Tricyclic compounds as inhibitors of KRAS

Abstract

Disclosed are compounds of formula I, methods of using the compounds to inhibit KRAS activity, and pharmaceutical compositions containing such compounds, which are useful for treating, preventing, or ameliorating diseases or disorders associated with KRAS activity, such as cancer.

Description

[Technical field]

[0001] Related Applications This application is related to U.S. Provisional Application No. 63 / 219,274, filed July 7, 2021, U.S. Provisional Application No. 63 / 292,774, filed December 22, 2021, and U.S. Provisional Application No. 63 / 310,811, filed February 16, 2022, the contents of which are incorporated in their entirety.

[0002] The present disclosure provides compounds, as well as compositions and methods of use thereof, that modulate KRAS activity and are useful in the treatment of a variety of diseases, including cancer. [Background technology]

[0003] Ras proteins are part of a family of small GTPases that are activated by growth factors and various extracellular stimuli. The Ras family regulates intracellular signaling pathways that are involved in cell growth, migration, survival, and differentiation. Activation of RAS proteins in the cell membrane leads to the binding of key effectors and the initiation of a cascade of intracellular signaling pathways within the cell, including the RAF and PI3K kinase pathways. Somatic mutations in RAS can lead to uncontrolled cell growth and malignant transformation, while activation of RAS proteins is tightly regulated in normal cells (Simanshu, D. et al. Cell 170.1 (2017): 17-33).

[0004] The Ras family consists of three members: KRAS, NRAS, and HRAS. RAS-mutated cancers account for approximately 25% of human cancers. KRAS is the most frequently mutated isoform, accounting for 85% of all RAS mutations, while NRAS and HRAS are mutated in 12% and 3% of all RAS-mutated cancers, respectively (Simanshu, D. et al. Cell 170.1 (2017): 17-33). KRAS mutations are prevalent among the top three most lethal types of cancer: pancreatic (97%), colorectal (44%), and lung (30%) (Cox, AD et al. Nat Rev Drug Discov (2014) 13: 828-51). The majority of RAS mutations occur at amino acid residues 12, 13, and 61. The frequency of certain mutations varies between RAS gene isoforms, with G12 and Q61 mutations predominating in KRAS and NRAS, respectively, and G12, G13 and Q61 mutations occurring most frequently in HRAS. Furthermore, the spectrum of mutations in RAS isoforms differs between cancer types. For example, KRAS G12D mutations predominate in pancreatic cancer (51%), followed by colorectal adenocarcinoma (45%) and lung cancer (17%), while KRAS G12V mutations are associated with pancreatic cancer (30%), followed by colorectal adenocarcinoma (27%) and lung adenocarcinoma (23%) (Cox, AD et al. Nat Rev Drug Discov (2014) 13:828-51). In contrast, KRAS G12C mutations predominate in non-small cell lung cancer (NSCLC), accounting for 11-16% of lung adenocarcinomas and 2-5% of pancreatic and colorectal adenocarcinomas (Cox, AD et al. Nat. Rev. Drug Discov. (2014) 13:828-51). Genomic studies across hundreds of cancer cell lines have demonstrated that cancer cells containing KRAS mutations are highly dependent on KRAS function for cell growth and survival (McDonald, R. et al. Cell 170 (2017):577-592).The role of mutant KRAS as an oncogenic driver is further supported by extensive in vivo experimental evidence showing that mutant KRAS is required for the initiation and maintenance of initial tumors in animal models (Cox, AD et al. Nat Rev Drug Discov (2014) 13:828-51).

[0005] Taken together, these findings suggest that KRAS mutations play an important role in human cancer; therefore, the development of inhibitors targeting mutant KRAS may be useful in the clinical treatment of diseases characterized by KRAS mutations. Summary of the Invention

[0006] The present disclosure relates, inter alia, to a compound of formula I: [ka] or a pharma- ceutically acceptable salt thereof, wherein the constituent variables are as defined herein.

[0007] The present disclosure further provides a pharmaceutical composition comprising a compound of the present disclosure, or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient.

[0008] The present disclosure further provides a method for inhibiting KRAS activity, comprising administering to an individual a compound of the present disclosure or a pharma- ceutical acceptable salt thereof.The present disclosure also provides the use of the compound described herein in the manufacture of a medicament for use in a therapeutic method.The present disclosure also provides the compound described herein for use in a therapeutic method.

[0009] The present disclosure further provides a method of treating a disease or disorder in a patient, comprising administering to the patient a therapeutically effective amount of a compound of the present disclosure, or a pharma- ceutically acceptable salt thereof. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] compound In one embodiment, the compound of formula (I): [ka] Provided herein is a compound having the formula: Y is N or CH; R 1 is selected from Cl, CH3, CH2F, CHF2, and CF3; Cy 1 teeth, [ka] Selected from R 2 is selected from F and Cl, R 3 teeth, [ka] Selected from Cy 2 teeth, [ka] Selected from With the proviso that the compound of formula I is 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile It is unexpected.

[0011] In one embodiment of Formula I, or a pharma- ceutically acceptable salt thereof, Y is N or CH; R 1 is selected from Cl, CH3, CH2F, CHF2, and CF3; Cy 1 teeth, [ka] Selected from R 2 is selected from F and Cl, R 3 teeth, [ka] Selected from Cy 2 teeth, [ka] Selected from With the proviso that the compound of formula I is 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile It is unexpected.

[0012] In yet another embodiment, the compound of formula I has formula II: [ka] or a pharma- ceutically acceptable salt thereof, wherein R 1 is selected from Cl and CH3, Cy 1 teeth, [ka] Selected from R 3 teeth, [ka] Selected from Cy 2 teeth, [ka] is selected from.

[0013] In one embodiment of Formula I, or a pharma- ceutically acceptable salt thereof, Y is N or CH; R 1 is selected from Cl, CH3, CH2F, CHF2, and CF3; Cy 1 teeth, [ka] Selected from R 2 is selected from F and Cl, R3 teeth, [ka] Selected from Cy 2 teeth, [ka] is selected from.

[0014] In one embodiment, Y is CH. In one embodiment, Y is N.

[0015] In one embodiment, Cy 1 Cy 1 -c, Cy 1 -l, Cy 1 -m, Cy 1 -n, Cy 1 -o, Cy 1 -p, Cy 1 -q, Cy 1 -r, Cy 1 -s, and Cy 1 In one embodiment, Cy 1 Cy 1 -l, Cy 1 -m, Cy 1 -n, Cy 1 -o, Cy 1 -p, Cy 1 -q, Cy 1 -r, Cy 1 -s, and Cy 1 In one embodiment, Cy 1 Cy 1 -c, Cy 1 -m, Cy 1 -n, Cy 1 -o, Cy 1 -p, Cy 1 -q, Cy 1 -r, Cy 1 -s, and Cy 1 In one embodiment, Cy 1 Cy 1 -f, Cy 1 -g, Cy 1 -h, Cy1 -i, Cy 1 -j, Cy 1 -k and Cy 1 In one embodiment, Cy 1 Cy 1 -a, Cy 1 -m, Cy 1 -n, Cy 1 -o, Cy 1 -p, and Cy 1 In one embodiment, Cy is selected from 1 Cy 1 -c, Cy 1 -d, Cy 1 -e, Cy 1 -r, Cy 1 -s, and Cy 1 -t is selected.

[0016] In one embodiment, Cy 1 Cy 1 -a, Cy 1 -c and Cy 1 In one embodiment, Cy is selected from 1 Cy 1 -c and Cy 1 In one embodiment, Cy 1 Cy 1 -s and Cy 1 -t is selected.

[0017] In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy1 Cy 1 In one embodiment, Cy 1 Cy 1 -t.

[0018] In one embodiment, Cy 1 Cy 1 -a, Cy 1 -b, Cy 1 -c, Cy 1 -d, Cy 1 -e, Cy 1 -f, Cy 1 -g, Cy 1 -i, and Cy 1 In one embodiment, Cy 1 Cy 1 -a, Cy 1 -b, Cy 1 -c, Cy 1 -d, and Cy 1 In one embodiment, Cy is selected from 1 Cy 1 -f, Cy 1 -g, Cy 1 -h, Cy 1 -i, Cy 1 -j, and Cy 1 In one embodiment, Cy 1 Cy 1 -c, Cy 1 -d, and Cy 1 In one embodiment, Cy is selected from 1 Cy 1 -a, Cy 1 -b, and Cy 1 In one embodiment, Cy 1 Cy 1 -a and Cy 1 In one embodiment, Cy 1 Cy 1 -h and Cy 1 -k is selected.

[0019] In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 In one embodiment, Cy 1 Cy 1 -k.

[0020] In one embodiment, R 1 is selected from CH3, CH2F, CHF2, and CF3. 1 is selected from Cl, CH3, CHF2, and CF3. In one embodiment, R 1 is selected from CH2F, CHF2, and CF3. In one embodiment, R 1 is selected from Cl, CH2F, CHF2, and CF3. In one embodiment, R 1 is selected from Cl and CH3. In one embodiment, R 1 is selected from Cl and CF. In one embodiment, R 1 is selected from CH3 and CF3. In one embodiment, R 1 is Cl. In one embodiment, R 1 is CHF. In one embodiment, R 1 is CHF2. In one embodiment, R 1 is CH3. In one embodiment, R 1 is CF3.

[0021] In one embodiment, R2 is F. In one embodiment, R 2 is Cl.

[0022] In one embodiment, R 3 is R 3 -a and R 3 In one embodiment, R 3 is R 3 -b and R 3 In one embodiment, R 3 is R 3 -a and R 3 In one embodiment, R 3 is R 3 In one embodiment, R 3 is R 3 In one embodiment, R 3 is R 3 -c.

