Compounds, pharmaceutical compositions, and methods of preparing and using compounds

Abstract

To provide compounds and pharmaceutically acceptable salts thereof that may be used in the treatment of subjects in need thereof.SOLUTION: The compounds disclosed herein may be inhibitors of tyrosine and threonine-specific cdc2-inhibitory kinase (Myt1). Also disclosed are pharmaceutical compositions containing the compounds or pharmaceutically acceptable salts thereof, and methods of their preparation and use.SELECTED DRAWING: Figure 1A

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, particularly diseases or conditions that depend on the activity of the membrane-bound tyrosine- and threonine-specific cdc2 inhibitory kinase (Myt1) (gene name PKMYT1) (e.g., cancers harboring CCNE1 amplification / overexpression or FBXW7-mutated cancers). [Background technology]

[0002] DNA is continuously exposed to both endogenous (e.g., stalled replication forks, reactive oxygen species) and exogenous (UV, ionizing radiation, chemicals) damaging factors that can lead to DNA damage. As a result, cells have established sophisticated mechanisms to counter these harmful events, which could 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 various phases of the cell cycle, including the G1, S, G2, and mitotic checkpoints. The majority of cancer cells lack their G1 checkpoint due to p53 mutations and therefore rely on the G2 checkpoint to correct the required DNA damage before progressing into mitosis and dividing into two daughter cells.

[0003] There is a need for new anti-cancer therapeutic approaches, such as those that utilize small molecules, particularly therapies that allow for directed cancer treatment. Summary of the Invention

[0004] In one aspect, the present invention provides a compound of formula (I): [ka] [During the ceremony, Each of X, Y, and Z is independently N or CR2 and; R 1 and each R 2 are independently 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, -SO2N(R 8 )2, -SO2R 7A , or -QR 7B or R 1 But R 1 One R that is vicinal to 2 in combination with an optionally substituted C 3-6 forming an alkylene; R 3 and R 4 each independently represents an optionally substituted C 1-6 is alkyl or halogen; R 5 is H or -N(R 7 )2; R 6 is -C(O)NH(R 8 ), -C(O)R 7A , or -SO2R 7A and; Each R 7 are independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl C1-6 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 -SO2R 7A or two R 7 The group, together with the atoms attached to both, forms an optionally substituted C 2-9 forming a heterocyclyl; Each R 7A are independently optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, or optionally substituted C 6-10 is 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, -SO2N(R 8 )2, -SO2R 7A or optionally substituted alkoxy; Each R 8 are independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 Aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 8are combined with the atoms attached to them to form an optionally substituted C 2-9 forming a heterocyclyl; Q is an 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.

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

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

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

[0008] In some embodiments, the compound has 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(R 8 )2, -SO2R 7A , or -QR 7B In some embodiments, the compound has an atropisomer of formula (IIIA): [ka] It is enriched for

[0009] In some embodiments, R 2A is hydrogen, optionally substituted C 1-6 alkyl, or halogen.

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

[0011] In some embodiments, R 2 is hydrogen. In some embodiments, R 2 is an optionally substituted C 1-6 It is alkyl. R 2 is optionally substituted methyl or optionally substituted isopropyl. 2 is a 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 an optionally substituted C 1-6 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 an optionally substituted C 1-9 In some embodiments, R is heteroaryl. 1 is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl or pyrazolyl, and R 1 is an optionally substituted C 1-9 Optionally substituted with the substituents defined for heteroaryl. In some embodiments, R 1 is an optionally substituted C 3-8 In some embodiments, R 1 is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, and R 1 is an optionally substituted C 3-8Optionally substituted with the substituents defined for cycloalkyl. In some embodiments, R 1 is an optionally substituted C 2-9 In some embodiments, R is heterocyclyl. 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; R 1 is an optionally substituted C 2-9 Optionally substituted with the substituents defined for heterocyclyl. In some embodiments, R 1 is an optionally substituted C 3-8 In some embodiments, R 1 is optionally substituted cyclohexenyl or optionally substituted cyclopentenyl. In some embodiments, R 1 is an optionally substituted C 6-10 In some embodiments, R 1 is an optionally substituted phenyl.

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

[0014] In some embodiments, R 1is 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 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 -NH. In some embodiments, R 6 is -C(O)NH(R 8 In some embodiments, R 6 is —C(O)NH. 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, the compound is selected from the group consisting of compounds 1-328 (eg, 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 a cell in which Myt1 is expressed, comprising contacting the cell with a compound disclosed herein.

[0020] In some embodiments, the cell overexpresses CCNE1. In some embodiments, the cell is in a subject.

[0021] In yet another aspect, the present invention provides a method of treating a subject in need thereof, 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 suffers from a disease or condition that has symptoms of cell hyperproliferation and needs 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 for treating cancer in a subject, comprising administering a therapeutically effective amount of a Myt1 inhibitor to a subject in need thereof, wherein the cancer has previously been identified as a cancer in which CCNE1 is overexpressed.

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

[0025] In yet another aspect, the present invention provides a method for 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 cell is 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 for treating cancer in a subject, comprising administering a therapeutically effective amount of a Myt1 inhibitor to a subject in need thereof, wherein the cancer has previously been identified as a cancer having an inactivating mutation in the FBXW7 gene.

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

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

[0030] In some embodiments, the cell is in a subject. The cancer is uterine cancer, colon 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] Abbreviation Abbreviations and terms commonly used in the fields of organic chemistry, medicinal chemistry, pharmacology, and medicine and familiar to those skilled in the art are used herein. Representative abbreviations and definitions are provided below.

[0032] Ac is acetyl [CH3C(O)-], Ac2O is acetic anhydride; AcOH is acetic acid; APC is 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 triethyl amine; 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 spectral detection; LiHMDS is lithium bis(trimethylsilyl) ) amide; LG is a leaving group; M is molar; mCPBA is metachloroperbenzoic acid; mmol is millimolar; 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 indicates an unspecified protecting group; Ph is phenyl; PhMe is toluene; PPh3 is triphenylphosphine; PMB is paramethoxybenzyl; rt is room temperature; RBF is 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] definition The term "abnormal" as used herein refers to being different from normal. When used to describe activity, abnormal refers to the activity being greater than or less than the average of normal control or normal non-diseased control samples. Abnormal activity may refer to an amount of activity that causes disease, and in this case, when the abnormal activity is returned to a normal amount or a non-disease-related amount (for example, by administering a compound described herein or using a method described herein), the disease or one or more disease symptoms are alleviated.

[0034] As used herein, the term "acyl" refers to the group -C(=O)-R, where R is alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, or heterocyclyl. Acyl can be optionally substituted as described herein for each individual R group.

[0035] The term "adenocarcinoma," as used herein, refers to a malignant tumor arising from glandular cells lining organs within an organism. Non-limiting examples of adenocarcinomas include non-small cell lung cancer, prostate cancer, pancreatic cancer, esophageal cancer, and colon cancer.

[0036] The term "alkanoyl," as used herein, represents a hydrogen or alkyl group attached to the parent molecular group through a carbonyl group and is exemplified by formyl (i.e., a carboxaldehyde group), acetyl, propionyl, butyryl, and isobutyryl. Unsubstituted alkanoyl groups contain 1 to 7 carbons. Alkanoyl groups can be unsubstituted or substituted as described herein for the alkyl group (e.g., optionally substituted C1-7 alkanoyl). The suffix "-(o)yl" can be attached to other groups defined herein, such as aryl, cycloalkyl, and heterocyclyl, to define "aryloyl," "cycloalkanoyl," and "(heterocyclyl)oyl." These groups represent a carbonyl group substituted with aryl, cycloalkyl, or heterocyclyl, respectively. Each of "aryloyl", "cycloalkanoyl", and "(heterocyclyl)oyl" may be optionally substituted as defined for "aryl", "cycloalkyl", or "heterocyclyl".

[0037] The term "alkenyl," as used herein, refers to a monovalent, straight- or branched-chain, 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. Alkenyl groups may be optionally substituted as defined herein for alkyl.

[0038] The term "alkenylene" as used herein refers to a divalent alkenyl group. An optionally substituted alkenylene is an alkenylene that is optionally substituted as described herein for alkenyl.

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

[0040] As used herein, the term "alkoxyalkyl" refers to a group of the formula -LOR, where L is C 1-6 alkylene and R is C 1-6An optionally substituted alkoxyalkyl is an alkoxyalkyl optionally substituted as described herein for alkyl.

[0041] The term "alkyl," as used herein, refers to a straight- or branched-chain acyclic saturated hydrocarbon group, which, if unsubstituted, has from 1 to 12 carbons, unless otherwise specified. In certain preferred embodiments, unsubstituted alkyls have from 1 to 6 carbons. Alkyl groups are exemplified by methyl; ethyl; n-propyl and isopropyl; n-butyl, sec-butyl, isobutyl, and tert-butyl; neopentyl, and the like, and may be optionally substituted, where valence allows, with one, two, three, or, in the case of alkyl groups of two or more carbons, four or more substituents independently selected from the group consisting of amino; alkoxy; aryl; aryloxy; azido; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heterocyclyl; (heterocyclyl)oxy; heteroaryl; hydroxy; nitro; thiol; silyl; cyano; alkylsulfonyl; alkylsulfinyl; alkylsulfenyl; =0; =S; -C(O)R or -SOR (where R is amino); and =NR' (where R' is H, alkyl, aryl, or heterocyclyl). Each of the substituents may itself be unsubstituted or, where valence allows, substituted with unsubstituted substituent(s) as defined herein for each respective group.

[0042] The term "alkylene" as used herein refers to a divalent alkyl group. An optionally substituted alkylene is an alkylene that is optionally substituted as described herein for alkyl.

[0043] As used herein, the term “alkylamino” refers to a group of the formula —N(R N1 )2 or -NHR N1 (In the formula, R N1is alkyl). The alkyl portion of an alkylamino may be optionally substituted as defined for alkyl. Each optional substituent of a substituted alkylamino may itself be unsubstituted or, to the extent valence allows, substituted with unsubstituted substituent(s) as defined herein for each respective group.

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

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

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

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

[0048] The term "alkynylene" as used herein refers to a divalent alkynyl group. An optionally substituted alkynylene is an alkynylene that is optionally substituted as described herein for alkynyl.

[0049] As used herein, the term "amino" refers to -N(R N1 )2, and when amino is unsubstituted, R N1are both H; if the amino is substituted, each R N1 are 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, but each R N2 is independently H, alkyl, or aryl. Each of the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) as defined herein for each respective group. In some embodiments, amino may be unsubstituted amino (i.e., —NH) or substituted amino (e.g., —NHR N1 ), in which case R N1 are independently -OH, SO2OR N2 , -SO2R N2 , -SOR N2 , -COOR N2 , optionally substituted alkyl, or optionally substituted aryl, and each R N2 may be an optionally substituted alkyl or an optionally substituted aryl. In some embodiments, the substituted amino may be an alkylamino, where the alkyl group is optionally substituted as described herein for alkyl. In some embodiments, the amino group is -NHR N1 and R N1 is an optionally substituted alkyl.

[0050] The term "aryl," as used herein, refers to a monocyclic, bicyclic, or polycyclic carbocyclic ring system having one or two aromatic rings. Aryl groups can contain 6 to 10 carbon atoms. All atoms in 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. Aryl groups can be unsubstituted or include alkyl, alkenyl, alkynyl, alkoxy, alkylsulfinyl, alkylsulfenyl, alkylsulfonyl, amino, aryl, aryloxy, azido, cycloalkyl, cycloalkoxy, cycloalkenyl, cycloalkynyl, halo, heteroalkyl, heterocyclyl, (heterocyclyl)oxy, hydroxy, nitro, thiol, silyl, or -(CH2). n -C(O)OR A and -S0R, wherein R is amino or alkyl, and R is optionally substituted with 1, 2, 3, 4, or 5 substituents independently selected from the group consisting of: -C(O)R; and -S0R, wherein R is amino or alkyl, and R is A is H or alkyl, and n is 0 or 1. Each of the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) as defined herein for each respective group.

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

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

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

[0054] The term "azido" as used herein refers to an -N3 group.

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

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

[0057] The term "carbonyl" as used herein refers to a -C(O)- group.

[0058] The term "carcinoma," as used herein, refers to a malignant new growth made up of epithelial cells tending to infiltrate surrounding tissues and give rise to metastases.

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

[0060] The terms "CCNE1" and "cyclin E1," used interchangeably herein, refer to the G1 / S phase-specific cyclin E1 (gene name CCNE1). A cell in which CCNE1 is overexpressed is one that exhibits higher CCNE1 activity compared to a cell in which CCNE1 is normally expressed. For example, a CCNE1-overexpressing cell is one that exhibits a copy number of at least three, as opposed to a diploid normal cell that has two copies. A cell that exhibits a copy number of CCNE1 greater than three is a CCNE1-overexpressing cell. 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, unless otherwise specified, 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

[0023] Cycloalkenyl groups refer to cycloalkenyl groups. Non-limiting examples of cycloalkenyl groups 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. Cycloalkenyl groups can be unsubstituted or substituted (e.g., optionally substituted cycloalkenyl), as described for cycloalkyl.

[0062] The term "cycloalkenylalkyl," as used herein, refers to an alkyl group substituted with a cycloalkenyl group, each as defined herein. The cycloalkenyl and alkyl portions may be optionally substituted as the individual groups defined herein.

[0063] The term "cycloalkenylene," as used herein, refers to a divalent cycloalkenyl group. Optionally substituted cycloalkenylene is optionally substituted as described herein for cycloalkyl.

[0064] The term "cycloalkoxy," as used herein, unless otherwise specified, refers to a chemical substituent of 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, unless otherwise specified, the term "cycloalkyl" refers to a cyclic alkyl group having 3 to 10 carbons (e.g., C 3-C10Cycloalkyl groups refer to groups having cyclic or bicyclic ring structures, such as bicyclo[pqO]alkyl, where p and q are each 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, bicyclic cycloalkyl groups can include bridged cycloalkyl structures, e.g., bicyclo[pqr]alkyl, where r is 1, 2, or 3 and p and q are each 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. Cycloalkyl groups can also be spirocyclic groups, e.g., spiro[pq]alkyl, where p and q are each 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. A cycloalkyl group can be unsubstituted or substituted with 1, 2, 3, 4, or 5 substituents (e.g., optionally substituted cycloalkenyl), which substituents are independently alkyl; alkenyl; alkynyl; alkoxy; alkylsulfinyl; alkylsulfenyl; alkylsulfonyl; amino; aryl; aryloxy; azido; cycloalkyl; cycloalkoxy; cycloalkenyl; cycloalkynyl; halo; heteroalkyl; heterocyclyl; (heterocyclyl)oxy; heteroaryl; hydroxy; nitro; thiol; silyl; cyano; ═O; ═S; —SOR (where R is optionally substituted amino); ═NR′ (where R′ is H, alkyl, aryl, or heterocyclyl); and —CON(R A )2(in the formula, each R A are independently H or alkyl, or both R Aare selected from the group consisting of: and together with the atom to which they are attached form a heterocyclyl. Each of the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) as defined herein for each respective group.

[0066] The term "cycloalkylalkyl," as used herein, refers to an alkyl group substituted with a cycloalkyl group, each as defined herein. The cycloalkyl and alkyl portions may be optionally substituted like the individual groups described herein.

[0067] The term "cycloalkylene" as used herein refers to a divalent cycloalkyl group. Optionally substituted cycloalkylene is optionally substituted as described herein for cycloalkyl.

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

[0069] "Disease" or "condition" refers to a condition or state 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. A mutant FBXW7 gene, also referred to herein as an FBXW7 gene with an inactivating mutation, fails to produce functional FBXW7 protein or produces reduced amounts of FBXW7 protein in cells.

[0071] The term "halo" as used herein refers to a halogen selected from bromine, chlorine, iodine, and fluorine.

[0072] The term "heteroalkyl," as used herein, refers to an alkyl, alkenyl, or alkynyl group that is interrupted once by one or two heteroatoms; twice, independently each time, by one or two heteroatoms; three times, independently each time, by one or two heteroatoms; or four times, independently each time, 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 can be unsubstituted or substituted (e.g., optionally substituted heteroalkyl). When a heteroalkyl is substituted and a substituent is attached to a heteroatom, the substituent is selected according to the nature and valence of the heteroatom. Thus, substituents attached to a heteroatom can include, where valence permits, ═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, wherein 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 the substituents may itself be unsubstituted or substituted with unsubstituted substituent(s) as defined herein for each group. When a heteroalkyl is substituted and the substituent is bonded to a carbon atom, the substituent is selected from those described for alkyl, unless the substituent on the carbon atom bonded to the heteroatom is Cl, Br, or I. It is understood that the carbon atom is at the terminal end of the heteroalkyl group.

[0073] The term "heteroarylalkyl," as used herein, refers to an alkyl group substituted with a heteroaryl group, each as defined herein. The heteroaryl and alkyl portions may be optionally substituted as individual groups described herein.

[0074] The term "heteroarylene" as used herein refers to a divalent heteroaryl. An optionally substituted heteroarylene is an optionally substituted heteroarylene as described herein for heteroaryl.

