enyl, cyclobut-1-enyl, cyclobut-2-enyl, cyclopent-1-enyl, cyclopent-2-enyl, cyclopent-3-enyl, norbornen-1-yl, norbornen-2-yl, norbornen-5-yl, and norbornen-7-yl. The cycloalkenyl group may be unsubstituted or substituted as described for cycloalkyl (e.g., optionally substituted cycloalkenyl).

[0062] As used herein, the term "cycloalkenylalkyl" represents an alkyl group substituted with a cycloalkenyl group as defined herein. The cycloalkenyl moiety and the alkyl moiety may be substituted as described for the individual groups defined herein.

[0063] As used herein, the term "cycloalkenylene" represents a divalent cycloalkenyl group. Optionally substituted cycloalkenylene is optionally substituted cycloalkenylene as described herein for cycloalkyl.

[0064] As used herein, the term "cycloalkoxy", unless otherwise specified, represents a chemical substituent of the formula -OR, where R is a cycloalkyl group. In some embodiments, the cycloalkyl group can be further substituted as defined herein.

[0065] As used herein, the term "cycloalkyl", unless otherwise specified, refers to a cyclic alkyl group having 3 to 10 carbons (e.g., C 3-C10refers to (cycloalkyl). The cycloalkyl group can be monocyclic or bicyclic. The bicyclic cycloalkyl group can be of the bicyclo[p.q.0]alkyl type where each of p and q is independently 1, 2, 3, 4, 5, 6, or 7 provided that the sum of p and q is 2, 3, 4, 5, 6, 7, or 8. Alternatively, the bicyclic cycloalkyl group can include a bridged cycloalkyl structure, for example, bicyclo[p.q.r]alkyl where r is 1, 2, or 3 and each of p and q is independently 1, 2, 3, 4, 5, or 6 provided that the sum of p, q, and r is 3, 4, 5, 6, 7, or 8. The cycloalkyl group can be a spiro ring group, for example, spiro[p.q]alkyl where each of p and q is independently 2, 3, 4, 5, 6, or 7 provided that the sum of p and q is 4, 5, 6, 7, 8, or 9. Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, 1-bicyclo[2.2.1.]heptyl, 2-bicyclo[2.2.1.]heptyl, 5-bicyclo[2.2.1.]heptyl, 7-bicyclo[2.2.1.]heptyl, and decalinyl. The cycloalkyl group can be unsubstituted or substituted with one, two, three, four, or five substituents (e.g., optionally substituted cycloalkenyl), and the substituents are independently alkyl; alkenyl; alkynyl; alkoxy; alkylsulfinyl; alkylsulfenyl; alkylsulfonyl; amino; aryl; aryloxy; azide; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heteroalkyl; heterocyclyl; (heterocyclyl)oxy; heteroaryl; hydroxy; nitro; thiol; silyl; cyano; =O; =S; -SO2R (wherein R is optionally substituted amino); =NR' (wherein R' is H, alkyl, aryl, or heterocyclyl); and -CON(R A )2 (wherein each R A is independently H or alkyl, or both Rs A(which together with the atoms to which they are attached form a heterocyclyl). Each of the substituents may itself be unsubstituted or may be substituted with unsubstituted substituent(s) as defined herein for each group.

[0066] As used herein, the term "cycloalkylalkyl" represents an alkyl group substituted with a cycloalkyl group as defined herein. The cycloalkyl moiety and the alkyl moiety may be optionally substituted as described for the individual groups herein.

[0067] As used herein, the term "cycloalkylene" represents a divalent cycloalkyl group. A cycloalkylene optionally substituted is a cycloalkylene optionally substituted as described herein for cycloalkyl.

[0068] As used herein, the term "cycloalkynyl" refers to a monovalent carbocyclic group having one or two carbon-carbon triple bonds and having 8 to 12 carbons, unless otherwise specified. Cycloalkynyl may include one transannular bond or transannular bridge. Non-limiting examples of cycloalkynyl include cyclooctynyl, cyclononynyl, cyclodecynyl, and cyclodecadynyl. The cycloalkynyl group may be unsubstituted or substituted as defined for cycloalkyl (e.g., cycloalkynyl optionally substituted).

[0069] "Disease" or "condition" refers to the state or health condition of a patient or subject that can be treated by the compounds or methods provided herein.

[0070] As used herein, the term "FBXW7" refers to the F-box / WD repeat-containing protein 7 gene, transcript or protein. The FBXW7 mutant gene is also described herein as an FBXW7 gene having an inactivating mutation, which is one that fails to produce a functional FBXW7 protein in cells or produces a reduced amount of the FBXW7 protein.

[0071] As used herein, the term "halo" represents a halogen selected from bromine, chlorine, iodine, and fluorine.

[0072] As used herein, the term "heteroalkyl" refers to an alkyl group, alkenyl group, or alkynyl group that is interrupted once by one or two heteroatoms; interrupted twice, each time independently, by one or two heteroatoms; interrupted three times, each time independently, by one or two heteroatoms; or interrupted four times, each time independently, by one or two heteroatoms. Each heteroatom is independently O, N, or S. In some embodiments, the heteroatom is O or N. No heteroalkyl group contains two adjacent oxygen or sulfur atoms. Heteroalkyl groups may be unsubstituted or substituted (e.g., optionally substituted heteroalkyl). When a heteroalkyl is substituted and the substituent is attached to a heteroatom, the substituent is selected according to the nature and valence of the heteroatom. Thus, a substituent attached to a heteroatom is, within the valence tolerance, =O, -N(R N2 )2, -SO2OR N3 , -SO2R N2 , -SOR N3 , -COOR N3 , an N-protecting group, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclyl, or cyano, where each R N2 is independently H, alkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, or heterocyclyl, and each R N3is independently alkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, or heterocyclyl. Each of these substituents may itself be unsubstituted or may be substituted with unsubstituted substituent(s) as defined herein for each group. When a heteroalkyl is substituted and the substituent is attached to a carbon, the substituent is selected from those described for alkyl, provided that the substituent on the carbon atom attached to the heteroatom is not Cl, Br, or I. The carbon atom is understood to be at the end of the heteroalkyl group.

[0073] As used herein, the term "heteroarylalkyl" represents an alkyl group substituted with a heteroaryl group as defined herein. The heteroaryl moiety and the alkyl moiety may each be optionally substituted as described for the individual groups herein.

[0074] As used herein, the term "heteroarylene" represents a divalent heteroaryl. A heteroarylene that is optionally substituted is a heteroarylene that is optionally substituted as described herein for heteroaryl.

[0075] As used herein, the term "heteroaryloxy" refers to a structure -OR where R is heteroaryl. Heteroaryloxy may be optionally substituted as defined for heterocyclyl.

[0076] As used herein, the term "heterocyclyl", unless otherwise specified, refers to a monocyclic, bicyclic, tricyclic, or tetracyclic ring system having a fused, bridged, and / or spiro 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, or 8-membered ring containing one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. In some embodiments, "heterocyclyl", unless otherwise specified, refers to a monocyclic, bicyclic, tricyclic, or tetracyclic ring system having a fused or bridged 5-membered, 6-membered, 7-membered, or 8-membered ring containing one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur. Heterocyclyl can be aromatic or non-aromatic. Non-aromatic 5-membered heterocyclyl has zero or one double bond, non-aromatic 6- and 7-membered heterocyclyl groups have zero to two double bonds, and non-aromatic 8-membered heterocyclyl groups have zero to two double bonds and / or zero or one carbon-carbon triple bond. Unless otherwise specified, heterocyclyl groups contain 1 to 16 carbon atoms. Certain heterocyclyl groups can contain up to 9 carbon atoms. Examples of non-aromatic heterocyclyl groups include pyrrolinyl, pyrrolidinyl, pyrazolinyl, pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl, homopiperidinyl, piperazinyl, pyridazinyl, oxazolidinyl, isoxazolidinyl, morpholinyl, thiomorpholinyl, thiazolidinyl, isothiazolidinyl, thiazolidinyl, tetrahydrofuranyl, dihydrofuranyl, tetrahydrothienyl, dihydrothienyl, dihydroindolyl, tetrahydroquinolyl, tetrahydroisoquinolyl, pyranyl, dihydropyranyl, dithiazolyl, and the like. When a heterocyclic system has at least one aromatic resonance structure or at least one aromatic tautomer, such a structure is an aromatic heterocyclyl (i.e., heteroaryl).Non-limiting examples of heteroaryl groups include benzimidazolyl, benzofuryl, benzothiazolyl, benzothienyl, benzoxazolyl, furyl, imidazolyl, indolyl, isoindazolyl, isoquinolinyl, isothiazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purinyl, pyrrolyl, pyridinyl, pyrazinyl, pyrimidinyl, quinazolinyl, quinolinyl, thiadiazolyl (e.g., 1,3,4-thiadiazole), thiazolyl, thienyl, triazolyl, tetrazolyl, and the like. The term "heterocyclyl" also refers to heterocyclic compounds having a bridged polycyclic structure in which one or more carbon and / or heteroatoms bridge two non-adjacent members of a monocyclic ring, such as quinuclidine, tropane, or diaza-bicyclo[2.2.2]octane. The term "heterocyclyl" includes bicyclic, tricyclic, and tetracyclic groups in which any of the above heterocycles is fused to one, two, or three carbocyclic rings, such as an aryl ring, cyclohexane ring, cyclohexene ring, cyclopentane ring, cyclopentene ring, or another monocyclic heterocycle. Examples of fused heterocyclyls include 1,2,3,5,8,8a-hexahydroindolizine; 2,3-dihydrobenzofuran; 2,3-dihydroindole; and 2,3-dihydrobenzothiophene. The heterocyclyl group may be unsubstituted or substituted with one, two, three, four, or five substituents independently selected from the group consisting of alkyl; alkenyl; alkynyl; alkoxy; alkylsulfinyl; alkylsulfenyl; alkylsulfonyl; amino; aryl; aryloxy; azide; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heteroalkyl; heterocyclyl; (heterocyclyl)oxy; hydroxy; nitro; thiol; silyl; cyano; -C(O)R or -SO2R (wherein R is amino or alkyl); =O; =S; =NR' (wherein R' is H, alkyl, aryl, or heterocyclyl). Each of the substituents may itself be unsubstituted or may be substituted with one or more unsubstituted substituents as defined herein for each group.

[0077] As used herein, the term "heterocyclylalkyl" represents an alkyl group substituted with a heterocyclyl group as defined herein. The heterocyclyl moiety and the alkyl moiety may each be optionally substituted as described for the individual groups herein.

[0078] As used herein, the term "heterocyclylene" represents a divalent heterocyclyl. A heterocyclylene that is optionally substituted is a heterocyclylene that is optionally substituted as described herein for heterocyclyl.

[0079] As used herein, the term "(heterocyclyl)oxy", unless otherwise specified, represents a chemical substituent of the formula -OR, wherein R is a heterocyclyl group. (Heterocyclyl)oxy may be optionally substituted as defined for heterocyclyl.

[0080] As used herein synonymously, the terms "hydroxyl" and "hydroxy" represent an -OH group.

[0081] As used herein, the term "isotope enrichment" refers to a pharmaceutically active agent in which the isotope content of one isotope at a given position within a molecule is at least 100 times greater than the natural abundance of this isotope. For example, a composition that is isotope enriched for deuterium includes an active agent having deuterium at the position of at least one hydrogen atom in an amount that is at least 100 times greater than the natural abundance of deuterium. Preferably, the isotope enrichment of deuterium is such that the deuterium is at least 1000 times greater than the natural abundance. More preferably, the isotope enrichment of deuterium is such that the deuterium is at least 4000 times (e.g., at least 4750 times, e.g., up to 5000 times) greater than the natural abundance.

[0082] As used herein, the term "leukemia" refers broadly to a progressive malignant disease of the hematopoietic organs, generally characterized by the abnormal proliferation and development of white blood cells and their precursors in the blood and bone marrow. Leukemia is generally clinically classified based on (1) the duration and characteristics of the disease (acute or chronic); (2) the cell type involved; bone marrow (myeloid), lymph (lymphoid), or monocyte; and (3) the presence or absence of an increase in the number of abnormal cells, leukemic or non-leukemic (sub-leukemic).

[0083] As used herein, the term "lymphoma" refers to cancers arising from cells of immune origin.

[0084] As used herein, the term "melanoma" is interpreted to mean tumors arising from the melanocyte system of the skin and other organs.

[0085] As used herein, the term "Myt1" refers to the membrane-bound tyrosine and threonine-specific cdc2 inhibitory kinase (Myt1) (gene name PKMYT1).

[0086] As used herein, the term "Myt1 inhibitor" refers to a compound that, whether in vitro, in cell culture, or in an animal, when contacted with the enzyme Myt1, results in a decrease in the activity of Myt1 such that the measured Myt1 IC 50 is 10 μM or less (e.g., 5 μM or less, or 1 μM or less). In certain Myt1 inhibitors, the Myt1 IC 50 can be 100 nM or less (e.g., 10 nM or less, or 3 nM or less), and in some cases can be as low as 100 pM or 10 pM. Preferably, the Myt1 IC 50 is from 1 nM to 1 μM (e.g., 1 - 750 nM, 1 - 500 nM, or 1 - 250 nM). Even more preferably, the Myt1 IC 50 is less than 20 nM (e.g., 1 - 20 nM).

[0087] As used herein, the term "nitro" represents the -NO2 group.

[0088] As used herein, the term "oxo" represents a divalent oxygen atom (e.g., the structure of oxo may be shown as =O).

[0089] As used herein, the term "Ph" represents phenyl.

[0090] As used herein, the term "pharmaceutical composition" refers to a composition that contains a compound described herein, is formulated with pharmaceutically acceptable excipients, and is manufactured or sold with the approval of a government regulatory authority as part of a therapeutic regimen for the treatment of diseases in mammals. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage forms (e.g., tablets, capsules, caplets, gelcaps, or syrups); for topical administration (e.g., as creams, gels, lotions, or ointments); for intravenous administration (e.g., as a sterile solution free of particulate embolizing substances and in a solvent system suitable for intravenous use); or in any other suitable formulation described herein.

[0091] As used interchangeably herein, the terms "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refer to any component other than the compounds described herein that has the properties of being non-toxic and non-inflammatory to a patient (e.g., a vehicle in which an active compound can be suspended or dissolved). Excipients can include, for example, anti-adhesion agents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (pigments), emollients, emulsifiers, diluents, film-forming agents or coatings, flavoring agents, fragrances, glidants (flow activators), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, or water of hydration. Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, dibasic calcium phosphate, calcium stearate, croscarmellose, crospovidone, cysteine, ethyl cellulose, gelatin, hydroxypropyl cellulose, hydroxypropyl methylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methyl cellulose, methyl paraben, microcrystalline cellulose, polyethylene glycol, polyvinyl pyrrolidone, povidone, pregelatinized starch, propyl paraben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethyl cellulose, sodium citrate, sodium starch glycolate, sorbitol, corn starch, stearic acid, sucrose, talc, titanium dioxide, vitamin A, vitamin E, vitamin C, and xylitol.

[0092] As used herein, the term "pharmaceutically acceptable salt" means that the salt is suitable for use in contact with human and animal tissues within the scope of sound medical judgment, without undue toxicity, irritation, allergic response, etc., and represents a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, pharmaceutically acceptable salts are described in Berge et al., J. Pharmaceutical Sciences 66:1-19, 1977 and Pharmaceutical Salts: Properties, Selection, and Use, (Eds. P. H. Stahl and C. G. Wermuth), Wiley-VCH, 2008. The salts can be prepared in situ during the final isolation and purification of the compounds described herein or separately by reacting the free base with a suitable organic acid. Representative acid addition salts include acetate, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, fumarate, glucoheptonate, glycerophosphate, hemisulfate, heptonate, hexanoate, hydrobromide, hydrochloride, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, toluenesulfonate, undecanoate, valerate, etc.Representative alkali metal salts and alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, etc., as well as non-toxic ammonium, quaternary ammonium, and amine cations, such as, but not limited to, ammonium, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, triethylamine, ethylamine, and the like.

[0093] As used herein, the terms "premalignant" or "precancerous" refer to conditions that are not malignant but have a tendency to become malignant.

[0094] As used herein, the term "protecting group" refers to a group that is intended to prevent a hydroxy, amino, or carbonyl group from participating in one or more unwanted reactions during chemical synthesis. As used herein, the term "O - protecting group" refers to a group that is intended to prevent a hydroxy group or a carbonyl group from participating in one or more unwanted reactions during chemical synthesis. As used herein, the term "N - protecting group" refers to a group that is intended to prevent a nitrogen - containing (e.g., amino, amide, N - H heterocycle, or hydrazine) group from participating in one or more unwanted reactions during chemical synthesis. Commonly used O - protecting groups and N - protecting groups are described in Greene, "Protective Groups in Organic Synthesis," 3rd Edition (John Wiley & Sons, New York, 1999), which is hereby incorporated by reference. Exemplary O - protecting groups and N - protecting groups include alkanoyl groups, aroyl groups, or carbamyl groups, such as formyl, acetyl, propionyl, pivaloyl, t - butylacetyl, 2 - chloroacetyl, 2 - bromoacetyl, trifluoroacetyl, trichloroacetyl, phthalyl, o - nitrophenoxyacetyl, α - chlorobutyryl, benzoyl, 4 - chlorobenzoyl, 4 - bromobenzoyl, t - butyldimethylsilyl, tri - iso - propylsilyloxymethyl, 4,4’ - dimethoxytrityl, isobutyryl, phenoxyacetyl, 4 - isopropylphenoxyacetyl, dimethylformamidine, and 4 - nitrobenzoyl.

[0095] Exemplary O - protecting groups for protecting carbonyl - containing groups include, but are not limited to, acetals, acylals, 1,3 - dithians, 1,3 - dioxanes, 1,3 - dioxolanes, and 1,3 - dithiolanes.

[0096] As other O-protecting groups, there may be mentioned, but are not limited to, substituted alkyl, aryl, and aryl-alkyl ethers (e.g., trityl; methylthiomethyl; methoxymethyl; benzyloxymethyl; siloxymethyl; 2,2,2-trichloroethoxymethyl; tetrahydropyranyl; tetrahydrofuranyl; ethoxyethyl; 1-[2-(trimethylsilyl)ethoxy]ethyl; 2-trimethylsilylethyl; t-butyl ether; p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, benzyl, p-methoxybenzyl, and nitrobenzyl); silyl ethers (e.g., trimethylsilyl; triethylsilyl; triisopropylsilyl; dimethylisopropylsilyl; t-butyldimethylsilyl; t-butyldiphenylsilyl; tribenzylsilyl; triphenylsilyl; and diphenylmethylsilyl); carbonates (e.g., methyl, methoxymethyl, 9-fluorenylmethyl; ethyl; 2,2,2-trichloroethyl; 2-(trimethylsilyl)ethyl; vinyl, allyl, nitrophenyl; benzyl; methoxybenzyl; 3,4-dimethoxybenzyl; and nitrobenzyl).

[0097] As other N-protecting groups, there are asymmetric auxiliaries such as protected or unprotected D-amino acids, L-amino acids, or D,L-amino acids such as alanine, leucine, phenylalanine; sulfonyl-containing groups such as benzenesulfonyl, p-toluenesulfonyl, etc.; carbamate-forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenylyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl, t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl, etc.; aryl-alkyl groups such as benzyl, p-methoxybenzyl, 2,4-dimethoxybenzyl, triphenylmethyl, benzyloxymethyl, etc.; silylalkyl acetal groups such as [2-(trimethylsilyl)ethoxy]methyl; and silyl groups such as trimethylsilyl, etc., but are not limited thereto. Useful N-protecting groups are formyl, acetyl, benzoyl, pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl, dimethoxybenzyl, [2-(trimethylsilyl)ethoxy]methyl (SEM), tetrahydropyranyl (THP), t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

[0098] The term "tautomer" often refers to structural isomers that are readily interconverted by the migration of a proton. Tautomers are distinct chemical species that can be identified by differences in spectroscopic properties but usually cannot be separated individually. Non-limiting examples of tautomers include keto-enol, enamine-imine, amide-imino acid, nitroso-oxime, ketene-enol, and amino acid-carboxylic acid ammonium.

[0099] Generally, the term "sarcoma" refers to a tumor composed of substances such as embryonic connective tissue and usually consisting of dense, solid cells embedded in a fibrous or homogeneous substance.

[0100] As used herein, the term "subject" refers to a human or non-human animal (e.g., a mammal) that has been determined by a qualified professional (e.g., a physician or nurse), regardless of the presence or absence of clinical examination(s) of a subject-derived sample(s) known in the art, to be suffering from or at risk of a disease or condition. Preferably, the subject is human. Non-limiting examples of diseases and conditions include diseases having symptoms of excessive cell proliferation, such as cancer.

[0101] As used herein, "treatment" or "treating" refers to the medical management of a subject with the intention of improving, ameliorating, stabilizing, preventing, or curing a disease or condition. This term encompasses active treatment (treatment directed at improving a disease or condition); causal treatment (treatment directed at the cause of a related disease or condition); symptomatic treatment (treatment designed to relieve the symptoms of a disease or condition); prophylactic treatment (treatment directed at minimizing or suppressing in part or completely the onset of a related disease or condition); and adjuvant treatment (treatment used to supplement another therapy).

Brief Description of the Drawings

[0102]

Figure 1A

Figure 1B

Figure 2A

Figure 2B

Figure 3A

Figure 3B

Figure 3C

Figure 4A

Figure 4B

Figure 4C

Figure 5

Figure 6

Figure 7

[0103] Generally, the present invention provides compounds, pharmaceutical compositions containing them, methods for preparing the compounds, and methods of use. The compounds of the present invention can be Myt1 inhibitors. Using such compounds, Myt1 can be inhibited in cells, such as cells of a subject (e.g., cells overexpressing CCNE1 or having inactivating mutations in the FBXW7 gene). The subject can be one in need of treatment for a disease or disorder, such as a disease or disorder having symptoms of cell overproliferation, such as cancer. The Myt1 inhibitory activity of the compounds disclosed herein is useful for the treatment of subjects in need of cancer treatment.

[0104] Myt1 is a cell cycle regulatory kinase that is mainly localized in the endoplasmic reticulum and Golgi complex. It is part of the Wee family of kinases, including Wee1 and Wee1b. It is involved in the negative regulation of the CDK1-cyclin B complex, which promotes the progression of cells from the G2 phase to the mitotic (M) phase of the cell cycle. During DNA damage, Myt1, together with Wee1 (which mediates phosphorylation only at Tyr15), promotes phosphorylation of CDK1 (both Tyr15 and Thr14 of CDK1) as part of the G2 checkpoint response, which maintains the kinase complex in an inactive state in the G2 phase and prevents progression to mitosis until the damage is repaired. In addition, it has been proposed that Myt1 directly interacts with the CDK1 complex in the cytoplasm to prevent their nuclear translocation, resulting in inhibition of cell cycle progression.

[0105] Myt1 is essential in many cancer cells and has therefore been implicated as a potentially important cancer target. Overexpression of Myt1 has been observed in various cancers, including hepatocellular carcinoma and clear cell renal cell carcinoma. Myt1 downregulation plays a minor role in normal cells but plays a more prominent role in cells exposed to DNA damage. In addition, cells that exhibit high levels of replication stress in addition to G1 checkpoint dysregulation may be particularly sensitive to loss of Myt1 function, as these cells would be prone to premature progression into mitosis with compromised genomic material, leading to mitotic cell death.

[0106] Inhibitors of Myt1, a regulator of the G2-M transition, may be particularly useful in treating tumors harboring CCNE1 amplification or FBXW7 loss-of-function mutations using synthetic lethal therapeutic strategies.

[0107] Cyclin E1 (encoded by the CCNE1 gene) is involved in the cell cycle transition from G1 to S phase. During the latter half of the G1 phase of the cell cycle, it complexes with cyclin-dependent kinase 2 (CDK2) to promote E2F transcription factor activation and progression into S phase. During the normal cell cycle, cyclin E1 levels are tightly regulated, accumulating at the G1 / S phase transition and being completely degraded by the end of S phase. Cell cycle-dependent proteasomal degradation of cyclin E1 is mediated by SCF FBW7 This is mediated by the ubiquitin ligase complex. Upon activation in late G1, the cyclin E1 / CDK2 complex promotes entry into S phase by phosphorylating and inactivating RB1 and subsequently releasing E2F transcription factors. S phase is promoted by E2F-mediated transcription of numerous genes involved in DNA replication, including the pre-replication complex subunits ORC1, CDC6, CDT1, and MCM helicase factors.

[0108] CCNE1 is frequently amplified and / or overexpressed in human cancers (Figure 1). CCNE1 amplification has been reported in several cancer types including endometrial cancer, ovarian cancer, breast cancer, and gastric cancer, with frequencies ranging from 5% to 40%. Importantly, in these disease states, cyclin E1 has been confirmed in numerous studies to be a promoter of tumorigenesis, and CCNE1 amplification is observed in more aggressive subtypes including uterine carcinosarcoma (UCS, ~40%), uterine serous carcinoma (USC, ~25%), high-grade serous ovarian carcinoma (HGSOC, ~25%), and triple-negative breast cancer (TNBC, ~8%). Patients with evidence of cyclin E1 overexpression in tumor biopsies by immunohistochemistry and / or genomic copy number analysis have lower overall survival compared to patients with normal cyclin E1 levels. HGSOC patients with cyclin E1 overexpression have a low response rate to cisplatin, the current standard treatment.

[0109] SCF regulated by the cell cycle FBW7 Defective proteolysis of cyclin E1 by the ubiquitin ligase complex is another mechanism of CCNE1 overexpression observed in tumors. The F-box protein gene FBXW7 is frequently mutated in several types of cancer including endometrial cancer, colorectal cancer, and gastric cancer, with frequencies ranging from 5% to 35% (Figure 2). Similar to CCNE1, mutations promoting FBXW7 are observed in more aggressive subtypes of endometrial cancer including UCS (~35%) and USC (~25%). In cancer, FBXW7 has diverse loss-of-function mutations including truncating mutations scattered across the gene and missense mutations within the cyclin E1 recognition WD40 repeat. FBW7 functions as a homodimer in the SCF complex, and many of the deleterious missense mutations within the WD40 repeat are mostly heterozygous and dominant negative. Unexpectedly, several recurrent hotspot missense mutations have been found in the WD40 repeat, including R465, R479, and R505, all of which interfere with cyclin E1 binding and ubiquitination.

[0110] Overexpression of cyclin E1 and / or loss of FBXW7 function are thought to promote tumorigenesis by inducing genomic instability (e.g., increased origin firing, poor nucleotide pools, transcription-replication conflicts, and / or fork instability). Overexpression of cyclin E1 has been shown to induce replication stress characterized by deceleration or stalling of replication forks and loss of heterozygosity at fragile sites. The main mechanisms by which cyclin E1 overexpression causes replication stress are increased origin firing in the early S phase and subsequent depletion of replication factors including nucleotide pools. The overall decrease in replication proteins and nucleotides reduces fork progression, leading to stalling and subsequent collapse or regression.

[0111] The compounds of the present invention are, for example, compounds of formula (I):

Chemical formula

[0112] Preferably, the compound of formula (I) is enriched for the atropisomer of formula (IA):

Chemical formula

[0113] The compounds of the present invention are, for example, compounds of formula (II): [ka] wherein all variables are as defined herein. It could be.

[0114] Preferably, the compound of formula (II) is an atropisomer of formula (IIA): [ka] wherein all variables are as defined herein. It is enriched for

[0115] The compounds of the present invention are, for example, compounds of formula (III): [ka] [During the ceremony, R 2A is hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkenyl, optionally substituted C 2-6 Alkynyl, optionally substituted C 3-8 Cycloalkyl, optionally substituted C 3-8 Cycloalkenyl, optionally substituted C 2-9 Heterocyclyl, optionally substituted C 2-9 Heterocyclyl C 1-6 Alkyl, optionally substituted C 6-10 Aryl, optionally substituted C 1-9 Heteroaryl, optionally substituted C 1-9 Heteroaryl C 1-6 Alkyl, halogen, -N(R 7 )2, -OR 7 , -C(O)N(R 8 )2, -SO2N(R8 )2, -SO2R 7A , or -QR 7B is] It could be.

[0116] Preferably, the compound of formula (III) is an atropisomer of formula (IIIA): [ka] It is enriched for

[0117] The compound of the invention can be, for example, a compound listed in Table 1 below, or a pharmaceutically acceptable salt thereof. [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8] [Table 1-9] [Table 1-10]

Table 1-11

Table 1-12

Table 1-13

Table 1-14

Table 1-15

Table 1-16

Table 1-17

Table 1-18

Table 1-19

Table 1-20

Table 1-21

Table 1-22

Table 1-23

Table 1-24

Table 1-25

Table 1-26

Table 1-27

Table 1-28

Table 1-29

[0118] The present invention includes, where possible, the individual diastereomers, enantiomers, epimers, and atropisomers of the compounds disclosed herein, as well as mixtures of its diastereomers and / or enantiomers including racemic mixtures. Although specific stereochemistry is disclosed herein as preferred, other stereoisomers including diastereomers, enantiomers, epimers, atropisomers, and mixtures thereof may also have utility in the treatment of Myt1-mediated diseases. Inactive or less active diastereoisomers and enantiomers may be useful, for example, in scientific studies related to receptors and activation mechanisms.

[0119] It is understood that certain molecules may exist in multiple tautomeric forms. The present invention includes all tautomers even if only one tautomer is shown in the examples.

[0120] The present invention also includes pharmaceutically acceptable salts of the compounds, as well as pharmaceutical compositions containing the compounds and pharmaceutically acceptable carriers. Such compounds are particularly useful, for example, in certain types of cancer and for delaying the progression of cancer after it has developed in a patient.

[0121] The compounds disclosed herein can be used in pharmaceutical compositions containing (a) the compound(s) or a pharmaceutically acceptable salt thereof, and (b) a pharmaceutically acceptable carrier. The compounds can be used in pharmaceutical compositions containing one or more other active pharmaceutical ingredients. The compounds can be used in pharmaceutical compositions in which the compound disclosed herein or a pharmaceutically acceptable salt thereof is the only active ingredient.