[0023] In one embodiment, R 3 is R 3 -b, R 3 -c, and R 3 In one embodiment, R 3 is R 3 -b and R 3 In one embodiment, R 3 is R 3 -c and R 3 In one embodiment, R 3 is R 3 -a and R 3 In one embodiment, R 3 is R 3 -d.

[0024] In one embodiment, Cy 2 Cy 2 -b, Cy 2 -d, Cy 2 -e, and Cy 2 In one embodiment, Cy is selected from 2 Cy 2 -b and Cy 2In one embodiment, Cy 2 Cy 2 -c and Cy 2 -d is selected.

[0025] In one embodiment, Cy 2 Cy 2 -a, Cy 2 -c and Cy 2 In one embodiment, Cy 2 Cy 2 -a, Cy 2 -b, and Cy 2 In one embodiment, Cy 2 Cy 2 -a, Cy 2 -b, and Cy 2 In one embodiment, Cy 2 Cy 2 -d, Cy 2 -e, and Cy 2 In one embodiment, Cy is selected from 2 Cy 2 -a and Cy 2 In one embodiment, Cy 2 Cy 2 -c and Cy 2 In one embodiment, Cy is selected from 2 Cy 2 -e and Cy 2 Selected from -f.

[0026] In one embodiment, Cy 2 Cy 2 In one embodiment, Cy 2 Cy 2 In one embodiment, Cy 2 Cy 2 In one embodiment, Cy 2 Cy 2 In one embodiment, Cy 2 Cy 2 In one embodiment, Cy 2 Cy 2 -f.

[0027] In another embodiment, the compound of formula I is 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile It is unexpected.

[0028] In one embodiment, the compound of formula I is 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile It is unexpected.

[0029] In another embodiment, the compound of the formulas herein is a compound of such formula, or a pharma- ceutically acceptable salt thereof.

[0030] In one embodiment, the compound of formula I is 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-1-yl)prop-2-en-1-one, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 8-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 8-(6-fluoro-1-(1-((E)-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 8-(1-((2S,4S)-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, or a pharma- ceutical acceptable salt thereof is selected from.

[0031] In another embodiment, the compound of formula I is 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-1-yl)prop-2-en-1-one, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 8-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 8-(6-fluoro-1-(1-((E)-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 8-(1-((2S,4S)-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, or a pharma- ceutical acceptable salt thereof is selected from.

[0032] In another embodiment, the compound of formula I is 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indazol-3-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile, 2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile, 2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(8-chloro-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, or a pharma- ceutical acceptable salt thereof is selected from.

[0033] In one embodiment, the compound of formula I is 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, or a pharma- ceutical acceptable salt thereof is selected from.

[0034] It is further recognized that certain features of the invention that are described for clarity in the context of separate embodiments may also be provided in combination in a single embodiment (wherein the embodiments are intended to be combined as if described in multiple sub-formulas). Conversely, various features of the invention that are described for brevity in the context of a single embodiment may also be provided individually or in any suitable subcombination. Thus, it is contemplated that the features described as embodiments of compounds of formula I may be combined in any suitable combination.

[0035] The compounds described herein may be asymmetric (e.g., have one or more stereocenters). All stereoisomers, e.g., enantiomers and diastereomers, are intended unless otherwise indicated. Compounds of the present invention that contain asymmetrically substituted carbon atoms can be isolated in optically active or racemic forms. Methods for preparing optically active forms from optically inactive starting materials are known in the art, for example, by resolution of racemic mixtures or by stereoselective synthesis. Many geometric isomers of olefins, such as C=N double bonds, may also exist in the compounds described herein, and all such stable isomers are contemplated in the present invention. Cis and trans geometric isomers of the compounds of the present invention are described and can be isolated as a mixture of isomers or as separated isomeric forms.

[0036] The resolution of a racemic mixture of compounds can be carried out by any of many methods known in the art. One method is fractional recrystallization using a chiral resolving acid that is an optically active, salt-forming organic acid. Suitable resolving agents for fractional recrystallization are, for example, optically active acids, such as tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, or various optically active camphorsulfonic acids, such as the D and L forms of β-camphorsulfonic acid. Other suitable resolving agents for fractional crystallization include stereoisomerically pure forms of α-methyl-benzylamine (e.g., S and R forms, or diastereomerically pure forms), 2-phenylglycinol, norephedrine, ephedrine, N-methylephedrine, cyclohexylethylamine, 1,2-diaminocyclohexane, and the like.

[0037] Resolution of racemic mixtures may be accomplished by elution through a column packed with an optically active resolving agent (e.g., dinitrobenzoylphenylglycine). The composition of suitable elution solvents can be determined by one skilled in the art.

[0038] In some embodiments, the compounds of the invention have the (R) configuration. In other embodiments, the compounds have the (S) configuration. In compounds with more than one chiral center, each chiral center in the compound may be independently (R) or (S) unless otherwise indicated.

[0039] The compounds of the present invention also include tautomers. Tautomers result from the exchange of a single bond with an adjacent double bond along with the migration of a proton. Tautomers include prototropic tautomers, which are isomeric protonation states with the same empirical formula and total charge. Examples of prototropic tautomers include ketone-enol pairs, amide-imidic acid pairs, lactam-lactim pairs, enamine-imine pairs, and cyclic forms in which a proton can occupy more than one position in a heterocyclic ring system, such as 1H- and 3H-imidazole, 1H-, 2H- and 4H-1,2,4-triazole, 1H- and 2H-isoindole, and 1H- and 2H-pyrazole. Tautomers may be in equilibrium or sterically fixed in one form by appropriate substitution.

[0040] The compounds of the present invention may also include all isotopes of atoms occurring in intermediates or final compounds. Isotopes include atoms with the same atomic number but different mass numbers. For example, isotopes of hydrogen include tritium and deuterium. One or more constituent atoms of the compounds of the present invention may be replaced or substituted with an atomic isotope in natural or non-natural abundance ratio. In some embodiments, the compounds include at least one deuterium atom. For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced or substituted with deuterium. In some embodiments, the compounds include two or more deuterium atoms. In some embodiments, the compounds include 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 deuterium atoms. Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds can be used in a variety of studies, e.g., NMR spectroscopy, metabolic experiments, and / or assays.

[0041] Substitution with heavier isotopes, such as deuterium, may confer certain therapeutic advantages due to superior metabolic stability, e.g., increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances (A. Kerekes et.al. J. Med. Chem. 2011, 54, 201-210; R. Xu et.al. J. Label Compd. Radiopharm. 2015, 58, 308-312).

[0042] The term "compound" as used herein is meant to include all stereoisomers, geometric isomers, tautomers, and isotopes of the structures depicted. The term is also meant to refer to the compounds of the invention regardless of how they are prepared, for example, synthetically, through a biological process (e.g., metabolic or enzymatic transformation), or a combination thereof.

[0043] All compounds and their pharma- ceutically acceptable salts can be found together with other substances, such as water and solvents (e.g., hydrates and solvates) or can be isolated. When in the solid state, the compounds described herein and their salts can occur in various forms, for example, in the form of solvates, including hydrates. Compounds may be in any solid state form, such as polymorphs or solvates, and therefore, unless otherwise specified, references herein to compounds and their salts should be understood to include any solid state form of the compound.

[0044] In some embodiments, the compound of the present invention or its salt is substantially isolated. "Substantially isolated" means that the compound is at least partially or substantially separated from the environment in which it was formed or detected. Partial separation may include, for example, a composition enriched in the compound of the present invention. Substantial separation may include a composition that includes at least about 50% by weight, at least about 60% by weight, at least about 70% by weight, at least about 80% by weight, at least about 90% by weight, at least about 95% by weight, at least about 97% by weight, or at least about 99% by weight of the compound of the present invention or its salt.

[0045] The phrase "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions and / or dosage forms that are suitable, within the scope of sound medical judgment, for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0046] The expressions "ambient temperature" and "room temperature", as used herein, are understood in the art and generally refer to temperatures, e.g., reaction temperatures, which relate to the temperature of the room in which the reaction is carried out, e.g., a temperature of about 20° C. to about 30° C.

[0047] The present invention also includes pharma- ceutically acceptable salts of the compounds described herein. The term "pharma- ceutically acceptable salts" refers to derivatives of the disclosed compounds in which the parent compound is modified by converting an existing acid or base moiety into its salt form. Examples of pharma- ceutically acceptable salts include, but are not limited to, inorganic or organic acid salts of basic residues, such as amines; alkali or organic salts of acidic residues, such as carboxylic acids; and the like. The pharma- ceutical acceptable salts of the present invention include non-toxic salts of the parent compound formed, for example, from non-toxic inorganic or organic acids. The pharma- ceutical acceptable salts of the present invention may be synthesized from the parent compound containing a basic or acidic moiety by conventional chemical methods. In general, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or an organic solvent, or a mixture of the two; generally, non-aqueous media, such as ether, ethyl acetate, alcohols (e.g., methanol, ethanol, isopropanol, or butanol) or acetonitrile (MeCN) are preferred. A list of suitable salts can be found in Remington's Pharmaceutical Sciences 17 th Ed., (Mack Publishing Company, Easton, 1985), p. 1418, Berge et al., J. Pharm. Sci., 1977, 66(1), 1-19, and Stahl et al., Handbook of Pharmaceutical Salts: Properties, Selection, and Use, (Wiley, 2002). In some embodiments, the compounds described herein are in the N-oxide form.