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

[0076] The term "heterocyclyl," as used herein, unless otherwise specified, refers to a monocyclic, bicyclic, tricyclic, or tetracyclic ring system having a fused, bridged, and / or spiro 3-, 4-, 5-, 6-, 7-, 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" refers to a monocyclic, bicyclic, tricyclic, or tetracyclic ring system having a fused or bridged 5-, 6-, 7-, or 8-membered ring containing one, two, three, or four heteroatoms independently selected from the group consisting of nitrogen, oxygen, and sulfur, unless otherwise specified. A heterocyclyl can be aromatic or non-aromatic. Non-aromatic 5-membered heterocyclyls have 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. Heterocyclyl groups, unless otherwise specified, contain 1 to 16 carbon atoms. Certain heterocyclyl groups can contain up to 9 carbon atoms. 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, etc. When a heterocyclic ring system has at least one aromatic resonance structure or at least one aromatic tautomer, such 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, etc. The term "heterocyclyl" also refers to heterocyclic compounds having bridged polycyclic structures 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 carbon rings, such as an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentane ring, a 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. Heterocyclyl groups can 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, azido, cycloalkyl, cycloalkoxy, cycloalkenyl, cycloalkynyl, halo, heteroalkyl, heterocyclyl, (heterocyclyl)oxy, hydroxy, nitro, thiol, silyl, cyano, -C(O)R or -SOR (where R is amino or alkyl), =O, =S, or =NR' (where R' is H, alkyl, aryl, or heterocyclyl). Each of the substituents can itself be unsubstituted or substituted with the unsubstituted substituent(s) defined herein for each respective group.

[0077] The term "heterocyclylalkyl," as used herein, refers to an alkyl group substituted with a heterocyclyl group, each as defined herein. The heterocyclyl and alkyl portions may be optionally substituted as individual groups described herein.

[0078] The term "heterocyclylene" as used herein refers to a divalent heterocyclyl. An optionally substituted heterocyclylene is an optionally substituted heterocyclylene as described herein for heterocyclyl.

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

[0080] The terms "hydroxyl" and "hydroxy", used interchangeably herein, refer to an --OH group.

[0081] The term "isotopically enriched" as used herein refers to a pharmaceutical active agent in which the isotopic content of one isotope at a given position in the molecule is at least 100 times greater than the natural abundance of this isotope.For example, an isotopically enriched composition for deuterium includes an active agent in which the abundance of deuterium at the position of at least one hydrogen atom is at least 100 times greater than the natural abundance of deuterium.Preferably, the isotopic enrichment of deuterium is at least 1000 times greater than the natural abundance of deuterium.More preferably, the isotopic enrichment of deuterium is at least 4000 times (for example, at least 4750 times, for example, up to 5000 times) greater than the natural abundance of deuterium.

[0082] The term "leukemia," as used herein, broadly refers to a progressive, malignant disease of the blood-forming organs, generally characterized by the incorrect proliferation and development of white blood cells and their precursors in the blood and bone marrow. Leukemias are generally classified clinically based on (1) the duration and nature of the disease (acute or chronic); (2) the cell types involved: bone marrow (myeloid), lymph (lymphoid), or monocytic; and (3) the increased or absent number of leukemic or nonleukemic (subleukemic) abnormal cells.

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

[0084] The term "melanoma" as used herein is taken to mean a tumor arising from the melanocytic 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 an inhibitor of a compound that, when exposed to the enzyme Myt1, exhibits a measured Myt1 IC 50 The term "Myt1 inhibitor" refers to a compound that reduces Myt1 activity such that the Myt1 IC is 10 μM or less (e.g., 5 μM or less or 1 μM or less). 50 can be 100 nM or less (e.g., 10 nM or less or 3 nM or less), and in some cases as little as 100 pM or 10 pM. Preferably, the Myt1 IC 50 is 1 nM to 1 μM (e.g., 1 to 750 nM, 1 to 500 nM, or 1 to 250 nM). Even more preferably, Myt1 IC 50 is less than 20 nm (for example, 1 to 20 nM).

[0087] The term "nitro" as used herein refers to the group --NO.sub.2.

[0088] The term "oxo," as used herein, refers to a divalent oxygen atom (eg, the structure of oxo may be depicted as ═O).

[0089] The term "Ph" as used herein refers to phenyl.

[0090] As used herein, the term "pharmaceutical composition" refers to a composition containing a compound described herein, formulated with a pharmaceutically acceptable excipient, and manufactured or sold with the approval of a government regulatory agency as part of a therapeutic regimen for the treatment of a disease in a mammal. Pharmaceutical compositions can be formulated, for example, for oral administration in unit dosage form (e.g., tablets, capsules, caplets, gelcaps, or syrups); for topical administration (e.g., as a cream, gel, lotion, or ointment); for intravenous administration (e.g., as a sterile solution free of particulate embolic material and in a solvent system suitable for intravenous use); or any other formulation described herein.

[0091] The terms "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier," as used interchangeably herein, refer to any ingredient (e.g., a vehicle capable of suspending or dissolving an active compound) other than the compounds described herein that has the properties of being non-toxic and non-inflammatory to a patient. Excipients can include, for example, anti-adherents, antioxidants, binders, coatings, compression aids, disintegrants, dyes (pigments), emollients, emulsifiers, bulking agents (diluents), film-forming agents or coatings, flavors, fragrances, glidants (flow enhancers), lubricants, preservatives, printing inks, adsorbents, suspending or dispersing agents, sweeteners, or water for hydration. Exemplary excipients include, but are not limited to, butylated hydroxytoluene (BHT), calcium carbonate, calcium phosphate (dibasic), calcium stearate, croscarmellose, cross-linked polyvinylpyrrolidone, citric acid, crospovidone, cysteine, ethylcellulose, gelatin, hydroxypropyl cellulose, hydroxypropylmethylcellulose, lactose, magnesium stearate, maltitol, mannitol, methionine, methylcellulose, methylparaben, microcrystalline cellulose, polyethylene glycol, polyvinylpyrrolidone, povidone, pregelatinized starch, propylparaben, retinyl palmitate, shellac, silicon dioxide, sodium carboxymethylcellulose, sodium citrate, sodium starch glycolate, sorbitol, starch (corn), stearic acid, 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 the tissues of humans and animals, within the scope of sound medical judgment, without undue toxicity, irritation, allergic response, etc., and is commensurate with 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. Salts can be prepared in situ during the final isolation and purification of the compounds described herein, or can be prepared 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-hydroxyethanesulfonate, Examples of the salts include 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, and valerate.Representative alkali and alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like, 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 a condition that is not malignant but has a tendency to become malignant.

[0094] As used herein, the term "protecting group" refers to a group intended to prevent a hydroxy, amino, or carbonyl group from participating in one or more undesired reactions during chemical synthesis. As used herein, the term "O-protecting group" refers to a group intended to prevent a hydroxy or carbonyl group from participating in one or more undesired reactions during chemical synthesis. As used herein, the term "N-protecting group" refers to a group intended to prevent a nitrogen-containing (e.g., amino, amido, NH heterocycle, or hydrazine) group from participating in one or more undesired reactions during chemical synthesis. Commonly used O-protecting and N-protecting groups are described in Greene, "Protective Groups in Organic Synthesis," 3rd Edition (John Wiley & Sons, New York, 1999), which are incorporated herein by reference. Exemplary O-protecting and N-protecting groups include alkanoyl, aryloyl, 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-isopropylsilyloxymethyl, 4,4′-dimethoxytrityl, isobutyryl, phenoxyacetyl, 4-isopropylphenoxyacetyl, dimethylformamidino, and 4-nitrobenzoyl.

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

[0096] Other O-protecting groups include 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., For example, 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] Other N-protecting groups include protected or unprotected D-amino acids, L-amino acids, or chiral auxiliaries such as D,L-amino acids, such as alanine, leucine, and phenylalanine; sulfonyl-containing groups, such as benzenesulfonyl and p-toluenesulfonyl; 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-dimethylbenzyloxycarbonyl, ... t-butyloxycarbonyl, diisopropyl-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, and the like; aryl-alkyl groups, such as benzyl, p-methoxybenzyl, 2,4-dimethoxybenzyl, triphenylmethyl, benzyloxymethyl, and the like; silyl alkyl acetal groups, such as [2-(trimethylsilyl)ethoxy]methyl; and silyl groups, such as trimethylsilyl, and the like. 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" refers to structural isomers that are readily interconvertible, often by migration of a proton. Tautomers are distinct chemical species that can be distinguished by differences in spectroscopic properties but are usually inseparable from one another. Non-limiting examples of tautomers include ketone-enol, enamine-imine, amide-imidic acid, nitroso-oxime, ketene-ynol, and amino acid-ammonium carboxylate.

[0099] In general, the term "sarcoma" refers to a tumor made up of a substance like embryonic connective tissue and usually composed of dense solid cells embedded in a fibrillar or homogeneous substance.

[0100] The term "subject," as used herein, refers to a human or non-human animal (e.g., a mammal) that has been determined by a qualified professional (e.g., a doctor or nurse) to be suffering from or at risk for a disease or condition, with or without clinical testing(s) of a sample(s) from the subject, as known in the art. Preferably, the subject is a human. Non-limiting examples of diseases and conditions include diseases with symptoms of cellular hyperproliferation, such as cancer.

[0101] As used herein, "treatment" or "treating" refers to the medical management of a subject with the intent to improve, ameliorate, stabilize, prevent, or cure a disease or condition. This term encompasses active treatment (treatment directed at ameliorating the disease or condition); causal treatment (treatment directed at the cause of the associated disease or condition); symptomatic treatment (treatment designed to alleviate the symptoms of the disease or condition); preventative treatment (treatment directed at minimizing or partially or completely suppressing the onset of the associated disease or condition); and adjunctive treatment (treatment used to supplement another therapy). [Brief explanation of the drawings]

[0102] [Figure 1A] 1 is a bar graph showing CCNE1 amplification / overexpression across sequenced tumors from the TCGA PanCancer Atlas. [Figure 1B] FIG. 1 is a scatter plot showing CCNE1 gene expression data from the TCGA PanCancer Atlas. [Figure 2A] 1 is a bar graph showing FBXW7 mutations across sequenced tumors from the TCGA PanCancer Atlas. [Figure 2B] Figure 1 shows a lollipop graph depicting the frequency of FBXW7 mutations across the gene, highlighting three common arginine hotspot mutations (R465, R479, and R505) within the third and fourth WD40 repeats that disrupt cyclin E1 substrate recognition and are classified as deleterious. [Figure 3A] 1 is a bar graph showing the results of a proliferation assay using RPE1-hTERT Cas9 TP53− / − and CCNE1 overexpressing clones treated with different dosages of Compound 133. [Figure 3B] This is a series of images showing the results of a clonogenic survival assay using RPE1-hTERT Cas9 TP53- / - and CCNE1-overexpressing clones transduced with PKMYT1 sgRNA. Infected cells were seeded at low density to measure their ability to form colonies of more than 50 cells. After 10 days of growth, colonies were stained, imaged, and quantified. Results are normalized to the survival of RPE1-hTERT Cas9 TP53- / - parental and CCNE1-overexpressing clones transduced with a nonspecific LacZ control sgRNA. [Figure 3C] 1 is a line graph showing the results of a proliferation assay using RPE1-hTERT Cas9 TP54− / − and CCNE1 overexpressing clones treated with different dosages of Compound 133. [Figure 4A]

[0033] Figure 1 is a bar graph showing the results of a clonogenic survival assay using FT282-hTERT TP53R175H and CCNE1-overexpressing cells transduced with PKMYT1 sgRNA. Infected cells were seeded at low density to measure their ability to form colonies of more than 50 cells. After 10 days of growth, colonies were stained, imaged, and quantified. Results are normalized to the viability of FT282-hTERT TP53R175H and CCNE1-overexpressing cells transduced with AAVS1 control sgRNA. [Figure 4B] FIG. 4B is a series of images showing the stained colonies described in FIG. 4A. [Figure 4C] 1 is a line graph showing the results of a proliferation assay using FT282-hTERT TP53R175H and CCNE1 overexpressing clones treated with different dosages of Compound 133. [Figure 5] Panels A, B, and C show the results of clonogenic survival assays for stable RPE1-hTERT Cas9 TP53- / - parental and CCNE1-overexpressing clones expressing either wild-type or catalytically dead FLAG-tagged PKMYT1 sgRNA-resistant ORFs. These stable cell lines were transduced with either a LacZ nonspecific sgRNA or PKMYT1 sgRNA #4, seeded at low density, and their ability to form colonies of more than 50 cells was measured. After 10 days of growth, colonies were stained, imaged, and quantified. In C, results are normalized to the survival of RPE1-hTERT Cas9 TP53- / - CCNE1-overexpressing clones transduced with a nonspecific LacZ control sgRNA and are presented as a bar graph. Both clone 2 and clone 21 behave similarly in this study. [Figure 6] Figure 1 shows the results of a proliferation assay for a panel of CCNE1 wild-type and CCNE1 amplified / overexpressing cancer cell lines treated with different dosages of compound 28. IC50 values ​​are plotted for each cell line, demonstrating that CCNE1-overexpressing cell lines show enhanced sensitivity to Myt1 inhibitors compared to CCNE1 WT cell lines. [Figure 7] Figure 1 shows the results of a proliferation assay for a panel of FBXW7 wild-type and FBXW7 mutant cancer cell lines treated with different dosages of Compound 95. IC50 values ​​are plotted for each cell line, demonstrating that FBXW7 mutant cell lines exhibit enhanced sensitivity to Myt1 inhibitors compared to the FBXW7 WT cell line. DETAILED DESCRIPTION OF THE INVENTION

[0103] In general, the present invention provides compounds, pharmaceutical compositions containing the same, methods for preparing the compounds, and methods of use. The compounds of the present invention can be Myt1 inhibitors. Such compounds can be used to inhibit Myt1 in cells, e.g., cells of a subject (e.g., cells that overexpress CCNE1 or have an inactivating mutation in the FBXW7 gene). The subject can be in need of treatment for a disease or condition, e.g., a disease or condition with symptoms of cellular hyperproliferation, e.g., cancer. The Myt1 inhibitory activity of the compounds disclosed herein is useful for treating subjects in need of cancer treatment.

[0104] Myt1 is a cell cycle-regulating kinase primarily localized in the endoplasmic reticulum and Golgi complex. It is part of the Wee family of kinases, which also includes Wee1 and Wee1b. It is involved in the negative regulation of the CDK1-cyclin B complex, which promotes cell progression from the G2 phase of the cell cycle to the mitotic (M) phase. During DNA damage, Myt1, together with Wee1 (which mediates phosphorylation of only Tyr15), promotes the phosphorylation of CDK1 (on both Tyr15 and Thr14 of CDK1) as part of the G2 checkpoint response, maintaining the kinase complex in an inactive state in G2 and preventing progression into mitosis until the damage is repaired. In addition, it has been proposed that Myt1 directly interacts with CDK1 complexes in the cytoplasm, preventing their nuclear translocation and thus inhibiting 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, ovarian, breast, and gastric cancer, with frequencies ranging from 5 to 40%. Importantly, numerous studies have confirmed that cyclin E1 is a driving factor in tumorigenesis in these conditions, and CCNE1 amplification is observed in more aggressive subtypes, including uterine carcinosarcoma (UCS, approximately 40%), uterine serous carcinoma (USC, approximately 25%), high-grade serous ovarian carcinoma (HGSOC, approximately 25%), and triple-negative breast cancer (TNBC, approximately 8%). Patients with evidence of cyclin E1 overexpression in tumor biopsies by immunohistochemistry and / or genomic copy number analysis have lower overall survival rates compared to patients with normal cyclin E1 levels. HGSOC patients with cyclin E1 overexpression have lower response rates to cisplatin, the current standard of care.

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

[0110] Cyclin E1 overexpression and / or FBXW7 loss of function are thought to promote tumorigenesis by inducing genomic instability (e.g., increased origin firing, poor nucleotide pools, transcription-replication conflict, and / or fork instability). Cyclin E1 overexpression has been shown to induce replication stress characterized by replication fork slowing or stalling and loss of heterozygosity at fragile sites. The primary mechanism by which cyclin E1 overexpression causes replication stress is increased origin firing during early S phase, followed by depletion of replication factors, including nucleotide pools. The overall reduction in replication proteins and nucleotides reduces fork progression, leading to stalling and subsequent collapse or reversal.

[0111] The compounds of the present invention include, for example, compounds of formula (I): [ka] [During the ceremony, Each of X, Y, and Z is independently N or CR 2 and; R 1 and each R 2 are independently 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, -SO2N(R 8)2, -SO2R 7A , or -QR 7B or R 1 But R 1 One R that is vicinal to 2 in combination with an optionally substituted C 3-6 forming an alkylene; R 3 and R 4 each independently represents an optionally substituted C 1-6 is alkyl or halogen; R 5 is H or -N(R 7 )2; R 6 is -C(O)NH(R 8 ), -C(O)R 7A , or -SO2R 7A and; Each R 7 are independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl 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 or -SO2R 7A or two R 7 The group, together with the atoms attached to both, forms an optionally substituted C 2-9 forming a heterocyclyl; Each R 7A are independently optionally substituted C 1-6 Alkyl, optionally substituted C 3-8 cycloalkyl, or optionally substituted C 6-10 is aryl; Each R 7B are independently hydroxyl, optionally substituted C 1-6 Alkyl, optionally substituted C 6-10 Aryl, optionally substituted C 2-9Heterocyclyl, optionally substituted C 1-9 Heteroaryl, -N(R 7 )2, -C(O)N(R 8 )2, -SO2N(R 8 )2, -SO2R 7A or optionally substituted alkoxy; Each R 8 are independently hydrogen, optionally substituted C 1-6 Alkyl, optionally substituted C 2-6 Alkoxyalkyl, optionally substituted C 6-10 Aryl C 1-6 Alkyl, optionally substituted C 6-10 Aryl, optionally substituted C 3-8 cycloalkyl or optionally substituted C 1-9 heteroaryl; or two R 8 are combined with the atoms attached to them to form an optionally substituted C 2-9 forming a heterocyclyl; Q is an 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.