[0122] Optical isomers - Diastereomers - Geometric isomers - Tautomers The compounds disclosed herein may, for example, contain one or more stereocenters and may exist as racemates, racemic mixtures, single enantiomers, individual diastereomers, and mixtures of diastereomers and / or enantiomers. The present invention includes all such isomeric forms of the compounds disclosed herein. All possible stereoisomers in mixtures, as well as those possible as pure compounds or partially purified compounds (e.g., enantiomers and / or diastereomers), are intended to be included within the scope of the present invention (i.e., as pure compounds or all possible combinations of stereocenters in mixtures).

[0123] Some of the compounds described herein may contain bonds with rotational hindrance, which can be advantageous in that two individual rotational isomers, i.e., atropisomers, can be separated and detected as having different biological activities. All possible atropisomers are intended to be encompassed within the scope of the present invention.

[0124] Some of the compounds described herein may contain olefinic double bonds and are intended to include both E and Z geometric isomers unless otherwise specified.

[0125] Some of the compounds described herein may exist with different points of attachment of hydrogen, referred to as tautomers. One example is a ketone and its enol form, known as keto-enol tautomers. The individual tautomers as well as mixtures thereof are encompassed by the present invention.

[0126] Compounds disclosed herein that possess one or more asymmetric centers can be separated into diastereoisomers, enantiomers, etc. by methods well known in the art.

[0127] Alternatively, enantiomers and other compounds containing chiral centers may be synthesized by stereospecific synthesis using optically pure starting materials and / or reagents of known configuration.

[0128] Metabolites-prodrugs The present invention includes therapeutically active metabolites, which are themselves encompassed within the scope of the claims. The present invention also includes prodrugs, which are compounds that are converted to the claimed compounds during or after administration to a patient. The claimed chemical structures of this use may themselves be prodrugs in some cases.

[0129] Isotopically enriched derivatives The present invention includes molecules that are isotopically enriched at one or more positions within the molecule, and therefore, compounds enriched with deuterium are encompassed within the scope of the claims.

[0130] Methods for preparing the compounds of the present invention The compounds of the present invention can be prepared using reactions and techniques known in the art and those described herein. Those skilled in the art will recognize that the methods for preparing the compounds of the present invention described herein are not limiting and that steps within the methods may be interchangeable without affecting the structure of the final product.

[0131] Method A The compounds of the present invention can be prepared as shown in Scheme A and described herein. The amino group of commercially available 5-bromo-6-chloropyrazine-2-amine can be converted to a hydroxyl, which can be benzylated with benzyl bromide in the presence of base to produce key intermediate B. The bromo can be displaced with an aromatic amine under metal-mediated conditions. Depending on the nature of the aryl amine, placement of a protecting group may be required prior to this reaction. The chloro can be displaced with malononitrile under metal-mediated conditions to produce aminopyrrole intermediate C. The nitrile can be hydrolyzed to a carboxamide by treatment with acid, with concomitant cleavage of the benzyl group. The resulting hydroxyl can be converted to a triflate to produce triflate key intermediate D, which can be derivatized in several different ways to yield compounds of the present invention. For example, metal-mediated coupling or S N Ar substitution with R 1 A group may be introduced into R 1 Depending on the nature of the group, it may be necessary to place a protecting group prior to the triflate derivatization reaction. 1 If the group contains unsaturation, a hydrogenation reaction may be required to obtain the compounds of the present invention. 1 If the group has a protecting group, deprotection step(s) using acid, base, and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, atropisomeric mixtures may be obtained. In such cases, isolation of the atropisomer of interest may be required to obtain the compounds of the present invention. Alternatively, an atropisomerically pure intermediate may be isolated, which may be further derivatized to obtain the compounds of the present invention. For example, key intermediate D may be purified by chiral chromatography to obtain an atropisomerically pure intermediate, which may be manipulated similarly to intermediate D above to obtain the compounds of the present invention. [ka]

[0132] Method B The compounds of the present invention are shown in Scheme B and can be prepared as described herein. The chloro of Intermediate B can be substituted with an aromatic amine under S N Ar conditions. Depending on the nature of the arylamine, placement of a protecting group may be required prior to this reaction. The bromo can be substituted with malononitrile under metal-mediated conditions to produce an aminopyrrole. OBn can be hydrogenolyzed to produce a major Intermediate E that can be derivatized in a plurality of different ways to obtain the compounds of the present invention after nitrile hydrolysis. For example, a group can be introduced to R 2 using Mitsunobu or alkylation conditions. Alternatively, Intermediate E can be converted to a triflate to produce a triflate major Intermediate F that can be derivatized in a plurality of different ways to obtain the compounds of the present invention after nitrile hydrolysis. For example, a group can be introduced to R 2 using metal-mediated coupling. Depending on the nature of the R 2 group, placement of a protecting group may be required prior to the hydroxyl or triflate derivatization reaction. If the R 2 group has unsaturation, a hydrogenation reaction may be required to obtain the compounds of the present invention. If the arylamine and / or the R 2 group has a protecting group, a deprotection step(s) using an acid, base, and / or fluoride may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, a mixture of atropisomers may be obtained. In such cases, it may be necessary to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention.

Chemical formula

[0133] Method C The compounds of the present invention are shown in Scheme C and can be prepared as described herein. The 2-chloro of commercially available 3-bromo-2,6-dichloropyridine can be S NIt can be substituted with malononitrile under Ar conditions. Bromo can be substituted with an aromatic amine under metal-mediated conditions to produce an aminoazaindole. Depending on the nature of the arylamine, placement of a protecting group may be required prior to this reaction. The protecting group(s) can be removed before the nitrile can be hydrolyzed to a carboxamide by treatment with acid to obtain intermediate G. The remaining chloro can be derivatized in a plurality of different ways to yield the compounds of the present invention. For example, a group based on R 1 can be introduced using metal-mediated coupling. Depending on the nature of the R 1 group, placement of a protecting group may be required prior to the chloro-derivatization reaction. When the R 1 group has unsaturation, a hydrogenation reaction may be required to obtain the compounds of the present invention. When the arylamine and / or the R 1 group has a protecting group, a deprotection step(s) using an acid, a base and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, a mixture of atropisomers may be obtained. In such cases, it may be necessary to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention.

Chemical formula

[0134] Method D The compounds of the present invention are shown in Scheme D and can be prepared as described herein. The halogen at the 2-position of a suitably substituted 5-nitropyridine is S NIt can be displaced with an aromatic amine under Ar or metal-mediated C-N coupling conditions. Depending on the nature of the aryl amine, placement of a protecting group may be required prior to this reaction. The 3-bromo can be displaced with malononitrile under palladium-mediated conditions to generate an aminoazaindole. The resulting amino group can be protected with a suitable protecting group, e.g., BOC. The nitro can be reduced, and the resulting amino can be converted to a halogen under Sandmeyer conditions to give a halogenated derivative. The N-protecting group of the aminopyrrole can be cleaved, and the nitrile can be hydrolyzed to a carboxamide to give intermediate I, which can be derivatized in several different ways to yield compounds of the invention. For example, metal-mediated coupling can be used to form R 1 A group may be introduced into R 1 If the group contains unsaturation, a hydrogenation reaction may be required to obtain the compounds of the invention. 1 Depending on the nature of the groups, placement of protecting groups may be required prior to the halogen derivatization reaction. 1 If the group has a protecting group, deprotection step(s) using acid, base and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, atropisomeric mixtures may be obtained. In such cases, isolation of the atropisomer of interest may be required to obtain the compounds of the present invention. Alternatively, atropisomerically pure intermediates may be isolated, which may be further derivatized to obtain the compounds of the present invention. [ka]

[0135] Method E Compounds of the invention can be prepared as shown in Scheme E and described herein. One chloro of commercially available 2,3-dichloro-pyrazine is S N The remaining chloro can be replaced by malononitrile under Ar or palladium-mediated conditions. NUnder Ar conditions or palladium-mediated conditions, it can be substituted with an aromatic amine to produce aminopyrrole. Depending on the nature of the arylamine, placement of a protecting group may be required prior to this reaction. Hydrolysis of the nitrile can be carried out under acidic or basic conditions to obtain the compounds of the present invention. If the arylamine has a protecting group, a deprotection step(s) using an acid, base and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, a mixture of atropisomers may be obtained. In such cases, it may be necessary to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention.

Chemical formula

[0136] Method F The compounds of the present invention are shown in Scheme F and can be prepared as described herein. One chloro of commercially available 2,3-dichloropyrazine can be substituted with malononitrile under Ar N conditions or palladium-mediated conditions. The other chloro can be substituted with an aromatic amine under Ar N conditions or palladium-mediated conditions to produce aminopyrrole. Depending on the nature of the arylamine, placement of a protecting group may be required prior to this reaction. The pyrazine ring can be brominated using a suitable brominating reagent such as NBS. Hydrolysis of the nitrile can be carried out under acidic or basic conditions, and the protecting group is cleaved to obtain a major intermediate H that can be derivatized in a plurality of different ways to obtain the compounds of the present invention. For example, an R 2 group can be introduced using a metal-mediated coupling. Depending on the nature of the R 2 group, placement of a protecting group may be required prior to the bromo-derivatization reaction. If the R 2 group has unsaturation, a hydrogenation reaction may be required to obtain the compounds of the present invention. The arylamine and / or R 2If the group has a protecting group, deprotection step(s) using acid, base and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, atropisomeric mixtures may be obtained. In such cases, isolation of the atropisomer of interest may be required to obtain the compounds of the present invention. Alternatively, atropisomerically pure intermediates may be isolated, which may be further derivatized to obtain the compounds of the present invention. [ka]

[0137] Method G Compounds of the invention can be prepared as shown in Scheme G and described herein. The chloro in commercially available 2-chloro-3-bromopyridine is S N It can be replaced by malononitrile under Ar conditions. N The arylamine may be substituted with an aromatic amine under Ar or palladium-mediated conditions to produce an aminopyrrole. Depending on the nature of the arylamine, the placement of a protecting group may be required prior to this reaction. Hydrolysis of the nitrile may be carried out under acidic or basic conditions to give the compounds of the present invention. In the case of arylamine groups bearing protecting groups, deprotection step(s) using acid, base, and / or fluorine may be required to give the compounds of the present invention. Depending on the nature of the arylamine, a mixture of atropisomers may be obtained. In such cases, isolation of the atropisomer of interest may be required to give the compounds of the present invention. Alternatively, atropisomerically pure intermediates may be isolated, which may be further derivatized to give the compounds of the present invention. [ka]

[0138] Method H The compounds of the present invention are shown in Scheme H and can be prepared as described herein. The major intermediate C can be brominated using a suitable brominating reagent such as NBS. The nitrile can be hydrolyzed to the carboxamide with concomitant cleavage of the benzyl group by treatment with an acid. The resulting hydroxyl can be converted to the triflate. The bromo and triflate can be derivatized sequentially with different groups or the bromo and triflate can be derivatized simultaneously with the same group. The bromo and triflate can be derivatized in a plurality of different ways to obtain the compounds of the present invention. For example, using metal-mediated coupling, R 1 and / or R 2 groups may be introduced sequentially or simultaneously. Depending on the nature of the R 1 and / or R 2 groups, placement of a protecting group may be required prior to the bromo or triflate derivatization reaction. If the arylamine, R 1 and / or R 2 groups have a protecting group, a deprotection step(s) using an acid, base and / or fluoride may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine used to prepare intermediate C, a mixture of atropisomers may be obtained. In such cases, it may be necessary to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropisomerically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention.

Chemical formula

[0139] Method I The compounds of the present invention are shown in Scheme I and can be prepared as described herein. The chloro of commercially available 3-bromo-2-chloro-5-(trifluoromethyl)pyridine is S NThe arylamine may be substituted with an aromatic amine under Ar or palladium-mediated conditions. Depending on the nature of the arylamine, the placement of a protecting group may be required prior to this reaction. Bromo may be substituted with malononitrile under palladium-mediated conditions to generate an aminopyrrole. Hydrolysis of the nitrile may be carried out under acidic or basic conditions to give the compounds of the present invention. If the arylamine group has a protecting group, deprotection step(s) using acid, base, and / or fluorine may be required to give the compounds of the present invention. Depending on the nature of the arylamine, a mixture of atropisomers may be obtained. In such cases, isolation of the atropisomer of interest may be required to give the compounds of the present invention. Alternatively, an atropisomerically pure intermediate may be isolated, which may be further derivatized to give the compounds of the present invention. [ka]

[0140] Method J Compounds of the invention are shown in Scheme J and can be prepared as described herein. Key intermediate C can be halogenated using a suitable halogenating reagent such as NBS or NIS. The halogen can be derivatized in several different ways. For example, metal-mediated coupling can be used to derivatize R 2 A group may be introduced into R. The nitrile can be hydrolyzed to a carboxamide by treatment with acid, with concomitant cleavage of the benzyl group. The resulting hydroxyl can be converted to a triflate. The triflate can be derivatized in several different ways to give the compounds of the invention. For example, metal-mediated coupling can be used to introduce R 1 A group may be introduced into R 1 and / or R 2 Depending on the nature of the groups, placement of protecting groups may be required prior to the halogen and / or triflate derivatization reaction. 1 and / or R 2When the base has a protecting group, a deprotection step(s) using an acid, a base and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine used in the preparation of Intermediate C, a mixture of atropisomers may be obtained. In such cases, it may be necessary to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention.

Chemical formula

[0141] Method K The compounds of the present invention are shown in Scheme K and can be prepared as described herein. The fluorine of 3-bromo-2-fluoro-pyridine can be substituted with an aromatic amine under S N Ar conditions. Depending on the nature of the arylamine, placement of a protecting group may be required prior to this reaction. The bromo can be substituted with malononitrile under palladium-mediated conditions to produce an aminoazaindole. Hydrolysis of the nitrile can be carried out under acidic or basic conditions to obtain the compounds of the present invention. The arylamine or R 1 When the group has a protecting group, a deprotection step(s) using an acid, a base and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, a mixture of atropisomers may be obtained. In such cases, it may be necessary to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention.

Chemical formula

[0142] Method L The compounds of the present invention are shown in Scheme L and can be prepared as described herein. The triflate of the major intermediate D can be derivatized in a plurality of different ways to obtain the compounds of the present invention. For example, a group can be introduced for R 1 using a metal-mediated coupling. Pyrazine can be brominated using a suitable brominating reagent such as NBS. Bromo can be derivatized in a plurality of different ways to obtain the compounds of the present invention. For example, a group can be introduced for R 2 using a metal-mediated coupling. Depending on the nature of the R 1 and / or R 2 groups, placement of a protecting group may be required prior to the triflate and / or bromo derivatization reaction. When the arylamine, R 1 and / or R 2 groups have a protecting group, a deprotection step(s) using an acid, base and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine used in the preparation of intermediate D, a mixture of atropisomers may be obtained. In such cases, it may be necessary to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention.

Chemical formula

[0143] Method M The compounds of the present invention are shown in Scheme M and can be prepared as described herein. The nitrile of the major intermediate C can afford a ketone by treatment with a Grignard reagent. The benzyl group can be cleaved under acidic conditions. The resulting hydroxyl can be converted to a triflate to produce a triflate that can be derivatized in a plurality of different ways to afford the compounds of the present invention. For example, a group can be introduced for R 1 using a metal-mediated coupling. Depending on the nature of the R 1 group, placement of a protecting group may be required prior to the triflate derivatization reaction. R 1If the group contains unsaturation, a hydrogenation reaction may be required to obtain the compounds of the present invention. 1 If the group bears a protecting group, deprotection step(s) using acid, base, and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine used to prepare intermediate C, atropisomeric mixtures may be obtained. In such cases, isolation of the atropisomer of interest may be necessary to obtain the compounds of the present invention. Alternatively, atropisomerically pure intermediates may be isolated, which may be further derivatized to obtain the compounds of the present invention. [ka]

[0144] Method N The compounds of the present invention can be prepared as shown in Scheme N and described herein. The amino of the aminopyrroles described herein can be replaced with a proton under diazotization conditions. The nitrile can be hydrolyzed to a carboxamide under acidic or basic conditions to provide the compounds of the present invention. The arylamine, R 1 and / or R 2 If the group has a protecting group, deprotection step(s) using acid, base, and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the N-aryl group, atropisomeric mixtures may be obtained. In such cases, isolation of the atropisomer of interest may be required to obtain the compounds of the present invention. Alternatively, atropisomerically pure intermediates may be isolated, which may be further derivatized to obtain the compounds of the present invention. [ka]

[0145] Method O The compounds of the present invention are shown in Scheme O and can be prepared as described herein. 2-Aminopyridine can be converted to 2-hydroxypyridine, which can be converted to 2-bromopyridine. 2-Bromo can be substituted with an aromatic amine under palladium-mediated conditions. Depending on the nature of the arylamine, placement of a protecting group may be required prior to this reaction. 3-Bromo can be substituted with malononitrile under palladium-mediated conditions to produce aminopyrrole. Hydrolysis of the nitrile can be carried out under acidic or basic conditions to obtain the compounds of the present invention. If the arylamine group has a protecting group, a deprotection step(s) using an acid, base, and / or fluorine may be required to obtain the compounds of the present invention. Depending on the nature of the arylamine, a mixture of atropisomers may be obtained. In such cases, chiral chromatography may be carried out to isolate the atropisomer of interest to obtain the compounds of the present invention. Alternatively, an atropically pure intermediate may be isolated and further derivatized to obtain the compounds of the present invention. [Chemical formula]

[0146] Therapeutic methods The compounds of the present invention can be used for the treatment of diseases or conditions that depend on the activity of Myt1 (gene name PKMYT1) (for example, cancers in which CCNE1 is overexpressed or has an inactivating mutation in the FBXW7 gene).

[0147] The disease or condition may have symptoms of cell overgrowth. For example, the disease or condition may be cancer (for example, cancer in which CCNE1 is overexpressed or has an inactivating mutation in the FBXW7 gene).

[0148] Examples of cancers with a high incidence of CCNE1 overexpression include, for example, uterine cancer, ovarian cancer, breast cancer, gastric cancer, esophageal cancer, lung cancer, and endometrial cancer.

[0149] Cancers with FBXW7 deficiency include, for example, uterine cancer, colon cancer, breast cancer, lung cancer, and esophageal cancer.

[0150] The compounds of the present invention can be administered by a route selected from the group consisting of oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intraarterial, intracranial, subcutaneous, intraorbital, intraventricular, intraspinal, intraperitoneal, intranasal, inhalation, intratumoral, and topical administration.

[0151] Pharmaceutical Composition The compounds used in the methods described herein are preferably formulated into pharmaceutical compositions for administration to human subjects in a biologically compatible form suitable for in vivo administration. Pharmaceutical compositions typically include a compound described herein and a pharmaceutically acceptable excipient. Certain pharmaceutical compositions may also include one or more additional pharmaceutically active agents described herein.

[0152] The compounds described herein can also be used in the form of a free base, a salt, a zwitterion, a solvate, or a prodrug, or a pharmaceutical composition thereof. All forms are within the scope of the present invention. As will be understood by those skilled in the art, the compounds, salts, zwitterions, solvates, prodrugs, or pharmaceutical compositions thereof can be administered to patients in various forms depending on the selected route of administration. The compounds used in the methods described herein can be administered, for example, orally, parenterally, bucally, sublingually, nasally, rectally, via a patch, via a pump, or transdermally, and pharmaceutical compositions formulated accordingly. Parenteral administration includes intravenous, intraperitoneal, subcutaneous, intramuscular, transepithelial, nasal, pulmonary, intrathecal, rectal, and topical modes of administration. Parenteral administration can be by continuous infusion over a selected period of time.

[0153] For human use, the compounds of the present invention can be administered alone or in combination with a pharmaceutical carrier selected with respect to the intended route of administration and standard medical practice. Accordingly, pharmaceutical compositions according to the present invention can be formulated in conventional manner using one or more pharmaceutically acceptable carriers which facilitate the processing of the compounds of the present invention into preparations which can be used pharmaceutically and include excipients and auxiliaries which are suitable for pharmaceutical use.

[0154] The present invention also includes pharmaceutical compositions which may contain one or more pharmaceutically acceptable carriers. In the manufacture of the pharmaceutical compositions of the present invention, the active ingredient is usually admixed with, diluted by, or enclosed within a carrier such as, for example, in the form of capsules, sachets, paper, or other containers. When the excipient functions as a diluent, it can be a solid, semi-solid, or liquid substance (such as normal saline) which acts as a vehicle, carrier, or medium for the active ingredient. Accordingly, the compositions can be in the form of tablets, powders, lozenges, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, and soft and hard gelatin capsules. As is known in the art, the type of diluent can vary depending on the intended route of administration. The resulting compositions may contain additional agents, for example, preservatives.

[0155] The excipient or carrier is selected based on the mode and route of administration. Suitable pharmaceutical carriers, as well as the pharmaceutical necessities used in pharmaceutical formulations, are described in the well-known reference books in the art, Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005), and USP / NF (United States Pharmacopeia and National Formulary). Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose. The formulations may further include lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl benzoate and propyl hydroxybenzoate; sweetening agents; and flavoring agents. Other exemplary excipients are described in Handbook of Pharmaceutical Excipients, 6th Edition, Rowe et al., Eds., Pharmaceutical Press (2009).

[0156] These pharmaceutical compositions can be manufactured by conventional methods, for example, by processes such as conventional mixing, dissolving, granulating, tablet coating, powdering, emulsifying, encapsulating, entrapping, or lyophilizing. Methods for manufacturing formulations well-known in the art are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005), and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988 - 1999, Marcel Dekker, New York. Suitable formulations depend on the selected route of administration. The formulation and preparation of such compositions are well-known to those skilled in the pharmaceutical formulation art. When preparing a formulation, the active compound can be milled to an appropriate particle size before being formulated with other ingredients. If the active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh. If the active compound is substantially water-soluble, the particle size can be adjusted, for example, to about 40 mesh by milling and dispersed almost uniformly in the formulation.

[0157] Dosage The dosage of the compounds, or pharmaceutically acceptable salts or prodrugs thereof, or pharmaceutical compositions thereof, used in the methods described herein can vary depending on many factors, such as the pharmacodynamic properties of the compound; the method of administration; the age, health status, and weight of the recipient; the nature and extent of the symptoms; the frequency of treatment, and in the case of co-treatment, the type of co-treatment; and the clearance rate of the compound in the animal being treated. One of ordinary skill in the art can determine an appropriate dosage based on the above factors. The compounds used in the methods described herein can be administered initially at an appropriate dosage and the dosage can be adjusted as needed depending on the clinical response. Generally, an appropriate daily dosage of the compounds of the present invention is considered to be the amount of the compound that is the lowest effective dosage to produce a therapeutic effect. Such effective dosages will generally vary depending on the above factors.

[0158] The compounds of the present invention can be administered to a patient in a single dose or multiple doses. When administering multiple doses, each dose may be divided from each other, for example, every 1 to 24 hours, 1 to 7 days, 1 to 4 weeks, or 1 to 12 months. The compound can be administered according to a schedule or without setting a predetermined schedule. The active compound can be administered, for example, once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, eleven times, or twelve times a day, every two days, every three days, every four days, every five days, or every six days, once, twice, three times, four times, five times, six times, or seven times a week, once, twice, three times, four times, five times, or six times a month, or once, twice, three times, four times, five times, six times, seven times, eight times, nine times, ten times, eleven times, or twelve times a year. It should be understood that the specific dosage regimen for a particular subject needs to be adjusted over time according to the individual in need and the professional judgment of the individual who manages or directs the administration of the composition.

[0159] Ultimately, the attending physician will determine the appropriate amount and dosage regimen, but the effective amount of the compounds of the present invention can be, for example, the total daily dose, for example, any of the compounds described herein from 0.05 mg to 3000 mg. Alternatively, the dosage can be calculated using the patient's body weight. Such dosage ranges can include, for example, 10 to 1000 mg (for example, 50 to 800 mg). In some embodiments, 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg of the compound is administered.

[0160] In the method of the present invention, the period during which multiple doses of the compound of the present invention are administered to a patient can vary. For example, in some embodiments, the dose of the compound of the present invention is administered to the patient over a period of 1 to 7 days; 1 to 12 weeks; or 1 to 3 months. In some embodiments, the compound is administered to the patient over a period of, for example, 4 to 11 months or 1 to 30 years. In some embodiments, the compound is administered to the patient at the onset of symptoms. In any of such embodiments, the amount of the compound to be administered may be changed during the administration period. When the compound is administered daily, for example, it can be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times a day.

[0161] Formulation A compound identified to be able to treat any of the conditions described herein using any of the methods described herein can be administered to a patient or animal in unit dosage form together with a pharmaceutically acceptable diluent, carrier, or excipient. The chemical compounds for use in such therapies can be manufactured and isolated by any standard technique known to those skilled in the art of pharmaceutical chemistry. Suitable formulations or compositions for administering the identified compound to a patient suffering from a disease or condition can be provided using conventional pharmaceutical practices. Administration may be initiated before the patient shows symptoms.

[0162] Exemplary routes of administration of the compound (e.g., the compound of the present invention) or its pharmaceutical composition used in the present invention include oral, sublingual, buccal, transdermal, intradermal, intramuscular, parenteral, intravenous, intraarterial, intracranial, subcutaneous, intraorbital, intraventricular, intraspinal, intraperitoneal, intranasal, inhalation, and topical administration. The compound is preferably administered together with a pharmaceutically acceptable carrier. The pharmaceutical formulations of the compounds described herein formulated for treating the disorders described herein are also part of the present invention.

[0163] Formulations for oral administration The pharmaceutical compositions contemplated by the present invention include those formulated for oral administration ("oral dosage forms"). Oral dosage forms can be, for example, tablets, capsules, solutions or suspensions, powders, or in the form of liquid crystals or solid crystals, which contain the active ingredient(s) in a mixture with non-toxic pharmaceutically acceptable excipients. Such excipients can be, for example, inert diluents or bulking agents (e.g., sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starch including potato starch, calcium carbonate, sodium chloride, lactose, calcium phosphate, calcium sulfate, or sodium phosphate); granulating and disintegrating agents (e.g., cellulose derivatives including microcrystalline cellulose, starch including potato starch, croscarmellose sodium, alginates, or alginic acid); binders (e.g., sucrose, glucose, sorbitol, acacia, alginic acid, sodium alginate, gelatin, starch, pregelatinized starch, microcrystalline cellulose, magnesium aluminum silicate, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropylmethyl cellulose, ethyl cellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricants, glidants, and antiadhesion agents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharmaceutically acceptable excipients can be coloring agents, flavoring agents, plasticizers, wetting agents, buffering agents, and the like.

[0164] Formulations for oral administration can also be in the form of chewable tablets, hard gelatin capsules in which an inert solid diluent (e.g., potato starch, lactose, microcrystalline cellulose, calcium carbonate, calcium phosphate, or kaolin) is mixed with the active ingredient, or soft gelatin capsules in which water or an oily medium, such as peanut oil, liquid paraffin, or olive oil, is mixed with the active ingredient. Powders, granules, and pellets can be prepared by conventional methods using the ingredients described above for tablets and capsules, for example, using a mixer, a fluidized bed apparatus, or a spray drying apparatus.

[0165] Oral controlled-release compositions can be constructed to control the dissolution and / or diffusion of the active pharmaceutical ingredient to release the active drug. To achieve controlled release and the targeted plasma concentration-time profile, any of several means may be employed. In one example, controlled release is obtained by appropriate selection of various formulation parameters and components, including, for example, various types of controlled-release compositions and coatings. Examples include single-unit or multiple-unit tablet or capsule compositions, oils, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and liposomes. In some embodiments, the composition includes a biodegradable, pH-, and / or temperature-sensitive polymer coating.

[0166] Dissolution or diffusion-controlled release can be achieved by appropriate coating of tablets, capsules, pellets, or granules of the compound, or by incorporation of the compound into an appropriate matrix. Controlled-release coatings can include one or more of the coating substances described above and / or, for example, shellac, beeswax, glyceryl wax, castor oil wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glyceryl palmitostearate, ethyl cellulose, acrylic resin, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinyl pyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-hydroxy methacrylate, methacrylate hydrogel, 1,3 butylene glycol, ethylene glycol methacrylate, and / or polyethylene glycol. In controlled-release matrix formulations, the matrix material can also include, for example, hydrated methyl cellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and / or halogenated fluorocarbon.

[0167] As liquid forms into which the compounds and compositions of the present invention can be incorporated in the case of oral administration, there may be mentioned aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and flavored emulsions using edible oils (e.g., cottonseed oil, sesame oil, coconut oil, or peanut oil), as well as elixirs and similar pharmaceutical vehicles.

[0168] Formulations for parenteral administration The compounds described herein for use in the methods of the present invention can be administered in the pharmaceutically acceptable parenteral (e.g., intravenous or intramuscular) formulations described herein. The pharmaceutical formulations can also be administered parenterally (intravenously, intramuscularly, subcutaneously, etc.) in dosage forms or formulations containing conventional non-toxic pharmaceutically acceptable carriers and adjuvants. In particular, as formulations suitable for parenteral administration, there may be mentioned aqueous and non-aqueous sterile injection solutions which may contain antioxidants, buffers, bacteriostats, and solutes which render the formulation isotonic with the blood of the intended recipient, and aqueous and non-aqueous sterile suspensions which may contain suspending and thickening agents. For example, to prepare such compositions, the compounds of the present invention can be dissolved or suspended in a parenterally acceptable liquid vehicle. Acceptable vehicles and solvents that can be used include water, water adjusted to an appropriate pH with an appropriate amount of hydrochloric acid, sodium hydroxide, or an appropriate buffer, 1,3-butanediol, Ringer's solution, and isotonic sodium chloride solution. Aqueous formulations can also contain one or more preservatives, such as methyl, ethyl, or n-propyl p-hydroxybenzoate. Additional information regarding parenteral formulations can be found, for example, in the United States Pharmacopeia-National Formulary (USP-NF), which is incorporated herein by reference.