[0048] synthesis The compounds of the invention, including their salts, may be prepared using known organic synthesis techniques and may be synthesised according to any of a number of possible synthetic routes, for example those in the following schemes.

[0049] The reaction for preparing the compounds of the present invention can be carried out in a suitable solvent, which can be easily selected by those skilled in the art of organic synthesis.A suitable solvent can be substantially non-reactive with the starting material (reactant), intermediate or product at the temperature at which the reaction is carried out, which can range, for example, from the freezing temperature of the solvent to the boiling temperature of the solvent.A given reaction can be carried out in one solvent or a mixture of more than one solvent.Depending on the specific reaction step, a suitable solvent for a specific reaction step can be selected by those skilled in the art.

[0050] Preparation of the compounds of the invention may involve the protection and deprotection of various chemical groups. The need for protection and deprotection, and the selection of appropriate protecting groups, can be readily determined by one of ordinary skill in the art. The chemistry of protecting groups can be found, for example, in Kocienski, Protecting Groups, (Thieme, 2007); Robertson, Protecting Group Chemistry, (Oxford University Press, 2000); Smith et al., March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 6 th Ed. (Wiley, 2007); Peturssion et al., “Protective Groups in Carbohydrate Chemistry,” J. Chem. Educ., 1997, 74(11), 1297; and Wuts et al., Protective Groups in Organic Synthesis, 4th Ed., (Wiley, 2006).

[0051] The reaction may be monitored according to any suitable method known in the art. For example, the formation of the product may be monitored by spectroscopic means, such as nuclear magnetic resonance spectroscopy (e.g., 1 H or 13C), infrared spectroscopy, spectrophotometry (eg, UV-visible), mass spectrometry, or by chromatographic methods such as high performance liquid chromatography (HPLC) or thin layer chromatography (TLC).

[0052] The following schemes provide general guidance relating to the preparation of compounds of the invention. Those skilled in the art will appreciate that the preparations shown in the schemes can be modified or optimized using general knowledge of organic chemistry to prepare various compounds of the invention.

[0053] [ka] Compounds of formula 1-18 can be prepared via the synthetic route outlined in Scheme 1. Starting material 1-1 can be halogenated with a suitable reagent (such as N-chloro-succinimide (NCS)) to give intermediate 1-2 (where Hal is a halide (such as F, Cl, Br, or I)). Compound 1-3 can be prepared by treating 1-2 with a reagent (such as triphosgene). Intermediate 1-3 can then be reacted with ester 1-4 to give nitro compound 1-5, which can be treated with a suitable reagent (e.g., POCl3) to give compound 1-6. S-coupling of intermediate 1-6 with amine 1-7 (where PG is a suitable protecting group (such as Boc)) can be performed. NAr reaction can be carried out to give compound 1-8. The nitro group in 1-8 can be reduced to NH2 in the presence of a reducing agent (e.g., Fe in acetic acid or sodium dithionite). Intermediate 1-9 can then be subjected to a cyclization reaction (e.g., using triethyl orthoformate) to give intermediate 1-10, followed by a SnAr reaction with sodium thiomethoxide to give 1-11. Cross-coupling reaction with 1-12 (M is a boronic acid, boronic ester, or appropriately substituted metal or metalloid [e.g., M is B(OR)2, Sn(alkyl)3, Zn-Hal, or CF3TMS]) under standard Suzuki cross-coupling conditions (e.g., in the presence of a palladium catalyst and a suitable base), or standard Stille cross-coupling conditions (e.g., in the presence of a palladium catalyst), or standard Negishi cross-coupling conditions (e.g., in the presence of a palladium catalyst), or trifluoromethylation conditions (e.g., in the presence of a copper catalyst) gives 1-13. Intermediate 1-15 can be prepared by cross-coupling reaction between 1-13 and an adduct of formula 1-14 (M is a boronic acid, a boronic ester, or an appropriately substituted metal [e.g., M is B(OR)2, Sn(alkyl)3, or Zn-Hal]) under standard Suzuki cross-coupling conditions (e.g., in the presence of a palladium catalyst and a suitable base), or standard Stille cross-coupling conditions (e.g., in the presence of a palladium catalyst), or standard Negishi cross-coupling conditions (e.g., in the presence of a palladium catalyst). Intermediate 1-15 can be converted to intermediate 1-16 via either oxidation of the sulfur group with a suitable oxidizing agent (e.g., m-CPBA) followed by a SnAr reaction, or a cross-coupling reaction (Org. Lett. 2002, 4, 979-981). Removal of the protecting group in 1-16 gives amine 1-17. Subsequent functionalization of the resulting amine, such as coupling with an acid chloride (e.g., acryloyl chloride), affords the desired products 1-18. The sequence of the above chemical reactions may be modified or omitted as necessary to suit the preparation of different analogs.

[0054] KRAS protein The Ras family consists of three members: KRAS, NRAS, and HRAS. RAS-mutated cancers account for approximately 25% of human cancers. KRAS is the most frequently mutated isoform in human cancers: 85% of all RAS mutations are in KRAS, 12% in NRAS, and 3% in HRAS (Simanshu, D. et al. Cell 170.1 (2017): 17-33). KRAS mutations are prevalent among the top three most lethal cancer types: pancreatic (97%), colorectal (44%), and lung (30%) (Cox, AD et al. Nat Rev Drug Discov (2014) 13: 828-51). The majority of RAS mutations occur at amino acid residues / codons 12, 13, and 61; mutations in codon 12 occur most frequently in KRAS. The frequency of certain mutations varies between RAS genes, with G12D mutations being the most prevalent in KRAS, while Q61R and G12R mutations occur most frequently in NRAS and HRAS. Furthermore, the spectrum of mutations in RAS isoforms differs between cancer types. For example, KRAS G12D mutations predominate in pancreatic cancer (51%), followed by colorectal adenocarcinoma (45%) and lung cancer (17%) (Cox, AD et al. Nat Rev Drug Discov (2014) 13:828-51). In contrast, KRAS G12C mutations predominate in non-small cell lung cancer (NSCLC) (where nearly half of mutant KRAS are G12C), accounting for 11-16% of lung adenocarcinomas and 2-5% of pancreatic and colorectal adenocarcinomas, respectively (Cox, AD et al. Nat. Rev. Drug Discov. (2014) 13:828-51a). Using shRNAs to knock down thousands of genes across hundreds of cancer cell lines, genomic studies have demonstrated that cancer cells exhibiting KRAS mutations are highly dependent on KRAS function to grow cells (McDonald, R. et al. Cell 170 (2017):577-592). Taken together, these findings suggest that KRAS mutations play a critical role in human cancers, and thus the development of inhibitors targeting mutant KRAS may be useful in the clinical treatment of diseases characterized by KRAS mutations.

[0055] How to use Cancer types involving KRAS, including G12C, G12V and G12D mutations, include, but are not limited to, adenocarcinomas (e.g., pancreatic, colorectal, lung, bladder, stomach, esophagus, breast, head and neck, neck skin, thyroid); hematopoietic malignancies (e.g., myeloproliferative neoplasms (MPN), myelodysplastic syndromes (MDS), chronic and juvenile myelomonocytic leukemia (CMML and JMML), acute myeloid leukemia (AML), acute lymphoblastic leukemia (ALL) and multiple myeloma (MM)); and other neoplasms (e.g., glioblastoma and sarcoma). Additionally, KRAS mutations have been associated with acquired resistance to anti-EGFR therapy (Knickelbein, K. et al. Genes & Cancer, (2015): 4-12). KRAS mutations have been found in immunological and inflammatory disorders (Fernandez-Medarde, A. et al. Genes & Cancer, (2011):344-358), such as Ras-associated lymphoproliferative disorder (RALD) or juvenile myelomonocytic leukemia (JMML), which are caused by somatic mutations in KRAS or NRAS.

[0056] The compound of the present disclosure can inhibit the activity of KRAS protein.For example, the compound of the present disclosure can be used to inhibit the activity of KRAS in cells or individuals or patients that require enzyme inhibition by administering to cells, individuals or patients an inhibitory amount of one or more compounds of the present disclosure.

[0057] As KRAS inhibitors, the compounds of the present disclosure are useful in treating various diseases associated with abnormal expression or activity of KRAS. Compounds that inhibit KRAS will be useful in providing a means of blocking growth or apoptosis induction in tumors, or by inhibiting angiogenesis. As a result, it is expected that the compounds of the present disclosure will prove useful in treating or blocking proliferative disorders, such as cancer. In particular, tumors with activated mutants of receptor tyrosine kinases or upregulation of receptor tyrosine kinases may be particularly sensitive to inhibitors.

[0058] In one aspect, provided herein is a method of inhibiting KRAS activity comprising contacting a compound of the present disclosure with KRAS. In some embodiments, the contacting comprises administering the compound to a patient.

[0059] In one aspect, provided herein is a method of inhibiting a KRAS protein having a G12C mutation, the method comprising contacting KRAS with a compound of the present disclosure.

[0060] In one aspect, provided herein is a method of inhibiting a KRAS protein having a G12D mutation, the method comprising contacting KRAS with a compound of the present disclosure.

[0061] In one aspect, provided herein is a method of inhibiting a KRAS protein having a G12V mutation, the method comprising contacting KRAS with a compound of the present disclosure.

[0062] In another aspect, provided herein is a method for treating a disease or disorder associated with inhibition of KRAS interaction, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any of the formulas disclosed herein, or a pharma- ceutically acceptable salt thereof.