[0112] Preferably, the compound of formula (I) is an atropisomer of formula (IA): [ka] where all variables are as defined herein.

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

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

[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(R 8 )2, -SO2R 7A , or -QR 7BIt can be that.

[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

[0118] The present invention includes (where possible) individual diastereomers, enantiomers, epimers, and atropisomers of the compounds disclosed herein, as well as mixtures of diastereomers and / or enantiomers thereof, including racemic mixtures. While the specific stereochemistries disclosed herein are preferred, other stereoisomers, including diastereomers, enantiomers, epimers, atropisomers, and mixtures thereof, may also have utility in treating 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 a particular molecule may exist in more than one tautomeric form, and the 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, and pharmaceutical compositions containing the compounds and a pharmaceutically acceptable carrier. Such compounds are particularly useful, for example, in certain types of cancer and for slowing 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 that include one or more other active pharmaceutical ingredients. The compounds can be used in pharmaceutical compositions in which the compounds disclosed herein, or a pharmaceutically acceptable salt thereof, are the only active ingredient.

[0122] Optical Isomers - Diastereomers - Geometric Isomers - Tautomers The compounds disclosed herein may contain, for example, 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 (e.g., enantiomers and / or diastereomers) in mixtures and as pure or partially purified compounds are intended to be included within the scope of the present invention (i.e., all possible combinations of stereocenters, either as pure compounds or in mixtures).

[0123] Some of the compounds described herein may contain bonds that are rotationally hindered, which can be advantageous in that two individual rotamers, or 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 unless specified otherwise, are meant to include both E and Z geometric isomers.

[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 Compounds of the invention can be prepared as shown in Scheme B and described herein. The chloro in intermediate B can be prepared by S N The aryl amine can be substituted with an aromatic amine under Ar conditions. Depending on the nature of the aryl amine, placement of a protecting group may be required prior to this reaction. The bromo can be substituted with malononitrile under metal-mediated conditions to generate an aminopyrrole. OBn can be hydrogenolyzed to generate key intermediate E, which can be derivatized in several different ways to obtain compounds of the invention after nitrile hydrolysis. For example, Mitsunobu or alkylation conditions can be used to obtain R 2 Alternatively, intermediate E may be converted to a triflate to give the triflate key intermediate F, which after nitrile hydrolysis can be derivatized in several different ways to give compounds of the invention. For example, metal-mediated coupling can be used to introduce a group into R 2 A group may be introduced into R 2 Depending on the nature of the group, it may be necessary to place a protecting group prior to the hydroxyl or triflate derivatization reaction. 2 If the group contains unsaturation, a hydrogenation reaction may be required to obtain the compounds of the present invention. 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 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]

[0133] Method C Compounds of the invention can be prepared as shown in Scheme C and described herein. The 2-chloro form of commercially available 3-bromo-2,6-dichloropyridine can be prepared by Scheme C using a base. NThe bromo can be displaced with malononitrile under Ar conditions. The bromo can be displaced with aromatic amines under metal-mediated conditions to generate aminoazaindoles. Depending on the nature of the aryl amine, placement of a protecting group prior to this reaction may be required. The protecting group(s) can be removed before the nitrile can be hydrolyzed to the carboxamide by treatment with acid to give intermediate G. The remaining chloro can be derivatized in several different ways to provide compounds of the invention. For example, metal-mediated coupling can be used to convert 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 chloro-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, atropisomerically pure intermediates may be isolated, which may be further derivatized to obtain the compounds of the present invention. [ka]

[0134] Method D Compounds of the invention are shown in Scheme D and can be prepared as described herein. The halogen at the 2-position of an appropriately substituted 5-nitropyridine can be 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. NIt can be substituted with an aromatic amine under Ar or palladium-mediated conditions to produce an 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 give the compounds of the present invention. If the arylamine 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, atropisomerically pure intermediates may be isolated, which may be further derivatized to give the compounds of the present invention. [ka]

[0136] Method F Compounds of the invention can be prepared as shown in Scheme F and described herein. One chloro of commercially available 2,3-dichloro-pyrazine is S N The other chloro can be substituted with malononitrile under Ar or palladium-mediated conditions. N 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 can be cleaved to provide the key intermediate H, which can be derivatized in several different ways to obtain the compounds of the invention. For example, metal-mediated coupling can be used to convert R 2 A group may be introduced into R 2 Depending on the nature of the group, it may be necessary to place a protecting group prior to the bromo-derivatization reaction. 2 If the group contains unsaturation, a hydrogenation reaction may be required to obtain the compounds of the present invention. 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. Key intermediate C can be brominated using a suitable brominating reagent, such as NBS. 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 bromo and triflate can be derivatized sequentially with different groups, or the bromo and triflate can be simultaneously derivatized with the same group. The bromo and triflate can be derivatized in several different ways to provide the compounds of the present invention. For example, metal-mediated coupling can be used to form R 1 and / or R 2 Groups may be introduced into R sequentially or simultaneously. 1 and / or R 2 Depending on the nature of the group, placement of a protecting group prior to the bromo or triflate derivatization reaction may be required. 1 and / or R 2 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]

[0139] Method I The compounds of the present invention can be prepared as shown in Scheme I and described herein. The chloro in 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 2If the group bears a protecting group, deprotection step(s) using acid, base, and / or fluorine may be required to obtain the compounds of the 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 invention. Alternatively, atropisomerically pure intermediates may be isolated, which may be further derivatized to obtain the compounds of the invention. [ka]

[0141] Method K Compounds of the invention can be prepared as shown in Scheme K and described herein. The fluoro in 3-bromo-2-fluoro-pyridine is S N The arylamine can be substituted with an aromatic amine under Ar conditions. Depending on the nature of the arylamine, a protecting group may be required prior to this reaction. The bromo can be substituted with malononitrile under palladium-mediated conditions to generate an aminoazaindole. Hydrolysis of the nitrile can be carried out under acidic or basic conditions to give the compounds of the present invention. The arylamine or R 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]

[0142] Method L Compounds of the invention can be prepared as shown in Scheme L and described herein. The triflate of key intermediate D can be derivatized in several different ways to give compounds of the invention. For example, metal-mediated coupling can be used to form R 1 A group may be introduced into R. Pyrazines can be brominated using a suitable brominating reagent such as NBS. Bromo groups can be derivatized in several different ways to give compounds of the invention. For example, metal-mediated coupling can be used to introduce R 2 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 triflate and / or bromo derivatization reaction. 1 and / or R 2 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 invention. Depending on the nature of the arylamine used to prepare intermediate D, a mixture of atropisomers may be obtained. In such cases, isolation of the atropisomer of interest may be necessary to obtain the compounds of the invention. Alternatively, an atropisomerically pure intermediate may be isolated, which may be further derivatized to obtain the compounds of the invention. [ka]

[0143] Method M Compounds of the invention can be prepared as shown in Scheme M and described herein. The nitrile of key intermediate C can be treated with a Grignard reagent to afford a ketone. 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 several different ways to afford compounds of the invention. For example, metal-mediated coupling can be used to afford 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. 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 can be prepared as shown in Scheme O and described herein. 2-Aminopyridine can be converted to 2-hydroxypyridine, which can be converted to 2-bromopyridine. 2-Bromo can be displaced with an aromatic amine under palladium-mediated conditions. Depending on the nature of the arylamine, placement of a protecting group prior to this reaction may be required. 3-Bromo can be displaced with malononitrile under palladium-mediated conditions to generate an 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, deprotection step(s) using acid, base, and / or fluoride 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, chiral chromatography may be required to isolate the atropisomer of interest 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]

[0146] Treatment method 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), such as cancers in which CCNE1 is overexpressed or which have inactivating mutations in the FBXW7 gene.

[0147] The disease or condition can have a symptom of cell hyperproliferation. For example, the disease or condition can be cancer (e.g., a cancer in which CCNE1 is overexpressed or has an inactivating mutation in the FBXW7 gene).

[0148] 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 invention can be administered alone or in admixture with a pharmaceutical carrier selected with regard to the intended route of administration and standard pharmaceutical practice. Thus, pharmaceutical compositions for use in accordance with the present invention can be formulated in a conventional manner using one or more pharmaceutically acceptable carriers, including excipients and adjuvants that facilitate processing of the compounds of the invention into formulations and that can be used in pharmaceutically acceptable carriers.

[0154] The present invention also includes pharmaceutical compositions that may contain one or more pharmaceutically acceptable carriers. In preparing the pharmaceutical compositions of the present invention, the active ingredient is typically mixed with an excipient, diluted with an excipient, or enclosed within such a carrier, for example, in the form of a capsule, sachet, paper, or other container. When the excipient functions as a diluent, it can be a solid, semi-solid, or liquid substance (e.g., normal saline) that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the composition can be in the form of a tablet, powder, lozenge, sachet, cachet, elixir, suspension, emulsion, solution, syrup, 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 composition may also contain additional agents, such as preservatives.

[0155] Excipients or carriers are selected based on the mode and route of administration.Suitable pharmaceutical carriers and pharmaceutical essentials used in pharmaceutical preparations are described in the well-known reference books in the field, Remington: The Science and Practice of Pharmacy, 21st Ed., Gennaro, Ed., Lippincott Williams & Wilkins (2005), and USP / NF (United States Pharmacopoeia and National Formulary).Examples of suitable excipients include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, and methylcellulose.Preparation may further include lubricants such as talc, magnesium stearate, and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methylbenzoate and propylhydroxybenzoate; sweeteners; 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 prepared by conventional methods, for example, by conventional mixing, dissolving, granulating, dragee-making, pulverizing, emulsifying, encapsulating, entrapping, or lyophilizing processes. Methods for preparing 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 art of pharmaceutical formulation. When preparing a formulation, the active compound can be milled to an appropriate particle size before being combined 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 by milling to, for example, about 40 mesh, allowing for a more uniform distribution in the formulation.

[0157] Dosage The dosage of the compound used in the methods described herein, or its pharmaceutically acceptable salt or prodrug, or pharmaceutical composition thereof, can vary depending on many factors, such as the pharmacodynamic properties of the compound; the method of administration; the recipient's age, health, and weight; the nature and severity of symptoms; the frequency of treatment, and, if applicable, the type of concurrent treatment; and the clearance rate of the compound in the treated animal. Those skilled in the art can determine the appropriate dosage based on the above factors. The compound used in the methods described herein is initially administered at an appropriate dosage, and the dosage can be adjusted as necessary depending on the clinical response. In general, an appropriate daily dose of the compound of the present invention is considered to be the amount of the compound that is the lowest dose effective to produce a therapeutic effect. Such an effective dose 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 multiple doses are administered, the doses can be separated from each other, for example, by 1 to 24 hours, 1 to 7 days, 1 to 4 weeks, or 1 to 12 months. The compounds can be administered according to a schedule or without a set schedule. The active compounds can be administered, for example, once, twice, three, four, five, six, seven, eight, nine, ten, eleven, or twelve times daily; every two, three, four, five, or six days; once, two, three, four, five, six, or seven times weekly; once, two, three, four, five, or six times monthly; or once, two, three, four, five, six, seven, eight, nine, ten, eleven, or 12 times yearly. It should be understood that specific dosage regimens for a particular subject will need to be adjusted over time according to the individual's needs and the professional judgment of the person administering or supervising the administration of the compositions.

[0159] Although the appropriate amount and dosing regimen will ultimately be determined by the attending physician, an effective amount of a compound of the present invention can be, for example, a total daily dose, e.g., 0.05 mg to 3000 mg, of any of the compounds described herein. Alternatively, the dose can be calculated using the patient's body weight. Such dose ranges can include, for example, 10 to 1000 mg (e.g., 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 methods of the present invention, the duration over which multiple doses of a compound of the present invention are administered to a patient can vary. For example, in some embodiments, doses of a compound of the present invention are administered to a 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 a patient over a period of, for example, 4 to 11 months or 1 to 30 years. In some embodiments, the compound is administered to a patient at the onset of symptoms. In any of these embodiments, the amount of compound administered can vary over the course of administration. If the compound is administered daily, it can be administered, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 times daily.

[0161] formulation Using any of the methods described herein, a compound identified as capable of treating any of the conditions described herein can be administered to a patient or animal in unit dosage form with a pharmaceutically acceptable diluent, carrier, or excipient. Chemical compounds for use in such therapy can be prepared and isolated by any standard techniques known to those skilled in the art of medicinal chemistry. Conventional pharmaceutical practice can be used to provide suitable formulations or compositions for administering the identified compound to a patient suffering from a disease or condition. Administration may begin before the patient exhibits symptoms.

[0162] Exemplary routes of administration of a compound (e.g., a compound of the present invention) or a pharmaceutical composition thereof 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 with a pharmaceutically acceptable carrier. Pharmaceutical formulations of the compounds described herein formulated for treating the disorders described herein are also part of the present invention.

[0163] Oral administration formulation Pharmaceutical compositions contemplated by the present invention include those formulated for oral administration ("oral dosage forms"), which may be in the form of, for example, tablets, capsules, liquid solutions or suspensions, powders, or liquid crystals or solid crystals, which contain the active ingredient(s) in admixture with non-toxic pharmaceutically acceptable excipients. Such excipients may be, for example, inert diluents or fillers (e.g., sucrose, sorbitol, sugars, mannitol, microcrystalline cellulose, starches 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, starches 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 carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethylene glycol); and lubricants, glidants, and antiadherents (e.g., magnesium stearate, zinc stearate, stearic acid, silica, hydrogenated vegetable oil, or talc). Other pharmaceutically acceptable excipients can be colorants, flavoring agents, plasticizers, humectants, buffering agents, and the like.

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

[0165] Controlled-release compositions for oral use can be constructed to release the active drug by controlling the dissolution and / or diffusion of the active drug substance. Achieving controlled release and a targeted plasma concentration versus time profile can be accomplished by any of several means. In one example, controlled release is achieved by appropriate selection of various formulation parameters and ingredients, including, for example, various types of controlled-release compositions and coatings. Examples include single- or multi-unit tablet or capsule compositions, oil solutions, suspensions, emulsions, microcapsules, microspheres, nanoparticles, patches, and liposomes. In some embodiments, the compositions include a biodegradable, pH-, and / or temperature-sensitive polymer coating.

[0166] Dissolution or diffusion controlled release can be achieved by suitable coating of tablets, capsules, pellets, or granules of the compound, or by incorporating the compound in a suitable matrix. Controlled release coatings can include one or more of the coating materials mentioned above and / or, for example, shellac, beeswax, glycowax, castor oil wax, carnauba wax, stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerol palmitostearate, ethyl cellulose, acrylic resin, dl-polylactic acid, cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methyl methacrylate, 2-hydroxymethacrylate, methacrylate hydrogel, 1,3 butylene glycol, ethylene glycol methacrylate, and / or polyethylene glycol. In controlled release matrix formulations, the matrix material may also include, for example, hydrated methylcellulose, carnauba wax and stearyl alcohol, carbopol 934, silicone, glyceryl tristearate, methyl acrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and / or halogenated fluorocarbons.

[0167] Liquid forms into which the compounds and compositions of the present invention can be incorporated for oral administration include 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] Parenteral administration formulations The compounds described herein for use in the methods of the present invention can be administered in pharmaceutically acceptable parenteral (e.g., intravenous or intramuscular) formulations described herein. 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, formulations suitable for parenteral administration include aqueous and non-aqueous sterile injection solutions, which may contain antioxidants, buffers, bacteriostats, and solutes that render the formulation isotonic with the blood of the intended recipient, as well as aqueous and non-aqueous sterile suspensions, which may include suspending agents 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 a suitable 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 may be any of five general types of preparations identified by the USP-NF as suitable for parenteral administration: (1) "drug injections": liquid preparations of, or solutions of, the drug substance (e.g., a compound of the invention); (2) "drug injections": drug substance (e.g., a compound of the invention) as a dry solid combined with a sterile vehicle suitable for parenteral administration as a drug injection; (3) "drug injection emulsions": liquid preparations of the drug substance (e.g., a compound of the invention) dissolved or dispersed in a suitable emulsion medium; (4) "drug injection suspensions": liquid preparations of the drug substance (e.g., a compound of the invention) suspended in a suitable liquid medium; and (5) "drug injection suspensions": drug substance (e.g., a compound of the invention) as a dry solid combined with a sterile vehicle suitable for parenteral administration as a drug injection suspension.

[0170] Formulations for parenteral administration include aqueous solutions of the compound prepared by suitably mixing with a surfactant, 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 ingredients 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 to a subject in combination with one or more additional agents, such as (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 thereof.