[0169] Parenteral formulations are five general types of preparations confirmed by the USP-NF to be suitable for parenteral administration, namely (1) "Drug injection solution": a liquid preparation that is the active ingredient (e.g., the compound of the present invention), or a solution thereof; (2) "Drug for injection": the active ingredient (e.g., the compound of the present invention) as a dry solid combined with a sterile vehicle suitable for parenteral administration as a drug injection solution; (3) "Emulsion for drug injection": A liquid preparation of a drug substance (e.g., a compound of the present invention) dissolved or dispersed in a suitable emulsion medium; (4) "Suspension for drug injection": A liquid preparation of a drug substance (e.g., a compound of the present invention) suspended in a suitable liquid medium; (5) "Drug for suspension for injection": Any of the drug substances (e.g., a compound of the present invention) in the form of a dry solid that is combined with a sterile vehicle suitable for parenteral administration as a suspension for drug injection.

[0170] Examples of formulations for parenteral administration include aqueous solutions of compounds prepared by appropriately mixing surfactants, such as hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycol, DMSO, and mixtures thereof (with or without alcohol), as well as in oils. Under normal storage and use conditions, these preparations may contain preservatives to prevent the growth of microorganisms. Conventional procedures and components for the selection and preparation of suitable formulations are described, for example, in Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005) and The United States Pharmacopeia: The National Formulary (USP 36 NF31), published in 2013.

[0171] Formulations for parenteral administration may contain, for example, excipients such as sterile water or saline, polyalkylene glycols such as polyethylene glycol, vegetable oils, or hydrogenated naphthalene. Biocompatible, biodegradable lactide polymers, lactide / glycolide copolymers, or polyoxyethylene-polyoxypropylene copolymers can be used to control the release of the compounds. Other potentially useful parenteral compound delivery systems include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Inhalation formulations may contain excipients such as lactose, or may be aqueous solutions containing, for example, polyoxyethylene-9-lauryl ether, glycocholate, and deoxycholate, or oily solutions for administration in the form of nasal drops or as gels.

[0172] Parenteral formulations can be formulated for immediate release or sustained / extended release of the compound. Exemplary formulations for parenteral release of the compound include aqueous solutions, powders for reconstitution, co-solvent solutions, oil / water emulsions, suspensions, oil-based solutions, liposomes, microspheres, and polymer gels.

[0173] combination The compounds of the present invention may be administered in combination with one or more additional agents, e.g. (a) cytotoxic agents; (b) antimetabolites; (c) alkylating agents; (d) anthracyclines; (e) antibiotics; (f) antimitotic agents; (g) hormone therapy; (h) signal transduction inhibitors; (i) Gene expression regulators; (j) apoptosis inducers; (k) angiogenesis inhibitors; (l) immunotherapeutic agents; (m) DNA damage repair inhibitors; or combinations of these can be administered to a subject in combination with

[0174] Cytotoxic agents include, for example, actinomycin-D, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, amphotericin, amsacrine, arsenic trioxide, asparaginase, azacitidine, azathioprine, Bacillus Calmette-Guerin (BCG), bendamustine, bexarotene, bevacizumab, bleomycin, bortezomib, busulfan, capecitabine, carboplatin, carfilzomib, carmustine, cetuximab, cisplatin, chlorambucil, cladribine, clofarabine, colchicine, crisantasparase, cyclophosphamide, cyclosporine, cytarabine, cytochalasin B, dacarbazine, dactinomycin, darbepoetin alfa, dasatinib, daunorubicin, 1-dehydrotestosterone, denileukin, dexamethasone, dexrazoxane, dihydroxyanthracenedione, disulfiram, docetaxel, doxorubicin, emetine, epirubicin, erlotinib, epigallocatechin gallate, epoetin alfa, estramustine, ethidium bromide, etoposide, everolimus, filgrastim, finasteride, floxuridine, fludarabine, fluorouracil (5-FU), fulvestrant, ganciclovir, geldanamycin, gemcitabine, glucocorticoid, gramicidin D, histrelin acetate, hydroxyurea, ibritumomab, idarubicin, ifosfamide, imatinib, irinotecan, interferon, interferon alpha-2a, interferon alpha-2b, ixabepilone, lactate dehydrogenase A (LDH-A), lenalidomide, letrozole, leucovorin, levamisole, lidocaine, lomustine, mechlorethamine, melphalan, 6-mercaptopurine, mesna, methotrexate, methoxsalen, methopterin, metronidazole, mitramycin, mitomycin-C, mitoxantrone, nandrolone, nelarabine, nilotinib, nolatrexed, oprelvekin, oxaliplatin, paclitaxel, pemetrexed, pentostatin, parifermin, pamidronic acid, pegademase, pegaspargase, pegfilgrastim, pemetrexed disodium, plicamycin, porfimer sodium, procaine, procarbazine, propranolol,The agent may be puromycin, quinacrine, radicicol, a radioisotope, raltitrexed, rapamycin, rasburicase, salinosporamide A, sargramostim, sunitinib, temozolomide, teniposide, tetracaine, 6-thioguanine, thiotepa, topotecan, toremifene, trastuzumab, treosulfan, tretinoin, valrubicin, vinblastine, vincristine, vindesine, vinorelbine, zoledronate, or a combination thereof.

[0175] The antimetabolite can be, for example, methotrexate, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-fluorouracil decarbazine, cladribine, pemetrexed, gemcitabine, capecitabine, hydroxyurea, mercaptopurine, fludarabine, pralatrexate, clofarabine, cytarabine, decitabine, floxuridine, nelarabine, trimetrexate, thioguanine, pentostatin, or a combination thereof.

[0176] The alkylating agent can be, for example, mechlorethamine, thiotepa, chlorambucil, melphalan, carmustine (BSNU), lomustine (CCNU), cyclophosphamide, busulfan, dibromomannitol, streptozotocin, mitomycin C, cis-dichlorodiamineplatinum(II) (DDP) cisplatin, altretamine, cyclophosphamide, ifosfamide, hexamethylmelamine, altretamine, procarbazine, dacarbazine, temozolomide, streptozotocin, carboplatin, cisplatin, oxaliplatin, uramustine, bendamustine, trabectedin, semustine, or a combination thereof.

[0177] The anthracycline can be, for example, daunorubicin, doxorubicin, acarrubicin, aldoxorubicin, amrubicin, annamycin, carubicin, epirubicin, idarubicin, mitoxantrone, valrubicin, or a combination thereof.

[0178] Antibiotics include, for example, dactinomycin, bleomycin, mithramycin, anthramycin (AMC), ampicillin, bacampicillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, nafcillin, oxacillin, piperacillin, pivampicillin, pivmecillinam, ticarcillin, aztreonam, imipenem, doripenem, ertapenem, meropenem, cephalosporins, clarithromycin, dirithromycin, roxithromycin, telithromycin, lincomycin, pristinamycin, quinupristin, amikacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, tobramycin, streptomycin, sulfamethizole, sulfamethoxazole, sulfisoxazole, demecloxacillin, The anti-inflammatory drug may be iclin, minocycline, oxytetracycline, tetracycline, penicillin, amoxicillin, cephalexin, erythromycin, clarithromycin, azithromycin, ciprofloxacin, levofloxacin, ofloxacin, doxycycline, clindamycin, metronidazole, tigecycline, chloramphenicol, metronidazole, tinidazole, nitrofurantoin, vancomycin, teicoplanin, telavancin, linezolid, cycloserine, rifamycin, polymyxin B, bacitracin, viomycin, capreomycin, quinolone, daunorubicin, doxorubicin, 4'-deoxydoxorubicin, epirubicin, idarubicin, plicamycin, mitomycin-c, mitoxantrone, or a combination thereof.

[0179] The antimitotic agent can be, for example, vincristine, vinblastine, vinorelbine, docetaxel, estramustine, ixabepilone, paclitaxel, maytansinoids, dolastatins, cryptophycins, or combinations thereof.

[0180] The signal transduction inhibitor can be, for example, imatinib, trastuzumab, erlotinib, sorafenib, sunitinib, temsirolimus, vemurafenib, lapatinib, bortezomib, cetuximab, panitumumab, matuzumab, gefitinib, STI 571, rapamycin, flavopiridol, imatinib mesylate, vatalanib, semaxinib, motesanib, axitinib, afatinib, bosutinib, crizotinib, cabozantinib, dasatinib, entrectinib, pazopanib, lapatinib, vandetanib, or a combination thereof.

[0181] Gene expression regulator can be, for example, siRNA, shRNA, antisense oligonucleotide, HDAC inhibitor, or a combination thereof.HDAC inhibitor can be, for example, trichostatin A, trapoxin B, valproic acid, vorinostat, belinostat, LAQ824, panobinostat, entinostat, tacedinaline, mocetinostat, gibinostat, resminostat, abexinostat, xinostat, rosirinostat, pracinostat, CHR-3996, butyric acid, phenylbutyric acid, 4SC202, romidepsin, sirtinol, cambinol, EX-527, nicotinamide, or a combination thereof.Antisense oligonucleotide can be, for example, custorisen, apatursen, AZD9150, travedersen, EZN-2968, LErafAON-ETU, or a combination thereof. The siRNA can be, for example, ALN-VSP, CALAA-01, Atu-027, SPC2996, or a combination thereof.

[0182] Hormone therapy can be, for example, a luteinizing hormone-releasing hormone (LHRH) antagonist. Hormone therapy can be, for example, Pharmagon, leuprorelin, goserelin, buserelin, flutamide, bicalutamide, ketoconazole, aminoglutethimide, prednisone, hydroxyprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, diethylstilbestrol, ethinyl estradiol, tamoxifen, testosterone propionate, fluoxymesterone, flutamide, raloxifene, droloxifene, iodoxyfene, 4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, tamoxifen citrate, megestrol acetate, exemestane, fadrozole, vorozole, letrozole, anastrozole, nilutamide, tripterelin, histrelin, abiraterone, medroxyprogesterone acetate, diethylstilbestrol, Premarin, fluoxymesterone, tretinoin, fenretinide, troxacitabine, or a combination thereof.

[0183] The apoptosis inducer can be, for example, recombinant human TNF-related apoptosis-inducing ligand (TRAIL), camptothecin, bortezomib, etoposide, tamoxifen, or a combination thereof.

[0184] The angiogenesis inhibitor can be, for example, sorafenib, sunitinib, pazopanib, everolimus, or a combination thereof.

[0185] The immunotherapeutic agent may be, for example, a monoclonal antibody, a cancer vaccine (e.g., a dendritic cell (DC) vaccine), an oncolytic virus, a cytokine, adoptive T cell therapy, Mycobacterium Calmette-Guérin (BCG), GM-CSF, thalidomide, lenalidomide, pomalidomide, imiquimod, or a combination thereof. The monoclonal antibody may be, for example, anti-CTLA4, anti-PD1, anti-PD-L1, anti-LAG3, anti-KIR, or a combination thereof. The monoclonal antibody can be, for example, alemtuzumab, trastuzumab, ibritumomab tiuxetan, brentuximab vedotin, trastuzumab, adotrastuzumab emtansine, blinatumomab, bevacizumab, cetuximab, pertuzumab, panitumumab, ramucirumab, obinutuzumab, ofatumumab, rituximab, pertuzumab, tositumomab, gemtuzumab, ozogamicin, tositumomab, or a combination thereof. The cancer vaccine may be, for example, Sipuleucel-T, BioVaxID, NeuVax, DCVax, SuVaxM, CIMAvax®, Provenge®, hsp110 chaperone complex vaccine, CDX-1401, MIS416, CDX-110, GVAX pancreatic vaccine, HyperAcute™ pancreatic vaccine, GTOP-99 (MyVax®), or Imprime PGG®. The oncolytic virus may be, for example, talimogene laherparepvec. The cytokine may be, for example, IL-2, IFNα, or a combination thereof. The adoptive T cell therapy may be, for example, tisagenlecleucel, axicabtageneciloreucel, or a combination thereof.

[0186] DNA damage repair inhibitors can be, for example, PARP inhibitors, cell checkpoint kinase inhibitors, or combinations thereof. PARP inhibitors can be, for example, olaparib, rucaparib, veliparib (ABT-888), niraparib (ZL-2306), iniparib (BSI-201), talazoparib (BMN 673), 2X-121, CEP-9722, KU-0059436 (AZD2281), PF-01367338, or combinations thereof. Cell checkpoint kinase inhibitors can be, for example, MK-1775 or AZD1775, AZD7762, LY2606368, PF-0477736, AZD0156, GDC-0575, ARRY-575, CCT245737, PNT-737, or combinations thereof.

Example

[0187] The following examples are intended to illustrate the present invention. They are in no way intended to limit the present invention. In the following examples, the reactions were typically carried out at room temperature (rt) under a nitrogen atmosphere using an anhydrous solvent (Sure / Seal (trademark)), unless otherwise stated. TLC was performed or a small aliquot was injected into a Waters Acquity-H UPLC (registered trademark) Class system using an Acquity (registered trademark) UPLC HSS C18 2.1×30 mm column, and eluted with a gradient (1.86 minutes) of acetonitrile water (both containing 0.1% formic acid) from 15% to 98% to follow the reaction. Purification by preparative HPLC was carried out in a Teledyne Isco Combi Flash (registered trademark) EZ Prep system at a flow rate of 40 mL / min for 12 minutes using a Phenomenex Gemini (registered trademark) 5μm NX-C18 110Å 150×21.2 mm column (less than 100 mg, or multiple injections of less than 100 mg), or using an HP C18 RediSep (registered trademark) Rf gold column (more than 100 mg), eluting with an appropriate gradient of acetonitrile water (both containing 0.1% formic acid). The gradient was selected based on the retention times measured by following the reaction in a Waters Acquity-H UPLC (registered trademark) Class system (see above). Fractions containing the desired compound were combined and finally lyophilized. Purification by silica gel chromatography was carried out in a Teledyne Isco Combi Flash (registered trademark) Rf system using a RediSep (registered trademark) Rf silica gel column of appropriate size. The purity of the final compound was evaluated by injecting a small aliquot into a Waters Acquity-H UPLC (registered trademark) Class system using an Acquity (registered trademark) UPLC BEH C18 2.1×50 mm column and eluting with a gradient (7 minutes) of acetonitrile water (both containing 0.1% formic acid) from 2% to 98%.

[0188] Abbreviations Abbreviations and terms commonly used in the fields of organic chemistry, pharmaceutical chemistry, pharmacology, and medicine and well-known to experts in these fields are used herein. Representative abbreviations and definitions are shown below: Ac is acetyl [CH3C(O)-]; ACN is acetonitrile; Ac2O is acetic anhydride; AcOH is acetic acid; Ar is aryl; BOC is tert-butyloxycarbonyl; n-BuLi is n-butyllithium; cmpd is compound; conc. is concentrated; DCM is dichloromethane; DIPEA is diisopropylethylamine; DMAP is 4-(dimethylamino)pyridine; DME is dimethoxyethane; DMF is N,N-dimethylformamide; DMSO is dimethyl sulfoxide; EtOAc is ethyl acetate; EtOH is ethanol; h is hour; HATU is 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate; HCl is hydrochloric acid; Hex is hexane; HPLC is high performance liquid chromatography; IPA is isopropanol; LCMS is HPLC with mass spectrometry detection; LiHMDS is lithium hexamethyldisilazane; M is molarity and mmol is millimole; Me is methyl; MeCN is acetonitrile; MeMgBr is methylmagnesium bromide; MeMgCl is methylmagnesium chloride; MeOH is methanol; MOM is methoxymethyl; min is minute; N is normal; NBS is N-bromosuccinimide; NCS is N-chlorosuccinimide; NIS is N-iodosuccinimide; NMP is N-methylpyrrolidine; NMR is nuclear magnetic resonance spectroscopy; PdCl2(dppf) is [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II); PdCl2(dppf).CH2Cl2 is a complex of [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) and dichloromethane; Pd2(dba)3 is tris(dibenzylideneacetone)dipalladium(0); Pd-PEPPSI™-SIPr is (1,3-bis(2,6-diisopropylphenyl)imidazolidene)(3-chloropyridyl)palladium(II) dichloride; Ph is phenyl; PIV-Cl is pivaloyl chloride, trimethylacetyl chloride; Reagent alcohol is a mixture of 90% ethanol, 5% isopropanol and 5% methanol; rt is room temperature; sat. is saturated; tBu is tert-butyl; Tf is trifluoromethanesulfonate; TFA is trifluoroacetic acid; THF is tetrahydrofuran; TMS is trimethylsilyl; Ts is p-toluenesulfonyl; Xantphos is 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene.

[0189] Example 1. Preparation of Compounds

Chemical Structure

[0190] Step 2. Benzyl bromide (48 mL, 404 mmol) was added dropwise to a suspension of 5-bromo-6-chloro-pyrazin-2-ol (80 g, 382 mmol) and silver carbonate (216 g, 778 mmol) in toluene (2 L). After stirring for 3 hours, the suspension was filtered through Celite. The filtrate was evaporated to dryness to obtain a yellow oil, which was dissolved in warm EtOH. After slowly adding water under sonication, the precipitate was collected by filtration to obtain 5-benzyloxy-2-bromo-3-chloro-pyrazine (85.2 g, 75% yield) as a pale yellowish brown solid. 1 H NMR (400 MHz, DMSO-d6) δ 8.27 (s, 1H), 7.51 - 7.46 (m, 2H), 7.44 - 7.33 (m, 3H), 5.36 (s, 2H).

[0191]

Chemical Structure

[0192] Step 2. To a solution of propanedinitrile (42.1 g, 637 mmol) in DME (1800 mL) was added NaH (25.0 g, 628 mmol, 60% dispersion in mineral oil) portionwise in several times. The resulting mixture was stirred for 30 minutes, and then DME (500 mL) containing 5-benzyloxy-3-chloro-N-(3-methoxy-2,6-dimethyl-phenyl)pyrazine-2-amine (115 g, 311 mmol) and Pd(PPh3)4 (17.7 g, 15.3 mmol) were added. The resulting mixture was stirred under reflux for 2 hours and then concentrated to 1 L under vacuum. Water (1 L) was slowly added and the resulting biphasic mixture was stirred with a mechanical stirrer for 18 hours. The resulting solid was removed by filtration, washed with water and dried under vacuum. The desired substance of the first batch was obtained as a pale yellowish brown solid isolated by filtration after trituration in DCM. The mother liquor was concentrated under vacuum and the residue was purified by silica gel chromatography (dry packing) eluting with a gradient of 10 - 60% EtOAc in hexane to give the desired substance of the second batch. The two batches were combined to give 6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (103.1 g, 83% yield) as a pale yellowish brown solid. 1 1H NMR (400 MHz, chloroform-d) δ 7.60 (s, 1H), 7.53 - 7.47 (m, 2H), 7.42 - 7.34 (m, 2H), 7.33 - 7.27 (m, 1H), 7.21 - 7.15 (m, 1H), 6.94 (d, J = 8.5 Hz, 1H), 5.45 (s, 2H), 4.91 (s, 2H), 3.84 (s, 3H), 1.90 (d, J = 0.7 Hz, 3H), 1.83 (s, 3H). MS: [M+1]: 400.4.

[0193]

Chem.

[0194] Step 2. To a solution of 6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (30.5 g, 93.2 mmol) and CsCO (34.9 g, 107 mmol) in DMF (300 mL) was added 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (36.6 g, 103 mmol). The reaction mixture was stirred for 1 h, diluted with water (900 mL), and extracted with EtOAc (3 × 300 mL). The combined organic extracts were washed with water, brine, dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 20 to 100% EtOAc in hexanes to give [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (28 g, 65% yield) as an off-white solid. 1 H NMR (400 MHz, chloroform-d) δ 7.75 (s, 1H), 7.22 m, 2H), 6.97 (d, J = 8.5 Hz, 1H), 6.37 (s, 2H), 5.49 (s, 1H), 3.86 (s, 3H), 1.91 (s, 3H), 1.84 (s, 3H). MS: [M+1]: 528.4.

[0195] Chiral SFC separation of intermediate D (7.0 g, 15 mmol) (apparatus: Waters Prep 100 SFC-MS; column: Phenomenex Lux Cellulose-2, 30×250 mm, 5 μm; conditions: 75% CO2, isocratic with 25% MeOH; flow rate: 70 mL / min) gave intermediate D1 and intermediate D2.

Chem.

Chem.

[0196] [ka] Intermediate E (6-amino-3-hydroxy-5-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile) Step 1. To a solution of 5-benzyloxy-2-bromo-3-chloro-pyrazine (5.08 g, 17.0 mmol) and 5-(methoxymethoxy)-2-methyl-aniline (5.70 g, 34.1 mmol) in THF (40 mL) at 0 °C, potassium tert-butoxide in THF (1 M, 48 mL) was added dropwise. After stirring for 90 min at 0 °C, the reaction mixture was quenched with saturated NH4Cl, diluted with water, and extracted with EtOAc (3x). The combined organic extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of 0–20% EtOAc in hexanes to give 6-benzyloxy-3-bromo-N-[5-(methoxymethoxy)-2-methyl-phenyl]pyrazin-2-amine (1.80 g, 25% yield) as a pale yellow solid.

[0197] Step 2. To a suspension of NaH (631 mg, 16.5 mmol, 60% dispersion in mineral oil) in THF (28 mL) at 0 °C, malononitrile (556 mg, 8.42 mmol) in THF (12 mL) was added dropwise. After stirring at 0 °C for 30 min, the ice bath was removed and 6-benzyloxy-3-bromo-N-[5-(methoxymethoxy)-2-methyl-phenyl]pyrazin-2-amine (1.80 g, 4.18 mmol) and Pd(PPh3)4 (242 mg, 209 μmol) were added. The resulting mixture was flushed with nitrogen and stirred at 60 °C for 1 h. The resulting mixture was cooled to room temperature and slowly poured into saturated aqueous NH4Cl solution, then extracted with EtOAc (2x). The combined organic extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (dry packing) eluting with a gradient of 0 to 100% EtOAc in hexanes to give 6-amino-3-benzyloxy-5-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile (1.63 g, 94% yield) as a light tan solid.

[0198] Step 3. A mixture of 6-amino-3-benzyloxy-5-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile (1.39 g, 3.35 mmol) and palladium on carbon (350 mg, 0.329 mmol, 10% w / w) was flushed with nitrogen and MeOH (40 mL) was added. The reaction mixture was flushed with hydrogen and stirred under a hydrogen atmosphere (1 atm) for 2 hours, then flushed with nitrogen and filtered through a Celite pad using DCM and MeOH. The filtrate was concentrated in vacuo and then dried to give 6-amino-3-hydroxy-5-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile (1.12 g, 100%) as an ochre solid. MS: [M+1]: 326.1.

[0199] [ka] Intermediate F ([6-amino-7-cyano-5-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[2,3-b]pyrazin-3-yl] trifluoromethanesulfonate) Step 1. To a solution of 6-amino-3-hydroxy-5-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile (1.12 g, 3.44 mmol) and 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (1.49 g, 4.17 mmol) in THF (45 mL) was added EtN (1.23 g, 12.2 mmol, 1.70 mL). The reaction mixture was stirred for 18 h and then concentrated in vacuo. The residue was purified by silica gel chromatography (dry packed) eluting with a gradient of 0 to 100% EtOAc in hexanes to give [6-amino-7-cyano-5-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[2,3-b]pyrazin-3-yl] trifluoromethanesulfonate (1.72 g, 100%) as a dark yellow solid. 1 H NMR(400MHz,DMSO-d6)δ8.38(s,1H),8.13(br s,2H),7.41(dd,J=8.5,0.9Hz,1H),7.19(dd,J=8.5,2.6Hz,1H),7.11(d,J=2.6 Hz,1H),5.22(d,J=6.8Hz,1H),5.17(d,J=6.8Hz,1H),3.38(s,3H),1.90(s,3H). MS:[M+1]:458.0.

[0200] [ka] Intermediate G (2-amino-5-chloro-1-(5-hydroxy-2-methyl-phenyl)pyrrolo[3,2-b]pyridine-3-carboxamide) Step 1. To a solution of propanedinitrile (11.8 g, 179 mmol) in DME (200 mL) at 0 °C was added NaH (7.0 g, 175.00 mmol, 60% dispersion in mineral oil) portionwise in several times. Then, 3-bromo-2,6-dichloro-pyridine (20 g, 88.15 mmol) was added and the resulting mixture was stirred and stirred at 90 °C for 6 h. The reaction mixture was cooled to room temperature, neutralized with 1 M HCl, diluted with water, and extracted with EtOAc (3×). The combined organic extracts were washed with brine, filtered, and concentrated under vacuum. The residue was purified by preparative HPLC in several batches. The desired fractions were combined and concentrated to dryness to give 2-(3-bromo-6-chloro-2-pyridyl)propanedinitrile (8.0 g, 35% yield) as an off-white solid.

[0201] Step 2. To a solution of 2-(3-bromo-6-chloro-2-pyridyl)propanedinitrile (5 g, 19.5 mmol) in DMF (75 mL) were added Pd2(dba)3 (1.75 g, 1.91 mmol), 5-(methoxymethoxy)-2-methyl-aniline (3.6 g, 21.53 mmol), Cs2CO3 (12.7 g) and Xantphos (1.12 g, 1.94 mmol). The mixture was degassed under vacuum and refilled with nitrogen three times. The resulting mixture was stirred at 130 °C for 8 h and then cooled to room temperature. The resulting mixture was diluted with water and extracted with EtOAc (3×). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 10 - 60% EtOAc in hexane. The desired fractions were concentrated to dryness and the residue was triturated with DCM to give 2-amino-5-chloro-1-[5-(methoxymethoxy)-2-methyl-phenyl]pyrrolo[3,2-b]pyridine-3-carbonitrile (2.2 g, 33% yield) as an off-white solid.

[0202] Step 3. To a suspension of 3.2 - amino - 5 - chloro - 1 - [5 - (methoxymethoxy) - 2 - methyl - phenyl] pyrrolo[3,2 - b] pyridine - 3 - carbonitrile (2.20 g, 6.42 mmol) in DCM (5 mL) was added hydrogen chloride 4 M in dioxane (4 M, 5 mL). The mixture was stirred for 30 minutes. Volatiles were removed under vacuum to afford 2 - amino - 5 - chloro - 1 - (5 - hydroxy - 2 - methyl - phenyl) pyrrolo[3,2 - b] pyridine - 3 - carbonitrile HCl salt (2.10 g, 98% yield) as an off - white solid.

[0203] Step 4. A solution of 2 - amino - 5 - chloro - 1 - (5 - hydroxy - 2 - methyl - phenyl) pyrrolo[3,2 - b] pyridine - 3 - carbonitrile (2.3 g, 7.70 mmol) in concentrated sulfuric acid (25 mL) was stirred at room temperature for 1 hour. It was then diluted with crushed ice and basified to pH 8 with concentrated aqueous ammonia. The suspension was filtered. The precipitate was washed with water and dried under vacuum to give an off - white mixture mainly containing 2 - amino - 5 - chloro - 1 - (5 - hydroxy - 2 - methyl - phenyl) pyrrolo[3,2 - b] pyridine - 3 - carboxamide (2 g, 82% yield), which was used directly in the next step without further purification. 1 H NMR (400 MHz, DMSO - d6) δ 9.77 (s, 1H), 7.42 (s, 1H), 7.30 (d, J = 8.3 Hz, 1H), 7.16 (m, 3H), 6.98 - 6.91 (m, 2H), 6.87 (d, J = 8.1 Hz, 1H), 6.71 (d, J = 2.6 Hz, 1H), 1.81 (s, 3H). MS: [M + 1]: 317.1.

[0204]

Chemical Structure

[0205] Step 2. A microwave vial containing 2-(3-chloropyrazin-2-yl)propanedinitrile (1.00 g, 5.60 mmol), 3-methoxy-2,6-dimethyl-aniline (2.54 g, 16.8 mmol), and NMP (10 mL) was capped and stirred at 150 °C for 1 h, followed by stirring at 200 °C for 8 h. The reaction mixture was cooled to room temperature, poured into saturated aqueous NaHCO3, and diluted with water and EtOAc. The mixture was filtered through a pad of Celite, and the layers were separated. The organic layer was dried over Na2SO4, filtered, adsorbed onto silica, and purified by silica gel chromatography eluting with a gradient of 0-100% EtOAc in hexanes. The appropriate fractions were combined and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0-20% MeOH in DCM. The appropriate fractions were combined, concentrated and dried under vacuum to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (346 mg, 21% yield) as a pale yellow-brown solid.

[0206] Step 3. To a solution of 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (600 mg, 2.05 mmol) in DMF (10 mL) was added NBS (436 mg, 2.45 mmol). The mixture was stirred for 10 minutes, diluted with water, stirred for 20 minutes, and then filtered. The solid was washed with water and dried under vacuum. Purification by silica gel chromatography eluting with a gradient of 0 to 100% EtOAc in hexanes gave 6-amino-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (350 mg, 46% yield).

[0207] Step 4. To a solution of 6-amino-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (350 mg, 940 μmol) in DCM (10 mL) was added H2SO4 (1.88 mmol, 1 mL). The mixture was stirred for 60 min, quenched with crushed ice, and extracted with DCM. The organic phase was dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified on silica gel using a gradient of 0-20% MeOH in DCM to give 6-amino-2-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (300 mg, 82% yield).

[0208] Step 5. To a solution of 6-amino-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (300 mg, 769 μmol) in DCM (3 mL) was added a DCM solution of BBr3 (1 M, 2.31 mL). The mixture was stirred for 2 hours. The volatiles were removed in vacuo to give a crude mixture of 6-amino-3-bromo-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (263 mg, 91% yield), which was used in the next step without further purification. 1H NMR (400MHz, DMSO-d6) δ8.29(s,1H),7.55(s,2H),7.31(s,1H),7.21(s,1H),7.13-7.06(m,1H),6.96(d,J=8.3Hz,1H),1.78(s,3H),1.70(s,3H). MS:[M+1]:378.3.