[0063] In yet another aspect, the present specification provides a method for treating a disease or disorder associated with inhibition of a KRAS protein having a G12C mutation, comprising administering to a patient in need thereof a therapeutically effective amount of a compound of any of the formulas disclosed herein, or a pharma- ceutically acceptable salt thereof.

[0064] In yet another aspect, also provided herein is a method of treating cancer in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound disclosed herein, wherein the cancer is characterized by interaction with a KRAS protein having a G12C mutation.

[0065] In yet another aspect, provided herein is a method of treating cancer in a patient, comprising administering to the patient a therapeutically effective amount of any one of the compounds disclosed herein, or a pharma- ceutical acceptable salt thereof.

[0066] In another aspect, provided herein is a method for treating a disease or disorder associated with inhibition of KRAS interaction or a mutant thereof in a patient in need thereof, comprising administering to the patient a compound disclosed herein, or a pharma- ceutically acceptable salt thereof, or a composition comprising a compound disclosed herein, or a pharma- ceutically acceptable salt thereof, in combination with another therapy or treatment disclosed herein.

[0067] In one embodiment, the cancer is selected from hematological cancer, sarcoma, lung cancer, gastrointestinal cancer, genitourinary cancer, liver cancer, bone cancer, nervous system cancer, gynecological cancer, and skin cancer.

[0068] In another embodiment, the lung cancer is selected from non-small cell lung cancer (NSCLC), small cell lung carcinoma, bronchogenic carcinoma, squamous cell bronchogenic carcinoma, undifferentiated small cell bronchogenic carcinoma, undifferentiated large cell bronchogenic carcinoma, adenocarcinoma, bronchogenic carcinoma, alveolar carcinoma, bronchiolar carcinoma, bronchial adenoma, chondromatous hamartoma, mesothelioma, papillary and nonpapillary carcinoma, bronchial adenoma, and pleuropulmonary blastoma.

[0069] In yet another embodiment, the lung cancer is non-small cell lung cancer (NSCLC).In yet another embodiment, the lung cancer is adenocarcinoma.

[0070] In one embodiment, the gastrointestinal cancer is selected from esophageal squamous cell carcinoma, esophageal adenocarcinoma, esophageal leiomyosarcoma, esophageal lymphoma, gastric carcinoma, gastric lymphoma, gastric leiomyosarcoma, exocrine pancreatic carcinoma, pancreatic ductal adenocarcinoma, pancreatic insulinoma, pancreatic glucagonoma, pancreatic gastrinoma, pancreatic carcinoid tumor, pancreatic vipoma, small intestinal adenocarcinoma, small intestinal lymphoma, small intestinal carcinoid tumor, Kaposi's sarcoma, small intestinal leiomyoma, small intestinal hemangioma, small intestinal lipoma, small intestinal neurofibroma, small intestinal fibroma, large intestinal adenocarcinoma, large intestinal tubular adenoma, large intestinal villous adenoma, large intestinal hamartoma, large intestinal leiomyoma, colorectal carcinoma, gallbladder carcinoma, and anal carcinoma.

[0071] In one embodiment, the gastrointestinal cancer is colorectal cancer.

[0072] In another embodiment, the cancer is a carcinoma. In yet another embodiment, the carcinoma is selected from pancreatic carcinoma, colorectal carcinoma, lung carcinoma, bladder carcinoma, gastric carcinoma, esophageal carcinoma, breast carcinoma, head and neck carcinoma, cervical skin carcinoma, and thyroid carcinoma.

[0073] In yet another embodiment, the cancer is a hematopoietic malignancy. In one embodiment, the hematopoietic malignancy is selected from multiple myeloma, acute myeloid leukemia, and myeloproliferative neoplasms.

[0074] In another embodiment, the cancer is a neoplasm. In yet another embodiment, the neoplasm is a glioblastoma or a sarcoma.

[0075] In certain embodiments, the present disclosure provides a method for treating a KRAS-mediated disorder in a patient in need thereof, comprising administering to the patient a compound according to the present invention or a pharma- ceutical acceptable composition thereof.

[0076] In some embodiments, diseases and indications treatable using the compounds of the present disclosure include, but are not limited to, hematological cancers, sarcomas, lung cancer, gastrointestinal cancer, genitourinary cancer, liver cancer, bone cancer, nervous system cancer, gynecological cancer, and skin cancer.

[0077] Exemplary hematological cancers include lymphomas and leukemias, such as acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin's lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin's lymphoma, myeloproliferative disorders, and myeloproliferative disorders. proliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), essential thrombocytosis (ET), 8p11 myeloproliferative syndrome, myelodysplastic syndromes (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), multiple myeloma, cutaneous T-cell lymphoma, adult T-cell leukemia, Waldenstrom's macroglubulinemia, hairy cell lymphoma, marginal zone lymphoma, chronic myelogenous lymphoma, and Burkitt's lymphoma).

[0078] Exemplary sarcomas include chondrosarcoma, Ewing's sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxoma, rhabdomyoma, rhabdomyosarcoma, fibroma, lipoma, hamartoma, lymphosarcoma, leiomyosarcoma, and teratoma.

[0079] Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung carcinoma, bronchogenic carcinoma (squamous cell, small undifferentiated cell, large undifferentiated cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, chondromatous hamartoma, mesothelioma, papillary and nonpapillary carcinoma, bronchial adenoma, and pleuropulmonary blastoma.

[0080] Exemplary gastrointestinal cancers include esophageal cancer (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), gastric cancer (carcinoma, lymphoma, leiomyosarcoma), pancreatic cancer (exocrine pancreatic carcinoma, ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, vipoma), small intestine cancer (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi's sarcoma, leiomyoma, hemangioma, lipoma, neurofibroma, fibroma), colon cancer (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyoma), colorectal cancer, gallbladder cancer, and anal cancer.

[0081] Exemplary genitourinary cancers include renal carcinoma (adenocarcinoma, Wilms' tumor [nephroblastoma], renal cell carcinoma), bladder and urethral carcinoma (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate carcinoma (adenocarcinoma, sarcoma), testicular carcinoma (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, stromal cell carcinoma, fibroma, fibroadenoma, adenomatous tumor, lipoma), and urothelial carcinoma.

[0082] Exemplary liver cancers include hepatocarcinoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma, and hemangioma.

[0083] Exemplary bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulum cell sarcoma), multiple myeloma, malignant giant cell tumor chordoma, osteochondral osteoma (osteochondral exostosis), benign chondroma, chondroblastoma, chondromyxoid fibroma, osteoid osteoma, and giant cell tumor.

[0084] Exemplary nervous system cancers include cancer of the skull (osteoma, hemangioma, granuloma, xanthomas, osteitis deformans), meninges (meningioma, meningeal sarcoma, glioma), brain (astrocytoma, mesodermoma, glioma, ependymoma, germinoma (pinealoma), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumors, neuroectodermal tumors), and spinal cord (neurofibroma, meningioma, glioma, sarcoma), neuroblastoma, Lhermitte-Duclos disease, and pineal tumor.

[0085] Exemplary gynecological cancers include breast (ductal carcinoma, lobular carcinoma, breast sarcoma, triple-negative breast cancer, HER2-positive breast cancer, inflammatory breast cancer, papillary carcinoma), uterus (endometrial carcinoma), cervix, Cancers of the vagina (cervical carcinoma, preneoplastic cervical dysplasia), ovary (ovarian carcinoma (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa cell tumor, Sertoli-Leydig cell tumor, dysgerminoma, malignant teratoma), vulva (squamous cell carcinoma, carcinoma in situ, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, botryoid sarcoma (embryonic rhabdomyosarcoma)), and fallopian tube (carcinoma).

[0086] Exemplary skin cancers include melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, Merkel cell skin cancer, moles dysplastic nevi, lipoma, hemangioma, dermatofibroma, and keloids.

[0087] Exemplary head and neck cancers include glioblastoma, melanoma, rhabdomyosarcoma, lymphosarcoma, osteosarcoma, squamous cell adenocarcinoma, adenocarcinoma, oral cancer, laryngeal cancer, nasopharyngeal cancer, nasal and paranasal sinus cancer, thyroid and parathyroid cancer, eye tumors, lip and mouth tumors, and squamous head and neck cancer.

[0088] Compounds of the present disclosure may also be useful in inhibiting tumor metastasis.

[0089] In addition to oncogenic neoplasms, the compounds of the present invention are useful in the treatment of skeletal and chondrocyte disorders, including, but not limited to, achondroplasia, hypochondroplasia, dwarfism, totodile dysplasia (TD) (TDI and TDII clinical types), Apert syndrome, Crouzon syndrome, Jackson-Weiss syndrome, Bear-Stevenson cutis rotation syndrome, Pfeiffer syndrome, and craniosynostosis syndrome. In some embodiments, the present disclosure provides methods for treating patients suffering from skeletal and chondrocyte disorders.

[0090] In some embodiments, the compounds described herein can be used to treat Alzheimer's disease, HIV, or tuberculosis.

[0091] As used herein, the term "8p11 myeloproliferative syndrome" is meant to refer to myeloid / lymphoid neoplasms associated with hypereosinophilia and FGFR1 abnormalities.

[0092] As used herein, the term "cell" is meant to refer to a cell that exists in vitro, ex vivo, or in vivo. In some embodiments, an ex vivo cell may be part of a tissue sample excised from an organism, e.g., a mammal. In some embodiments, an in vitro cell may be a cell in cell culture. In some embodiments, an in vivo cell is a cell that is living within an organism, e.g., a mammal.

[0093] As used herein, the term "contacting" refers to bringing together the indicated moieties in an in vitro system or in vivo system.For example, "contacting" KRAS with the compound described herein includes administering the compound described herein to an individual or patient, such as a human, that has KRAS, as well as introducing the sample, such as the cell preparation or purified preparation that contains KRAS, into the compound described herein.