[0174] Cytotoxic agents include, for example, actinomycin-D, alemtuzumab, alitretinoin, allopurinol, altretamine, amifostine, amphotericin, amsacrine, arsenic trioxide, asparaginase, azacitidine, azathioprine, Mycobacterium bovis (BCG), bendamustine, bexarotene, bevacizumab, bleomycin, bortezomib, busulfan, capecitabine, carboplatin, carfilzomib, carmustine, cetuximab, cisplatin, chlorambucil, cladribine, clofarabine, colchicine, and crizastatin. Intaspase, cyclophosphamide, cyclosporine, cytarabine, cytochalasin B, dacarbazine, dactinomycin, darbepoetin alfa, dasatinib, daunorubicin, 1-dehydrotestosterone, denileukin, dexamethasone, dexrazoxane, dihydroxyanthracin dione, disulfiram, docetaxel, doxorubicin, emetine, epirubicin, erlotinib, epigallocatechin gallate, epoetin alfa, estramustine, ethidium bromide, etoposide, everolimus, filgrastim, finasunate, Floxuridine, fludarabine, fluorouracil (5-FU), fulvestrant, ganciclovir, geldanamycin, gemcitabine, glucocorticoids, 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, metoprine, metronidazole, mithramycin, mitomycin-C, mitoxantrone, nandrolone, nelarabine, nilotinib, nofetumomab, oprelvekin, oxaliplatin, paclitaxel, pemetrexed, pentostatin, palifermin, pamidronate, 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] The hormone therapy can be, for example, a luteinizing hormone-releasing hormone (LHRH) antagonist. The hormone therapy can be, for example, farmagon, leuproline, goserelin, buserelin, flutamide, bicalutamide, ketoconazole, aminoglutethimide, prednisone, hydroxylprogesterone caproate, medroxyprogesterone acetate, megestrol acetate, diethylstilbestrol, ethinyl estradiol, tamoxifen, testosterone propionate, fluoxymesterone, flutamide, raloxifene, droloxifene, iodoxyfene, 4-hydroxybenzoates, benzocaine, benzodiazepine ... It may be droxitamoxifen, trioxifene, keoxifene, LY117018, onapristone, toremifine citrate, megestrol acetate, exemestane, fadrozole, vorozole, letrozole, anastrozole, nilutamide, triptorelin, histrelin, abiraterone, medroxyprogesterone acetate, diethylstilbestrol, premarin, fluoxymesterone, tretinoin, fenretinide, troxacitabine, or a combination thereof.

[0183] The apoptosis-inducing agent 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 not intended to limit the present invention in any way. In the following examples, reactions were typically carried out at room temperature (rt) under a nitrogen atmosphere using anhydrous solvents (Sure / Seal™) unless otherwise noted. Reactions were followed by TLC or by injecting small aliquots into a Waters Acquity-H UPLC Class system using an Acquity UPLC HSS C18 2.1 x 30 mm column and eluting with a gradient (1.86 min) of acetonitrile (from 15% to 98%) in water (both containing 0.1% formic acid). Preparative HPLC purification was carried out on a Teledyne Isco Combi Flash® EZ Prep system using a Phenomenex Gemini® 5 μm NX-C18 110 Å 150 × 21.2 mm column (<100 mg or multiple <100 mg injections) at a flow rate of 40 mL / min over 12 minutes, or an HP C18 RediSep® Rf gold column (>100 mg), eluting with an appropriate gradient of acetonitrile-water (both containing 0.1% formic acid), unless otherwise noted. The gradient was selected based on retention times observed by reaction tracking on a Waters Acquity-H UPLC® Class system (see above). Fractions containing the desired compound were combined and finally lyophilized. Silica gel chromatography purification was carried out on a Teledyne Isco Combi Flash® Rf system using an appropriately sized RediSep® Rf silica gel column. The purity of the final compound was assessed by injecting a small aliquot onto a Waters Acquity-H UPLC Class system using an Acquity UPLC BEH C18 2.1 x 50 mm column and eluting with a gradient (7 min) of acetonitrile (from 2% to 98%) in water (both containing 0.1% formic acid).

[0188] Abbreviation Abbreviations and terms commonly used in the fields of organic chemistry, medicinal chemistry, pharmacology, and medicine and familiar to those skilled in the art are used herein. Representative abbreviations and definitions are provided 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 a compound; Conc. means 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 the time; 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 spectral detection; LiHMDS is lithium hexamethyldisilazane; M is molar concentration, mmol is millimolar; Me is methyl; MeCN is acetonitrile; MeMgBr is methylmagnesium bromide; MeMgCl is methylmagnesium chloride; MeOH is methanol; MOM is methoxymethyl; min is minutes; 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 the complex of [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) with 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 [ka] Intermediate B (5-benzyloxy-2-bromo-3-chloro-pyrazine) Step 1. To a solution of 5-bromo-6-chloro-pyrazin-2-amine (110 g, 528 mmol) in sulfuric acid (770 mL) was added sodium nitrite (40 g, 580 mmol) in several portions under mechanical stirring at 0° C. The resulting viscous mixture was stirred at 0° C. for 1 h and then slowly poured into 6 L of crushed ice containing cold water while maintaining the temperature below 30° C. The resulting precipitate was collected by filtration, washed with water, and then dried by coevaporation with toluene twice under vacuum to give 5-bromo-6-chloro-pyrazin-2-ol (104.6 g, 95% yield) as a pale yellow-brown solid.

[0190] Step 2. 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) was added benzyl bromide (48 mL, 404 mmol) dropwise. After stirring for 3 h, the suspension was filtered through Celite. The filtrate was evaporated to dryness to give a yellow oil, which was dissolved in warm EtOH. After slow addition of water under sonication, the precipitate was collected by filtration to give 5-benzyloxy-2-bromo-3-chloro-pyrazine (85.2 g, 75% yield) as a pale yellow-brown solid. 1 HNMR (400MHz, DMSO-d6) δ 8.27 (s, 1H), 7.51-7.46 (m, 2H), 7.44-7.33 (m, 3H), 5.36 (s, 2H).

[0191] [ka] Intermediate C (6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile) Step 1. To a solution of intermediate B (90 g, 300 mmol) in toluene (1350 mL) was added potassium tert-butoxide (45.0 g, 401 mmol), 3-methoxy-2,6-dimethyl-aniline (48 g, 318 mmol), Pd2(dba)3 (14.4 g, 15.7 mmol), and Xantphos (18.0 g, 31 mmol). The mixture was degassed under vacuum and backfilled with nitrogen. The resulting mixture was stirred at 80 °C for 45 min and then concentrated under vacuum. The residue was dissolved in DCM (500 mL), 200 g of silica gel was added, and the suspension was evaporated to dryness under vacuum. The residue was purified on a pad of silica gel (1 kg silica gel) eluting with a gradient of 0 to 15% EtOAc in hexane to give 5-benzyloxy-3-chloro-N-(3-methoxy-2,6-dimethyl-phenyl)pyrazin-2-amine (108.4 g, 98% yield) as a pale yellow-brown solid.

[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) in several portions. The resulting mixture was stirred for 30 minutes, after which 5-benzyloxy-3-chloro-N-(3-methoxy-2,6-dimethyl-phenyl)pyrazin-2-amine (115 g, 311 mmol) in DME (500 mL) 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 in vacuo. Water (1 L) was slowly added, and the resulting biphasic mixture was stirred with a mechanical stirrer for 18 hours. The resulting solid was filtered off, washed with water, and dried under vacuum. Trituration in DCM afforded the first batch of the desired material as a pale yellow-brown solid, which was isolated by filtration. The mother liquor was concentrated in vacuo, and the residue was purified by silica gel chromatography (dry pack) eluting with a gradient of 10-60% EtOAc in hexanes to give a second batch of desired material. 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 yellow-brown solid. 1 HNMR(400MHz,chloroform-d)δ7.60(s,1H),7.53-7.47(m,2H),7.42-7.34(m,2H),7.33-7.27(m,1H),7.21-7.1 5(m,1H),6.94(d,J=8.5Hz,1H),5.45(s,2H),4.91(s,2H),3.84(s,3H),1.90(d,J=0.7Hz,3H),1.83(s,3H). MS:[M+1]:400.4.

[0193] [ka] Intermediate D ([6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonic acid) Step 1. A solution of intermediate C (83 g, 208 mmol) in sulfuric acid (550 mL) was stirred with a mechanical stirrer for 18 hours. The viscous brown mixture was slowly poured into ice-cold water (2 L) in an ice bath while stirring with a mechanical stirrer and maintaining the internal temperature below 20°C. A pale yellow solid precipitated. The resulting suspension in the ice bath was slowly neutralized to basic pH with aqueous ammonium hydroxide (28% solution, 850 mL) while maintaining the internal temperature below 40°C. The precipitate was collected by filtration, washed with water, and dried under vacuum to give 6-amino-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (65.1 g, 96% yield) as a pale yellow-brown solid.

[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: isocratic with 75% CO and 25% MeOH; flow rate: 70 mL / min) gave intermediate D1 and intermediate D2. [ka] Intermediate D1 from chiral SFC separation of Intermediate D. Peak 1 (retention time 4.95 min, 99.77%): R-6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethylphenyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl trifluoromethanesulfonate (1.93 g) as a white fluffy solid. H NMR (400 MHz, DMSO-d) δ 7.94 (s, 1H), 7.89 (br s, 2H), 7.50 (br s, 1H), 7.28 (dt, J = 8.4, 0.8 Hz, 1H), 7.14 (d, J = 8.5 Hz, 1H), 6.80 (br s, 1H), 3.85 (s, 3H), 1.82 (d, J = 0.7 Hz, 3H), 1.74 (s, 3H). 19FNMR (376MHz, DMSO-d6) δ-72.83. MS:[M+1]:460.0. [ka] Intermediate D2 from chiral SFC separation of Intermediate D. Peak 2 (retention time 6.44 min, 99.01%): S-6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethylphenyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl trifluoromethanesulfonate (1.95 g) as a white fluffy solid. H NMR (400 MHz, DMSO-d6) δ 7.94 (s, 1H), 7.89 (br s, 2H), 7.50 (br s, 1H), 7.28 (dt, J = 8.5, 0.7 Hz, 1H), 7.14 (d, J = 8.5 Hz, 1H), 6.80 (br s, 1H), 3.85 (s, 3H), 1.82 (d, J = 0.7 Hz, 3H), 1.74 (s, 3H). 19FNMR (376MHz, DMSO-d6) δ-72.83. MS:[M+1]:460.0.

[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 HNMR(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) was added NaH (7.0 g, 175.00 mmol, 60% dispersion in mineral oil) in several portions at 0 °C. Then, 3-bromo-2,6-dichloro-pyridine (20 g, 88.15 mmol) was added, and the resulting mixture was 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 in vacuo. 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) was added Pd(dba) (1.75 g, 1.91 mmol), 5-(methoxymethoxy)-2-methyl-aniline (3.6 g, 21.53 mmol), CsCO (12.7 g), and Xantphos (1.12 g, 1.94 mmol). The mixture was degassed under vacuum and backfilled 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 (3x). 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 10–60% EtOAc in hexanes. 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 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 4 M hydrogen chloride in dioxane (4 M, 5 mL). The mixture was stirred for 30 min. The volatiles were removed in vacuo to give 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 containing mainly 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 HNMR(400MHz,DMSO-d6)δ9.77(s,1H),7.42(s,1H),7.30(d,J=8.3Hz,1H),7.16(m, 3H),6.98-6.91(m,2H),6.87(d,J=8.1Hz,1H),6.71(d,J=2.6Hz,1H),1.81(s,3H). MS:[M+1]:317.1.

[0204] [ka] Intermediate H (6-amino-3-bromo-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a suspension of 60% NaH in mineral oil (16 g, 418 mmol) in DME (600 mL) was added propanedinitrile (26.6 g, 403 mmol) dropwise with vigorous stirring. The mixture was stirred for 30 minutes, after which 2,3-dichloropyrazine (30 g, 201 mmol) was added. The reaction mixture was stirred for 3 hours and then heated to reflux for 1 hour. The DME was evaporated under vacuum, and the resulting residue was treated with cold 1 M aqueous HCl to give a yellow product, which was filtered and washed with water and a minimal amount of ethanol to give 2-(3-chloropyrazin-2-yl)propanedinitrile (34.2 g, 95% yield) as a yellow solid.

[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. 1HNMR (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) in several portions. The resulting mixture was stirred for 5 minutes, and then 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-5-nitro-pyridin-2-amine (11.6 g, 32.9 mmol) and PdCl(dppf).CHCl (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 NaSO, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0 to 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) was added EtN (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 h and then cooled to room temperature. Ethylenediamine (500 μL) was added, and the mixture was stirred for 2 h. The resulting mixture was diluted with water and extracted with DCM (2×). 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 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 on carbon (10% w / w, 400 mg, 376 μmol). The mixture was stirred under 1 atm of H for 3 h. The suspension was filtered through a pad of Celite and concentrated in vacuo to give 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 minutes, after which it was 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 using 3 x 330 silica gel columns eluting with a gradient of 0-5% EtOAc in DCM to give 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 from the 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 (6 M, 6 mL) at 80 °C. The mixture was stirred for 20 minutes, then concentrated to dryness, coevaporated with MeOH, treated with EtN, and then concentrated to dryness. The residue was purified by reverse-phase flash chromatography on a C18 cartridge eluted with CHCN / 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 of 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) was added LiOH.HO (500 mg, 11.9 mmol) and HO (27% w / w aqueous solution, 21.02 mmol, 650 μL). The mixture was stirred at 60 °C for 20 min, 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 HNMR(400MHz,DMSO-d6)δ8.21(d,J=2.0Hz,1H),7.77(d,J=2.0Hz,1H),7.20(dt,J=8.4,0.7Hz,1H) ,7.13(s,2H),7.05(d,J=8.5Hz,1H),6.83(s,2H),3.73(s,3H),1.75(d,J=0.7Hz,3H),1.65(s,3H). MS:[M+1]:469.1.

[0216] [ka] Compound 2 (6-amino-5-(3-hydroxy-2,6-dimethylphenyl)-2-(2-(pyrrolidin-2-yl)ethyl)-5H-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of intermediate D (0.25 g, 0.55 mmol) in DMF, tert-butyl 2-ethynylpyrrolidine-1-carboxylate (0.213 g, 1.08 mmol) and EtN (234 μL, 1.66 mmol) were added. Nitrogen gas was bubbled through the reaction mixture for 10 minutes. Then, CuI (10 mg, 0.054 mmol) and PdCl(PPh) (20 mg, 0.027 mmol) were added, and the reaction mixture was heated at 100° C. for 1.5 hours. The reaction mixture was cooled to room temperature, diluted with cold water, and extracted with EtOAc (3×). The combined organic layers were dried over NaSO, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with 60% EtOAc in hexanes to give 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 (0.27 g, 83% yield) as a yellow solid.

[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 HNMR(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. 1HNMR(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. 6Hz,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. 1HNMR(400MHz,DMSO-d6)δ9.76(s,1H),7.74(d,J=3.6Hz,1H),7.38-7.16(m,1H),7.06(s,3H),6.98(d,J=8.1Hz,1H),6.88(dd ,J=8.4,2.6Hz,1H),6.83(d,J=8.0Hz,1H),6.66(d,J=2.5Hz,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] [ka] Compound 18 (2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-(3-morpholinopropyl)pyrrolo[3,2-b]pyridine-3-carboxamide) Palladium on carbon (10% w / w, 6 mg) was added to a solution of 2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-(3-morpholinoprop-1-ynyl)pyrrolo[3,2-b]pyridine-3-carboxamide (20 mg, 49 μmol) in MeOH (2 mL). The mixture was stirred under a hydrogen atmosphere for 30 minutes. The resulting mixture was filtered and concentrated in vacuo to give 2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-(3-morpholinopropyl)pyrrolo[3,2-b]pyridine-3-carboxamide (17.8 mg, 88% yield) as a white solid. 1 HNMR(400MHz,DMSO-d6)δ9.74 bs,1H),8.01(d,J=4.0Hz,1H),7.24(dd,J=8.3,0.8Hz,1H),6.97(d,J=4.0Hz,1H),6.90-6.81(m,3H),6.76(d,J=8.0Hz,1H) ,6.67(d,J=8.1Hz,1H),6.63(d,J=2.5Hz,1H),3.51(m,4H),2.77-2.58(m,2H),2.28(m,6H),1.91-1.78(m,2H),1.77(s,3H). MS:[M+1]:410.2.

[0222] [ka] Compound 23 (2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-pyrimidin-2-yl-pyrrolo[3,2-b]pyridine-3-carboxamide) To a solution of intermediate G (50 mg, 0.158 mmol) in DMF (2 mL) was added tributyl(pyrimidin-2-yl)stannane (73 mg, 0.199 mmol, 60 μL), CuI (3 mg, 16 μmol), LiCl (7 mg, 165 μmol), and PdCl(dppf).CHCl (12 mg, 16 μmol). The mixture was degassed under vacuum and backfilled with nitrogen three times, then stirred at 110 °C for 18 h. The mixture was then purified by preparative HPLC to give 2-amino-1-(5-hydroxy-2-methyl-phenyl)-5-pyrimidin-2-yl-pyrrolo[3,2-b]pyridine-3-carboxamide (8 mg, 14% yield) as a yellow solid. 1 HNMR(400MHz,DMSO-d6)δ9.79(s,1H),8.90(d,J=4.8Hz,2H),8.32(d,J=3.9Hz,1H),8.02(d,J=8.3Hz,1H),7.43(t,J=4.8Hz,1H),7.28(d, J=8.3Hz,1H),7.16(d,J=3.8Hz,1H),7.05(s,2H),7.02(d,J=8.3Hz,1H),6.90(dd,J=8.4,2.5Hz,1H),6.72(d,J=2.5Hz,1H),1.82(s,3H). MS:[M+1]:361.2.