[0209] [ka] Intermediate I (2-amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide) Step 1. To a solution of 2,3-dibromo-5-nitro-pyridine (20 g, 63.85 mmol) in NMP (120 mL) was added 2,6-dimethylpyridine (11.08 g, 103.4 mmol, 12 mL) and 3-methoxy-2,6-dimethyl-aniline (14 g, 95.23 mmol). The mixture was heated at 130 °C overnight. After cooling to room temperature, it was diluted with dropwise addition of water, stirred at room temperature for 20 minutes, and filtered. The solid was washed with water and dried under vacuum. The residue was purified using 2 × 330 g of silica gel eluting with a gradient of 10 to 30% EtOAc in heptane to give 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-5-nitro-pyridin-2-amine (12 g, 53% yield) as an off-white solid.

[0210] Step 2. To a solution of propanedinitrile (4.4 g, 66.6 mmol, 4.19 mL) in DME (120 mL) was added NaH (2.90 g, 66.9 mmol, 60% dispersion in mineral oil) portionwise over several times. The resulting mixture was stirred for 5 minutes, and then 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-5-nitro-pyridine-2-amine (11.6 g, 32.9 mmol) and PdCl2(dppf).CH2Cl2 (1.34 g, 1.65 mmol) were added. The mixture was stirred at 110 °C for 2 hours. The mixture was cooled to room temperature, diluted with water, and extracted twice with EtOAc. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 60% EtOAc in hexanes to give 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-5-nitro-pyrrolo[2,3-b]pyridine-3-carbonitrile (11 g, 99% yield) as a yellow solid.

[0211] Step 3. To a solution of 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-5-nitro-pyrrolo[2,3-b]pyridine-3-carbonitrile (1.130 g, 3.35 mmol) in THF (15 mL) were added Et3N (3.37 mmol, 470 μL), DMAP (45 mg, 368 μmol), and tert-butyl tert-butoxycarbonyl carbonate (1.47 g, 6.73 mmol). The mixture was stirred at 50 °C for 1 hour and then cooled to room temperature. Ethylenediamine (500 μL) was added and the mixture was stirred for 2 hours. The resulting mixture was diluted with water and extracted with DCM (2×). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 20 - 60% EtOAc to give tert-butyl N-[3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-5-nitro-pyrrolo[2,3-b]pyridin-2-yl]carbamate (1.27 g, 87% yield).

[0212] Step 4. To a solution of tert-butyl N-[3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-5-nitro-pyrrolo[2,3-b]pyridin-2-yl]carbamate (2.94 g, 6.72 mmol) in DCM (30 mL) and MeOH (30 mL) was added palladium carbon (10% w / w, 400 mg, 376 μmol). The mixture was stirred under 1 atm of H2 for 3 h. The suspension was filtered through a pad of celite and concentrated in vacuo to afford tert-butyl N-[5-amino-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridin-2-yl]carbamate (2.7 g, 99% yield) as an off-white solid.

[0213] Step 5. To a solution of tert-butyl N-[5-amino-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridin-2-yl]carbamate (15.1 g, 37.1 mmol) in a mixture of DMF (60 mL) and acetonitrile (80 mL) was added tert-butyl nitrite (5.72 g, 55.5 mmol, 6.6 mL), followed by copper(II) bromide (10 g, 44.8 mmol). The mixture was stirred at 60 °C for 20 min, then diluted with water, treated with ammonia, and extracted with EtOAc (3×). The combined organic extracts were washed with brine, dried over Na2SO4, and filtered. The filtrate was concentrated to dryness. The residue was purified by elution with a gradient of 0 - 5% EtOAc in DCM using a 3×330 silica gel column to afford tert-butyl N-[5-bromo-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridin-2-yl]carbamate (9.67 g, 55% yield) as an off-white solid. MS: 471.2 (M+H) + . The following by-product was also isolated by purification: tert-butyl (3-cyano-1-(3-methoxy-2,6-dimethylphenyl)-1H-pyrrolo[2,3-b]pyridin-2-yl)carbamate (350 mg, 2% yield).

[0214] Step 6. To a solution of tert-butyl N-[5-bromo-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridin-2-yl]carbamate (1.05 g, 2.23 mmol) in EtOH (15 mL) was added aqueous HCl solution (6 M, 6 mL) at 80 °C. The mixture was stirred for 20 minutes, then concentrated to dryness, co-evaporated with MeOH, treated with Et3N, and then concentrated to dryness. The residue was purified by reverse-phase flash chromatography on a C18 cartridge eluting with CH3CN / water / 0.1% formic acid to give 2-amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carbonitrile (515 mg, 60% yield) as an off-white solid.

[0215] Step 7. To a solution containing 2-amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carbonitrile (580 mg, 1.56 mmol) in a mixture of EtOH (6 mL) and water (2 mL) were added LiOH·H2O (500 mg, 11.9 mmol) and H2O2 (27% w / w aqueous solution, 21.02 mmol, 650 μL). The mixture was stirred at 60 °C for 20 minutes, cooled to room temperature, diluted with water, and filtered. The solid was washed with water and dried under vacuum to give 2-amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (600 mg, 99% yield) as an off-white solid. 1 1H NMR (400 MHz, DMSO-d6) δ 8.21 (d, J = 2.0 Hz, 1H), 7.77 (d, J = 2.0 Hz, 1H), 7.20 (dt, J = 8.4, 0.7 Hz, 1H), 7.13 (s, 2H), 7.05 (d, J = 8.5 Hz, 1H), 6.83 (s, 2H), 3.73 (s, 3H), 1.75 (d, J = 0.7 Hz, 3H), 1.65 (s, 3H). MS: [M+1]: 469.1.

[0216]

Chemical Structure

[0217] Step 2. A solution of tert-butyl 2-((6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethylphenyl)-5H-pyrrolo[2,3-b]pyrazine-2-l)ethynyl)pyrrolidine-1-carboxylate (0.13 g, 0.55 mmol) in methanol was added to palladium on carbon (10% w / w, 50% water). The suspension was stirred under a hydrogen atmosphere for 2 hours. The reaction mixture was filtered through Celite and washed with methanol. The filtrate was concentrated in vacuo, and the residue was purified by silica gel chromatography eluting with 30% EtOAc in hexanes to give tert-butyl 2-(2-(6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethylphenyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl)ethyl)pyrrolidine-1-carboxylate (0.105 g, 49% yield) as an off-white solid.

[0218] Step 3. The same procedure used for O-Me deprotection using BBr3 as used for compound 35 gave a residue which was purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethylphenyl)-2-(2-(pyrrolidin-2-yl)ethyl)-5H-pyrrolo[2,3-b]pyrazine-7-carboxamide (2.7 mg, 3.5% yield) as a white solid. 1 H NMR(400MHz,DMSO-d6)δ9.79(bs,1H),8.40(s,1H),7.65(s,1H),7.47(s,1H),7.35(s,2H),7.27(s,1H),7.07(d,J=8Hz,1H),6.95(d, J=8.4Hz,1H),3.39(s,2H),3.06(s,1H),2.99(s,1H),2.80(s,2H),2.02(s,3H),1.83(s,1H),1.76(s,3H),1.68(s,3H),1.46(s,1H). MS:[M+1]:395.5.

[0219] [ka] Compound 6 (2-amino-5-(cyclopentan-1-yl)-1-(5-hydroxy-2-methyl-phenyl)pyrrolo[3,2-b]pyridine-3-carboxamide) To a solution of intermediate G (33 mg, 104 μmol) in dioxane (1.5 mL) were added 2-(cyclopentan-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (40 mg, 206 μmol), PdCl(dppf).CHCl (8 mg, 10 μmol), and aqueous NaCO (2 M, 200 μL). The mixture was stirred at 100 °C for 5 h. The volatiles were removed in vacuo. The residue was purified by preparative HPLC to give 2-amino-5-(cyclopentan-1-yl)-1-(5-hydroxy-2-methyl-phenyl)pyrrolo[3,2-b]pyridine-3-carboxamide (5 mg, 14% yield) as an off-white solid. 1H NMR(400MHz,DMSO-d6)δ9.77(s,1H),8.03(s,1H),7.25(d,J=8.3Hz,1H),6.99(d,J=8.1Hz,2H),6.91(s,2H),6.88(dd,J=8.3,2.6 Hz,1H),6.82(d,J=8.1Hz,1H),6.66(d,J=2.5Hz,1H),6.41(t,J=2.1Hz,1H),2.72(m,2H),2.50(m,2H),1.94(m,2H),1.78(s,3H). MS:[M+1]:349.1.

[0220] [ka] Compound 16 (2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-(3-morpholinoprop-1-ynyl)pyrrolo[3,2-b]pyridine-3-carboxamide) A solution of 4-prop-2-ynylmorpholine (40 mg, 0.32 mmol), Intermediate G (50 mg, 0.16 mmol), copper(I) iodide (3 mg, 16 μmol), NaCO (70 mg, 0.66 mmol), tri-tert-butylphosphonium tetrafluoroborate (9 mg, 31 μmol), and PdCl (3 mg, 17 μmol) in DMF (2 mL) was degassed under vacuum and backfilled with nitrogen. The mixture was stirred at 100 °C for 5 h. The mixture was purified by preparative HPLC eluting with CHCN / water / 10 mM ammonium bicarbonate (pH 10). The desired fractions were combined and lyophilized to give 2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-(3-morpholinoprop-1-ynyl)pyrrolo[3,2-b]pyridine-3-carboxamide (22 mg, 35% yield) as an off-white solid. 11H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 7.74 (d, J = 3.6 Hz, 1H), 7.38 - 7.16 (m, 1H), 7.06 (s, 3H), 6.98 (d, J = 8.1 Hz, 1H), 6.88 (dd, J = 8.4, 2.6 Hz, 1H), 6.83 (d, J = 8.0 Hz, 1H), 6.66 (d, J = 2.5 Hz, 1H), 3.57 (m, 4H), 3.50 (s, 2H), 2.52 - 2.47 (m, 4H), 1.77 (s, 3H). MS: [M+1]: 406.2.

[0221]

Chem.

[0222]

Chem.

[0223] [ka] Compound 27 (2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-(1,2,3,6-tetrahydropyridin-5-yl)pyrrolo[3,2-b]pyridine-3-carboxamide HCl salt) To a solution of tert-butyl 5-[2-amino-3-carbamoyl-1-(5-hydroxy-2-methyl-phenyl)pyrrolo[3,2-b]pyridin-5-yl]-3,6-dihydro-2H-pyridine-1-carboxylate (75 mg, 0.162 mmol, prepared as in Compound 6) in MeOH (1 mL) was added dioxane containing HCl (4 M, 0.5 mL). The mixture was stirred for 2 hours. Volatiles were removed under vacuum. The residue was dissolved in water and CH3CN and then lyophilized to give 2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-(1,2,3,6-tetrahydropyridin-5-yl)pyrrolo[3,2-b]pyridine-3-carboxamide HCl salt (64 mg, 99% yield) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 9.20 (s, 2H), 8.01 - 7.41 (m, 1H), 7.26 (d, J = 8.4, Hz, 1H), 7.12 (d, J = 8.3 Hz, 1H), 7.02 (s, 3H), 6.92 - 6.82 (m, 2H), 6.72 - 6.61 (m, 2H), 4.11 (s, 2H), 3.44 (m 2H), 3.19 (m, 2H), 1.77 (s, 3H). MS: [M+1]: 464.2.

[0224]

Chemical Structure

[0225] Step 2. For OMe deprotection using BBr3, the same procedure as used for Compound 35 was carried out on the appropriate intermediate (23 mg, 58 μmol) to give a residue which was purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (10 mg, 45% yield) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.63 (s, 1H), 8.49 (s, 1H), 7.91 (d, J = 3.2 Hz, 1H), 7.78 (d, J = 3.2 Hz, 1H), 7.61 (s, 2H), 7.40 (s, 1H), 7.29 (s, 1H), 7.06 (d, J = 8.4 Hz, 1H), 6.92 (d, J = 8.3 Hz, 1H), 1.77 (s, 3H), 1.69 (s, 3H). MS: [M+1]: 381.2.

[0226]

Chem.

[0227] Step 2. To a solution containing 6-amino-2-benzyloxy-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (650 mg, 1.36 mmol) in dioxane (10 mL) and water (3 mL) were added Pd(PPh3)4 (80 mg, 69 μmol) and K2CO3 (800 mg, 5.79 mmol). The mixture was degassed under vacuum and then refilled with nitrogen three times. Then, 2,4,6-trimethyl-1,3,5,2,4,6-trioxatriborinane (712 mg, 2.84 mmol, 0.8 mL) was added and the final mixture was stirred at 100 °C for 18 hours. The mixture was cooled to room temperature, diluted with EtOAc, washed with water and brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 45% EtOAc in hexane to give 6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-methyl-pyrrolo[2,3-b]pyrazine-7-carbonitrile (300 mg, 53% yield) as an off-white solid.

[0228] Step 3. For nitrile hydrolysis using sulfuric acid, the same procedure used for compound 164 was carried out on the appropriate intermediate (260 mg, 0.629 mmol) to give 6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-methyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (205 mg, 96% yield) as an off-white solid.

[0229] Step 4. To a solution of 6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-methyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (206 mg, 0.603 mmol) and CsCO (390 mg, 1.20 mmol) in DMF (2 mL) was added 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (235 mg, 0.658 mmol). The mixture was stirred for 1 h, then diluted with water and extracted with EtOAc (4x). The combined organic extracts were washed with brine, dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0 to 70% EtOAc in hexanes to give [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-methyl-pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (160 mg, 56% yield) as an off-white solid.

[0230] Step 5. A mixture of tributyl(thiazol-2-yl)stannane (83 mg, 0.223 mmol, 70 μL), trifluoromethanesulfonate [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-methyl-pyrrolo[2,3-b]pyrazin-2-yl] (50 mg, 0.106 mmol), CuI (3 mg, 16 μmol), LiCl (7 mg, 0.165 mmol), and PdCl(dppf).CHCl (8 mg, 11 μmol) in DMF (1.5 mL) was degassed under vacuum and then backfilled with nitrogen. The reaction mixture was stirred at 110 °C for 4 h, cooled to room temperature, and concentrated under vacuum. The residue was purified by preparative HPLC to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-3-methyl-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (22 mg, 51% yield) as an off-white solid.

[0231] Step 6. For OMe deprotection using BBr3, the same procedure used for compound 35 was carried out on the appropriate intermediate (22 mg, 53 μmol) to give a residue that was purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-methyl-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (5 mg, 24% yield) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ9.64(s,1H),7.94(d,J=3.3Hz,1H),7.76(d,J=3.3Hz,1H),7.47(s,2H),7.35(s ,1H),7.22(s,1H),7.06(d,J=8.3Hz,1H),6.92(d,J=8.3Hz,1H),2.76(s,3H),1.77(s,3H),1.69(s,3H). MS:[M+1]:395.2.

[0232] [ka] Compound 33 (6-amino-3-bromo-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (compound 28, 60 mg, 0.15 mmol) in DMF (1 mL) was added NBS (30 mg, 0.17 mmol). The mixture was stirred for 18 h, diluted with water, treated with 20% aqueous NaSO, and extracted with EtOAc (3x). The combined organic extracts were washed with water, brine, dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 20 to 100% EtOAc in hexanes to give 6-amino-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (45 mg, 63% yield) as an off-white solid.

[0233] Step 2. For OMe deprotection using BBr, the same procedure used for compound 35 was performed on the appropriate intermediate (35 mg, 74 μmol) to give a residue that was purified by preparative HPLC to give 6-amino-3-bromo-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (10 mg, 29% yield) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ9.71(s,1H),7.97(d,J=3.3Hz,1H),7.88(d,J=3.3Hz,1H),7.75( s,2H),7.45(s,1H),7.19-7.00(m,2H),6.94(d,J=8.3Hz,1H),1.79(s,3H),1.71(s,3H). MS:[M+1]:460.2.

[0234] [ka] Compound 35 (6-Amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[2-(trifluoromethyl)-4-pyridyl]pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of Intermediate D (50 mg, 0.109 mmol) in dioxane (1 mL) were added PdCl2(dppf).CH2Cl2 (8 mg, 10 μmol), [2-(trifluoromethyl)-4-pyridyl]boronic acid (40 mg, 0.209 mmol), and aqueous Na2CO3 solution (2 M, 200 μL). The mixture was degassed under vacuum and refilled with nitrogen. The reaction mixture was stirred at 100 °C for 4 hours, cooled to room temperature, diluted with water, and then filtered. The solid was washed with water, dried under vacuum, and finally purified by silica gel chromatography eluting with a gradient of 20 - 100% EtOAc in hexane to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[2-(trifluoromethyl)-4-pyridyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (36 mg, 72% yield) as an off-white solid.

[0235] Step 2. (General procedure for OMe deprotection using BBr3) To a solution of 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[2-(trifluoromethyl)-4-pyridyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (36 mg, 79 μmol) in DCM (1 mL) was added BBr3 (1 M in DCM, 230 μL). The mixture was stirred for 1 hour. The volatile substances were removed under vacuum. The residue was dissolved in MeOH and concentrated to dryness again. Then it was dissolved in MeOH, Et3N (100 μL) was added, and the mixture was concentrated to dryness again. The residue was purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[2-(trifluoromethyl)-4-pyridyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (16 mg, 46% yield) as an off-white solid. 11H NMR (400 MHz, DMSO-d6) δ 9.63 (s, 1H), 8.81 (m, 1H), 8.62 (s, 1H), 8.48 (s, 1H), 8.41 (m, 1H), 7.63 (s, 2H), 7.45 (s, 1H), 7.33 (s, 1H), 7.07 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 1.77 (s, 3H), 1.69 (s, 3H). MS: [M+1]: 443.2.

[0236] [Chemical] Compound 46 (6-Amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[4-(methylcarbamoyl)-1-piperidyl]pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of Intermediate D (50 mg, 0.109 mmol) in DMSO (1 mL) was added N-methylpiperidine-4-carboxamide (80 mg, 0.563 mmol). The mixture was stirred at 130 °C for 2 h in a sealed vial, then cooled to room temperature and purified by preparative HPLC to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[4-(methylcarbamoyl)-1-piperidyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (14 mg, 28% yield) as an off-white solid.

[0237] Step 2. For the OMe deprotection using BBr3, the same procedure as used for Compound 35 was carried out on the appropriate intermediate (15 mg, 32 μmol) to give a residue, which was purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[4-(methylcarbamoyl)-1-piperidyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (10 mg, 69% yield) as an off-white solid. 11H NMR (400 MHz, DMSO-d6) δ 9.51 (s, 1H), 7.72 (d, J = 4.8 Hz, 1H), 7.35 (s, 1H), 7.27 (s, 1H), 7.14 - 6.96 (m, 4H), 6.87 (d, J = 8.2 Hz, 1H), 4.13 (d, J = 12.4 Hz, 2H), 2.85 - 2.68 (m, 2H), 2.53 (d, J = 4.6 Hz, 3H), 2.35 - 2.20 (m, 1H), 1.74 (m, 5H), 1.65 (s, 3H), 1.64 - 1.50 (m, 2H). MS: [M+1]: 438.2.

[0238] [Chemical formula] Compound 97 (1-[6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-pyrazin-2-yl-pyrrolo[2,3-b]pyrazin-7-yl]ethenone) Step 1. To a solution of Intermediate C (2.5 g, 6.26 mmol) in THF (20 mL) was added a THF solution of MeMgBr (3 M, 6.50 mL) at 0 °C. The mixture was warmed and stirred for 18 hours. An additional amount of THF solution of MeMgBr (3 M, 4.00 mL) was added and the mixture was stirred for an additional 5 hours. The resulting mixture was quenched with saturated aqueous NH4Cl, diluted with water, and extracted with EtOAc. The organic extract was washed with water and brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by elution with a gradient of 0 - 30% EtOAc in hexane through a silica gel column to give 1-[6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-7-yl]ethanone (120 mg, 5% yield).

[0239] Step 2. To a solution of [[6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-7-yl]ethanone (100 mg, 0.240 mmol) in DCM (1 mL) was added TFA (500 μL). The mixture was stirred at 50 °C for 10 h. The volatiles were removed under vacuum. The residue was purified by preparative HPLC to give 1-[6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-7-yl]ethanone (43 mg, 55% yield).

[0240] Step 3. To a mixture containing 1-[6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-7-yl]ethanone (43 mg, 0.132 mmol) and Cs2CO3 (50 mg, 0.153 mmol) in DMF (1 mL) was added 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (52 mg, 0.146 mmol). The mixture was stirred for 1 h. The volatiles were removed under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 70% EtOAc in hexanes to give [7-acetyl-6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (46 mg, 76% yield) as an off-white solid.

[0241] Step 4. To a solution of trifluoromethanesulfonic acid [7-acetyl-6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] (46 mg, 0.100 mmol) in DMF (1 mL) were added LiCl (9 mg, 0.212 mmol), tributyl(pyrazin-2-yl)stannane (74 mg, 0.200 mmol) and PdCl2(dppf).CH2Cl2 (7 mg, 9.6 μmol). The mixture was stirred at 120 °C for 10 h. Volatiles were removed under vacuum. The residue was purified by preparative HPLC to give 1-[6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-pyrazin-2-yl-pyrrolo[2,3-b]pyrazin-7-yl]ethanone (38 mg, 97% yield) as an off-white solid.

[0242] Step 5. For the OMe deprotection using BBr3, the same procedure as used for compound 35 was carried out on the appropriate intermediate (38 mg, 98 μmol) to give a residue which was purified by preparative HPLC to give 1-[6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-pyrazin-2-yl-pyrrolo[2,3-b]pyrazin-7-yl]ethanone (18 mg, 49% yield) as an off-white solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 9.55 (s, 1H), 8.72 (s, 1H), 8.69 - 8.62 (m, 2H), 8.13 (s, 2H), 7.07 (d, J = 8.3 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 2.76 (s, 3H), 1.77 (s, 3H), 1.69 (s, 3H). MS: [M+1]: 375.

[0243]

Chemical Structure

[0244] Step 2. (1,10-Phenanthroline)(trifluoromethyl)copper(I) (900 mg, 2.88 mmol) was added to a solution of 6-amino-2-benzyloxy-3-iodo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (745 mg, 1.42 mmol) in DMF (10 mL). The mixture was stirred at 70 °C for 4 hours. The volatile substances were removed under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 20 - 60% EtOAc in hexane to give 6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (300 mg, 45% yield).

[0245] Step 3. A solution of 6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (300 mg, 0.642 mmol) in sulfuric acid (1 mL) was stirred for 5 hours, poured onto crushed ice, neutralized with ammonia solution, and the resulting precipitate was filtered. The precipitate was washed with water and dried under vacuum to give 6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (232 mg, 91% yield) as a yellow solid.

[0246] Step 4. To a solution of 4.6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (232 mg, 0.587 mmol) and Cs2CO3 (200 mg, 0.615 mmol) in DMF (2 mL) was added 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (210 mg, 0.588 mmol). The mixture was stirred for 1 h, diluted with water, and stirred for 20 min. The resulting precipitate was filtered, washed with water, and dried under vacuum. Further purification by silica gel chromatography eluting with a gradient of 20 - 100% EtOAc in hexanes gave trifluoromethanesulfonic acid [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazin-2-yl] (190 mg, 61% yield) as an off-white solid.

[0247] Step 5. To a solution of trifluoromethanesulfonic acid [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazin-2-yl] (90 mg, 0.171 mmol) in dioxane (1 mL) were added PdCl2(dppf).CH2Cl2 (14 mg, 17 μmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (30 mg, 0.195 mmol), and aqueous Na2CO3 (2 M, 100 μL). The mixture was stirred at 120 °C for 18 h. Volatiles were removed under vacuum. The residue was purified by preparative HPLC to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)-2-vinyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (10 mg, 14% yield) as an off-white solid.

[0248] Step 6. For OMe deprotection using BBr, the same procedure used for compound 35 was carried out on the appropriate intermediate (10 mg, 25 μmol) to give a residue that was purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)-2-vinyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (3 mg, 31% yield) as an off-white solid. 1 H NMR (400MHz, DMSO-d6) δ9.67(s,1H),7.85(s,2H),7.41(m,2H),7.10-6.89(m,3H),6.50(m,1H),5.60(m,1H),1.76(s,3H),1.68(s,3H). MS:[M+1]:392.2.

[0249] [ka] Compound 110 (6-amino-2-cyclopropyl-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Procedure 1. To a suspension of magnesium turnings (140 mg, 5.76 mmol) in THF (10 mL) was added iodine (13 mg, 52 μmol). The mixture was stirred for 10 minutes, then CD3I (5.14 mmol, 320 μL) was added and the mixture was stirred under nitrogen for 18 hours to form an off-white suspension. ZnCl2 in THF (0.5 M, 10.5 mL) was added dropwise to the mixture. After the addition, the mixture was stirred for 20 minutes, then 6-amino-2-benzyloxy-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (500 mg, 1.05 mmol) and Pd(PPh3)4 (120 mg, 0.103 mmol) were added. The final mixture was stirred at 70 °C for 6 hours. The reaction was quenched with 1 M HCl, diluted with water, and extracted with EtOAc (2×). The combined organic extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 60% EtOAc in hexanes to give 6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (328 mg, 75% yield) as an off-white solid.

[0250] Procedure 2. A mixture containing 6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (328 mg, 0.788 mmol) in H2SO4 (2 mL) was stirred for 4 hours. The mixture was cooled to 0 °C and then neutralized to pH 7 using concentrated aqueous ammonia. The resulting mixture was lyophilized, the residue was triturated with water, and filtered. The solid was dried in vacuo to give 6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (220 mg, 81% yield) as an off-white solid.

[0251] Step 3. To a solution of 6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (232 mg, 0.674 mmol) in DMF (3 mL) was added CsCO (320 mg, 0.982 mmol) and 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (360 mg, 1.01 mmol). The mixture was stirred for 1 h. The volatiles were removed in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 20 to 100% EtOAc in hexanes to give [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (200 mg, 62% yield) as an off-white solid.

[0252] Step 4. To a solution of [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (200 mg, 0.420 μmol) in DMF (3 mL) was added lithium chloride (36 mg, 0.849 mmol) and tributyl(cyclopropyl)stannane (275 mg, 0.831 mmol). The mixture was stirred at 120 °C for 10 hours. The volatiles were removed in vacuo. The residue was purified by preparative HPLC to give 6-amino-2-cyclopropyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (80 mg, 52% yield) as an off-white solid.

[0253] Step 5. For OMe deprotection using BBr, the same procedure used for compound 35 was carried out on the appropriate intermediate (35 mg, 95 μmol) to give a residue that was purified by preparative HPLC to give 6-amino-2-cyclopropyl-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (20 mg, 59% yield) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ9.49(s,1H),7.24(s,1H),7.14-6.98(m,4H),6.89(d, J=8.2Hz,1H),2.11(m,1H),1.84-1.68(s,3H),1.64(s,3H),1.04-0.81(m,4H). MS:[M+1]:356.2.

[0254] Chiral SFC separation of compound 110 (20 mg, 0.056 mmol) (apparatus: Waters Prep 15 SFC-MS; column: Phenomenex Lux Cellulose-2, 10 × 250 mm, 5 μm; conditions: isocratic with 45% CO and 55% MeOH; flow rate: 10 mL / min) gave compounds 111 and 112. [ka] Compound 111 from chiral SFC separation of compound 110. Peak 1 (retention time 5.33 min, 99.95%): S-6-amino-2-cyclopropyl-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (7.8 mg) was obtained as an off-white solid. 1H NMR (400 MHz, DMSO-d6) δ 9.49 (s, 1H), 7.24 (s, 1H), 7.14-6.98 (m, 4H), 6.89 (d, J = 8.2 Hz, 1H), 2.11 (m, 1H), 1.84-1.68 (s, 3H), 1.64 (s, 3H), 1.04-0.81 (m, 4H). MS: [M+1]: 356.2. [ka] Compound 112 from chiral SFC separation of Compound 110. Peak 2 (retention time 6.00 min, 99.78%): R-6-Amino-2-cyclopropyl-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (5.3 mg) was obtained as an off-white solid. 1H NMR (400 MHz, DMSO-d6) δ 9.49 (s, 1H), 7.24 (s, 1H), 7.14 - 6.98 (m, 4H), 6.89 (d, J = 8.2 Hz, 1H), 2.11 (m, 1H), 1.84 - 1.68 (s, 3H), 1.64 (s, 3H), 1.04 - 0.81 (m, 4H). MS: [M + 1]: 356.2.

[0255]

Chemical formula

[0256] Step 2. To a solution of 2-amino-5-cyclopropyl-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carbonitrile (73 mg, 0.220 mmol) in EtOH (1.5 mL) and water (300 μL) was added LiOH.HO (50 mg, 1.19 mmol) and HO (700 μL, 27% w / w aqueous solution). The mixture was stirred at 60 °C for 20 min, after which the volatiles were removed in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 20–60% EtOAc in hexanes to give 2-amino-5-cyclopropyl-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (22 mg, 29% yield) as an off-white solid.

[0257] Step 3. The same procedure used for compound 35 for OMe deprotection using BBr was carried out on the appropriate intermediate (22 mg, 63 μmol) to give a residue that was purified by preparative HPLC to give 2-amino-5-cyclopropyl-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (15 mg, 71% yield) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ9.43(s,1H),7.58(s,1H),7.50(s,1H),7.00(d,J=8.3Hz,1H),6.86(m,3H),6.69(s,2H),1.88 m,1H),1.69(s,3H),1.61(s,3H),0.86(m,2H),0.81-0.68(m,2H). MS:[M+1]:337.2.