[0094] As used herein, the terms "individual," "subject," or "patient" are used interchangeably and refer to any animal, including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cows, sheep, horses, or primates, most preferably humans.

[0095] As used herein, the phrase "therapeutically effective amount" refers to an amount of an active compound or pharmaceutical agent, such as any of the solid forms or salts thereof disclosed herein, that elicits the biological or medicinal response in a tissue, system, animal, individual or human that is desired by a researcher, veterinarian, physician or other clinician. An appropriate "effective" amount in any particular case may be determined using techniques known to those of ordinary skill in the art.

[0096] The phrase "pharmacologically acceptable" is used herein to refer to compounds, materials, compositions, and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals and free of undue toxicity, irritation, allergic response, immunogenicity or other problem or complication commensurate with a reasonable benefit / risk ratio.

[0097] As used herein, the phrase "pharmaceutically acceptable carrier or excipient" refers to a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid excipient, diluent, solvent, or encapsulating material. Excipients or carriers are generally safe, non-toxic, and not otherwise biologically undesirable, and include excipients or carriers that are acceptable for veterinary use as well as human pharmaceutical use. In one embodiment, each component is "pharmaceutically acceptable" as defined herein. For example, Remington:The Science and Practice of Pharmacy,21st ed.;Lippincott Williams & Wilkins:Philadelphia,Pa.,2005;Handbook of Pharmaceutical Excipients,6th ed.;Rowe et al.,Eds.;The Pharmaceutical Press and the American Pharmaceutical Association:2009;Handbook of Pharmaceutical Additives,3rd ed.;Ash and Ash Eds.;Gower Publishing Company:2007;Pharmaceutical Preformulation and Formulation, 2nd ed.;Gibson Ed.;CRC Press LLC:Boca Raton,Fla.,2009.

[0098] As used herein, the term "treating" or "treatment" refers to inhibiting a disease; e.g., inhibiting a disease, condition, or disorder in an individual experiencing or exhibiting a symptom or symptom of the disease, condition, or disorder (i.e., halting further progression of the symptom and / or symptom), or ameliorating a disease; e.g., ameliorating a disease, condition, or disorder in an individual experiencing or exhibiting a symptom or symptom of the disease, condition, or disorder (i.e., reversing the symptom and / or symptom), e.g., reducing the severity of a disease.

[0099] The terms "prevent," "preventing," or "prevention," as used herein, include preventing at least one symptom associated with or caused by the condition, disease, or disorder being prevented.

[0100] It is appreciated that certain features of the invention, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment (and it is intended that the embodiments be combined as if described in a multiple sub-form). Conversely, various features of the invention, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable subcombination.

[0101] Combination therapy I. Cancer Therapy Cancer cell growth and survival may be affected by the failure of multiple signaling pathways. Therefore, combining inhibitors of different enzymes / proteins / receptors that exhibit different selectivity for the targets they modulate is useful for treating such conditions. Targeting multiple signaling pathways (or multiple biological molecules involved in a given signaling pathway) can reduce the possibility of drug resistance occurring in cell populations and / or mitigate the toxicity of treatment.

[0102] One or more additional pharmaceutical agents, such as chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressants, immuno-oncology agents, metabolic enzyme inhibitors, chemokine receptor inhibitors, and phosphatase inhibitors, as well as targeted therapies, such as Bcr-Abl, Flt-3, EGFR, HER2, JAK, c-MET, VEGFR, PDGFR, c-Kit, IGF-1R, RAF, FAK, and CDK4 / 6 kinase inhibitors, such as those described in WO2006 / 056399, can be used in combination with the compounds of the present disclosure to treat a CDK2-related disease, disorder, or condition. Other agents, such as therapeutic antibodies, can be used in combination with the compounds of the present disclosure to treat a CDK2-related disease, disorder, or condition. One or more additional pharmaceutical agents can be administered to the patient simultaneously or sequentially.

[0103] In some embodiments, a CDK2 inhibitor is administered or used in combination with a BCL2 inhibitor or a CDK4 / 6 inhibitor.

[0104] The compounds disclosed herein can be combined with one or more other enzyme / protein / receptor inhibitor therapies for the treatment of diseases such as cancer and other diseases or disorders described herein. Examples of diseases and indications treatable with combination therapy include those described herein. Examples of cancer include solid tumors and non-solid tumors such as liquid tumors, hematological cancers, etc. Examples of infectious diseases include viral infections, bacterial infections, fungal infections, or parasitic infections. For example, the compounds disclosed herein can be combined with one or more inhibitors of the following kinases for the treatment of cancer: Akt1, Akt2, Akt3, BCL2, CDK4 / 6, TGF-βR, PKA, PKG, PKC, CaM kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IDH2, IGF-1R, IR-R, PDGFαR, PDGFβR, PI3K (alpha, beta, gamma, delta, and multiple or selective), CSF1R, KIT, FLK-II, KDR / FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, PARP, Ron, Sea, TRKA, TRKB, TRKC, TAM kinases (Axl, Mer, Tyro3), FLT3, VEGFR / Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK, and B-Raf. In some embodiments, the compounds of the present disclosure can be combined with one or more of the following inhibitors for the treatment of cancer or infectious diseases: Non-limiting examples of inhibitors that can be combined with the compounds of the present disclosure for the treatment of cancer and infectious diseases include FGFR inhibitors (FGFR1, FGFR2, FGFR3, or FGFR4, e.g., pemigatinib (INCB54828), INCB62079), EGFR inhibitors (also known as ErB-1 or HER-1;e.g., erlotinib, gefitinib, vandetanib, orsimertinib, cetuximab, necitumumab, or panitumumab), VEGFR inhibitors or pathway blockers (e.g., bevacizumab, pazopanib, sunitinib, sorafenib, axitinib, regorafenib, ponatinib, cabozantinib, vandetanib, ramucirumab, lenvatinib, ziv-aflibercept), PARP inhibitors (e.g., e.g., olaparib, rucaparib, veliparib, or niraparib), JAK inhibitors (JAK1 and / or JAK2, e.g., ruxolitinib or baricitinib; or JAK1, e.g., itacitinib (INCB39110), INCB052793, or INCB054707), IDO inhibitors (e.g., epacadostat, NLG919, or BMS-986205, MK7162), LSD1 inhibitors (e.g., GSK2979552, INCB59872, and INCB60003), TDO inhibitors, PI3K-delta inhibitors (e.g., palsaclisib (INCB50465) or INCB50797), PI3K-gamma inhibitors such as PI3K-gamma selective inhibitors, Pim inhibitors (e.g., INCB53914), CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl, and Mer; e.g., INCB081776), adenosine receptor antagonists (e.g., A2a / A2b receptor antagonists), HPK1 inhibitors, chemokine receptor inhibitors (e.g., CCR2 or CCR5 inhibitors), SHP1 / 2 phosphatase inhibitors, histone deacetylase inhibitors (HDACs) such as HDAC8 inhibitors, angiogenesis inhibitors, interleukin receptor inhibitors, bromo and extra terminal family member inhibitors (e.g., bromodomain inhibitors or BET inhibitors, such as INCB54329 and INCB57643), c-MET inhibitors (e.g., capmatinib), anti-CD19 antibodies (e.g., tafasitamab), ALK2 inhibitors (e.g., INCB00928); or combinations thereof.

[0105] In some embodiments, the compounds or salts described herein are administered with a PI3Kδ inhibitor. In some embodiments, the compounds or salts described herein are administered with a JAK inhibitor. In some embodiments, the compounds or salts described herein are administered with a JAK1 or JAK2 inhibitor (e.g., baricitinib or ruxolitinib). In some embodiments, the compounds or salts described herein are administered with a JAK1 inhibitor. In some embodiments, the compounds or salts described herein are administered with a JAK1 inhibitor that is more selective than JAK2.

[0106] In addition, the compounds described herein can be used in combination with targeted therapies, such as c-MET inhibitors (e.g., capmatinib), anti-CD19 antibodies (e.g., tafasitamab), ALK2 inhibitors (e.g., INCB00928), or combinations thereof, to treat cancer and other proliferative diseases.

[0107] Examples of antibodies used in combination therapy include, but are not limited to, trastuzumab (e.g., anti-HER2), ranibizumab (e.g., anti-VEGF-A), bevacizumab (Avastin™, e.g., anti-VEGF), panitumumab (e.g., anti-EGFR), cetuximab (e.g., anti-EGFR), Rituxan (e.g., anti-CD20), and antibodies against c-MET.