[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 similarly to compound 6) in MeOH (1 mL) was added HCl in dioxane (4 M, 0.5 mL). The mixture was stirred for 2 hours. The volatiles were removed in vacuo. 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 HNMR(400MHz,DMSO-d6)δ9.76(s,1H),9.20(s,2H),8.01-7.41(m,1H),7.26(d,J=8.4,Hz,1H),7.12(d,J=8.3H) z,1H),7.02(s,3H),6.92-6.82(m,2H),6.72-6.61(m,2H),4.11(s,2H),3.44(m2H),3.19(m,2H),1.77(s,3H). MS:[M+1]:464.2.

[0224] [ka] Compound 28 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. A mixture of tributyl(thiazol-2-yl)stannane (345 mg, 0.922 mmol, 290 μL), Intermediate D (210 mg, 0.457 mmol), CuI (11 mg, 58 μmol), LiCl (40 mg, 0.943 mmol), and PdCl(dppf).CHCl (35 mg, 45 μmol) in DMF (3 mL) was degassed under vacuum and then backfilled with nitrogen. The final mixture was stirred at 120 °C for 4 h. Volatiles were removed under vacuum. The residue was purified by silica gel chromatography eluting with a gradient of 20 to 100% EtOAc in hexanes to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide (136 mg, 75% yield) as an off-white solid.

[0225] Step 2. The same procedure used for compound 35 for OMe deprotection using BBr was carried out on the appropriate intermediate (23 mg, 58 μmol) to give a residue that 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 HNMR(400MHz,DMSO-d6)δ9.63(s,1H),8.49(s,1H),7.91(d,J=3.2Hz,1H),7.78(d,J=3.2Hz,1H),7.61(s ,2H),7.40(s,1H),7.29(s,1H),7.06(d,J=8.4Hz,1H),6.92(d,J=8.3Hz,1H),1.77(s,3H),1.69(s,3H). MS:[M+1]:381.2.

[0226] [ka] Compound 31 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-methyl-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of 6-amino-2-benzyloxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile intermediate C (4 g, 10 mmol) in DMF (40 mL) was added NBS (4 g, 10 mmol). The mixture was stirred for 1 h, diluted with water, and finally stirred for 20 min. The resulting solid was filtered, washed with water, dried under vacuum, and then purified by silica gel chromatography eluting with a gradient of 20 to 80% EtOAc in hexanes to give 6-amino-2-benzyloxy-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (3.86 g, 81% yield) as an off-white solid.

[0227] Step 2. To a solution of 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) was added Pd(PPh3)4 (80 mg, 69 μmol) and K2CO3 (800 mg, 5.79 mmol). The mixture was degassed under vacuum and backfilled with nitrogen three times. 2,4,6-Trimethyl-1,3,5,2,4,6-trioxatriborinate (712 mg, 2.84 mmol, 0.8 mL) was then added, and the final mixture was stirred at 100 °C for 18 h. The mixture was cooled to room temperature, diluted with EtOAc, 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 to 45% EtOAc in hexanes 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 HNMR(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 HNMR(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) was added PdCl(dppf).CHCl (8 mg, 10 μmol), [2-(trifluoromethyl)-4-pyridyl]boronic acid (40 mg, 0.209 mmol), and aqueous NaCO (2 M, 200 μL). The mixture was degassed under vacuum and backfilled with nitrogen. The reaction mixture was stirred at 100 °C for 4 h, 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 hexanes 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 volatiles were removed in vacuo. The residue was dissolved in MeOH and concentrated to dryness again. It was then 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. 1HNMR(400MHz,DMSO-d6)δ9.63(s,1H),8.81(m,1H),8.62(s,1H),8.48(s,1H),8.41(m,1H),7.63(s,2 H),7.45(s,1H),7.33(s,1H),7.07(d,J=8.3Hz,1H),6.93(d,J=8.3Hz,1H),1.77(s,3H),1.69(s,3H). MS:[M+1]:443.2.

[0236] [ka] 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 in a sealed vial at 130° C. for 2 hours, 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. The same procedure used for compound 35 for OMe deprotection using BBr was carried out on the appropriate intermediate (15 mg, 32 μmol) to give a residue that 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. 1HNMR(400MHz,DMSO-d6)δ9.51(s,1H),7.72(d,J=4.8Hz,1H),7.35(s,1H),7.27(s,1H),7.14-6.96(m,4H),6.87(d,J=8.2Hz,1H), 4.13(d,J=12.4Hz,2H),2.85-2.68(m,2H),2.53(d,J=4.6Hz,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] [ka] 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 3 M solution of MeMgBr in THF (6.50 mL) at 0 °C. The mixture was warmed and stirred for 18 h. An additional 3 M solution of MeMgBr in THF (4.00 mL) was added, and the mixture was stirred for an additional 5 h. 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 in vacuo. The residue was purified on a silica gel column eluted with a gradient of 0 to 30% EtOAc in hexane 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 1-[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 hours. The volatiles were removed in vacuo. 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 of 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 CsCO (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 in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0 to 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 [7-acetyl-6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (46 mg, 0.100 mmol) in DMF (1 mL) was added LiCl (9 mg, 0.212 mmol), tributyl(pyrazin-2-yl)stannane (74 mg, 0.200 mmol), and PdCl(dppf).CHCl (7 mg, 9.6 μmol). The mixture was stirred at 120 °C for 10 h. The 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 OMe deprotection using BBr, the same procedure used for compound 35 was carried out on the appropriate intermediate (38 mg, 98 μmol) to give a residue that 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 HNMR(400MHz,DMSO-d6)δ9.62(s,1H),9.55(s,1H),8.72(s,1H),8.69-8.62(m,2H),8.13(s ,2H),7.07(d,J=8.3Hz,1H),6.93(d,J=8.3Hz,1H),2.76(s,3H),1.77(s,3H),1.69(s,3H). MS:[M+1]:375.

[0243] [ka] Compound 102 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)-2-vinyl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a solution of intermediate C (800 mg, 2.00 mmol) in DMF (10 mL) was added NIS (450 mg, 2.00 mmol). The mixture was stirred for 30 min, diluted with water, and stirred for 20 min. The resulting precipitate was collected by filtration and then purified by silica gel chromatography eluting with a gradient of 20 to 60% EtOAc in hexanes to give 6-amino-2-benzyloxy-3-iodo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (820 mg, 78% yield) as an off-white solid.

[0244] Step 2. 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) was added (1,10-phenanthroline)(trifluoromethyl)copper(I) (900 mg, 2.88 mmol). The mixture was stirred at 70 °C for 4 h. The volatiles were removed in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 20 to 60% EtOAc in hexanes 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 h, 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 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 CsCO (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 afforded [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (190 mg, 61% yield) as an off-white solid.

[0247] Step 5. To a solution of [6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)-3-(trifluoromethyl)pyrrolo[2,3-b]pyrazin-2-yl] trifluoromethanesulfonate (90 mg, 0.171 mmol) in dioxane (1 mL) was added PdCl(dppf).CHCl (14 mg, 17 μmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-dioxaborolane (30 mg, 0.195 mmol), and aqueous NaCO (2 M, 100 μL). The mixture was stirred at 120 °C for 18 hours. The volatiles were removed in vacuo. 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 HNMR (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) Step 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 min, then CD3I (5.14 mmol, 320 μL) was added, and the mixture was stirred under nitrogen for 18 h to produce an off-white suspension. ZnCl2 (0.5 M in THF, 10.5 mL) was added dropwise to the mixture. After the addition, the mixture was stirred for 20 min, 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 h. 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 NaSO, 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] Step 2. A mixture of 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 HSO (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, and the residue was triturated with water and filtered. The solid was dried under vacuum 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 HNMR(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. H 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-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (5.3 mg) was obtained as an off-white solid. H 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] [ka] Compound 116 (2-amino-5-cyclopropyl-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide) Step 1. 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 (110 mg, 0.233 mmol) (described in the synthesis of Intermediate I) in water (0.5 mL) and dioxane (2 mL) was added cyclopropylboronic acid (41 mg, 0.477 mmol), CsCO (270 mg, 0.829 mmol), and PdCl(dppf).CHCl (18 mg, 22 μmol) in a sealed vial. The mixture was degassed and backfilled with nitrogen three times. The resulting mixture was heated to 100 °C and stirred for 18 h. The volatiles were removed in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0 to 60% EtOAc in hexanes to give 2-amino-5-cyclopropyl-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carbonitrile (63 mg, 81% yield) as an off-white solid.

[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 HNMR(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),1.88m,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. 1HNMR(400MHz,DMSO-d6)δ9.52(s,1H),8.12(d,J=2.2Hz,1H),8.08-7.92(m,2H),7.46(s,1H),7.00(d, J=8.2Hz,2H),6.86(d,J=8.3Hz,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. [ka] Compound 118 from chiral SFC separation of compound 116. Peak 2 (retention time 7.81 min, 98.81%): R-2-amino-5-cyclopropyl-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide (130 mg). H 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.

[0259] [ka] Compound 132 (2-amino-5-chloro-1-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide) Step 1. To a solution of 3-methoxy-2,6-dimethyl-aniline (3.61 g, 23.9 mmol) and 3-bromo-5-chloro-2-fluoro-pyridine (5.02 g, 23.9 mmol) in THF (50 mL) was added a 1 M solution of LiHMDS in THF (48 mL) dropwise over 18 min. An exotherm of 16 °C was observed. After 30 min, the reaction mixture was diluted with saturated aqueous NH4Cl and extracted with EtOAc. The organic layer was separated, washed with brine, dried over Na2SO4, filtered, and concentrated. The crude product was purified by flash chromatography (dry load) eluting with a gradient of 0 to 20% EtOAc in heptane. The fractions were combined, concentrated, and dried under vacuum to give 3-bromo-5-chloro-N-(3-methoxy-2,6-dimethyl-phenyl)pyridin-2-amine (6.58 g, 81% yield) as a pink solid.

[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 propanedinitrile (1.62 g, 24.6 mmol) in DME (15 mL). After stirring for 30 minutes, 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 PdCl(dppf).CHCl (1.08 g, 1.32 mmol) were added. The reaction mixture was flushed with nitrogen by bubbling through the solution and then 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, then co-evaporated with toluene (2x) and dried under vacuum to give 4.67 g of crude product. Purification by silica gel chromatography (dry-loaded) eluting with a gradient of 0 to 100% EtOAc in heptane. 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 ivory-colored crystalline solid.

[0261] Step 3. To a suspension of 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) was added 98% LiOH.HO (7.22 g, 172 mmol) and HO (27% w / w aqueous solution, 9.8 mL). The mixture was stirred at 60 °C for 30 min and then cooled to room temperature. Water was added dropwise (500 mL), and the solid was collected by filtration, washed with water, and air-dried. The filtrate was diluted with water to give a second crop of solid. Finally, the filtrate was extracted with EtOAc (3x). The combined organic extracts were dried over NaSO, filtered, concentrated, and then dried under vacuum to give a third crop of crude product. The combined crude material was purified by silica gel chromatography using a gradient of 50-100% EtOAc in heptane. Pure 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-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 give 2-amino-5-chloro-1-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyridine-3-carboxamide in quantitative yield.

[0262] Step 4. The same procedure used for compound 35 for OMe deprotection using BBr was used on the appropriate intermediate (3.90 g, 11.3 mmol) to give a residue that was coevaporated with MeOH (4x), dried under vacuum, triturated with saturated aqueous NaHCO3, and filtered. The crude product was purified by silica gel chromatography using a gradient of 0-20% MeOH in CHCl2 to give 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 yellow-brown solid. 1HNMR(400MHz,DMSO-d6)δ9.54(s,1H),8.14(d,J=2.2Hz,1H),7.74(d,J=2.1Hz,1H),7.14(brs ,2H),7.06(d,J=8.3Hz,1H),6.91(d,J=8.3Hz,1H),6.86(brs,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 HNMR(400MHz,DMSO-d6)δ9.54(s,1H),8.14(d,J=2.2Hz,1H),7.74(d,J=2.1Hz,1H),7.14(brs,2H),7 .06(dt,J=8.2,0.7Hz,1H),6.91(d,J=8.3Hz,1H),6.86(brs,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). 1HNMR(400MHz,DMSO-d6)δ9.54(s,1H),8.14(d,J=2.2Hz,1H),7.74(d,J=2.1Hz,1H),7.14(brs,2H),7 .06(dt,J=8.2,0.7Hz,1H),6.91(d,J=8.3Hz,1H),6.86(brs,2H),1.74(d,J=0.7Hz,3H),1.65(s,3H). MS:[M+1]:331.1.

[0264] [ka] Compound 150 (6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-phenyl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. Intermediate F (101 mg, 0.236 mmol), phenylboronic acid (29 mg, 0.24 mmol), Pd(PPh3)4 (30 mg, 0.026 mmol), and anhydrous tribasic potassium phosphate (176 mg, 0.829 mmol) were charged into a microwave vial, which was flushed with nitrogen. Dioxane (2 mL) was then added, and the vial was capped and placed in a heat block set at 90 °C. After 90 min, the reaction mixture was cooled to room temperature, diluted with water, and extracted with EtOAc (3x). 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-5-(5-methoxy-2-methyl-phenyl)-3-phenyl-pyrrolo[2,3-b]pyrazine-7-carbonitrile (30 mg, 36% yield) as an orange solid.

[0265] Step 2. For nitrile hydrolysis using sulfuric acid, the same procedure 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. 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-(5-hydroxy-2-methyl-phenyl)-3-phenyl-pyrrolo[2,3-b]pyrazine-7-carboxamide (11 mg, 57% yield) as a white fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.71(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=8.3,2.6Hz,1H),6.77(d,J=2.5Hz,1H),1.89(s,3H). MS:[M+1]:360.2.

[0267] [ka] Compound 153 (6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-(3-pyridylmethoxy)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a mixture of Intermediate E (51 mg, 0.16 mmol), 3-pyridylmethanol (41 mg, 0.38 mmol), and triphenylphosphine (62 mg, 0.24 mmol) in THF (2 mL) was added diisopropyl azodicarboxylate (47 μL, 0.24 mmol). The resulting mixture was stirred for 18 h. Additional triphenylphosphine (62 mg, 0.24 mmol), 3-pyridylmethanol (41 mg, 0.38 mmol), and diisopropyl azodicarboxylate (47 μL, 0.24 mmol) were added, and the mixture was stirred for another 2.5 h and concentrated to dryness. The crude residue was purified by silica gel chromatography eluting with a gradient of 0 to 100% EtOAc in hexanes to give 6-amino-5-[5-(methoxymethoxy)-2-methyl-phenyl]-3-(3-pyridylmethoxy)pyrrolo[2,3-b]pyrazine-7-carbonitrile (115 mg) as an impure amber gum (containing triphenylphosphine oxide), which was carried on to the next step without further purification.

[0268] Step 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 HCl in dioxane (4 M, 1.50 mL). After stirring for 75 minutes, 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 HNMR(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(brs,1H),7.04(br s,1H),7.00(brs,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. To a nitrogen-flushed microwave vial charged with Intermediate F (251 mg, 0.549 mmol), Pd(PPh3)4 (65 mg, 0.056 mmol), and CuI (43 mg, 0.023 mmol) was added 3-ethynylpyridine (72 mg, 0.698 mmol) in DMF (2.5 mL), followed by Et3N (610 μL, 4.39 mmol). The vial was capped and then transferred to a preheated heat block (120 °C). After 1 h, the reaction was concentrated in vacuo, then taken up in THF and adsorbed onto silica. The volatiles were evaporated in vacuo 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 HCl in dioxane (4 M, 3 mL). After stirring for 30 minutes, the reaction 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 sticky 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 bisHCl salt (241 mg, 0.548 mmol) was stirred in concentrated HSO (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 NHOH / HO. The solid was collected by filtration and air-dried overnight to give the crude product as an ochre 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 HNMR(400MHz,DMSO-d6)δ9.77(br s,1H),8.75(dd,J=2.3,0.9Hz,1H),8.57(dd,J=4.9,1.7Hz,1H),8.42(s,1H),7.98(dt,J=7.9,1.9Hz,1H),7.73(brs,2H),7.44(ddd,J =8.0,4.9,0.9Hz,1H),7.35(brd,J=4.4Hz,2H),7.29(d,J=8.4Hz,1H),6.94(dd,J=8.3,2.6Hz,1H),6.77(d,J=2.6Hz,1H),1.85(s,3H). MS:[M+1]:385.3.