[0258] Chiral SFC separation of compound 116 (410 mg, 1.22 mmol) (apparatus: Waters Prep 100 SFC-MS; column: Phenomenex Lux Cellulose-2, 30 × 250 mm, 5 μm; conditions: isocratic with 1:1 50% ACN / EtOH at 50% CO; flow rate: 70 mL / min) gave compounds 117 and 118. [ka] Compound 117 from chiral SFC separation of Compound 116. Peak 1 (retention time 5.60 min, 99.83%): S-2-Amino-5-cyclopropyl-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (130 mg) was obtained as an off-white solid. 1H NMR (400 MHz, DMSO-d6) δ 9.52 (s, 1H), 8.12 (d, J = 2.2 Hz, 1H), 8.08 - 7.92 (m, 2H), 7.46 (s, 1H), 7.00 (d, J = 8.2 Hz, 2H), 6.86 (d, J = 8.3 Hz, 1H), 2.03 (m, 1H), 1.70 (s, 3H), 1.59 (s, 3H), 0.96 (m, 2H), 0.68 (m, 2H). MS: [M+1]: 337.2.

Chem.

[0259]

Chem.

[0260] Step 2. To a suspension of NaH (1.08 g, 24.9 mmol, 60% dispersion in mineral oil) in DME (60 mL) was added dropwise a solution of propanedinitrile (1.62 g, 24.6 mmol) in DME (15 mL). After stirring for 30 minutes, a solution of 3-bromo-5-chloro-N-(3-methoxy-2,6-dimethyl-phenyl)pyridin-2-amine (4.00 g, 11.7 mmol) in DME (15 mL) and PdCl2(dppf).CH2Cl2 (1.08 g, 1.32 mmol) were added. Nitrogen was bubbled through the reaction mixture and then the mixture was stirred at 100 °C for 5 hours. The reaction mixture was cooled to room temperature and ice water (250 mL) was added dropwise. The resulting precipitate was collected by filtration and washed with water. The solid was air dried and then co-evaporated with toluene (2×) and dried under vacuum to give 4.67 g of a crude product. Purification by silica gel chromatography (dry loading) eluting with a gradient of 0 - 100% EtOAc in heptane. The fractions were combined, concentrated, and dried under vacuum to give 2-amino-5-chloro-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carbonitrile (3.27 g, 85% yield) as an off-white crystalline solid.

[0261] Step 3. To a suspension of 3,2-amino-5-chloro-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carbonitrile (7.50 g, 23.0 mmol) in water (60 mL) and reagent alcohol (180 mL), 98% LiOH.H2O (7.22 g, 172 mmol) and H2O2 (27% w / w aqueous solution, 9.8 mL) were added. The mixture was stirred at 60 °C for 30 minutes and then cooled to room temperature. Water was added dropwise (500 mL), the solid was collected by filtration, washed with water and air-dried. The filtrate was diluted with water to obtain a second collected solid. Finally, the filtrate was extracted with EtOAc (3×). The combined organic extracts were dried over Na2SO4, filtered, concentrated and then dried under vacuum to obtain a third collected crude product. The combined crude materials were purified by silica gel chromatography using a gradient of 50 - 100% EtOAc in heptane. The pure fractions were combined, concentrated and dried under vacuum to obtain 2-amino-5-chloro-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (3.90 g, 49% yield) as a pale yellow solid. Alternatively, nitrile hydrolysis could be carried out under H2SO4 conditions (using the same procedure as used for Compound 164) to obtain 2-amino-5-chloro-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide in quantitative yield.

[0262] Step 4. For OMe deprotection using BBr3, the same procedure as used for Compound 35 was used on the appropriate intermediate (3.90 g, 11.3 mmol) to obtain a residue which was co-evaporated with MeOH (4×), dried under vacuum, triturated with saturated aqueous NaHCO3 and filtered. The crude product was purified by silica gel chromatography on silica using a gradient of 0 - 20% MeOH in CH2Cl2 to obtain 2-amino-5-chloro-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (3.54 g, 95% yield) as a pale off-white solid. 1H NMR(400MHz,DMSO-d6)δ9.54(s,1H),8.14(d,J=2.2Hz,1H),7.74(d,J=2.1Hz,1H),7.14(br s,2H),7.06(d,J=8.3Hz,1H),6.91(d,J=8.3Hz,1H),6.86(br s,2H),1.74(s,3H),1.65(s,3H). MS:[M+1]:331.1.

[0263] Chiral SFC separation of compound 132 (3.54 g, 10.7 mmol) (apparatus: Waters Prep 100 SFC-MS; column: Phenomenex Lux Cellulose-2, 30 × 250 mm, 5 μm; conditions: isocratic with 1:1 45% ACN / EtOH at 55% CO; flow rate: 70 mL / min) gave compounds 133 and 134. [ka] Compound 133 from chiral SFC separation of compound 132. Peak 1 (retention time 5.37 min, 99.70%): S-2-amino-5-chloro-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (1.26 g) was obtained as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ9.54(s,1H),8.14(d,J=2.2Hz,1H),7.74(d,J=2.1Hz,1H),7.14(br s,2H),7.06(dt,J=8.2,0.7Hz,1H),6.91(d,J=8.3Hz,1H),6.86(br s,2H),1.74(d,J=0.7Hz,3H),1.65(s,3H). MS:[M+1]:331.1. [ka] Compound 134 from chiral SFC separation of compound 132. Peak 2 (retention time 7.79 min, 99.19%): R-2-amino-5-chloro-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (1.26 g).1 1H NMR (400 MHz, DMSO-d6) δ 9.54 (s, 1H), 8.14 (d, J = 2.2 Hz, 1H), 7.74 (d, J = 2.1 Hz, 1H), 7.14 (br s, 2H), 7.06 (dt, J = 8.2, 0.7 Hz, 1H), 6.91 (d, J = 8.3 Hz, 1H), 6.86 (br s, 2H), 1.74 (d, J = 0.7 Hz, 3H), 1.65 (s, 3H). MS: [M+1]: 331.1.

[0264]

Chem.

[0265] Step 2. For the nitrile hydrolysis using sulfuric acid, the same procedure as used for Compound 164 was carried out on the appropriate intermediate (28 mg, 0.079 mmol) to give 6-amino-5-(5-methoxy-2-methyl-phenyl)-3-phenyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (20 mg, 68% yield) as a pale yellow solid.

[0266] Step 3. For the OMe deprotection using BBr3, the residue was obtained by the same procedure as that used for Compound 35, and purified by preparative HPLC to give 6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-phenyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (11 mg, 57% yield) as a white cottony solid. 1 1H NMR (400 MHz, DMSO-d6) δ 9.71 (br s, 1H), 8.73 (s, 1H), 7.92 - 7.76 (m, 2H), 7.49 (br s, 2H), 7.45 - 7.37 (m, 3H), 7.35 - 7.28 (m, 2H), 7.26 (br s, 1H), 6.92 (dd, J = 8.3, 2.6 Hz, 1H), 6.77 (d, J = 2.5 Hz, 1H), 1.89 (s, 3H). MS: [M + 1]: 360.2.

[0267]

Chem.

[0268] Procedure 2. To a solution of 6 - amino - 5 - [5 - (methoxymethoxy)-2 - methyl - phenyl]-3-(3 - pyridylmethoxy)pyrrolo[2,3 - b]pyrazine - 7 - carbonitrile (65.0 mg, 0.156 mmol) in MeOH (1.5 mL) was added dioxane containing HCl (4 M, 1.50 mL). After stirring for 75 min, the reaction was concentrated and dried under vacuum. The crude 6 - amino - 5 - (5 - hydroxy - 2 - methyl - phenyl)-3-(3 - pyridylmethoxy)pyrrolo[2,3 - b]pyrazine - 7 - carbonitrile, assumed to be the bis HCl salt, was carried on to the next step without further purification.

[0269] Step 3. To a solution of crude 6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-(3-pyridylmethoxy)pyrrolo[2,3-b]pyrazine-7-carbonitrile (70 mg, 0.156 mmol assuming the bis-HCl salt) in MeOH (1.0 mL) was added aqueous NaOH (4 M, 1.0 mL). The reaction mixture was transferred to a preheated 90 °C heat block and stirred for 18 h. After cooling to room temperature, the mixture was neutralized with 3 N HCl and diluted with water. The precipitate was collected by filtration, washed with water, and then air-dried. Purification by preparative HPLC afforded 6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-(3-pyridylmethoxy)pyrrolo[2,3-b]pyrazine-7-carboxamide (4 mg, 7% yield) as a white fluffy solid. 1 H NMR(400MHz,DMSO-d6)δ9.68(br s,1H),8.48(d,J=2.2Hz,1H),8.42(dd,J=4.8,1.7Hz,1H),7.85(s,1H),7.65(dt,J =7.8,2.0Hz,1H),7.26(ddd,J=7.9,4.9,0.9Hz,1H),7.23(d,J=8.6Hz,1H),7.15(br s,1H),7.04(br s,1H),7.00(br s,2H),6.87(dd,J=8.3,2.6Hz,1H),6.66(d,J=2.5Hz,1H),5.09(d,J=12.2Hz,1H),5.08(d,J=12.2Hz,1H),1.72(s,3H). MS:[M+1]:391.2.

[0270] [ka] Compound 160 (6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-[2-(3-pyridyl)ethynyl]pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. Intermediate F (251 mg, 0.549 mmol), Pd(PPh3)4 (65 mg, 0.056 mmol), and CuI (43 mg, 0.023 mmol) were charged into a microwave vial through which nitrogen was flowing, and DMF (2.5 mL) containing 3-ethynylpyridine (72 mg, 0.698 mmol) was added, followed by Et3N (610 μL, 4.39 mmol). The vial was capped and then transferred to a preheated heat block (120 °C). After 1 hour, the reaction mixture was concentrated under vacuum, then taken up in THF and adsorbed onto silica. The volatile materials were evaporated under vacuum, and the residue was purified by silica gel chromatography eluting with a gradient of 0 - 100% EtOAc in hexanes followed by a gradient of 0 - 20% MeOH in EtOAc to give 6-amino-5-[5-(methoxymethoxy)-2-methyl-phenyl]-3-[2-(3-pyridyl)ethynyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile (260 mg, 99%) as a light brown solid.

[0271] Step 2. To a suspension of 6-amino-5-[5-(methoxymethoxy)-2-methyl-phenyl]-3-[2-(3-pyridyl)ethynyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile (225 mg, 0.548 mmol) in MeOH (3 mL) was added dioxane (4 M, 3 mL) containing HCl. After stirring for 30 minutes, the reaction mixture was concentrated to dryness and then dried under vacuum to give 6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-[2-(3-pyridyl)ethynyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile (319 mg, bis HCl salt) as a light brown viscous solid, which was used in the next step without further purification.

[0272] Step 3. 6-Amino-5-(5-hydroxy-2-methyl-phenyl)-3-[2-(3-pyridyl)ethynyl]pyrrolo[2,3-b]pyrazine-7-carbonitrile bis HCl salt (241 mg, 0.548 mmol) was stirred in concentrated H2SO4 (2 mL). After 3 days, the reaction mixture was quenched with crushed ice, placed in an ice bath, and made alkaline (pH ~10) with 1:1 NH4OH / H2O. The solid was collected by filtration and air-dried overnight to give the crude product as an off-yellow solid (249 mg). A portion (67 mg) was purified by preparative HPLC to give 6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-[2-(3-pyridyl)ethynyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (26 mg, 46% calculated yield) as a pale yellow fluffy solid. 1 1H NMR (400 MHz, DMSO-d6) δ 9.77 (br s, 1H), 8.75 (dd, J = 2.3, 0.9 Hz, 1H), 8.57 (dd, J = 4.9, 1.7 Hz, 1H), 8.42 (s, 1H), 7.98 (dt, J = 7.9, 1.9 Hz, 1H), 7.73 (br s, 2H), 7.44 (ddd, J = 8.0, 4.9, 0.9 Hz, 1H), 7.35 (br d, J = 4.4 Hz, 2H), 7.29 (d, J = 8.4 Hz, 1H), 6.94 (dd, J = 8.3, 2.6 Hz, 1H), 6.77 (d, J = 2.6 Hz, 1H), 1.85 (s, 3H). MS: [M+1]: 385.3.

[0273]

Chemical Structure

[0274]

Chemical Structure

[0275] Step 2. The reaction mixture was divided into three vials, each containing 1 / 3 of the volume. A microwave vial was charged with 2-(3-chloro-5,6-dimethyl-pyrazin-2-yl)propanedinitrile (3.04 g, 14.7 mmol), 3-methoxy-2,6-dimethyl-aniline (6.61 g, 43.7 mmol), potassium tert-butoxide (3.30 g, 29.4 mmol), and Pd-PEPPSI™-SIPr catalyst (513 mg, 0.752 mmol) and flushed with nitrogen three times. Then, anhydrous NMP (30 mL) was added, flushed with nitrogen again, capped, and subjected to microwave irradiation (at 100 °C) for 30 minutes. The vials were combined and diluted with EtOAc, saturated aqueous NH4Cl, and water. The layers were separated, and the aqueous layer was extracted twice with EtOAc. The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated in vacuo. The crude product was purified by silica gel chromatography eluting with a gradient of 0 to 100% EtOAc in heptane. The desired fractions were combined, concentrated, and dried in vacuo to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carbonitrile (2.57 g, 54% yield) as a yellow solid.

[0276] Step 3. (General procedure for nitrile hydrolysis using sulfuric acid) 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carbonitrile (7.11 g, 22.1 mmol) was dissolved in concentrated sulfuric acid (70 mL) and then stirred for 45 minutes. The reaction mixture was slowly poured onto crushed ice (500 cc), then placed in an ice bath and neutralized to pH 8-9 with concentrated NH4OH (approximately 190 mL) while maintaining the internal temperature below 35 °C. After stirring for 1 hour, the precipitate was filtered, washed with water, air-dried, and then further dried under vacuum by coevaporation with toluene and then dried under vacuum to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (7.5 g, quantitative yield) as a yellow solid.

[0277] Step 4. To a suspension of 4,6 - amino - 5-(3 - methoxy - 2,6 - dimethyl - phenyl)-2,3 - dimethyl - pyrrolo[2,3 - b]pyrazine - 7 - carboxamide (7.50 g, 22.1 mmol) in DCM (132 mL) was slowly added BBr3 (66.3 mmol, 6.4 mL). After stirring for 70 minutes, the reaction mixture was concentrated to dryness, redissolved in DCM, and MeOH was added (exotherm observed). After concentration, the crude mixture was co - evaporated with DCM / MeOH again, then carefully triturated with saturated aqueous NaHCO3 (100 mL), diluted with water, and stirred for 1.5 hours. The precipitate was filtered, washed with water, air - dried, and then purified by silica gel chromatography (dry packing) eluting with a gradient of 0 - 20% MeOH in DCM. The combined fractions were combined and re - purified by silica gel chromatography under the same conditions. The clean materials from both columns were combined, concentrated, and then dried under vacuum to give 6 - amino - 5-(3 - hydroxy - 2,6 - dimethyl - phenyl)-2,3 - dimethyl - pyrrolo[2,3 - b]pyrazine - 7 - carboxamide (5.3 g, 74% yield). 1 1H NMR (400 MHz, DMSO - d6) δ 9.57 (s, 1H), 7.45 (br s, 1H), 7.18 - 7.02 (m, 4H), 6.93 (d, J = 8.3 Hz, 1H), 2.48 - 2.45 (m, 3H), 2.35 - 2.24 (m, 3H), 1.81 - 1.73 (m, 3H), 1.68 (s, 3H). MS: [M + 1]: 326.4.

[0278] Compound 165 and Compound 166 were obtained by chiral SFC separation of Compound 164 (5.40 g, 16.6 mmol) (equipment: Waters Prep 100 SFC - MS; column: Phenomenex Lux Cellulose - 2, 30×250 mm, 5 μm; conditions: 45% CO2, isocratic with 1:1 55% ACN / EtOH; flow rate: 70 mL / min). [Chemical Structure] Compound 165 from chiral SFC separation of compound 164. Peak 1 (retention time 4.07 min, 99.99%): S-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (1.31 g) was obtained as a pale yellow-brown solid. 1 H NMR(400MHz,DMSO-d6)δ9.57(s,1H),7.45(br s,1H),7.12(br s,1H),7.09(br s,2H),7.06(d,J=8.8Hz,1H),6.93(d,J=8.3Hz,1H),2.47(s,3H),2.31(s,3H),1.76(s,3H),1.68(s,3H). MS:[M+1]:327.3. [ka] Compound 166 from chiral SFC separation of compound 164. Peak 2 (retention time 4.81 min, 99.83%): R-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (1.43 g) was obtained as a pale yellow-brown solid. 1 H NMR(400MHz,DMSO-d6)δ9.57(s,1H),7.45(br s,1H),7.12(br s,1H),7.09(br s,2H),7.07(d,J=8.4Hz,1H),6.93(d,J=8.3Hz,1H),2.47(s,3H),2.31(s,3H),1.76(s,3H),1.68(s,3H). MS:[M+1]:327.3.

[0279] [ka] Compound 173 (6-amino-2-cyclobutyl-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. A microwave vial was charged with [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (200 mg, 0.435 μmol) and Pd(PPh3)4 (56 mg, 49 μmol), flushed with nitrogen, added THF (2 mL), bubbled with nitrogen, added cyclobutylzinc bromide solution (0.5 M, 4.35 mL), bubbled with nitrogen, capped, and transferred to a heat block preheated to 70 °C. The reaction mixture was stirred for 90 min, cooled to room temperature, quenched with saturated aqueous NH4Cl, and extracted with EtOAc (2x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography eluting with a gradient of 0 to 100% EtOAc in hexanes to give 6-amino-2-cyclobutyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (110 mg, 69% yield) as a white / fawn solid.

[0280] Step 2. The same procedure used for compound 35 for OMe deprotection using BBr was carried out on the appropriate intermediate (110 mg, 0.301 mmol) to give a residue that was purified by preparative HPLC to give 6-amino-2-cyclobutyl-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (54 mg, 51% yield) as an off-white fluffy solid. 1 H NMR(400MHz,DMSO-d6)δ9.58(s,1H),7.61(s,1H),7.59(br s,1H),7.32(br s,2H),7.23(br s,1H),7.07(d,J=8.3Hz,1H),6.93(d,J=8.3Hz,1H),3.66(p,J=8.6Hz,1H),2.42-2. 17(m,4H),2.10-1.94(m,1H),1.88(td,J=8.5,4.0Hz,1H),1.76(s,3H),1.68(s,3H). MS:[M+1]:352.4.

[0281] [ka] Compound 178 (6-amino-2-(1-fluoro-1-methyl-ethyl)-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-(1-hydroxy-1-methyl-ethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (Compound 190, 46.0 mg, 0.129 mmol) in DCM (2 mL) was added Deoxo-fluor® solution (315 mg, 0.712 mmol, 50% in THF) dropwise at −78° C. The mixture was stirred at 0° C. for 45 minutes, concentrated, and purified by preparative HPLC to give 6-amino-2-(1-fluoro-1-methyl-ethyl)-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (23 mg, 50% yield) as an off-white fluffy solid. 1 H NMR(400MHz,DMSO-d6)δ9.63(s,1H),7.92(d,J=0.6Hz,1H),7.46(br s,2H),7.37(br s,1H),7.26(br s,1H),7.08(d,J=8.3Hz,1H),6.94(d,J=8.3Hz,1H),1.75(d,J=22.4Hz,6H),1.77(s,3H),1.69(s,3H). 19 F NMR (376MHz, DMSO-d6) δ-137.78 (hept, J=22.1Hz). MS:[M+1]:358.2.

[0282] Chiral SFC separation of compound 178 (19 mg, 0.053 mmol) (apparatus: Mettler Toledo Minigram SFC; column: Phenomenex Lux Cellulose-2, 10 × 250 mm, 5 μm; conditions: isocratic with 1:1 45% ACN / EtOH at 55% CO; flow rate: 10 mL / min) gave compounds 179 and 180. [ka] Compound 179 from chiral SFC separation of Compound 178. Peak 1 (retention time 3.63 min, 99.87%): S-6-Amino-2-(1-fluoro-1-methyl-ethyl)-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (5 mg) as a white cottony solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.63 (s, 1H), 7.92 (s, 1H), 7.46 (br s, 2H), 7.42 - 7.32 (m, 1H), 7.30 - 7.19 (m, 1H), 7.08 (d, J = 8.3 Hz, 1H), 6.94 (d, J = 8.2 Hz, 1H), 1.77 (s, 3H), 1.75 (d, J = 22.4 Hz, 6H), 1.69 (s, 3H). 19 F NMR (376 MHz, DMSO-d6) δ -137.75 (hept, J = 22.1 Hz). MS: [M + 1]: 358.2.

Chem.

[0283]

Chem.

[0284] Procedure 2. To an RBF containing DME (150 mL) with sodium hydride (3.13 g, 72.2 mmol, 60% w / w in mineral oil) was slowly added a solution of propanedinitrile (4.75 g, 71.9 mmol) in DME (50 mL). After stirring for 1 hour, 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyridine-2-amine (10.5 g, 28.7 mmol) and Pd(dppf)Cl2·DCM (2.31 g, 2.83 mmol) were added. The resulting mixture was degassed by bubbling N2 into the solution, fitted with a condenser, and heated to 95 °C for 1 hour. The reaction mixture was cooled to room temperature, poured into saturated aqueous NH4Cl, and extracted with DCM (3×). The combined organic extracts were washed with H2O, brine, dried over Na2SO4, filtered, and adsorbed onto silica. The crude residue was purified by silica gel chromatography (dry packing) eluting with a gradient of 0 - 100% EtOAc in heptane. Appropriate fractions were combined, concentrated, and the resulting solid was triturated with DCM, filtered, and dried under vacuum to give 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyrrolo[2,3-b]pyridine-3-carbonitrile (7.97 g, 79% yield) as a bright yellow solid. From the filtrate obtained from the previous trituration, a second crop was obtained by performing flash chromatography and trituration in the same manner, giving an additional 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyrrolo[2,3-b]pyridine-3-carbonitrile (1.04 g, 10% yield) as a dark yellow solid.

[0285] Step 3. To a solution of 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyrrolo[2,3-b]pyridine-3-carbonitrile (9.0 g, 25.6 mmol) in THF (120 mL) was added triethylamine (7.99 g, 78.9 mmol, 11 mL), DMAP (312 mg, 2.55 mmol), and tert-butoxycarbonyl tert-butyl carbonate (17.0 g, 77.9 mmol). The mixture was stirred at 50 °C for 40 min. Heating was stopped, ethylenediamine (6.20 g, 103 mmol, 6.90 mL) was added, and the mixture was stirred at room temperature for 45 min, then diluted with HO and DCM. The layers were separated, and the aqueous layer was extracted with DCM (2x). The combined organic extracts were washed with half-saturated brine, dried over Na2SO4, filtered, and concentrated. The crude residue was purified by silica gel chromatography eluting with a gradient of 0-60% EtOAc in heptane to give tert-butyl N-[3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyrrolo[2,3-b]pyridin-2-yl]carbamate (13.94 g, quantitative yield) as an off-white solid, which was obtained from tert-butyl N-[2-(tert-butoxycarbonylamino)ethyl]carbamate ( 1 H NMR showed contamination with 50 mol%.

[0286] Step 4. Palladium carbon (2.08 g, 1.95 mmol, 10% w / w), made as a slurry with some of the solvent mixture, was added to a RBF containing DCM (280 mL) and MeOH (280 mL) and N-[3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyrrolo[2,3-b]pyridin-2-yl]carbamic acid tert-butyl (13.94 g, 25.6 mmol), which was assumed to be quantitative and obtained from the previous step. H2 was passed through the reaction mixture and stirred overnight under a H2 atmosphere (balloon). N2 was passed through the reaction mixture, and it was filtered through a celite pad and rinsed with DCM and MeOH. The filtrate was concentrated and dried under vacuum to obtain a pale yellow solid, which was purified by silica gel chromatography (dry packing) eluting with a gradient of EtOAc (20 - 100%) in heptane. Appropriate fractions were combined and concentrated under vacuum to obtain N-[5-amino-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridin-2-yl]carbamic acid tert-butyl (9.34 g, 87% yield) as an off-white solid.

[0287] Step 5. To a solution of tert-butyl N-[5-amino-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridin-2-yl]carbamate (10.34 g, 24.5 mmol) in acetonitrile (100 mL) and DMF (60 mL) was added tert-butyl nitrite (5.20 g, 50.5 mmol, 6.0 mL), followed by copper(II) bromide (6.58 g, 29.4 mmol). The mixture was heated to 70 °C for 35 min, cooled to room temperature, diluted with HO (600 mL) and concentrated NHOH (30 mL), and extracted with EtOAc (3x). The combined organic extracts were washed with saturated NHCl (2x), half-saturated brine, dried over NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of 0-100% EtOAc in heptane. The appropriate fractions were combined and concentrated in vacuo to give tert-butyl N-[5-bromo-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridin-2-yl]carbamate (6.18 g, 52% yield) as an ivory solid.

[0288] Step 6. Tert-butyl N-[5-bromo-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridin-2-yl]carbamate (6.18 g, 12.7 mmol) in EtOH (60 mL) was treated with aqueous HCl (6 M, 34 mL) and stirred at 80 °C for 70 min, then cooled to room temperature and concentrated. The residue was dissolved in MeOH, made alkaline with excess EtN, and concentrated again. The residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of 0-100% EtOAc in heptane. The appropriate fractions were combined and concentrated in vacuo to give 2-amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridine-3-carbonitrile (3.70 g, 75% yield) as a dark red-purple solid.

[0289] Step 7. 2-Amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridine-3-carbonitrile (3.70 g, 9.6 mmol) was stirred in concentrated sulfuric acid (18 M, 25 mL) for 55 minutes, after which the reaction mixture was quenched with crushed ice, placed in an ice bath, and made alkaline to pH 8 - 9 by slowly adding saturated NH4OH. The resulting solid was collected by filtration on a Buchner funnel and washed with H2O. The material was air dried, then co-evaporated twice with toluene and dried under vacuum, then stirred in 10% MeOH / DCM and filtered through a silica plug eluting with 10% MeOH / DCM to remove residual ammonium salts. The filtrate was concentrated and then dried under vacuum to give 2-amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridine-3-carboxamide (3.80 g, 98% yield) as a peach-colored solid.

[0290] Procedure 8. A solution of methylmagnesium chloride (3 M, 18.8 mL) in THF (160 mL) within the RBF under N2 was added dropwise via an addition funnel with THF (0.5 M, 112 mL) containing zinc dichloride at room temperature. After addition, the resulting white suspension was stirred at room temperature for 35 minutes. 2-Amino-5-bromo-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridine-3-carboxamide (4.48 g, 11.1 mmol) was added to the zincate solution, the flask was rinsed with 20 mL of THF, and palladium(0) tetrakis(triphenylphosphine) (1.14 g, 0.987 mmol) was added. The mixture was bubbled with N2, then a condenser was attached and (with the heat block set to 80 °C) refluxed for 24 hours. The reaction mixture was cooled to room temperature, then diluted with saturated aqueous NH4Cl and extracted with EtOAc (3×). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (dry packing) eluting with a gradient of EtOAc in heptane (0 - 100%), then again purified by silica gel chromatography (dry packing) eluting with a gradient of MeOH in DCM (1 - 15%). The appropriate fractions from the two columns were combined and concentrated in vacuo to afford 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide (2.22 g, 59% yield, 77% purity) as a pale pink solid, which however contained some of the 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridine-3-carboxamide byproduct (19% by UPLCMS).

[0291] Procedure 9. A suspension containing 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide (2.22 g, 6.56 mmol, 77% purity) in DCM (25 mL) was added dropwise with DCM (1 M, 26 mmol, 26 mL) containing tribromoborane. The reaction mixture was stirred at room temperature for 45 minutes and then concentrated to dryness. The crude product was taken up in DCM, placed in an ice bath, and MeOH was carefully added (with exotherm). The mixture was concentrated to dryness and then co-evaporated with MeOH twice. The residue was triturated with saturated aqueous NaHCO3. The solid was collected by filtration on a Buchner funnel, washed with H2O, and air-dried. The still moist solid was dissolved in DCM / MeOH, concentrated to dryness, and triturated in 20% MeOH / DCM (50 mL). The solid was collected by filtration, washed with 20% MeOH / DCM, air-dried, and then dried under vacuum to give 2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide (1.60 g, 75% yield) as a pale yellowish brown solid. MS: [M+1]: 325.1. Different batches were purified by preparative HPLC to give 2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide (63% yield) as an off-white cottony solid. 1 1H NMR (400 MHz, DMSO-d6) δ 9.51 (s, 1H), 7.82 (s, 1H), 7.05 (d, J = 8.3 Hz, 1H), 6.90 (d, J = 8.2 Hz, 1H), 6.71 (br s, 2H), 6.64 (br s, 2H), 2.26 (s, 3H), 2.23 (s, 3H), 1.74 (s, 3H), 1.65 (s, 3H). MS: [M+1]: 325.1.

[0292] Chiral SFC separation of Compound 181 (1.60 g, 4.93 mmol) (Apparatus: Waters Prep 100 SFC-MS; Column: Phenomenex Lux Cellulose-2, 30×250 mm, 5 μm; Conditions: CO2 at 45% and 55% IPA + 10 mM ammonium formate with no gradient; Flow rate: 70 mL / min) gave Compound 182 and Compound 183. [Chemical formula] Compound 182 from the SFC separation of Compound 181. Peak 1 (Retention time 3.94 min, 99.86%): (S)-2-Amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide (381 mg) was obtained as an off-white cottony solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 7.83 (s, 1H), 7.05 (d, J = 8.3 Hz, 1H), 6.90 (d, J = 8.3 Hz, 1H), 6.72 (s, 2H), 6.65 (s, 2H), 2.26 (s, 3H), 2.24 (s, 3H), 1.74 (s, 3H), 1.65 (s, 3H). MS: [M+1]: 325.1. [Chemical formula] Compound 183 from the SFC separation of Compound 181. Peak 2 (Retention time 4.35 min, 98.09%): (R)-2-Amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide (495 mg) was obtained as an off-white cottony solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.50 (s, 1H), 7.83 (s, 1H), 7.05 (d, J = 8.2 Hz, 1H), 6.90 (d, J = 8.2 Hz, 1H), 6.72 (s, 2H), 6.66 (s, 2H), 2.26 (s, 3H), 2.24 (s, 3H), 1.74 (s, 3H), 1.65 (s, 3H). MS: [M+1]: 325.1.