[0108] One or more of the following agents may be used in combination with the compounds of the present disclosure, which are presented as a non-limiting list: cytostatic agents, cisplatin, doxorubicin, taxotere, taxol, etoposide, irinotecan, camptostar, topotecan, paclitaxel, docetaxel, epothilone, tamoxifen, 5-fluorouracil, methotrexate, temozolomide, cyclophosphamide, SCH 66336, R115777, L778,123, BMS 214662, IRESSA™ (gefitinib), TARCEVA™ (erlotinib), antibodies against EGFR, intron, ara-C, adriamycin, cytoxan, gemcitabine, uracil mustard, chlormethine, ifosfamide, melphalan, chlorambucil, pipobroman, triethylenemelamine, triethylenethiophosphoramine, busulfan, carmustine, lomustine, streptozocin, dacarbazine, floxuridine, cyta Rabin, 6-mercaptopurine, 6-thioguanine, fludarabine phosphate, oxaliplatin, leucovorin, ELOXATIN™ (oxaliplatin), pentostatin, vinblastine, vincristine, vindesine, bleomycin, dactinomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mithramycin, deoxycoformycin, mitomycin-C, L-asparaginase, teniposide 17.Alpha-ethynyl estradiol, diethylstilbestrol, testosterone, prednisone, fluoxymesterone, dromostanolone propionate, testolactone, megestrol acetate, methylprednisolone, methyltestosterone, prednisolone, triamcinolone, chlorotrianisene, hydroxyprogesterone, aminoglutethimide, estramustine, medroxyprogesterone acetate, leuprolide, flutamide, toremifene, goserelin, carboplatin, hydroxyurea, amsacrine, procarbazine, mitotane, mitoxantrone, levamisole, navelbine, anastrozole, letrazole, capecitabine, reloxafine, droloxafine, hexamethylmelamine, avastin, HERCEPTIN™ (trastuzumab ), BEXXAR™ (tositumomab), VELCADE™ (bortezomib), ZEVALIN™ (ibritumomab tiuxetan), TRISENOX™ (arsenic trioxide), XELODA™ (capecitabine), vinorelbine, porfimer, ERBITUX™ (cetuximab), thiotepa, altretamine, melphalan, trastuzumab, relozole, Fulvestrant, Exemestane, Ifosfamide, Rituximab, C225 (Cetuximab), Campath (Alemtuzumab), Clofarabine, Cladribine, Aphidicolin, Rituxan, Sunitinib, Dasatinib, Tezacitabine, Sml1, Fludarabine, Pentostatin, Triapine, Didox, Trimidox, Amidox, 3-AP, and MDL-101,731.

[0109] The compounds of the present disclosure can also be combined with other cancer treatment methods, such as chemotherapy, radiation therapy, tumor-targeted therapy, adjuvant therapy, immunotherapy, or surgery. Examples of immunotherapy include cytokine therapy (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, bispecific or multispecific antibodies, antibody-drug conjugates, T-cell adoptive transfer, Toll receptor agonists, RIG-I agonists, oncolytic virus therapy, and immunomodulatory small molecules, including thalidomide or JAK1 / 2 inhibitors, PI3Kδ inhibitors, and the like. The compounds can be administered in combination with one or more anti-cancer agents, such as chemotherapeutic agents. Examples of chemotherapy agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, intravenous busulfan, oral busulfan, calsterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, drostanolone propionate, and eculizumab. , epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon alpha 2a, irinotecan, lapatinib tosylate hydrate, lenalidomide, letrozole, leucovorin, leuprorelin acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone,Nandrolone phenpropionate phenpropionate), nelarabine, nofetumomab, oxaliplatin, paclitaxel, pamidronic acid, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, ruxolitinib, sorafenib, streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan, toremifene, tositumomab, trastuzumab, tretinoin, uracil mustard, valrubicin, vinblastine, vincristine, vinorelbine, vorinostat, and zoledronate.

[0110] Additional examples of chemotherapeutic agents include proteosome inhibitors (eg, bortezomib), thalidomide, revlimid, and DNA damaging agents such as melphalan, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

[0111] Examples of steroids include corticosteroids such as dexamethasone or prednisone.

[0112] Examples of Bcr-Abl inhibitors include imatinib mesylate (GLEEVAC™), nilotinib, dasatinib, bosutinib, and ponatinib, and pharmaceutically acceptable salts. Other suitable examples of Bcr-Abl inhibitors include the genera and species of compounds disclosed in U.S. Patent No. 5,521,184, WO04 / 005281, and U.S. Application No. 60 / 578,491, and pharmaceutically acceptable salts thereof.

[0113] Examples of suitable Flt-3 inhibitors include midostaurin, lestaurtinib, linifanib, sunitinib, sunitinib, maleate, sorafenib, quizartinib, crenolanib, pacritinib, tanzutinib, PLX3397, and ASP2215, and pharmaceutically acceptable salts thereof. Examples of other suitable Flt-3 inhibitors include the compounds disclosed in WO03 / 037347, WO03 / 099771, and WO04 / 046120, and pharmaceutically acceptable salts thereof.

[0114] Examples of suitable RAF inhibitors include dabrafenib, sorafenib, and vemurafenib, and pharmaceutically acceptable salts thereof. Other suitable RAF inhibitors include the compounds disclosed in WO00 / 09495 and WO05 / 028444, and pharmaceutically acceptable salts thereof.

[0115] Examples of suitable FAK inhibitors include VS-4718, VS-5095, VS-6062, VS-6063, BI853520, and GSK2256098, and pharma- ceutically acceptable salts thereof. Other examples of suitable FAK inhibitors include the compounds disclosed in WO04 / 080980, WO04 / 056786, WO03 / 024967, WO01 / 064655, WO00 / 053595, and WO01 / 014402, and pharma- ceutically acceptable salts thereof.

[0116] Examples of suitable CDK4 / 6 inhibitors include palbociclib, ribociclib, trilaciclib, relociclib, and abemaciclib, and pharmaceutically acceptable salts thereof. Examples of other suitable CDK4 / 6 inhibitors include the compounds disclosed in WO09 / 085185, WO12 / 129344, WO11 / 101409, WO03 / 062236, WO10 / 075074, and WO12 / 061156, and pharmaceutically acceptable salts thereof.

[0117] In some embodiments, the compounds of the present disclosure can be used in combination with one or more other kinase inhibitors, including imatinib, particularly for treating patients who are resistant to imatinib or other kinase inhibitors.

[0118] In some embodiments, the compounds of the present disclosure can be used in combination with chemotherapeutic agents in the treatment of cancer, and can improve the therapeutic response without worsening the toxic effects of the chemotherapeutic agent compared to the response to the chemotherapeutic agent alone. In some embodiments, the compounds of the present disclosure can be used in combination with chemotherapeutic agents as set forth herein. For example, additional pharmaceutical agents used in the treatment of multiple myeloma can include, but are not limited to, melphalan, melphalan + prednisone [MP], doxorubicin, dexamethasone, and Velcade (bortezomib). Further additional agents used in the treatment of multiple myeloma include Bcr-Abl, Flt-3, RAF, and FAK kinase inhibitors. In some embodiments, the agent is an alkylating agent, a proteasome inhibitor, a corticosteroid, or an immunomodulatory agent. Examples of alkylating agents include cyclophosphamide (CY), melphalan (MEL), and bendamustine. In some embodiments, the proteasome inhibitor is carfilzomib. In some embodiments, the corticosteroid is dexamethasone (DEX). In some embodiments, the immunomodulatory agent is lenalidomide (LEN) or pomalidomide (POM). A desirable outcome of the combination of the CDK2 inhibitors of the present disclosure with an additional agent is an additive or synergistic effect.

[0119] The agents may be combined with the compounds of the invention in a single or continuous release dosage form, or each agent may be administered simultaneously or sequentially in separate dosage forms.

[0120] The compounds of the present disclosure can be used in combination with one or more other inhibitors or one or more therapies for the treatment of infectious diseases, including viral, bacterial, fungal, or parasitic infections.

[0121] In some embodiments, a corticosteroid, such as dexamethasone, is administered to the patient in combination with a compound of the present disclosure, where the dexamethasone is administered intermittently rather than continuously.

[0122] The compounds of formula (I) or any of the formulas described herein, the compounds listed in any of the claims and described herein, or salts thereof, can be combined with other immunogenic agents, such as cancer cells, purified tumor antigens (including recombinant proteins, peptides, and carbohydrate molecules), cells, and cells transfected with genes encoding immune stimulating cytokines. Non-limiting examples of tumor vaccines that can be used include transfected tumor cells expressing melanoma antigens, such as peptides of gp100, MAGE antigens, Trp-2, MARTI, and / or tyrosinase, or the cytokine GM-CSF.

[0123] For cancer treatment, the compounds of formula (I) or any of the formulas described herein, the compounds listed in any of the claims and described herein, or salts thereof, can be combined with vaccination protocols. In some embodiments, tumor cells are transduced to express GM-CSF. In some embodiments, tumor vaccines include proteins from viruses involved in human cancers, such as human papillomavirus (HPV), hepatitis viruses (HBV and HCV), and Kaposi's herpes sarcoma virus (KHSV). In some embodiments, the compounds of the present disclosure can be combined with tumor-specific antigens, such as heat shock proteins isolated from the tumor tissue itself. In some embodiments, the compounds of formula (I) or any of the formulas described herein, the compounds listed in any of the claims and described herein, or salts thereof, can be combined with dendritic cell immunotherapy to activate a strong anti-tumor response.

[0124] The compounds of the present disclosure can be combined with bispecific macrocyclic peptides that target Fe alpha receptor or Fe gamma receptor expressing effector cells to tumor cells. The compounds of the present disclosure can also be combined with macrocyclic peptides that activate host immune responsiveness.

[0125] In some further embodiments, the combination of the disclosed compounds with other therapeutic agents can be administered to the patient prior to, during, and / or after bone marrow or stem cell transplantation. The disclosed compounds can be used in combination with bone marrow transplantation for the treatment of various tumors of hematopoietic origin.

[0126] The compounds of formula (I) or any of the formulas described herein, the compounds listed in any of the claims and described herein, or their salts can be used in combination with vaccines to stimulate immune responses against pathogens, toxins, and self-antigens. Examples of pathogens for which this therapeutic approach may be particularly useful include those for which there is currently no effective vaccine or for which traditional vaccines are not completely effective. Such pathogens include, but are not limited to, HIV, hepatitis (A, B, C), influenza, herpes, giardia, malaria, leishmania, Staphylococcus aureus, and Pseudomonas Aeruginosa.