[0273] [ka] Compound 162 (6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-[2-(3-pyridyl)ethyl]pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. 6-Amino-5-(5-hydroxy-2-methyl-phenyl)-3-[2-(3-pyridyl)ethynyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (102 mg, 0.265 mmol) and palladium on carbon (30 mg, 0.028 mmol, 10% w / w) were stirred in MeOH (3 mL) under a hydrogen atmosphere overnight, filtered through a disk filter with MeOH, concentrated, and purified by preparative HPLC to give 6-amino-5-(5-hydroxy-2-methyl-phenyl)-3-[2-(3-pyridyl)ethyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (7 mg, 7% yield) as an off-white fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ8.34(dd,J=4.8,1.7Hz,1H),8.31(dd,J=2.3,0.8Hz,1H),7.95(s,1H),7.55(ddd,J=7.8,2.4,1.7Hz,1H),7.34(brs,1H),7.30(br s,2H),7.28(dd,J=8.4,0.8Hz,1H),7.23(ddd,J=7.8,4.8,0.9Hz,1H),7.15(brs,1H),6.92(dd ,J=8.3,2.6Hz,1H),6.72(d,J=2.6Hz,1H),3.01-2.93(m,2H),2.92-2.84(m,2H),1.82(s,3H). MS:[M+1]:389.2.

[0274] [ka] Compound 164 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-dimethyl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a suspension of NaH (3.54 g, 92 mmol, 60% dispersion in mineral oil) in THF (100 mL) at 0 °C, malononitrile (3.99 g, 60.4 mmol) in THF (30 mL) was added dropwise via an addition funnel. Upon completion of the addition, the cold bath was removed and the resulting mixture was stirred at room temperature for 45 min. 2,3-Dichloro-5,6-dimethyl-pyrazine (5.49 g, 31.0 mmol) and Pd(PPh3)4 (1.76 g, 1.52 mmol) were added, and the reaction mixture was refluxed for 3.25 h, cooled to room temperature, poured into 200 mL of a 1:1 mixture of crushed ice and 1 N HCl, and extracted with DCM (3x). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated. The crude product was adsorbed onto silica using DCM / THF and purified by silica gel chromatography eluting with a gradient of 0 to 100% EtOAc in hexanes. The mixed fractions were repurified by a second silica gel chromatography using the same conditions. The clean fractions from both columns were combined, concentrated, and dried under vacuum to give 2-(3-chloro-5,6-dimethyl-pyrazin-2-yl)propanedinitrile (5.0 g, 78% yield) as an orange solid.

[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 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 min, the reaction was concentrated to dryness, resuspended in DCM, and MeOH was added (an exotherm was observed). After concentration, the crude mixture was again co-evaporated with DCM / MeOH, then carefully triturated with saturated aqueous NaHCO3 (100 mL), diluted with water, and stirred for 1.5 h. The precipitate was filtered, washed with water, air-dried, and then purified by silica gel chromatography (dry-packed) eluting with a gradient of 0–20% MeOH in DCM. Mixed fractions were combined and repurified by silica gel chromatography using the same conditions. The clean material from both columns was 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 HNMR(400MHz,DMSO-d6)δ9.57(s,1H),7.45(br s,1H),7.18-7.02(m,4H),6.93(d,J=8.3Hz,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] Chiral SFC separation of compound 164 (5.40 g, 16.6 mmol) (apparatus: Waters Prep 100 SFC-MS; column: Phenomenex Lux Cellulose-2, 30 × 250 mm, 5 μm; conditions: isocratic with 1:1 55% ACN / EtOH at 45% CO; flow rate: 70 mL / min) gave compounds 165 and 166. [ka] 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 HNMR(400MHz,DMSO-d6)δ9.57(s,1H),7.45(br s,1H),7.12(br s,1H),7.09(brs,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 HNMR(400MHz,DMSO-d6)δ9.57(s,1H),7.45(br s,1H),7.12(br s,1H),7.09(brs,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 HNMR(400MHz,DMSO-d6)δ9.58(s,1H),7.61(s,1H),7.59(brs,1H),7.32(br s,2H),7.23(brs,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 HNMR(400MHz,DMSO-d6)δ9.63(s,1H),7.92(d,J=0.6Hz,1H),7.46(brs,2H),7.37(br s,1H),7.26(brs,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 FNMR (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 fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.63(s,1H),7.92(s,1H),7.46(brs,2H),7.42-7.32(m,1H),7.30-7.19(m, 1H),7.08(d,J=8.3Hz,1H),6.94(d,J=8.2Hz,1H),1.77(s,3H),1.75(d,J=22.4Hz,6H),1.69(s,3H). 19 FNMR (376MHz, DMSO-d6) δ-137.75 (hept, J=22.1Hz). MS:[M+1]:358.2. [ka] Compound 180 from chiral SFC separation of compound 178. Peak 2 (retention time 4.00 min, 99.90%): R-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 fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.63(s,1H),7.92(s,1H),7.46(brs,2H),7.37(br s,1H),7.26(brs,1H),7.08(d,J=8.5Hz,1H),6.94(d,J=8.3Hz,1H),1.77(s,3H),1.74(d,J=22.4Hz,6H),1.69(s,3H). 19 FNMR (376MHz, DMSO-d6) δ-137.75 (hept, J=22.1Hz). MS:[M+1]:358.2.

[0283] [ka] Compound 181 (from Method D) 2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5,6-dimethyl-pyrrolo[2,3-b]pyridine-3-carboxamide Step 1. A pressure vessel was charged with 3-bromo-2-chloro-6-methyl-5-nitro-pyridine (10.11 g, 40.2 mmol) and 3-methoxy-2,6-dimethylaniline (aniline A2, 9.20 g, 60.8 mmol). NMP (40 mL) and 2,6-dimethylpyridine (8.58 g, 80.1 mmol, 9.3 mL) were added, and the reaction mixture was heated to 130 °C (pellet bath) for 5 days until satisfactory conversion was achieved, as assessed by UPLC-MS. The reaction mixture was cooled to room temperature, and the resulting paste was transferred to a conical flask. 500 mL of 0.5 N HCl was added dropwise with stirring to give a sticky viscous material. The supernatant was filtered through a Buchner funnel. The remaining viscous material was washed with HO, dissolved in DCM, and combined with the solids also dissolved in DCM (total 200 mL). The DCM solution was dried over NaSO, filtered, and concentrated. The crude residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of 0 to 100% DCM in heptane to give 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyridin-2-amine (10.5 g, 71% yield) as a pale yellow solid.

[0284] Step 2. To a RBF containing sodium hydride (3.13 g, 72.2 mmol, 60% w / w in mineral oil) in DME (150 mL) was slowly added a solution of propanedinitrile (4.75 g, 71.9 mmol) in DME (50 mL). After stirring for 1 h, 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-5-nitro-pyridin-2-amine (10.5 g, 28.7 mmol) and Pd(dppf)Cl·DCM (2.31 g, 2.83 mmol) were added. The resulting mixture was degassed by bubbling N into the solution, fitted with a condenser, and heated to 95 °C for 1 h. The reaction mixture was cooled to room temperature, poured into saturated aqueous NH4Cl, and extracted with DCM (3x). The combined organic extracts were washed with HO, brine, dried over NaSO, filtered, and adsorbed onto silica. The crude residue was purified by silica gel chromatography (dry-loaded) eluting with a gradient of 0 to 100% EtOAc in heptane. The appropriate fractions were combined and 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. A second crop of material was obtained from the filtrate from the previous trituration by similar flash chromatography and trituration to give 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. To a RBF containing 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, 25.6 mmol) (obtained from the previous step, assumed quantitative) in DCM (280 mL) and MeOH (280 mL) was added palladium on carbon (2.08 g, 1.95 mmol, 10% w / w) as a slurry made with some of the solvent mixture. The reaction mixture was flushed with H2 and stirred overnight under an H2 atmosphere (balloon). The reaction mixture was flushed with N2 and filtered through a pad of Celite, rinsing with DCM and MeOH. The filtrate was concentrated and dried under vacuum to give a pale yellow solid, which was purified by silica gel chromatography (dry pack) eluting with a gradient of 20-100% EtOAc in heptane. The appropriate fractions were combined and concentrated in vacuo to give tert-butyl N-[5-amino-3-cyano-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridin-2-yl]carbamate (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 slow addition of saturated NH4OH. The resulting solid was collected by filtration through a Buchner funnel and washed with HO. The material was air-dried, then co-evaporated twice with toluene, dried under vacuum, and 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 pink solid.

[0290] Step 8. To a solution of methylmagnesium chloride (3 M, 18.8 mL) in THF (160 mL) in a RBF under N2, zinc dichloride in THF (0.5 M, 112 mL) was added dropwise via an addition funnel at room temperature. After the 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 fitted with a condenser and refluxed (heat block set at 80 °C) for 24 hours. The reaction mixture was cooled to room temperature, then diluted with saturated aqueous NH4Cl and extracted with EtOAc (3x). The combined organic extracts were washed with brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of 0-100% EtOAc in heptane, and then purified again by silica gel chromatography (dry-packed) eluting with a gradient of 1-15% MeOH in DCM. The appropriate fractions from the two columns were combined and concentrated in vacuo to give 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 contained some 2-amino-1-(3-methoxy-2,6-dimethyl-phenyl)-6-methyl-pyrrolo[2,3-b]pyridine-3-carboxamide by-product (19% by UPLCMS).

[0291] Step 9. To a suspension of 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 tribromoborane in DCM (1 M, 26 mmol, 26 mL) dropwise. 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 (exothermic). The mixture was concentrated to dryness and then co-evaporated twice with MeOH. The residue was triturated with saturated aqueous NaHCO3. The solid was collected by filtration on a Buchner funnel, washed with HO, 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 yellow-brown solid. MS: [M+1]: 325.1. A different batch was 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 fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.51(s,1H),7.82(s,1H),7.05(d,J=8.3Hz,1H),6.90(d,J=8.2Hz,1H),6.71(brs,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: isocratic with 55% IPA + 10 mM ammonium formate at 45% CO; flow rate: 70 mL / min) gave compounds 182 and 183. [ka] Compound 182 from 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 fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.50(s,1H),7.83(s,1H),7.05(d,J=8.3Hz,1H),6.90(d,J=8. 3Hz, 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. [ka] Compound 183 from 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 fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.50(s,1H),7.83(s,1H),7.05(d,J=8.2Hz,1H),6.90(d,J=8. 2Hz, 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] [ka] 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 in one portion to obtain a solution. The solution was cooled to 0-5 °C using an ice-water bath to obtain a suspension. Under vigorous stirring, a solution of sodium nitrite (49.25 g, 713.7 mmol) in water (175.0 mL) was added dropwise over 90 min. The ice-water bath was removed, and the suspension was slowly warmed to 11 °C and stirred for 1 h. A solution of sodium hydroxide (175 g, 4.37 mol) in 400 mL of water was added dropwise to maintain the temperature below 20 °C. The pH of the solution was adjusted to 7 with KHPO (approximately 58 g, 0.33 mol) in 70 mL of water. The suspension was filtered at 10 °C. The filter cake was triturated in water (250 mL) and filtered. The filter cake was washed thoroughly with ice-cold water and dried by vacuum suction. The product was dried overnight in an oven at 60° C. under vacuum to give 3-bromo-5,6-dimethylpyridin-2-ol as a pale yellow crystalline solid (105.69 g, 91.6%). 1 HNMR (400MHz, 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 HNMR (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-neck 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). The reaction mixture was sonicated for 20 minutes while nitrogen was sparged through the suspension. 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 approximately 200 mL, and heptane (300 mL) was added. The solvent was evaporated to give a suspension with approximately 2 volumes of solvent. The suspension was filtered and washed with heptane to give 3-bromo-N-(3-methoxy-2,6-dimethylphenyl)-5,6-dimethylpyridin-2-amine as a pale yellow solid (56.2 g, 80.8%). 1 HNMR(400MHz,DMSO-d6)δ7.57(s,1H),7.26(s,1H),7.02(d,J=8.6Hz,1H),6.77(d ,J=8.3Hz,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] Step 4. To a degassed solution of malononitrile (33.3 g, 503.5 mmol) in 1,2-dimethoxyethane (1000 mL), sodium tert-butoxide (46.3 g, 482.0 mmol) was added in four portions. 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 1,1'-bis(diphenylphosphine)ferrocenepalladium(II) chloride complex in dichloromethane (14.9 g, 18.26 mmol) were added in one portion, and the suspension was heated to a 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 of 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 min and then slowly cooled to 40 °C over 75 min. The two layers were separated in a 5000 mL flask at 40 °C, 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 and the solvent was switched to EtOAc during rotavap evaporation to give 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 give 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) DME). 1 HNMR(400MHz,DMSO-d6)δ7.38(s,1H),7.22(d,J=8.4Hz,1H),7.07(d,J=8.6Hz,1H) ,6.76(brs,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 added slowly 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 to maintain 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 temperatures 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 slowly added to a solution of KHPO (100 g) and NaOH (540 g) in water (5 L) over 1.5 hours. EtOAc (1 L) was added and the biphasic mixture was stirred for 5 min to give 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 give a suspension. The suspension was filtered and rinsed 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 min, and filtered. The product was dried in an oven under reduced pressure to give 67.8 g of crude product. The compound was suspended in DMSO (350 mL, 5 volumes) and the mixture was heated to 65 °C to give a solution. The solution was slowly cooled to 28 °C by a water bath. Water (1.05 L) was added dropwise over 2 h to give a suspension. After stirring at room temperature for 5 min, the product was filtered. The solid was triturated in 100 mL of water and filtered. The filter cake was washed with 2 x 100 mL of water. The product was dried in an oven at 60 °C under vacuum to give 2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-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). 1HNMR(400MHz,DMSO-d6)δ9.47(s,1H),7.82(s,1H),7.05(d,J=8.2Hz,1H),6.90(d,J=8.2Hz, 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] [ka] Step 1. To a solution of intermediate D (1.07 g, 2.33 mmol) in DCM (9 mL) was added a solution of BBr3 in DCM (1 M, 9.3 mL, 9.3 mmol). The mixture was stirred at 0 °C for 2 h, silica was added, and the mixture was concentrated and then purified by silica gel chromatography (dry-packed) eluting with a gradient of 0 to 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] Step 3. A solution of methyl 6-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 solution of MeMgCl in THF (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), the pH was adjusted to 7-8 using 1 N HCl, and the mixture was extracted with DCM (3x). 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 20% MeOH in DCM to give 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-(1-hydroxy-1-methyl-ethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide, compound 190 (103 mg, 32% yield) as a light tan solid. 1 HNMR(400MHz,DMSO-d6)δ9.61(s,1H),7.99(s,1H),7.50(brs,1H),7.42(br s,2H),7.23(brs,1H),7.08(d,J=8.5Hz,1H),6.94(d,J=8.3Hz,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; and 2-acetyl-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide, compound 184 (31 mg, 10% yield), was obtained as a light tan solid. 1 HNMR(400MHz,DMSO-d6)δ9.83(br s,1H),8.38(d,J=1.9Hz,1H),7.68(br s,2H),7.40(s,2H),7.09(d,J=8.2Hz,1H),6.97(d,J=8.1Hz,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 HNMR(400MHz,DMSO-d6)δ9.61(br s,1H),7.99(s,1H),7.52(br d,J=3.0Hz,1H),7.32(brs,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). 1HNMR(400MHz,DMSO-d6)δ9.62(br s,1H),7.99(s,1H),7.52(br d,J=3.2Hz,1H),7.32(brs,2H),7.18(br d,J=3.1Hz,1H),7.08(dt,J=8.3,0.8Hz,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.

[0302] 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: isocratic with 40% IPA + 10 mM ammonium formate at 60% CO; flow rate: 10 mL / min) gave compounds 185 and 186. [ka] Compound 185 from chiral SFC separation of compound 184. Peak 1 (retention time 3.87 min, 100%): S-2-acetyl-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (9 mg) as a yellow fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.68(br s,1H),8.38(s,1H),7.69(br s,2H),7.41(brs,2H),7.09(dt,J=8.3,0.7Hz,1H),6.95(d,J=8.3Hz,1H),2.68(s,3H),1.77(d,J=0.8Hz,3H),1.69(s,3H). MS:[M+1]:340.2. [ka] Compound 186 from chiral SFC separation of compound 184. Peak 2 (retention time 4.21 min, 99.80%): R-2-acetyl-6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (9 mg) as a yellow fluffy solid. 1 HNMR(400MHz,DMSO-d6)δ9.69(br s,1H),8.38(s,1H),7.69(br s,2H),7.41(brs,2H),7.09(dt,J=8.2,0.7Hz,1H),6.95(d,J=8.3Hz,1H),2.68(s,3H),1.77(d,J=0.7Hz,3H),1.69(s,3H). MS:[M+1]:340.1.

[0303] [ka] Compound 198 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-[1-(trifluoromethyl)cyclopropyl]pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. A microwave vial was charged with Intermediate D (500 mg, 1.09 mmol), 4,4,6-trimethyl-2-[1-(trifluoromethyl)vinyl]-1,3,2-dioxaborinate (502 mg, 2.26 mmol), dioxane (5 mL), and aqueous NaCO (2 M, 1.63 mL, 3.26 mmol) and flushed with nitrogen (house vac, followed by nitrogen, three times). PdCl(dppf).CHCl (444 mg, 0.544 mmol) was added, the vial was flushed again, capped, transferred to a preheated (80 °C) heat block, and stirred overnight. After cooling to room temperature, the reaction mixture was filtered through Celite, washed with water and DCM, and diluted with brine. The layers were separated (phase separator). The aqueous layer was extracted with DCM (2x). The combined organic extracts were concentrated and purified by silica gel chromatography eluting with a gradient of 0 to 50% EtOAc in DCM to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-[1-(trifluoromethyl)vinyl]pyrrolo[2,3-b]pyrazine-7-carboxamide (169 mg, 38% yield) as a dark yellow gum.