[0293] [Chemical] Compound 181 (from Method O) 2-Amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide Step 1. Sulfuric acid (140.1 mL, 2575 mmol) was slowly added to water (1.15 L), and the solution was cooled to 25 °C. 2-Amino-3-bromo-5,6-dimethylpyridine (114.8 g, 571.0 mmol) was added all at once to obtain a solution. The solution was cooled to 0 - 5 °C in an ice-water bath to obtain a suspension. Under strong stirring, a solution of sodium nitrite (49.25 g, 713.7 mmol) in water (175.0 mL) was added dropwise over 90 minutes. The ice-water bath was removed, and the suspension was slowly warmed to 11 °C and stirred for 1 hour. A solution containing sodium hydroxide (175 g, 4.37 mol) in 400 mL of water was added dropwise while maintaining the temperature below 20 °C. The pH of the solution was adjusted to 7 with 70 mL of water containing K2HPO4 (about 58 g, 0.33 mol). The suspension was filtered at 10 °C. The filter cake was triturated in water (250 mL) and filtered. The filter cake was thoroughly washed with ice-cold water and dried by vacuum suction. The product was dried in an oven at 60 °C under vacuum overnight to obtain 3-bromo-5,6-dimethylpyridin-2-ol as a pale yellow crystalline solid (105.69 g, 91.6%). 1 H NMR (400 MHz, DMSO-d6) δ 11.96 (br s, 1H), 7.73 (s, 1H), 2.11 (s, 3H), 1.96 (s, 3H). MS: [M + 1]: 202.0, 204.0.

[0294] Step 2. To a solution of 3-bromo-5,6-dimethylpyridin-2-ol (105.3 g, 521.2 mmol) in N,N-dimethylformamide (316 mL) and toluene (527 mL) at 90° C. under nitrogen, phosphorus oxybromide (1.3:1, 56.5% (w / w) in xylene) (278 mL, 781.7 mmol) was added dropwise over 90 minutes. After the addition was complete, the mixture was stirred at 90° C. overnight. The mixture was cooled to room temperature and slowly added to water (2 L). The flask was washed with 500 mL of water. The combined aqueous phase was extracted with MTBE (3×1 L). The organic phases were combined and washed with 0.5 N NaOH (1 L), water (3 × 1 L), and brine (1 L), then dried over sodium sulfate and concentrated. The solid was partially dissolved in MTBE (400 mL) and heptane (300 mL) was added. The mixture was concentrated under reduced pressure to approximately 1.7 volumes to give a precipitate. The mixture was filtered and rinsed with heptane. The residue was dried to give 2,3-dibromo-5,6-dimethylpyridine as a pale yellow-brown solid (114.290 g, 82.8%). The filtrate was further concentrated and filtered to give a second crop of solid: (8.73 g, 6.32%). 1 H NMR (400MHz, DMSO-d6) δ7.95(s,1H), 2.36(s,3H), 2.21(s,3H). MS:[M+1]:264.0,266.0,268.0.

[0295] Step 3. A 2000 mL four-necked round-bottom flask was charged with Intermediate A2 (33.0 g, 218.0 mmol), degassed 1,2-dimethoxyethane (750 mL), 2,3-dibromo-5,6-dimethylpyridine (55 g, 207.6 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (10.8 g, 18.68 mmol), and cesium carbonate (169.1 g, 519.0 mmol). Nitrogen was injected into the suspension while the reaction mixture was sonicated for 20 minutes. Tris(dibenzylideneacetone)dipalladium(0) (8.55 g, 9.341 mmol) was added and the suspension was heated to reflux. After stirring for 13 hours, the reaction mixture was cooled to room temperature and filtered through a pad of silica gel. The filter cake was washed with ethyl acetate (1.2 L). The filtrate was evaporated to a volume of about 200 mL and heptane (300 mL) was added. The solvent was evaporated to give a suspension with a solvent volume of about 2 volumes. The suspension was filtered and washed with heptane to obtain 3-bromo-N-(3-methoxy-2,6-dimethylphenyl)-5,6-dimethylpyridin-2-amine as a pale yellow solid (56.2 g, 80.8%). 1 1H NMR (400 MHz, DMSO-d6) δ 7.57 (s, 1H), 7.26 (s, 1H), 7.02 (d, J = 8.6 Hz, 1H), 6.77 (d, J = 8.3 Hz, 1H), 3.76 (s, 3H), 2.07 (s, 3H), 2.04 (s, 3H), 2.03 (s, 3H), 1.94 (s, 3H). MS: [M+1]: 335.2, 337.2.

[0296] Procedure 4. Sodium tert-butoxide (46.3 g, 482.0 mmol) was added in four portions to a solution of degassed malononitrile (33.3 g, 503.5 mmol) in 1,2-dimethoxyethane (1000 mL). The reaction mixture was stirred at room temperature for 30 minutes to obtain a solution. 3-Bromo-N-(3-methoxy-2,6-dimethylphenyl)-5,6-dimethylpyridin-2-amine (80 g, 238.6 mmol) and a complex of 1,1'-bis(diphenylphosphino)ferrocene palladium(II) chloride with dichloromethane (14.9 g, 18.26 mmol) were added at once, and the suspension was heated to strong reflux. After stirring for 17 hours, the reaction mixture was transferred to a 5000 mL flask and ethyl acetate (1.5 L) was added. A solution containing N-acetyl-L-cysteine (12.1 g, 74 mmol, 4 times the molar content of Pd) and Na2CO3 (15.7 g, 148 mmol) in water (500 mL) was added. The biphasic solution was stirred at 60 °C for 10 minutes and then slowly cooled to 40 °C over 75 minutes. The layers were separated at 40 °C in a 5000 ml flask, and the organic phase was washed with water (2 × 250 mL) and brine (200 mL), and then filtered through a pad of silica gel (3 inches, 185 g). The filter cake was rinsed with DCM / EtOAc. The filtrate was evaporated, switching the solvent to EtOAc during rotavap evaporation to obtain a suspension. The suspension was filtered at room temperature, and the filter cake was triturated with 50 mL of ice-cold ethyl acetate. The product was filtered, and the filter cake was rinsed with 50 mL of ice-cold ethyl acetate. The product was dried overnight in an oven at 60 °C under vacuum to obtain 2-amino-1-(3-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile as a pale yellow solid. (65.38 g, 85.5%, 8% (w / w) of DME). 1 H NMR (400 MHz, DMSO-d6) δ 7.38 (s, 1H), 7.22 (d, J = 8.4 Hz, 1H), 7.07 (d, J = 8.6 Hz, 1H), 6.76 (br s, 2H), 3.84 (s, 3H), 2.26 (s, 3H), 2.23 (s, 3H), 1.78 (s, 3H), 1.69 (s, 3H). MS: [M+1]: 321.2.

[0297] Step 5. A solution of sulfuric acid (93 mL) / water (7.0 mL) was slowly added to methanesulfonic acid (600 mL, 9239 mmol) over 5 minutes at room temperature. 2-Amino-1-(3-methoxy-2,6-dimethylphenyl)-5,6-dimethyl-1H-pyrrolo[2,3-b]pyridine-3-carbonitrile (80 g, 249.7 mmol) was added in portions over 15 minutes while maintaining the reaction temperature below 40 °C. The resulting solution was stirred at room temperature for 90 minutes. DL-Methionine (149.028 g, 998.8 mmol) was added in portions over 20 minutes at below 40 °C. The solution was stirred at 40 °C. After stirring for 37 hours, the reaction mixture was cooled to room temperature, and then a solution containing K2HPO4 (100 g) and NaOH (540 g) in water (5 L) was slowly added over 1.5 hours. EtOAc (1 L) was added, and the biphasic mixture was stirred for 5 minutes to obtain a precipitate. The product was filtered. The mother liquor was extracted with EtOAc (3 × 1 L). The organic phases were combined, dried over Na2SO4, filtered, and concentrated under reduced pressure to approximately 100 mL to obtain a suspension. The suspension was filtered and washed with EtOAc (50 mL). The solids were combined and triturated twice in water (800 mL). The residue was suspended in EtOAc (500 mL), stirred for 10 minutes, and filtered. The product was dried in an oven under reduced pressure to obtain 67.8 g of a crude product. The compound was suspended in DMSO (350 mL, 5 volumes), and the mixture was heated to 65 °C to obtain a solution. The solution was slowly cooled to 28 °C in a water bath. Water (1.05 L) was added dropwise over 2 hours to obtain a suspension. After stirring at room temperature for 5 minutes, the product was filtered. The solid was triturated in 100 mL of water and filtered. The filter cake was washed with 2 × 100 mL of water. The product was dried in an oven at 60 °C under vacuum to obtain 2-amino-1-(3-hydroxy-2,6-dimethylphenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide as a pale yellow solid. 61.5 g, (75%, 3% (w / w) EtOAc and 6% (w / w) DMSO). 11H NMR (400 MHz, DMSO-d6) δ 9.47 (s, 1H), 7.82 (s, 1H), 7.05 (d, J = 8.2 Hz, 1H), 6.90 (d, J = 8.2 Hz, 1H), 6.71 (br.s, 2H), 6.64 (br.s., 2H), 2.27 (s, 3H), 2.24 (s, 3H), 1.75 (s, 3H), 1.66 (s, 3H). MS: [M+1]: 325.2.

[0298] [Chem.] Step 1. To a solution of Intermediate D (1.07 g, 2.33 mmol) in DCM (9 mL) was added a DCM solution of BBr3 (1 M, 9.3 mL, 9.3 mmol). The mixture was stirred at 0 °C for 2 h, silica was added, the mixture was concentrated, and then purified by silica gel chromatography (dry packing) eluting with a gradient of 0 - 20% MeOH in DCM to give [6-amino-7-carbamoyl-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (744 mg, 72% yield) as an off-white solid.

[0299] Step 2. A solution of [6-amino-7-carbamoyl-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl]trifluoromethanesulfonate (744 mg, 1.67 mmol) and PdCl(PPh) (117 mg, 0.167 mmol) in a mixture of DMF (8 mL), MeOH (8 mL), and EtN (1.40 mL, 10.0 mmol) was heated at 70 °C under a carbon monoxide atmosphere (balloon). The apparatus was pre-flushed with carbon monoxide once. After 2 h, additional PdCl(PPh) (117 mg, 0.167 mmol) was added, and the reaction mixture was allowed to continue for 18 h. The reaction mixture was cooled to room temperature, filtered through Celite, rinsed with MeOH, and the filtrate was concentrated. The residue was purified by silica gel chromatography (dry packing) eluting with a gradient of 0-20% MeOH in DCM to give a dark green sticky solid. The solid was dissolved in EtOAc and passed through a silica plug using 5% EtOAc / MeOH, and evaporation of the volatiles gave methyl 6-amino-7-carbamoyl-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2-carboxylate (418 mg, 70% yield) as a light brown sticky solid.

[0300] Procedure 3. A solution of methyl 3-amino-7-carbamoyl-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2-carboxylate (322 mg, 0.906 mmol) in THF (12 mL) was cooled to -40 °C, and a THF solution of MeMgCl (3 M, 4.53 mL, 13.1 mmol) was added dropwise. The mixture was allowed to warm to room temperature overnight, quenched with saturated aqueous NH4Cl (25 mL), adjusted to pH 7 - 8 using 1 N HCl, and the mixture was extracted with DCM (3×). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 20% MeOH in DCM to afford compound 190, 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-(1-hydroxy-1-methyl-ethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (103 mg, 32% yield) as a pale yellowish brown solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.61 (s, 1H), 7.99 (s, 1H), 7.50 (br s, 1H), 7.42 (br s, 2H), 7.23 (br s, 1H), 7.08 (d, J = 8.5 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 5.95 - 5.81 (m, 1H), 5.30 - 5.17 (m, 1H), 2.19 (s, 3H), 1.78 (s, 3H), 1.70 (s, 3H). MS: [M+1]: 338.1; Additionally, compound 184, 2-acetyl-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (31 mg, 10% yield) was obtained as a pale yellowish brown solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.83 (br s, 1H), 8.38 (d, J = 1.9 Hz, 1H), 7.68 (br s, 2H), 7.40 (s, 2H), 7.09 (d, J = 8.2 Hz, 1H), 6.97 (d, J = 8.1 Hz, 1H), 2.68 (s, 3H), 1.77 (s, 3H), 1.69 (s, 3H). MS: [M+1]: 340.1.

[0301] Chiral SFC separation of compound 190 (39 mg, 0.110 mmol) (apparatus: Mettler Toledo Minigram SFC; column: Phenomenex Lux Cellulose-2, 10 × 250 mm, 5 μm; conditions: isocratic with 40% IPA + 10 mM ammonium formate at 60% CO; flow rate: 10 mL / min) gave compounds 191 and 192. [ka] Compound 191 from chiral SFC separation of compound 190. Peak 1 (retention time 3.83 min, 100%): S-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-(1-hydroxy-1-methyl-ethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (10 mg) as an off-white solid. 1 H NMR(400MHz,DMSO-d6)δ9.61(br s,1H),7.99(s,1H),7.52(br d,J=3.0Hz,1H),7.32(br s,2H),7.18(br d,J=3.1Hz,1H),7.08(dt,J=8.2,0.8Hz,1H),6.93(d,J=8.3Hz,1H),5.26(s,1H),1.77(s,3H),1.68(s,3H),1.51(s,6H). MS:[M+1]:356.2. [ka] Compound 192 from chiral SFC separation of compound 190. Peak 2 (retention time 4.07 min): R-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-(1-hydroxy-1-methyl-ethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (10 mg). 11H NMR (400 MHz, DMSO-d6) δ 9.62 (br s, 1H), 7.99 (s, 1H), 7.52 (br d, J = 3.2 Hz, 1H), 7.32 (br s, 2H), 7.18 (br d, J = 3.1 Hz, 1H), 7.08 (dt, J = 8.3, 0.8 Hz, 1H), 6.93 (d, J = 8.3 Hz, 1H), 5.26 (s, 1H), 1.77 (s, 3H), 1.68 (s, 3H), 1.51 (s, 6H). MS: [M+1]: 356.2.

[0302] Compound 185 and compound 186 were obtained by chiral SFC separation of compound 184 (103 mg, 0.290 mmol) (apparatus: Mettler Toledo Minigram SFC; column: Phenomenex Lux Cellulose-2, 10 × 250 mm, 5 μm; conditions: 60% CO2, gradient-free with 40% IPA + 10 mM ammonium formate; flow rate: 10 mL / min).

Chemical formula

Chemical formula

[0303]

Chem.

[0304] Step 2. To a solution of 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[1-(trifluoromethyl)vinyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (42.0 mg, 0.104 mmol) in DCM (2 mL) was added a 0.5 M solution of diazomethane in EtO (400 μL) at 0 °C and then warmed to room temperature. A separately prepared solution of diazomethane (0.5 M, 400 μL) was added. After the reaction was deemed complete by UPLC MS, the reaction mixture was quenched with AcOH (200 μL), stirred for several minutes, concentrated to dryness, and then taken up in saturated aqueous NaHCO and DCM. The layers were separated (phase separator). The aqueous layer was extracted with DCM (3×). The combined organic extracts were concentrated and then dried under vacuum to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[5-(trifluoromethyl)-3,4-dihydropyrazol-5-yl]pyrrolo[2,3-b]pyrazine-7-carboxamide (48 mg, quantitative yield) as a yellow gum.

[0305] Step 3. 3.6-Amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[5-(trifluoromethyl)-3,4-dihydropyrazol-5-yl]pyrrolo[2,3-b]pyrazine-7-carboxamide (48.0 mg, 0.107 mmol) was dissolved in xylene (3 mL) and heated to 130 °C over a total of 75 minutes using a reflux condenser open to air. The reaction mixture was concentrated and then purified by silica gel chromatography eluting with a gradient of 0 - 100% EtOAc in heptane followed by a gradient of 0 - 20% MeOH in EtOAc to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (35 mg, 81% yield) as a pale yellow solid.

[0306] Step 4. For OMe deprotection using BBr3, the residue was obtained by the same procedure as used for compound 35 and purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (17 mg, 50% yield) as a white fluffy solid. 1 H NMR (400 MHz, DMSO-d6) δ 9.62 (s, 1H), 7.88 (s, 1H), 7.49 (br s, 2H), 7.36 (br s, 1H), 7.28 (br s, 1H), 7.08 (d, J = 8.2 Hz, 1H), 6.94 (d, J = 8.3 Hz, 1H), 1.76 (s, 3H), 1.69 (s, 3H), 1.46 - 1.29 (m, 4H). 19 F NMR (376 MHz, DMSO-d6) δ -66.85. MS: [M + 1]: 406.1.

[0307] Chiral SFC separation of compound 198 (14.5 mg, 0.358 mmol) (apparatus: Mettler Toledo Minigram SFC; column: Phenomenex Lux Cellulose-2, 10 × 250 mm, 5 μm; conditions: isocratic with 1:1 40% ACN / EtOH at 60% CO; flow rate: 10 mL / min) gave compounds 199 and 200. [ka] Compound 199 from chiral SFC separation of compound 198. Peak 1 (retention time 3.50 min, 99.99%): S-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (5 mg) as a white fluffy solid. 1H NMR(400MHz,DMSO-d6)δ9.61(s,1H),7.88(s,1H),7.48(br s,2H),7.36(br s,1H),7.27(br s,1H),7.08(d,J=8.3Hz,1H),6.94(d,J=8.3Hz,1H),1.76(s,3H),1.68(s,3H),1.43-1.33(m,4H). MS:[M+1]:406.2. [ka] Compound 200 from chiral SFC separation of compound 198. Peak 2 (retention time 3.81 min, 99.95%): R-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (5 mg) as a white fluffy solid. 1 H NMR(400MHz,DMSO-d6)δ9.61(s,1H),7.87(s,1H),7.48(br s,2H),7.36(br s,1H),7.27(br s,1H),7.08(d,J=8.3Hz,1H),6.94(d,J=8.3Hz,1H),1.76(s,3H),1.69(s,3H),1.43-1.35(m,4H). MS:[M+1]:406.2.

[0308] [ka] Compound 209 (5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carbonitrile (298 mg, 0.927 mmol) in THF (7 mL) was added tert-butyl nitrite (550 μL, 4.63 mmol). After stirring for 30 min, the mixture was refluxed for 3.5 h, then cooled to room temperature, concentrated to dryness, and purified by silica gel chromatography eluting with a gradient of 0 to 100% EtOAc in hexanes. The combined pure fractions were concentrated and dried under vacuum to give 5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carbonitrile (150 mg, 53% yield) as a pale yellow solid.

[0309] Step 2. The same procedure used for compound 164 for nitrile hydrolysis using sulfuric acid was carried out on the appropriate intermediate (150 mg, 0.490 mmol) to give 5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (144 mg, 91% yield) as an off-white solid.

[0310] Step 3. The same procedure used for OMe deprotection using BBr3 as used for compound 35 gave a residue which was purified by preparative HPLC to give 5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (76 mg, 55% yield) as an off-white fluffy solid. 1H NMR (400 MHz, DMSO-d6) δ 9.65 (br s, 1H), 8.13 (s, 1H), 7.86 (br s, 1H), 7.60 (br s, 1H), 7.05 (d, J = 8.2 Hz, 1H), 6.92 (d, J = 8.3 Hz, 1H), 2.64 (s, 3H), 1.75 (s, 3H), 1.64 (s, 3H). One Me singlet is probably buried by the DMSO peak. MS: [M+1]: 311.1.

[0311] [ka] Compound 212 (2-amino-1-(5-hydroxy-2-methyl-phenyl)-6-(trifluoromethyl)pyrrolo[3,2-b]pyridine-3-carboxamide) Step 1. To a suspension of NaH (108 mg, 2.83 mmol, 60% dispersion in mineral oil) in DME (3 mL) was added 3-bromo-2-chloro-5-(trifluoromethyl)pyridine (300 mg, 1.15 mmol) in DME (3 mL) dropwise. After the addition, the mixture was stirred for 25 minutes, after which propanedinitrile (188 mg, 2.85 mmol) was added. The resulting mixture was refluxed for 18 hours, cooled to room temperature, and concentrated in vacuo. The residue was purified by preparative HPLC to give 2-[3-bromo-5-(trifluoromethyl)-2-pyridyl]propanedinitrile (100 mg, 30% yield).

[0312] Step 2. To a solution of [[3-bromo-5-(trifluoromethyl)-2-pyridyl]propanedinitrile (50 mg, 172 μmol) in DMF (2 mL) were added Pd2dba3 (16 mg, 17 μmol), 5-(methoxymethoxy)-2-methylaniline (33.3 mg, 199 μmol), Cs2CO3 (84 mg, 259 μmol) and Xantphos (10.0 mg, 17.3 μmol). The mixture was degassed under vacuum and backfilled with nitrogen (3 times). The mixture was stirred at 130 °C for 8 hours, cooled to room temperature, diluted with water and extracted with EtOAc (3×). The combined organic extracts were washed with brine, dried over Na2SO4, filtered and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 5 - 100% EtOAc in hexane to give 2-amino-1-[5-(methoxymethoxy)-2-methyl-phenyl]-6-(trifluoromethyl)pyrrolo[3,2-b]pyridine-3-carbonitrile (30 mg, 46% yield).

[0313] Step 3. 2-Amino-1-[5-(methoxymethoxy)-2-methyl-phenyl]-6-(trifluoromethyl)pyrrolo[3,2-b]pyridine-3-carbonitrile (30 mg, 135 μmol) was stirred in H2SO4 (1 mL). After 90 minutes, the reaction mixture was poured onto crushed ice, placed in an ice bath and neutralized with 1:1 NH4OH / H2O. The precipitate was filtered, washed with water and air dried overnight. Purification by preparative HPLC gave 2-amino-1-(5-hydroxy-2-methyl-phenyl)-6-(trifluoromethyl)pyrrolo[3,2-b]pyridine-3-carboxamide (3.2 mg, 11% yield) as an orange solid. 1 1H NMR (400 MHz, DMSO-d6) δ 9.76 (s, 1H), 8.45 (s, 1H), 7.77 (s, 1H), 7.34 (s, 2H), 7.31 - 7.13 (m, 2H), 6.98 (s, 1H), 6.91 (d, J = 8.5 Hz, 1H), 6.71 (s, 1H), 1.78 (s, 3H). MS: [M+1]: 351.3.

[0314]

Chemical Structure

[0315] Step 2. To a solution of 6-amino-3-bromo-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (300 mg, 739 μmol) and CsCO (440 mg, 1.35 mmol) in DMF (3 mL) was added 1,1,1-trifluoro-N-phenyl-N-(trifluoromethylsulfonyl)methanesulfonamide (288 mg, 807 μmol). The mixture was stirred for 1 h, diluted with water, and extracted twice with EtOAc. The combined organic extracts were washed with brine, dried over NaSO, filtered, and concentrated. The residue was purified by silica gel chromatography using a gradient of 0 to 100% EtOAc in hexanes to give 6-amino-3-bromo-7-carbamoyl-5-(3-methoxy-2,6-dimethylphenyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl trifluoromethanesulfonate (460 mg, 43% yield).

[0316] Step 3. A microwave vial charged with [6-amino-3-bromo-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (35 mg, 65 μmol), ethynylcyclopropane (5.0 mg, 75 μmol), CuI (1.3 mg, 7 μmol), and PdCl(PPh) (5.0 mg, 7 μmol) in DMF (1 mL) was flushed with nitrogen, then EtN (520 μmol, 73 μL) was added and the mixture was stirred for 1 h. The mixture was filtered and purified by preparative HPLC to give trifluoromethanesulfonic acid [6-amino-7-carbamoyl-3-(2-cyclopropylethynyl)-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] (10 mg, 29% yield).

[0317] Step 4. A mixture of tributyl(thiazol-2-yl)stannane (38.5 μmol, 12.1 μL), trifluoromethanesulfonate [6-amino-7-carbamoyl-3-(2-cyclopropylethynyl)-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] (10.0 mg, 19.1 μmol), CuI (0.5 mg, 2.5 μmol), LiCl (1.7 mg, 40 μmol), and PdCl(dppf).CHCl (1.5 mg, 2 μmol) in DMF (3 mL) was degassed under vacuum and then backfilled with nitrogen. The reaction mixture was stirred at 120 °C for 3 h. Water was added and the resulting precipitate was removed by filtration to give crude 6-amino-3-(2-cyclopropylethynyl)-5-(3-methoxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (5 mg, 57% yield).

[0318] Procedure 5. For the deprotection of OMe using BBr3, a residue was obtained by the same procedure as that used for Compound 35, and this was purified by preparative HPLC to give 6-amino-3-(2-cyclopropylethynyl)-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (1.1 mg, 23% yield). 1 1H NMR (400 MHz, methanol-d4) δ 9.16 (d, J = 4.2 Hz, 1H), 8.47 (d, J = 4.2 Hz, 2H), 7.89 (d, J = 1.1 Hz, 1H), 7.15 (d, J = 8.3 Hz, 1H), 6.98 (d, J = 8.3 Hz, 1H), 2.55 - 2.47 (m, 1H), 1.88 (d, J = 21.7 Hz, 6H), 1.37 - 1.31 (m, 2H), 1.11 - 1.05 (m, 2H). MS: [M+1]: 445.3.

[0319]

Chemical Structure

[0320] Step 2. For OMe deprotection using BBr3, a residue was obtained by the same procedure as used for Compound 35 and purified by preparative HPLC to obtain 2-amino-1-(3-hydroxy-2,6-dimethylphenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carboxamide (7 mg, 45% yield). 1 H NMR (400 MHz, DMSO-d6) δ 9.63 (s, 1H), 7.70 (s, 2H), 7.35 - 7.13 (m, 2H), 7.05 (d, J = 8.3 Hz, 1H), 6.92 (d, J = 8.3 Hz, 1H), 3.62 - 3.51 (m, 11H), 3.49 (s, 2H), 2.52 (q, J = 4.3, 3.9 Hz, 4H), 1.74 (s, 3H), 1.66 (s, 3H). MS: [M + 1]: 544.5.

[0321]

Chemical Structure

[0322] Procedure 2. For OMe deprotection using BBr₃, a residue was obtained by the same procedure as used for compound 35 and purified by preparative HPLC to give compound 219, 6-amino-2-cyano-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (2.6 mg, 9% yield), which 1 had ¹H NMR (400 MHz, DMSO-d₆) δ 9.64 (s, 1H), 8.28 (s, 1H), 7.88 (s, 2H), 7.38 (s, 1H), 7.05 (d, J = 8.4 Hz, 2H), 6.92 (d, J = 8.3 Hz, 1H), 1.73 (s, 3H), 1.65 (s, 3H), MS: [M + 1]: 324.2; and further, compound 220, 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2,7-dicarboxamide (10 mg, 33% yield) was obtained. 1 ¹H NMR (400 MHz, DMSO-d₆) δ 9.57 (s, 1H), 8.67 (s, 1H), 8.35 (s, 1H), 7.64 (s, 3H), 7.41 (s, 1H), 7.13 (s, 1H), 7.11 - 6.97 (m, 1H), 6.91 (d, J = 8.3 Hz, 1H), 1.73 (s, 3H), 1.65 (s, 3H). MS: [M + 1]: 341.4.

[0323]

Chemical formula

[0324] Step 2. To a solution of propanedinitrile (556 mg, 8.41 mmol) in DME (20 mL) was added NaH (361 mg, 8.34 mmol, 60% dispersion in mineral oil). The mixture was stirred for 5 minutes, after which 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyridin-2-amine (1.55 g, 4.13 mmol) and Pd(PPh3)4 (231 mg, 200 μmol) were added. The resulting mixture was stirred in a pressure vial at 120 °C for 17 hours. The DME was removed under reduced pressure, after which the mixture was diluted with EtOAc, washed with water, brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash chromatography eluting with a gradient of 0 to 100% EtOAc in hexanes to give 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carbonitrile (44 mg, 3% yield).

[0325] Step 3. For the nitrile hydrolysis using sulfuric acid, the same procedure as used for Compound 164 was carried out on the appropriate intermediate (44 mg, 0.121 mmol) to give 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carboxamide (40 mg, 87% yield).

[0326] Step 4. For the OMe deprotection using BBr3, the residue was obtained by the same procedure as used for Compound 35 and purified by preparative HPLC to give 2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carboxamide (25 mg, 59% yield). 1 H NMR (400 MHz, DMSO-d6) δ 9.66 (s, 1H), 8.47 (dt, J = 2.0, 1.0 Hz, 1H), 7.77 (s, 1H), 7.31 (s, 2H), 7.23 (s, 1H), 7.10 (dt, J = 8.3, 0.8 Hz, 1H), 7.00 - 6.80 (m, 2H), 1.82 - 1.57 (m, 6H). MS: [M+1]: 365.3.

[0327]

Chemical Structure

[0328]

Chemical Structure

[0329] Step 2. To methyl 6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2-carboxylate (689 mg, 1.87 mmol) in THF (5 mL) was added 1 M NaOH (5.60 mL), and the mixture was stirred for 1.5 h. The pH was acidified using concentrated HCl, DMSO was added, the volatiles were removed under reduced pressure, and the residue was purified by preparative HPLC to give 6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2-carboxylic acid (366 mg, 55% yield).

[0330] Step 3. To a solution of pyridin-3-amine (7.95 mg, 84.4 μmol), 6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2-carboxylic acid (25 mg, 70 μmol), and HATU (29 mg, 77 μmol) in DCM (5 mL) was added DIPEA (211 μmol, 37 μL). The reaction mixture was stirred for 18 h. Water was added to the mixture, and the organic phase was separated, dried over NaSO, filtered, and concentrated in vacuo to give crude 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-N-(3-pyridyl)pyrrolo[2,3-b]pyrazine-2,7-dicarboxamide (21 mg, 34% yield, 49% purity).