[0127] Viruses causing infectious diseases treatable by the methods of the present disclosure include, but are not limited to, human papillomavirus, influenza, hepatitis A, B, C or D virus, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory syndrome virus, Ebola virus, measles virus, herpes virus (e.g., VZV, HSV-1, HAV-6, HSV-II, and CMV, Epstein-Barr virus), flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, soft wart virus, poliovirus, rabies virus, JC virus, and arboviral encephalitis virus.

[0128] Pathogens causing infections treatable by the methods of the present disclosure include, but are not limited to, Chlamydia, Rickettsia, Mycobacteria, Staphylococcus, Streptococcus, Pneumococcus, Neisseria meningitidis and Conococcus, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Corynebacterium diphtheriae, Salmonella, Bacillus, Vibrio cholera, Clostridium tetani, Clostridium botulinum, Bacillus anthracis, Plague, Leptospira, and Lyme burgdorferi.

[0129] Pathogenic fungi causing infections treatable by the methods of the present disclosure include, but are not limited to, Candida (e.g., albicans, krusei, glabrata, tropicalis), Cryptococcus neoformans, Aspergillus (e.g., fumigatus, niger), Genus Mucorales (e.g., mucor, absidia, rhizophus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, and Histoplasma capsulatum.

[0130] Pathogenic parasites causing infections treatable by the methods of the present disclosure include, but are not limited to, Entamoeba histolytica, Balantidium coli, Naegleria fowleri, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, and Nippostrongylus brasiliensis.

[0131] When multiple pharmaceutical agents are administered to a patient, they may be administered simultaneously, separately, sequentially, or in combination (eg, involving two or more agents).

[0132] Methods for safely and effectively administering most of these chemotherapeutic agents are known to those skilled in the art. In addition, their administration is described in standard texts. For example, the administration of many chemotherapeutic agents is described in the "Physicians' Desk Reference" (PDR, e.g., 1996 edition, Medical Economics Company, Montvale, NJ), the disclosure of which is incorporated herein by reference as if fully set forth.

[0133] II. Immune checkpoint therapy For the treatment of diseases such as cancer or infectious diseases, the compounds of the present disclosure can be used in combination with one or more immune checkpoint inhibitors. Examples of immune checkpoint inhibitors include inhibitors against immune checkpoint molecules such as CBL-B, CD20, CD28, CD40, CD70, CD122, CD96, CD73, CD47, CDK2, GITR, CSF1R, JAK, PI3K delta, PI3K gamma, TAM, arginase, HPK1, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, TLR (TLR7 / 8), TIGIT, CD112R, VISTA, PD-1, PD-L1, and PD-L2. In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In some embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, TIGIT, and VISTA. In some embodiments, the compounds provided herein can be used in combination with one or more agents selected from a KIR inhibitor, a TIGIT inhibitor, a LAIR1 inhibitor, a CD160 inhibitor, a 2B4 inhibitor, and a TGFR beta inhibitor.

[0134] In some embodiments, the compounds provided herein can be used in combination with one or more agonists of immune checkpoint molecules, such as OX40, CD27, GITR, and CD137 (also known as 4-1BB).

[0135] In some embodiments, the inhibitor of an immune checkpoint molecule is an anti-PD1 antibody, an anti-PD-L1 antibody, or an anti-CTLA-4 antibody.

[0136] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-1 or PD-L1, e.g., an anti-PD-1 or anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-1 or anti-PD-L1 antibody is selected from the group consisting of nivolumab, pembrolizumab, atezolizumab, durvalumab, avelumab, cemiplimab, atezolizumab, avelumab, tislelizumab, spartalizumab (PDR001), cetrelimab (JNJ-63723283), toripalimab (JS001), camrelizumab (SHR-1210), sintilimab (IBI308), AB122 (GLS-010), AMP-224, AMP-514 / MEDI -0680, BMS936559, JTX-4014, BGB-108, SHR-1210, MEDI4736, FAZ053, BCD-100, KN035, CS1001, BAT1306, LZM009, AK105, HLX10, SHR-1316, CBT-502(TQB2450), A167(KL-A167), STI-A101(ZKAB001), CK-301, BGB-A333, MSB-2311, HLX20, TSR-042, or LY3300054.In some embodiments, the PD-1 or PD-L1 inhibitor is a PD-L1 inhibitor as described in U.S. Patent Nos. 7,488,802, 7,943,743, 8,008,449, 8,168,757, 8,217,149, or 10,308,644; U.S. Publication Nos. 2017 / 0145025, 2017 / 0174671, 2017 / 0174679, No. 2017 / 0320875, No. 2017 / 0342060, No. 2017 / 0362253, No. 2018 / 0016260, No. 2018 / 0057486, No. 201 8 / 0177784, 2018 / 0177870, 2018 / 0179179, 2018 / 0179201, 2018 / 0179202, 2018 / 02 73519, 2019 / 0040082, 2019 / 0062345, 2019 / 0071439, 2019 / 0127467, 2019 / 0144439, 2019 / 0202824, 2019 / 0225601, 2019 / 0300524, or 2019 / 0345170; or PCT Publication No. WO03 042402, WO2008156712, WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400, or WO2011161699, each of which is incorporated by reference in its entirety. In some embodiments, the PD-L1 inhibitor is INCB086550.

[0137] In some embodiments, the antibody is an anti-PD-1 antibody, such as an anti-PD-1 monoclonal antibody. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, cemiplimab, spartalizumab, camrelizumab, cetrelimab, toripalimab, sintilimab, AB122, AMP-224, JTX-4014, BGB-108, BCD-100, BAT1306, LZM009, AK105, HLX10, or TSR-042. In some embodiments, the anti-PD-1 antibody is nivolumab, pembrolizumab, cemiplimab, spartalizumab, camrelizumab, cetrelimab, toripalimab, or sintilimab. In some embodiments, the anti-PD-1 antibody is pembrolizumab. In some embodiments, the anti-PD-1 antibody is nivolumab. In some embodiments, the anti-PD-1 antibody is cemiplimab. In some embodiments, the anti-PD-1 antibody is spartalizumab. In some embodiments, the anti-PD-1 antibody is camrelizumab. In some embodiments, the anti-PD-1 antibody is cetrelimab. In some embodiments, the anti-PD-1 antibody is toripalimab. In some embodiments, the anti-PD-1 antibody is sintilimab. In some embodiments, the anti-PD-1 antibody is AB122. In some embodiments, the anti-PD-1 antibody is AMP-224. In some embodiments, the anti-PD-1 antibody is JTX-4014. In some embodiments, the anti-PD-1 antibody is BGB-108. In some embodiments, the anti-PD-1 antibody is BCD-100. In some embodiments, the anti-PD-1 antibody is BAT1306. In some embodiments, the anti-PD-1 antibody is LZM009. In some embodiments, the anti-PD-1 antibody is AK105. In some embodiments, the anti-PD-1 antibody is HLX10. In some embodiments, the anti-PD-1 antibody is TSR-042. In some embodiments, the anti-PD-1 monoclonal antibody is nivolumab or pembrolizumab. In some embodiments, the anti-PD-1 monoclonal antibody is MGA012 (INCMGA0012; retifanlimab). In some embodiments, the anti-PD1 antibody is SHR-1210.Other anti-cancer agent(s) include antibody therapeutics such as 4-1BB (e.g., urelumab, utomirumab). In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PD-L1, e.g., an anti-PD-L1 monoclonal antibody. In some embodiments, the anti-PD-L1 monoclonal antibody is atezolizumab, avelumab, durvalumab, tislelizumab, BMS-935559, MEDI4736, atezolizumab (MPDL3280A; also known as RG7446), avelumab (MSB0010718C), FAZ053, KN035, CS1001, SHR-1316, CBT-502, A167, STI-A101, CK-301, BGB-A333, MSB-2311, HLX20, or LY3300054. In some embodiments, the anti-PD-L1 antibody is atezolizumab, avelumab, durvalumab, or tislelizumab. In some embodiments, the anti-PD-L1 antibody is atezolizumab. In some embodiments, the anti-PD-L1 antibody is avelumab. In some embodiments, the anti-PD-L1 antibody is durvalumab. In some embodiments, the anti-PD-L1 antibody is tislelizumab. In some embodiments, the anti-PD-L1 antibody is BMS-935559. In some embodiments, the anti-PD-L1 antibody is MEDI4736. In some embodiments, the anti-PD-L1 antibody is FAZ053. In some embodiments, the anti-PD-L1 antibody is KN035. In some embodiments, the anti-PD-L1 antibody is CS1001. In some embodiments, the anti-PD-L1 antibody is SHR-1316. In some embodiments, the anti-PD-L1 antibody is CBT-502. In some embodiments, the anti-PD-L1 antibody is A167. In some embodiments, the anti-PD-L1 antibody is STI-A101. In some embodiments, the anti-PD-L1 antibody is CK-301. In some embodiments, the anti-PD-L1 antibody is BGB-A333. In some embodiments, the anti-PD-L1 antibody is MSB-2311. In some embodiments, the anti-PD-L1 antibody is HLX20. In some embodiments, the anti-PD-L1 antibody is LY3300054.

[0138] In some embodiments, the inhibitor of an immune checkpoint molecule is a small molecule that binds to PD-L1, or a pharma- ceutically acceptable salt thereof. In some embodiments, the inhibitor of an immune checkpoint molecule is a small molecule that binds to and internalizes PD-L1, or a pharma- ceutically acceptable salt thereof. In some embodiments, the inhibitor of an immune checkpoint molecule is a compound selected from those in US2018 / 0179201, US2018 / 0179197, US2018 / 0179179, US2018 / 0179202, US2018 / 0177784, US2018 / 0177870, U.S. Patent Application No. 16 / 369,654 (filed March 29, 2019), and U.S. Patent Application No. 62 / 688,164, each of which is incorporated herein by reference in its entirety, or a pharma- ceutically acceptable salt thereof.