[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. 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 for a total of 75 min using a reflux condenser open to air. The reaction was concentrated and then purified by silica gel chromatography eluting with a gradient of 0 to 100% EtOAc in heptane followed by a gradient of 0 to 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. 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-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 HNMR(400MHz,DMSO-d6)δ9.62(s,1H),7.88(s,1H),7.49(brs,2H),7.36(br s,1H),7.28(brs,1H),7.08(d,J=8.2Hz,1H),6.94(d,J=8.3Hz,1H),1.76(s,3H),1.69(s,3H),1.46-1.29(m,4H). 19 FNMR(376MHz,DMSO-d6)δ-66.85. MS:[M+1]:406.1.

[0307] Chiral SFC separation of compound 198 (14.5 mg, 0.358 mmol) (instrument: Mettler Toledo Minigram SFC; column: Phenomenex Lux Cellulose-2, 10 x 250 mm, 5 μm; conditions: isocratic with 1:1 40% ACN / EtOH at 60% CO2; flow rate: 10 mL / min) to give Compound 199 and Compound 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. H NMR (400 MHz, DMSO-d 6 ) δ 9.61 (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 = 8.3 Hz, 1H), 6.94 (d, J = 8.3 Hz, 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 HNMR(400MHz,DMSO-d6)δ9.61(s,1H),7.87(s,1H),7.48(brs,2H),7.36(br s,1H),7.27(brs,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. 1HNMR (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 2-[3-bromo-5-(trifluoromethyl)-2-pyridyl]propanedinitrile (50 mg, 172 μmol) in DMF (2 mL) was added Pd2dba3 (16 mg, 17 μmol), 5-(methoxymethoxy)-2-methyl-aniline (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 h, cooled to room temperature, diluted with water, and extracted with EtOAc (3x). 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 to 100% EtOAc in hexanes 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 HSO (1 mL). After 90 min, the reaction mixture was poured onto crushed ice, placed in an ice bath, and neutralized with 1:1 NHOH / HO. The precipitate was filtered, washed with water, and air-dried overnight. Purification by preparative HPLC afforded 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 HNMR(400MHz,DMSO-d6)δ9.76(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=8.5Hz,1H),6.71(s,1H),1.78(s,3H). MS:[M+1]:351.3.

[0314] [ka] Compound 214 (6-amino-3-(2-cyclopropylethynyl)-5-(3-hydroxy-2,6-dimethyl-phenyl)-2-thiazol-2-yl-pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. A solution of 6-amino-2-benzyloxy-3-bromo-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carbonitrile (400 mg, 836 μmol, as described in the preparation of compound 31) in concentrated HSO (2 mL) and DCM (2 mL) was stirred for 10 min. 0.4 M NaOH was added, followed by water, and the resulting precipitate was removed by filtration. The crude product was purified by silica gel chromatography using a gradient of 0 to 20% MeOH in DCM to give 6-amino-3-bromo-2-hydroxy-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (300 mg, 88% yield).

[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] Step 5. The same procedure as used for compound 35 for OMe deprotection using BBr3 gave a residue which 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 HNMR (400MHz, methanol-d4) δ9.16(d,J=4.2Hz,1H),8.47(d,J=4.2Hz,2H),7.89(d,J=1.1Hz,1H),7.15(d,J=8.3H) z,1H),6.98(d,J=8.3Hz,1H),2.55-2.47(m,1H),1.88(d,J=21.7Hz,6H),1.37-1.31(m,2H),1.11-1.05(m,2H). MS:[M+1]:445.3.

[0319] [ka] Compound 215 (2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carboxamide) Step 1. A microwave vial charged with 4-prop-2-ynylmorpholine (26 mg, 204 μmol), copper(I) iodide (3.5 mg, 19 μmol), and triethylamine (1.49 mmol, 207 μL) was flushed with N. Then, 6-amino-3-bromo-7-carbamoyl-5-(3-methoxy-2,6-dimethylphenyl)-5H-pyrrolo[2,3-b]pyrazin-2-yl trifluoromethanesulfonate (100 mg, 186 μmol) in DMF (1 mL) was added, followed by PdCl(PPh) (14 mg, 19 μmol). The vial was capped and heated to 120 °C. After 1 h, the mixture was concentrated in vacuo and then adsorbed onto silica (4 g) using THF. Purification by silica gel chromatography eluting with a gradient of 0-100% EtOAc in hexanes followed by a gradient of 0-20% MeOH in EtOAc gave 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-bis(3-morpholinoprop-1-ynyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (16 mg, 15% yield).

[0320] Step 2. The same procedure used for compound 35 for OMe deprotection using BBr3 gave a residue which was purified by preparative HPLC to give 2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carboxamide (7 mg, 45% yield). 1 HNMR(400MHz,DMSO-d6)δ9.63(s,1H),7.70(s,2H),7.35-7.13(m,2H),7.05(d,J=8.3Hz,1H),6.92(d, J=8.3Hz,1H),3.62-3.51(m,11H),3.49(s,2H),2.52(q,J=4.3,3.9Hz,4H),1.74(s,3H),1.66(s,3H). MS:[M+1]:544.5.

[0321] [ka] Step 1. A mixture of intermediate D (150 mg, 327 μmol), Zn(CN) (38.3 mg, 327 μmol), and Zn powder (4 mg, 65 μmol) in NMP (3 mL) was degassed. Pd(PPh) (38 mg, 33 μmol) was added, and the mixture was stirred at 120 °C for 24 h, cooled to room temperature, quenched with saturated aqueous NHCl, and extracted with EtOAc (3x). The combined extracts were washed with brine, dried over NaSO, and concentrated. The residue was purified by preparative HPLC to give 6-amino-2-cyano-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (50 mg, 46% yield).

[0322] Step 2. 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-cyano-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide, compound 219 (2.6 mg, 9% yield), which was 1 H 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; further afforded compound 220 (10 mg, 33% yield), which is 6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2,7-dicarboxamide. 1 HNMR(400MHz,DMSO-d6)δ9.57(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=8.3Hz,1H),1.73(s,3H),1.65(s,3H). MS:[M+1]:341.4.

[0323] [ka] Compound 221 (2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carboxamide) Step 1. To a solution of 3-methoxy-2,6-dimethyl-aniline (2.02 g, 13.4 mmol) in toluene (15 mL) was added potassium tert-butoxide (1.64 g, 14.6 mmol), 3-bromo-2-chloro-5-(trifluoromethyl)pyridine (3.16 g, 12.1 mmol), Pd2dba3 (582 mg, 635 μmol), and Xantphos (723 mg, 1.26 mmol). The mixture was degassed under vacuum and backfilled with nitrogen. The resulting mixture was stirred at 120 °C for 2 h. The reaction mixture was cooled to room temperature, diluted with EtOAc, washed with water, brine, dried over Na2SO4, filtered, and concentrated to dryness. The residue was purified by flash silica gel chromatography eluting with a gradient of 0 to 70% EtOAc in hexanes to give 3-bromo-N-(3-methoxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyridin-2-amine (1.55 g, 34% yield).

[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 nitrile hydrolysis using sulfuric acid, the same procedure 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. 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 2-amino-1-(3-hydroxy-2,6-dimethyl-phenyl)-5-(trifluoromethyl)pyrrolo[2,3-b]pyridine-3-carboxamide (25 mg, 59% yield). 1 HNMR(400MHz,DMSO-d6)δ9.66(s,1H),8.47(dt,J=2.0,1.0Hz,1H),7.77(s,1H),7.31( s,2H),7.23(s,1H),7.10(dt,J=8.3,0.8Hz,1H),7.00-6.80(m,2H),1.82-1.57(m,6H). MS:[M+1]:365.3.

[0327] [ka] Compound 242 (6-amino-3-[2-(4,4-difluoro-1-hydroxy-cyclohexyl)ethynyl]-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) To a microwave vial charged with intermediate H (20.0 mg, 53.2 μmol), CuI (1.0 mg, 5.3 μmol), and 1-ethynyl-4,4-difluoro-cyclohexanol (43 mg, 266 μmol) in DMF (1 mL) was added PdCl(PPh) (4 mg, 6 μmol) and EtN (425 μmol, 60 μL). The vial was capped and heated to 80 °C for 3 h. After cooling to room temperature, the mixture was filtered and purified by preparative HPLC to give 6-amino-3-[2-(4,4-difluoro-1-hydroxy-cyclohexyl)ethynyl]-5-(3-hydroxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (5.1 mg, 21% yield). 1 HNMR(400MHz,DMSO-d6)δ9.63(s,1H),8.21(s,1H),7.57(d,J=10.6Hz,2H),7.28(s,2H),7.05(d ,J=8.3Hz,1H),6.92(d,J=8.3Hz,1H),5.77(s,1H),2.11-1.74(m,8H),1.73(s,3H),1.65(s,3H). MS:[M+1]:456.4.

[0328] [ka] Compound 243 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-N2-(3-pyridyl)pyrrolo[2,3-b]pyrazine-2,7-dicarboxamide) Step 1. A mixture of intermediate D (1.00 g, 2.18 mmol) and PdCl(PPh) (319 mg, 435 μmol) in a mixture of DMF (5 mL), MeOH (5 mL), and EtN (1.90 mL, 13.6 mmol) was heated at 70 °C under a carbon monoxide atmosphere (balloon) for 18 h. The apparatus was pre-flushed with carbon monoxide once. The volatiles were evaporated in vacuo, and the residue was purified by preparative HPLC to give methyl 6-amino-7-carbamoyl-5-(3-methoxy-2,6-dimethyl-phenyl)pyrrolo[2,3-b]pyrazine-2-carboxylate (689 mg, 86% yield).

[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). 1HNMR(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 HNMR(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 HNMR(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). 1HNMR(400MHz,DMSO-d6)δ9.66(s,1H),8.29(s,1H),8.00(s,1H),7.65(s,2H),7.30 (d,J=34.1Hz,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] [ka] Compound 266 (6-amino-5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-bis(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide) Step 1. To a suspension of magnesium turnings (380 mg, 15.7 mmol) in EtO (20 mL) was added iodine (33 mg, 130 μmol). The mixture was stirred for 10 min, after which CD3I (975 μL, 15.7 mmol) was added. The mixture was stirred under a nitrogen atmosphere for 18 h to form 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 under a nitrogen atmosphere for 72 h. 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 to 80% EtOAc in hexanes to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2-(trideuteriomethyl)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-(trideuteriomethyl)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-(trideuteriomethyl)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 to it. The mixture was stirred under a nitrogen atmosphere for 18 hours to give 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-(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (496 mg, 1.22 mmol), Pd2dba3 (111 mg, 122 μmol), and tritert-butylphosphonium tetrafluoroborate (71 mg, 244 μmol) were added, and the mixture was stirred at 70 °C under a nitrogen atmosphere for 18 h. 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 to 60% EtOAc in hexanes to give 6-amino-5-(3-methoxy-2,6-dimethyl-phenyl)-2,3-bis(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (400 mg, 95% yield).

[0341] Step 4. 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-5-(3-hydroxy-2,6-dimethyl-phenyl)-2,3-bis(trideuteriomethyl)pyrrolo[2,3-b]pyrazine-7-carboxamide (65 mg, 17% yield). 1 HNMR (400MHz, DMSO-d6) δ9.55(s,1H),7.41(s,1H),7.22-6.98(m,4H),6.89(d,J=8.3Hz,1H),1.74-1.69(m,3H),1.63(s,3H). MS:[M+1]:332.2.

[0342] Arylamine Preparation Example A variety of arylamines were used to prepare the compounds of the present invention. Some of these arylamines were commercially available, and some were prepared. Some examples of such arylamines whose preparation is described herein are listed in Table 2. [Table 2]

[0343] Preparation of arylamine A1 The compounds of the present invention can be prepared from arylamine A1, which can be prepared as shown in Scheme A1 and described herein. Commercially available 4-methyl-3-nitro-phenol can be protected at O ​​by a suitable protecting group, e.g., O-MOM. The nitro can be reduced to generate arylamine A1. [ka]

[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 the 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 give 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 iron powder (44.5 g, 796 mmol). The reaction mixture was heated to 80 °C for 3.5 h, then increased 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 and diluted with EtOAc and saturated aqueous NaHCO3. The layers were separated, and the aqueous layer was back-extracted with EtOAc (2x). The combined organic extracts were washed with brine, dried over MgSO4, filtered, and concentrated to give 26.3 g of crude product as a dark brown oil, which was purified on a silica gel pad eluting with 20-30% EtOAc in hexanes. The pure fractions were combined, concentrated, and then dried under vacuum to give 5-(methoxymethoxy)-2-methyl-aniline (25.4 g, 95% yield) as a purple oil. 1 H 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 arylamine 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 (brs, 2H), 2.14 (d, J = 0.7 Hz, 3H), 2.07 (s, 3H). MS: [M+1]: 152.3.

[0350] Preparation of arylamine A3 The compounds of the present invention can be prepared from arylamine A3, which can be prepared as shown in Scheme A3 and described herein. The methoxy 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 be O-protected with a suitable protecting group, e.g., O-MOM. The nitro can be reduced to generate arylamine A3. [ka]

[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 addition funnel. The mixture was allowed to warm slowly to room temperature overnight. The reaction mixture was then slowly poured into a stirred mixture of ice, water (1 L), and KH2PO4 (75 g). The layers were separated, and the aqueous layer was extracted with DCM (2 x 500 mL). The combined organic extracts were washed with brine (500 mL), dried over MgSO4, filtered through a silica pad (375 g), eluted with DCM, concentrated, and dried under vacuum to give 2,4-dimethyl-3-nitro-phenol (18.0 g, 98% yield) as a yellow solid.

[0352] Step 2. To a suspension of 2,4-dimethyl-3-nitro-phenol (18.96 g, 113 mmol) in DCM (200 mL) was added DIPEA (23.7 mL, 136 mmol) dropwise, followed by chloro(methoxy)methane (9.5 mL, 125 mmol). 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 MgSO, filtered through silica (approximately 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 containing palladium on carbon (5.06 g, 4.76 mmol, 10% w / w) under nitrogen was added MeOH (300 mL), followed by 1-(methoxymethoxy)-2,4-dimethyl-3-nitro-benzene (20.1 g, 95.1 mmol). The flask was flushed with hydrogen and stirred under a hydrogen atmosphere for 2 days. The reaction mixture was flushed with nitrogen for 2 hours 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 H 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 arylamine 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-methoxy-phenyl)carbamate (25 g, 74.3 mmol) in dioxane (500 mL) was added trimethylboroxine (50% w / w in THF, 20.51 g, 81.7 mmol), PdCl(dppf).CHCl (5.22 g, 7.43 mmol), and aqueous NaCO (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 in vacuo. EtOAc and water were added. The organic layer was separated, washed with brine, dried over NaSO, 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-methyl-phenyl)carbamate (13.8 g, 68% yield) as a yellowish solid.

[0358] Step 4. HCl in dioxane (4 M, 100 mL) was added to a solution of tert-butyl N-(2-chloro-3-methoxy-6-methyl-phenyl)carbamate (13.8 g, 50.8 mmol) in MeOH (100 mL). After 3 h, the volatiles were evaporated to dryness in vacuo to give a white solid, to which 250 mL of EtOAc and 250 mL of saturated aqueous NaHCO3 were added under 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 in vacuo. 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-methyl-aniline (7.9 g, 91% yield) as a clear oil that solidified upon standing. 1 HNMR (400MHz, chloroform-d) δ6.95-6.79(m,1H), 6.27(dd,J=8.3,1.5Hz,1H),4.06(brs,2H),3.83(d,J=1.6Hz,3H),2.12(d,J=0.8Hz,3H). MS:[M+1]:172.2.

[0359] Preparation of arylamine A5 Compounds of the invention can be prepared from arylamine A5, which can be prepared as shown in Scheme A5 and described herein. Commercially available 3-methoxy-2-methyl-aniline can be chlorinated with a chlorinating agent to produce arylamine A5. [ka]

[0360] Step 1. To a solution of 3-methoxy-2-methyl-aniline (100 g, 729 mmol) in DCM (500 mL) at 0° C. was added NCS (98 g, 734 mmol) in four portions (each addition was made 15 min apart). 30 min after the last addition, 100 g of silica gel was added, the mixture was evaporated in vacuo, and the black residue was purified by silica gel chromatography (dry packed) eluting with a gradient of 0-10% EtOAc in hexanes 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 (brs, 2H), 3.78 (s, 3H), 2.07 (s, 3H). MS: [M+1]: 172.3.