[0331] Step 4. The same procedure used for compound 35 for OMe deprotection using BBr3 gave a residue which was purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-N2-(3-pyridyl)pyrrolo[2,3-b]pyrazine-2,7-dicarboxamide (3.5 mg, 12% yield). 1H NMR(400MHz,DMSO-d6)δ10.82(s,1H),9.61(s,1H),8.92(d,J=2.5Hz,1H),8.49(s,1H),8.32(dd,J=4.8,1.6Hz,1H),8.20-8.10(m,1H), 7.75(d,J=9.0Hz,3H),7.40(dd,J=8.3,4.7Hz,1H),7.30(s,1H),7.07(d,J=8.3Hz,1H),6.93(d,J=8.3Hz,1H),1.75(s,3H),1.67(s,3H). MS:[M+1]:418.3.

[0332] [ka] Compound 257 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-vinyl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. A mixture of tributyl(vinyl)stannane (37.1 mg, 117 μmol), Intermediate H (40.0 mg, 106 μmol), CuI (2.56 mg, 13.4 μmol), LiCl (9.30 mg, 220 μmol), and PdCl(dppf).CHCl (8.1 mg, 10 μmol) in DMF (1 mL) was degassed under vacuum and then backfilled with nitrogen. The final mixture was stirred at 130 °C for 3 h, cooled to room temperature, filtered, and purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-vinyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (1.4 mg, 4% yield). 1 H NMR(400MHz,DMSO-d6)δ9.59(s,1H),8.29(s,1H),8.19(s,1H),7.38(d,J=24.4Hz,2H),7.20(s,1H),7.05(d,J=8.3Hz,1H),6.91(d, J=8.3Hz,1H),6.69(dd,J=17.3,10.8Hz,1H),5.80(dd,J=17.3,1.8Hz,1H),5.15(dd,J=10.7,1.8Hz,1H),1.75(s,3H),1.67(s,3H). MS:[M+1]:324.3.

[0333] [ka] Compound 260 (6-amino-2-(cyclopropoxy)-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. A solution of cyclopropanol (12 mg, 202 μmol), intermediate D (31 mg, 67 μmol), and CsCO (66 mg, 202 μmol) in NMP (1 mL) was stirred at 140° C. for 16 h. The mixture was cooled to room temperature, filtered, and purified by preparative HPLC to give 6-amino-2-(cyclopropoxy)-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (3 mg, 12% yield).

[0334] Step 2. The same procedure used for OMe deprotection using BBr3 as used for compound 35 gave a residue which was purified by preparative HPLC to give 6-amino-2-(cyclopropoxy)-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (0.52 mg, 18% yield). 1 H NMR(400MHz,DMSO-d6)δ9.57(s,1H),8.45(s,1H),7.35(s,1H),7.19(d,J=19.0Hz,3H),7.02(d,J=8.3Hz,1 H),6.88(d,J=8.3Hz,1H),4.25-4.14(m,1H),1.73(s,3H),1.65(s,3H),0.73(t,J=5.0Hz,2H),0.68(s,2H). MS:[M+1]:354.4.

[0335] [ka] Compound 263 (6-amino-2-(difluoromethyl)-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. A microwave flask was charged with PdCl(PPh) (79.6 mg, 109 μmol), Intermediate D (1.00 g, 2.18 mmol), and sodium formate (222 mg, 3.27 mmol). The flask was flushed with carbon monoxide. DMF (5 mL) was added, and a slow stream of carbon monoxide was passed through the suspension. The mixture was vigorously stirred under an atmosphere of carbon monoxide at 100 °C for 2 h. The resulting mixture was cooled to room temperature, filtered, and the supernatant was purified by preparative HPLC to give a crude mixture of 6-amino-2-formyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (556 mg, 75% yield).

[0336] Step 2. To a solution of 6-amino-2-formyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (180 mg, 530 μmol) in DCM (2 mL) was added dropwise Deoxo-Fluor® solution (50% in THF, 1.46 M, 2.00 mL) at 0° C. The mixture was allowed to warm to room temperature. After 2 hours, excess Deoxo-Fluor® solution (50% in THF, 1.17 g, 2.65 mmol) was added. After 1 hour, additional Deoxo-Fluor® solution (50% in THF, 2.35 g, 5.30 mmol) was added, and the final mixture was stirred for 4 hours, after which it was diluted with DCM and saturated aqueous Na2CO3 solution was added. The biphasic mixture was stirred for 1 hour. The organic layer was separated, dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC to give 6-amino-2-(difluoromethyl)-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (10 mg, 5% yield).

[0337] Step 3. The same procedure used for compound 35 for OMe deprotection using BBr3 gave a residue which was purified by preparative HPLC to give 6-amino-2-(difluoromethyl)-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (3.0 mg, 31% yield). 11H NMR (400 MHz, DMSO-d6) δ 9.66 (s, 1H), 8.29 (s, 1H), 8.00 (s, 1H), 7.65 (s, 2H), 7.30 (d, J = 34.1 Hz, 2H), 7.11 - 6.98 (m, 2H), 6.99 - 6.69 (m, 1H), 1.73 (s, 3H), 1.65 (s, 3H). MS: [M+1]: 348.2.

[0338] [Chemical formula] Compound 266 (6-Amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-bis(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. Iodine (33 mg, 130 μmol) was added to a suspension containing flaky magnesium (380 mg, 15.7 mmol) in Et2O (20 mL). The mixture was stirred for 10 min, and then CD3I (975 μL, 15.7 mmol) was added. The mixture was stirred under a nitrogen atmosphere for 18 h to produce an off-white suspension. ZnCl2 (0.5 M in THF, 1.4 mL) was added dropwise, and the mixture was stirred for 20 min. Intermediate D (1.2 g, 2.61 mmol) and Pd(PPh3)4 (300 mg, 259 μmol) were added. The mixture was stirred at 70 °C for 72 h under a nitrogen atmosphere. The reaction was quenched with 1 M aqueous HCl, diluted with water, and extracted twice with EtOAc. The combined organic extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 80% EtOAc in hexane to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (400 mg, 46% yield).

[0339] Step 2. To a solution of 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (400 mg, 1.22 mmol) in DMF (2 mL) was added NBS (259 mg, 1.46 mmol). The mixture was stirred for 10 minutes, diluted with water, stirred for 20 minutes, and filtered. The precipitate was purified by preparative HPLC to give 6-amino-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)-2-(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (400 mg, 81% yield).

[0340] Step 3. To a suspension of magnesium (177 mg, 7.3 mmol) in ether (10 mL) was added iodine (16 mg, 61 μmol). The mixture was stirred for 10 minutes, and then CD3I (455 μL, 7.31 mmol) was added thereto. The mixture was stirred for 18 hours under a nitrogen atmosphere to obtain an off-white suspension. ZnCl2 (0.5 M in THF, 14.6 mL) was added dropwise. After the addition, the mixture was stirred for 20 minutes. 6-Amino-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)-2-(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (496 mg, 1.22 mmol), Pd2dba3 (111 mg, 122 μmol), and tri-tert-butylphosphonium tetrafluoroborate (71 mg, 244 μmol) were added, and the mixture was stirred at 70 °C for 18 hours under a nitrogen atmosphere. The reaction was quenched with 1 M aqueous HCl, diluted with water, and extracted twice with EtOAc. The combined organic extracts were washed with water, brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 60% EtOAc in hexane to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-bis(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (400 mg, 95% yield).

[0341] Procedure 4. For the deprotection of OMe using BBr3, the residue was obtained by the same procedure as used for compound 35, and purified by preparative HPLC to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-bis(trideuteromethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (65 mg, 17% yield). 1 1H NMR (400 MHz, DMSO-d6) δ 9.55 (s, 1H), 7.41 (s, 1H), 7.22 - 6.98 (m, 4H), 6.89 (d, J = 8.3 Hz, 1H), 1.74 - 1.69 (m, 3H), 1.63 (s, 3H). MS: [M + 1]: 332.2.

[0342] Preparation Example of Arylamine The compounds of the present invention were prepared using various arylamines. Some of these arylamines were commercially available and some were prepared. Some examples of such arylamines described herein for preparation are listed in Table 2. [Table 2]

[0343] Preparation of Arylamine A1 The compounds of the present invention can be prepared from arylamine A1 shown in Scheme A1 and prepared as described herein. Commercially available 4-methyl-3-nitro-phenol can have its O protected by a suitable protecting group, such as O-MOM. The nitro can be reduced to produce arylamine A1. [Chemical formula]

[0344] Step 1. To a suspension of 4-methyl-3-nitro-phenol (25 g, 163 mmol) in DCM (250 mL) was added DIPEA (34 mL, 195 mmol), followed by dropwise addition of chloro(methoxy)methane (26.0 g, 323 mmol, 24.5 mL). After stirring for 18 h, the reaction mixture was washed with water. The layers were separated. The organic layer was washed with 0.2 N HCl (2×), brine, dried over MgSO4, filtered, concentrated, and then dried under vacuum to afford 4-(methoxymethoxy)-1-methyl-2-nitro-benzene (31.4 g, 98% yield) as a dark red oil.

[0345] Step 2. To a suspension of 4-(methoxymethoxy)-1-methyl-2-nitro-benzene (31.4 g, 159 mmol) in EtOH (200 mL) and water (75 mL) was added ammonium chloride (43.3 g, 809 mmol), followed by addition of iron powder (44.5 g, 796 mmol). The reaction mixture was heated to 80 °C over 3.5 h and then warmed to 90 °C and stirred for 4 days. The reaction mixture was cooled to room temperature, filtered, and rinsed with EtOAc. The filtrate was concentrated, diluted with EtOAc and saturated aqueous NaHCO3. The layers were separated and the aqueous layer was back-extracted with EtOAc (2×). The combined organic extracts were washed with brine, dried over MgSO4, filtered, concentrated to give 26.3 g of a crude product as a dark brown oil, which was purified by elution through a silica gel pad with 20 - 30% EtOAc in hexane. The pure fractions were combined, concentrated, and then dried under vacuum to afford 5-(methoxymethoxy)-2-methyl-aniline (25.4 g, 95% yield) as a purple oil. 1 1H NMR (400 MHz, chloroform-d) δ 6.94 (dd, J = 8.0, 1.7 Hz, 1H), 6.45 - 6.30 (m, 2H), 5.12 (s, 2H), 3.47 (s, 3H), 2.10 (s, 3H). MS: [M + 1]: 168.3.

[0346] Preparation of Aryl Amine A2 The compounds of the present invention can be prepared from arylamine A2, which can be prepared as described herein, as shown in Scheme A2 (adapted from Can J Chem 2012, 90, 75-84). Commercially available 1,3-dimethyl-2-nitrobenzene can be brominated under suitable bromination conditions. The resulting bromo can be converted to methoxy by treatment with sodium methoxide and copper(I) bromide. The nitro can be reduced to generate arylamine A2. [ka]

[0347] Step 1. A 3 L, three-necked round-bottom flask equipped with a mechanical stirrer, reflux condenser, and addition funnel was charged with 1,3-dimethyl-2-nitrobenzene (300 g, 1.98 mol), DCM (900 mL), iron powder (28.0 g, 501 mmol), and iron(III) bromide (11.9 g, 40.3 mmol). Bromine (112 mL, 2.19 mol) was added dropwise via the addition funnel over 45–60 min. Internal temperature monitoring indicated an exotherm to 30 °C. Ninety minutes after the complete bromine addition, additional bromine (5 mL, 97.6 mmol) was added, and the reaction mixture was stirred for another 45 min to complete the conversion. The reaction mixture was diluted with ice water (1.5 L) and EtO (1.5 L). The layers were separated. The aqueous layer was back-extracted with EtO (0.5 L). The combined organic layers were washed with 20% aqueous NaSO (1 L), brine (500 mL), dried over NaSO, filtered through a silica gel pad (300 cc), concentrated, and then dried under vacuum to give 1-bromo-2,4-dimethyl-3-nitro-benzene (451.5 g, 99% yield) as an off-white solid.

[0348] Step 2. A 5 L, four-necked round-bottom flask equipped with a mechanical stirrer and reflux condenser was charged with 1-bromo-2,4-dimethyl-3-nitrobenzene (451.5 g, 1.96 mol) in DMF (1.6 L). CuBr (28.0 g, 195 mmol) was added, followed by MeONa (1.31 L, 5.89 mol, 25% in MeOH). The reaction mixture was slowly heated to 95 °C to achieve a gentle reflux. After 6 h, the reaction mixture was allowed to cool to room temperature overnight. The reaction mixture was diluted with EtO and saturated aqueous NH4Cl (1.5 L each). The layers were separated, and the aqueous layer was back-extracted with EtO (750 mL). The combined organic extracts were washed with brine (750 mL), dried over Na2SO4, filtered through a silica pad, rinsed with Et2O, concentrated, and dried under vacuum to give 1-methoxy-2,4-dimethyl-3-nitro-benzene (352 g, 99% yield) as an ochre solid.

[0349] Step 3. To a solution of 1-methoxy-2,4-dimethyl-3-nitro-benzene (115 g, 635 mmol) in EtOH (1.5 L) in a 3 L three-neck flask equipped with a mechanical stirrer, iron powder (213 g, 3.81 mol) was added, followed by the addition of a solution of ammonium chloride (204 g, 3.81 mol) in water (500 mL) in several portions. The mixture was heated to 85 °C for 8 h. The mixture was cooled to room temperature and filtered through Celite. The volume of the filtrate was reduced (most of the EtOH was evaporated), and the resulting mixture was diluted with EtO (800 mL) and water (150 mL). The layers were separated, and the aqueous layer was back-extracted with EtO (500 mL). The combined organic extracts were washed with brine, dried over Na.sub.2SO.sub.4, filtered, concentrated, and dried under vacuum to give 3-methoxy-2,6-dimethyl-aniline (89.1 g, 93% yield) as a brown oil. 1 H NMR (400 MHz, chloroform-d) δ 6.88 (dq, J = 8.3, 0.7 Hz, 1H), 6.31 (d, J = 8.2 Hz, 1H), 3.79 (s, 3H), 3.61 (br s, 2H), 2.14 (d, J = 0.7 Hz, 3H), 2.07 (s, 3H). MS: [M+1]: 152.3.

[0350] Preparation of Aryl Amine A3 The compounds of the present invention can be prepared from aryl amine A3, which is shown in Scheme A3 and can be prepared as described herein. The methoxy group of 1-methoxy-2,4-dimethyl-3-nitro-benzene described in the preparation of intermediate A2 can be cleaved using BBr3, and the resulting phenol can have its O protected by a suitable protecting group, such as O-MOM. The nitro can be reduced to produce aryl amine A3.

Chemical formula

[0351] Step 1. To a solution of 1-methoxy-2,4-dimethyl-3-nitro-benzene (20 g, 110 mmol) in DCM (200 mL) cooled in a dry ice / acetonitrile bath, a solution of BBr3 in DCM (1 M, 168 mL) was added dropwise via an addition funnel. The mixture was slowly warmed to room temperature overnight. The reaction mixture was then slowly poured into a stirred mixture of ice and water (1 L), KH2PO4 (75 g). The layers were separated, and the aqueous layer was extracted with DCM (2 × 500 mL). The combined organic extracts were washed with brine (500 mL), dried over MgSO4, filtered through a silica pad (375 g) eluting with DCM, concentrated, and dried under vacuum to obtain 2,4-dimethyl-3-nitro-phenol (18.0 g, 98% yield) as a yellow solid.

[0352] Step 2. To a suspension containing 2,4-dimethyl-3-nitro-phenol (18.96 g, 113 mmol) in DCM (200 mL) was added dropwise DIPEA (23.7 mL, 136 mmol), and then chloro(methoxy)methane (9.5 mL, 125 mmol) was added dropwise. After stirring for 3.5 h, additional chloro(methoxy)methane (2.0 mL, 26 mmol) was added and the reaction mixture was stirred overnight. The reaction mixture was quenched with saturated aqueous NH4Cl (100 mL) and diluted with water (100 mL). The layers were separated and the aqueous layer was back-extracted with DCM (100 mL). The combined organic extracts were washed with 0.2 N HCl (2×100 mL), 1 M NaOH (100 mL), brine (100 mL), dried over MgSO4, filtered through silica (ca. 100 cc), eluted with DCM, concentrated and dried under vacuum to give 1-(methoxymethoxy)-2,4-dimethyl-3-nitro-benzene (22.1 g, 92% yield) as a pale yellow waxy solid.

[0353] Step 3. To a flask under nitrogen containing palladium carbon (5.06 g, 4.76 mmol, 10% w / w) was added MeOH (300 mL), followed by 1-(methoxymethoxy)-2,4-dimethyl-3-nitro-benzene (20.1 g, 95.1 mmol). Hydrogen was bubbled through the flask and the mixture was stirred under a hydrogen atmosphere for 2 days. Nitrogen was bubbled through the reaction mixture for 2 h and celite was added. The mixture was filtered through a celite pad using MeOH and DCM. The filtrate was concentrated and dried under vacuum to give 3-(methoxymethoxy)-2,6-dimethyl-aniline (17.1 g, 99% yield) as a pale orange cloudy oil. 1 1H NMR (400 MHz, chloroform-d) δ 6.86 (d, J = 8.3 Hz, 1H), 6.49 (d, J = 8.3 Hz, 1H), 5.15 (s, 2H), 3.48 (s, 3H), 2.14 (s, 3H), 2.11 (s, 3H). MS: [M+1]: 182.2.

[0354] Preparation of Aryl Amine A4 The compounds of the present invention can be prepared from arylamine A4, which can be prepared as shown in Scheme A4 and described herein. Commercially available 2-chloro-3-methoxybenzoic acid can be brominated with a suitable brominating reagent, and the carboxylic acid can be converted to NHBoc under Curtius conditions. The bromo can be converted to methyl, and the NHBoc can be cleaved under acidic conditions to generate arylamine A4. [ka]

[0355] Step 1. To a solution of 2-chloro-3-methoxy-benzoic acid (50 g, 268 mmol) in AcOH (250 mL) and water (250 mL) was added bromine (27.5 mL, 537 mmol) dropwise. The mixture was stirred at 60 °C for 18 h, cooled to room temperature, brine was added, and the mixture was extracted twice with DCM. The combined organic extracts were dried over NaSO, filtered, and concentrated in vacuo to give 6-bromo-2-chloro-3-methoxy-benzoic acid (71 g, quantitative yield) as a brown oil that solidified upon standing under vacuum over the weekend.

[0356] Step 2. To a solution of 6-bromo-2-chloro-3-methoxy-benzoic acid (23.6 g, 88.9 mmol), EtN (38 mL, 271 mmol), and tert-butanol (42.5 mL, 450 mmol) in toluene (500 mL) was added [azido(phenoxy)phosphoryl]oxybenzene (29.5 mL, 136 mmol). The mixture was heated at 100 °C for 16 h, cooled to room temperature, and then the volatiles were removed in vacuo. The residue was diluted with EtOAc (100 mL), and the organic layer was washed with 5% citric acid, water, saturated aqueous NaHCO, brine, dried over NaSO, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with a gradient of 0-20% EtOAc in hexanes to give tert-butyl N-(6-bromo-2-chloro-3-methoxy-phenyl)carbamate (16.2 g, 54% yield) as a yellowish solid.

[0357] Step 3. To a solution of tert-butyl N-(6-bromo-2-chloro-3-methoxyphenyl)carbamate (25 g, 74.3 mmol) in dioxane (500 mL) were added trimethylboroxine (50% w / w in THF, 20.51 g, 81.7 mmol), PdCl2(dppf).CH2Cl2 (5.22 g, 7.43 mmol) and aqueous Na2CO3 solution (2 M, 111 mL, 223 mmol). The mixture was heated at 100 °C for 16 h, cooled to room temperature, and then the volatiles were removed under vacuum. EtOAc and water were added. The organic layer was separated, washed with brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 30% EtOAc in heptane to give tert-butyl N-(2-chloro-3-methoxy-6-methylphenyl)carbamate (13.8 g, 68% yield) as a yellowish solid.

[0358] Step 4. Hydrochloric acid in dioxane (4 M, 100 mL) was added to a solution of tert-butyl N-(2-chloro-3-methoxy-6-methylphenyl)carbamate (13.8 g, 50.8 mmol) in MeOH (100 mL). After 3 h, the volatiles were evaporated to dryness under vacuum to give a white solid to which 250 mL of EtOAc and 250 mL of saturated aqueous NaHCO3 were added with vigorous stirring. The organic layer was separated. The aqueous layer was back-extracted with EtOAc. The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 0 - 30% EtOAc in heptane to give 2-chloro-3-methoxy-6-methylaniline (7.9 g, 91% yield) as a clear oil which solidified on standing. 1 1H NMR (400 MHz, chloroform-d) δ 6.95 - 6.79 (m, 1H), 6.27 (dd, J = 8.3, 1.5 Hz, 1H), 4.06 (br s, 2H), 3.83 (d, J = 1.6 Hz, 3H), 2.12 (d, J = 0.8 Hz, 3H). MS: [M + 1]: 172.2.

[0359] Preparation of Aryl Amine A5 The compounds of the present invention can be prepared from arylamine A5 shown in Scheme A5 and prepared as described herein. Commercially available 3-methoxy-2-methyl-aniline can be chlorinated with a chlorinating reagent to produce arylamine A5.

Chemical formula

[0360] Step 1. To a solution of 3-methoxy-2-methyl-aniline (100 g, 729 mmol) in DCM (500 mL) at 0 °C, NCS (98 g, 734 mmol) was added in 4 portions (each addition was made every 15 minutes). 100 g of silica gel was added 30 minutes after the last addition, and the mixture was evaporated under vacuum and purified by silica gel chromatography (dry packing) eluting with a gradient of 0-10% EtOAc in hexane to give 6-chloro-3-methoxy-2-methyl-aniline (55.6 g, 44% yield) as an orange solid. 1 H NMR (400 MHz, chloroform-d) δ 7.08 (d, J = 8.8 Hz, 1H), 6.28 (d, J = 8.8 Hz, 1H), 4.02 (br s, 2H), 3.78 (s, 3H), 2.07 (s, 3H). MS: [M+1]: 172.3.

[0361] Preparation of arylamine A6 The compounds of the present invention can be prepared from arylamine A6 shown in Scheme A6 and prepared as described herein. Commercially available 3-amino-2,4-dichloro-phenol can be protected at O with a suitable protecting group, e.g., O-PMB, to produce arylamine A6.

Chemical formula

[0362] A suspension containing 3-amino-2,4-dichloro-phenol.HCl salt (20 g, 93.3 mmol) in DMF (150 mL) was added with 1-(chloromethyl)-4-methoxy-benzene (14.0 mL, 103 mmol), tetrabutylammonium iodide (1 g, 3.00 mmol) and Cs2CO3 (64.0 g, 196 mmol). The mixture was stirred at 40 °C overnight, then diluted with water, stirred for 20 minutes and filtered. The precipitate was washed with water and dried under vacuum. The obtained crude product was purified by silica gel chromatography eluting with a gradient of 0-100% DCM in hexane to give 2,6-dichloro-3-[(4-methoxyphenyl)methoxy]aniline (20 g, 72% yield) as an off-white solid. 1 1H NMR (400 MHz, chloroform-d) δ 7.38 - 7.30 (m, 2H), 7.05 (d, J = 8.9 Hz, 1H), 6.92 - 6.77 (m, 2H), 6.32 (s, 1H), 5.01 (s, 2H), 4.46 (s, 2H), 3.79 (s, 3H). MS: [M+1]: 298.0.

[0363] Preparation of arylamine A7. The compounds of the present invention are shown in Scheme A7 and can be prepared from the main intermediates A7, A8 or A9 prepared as described herein (adapted from J. AM. CHEM. SOC. 2004, 126, 1150 - 1160). Commercially available 3-methoxyaniline can have its N protected with a suitable protecting group such as NH-PIV. A suitable R 1 , for example CH3 or CD3 can be introduced using a directed orthometalation approach. The NH-PIV protecting group can be cleaved under acidic conditions and the remaining ortho positions to nitrogen can be brominated with a suitable brominating reagent such as NBS. At this point, bromine can be substituted with a boronic acid ester under metal-mediated conditions and further derivatized with a suitable R 2 , for example CH3 or CD3 to produce the main intermediates A7, A8 or A9. Alternatively, the N of bromoaniline can be protected with a suitable protecting group such as NH-Boc before bromo substitution. In this case, NH-Boc can be cleaved under acidic conditions to produce the main intermediates A7, A8 or A9. [Chem.]

[0364] Arylamine A7 Step 1. To a solution containing 3-methoxyaniline (50 g, 406 mmol, 45.5 mL), pyridine (66 mL, 816 mmol), and DMAP (500 mg, 4.1 mmol) in DCM (500 mL), 2,2-dimethylpropanoyl chloride (51 mL, 416 mmol) was slowly added. After 1 hour, 1 N aqueous HCl was added and the layers were separated. The aqueous layer was back-extracted with CH2Cl2. The combined organic layers were washed with 1 N aqueous HCl and brine, then dried over Na2SO4, filtered, and evaporated to dryness to obtain N-(3-methoxyphenyl)-2,2-dimethyl-propanamide (84 g, quantitative yield).

[0365] Step 2. To a solution of N-(3-methoxyphenyl)-2,2-dimethyl-propanamide (82 g, 396 mmol) in THF (820 mL), nBuLi (2.5 M, 325 mL, 813 mmol) was added dropwise at 0 °C. After 2 hours at 0 °C, the solution was cooled to -78 °C and CD3I (27 mL, 434 mmol) was added dropwise. The mixture was stirred at room temperature for 16 hours. The mixture was poured into 1 N aqueous HCl and extracted twice with EtOAc. The combined organic layers were dried over Na2SO4, filtered, and concentrated to dryness to obtain N-[3-methoxy-2-(trideuteromethyl)phenyl]-2,2-dimethylpropanamide (82 g, 92% yield) as a white solid.

[0366] Step 3. N-[3-Methoxy-2-(trideuteriomethyl)phenyl]-2,2-dimethyl-propanamide (81.5 g, 363 mmol) in dioxane (300 mL) and concentrated HCl (12 M, 300 mL) was heated to reflux for 24 h. The dark mixture was cooled to 0 °C in an ice bath, neutralized with 2 N aqueous NaOH, and extracted twice with EtOAc. The combined organic extracts were washed with brine, dried over Na SO , filtered, and concentrated to dryness to give a dark residue that was purified by silica gel chromatography eluting with a gradient of 0 to 50% EtOAc in heptane to give 3-methoxy-2-(trideuteriomethyl)aniline (36 g, 71% yield) as a clear oil.

[0367] Step 4. To a solution of 3-methoxy-2-(trideuteriomethyl)aniline (35 g, 250 mmol) in DCM (500 mL) was added NBS (45 g, 253 mmol) at 0° C. The mixture was stirred at 0° C. for 3 h, concentrated to approximately 75 mL, and filtered. The filtrate was evaporated to dryness, and the residue was purified by silica gel chromatography eluting with a gradient of 0 to 50% EtOAc in heptane to give 6-bromo-3-methoxy-2-(trideuteriomethyl)aniline (35 g, 64% yield).

[0368] Step 5. To a solution of 6-bromo-3-methoxy-2-(trideuteriomethyl)aniline (35 g, 160 mmol), DMAP (3.90 g, 32 mmol), and DIPEA (415 mmol, 72.3 mL) in THF (500 mL) was added tert-butoxycarbonyl tert-butyl carbonate (87.2 g, 400 mmol). The mixture was heated to reflux for 18 h. The volatiles were removed under vacuum, and the residue was filtered through silica gel eluting with 50% EtOAc in heptane to give a mixture of tert-butyl N-[6-bromo-3-methoxy-2-(trideuteriomethyl)phenyl]carbamate and tert-butyl N-[6-bromo-3-methoxy-2-(trideuteriomethyl)phenyl]-N-tert-butoxycarbonyl-carbamate (64 g) as a clear oil, which was dissolved in methanol (500 mL). K2CO3 (110 g, 796 mmol) was added, and the mixture was stirred at 60 °C for 48 h. The volatiles were removed under vacuum. EtOAc and water were added to the residue. The organic layer was separated, washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography eluting with a gradient of 0-40% EtOAc to give tert-butyl N-[6-bromo-3-methoxy-2-trideuteriomethyl)phenyl]carbamate (50 g, quantitative yield) as a clear oil.

[0369] Step 6. To a solution of tert-butyl N-[6-bromo-3-methoxy-2-(trideuteromethyl)phenyl]carbamate (33 g, 103 mmol) in dioxane (700 mL) were added bis(pinacolato)diboron (51 g, 201 mmol), KOAc (35.5 g, 362 mmol), and PdCl2(dppf).CH2Cl2 (7.6 g, 10.4 mmol). The mixture was degassed under vacuum, filled with nitrogen again, and stirred under reflux for 18 h. The mixture was cooled to room temperature and concentrated to a smaller volume. The black residue was diluted with EtOAc and eluted with 2 L of heptane containing 50% EtOAc by filtration through a silica gel pad (250 g). The filtrate was evaporated, and the residue was purified by silica gel chromatography eluting with a gradient of 0-20% EtOAc in heptane to give tert-butyl N-[3-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trideuteromethyl)phenyl]carbamate (22.5 g, 59% yield), which solidified upon standing under vacuum.

[0370] Step 7. To a solution of PdCl2(dppf).CH2Cl2 (5.3 g, 7.24 mmol) in DMF (500 mL) were successively and rapidly added CD3I (60.6 g, 418 mmol, 26 mL), tert-butyl N-[3-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trideuteromethyl)phenyl]carbamate (52 g, 142 mmol), and aqueous tribasic potassium phosphate (2 M, 350 mL). Nitrogen was bubbled through the solution for 2 min, then the mixture was stirred at 80 °C for 30 min under a nitrogen atmosphere, then cooled to room temperature, and EtOAc was added. The organic layer was washed with water, brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with a gradient of 0-20% EtOAc in heptane to give tert-butyl N-[3-methoxy-2,6-bis(trideuteromethyl)phenyl]carbamate (15 g, 41% yield) as a viscous clear oil.