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

[0362] To a suspension of 3-amino-2,4-dichlorophenol.HCl salt (20 g, 93.3 mmol) in DMF (150 mL) was added 1-(chloromethyl)-4-methoxy-benzene (14.0 mL, 103 mmol), tetrabutylammonium iodide (1 g, 3.00 mmol), and CsCO (64.0 g, 196 mmol). The mixture was stirred at 40 °C overnight, after which it was diluted with water, stirred for 20 min, and filtered. The precipitate was washed with water and dried under vacuum. The crude product was purified by silica gel chromatography eluting with a gradient of 0 to 100% DCM in hexane to give 2,6-dichloro-3-[(4-methoxyphenyl)methoxy]aniline (20 g, 72% yield) as an off-white solid. 1 H 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 can be prepared from key intermediates A7, A8, or A9, which are shown in Scheme A7 and can be prepared as described herein (adapted from J.AM.CHEM.SOC. 2004, 126, 1150-1160). Commercially available 3-methoxyaniline can be N-protected with a suitable protecting group, such as NH-PIV. A directed orthometalation approach can be used to obtain the appropriate R 1 A protecting group such as CH3 or CD3 can be introduced. The NH-PIV protecting group can be cleaved under acidic conditions, and the remaining ortho position to the nitrogen can be brominated with a suitable brominating reagent, such as NBS. At this point, the bromine can be displaced with a boronic ester under metal-mediated conditions, followed by the addition of a suitable R 2 , for example, with CH or CD to generate key intermediates A7, A8, or A9. Alternatively, the N of the bromoaniline may be protected with a suitable protecting group, such as NH-Boc, prior to bromo substitution. In this case, the NH-Boc can be cleaved under acidic conditions to generate key intermediates A7, A8, or A9. [ka]

[0364] Arylamine A7 Step 1. To a solution of 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) was added 2,2-dimethylpropanoyl chloride (51 mL, 416 mmol) slowly. After 1 h, 1N aqueous HCl was added and the layers were separated. The aqueous layer was back-extracted with CHCl. ​​The organic layers were combined, washed with 1N aqueous HCl, brine, then dried over NaSO, filtered, and evaporated to dryness to give 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) was added nBuLi (2.5 M, 325 mL, 813 mmol) dropwise at 0 °C. After 2 h 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 h. 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 give N-[3-methoxy-2-(trideuteriomethyl)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-(trideuteriomethyl)phenyl]carbamate (33 g, 103 mmol) in dioxane (700 mL) was added bis(pinacolato)diboron (51 g, 201 mmol), KOAc (35.5 g, 362 mmol), and PdCl(dppf).CHCl (7.6 g, 10.4 mmol). The mixture was degassed under vacuum, backfilled with nitrogen, 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 filtered through a silica gel pad (250 g) eluting with 2 L of 50% EtOAc in heptane. The filtrate was evaporated and the residue was purified by silica gel chromatography eluting with a gradient of 0 to 20% EtOAc in heptane to give tert-butyl N-[3-methoxy-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-2-(trideuteriomethyl)phenyl]carbamate (22.5 g, 59% yield), which solidified upon standing under vacuum.

[0370] Step 7. To a solution of PdCl(dppf).CHCl (5.3 g, 7.24 mmol) in DMF (500 mL) were added CDI (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-(trideuteriomethyl)phenyl]carbamate (52 g, 142 mmol), and aqueous potassium phosphate tribasic solution (2 M, 350 mL) in rapid succession. Nitrogen was bubbled through the solution for 2 minutes, and then the mixture was stirred at 80 °C under a nitrogen atmosphere for 30 minutes. It was then cooled to room temperature and EtOAc was added. The organic layer was washed with water, brine, dried over NaSO, filtered, and concentrated to dryness. The residue was purified by silica gel chromatography eluting with a gradient of 0 to 20% EtOAc in heptane to give tert-butyl N-[3-methoxy-2,6-bis(trideuteriomethyl)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 HNMR (400MHz, DMSO-d6) δ6.68(d,J=7.2Hz,1H),6.31(d,J=7.4Hz,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 of methyl 6-bromo-3-methoxy-2-nitrobenzoate (6.0 g, 20.7 mmol) in dioxane (100 mL) and aqueous NaCO (2 M, 31 mL, 62.3 mmol) was bubbled with N in a pressure vessel, followed by the addition of Pd(dppf)Cl (1.64 g, 2.01 mmol) and trimethylboroxine (6.74 g, 26.8 mmol, 7.5 mL). N was bubbled through the solution. The vessel was capped and stirred at 100 °C overnight. The reaction mixture was cooled to room temperature, poured into H O, and extracted with EtOAc (3x). The combined organic layers were washed with brine, dried over Na SO , filtered through a silica plug, and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with a gradient of 0-100% EtOAc in Hep. The appropriate fractions were combined and concentrated in vacuo to give methyl 3-methoxy-6-methyl-2-nitro-benzoate (3.06 g, 66% yield) as a pale fawn waxy solid.

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

[0379] Step 5. To a solution of methyl 2-amino-3-methoxy-6-methylbenzoate (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 copper(II) bromide (2.86 g, 12.8 mmol). The reaction mixture was stirred at 55 °C for 9 min, then cooled to room temperature, diluted with HO, and extracted with EtOAc (3x). The combined organic extracts were washed with saturated aqueous NH4Cl, brine, dried over Na2SO4, filtered, and concentrated. The residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of 0-70% EtOAc in Hep. The appropriate fractions were combined and concentrated in vacuo to give methyl 2-bromo-3-methoxy-6-methylbenzoate (1.52 g, 54% yield) as a yellow oil. 1 H 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] Step 6. To a solution of methyl 2-bromo-3-methoxy-6-methylbenzoate (1.52 g, 5.87 mmol) in MeOH (15 mL) and THF (15 mL) was added aqueous NaOH (4 M, 15 mL, 60.0 mmol) and 30% hydrogen peroxide solution (1.5 mL). The reaction mixture was stirred at 70 °C overnight and then at 90 °C for 4 days. The reaction mixture was cooled to room temperature, and the volatiles were removed in vacuo. The residue was diluted with 3 N HCl (20 mL) and extracted with CHCl3 / iPrOH (4:1, 4x). The combined organic extracts were concentrated and then dried under vacuum, and the crude product was purified by silica gel chromatography (dry-packed) eluting with a gradient of MeOH (0-10%) in CHCl2 using 1% AcOH as a modifier. The appropriate fractions were combined and concentrated in vacuo to give 2-bromo-3-methoxy-6-methyl-benzoic acid (896 mg, 62% yield) as a white solid.

[0381] Step 7. To a solution of 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), and the mixture was heated to reflux for 1 h and cooled to room temperature. The volatiles were removed in vacuo. The residue was diluted with 15% aqueous citric acid and extracted with EtOAc (3×). The combined organic layers were washed with 1 N aqueous NaOH, brine, dried over NaSO, filtered, and concentrated in vacuo. The crude product was purified by silica gel chromatography (dry packing) eluting with a gradient of 0–30% EtOAc in Hep. The appropriate fractions were combined and concentrated in vacuo 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 HCl in dioxane (4 M, 10.5 mL, 42.0 mmol). The reaction mixture was stirred at room temperature for 75 min. The solution was evaporated to dryness in vacuo, then suspended in saturated aqueous NaHCO3 and extracted with DCM (3x, 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. The appropriate fractions were combined and concentrated in vacuo 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 arylamine A11 The compounds of the present invention can be prepared from key intermediate A11, which is shown in Scheme A9 and can be prepared as described herein. Commercially available 6-bromo-3-methoxy-2-methylbenzoic acid can be converted to N-Boc under Curtius rearrangement conditions. The NH-Boc can be cleaved under acidic conditions to generate key intermediate A11. [ka]

[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 h and cooled to room temperature. The volatiles were removed in vacuo. The residue was diluted with 15% aqueous citric acid and extracted with EtOAc (3x). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (dry-packed) eluting with a gradient of 0-30% EtOAc in Hep. The appropriate fractions were combined and concentrated in vacuo 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 as is in the next step although not pure.

[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 (brs, 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 x 250 mm, 5 μm; C1B is Phenomenex Lux Cellulose-2, 30 x 250 mm, 5 μm; C2 is Chiral Technologies IA, 10 x 250 mm, 5 μm; C3 is Chiral Technologies IC, 10 x 250 mm, 5 μm; C4 is Chiral Technologies ID, 10 x 250 mm, 5 μm; C5 is Chiral Technologies IG, 10 x 250 mm, 5 μm; C6 is Chiral Technologies AS, 10 x 250 mm, 5 μm; and C7 is Phenomenex Lux Cellulose-4, 10 x 250 mm, 5 μm.

[0388] The structural assignment of the isolated atropisomers was supported by biological activity, and the biologically active enantiomer was assigned to have the (S) configuration, which was confirmed by X-ray crystallography of the major compound. [Table 3-1] [Table 3-2]

[0389] Example 2. Enzymatic assay Myt1 kinase activity was detected using a recombinant human Myt1 kinase assay, which measures ATP hydrolysis using a commercially available ADP-Glo™ Kinase Assay (ADP-Glo™ Kinase Assay, 10000 assays, #V9102, from Promega). Briefly, 5 μL of recombinant human Myt1 (insect cell-expressed full-length PKMYT1 recombinant human protein, #A33387, from Thermo Fisher, approximately 80% purity) was 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] In Table 4, the Method column indicates the preparative method described above that was used to prepare the compound.

[0391] Example 3. Genetic validation 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 to measure their ability to form colonies of fewer than 50 cells. After 10 days of growth, colonies were stained, imaged, and quantified. Using a clonogenic survival assay, we observed a profound cellular impairment 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, with the same results (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 seen in WT and CCNE1-overexpressing clones (Figure 4C). For dose-response proliferation assays using RPE1-hTERT and FT282-hTERT cell lines, cells were seeded into 96-well plates and dosed with serially diluted Myt1 inhibitors. Cells were imaged daily using an IncuCyte S3 microscope, and percent well confluency was calculated over time. The experiment was terminated when cells reached their fourth population doubling, and IC was used for the final time point. 50 Curves were plotted and the percentage confluency was calculated relative to the cell confluency in untreated wells.

[0394] A panel of 16 cancer cell lines with either normal (n=8) or elevated (n=8) levels of CCNE1 were assessed for their sensitivity to compound 28 in a cell proliferation assay (Figure 6). For 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 proliferation status of these cells was assessed using Cell Titer Glo (CTG), and IC 50 The values ​​were plotted.

[0395] Similar experiments were performed on a panel of eight cancer cell lines harboring either wild-type FBXW7 (n=5) or mutated FBXW7 (n=3), and these cells were assessed for their sensitivity to compound 95 in a cell proliferation assay (Figure 7). For 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 inhibitors. Cells were incubated once daily in an IncuCyte Images were taken using an S3 microscope and percent well confluency was calculated over time. The experiment was terminated when cells reached their fourth population doubling, and IC was used for the final time point. 50 Curves were plotted and the percentage confluency was calculated relative to the cell confluency in untreated wells.

[0396] Other embodiments Various modifications and variations 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 should be understood that the invention as claimed 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 present invention.

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

Claims

1. Compounds of formula (I): 【Chemical 1】 [During the ceremony, Each of X, Y, and Z is independently N or CR ２ and 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 -QR ７Ｂ and Each R ２ are independently 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 -QR ７Ｂ and or R １ But, R １ One R that is vicinal to ２ Combined with C ３－６ forming an alkylene; R ３ and R ４ each independently represents an optionally substituted C １－６ is alkyl or halogen; R ５ is H or -NH ２ and R ６ is -C(O)NH ２ , —C(O)CH ３ , or -SO ２ Me; Each R ７ are 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 two R ７ The group, together with the atoms to which it is attached, is an optionally substituted C ２－９ forming a heterocyclyl; Each R ７Ａ is independent, C １－６ Alkyl, C ３－８ cycloalkyl, or C ６－１０ is 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 are independently hydrogen, optionally substituted C １－６ Alkyl, optionally substituted C ６－１０ aryl or optionally substituted C １－９ is heteroaryl; Q is an 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 is an atropisomer of formula (IA): 【Chemistry 2】 2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, which is enriched for

3. X is CR ２ 3. The compound of claim 1 or claim 2, wherein:

4. The compound has the formula (II): 【Chemistry 3】 2. The compound of claim 1, having the formula:

5. The compound is an atropisomer of formula (IIA): 【Chemistry 4】 5. The compound of claim 4, or a pharmaceutically acceptable salt thereof, which is enriched for

6. The compound has the formula (III): 【Chemistry 5】 [During the ceremony, 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 10. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

7. The compound is an atropisomer of formula (IIIA): 【Chemistry 6】 7. The compound of claim 6, or a pharmaceutically acceptable salt thereof, which is enriched for

8. R ２Ａ is hydrogen, optionally substituted C １－６ alkyl, or halogen; 8. A compound according to claim 6 or claim 7, or a pharmaceutically acceptable salt thereof.

9. R ３ is optionally substituted C １－６ The compound of any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein R is alkyl.

10. R ３ The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, wherein is halogen.

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

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

13. 13. The compound of claim 10 or claim 12, wherein the halogen is chlorine.

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

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

16. R ２ 16. The compound of claim 15, or a pharmaceutically acceptable salt thereof, wherein is optionally substituted methyl or optionally substituted isopropyl.

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

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

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

20. R １ is chlorine or bromine; 20. The compound of claim 19, or a pharmaceutically acceptable salt thereof.

21. R １ is optionally substituted C １－６ 18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, which is alkyl.

22. R １ 22. The compound of claim 21, wherein is optionally substituted methyl, optionally substituted ethyl, optionally substituted isopropyl, or optionally substituted butyl, or a pharmaceutically acceptable salt thereof.

23. R １ is optionally substituted C １－９ 18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, which is heteroaryl.

24. R １ is 1,3-thiazolyl, 1,2-thiazolyl, 1,3-oxazolyl, benzo-1,3-thiazolyl, benzo-1,3-oxazolyl, indolyl, benzimidazolyl, pyridyl, imidazolyl, pyrimidyl, pyrazinyl, pyridazinyl or pyrazolyl; R １ is the optionally substituted C １－９ 24. The compound of claim 23, or a pharmaceutically acceptable salt thereof, optionally substituted with substituents defined for heteroaryl.

25. R １ is optionally substituted C ３－８ 18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, which is cycloalkyl.

26. R １ is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; R １ is the optionally substituted C ３－８ 26. The compound of claim 25, or a pharmaceutically acceptable salt thereof, optionally substituted with the substituents defined for cycloalkyl.

27. R １ is optionally substituted C ２－９ 18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, which is heterocyclyl.

28. 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; R １ is the optionally substituted C ２－９ 28. The compound of claim 27, or a pharmaceutically acceptable salt thereof, optionally substituted with substituents defined for heterocyclyl.

29. R １ 26. The compound of claim 25, or a pharmaceutically acceptable salt thereof, wherein is optionally substituted cyclohexenyl or optionally substituted cyclopentenyl.

30. R １ is optionally substituted C ６－１０ 18. The compound of any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, which is aryl.

31. R １ 31. The compound of claim 30, or a pharmaceutically acceptable salt thereof, wherein is optionally substituted phenyl.

32. R １ But, -Q-R ７Ｂ 18. The compound according to any one of claims 1 to 17, wherein:

33. Q is optionally substituted C ２－６ 33. The compound of claim 32, or a pharmaceutically acceptable salt thereof, which is alkynylene.

34. Q is optionally substituted C １－６ 33. The compound of claim 32, or a pharmaceutically acceptable salt thereof, which is alkylene.

35. Q is optionally substituted C ６－１０ 33. The compound of claim 32, or a pharmaceutically acceptable salt thereof, which is arylene.

36. R ７Ｂ is optionally substituted C ２－９ 36. The compound of any one of claims 32 to 35, or a pharmaceutically acceptable salt thereof, which is heterocyclyl.

37. R ７Ｂ is optionally substituted C ６－１０ 36. The compound of any one of claims 32 to 35, or a pharmaceutically acceptable salt thereof, which is aryl.

38. R １ but, 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 optionally substituted with one, two, or three groups independently selected from the group consisting of: —C(O)Ot-Bu; n is 0 or 1; 38. The compound according to any one of claims 1 to 37, or a pharmaceutically acceptable salt thereof.

39. R １ But -N(R ７ ) ２ 18. The compound according to any one of claims 1 to 17, wherein:

40. R １ 40. The compound of claim 39, or a pharmaceutically acceptable salt thereof, wherein is diethylamino.

41. R ５ 41. The compound of any one of claims 1 to 40, or a pharmaceutically acceptable salt thereof, wherein is hydrogen.

42. R ５ But -NH ２ 42. The compound according to any one of claims 1 to 41, wherein:

43. R ６ is -C(O)NH ２ 43. The compound according to any one of claims 1 to 42, wherein:

44. R ６ But, -SO ２ 44. The compound of any one of claims 1 to 43, or a pharmaceutically acceptable salt thereof, wherein:

45. A compound selected from the group consisting of the following compounds, 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】

46. The compound is 【Chemistry 7】 46. ​​The compound of claim 45, wherein:

47. A compound according to any one of claims 1 to 46 or a pharmaceutically acceptable salt thereof, and pharmaceutically acceptable excipients A pharmaceutical composition comprising:

48. 48. The pharmaceutical composition of claim 47, wherein the compound is isotopically enriched with deuterium.

49. The compound 【Chemistry 8】 or a pharmaceutically acceptable salt thereof.

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