[0371] Step 8. To a solution of tert-butyl N-[3-methoxy-2,6-bis(trideuteriomethyl)phenyl]carbamate (22.5 g, 87.4 mmol) in MeOH (100 mL) was added HCl in dioxane (4 M, 100 mL). After 3 h, the volatiles were removed in vacuo to give a white solid. EtOAc and water were added, followed by saturated aqueous NaHCO3 until a basic pH was achieved. The organic layer was washed with brine, dried over Na2SO4, filtered, and evaporated to dryness. The residue was purified by silica gel chromatography eluting with a gradient of 0 to 40% EtOAc in heptane to give 3-methoxy-2,6-bis(trideuteriomethyl)aniline (7.5 g, 55% yield) as a clear oil. 1 H NMR (400 MHz, chloroform-d) δ 6.96 (dd, J = 8.3, 2.6 Hz, 1H), 6.38 (dd, J = 8.3, 2.5 Hz, 1H), 3.86 (d, J = 2.4 Hz, 3H), 3.64 (s, 2H). MS: [M+1]: 158.3.

[0372] Alternative route used to prepare arylamine A8 (without Boc) Step 1. 6-Bromo-3-methoxy-2-methylaniline (3.4 g, 15.7 mmol), bis(pinacolato)diboron (5.58 g, 21.98 mmol), and CsCO (15.4 g, 47.1 mmol) were placed in anhydrous 1,4-dioxane (68 mL) in a sealed tube, and the reaction mixture was purged with nitrogen gas for 15 minutes. Then, PdCl(dppf) (1.92 g, 2.36 mmol) was added to the reaction mixture, which was then heated at 100 °C for 2 hours. After completion, the reaction mixture was quenched with ice water and extracted with EtOAc (3 × 100 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated to give the crude product, which was purified by silica gel chromatography eluting with a gradient of 10-12% EtOAc in hexanes to give 3-methoxy-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (2.50 g, 60% yield).

[0373] Step 2. 3-Methoxy-2-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)aniline (2.5 g, 9.39 mmol), iodomethane-d3 (4.08 g, 28.19 mmol), and tribasic potassium phosphate (9.95 g, 46.9 mmol) were taken in anhydrous DMF (50 mL) in a sealed tube, and the reaction mixture was purged with nitrogen for 15 minutes. PdCl2(dppf) (0.766 g, 0.939 mmol) was then added, and the reaction mixture was heated at 80 °C for 2 hours. Upon completion, the reaction mixture was quenched using ice water and extracted using EtOAc (3 × 50 mL). The combined organic layers were dried over Na2SO4, filtered, and concentrated to give the crude product, which was purified by silica gel chromatography eluting with a gradient of 6–8% EtOAc in hexanes to give pure 3-methoxy-2-methyl-6-(methyl-d3)aniline as a colorless liquid (0.75 g, 51% yield). 1 H NMR (400MHz, DMSO-d6) δ6.68 (d, J = 7.2 Hz, 1H), 6.31 (d, J = 7.4 Hz, 1H), 4.63 (s, 2H), 3.55 (s, 3H), 1.94 (s, 3H). MS:[M+1]:155.3.

[0374] Preparation of arylamine A10 The compounds of the present invention can be prepared from key intermediate A10, which is shown in Scheme A8 and can be prepared as described herein. Commercially available 3-methoxy-2-nitrobenzoic acid can be brominated. The acid can then be esterified, the bromo can be converted to methyl, and the nitro can be reduced to amino. The resulting amino can be converted to bromo under Sandmeyer conditions, and the ester can then be saponified. The resulting acid can then be converted to NHBoc under Curtius conditions. In this case, the NH-Boc can be cleaved under acidic conditions to produce key intermediate A10. [ka]

[0375] Step 1. To 3-methoxy-2-nitro-benzoic acid (10.04 g, 50.93 mmol) and AgSO (8.10 g, 26.0 mmol) in the dark, concentrated sulfuric acid (200 mL) and molecular bromine (9.4 g, 58.6 mmol, 3.0 mL) were added dropwise. The mixture was stirred in the dark for 3.5 h, then quenched by the addition of crushed ice, cooled in an ice bath, and stirred. The solid was collected by filtration, washed with HO, and air-dried. The resulting solid was taken up in acetone (300 mL), filtered, and the residue (silver salt) was washed with acetone. The filtrate was dried over MgSO, filtered, and concentrated to give 6-bromo-3-methoxy-2-nitro-benzoic acid (14.64 g, 100% yield) as a purple solid.

[0376] Step 2. To a solution of 6-bromo-3-methoxy-2-nitro-benzoic acid (14.64 g, 53.0 mmol) in DMF (140 mL) was added anhydrous potassium carbonate (14.66 g, 106.1 mmol), followed by methyl iodide (11.4 g, 80.3 mmol, 5.0 mL). The reaction mixture was stirred for 2 hours, after which HO was added dropwise (420 mL). The solid was collected by filtration, washed with HO, air-dried, and then dried under vacuum to give methyl 6-bromo-3-methoxy-2-nitro-benzoate (12.89 g, 84% yield) as a pale yellow-brown solid.

[0377] Step 3. A solution containing methyl 6-bromo-3-methoxy-2-nitro-benzoate (6.0 g, 20.7 mmol) in dioxane (100 mL) and aqueous Na2CO3 solution (2 M, 31 mL, 62.3 mmol) was bubbled with N2, and then Pd(dppf)Cl2 (1.64 g, 2.01 mmol) and trimethylboroxin (6.74 g, 26.8 mmol, 7.5 mL) were added. The solution was bubbled with N2. The lid of the vessel was closed and the mixture was stirred at 100 °C overnight. The reaction mixture was cooled to room temperature, poured into H2O, and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over Na2SO4, filtered through a silica plug, and concentrated under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of EtOAc (0 - 100%) in Hep. Appropriate fractions were combined and concentrated under vacuum to give methyl 3-methoxy-6-methyl-2-nitro-benzoate (3.06 g, 66% yield) as a pale yellowish brown waxy solid.

[0378] Step 4. To a solution of methyl 3-methoxy-6-methyl-2-nitro-benzoate (3.06 g, 13.6 mmol) in MeOH (225 mL) was added palladium carbon (10% w / w, 1.42 g, 1.33 mmol) slurried in some of the MeOH. H2 was bubbled through the mixture and the mixture was stirred under a hydrogen atmosphere for 2 h. The suspension was filtered through celite and the filtrate was concentrated and then dried under vacuum to give methyl 2-amino-3-methoxy-6-methyl-benzoate (2.56 g, 97% yield) as a pale amber oil.

[0379] Procedure 5. To a solution of methyl 2-amino-3-methoxy-6-methyl-benzoate (2.13 g, 10.9 mmol) in DMF (12 mL) and MeCN (18 mL) was added tert-butyl nitrite (2.0 mL, 17 mmol), followed by the addition of copper(II) bromide (2.86 g, 12.8 mmol). The reaction mixture was stirred at 55 °C for 9 minutes, then cooled to room temperature, diluted with H2O, and extracted with EtOAc (3×). The combined organic extracts were washed with saturated aqueous NH4Cl and brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (dry packing) eluting with a gradient of EtOAc in Hep (0 - 70%) to give methyl 2-bromo-3-methoxy-6-methyl-benzoate (1.52 g, 54% yield) as a yellow oil. 1 1H NMR (400 MHz, chloroform-d) δ 7.15 - 7.05 (m, 1H), 6.84 (d, J = 8.4 Hz, 1H), 3.95 (s, 3H), 3.88 (s, 3H), 2.26 (d, J = 0.7 Hz, 3H).

[0380] Procedure 6. To a solution of methyl 2-bromo-3-methoxy-6-methyl-benzoate (1.52 g, 5.87 mmol) in MeOH (15 mL) and THF (15 mL) were added aqueous NaOH solution (4 M, 15 mL, 60.0 mmol) and 30% hydrogen peroxide solution (1.5 mL). The reaction mixture was stirred at 70 °C overnight, then stirred at 90 °C for 4 days. The reaction mixture was cooled to room temperature and the volatile materials were removed under vacuum. The residue was diluted with 3N HCl (20 mL) and extracted with CHCl3 / iPrOH (4:1, 4×). The combined organic extracts were concentrated and then dried under vacuum. The crude product was purified by silica gel chromatography (dry packing) eluting with a gradient of MeOH in CH2Cl2 (0 - 10%) using 1% AcOH as the eluent to give 2-bromo-3-methoxy-6-methyl-benzoic acid (896 mg, 62% yield) as a white solid.

[0381] Step 7. To a solution containing 2-bromo-3-methoxy-6-methyl-benzoic acid (1.15 g, 4.68 mmol), triethylamine (1.42 g, 14.1 mmol, 2.0 mL), and tert-butanol (1.73 g, 23.4 mmol, 2.25 mL) in toluene (8 mL) was added DPPA (1.93 g, 7.01 mmol, 1.52 mL). The mixture was heated to reflux for 1 hour and cooled to room temperature. Volatiles were removed under vacuum. The residue was diluted with 15% aqueous citric acid and extracted with EtOAc (3×). The combined organic layers were washed with 1N aqueous NaOH, brine, dried over Na2SO4, filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (dry packing) eluting with a gradient of EtOAc (0 - 30%) in Hep. Appropriate fractions were combined and concentrated under vacuum to give tert-butyl N-(2-bromo-3-methoxy-6-methyl-phenyl)carbamate (1.34 g, 91% yield) as a colorless oil.

[0382] Step 8. To a solution of tert-butyl N-(2-bromo-3-methoxy-6-methyl-phenyl)carbamate (1.34 g, 4.24 mmol) in MeOH (10 mL) was added dioxane containing HCl (4 M, 10.5 mL, 42.0 mmol). The reaction mixture was stirred at room temperature for 75 minutes. The solution was evaporated to dryness under vacuum, then suspended in saturated aqueous NaHCO3 and extracted with DCM (3×, phase separator). The combined organic extracts were concentrated and the crude product was purified by silica gel chromatography eluting with a gradient of EtOAc (0 - 50%) in Hep. Appropriate fractions were combined and concentrated under vacuum to give 2-bromo-3-methoxy-6-methyl-aniline (807 mg, 88% yield) as an off-white waxy solid. MS: [M+1]: 218.0.

[0383] Preparation of Aryl Amine A11 The compounds of the present invention can be prepared from the main intermediate A11 shown in Scheme A9 and prepared as described herein. Commercially available 6-bromo-3-methoxy-2-methylbenzoic acid can be converted to N-Boc under Curtius rearrangement conditions. NH-Boc can be cleaved under acidic conditions to produce the main intermediate A11.

Chemical formula

[0384] Step 1. To a solution of 6-bromo-3-methoxy-2-methyl-benzoic acid (1 g, 4.08 mmol), triethylamine (1.23 g, 12.2 mmol, 1.70 mL), and tert-butanol (1.55 g, 20.9 mmol) in toluene (7 mL) was added [azido(phenoxy)phosphoryl]oxybenzene (1.72 g, 6.25 mmol, 1.35 mL). The mixture was heated to reflux for 5 hours and cooled to room temperature. Volatiles were removed under vacuum. The residue was diluted with 15% aqueous citric acid and extracted with EtOAc (3×). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel chromatography (dry packing) eluting with a gradient of EtOAc in Hep (0 - 30%). Appropriate fractions were combined and concentrated under vacuum to give tert-butyl N-(6-bromo-3-methoxy-2-methyl-phenyl)carbamate (1.41 g, quantitative yield) as a colorless oil, which was used without further purification in the next step.

[0385] Step 2. To a solution of tert-butyl N-(6-bromo-3-methoxy-2-methyl-phenyl)carbamate (1.41 g, 4.46 mmol) in MeOH (22 mL) was added HCl in dioxane (4 M, 22 mL). The reaction mixture was stirred at room temperature for 80 min. The solution was evaporated to dryness in vacuo, then suspended in saturated aqueous NaHCO3 and extracted with DCM (3x). The combined organic extracts were dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of EtOAc (0-50%) in Hep. The appropriate fractions were combined and concentrated in vacuo to give 6-bromo-3-methoxy-2-methyl-aniline (586 mg, 61% yield) as a white solid. 1 H NMR (400 MHz, chloroform-d) δ 7.23 (dd, J = 8.8, 0.6 Hz, 1H), 6.26 (d, J = 8.8 Hz, 1H), 4.06 (br s, 2H), 3.78 (s, 3H), 2.09 (t, J = 0.5 Hz, 3H). MS: [M+1]: 218.0.

[0386] Chiral separation of selected compounds The racemic mixture of atropisomers was separated using chiral SFC on a Mettler Toledo Minigram SFC (MTM), a Waters Prep 15 SFC-MS (WP15), or a Waters Prep 100 SFC-MS (WP100) (Table 3). The appropriate column was selected to achieve satisfactory peak resolution. Appropriate fractions for each peak were combined, concentrated, taken up in a mixture of water and a suitable water-soluble organic solvent, such as EtOH, IPA, CHCN, or a mixture thereof, and lyophilized. The separated products were reanalyzed by chiral SFC to assess chiral purity.

[0387] C1A is Phenomenex Lux Cellulose-2, 10×250mm, 5μm; C1B is Phenomenex Lux Cellulose-2, 30×250mm, 5μm; C2 is Chiral Technologies IA, 10×250mm, 5μm; C3 is Chiral Technologies IC, 10×250mm, 5μm; C4 is Chiral Technologies ID, 10×250mm, 5μm; C5 is Chiral Technologies IG, 10×250mm, 5μm; C6 is Chiral Technologies AS, 10×250mm, 5μm; C7 is Phenomenex Lux Cellulose-4, 10×250mm, 5μm.

[0388] The structural assignment of the separated atropisomers was supported by biological activity, and was assigned such that the biologically active enantiomer had the (S) configuration, and its confirmation was performed by X-ray crystal structure analysis of the major compound.

Table 3-1

Table 3-2

[0389] Example 2. Enzymatic assay For the detection of Myt1 kinase activity, a recombinant human Myt1 kinase assay that measures the hydrolysis of ATP using a commercially available ADP-Glo assay (ADP-Glo™ Kinase Assay from Promega, 10000 assays, #V9102) was utilized. Briefly, 5 μL aliquots of recombinant human Myt1 (full-length PKMYT1 recombinant human protein expressed in insect cells #A33387 from Thermo Fisher, purity approximately 80%) were added to reaction buffer (70 mM HEPES, 3 mM MgCl 2、 3 mM MnCl 2、The Myt1 enzyme solution (50 μg / ml PEG20000, 3 μM sodium orthovanadate, 1.2 mM DTT) was prepared and added to a 384-well white polystyrene flat-bottom, non-treated microplate (Corning #3572). Five μL of compound (diluted in reaction buffer to 0.5% DMSO) was then added to the microplate, and the plate was briefly spun and incubated at 22°C for 15 minutes. Ultra-Pure adenosine triphosphate (ATP) solution (ADP-Glo kit from Promega) was diluted in reaction buffer, and 5 μL was added to the microplate, spun briefly, and incubated at 30°C for 60 minutes. The final Myt1 enzyme concentration was 18 nM, and the final ATP concentration was 10 μM. After the 60-minute incubation, 15 μL of ADP-Glo reagent was added, and the plate was briefly spun, sealed, and incubated in the dark for 40 minutes at 22°C. Following this, 30 μL of kinase detection reagent was added per well, the plate was briefly spun, sealed, and incubated in the dark for 45-60 minutes at 22°C. Luminescence was read using Envision (250 ms integration). IC values were calculated for each inhibitor compound tested. 50 and % maximum inhibition was calculated.

[0390] Exemplary prepared compounds and their activities are shown in Table 4 below. [Table 4-1] [Table 4-2] [Table 4-3] [Table 4-4] [Table 4-5]

Table 4-6

Table 4-7

Table 4-8

Table 4-9

Table 4-10

[0391] Example 3. Genetic Verification Two sgRNAs for PKMYT1 and one sgRNA for LacZ (control) were transduced into RPE1-hTERT Cas9 TP53− / − parental (WT) and CCNE1 overexpressing clones. Infected cells were seeded at low density and their ability to form colonies of less than 50 cells was measured. After growing for 10 days, colonies were stained, imaged, and quantified. Using the colony formation survival assay, the inventors observed significant cell maladaptation in CCNE1 overexpressing cells compared to parental cells transduced with PKMYT1 sgRNA (Figures 3A and 3B). This experiment was repeated using FT282-hTERT TP53− / − parental (WT) and CCNE1 overexpressing clones, and the same results were observed (Figures 4A and 4B).

[0392] To determine whether the kinase activity of PKMYT1 governed the maintenance of CCNE1-overexpressing RPE1-hTERT Cas9 TP53- / - cells, the PKMYT1 open reading frame (ORF) was cloned into an inducible mammalian expression vector. Subsequently, sgRNA-resistant silent mutations were created within the PKMYT1 ORF sequence by PCR mutagenesis. A single point mutation resulting in an amino acid change from asparagine (N) to alanine (A) at residue 238 was generated. The N238A amino acid change in the kinase domain resulted in a catalytically inactive PKMYT1 mutant. Stable cell lines containing either the wild-type PKMYT1 ORF or the kinase-dead N238A mutant were generated in the RPE1-hTERT Cas9 TP53- / - parental and CCNE1-overexpressing clones (Figure 5A). These stable cell lines were transduced with either a LacZ nonspecific sgRNA or PKMYT1 sgRNA #4. Cells were then seeded at low density to measure their ability to form colonies of >50 cells. After 10 days of growth, colonies were stained, imaged, and quantified. Expression of the sgRNA-resistant PKMYT1 ORF, but not the catalytically inactive form, rescued the fitness defect induced by transduction of sgRNA #4 in both CCNE1-overexpressing clones (Figures 5B and 5C). These results demonstrate that targeting the kinase activity of PKMYT1 selectively kills CCNE1-overexpressing cells.

[0393] Example 4. Pharmacological validation RPE1-hTERT Cas9 TP53- / - parental (WT) and CCNE1-overexpressing clones were treated with Compound 133 in a dose titration assay, and cell viability was determined. CCNE1-overexpressing cells were found to be more sensitive to Compound 133 compared to corresponding WT cells (Figure 3C). FT282-hTERT TP53 R175HSimilar effects were observed in WT and CCNE1 overexpression clones (Figure 4C). For the dosing response proliferation assays using the RPE1-hTERT and FT282-hTERT cell lines, cells were seeded in 96-well plates and dosed with serially diluted Myt1 inhibitor. Cells were imaged once a day using an IncuCyte S3 microscope, and the well confluence percentage was calculated over time. The experiment was terminated when the cells reached the fourth population doubling, and IC 50 curves were plotted. The confluence percentage was calculated relative to the cell confluence in the untreated wells.

[0394] For a panel of 16 cancer cell lines where the level of CCNE1 was either normal (n = 8) or elevated (n = 8), their sensitivity to compound 28 was evaluated in a cell proliferation assay (Figure 6). In these cancer cell line proliferation assays, dose-response curves were generated as follows. Cells were seeded in 96-well plates and dosed with serially diluted compound 28. After 7 days, the growth status of these cells was evaluated using Cell Titer Glo (CTG), and IC 50 values were plotted.

[0395] A similar experiment was conducted in a panel of 8 cancer cell lines with either wild-type FBXW7 (n = 5) or FBXW7 mutation (n = 3) to evaluate their sensitivity to compound 95 in a cell proliferation assay (Figure 7). In these cancer cell line proliferation assays, dose-response curves were generated as follows. Cells were seeded in 96-well plates and dosed with serially diluted Myt1 inhibitor. Cells were imaged once a day using an IncuCyte S3 microscope, and the well confluence percentage was calculated over time. The experiment was terminated when the cells reached the fourth population doubling, and IC 50 curves were plotted. The confluence percentage was calculated relative to the cell confluence in the untreated wells.

[0396] Other embodiments Various changes and modifications of the described invention will be apparent to those skilled in the art without departing from the scope and spirit of the invention. Although the invention has been described in connection with specific embodiments, it is to be understood that the claimed invention should not be unduly limited to such specific embodiments. Indeed, various modifications of the embodiments of the invention that are apparent to those skilled in the art are intended to be within the scope of the invention.

[0397] Other embodiments are within the scope of the claims.

Claims

1. A compound of formula (I): 【Chemical 1】 〔wherein, Each of X, Y and Z is independently N or CR ２ ; R １ is hydrogen, optionally substituted C １－６ alkyl, optionally substituted C ２－６ alkenyl, optionally substituted C ２－６ alkynyl, optionally substituted C ３－８ cycloalkyl, optionally substituted C ３－８ cycloalkenyl, optionally substituted C ２－９ heterocyclyl, optionally substituted C ２－９ heterocyclyl C １－６ alkyl, optionally substituted C ６－１０ aryl, optionally substituted C １－９ heteroaryl, optionally substituted C １－９ heteroaryl C １－６ alkyl, halogen, cyano, -N(R ７ ) ２ , -OR ７ , -C(O)N(R ８ ) ２ , -SO ２ R ７Ａ , or -Q-R ７Ｂ ; each R2 is independently hydrogen, optionally substituted C1-6 alkyl, C2-6 alkenyl, optionally substituted C2-6 alkynyl, optionally substituted C6-10 aryl, optionally substituted C1-9 heteroaryl, halogen, -OR7, or -Q-R7B; or R １ is R １ and is vicinal to one R ２ and combines with C ３－６ to form alkylene; R ３ and R ４ each of which is independently optionally substituted C １－６ alkyl or halogen; R ５ is H or -NH₂; R ６ is -C(O)NH₂, -C(O)CH₃, or -SO ２ Me; Each R ７ is independently hydrogen, optionally substituted C １－６ alkyl, optionally substituted C ６－１０ aryl C １－６ alkyl, optionally substituted C ３－８ cycloalkyl, optionally substituted C ６－１０ aryl, optionally substituted C ２－９ heterocyclyl, optionally substituted C １－９ heteroaryl, optionally substituted C １－９ heteroaryl C １－６ alkyl, or -SO ２ R ７Ａ or; or two R ７ groups, together with the atom to which both are attached, combine to form an optionally substituted C ２－９ heterocyclyl; Each R ７Ａ is independently C １－６ alkyl, C ３－８ cycloalkyl, or C ６－１０ aryl; Each R ７Ｂ are independently hydroxyl, optionally substituted C １－６ Alkyl, optionally substituted C ６－１０ Aryl, optionally substituted C ２－９ Heterocyclyl, optionally substituted C １－９ Heteroaryl, —N(R ７ ) ２ , -C(O)N(R ８ ) ２ , -SO ２ R ７Ａ or optionally substituted alkoxy; Each R ８ is independently hydrogen, optionally substituted C １－６ alkyl, optionally substituted C ６－１０ aryl, or optionally substituted C １－９ heteroaryl; Q is optionally substituted C １－６ alkylene, optionally substituted C ２－６ alkenylene, optionally substituted C ２－６ alkynylene, optionally substituted C ３－８ cycloalkylene, optionally substituted C ３－８ cycloalkenylene, optionally substituted C ６－１０ arylene, optionally substituted C ２－９ heterocyclylene, or optionally substituted C １－９ heteroarylene), or a pharmaceutically acceptable salt thereof.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is enriched in the atropisomer of formula (IA): [Chemical 2]

3.

4. X is CR ２ The compound according to claim 1 or claim 2, or a pharmaceutically acceptable salt thereof, wherein X is CR. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound has the formula (II):

5. [Chemical 3] The compound according to claim 4, or a pharmaceutically acceptable salt thereof, wherein the compound is enriched in the atropisomer of formula (IIA):

6. The compound according to claim 6, or a pharmaceutically acceptable salt thereof, wherein the compound has the formula (III): 【Chemical Formula 4】 〔wherein,

7. The compound according to claim 6, or a pharmaceutically acceptable salt thereof, wherein the compound is enriched in the atropisomer of formula (IIIA): [Chemical Formula 5]

8. R ２Ａ is hydrogen, optionally substituted C １－６ alkyl, C ２－６ alkenyl, optionally substituted C ２－６ alkynyl, optionally substituted C ６－１０ aryl, optionally substituted C １－９ heteroaryl, halogen, -OR ７ , or -Q-R ７Ｂ , the compound according to claim 1, or a pharmaceutically acceptable salt thereof. The compound according to claim 6 or claim 7, or a pharmaceutically acceptable salt thereof.

9. [Chemical Formula 6]

10.

11. R ２Ａ is hydrogen, optionally substituted C １－６ alkyl, or halogen,

12.

13. R ３ is optionally substituted C １－６ alkyl, a compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof. The compound according to claim 10 or claim 12, wherein the halogen is chlorine. R ３ The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein R is halogen.

14. R ４ is optionally substituted C １－６ alkyl, a compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof.

15. R ４ The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, wherein R is halogen.

16.

17.

18. R ２ The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R is hydrogen.

19. R ２ is optionally substituted C １－６ alkyl, a compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof.

20. R ２ The compound according to claim 15, or a pharmaceutically acceptable salt thereof, wherein R is methyl optionally substituted or isopropyl optionally substituted. The compound according to claim 19, or a pharmaceutically acceptable salt thereof. R ２ The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R is a halogen.

21. R １ The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein R is hydrogen.

22. R １ The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein R is halogen.

23. R １ wherein R is chlorine or bromine

24.

25. R １ is optionally substituted C １－６ alkyl, a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof.

26. R １ The compound according to claim 21, or a pharmaceutically acceptable salt thereof, wherein R is methyl optionally substituted, ethyl optionally substituted, isopropyl optionally substituted, or butyl optionally substituted.

27. R １ is optionally substituted C １－９ The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein the heteroaryl is heteroaryl.

28. R １ is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidinyl, pyrazinyl, pyridazinyl or pyrazolyl, and R １ is optionally substituted with a substituent defined for the optionally substituted C １－９ heteroaryl, the compound according to claim 23, or a pharmaceutically acceptable salt thereof.

29. R １ is optionally substituted C ３－８ The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein is cycloalkyl.

30. R １ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and R １ is optionally substituted with a substituent defined for the optionally substituted C ３－８ cycloalkyl, the compound according to claim 25, or a pharmaceutically acceptable salt thereof.

31. R １ is optionally substituted C ２－９ The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein is a heterocyclyl.

32. R １ is 1,2,3,6 - tetrahydropyridinyl, piperidinyl, morpholinyl, piperazinyl, thiomorpholinyl, oxa - aza - spiro[3,3]heptane, or oxa - aza - bicyclo[3.2.1]octane, and R １ is optionally substituted with a substituent as defined for the optionally substituted C ２－９ heterocyclyl, the compound according to claim 27, or a pharmaceutically acceptable salt thereof.

33. R １ The compound according to claim 25, or a pharmaceutically acceptable salt thereof, wherein R is optionally substituted cyclohexenyl or optionally substituted cyclopentenyl.

34. R １ is optionally substituted C ６－１０ aryl, a compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof.

35. R １ The compound according to claim 30, or a pharmaceutically acceptable salt thereof, wherein R is phenyl optionally substituted.

36. R １ is -Q-R ７Ｂ The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, wherein

37. Q is optionally substituted C ２－６ The compound according to claim 32, or a pharmaceutically acceptable salt thereof, wherein ２－６ is alkynylene.

38. Q is optionally substituted C １－６ The compound according to claim 32, or a pharmaceutically acceptable salt thereof, wherein １－６ is alkylene. n is 0 or 1, Q is optionally substituted C ６－１０ The compound according to claim 32, or a pharmaceutically acceptable salt thereof, wherein Q is an arylene. ６－１０ The compound according to any one of claims 1 to 37, or a pharmaceutically acceptable salt thereof. R ７Ｂ is optionally substituted C ２－９ The compound according to any one of claims 32 to 35, or a pharmaceutically acceptable salt thereof, wherein is a heterocyclyl.

39. R ７Ｂ is optionally substituted C ６－１０ aryl, a compound according to any one of claims 32 to 35, or a pharmaceutically acceptable salt thereof.

40. R １ is methyl, difluoromethyl, trifluoromethyl, fluorine, chlorine, bromine, amino, hydroxyl, cyano, oxo, -C(O)NH ２ , -C(O)NH(Me), -C(O)N(Me) ２ , -(CH ２ ), ｎ -C(O)OH, and -(CH ２ ), ｎ -C(O)Ot-Bu, and is optionally substituted with one, two or three groups independently selected from the group consisting of

41.

42.

43. R １ is -N(R ７ ) ２ The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof.

44. R １ The compound according to claim 39, or a pharmaceutically acceptable salt thereof, wherein R is diethylamino.

45. A compound selected from the group consisting of the following compounds, or a pharmaceutically acceptable salt thereof. R ５ The compound according to any one of claims 1 to 40, or a pharmaceutically acceptable salt thereof, wherein R is hydrogen.

46. R ５ is -NH ２ The compound according to any one of claims 1 to 41, or a pharmaceutically acceptable salt thereof, wherein The compound according to claim 45, wherein the compound is R ６ is -C(O)NH ２ The compound according to any one of claims 1 to 42, or a pharmaceutically acceptable salt thereof, wherein R is -C(O)NH.

47. R ６ is -SO ２ Me, the compound according to any one of claims 1 to 43, or a pharmaceutically acceptable salt thereof. The compound according to any one of claims 1 to 46 or a pharmaceutically acceptable salt thereof, and 【Table 1-1】 【Table 1-2】 【Table 1-3】 【Table 1-4】 【Table 1-5】 【Table 1-6】 【Table 1-7】 【Table 1-8】 【Table 1-9】 【Table 1-10】 【Table 1-11】 【Table 1-12】 【Table 1-13】 【Table 1-14】 【Table 1-15】 【Table 1-16】 【Table 1-17】 【Table 1-18】 【Table 1-19】 【Table 1-20】 【Table 1-21】 【Table 1-22】 【Table 1-23】 【Table 1-24】 【Table 1-25】 【Table 1-26】 【Table 1-27】 【Table 1-28】 【Table 1-29】 ​ ​ 【Chemical Formula 7】 ​ ​ ​ A pharmaceutical composition comprising a pharmaceutically acceptable excipient thereof. [

48. ] The pharmaceutical composition according to claim 47, wherein the compound is isotope-enriched with deuterium. [

49. ] The compound is [Chemical Formula 8] or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition according to claim 48.

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