[0139] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of KIR, TIGIT, LAIR1, CD160, 2B4, and TGFRbeta.

[0140] In some embodiments, the inhibitor is MCLA-145.

[0141] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CTLA-4, such as an anti-CTLA-4 antibody. In some embodiments, the anti-CTLA-4 antibody is ipilimumab, tremelimumab, AGEN1884, or CP-675,206.

[0142] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of LAG3, such as an anti-LAG3 antibody. In some embodiments, the anti-LAG3 antibody is BMS-986016, LAG525, INCAGN2385, or eftiragimode alpha (IMP321).

[0143] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is oleclumab.

[0144] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TIGIT. In some embodiments, the inhibitor of TIGIT is OMP-31M32.

[0145] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of VISTA. In some embodiments, the inhibitor of VISTA is JNJ-61610588 or CA-170.

[0146] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of B7-H3. In some embodiments, the inhibitor of B7-H3 is enoblituzumab, MGD009, or 8H9.

[0147] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of KIR. In some embodiments, the inhibitor of KIR is lirilumab or IPH4102.

[0148] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of A2aR. In some embodiments, the inhibitor of A2aR is CPI-444.

[0149] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TGF-beta. In some embodiments, the inhibitor of TGF-beta is travedersen, galcertinib, or M7824.

[0150] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of PI3K-gamma. In some embodiments, the inhibitor of PI3K-gamma is IPI-549.

[0151] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD47. In some embodiments, the inhibitor of CD47 is Hu5F9-G4 or TTI-621.

[0152] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD73. In some embodiments, the inhibitor of CD73 is MEDI9447.

[0153] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD70. In some embodiments, the inhibitor of CD70 is cusatuzumab or BMS-936561.

[0154] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of TIM3, such as an anti-TIM3 antibody. In some embodiments, the anti-TIM3 antibody is INCAGN2390, MBG453, or TSR-022.

[0155] In some embodiments, the inhibitor of an immune checkpoint molecule is an inhibitor of CD20, e.g., an anti-CD20 antibody. In some embodiments, the anti-CD20 antibody is obinutuzumab or rituximab.

[0156] In some embodiments, the agonist of an immune checkpoint molecule is an agonist of OX40, CD27, CD28, GITR, ICOS, CD40, TLR7 / 8, and CD137 (also known as 4-1BB).

[0157] In some embodiments, the agonist of CD137 is urelumab. In some embodiments, the agonist of CD137 is utomirumab.

[0158] In some embodiments, the agonist of the immune checkpoint molecule is an inhibitor of GITR. In some embodiments, the agonist of GITR is TRX518, MK-4166, INCAGN1876, MK-1248, AMG228, BMS-986156, GWN323, MEDI1873, or MEDI6469. In some embodiments, the agonist of the immune checkpoint molecule is an agonist of OX40, such as an OX40 agonist antibody or an OX40L fusion protein. In some embodiments, the anti-OX40 antibody is INCAGN01949, MEDI0562 (tavolimab), MOXR-0916, PF-04518600, GSK3174998, BMS-986178, or 9B12. In some embodiments, the OX40L fusion protein is MEDI6383.

[0159] In some embodiments, the agonist of the immune checkpoint molecule is an agonist of CD40. In some embodiments, the agonist of CD40 is CP-870893, ADC-1013, CDX-1140, SEA-CD40, RO7009789, JNJ-64457107, APX-005M, or ChiLob7 / 4.

[0160] In some embodiments, the agonist of the immune checkpoint molecule is an agonist of ICOS. In some embodiments, the agonist of ICOS is GSK-3359609, JTX-2011, or MEDI-570.

[0161] In some embodiments, the agonist of the immune checkpoint molecule is an agonist of CD28. In some embodiments, the agonist of CD28 is celalizumab.

[0162] In some embodiments, the agonist of the immune checkpoint molecule is an agonist of CD27. In some embodiments, the agonist of CD27 is varlilumab.

[0163] In some embodiments, the agonist of the immune checkpoint molecule is an agonist of TLR7 / 8. In some embodiments, the agonist of TLR7 / 8 is MEDI9197.

[0164] The compounds of the present disclosure can be used in combination with bispecific antibodies. In some embodiments, one of the domains of the bispecific antibody targets PD-1, PD-L1, CTLA-4, GITR, OX40, TIM3, LAG3, CD137, ICOS, CD3, or TGFβ receptor. In some embodiments, the bispecific antibody binds to PD-1 and PD-L1. In some embodiments, the bispecific antibody that binds to PD-1 and PD-L1 is MCLA-136. In some embodiments, the bispecific antibody binds to PD-L1 and CTLA-4. In some embodiments, the bispecific antibody that binds to PD-L1 and CTLA-4 is AK104.

[0165] In some embodiments, the compounds of the present disclosure can be used in combination with one or more metabolic enzyme inhibitors.In some embodiments, the metabolic enzyme inhibitor is an inhibitor of IDO1, TDO, or arginase.Examples of IDO1 inhibitors include epacadostat, NLG919, BMS-986205, PF-06840003, IOM2983, RG-70099, and LY338196.Inhibitors of arginase inhibitors include INCB1158.

[0166] As noted throughout, the additional compounds, inhibitors, drugs, etc. may be combined with the compounds of the invention in a single or continuous dosage form, or may be administered simultaneously or sequentially as separate dosage forms.

[0167] Formulation, Dosage Forms, and Administration When used as a pharmaceutical, the compounds of the present disclosure may be administered in the form of a pharmaceutical composition. Thus, the present disclosure provides a composition comprising a compound of formula I, II, or any of the formulas described herein, a compound listed in any of the claims and described herein, or a pharma- ceutically acceptable salt thereof, or any of these embodiments, and at least one pharma- ceutically acceptable carrier or excipient. These compositions may be prepared in a manner well known to those skilled in the pharmaceutical arts, and may be administered by various routes depending on the area to be treated, depending on whether the indicated treatment is localized or systemic. Administration may be topical (including transdermal, epidermal, ocular, and mucosal delivery, including intranasal, intravaginal, and intrarectal), pulmonary (e.g., by inhalation or insufflation of powder or aerosol, including by nebulizer; intratracheal or intranasal), or oral or parenteral. Parenteral administration includes intravenous, intraarterial, subcutaneous, intraperitoneal, intramuscular injection or infusion; or intracranial, e.g., intrathecal or intraventricular administration. Parenteral administration may be in the form of a single bolus dose or, for example, by a continuous perfusion pump. Pharmaceutical compositions and formulations for topical administration may include transdermal patches, ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

[0168] The present invention also includes pharmaceutical compositions comprising the disclosed compounds or pharma- ceutically acceptable salts thereof as active ingredients in combination with one or more pharma- ceutically acceptable carriers or excipients. In some embodiments, the compositions are suitable for topical administration. In making compositions of the present invention, the active ingredient is typically mixed with an excipient, diluted by an excipient, or enclosed within a carrier, for example, in the form of a capsule, sachet, paper, or other container. When an excipient functions as a diluent, it may be a solid, semi-solid, or liquid material that acts as a vehicle, carrier, or medium for the active ingredient. Thus, the composition may be in the form of tablets, pills, powders, lozenges, sachets, oblates, elixirs, suspensions, emulsions, solutions, syrups, aerosols (as solids or in liquid media), for example, ointments containing up to 10% by weight of the active compound, soft and hard gelatin capsules, suppositories, sterile injectable solutions, and sterile packaged powders.

[0169] In the preparation of the formulation, active compound can be milled to provide suitable particle size, and then combined with other ingredients.If active compound is substantially insoluble, it can be milled to a particle size of less than 200 mesh.If active compound is substantially water-soluble, particle size can be adjusted by milling to provide substantially uniform distribution in the formulation, for example, about 40 mesh.

[0170] The compounds of the invention may be milled using known milling procedures, such as wet milling, to obtain particle sizes suitable for tablet formation and other formulation types. Micronized (nanoparticulate) preparations of the compounds of the invention may be prepared by processes known in the art, see, for example, WO2002 / 000196.

[0171] Some examples of suitable additives include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup and methylcellulose.The formulation may further include lubricants such as talc, magnesium stearate and mineral oil; wetting agents; emulsifying and suspending agents; preservatives such as methyl- and propylhydroxy-benzoates; sweeteners; and flavoring agents.The composition of the present invention may be formulated to provide quick, sustained or delayed release of active ingredient after administration to a patient by using procedures known in the art.

[0172] In some embodiments, the pharmaceutical composition comprises silicified microcrystalline cellulose (SMCC) and at least one compound described herein or a pharma- ceutical acceptable salt thereof. In some embodiments, the silicified microcrystalline cellulose comprises about 98 w / w% microcrystalline cellulose and about 2 w / w% silicon dioxide.

[0173] In some embodiments, the composition is a sustained release composition comprising at least one compound described herein or a pharma- ceutically acceptable salt thereof, and at least one pharma- ceutically acceptable carrier or excipient. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically acceptable salt thereof, and at least one component selected from microcrystalline cellulose, lactose monohydrate, hydroxypropyl methylcellulose, and polyethylene oxide. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and hydroxypropyl methylcellulose. In some embodiments, the composition comprises at least one compound described herein or a pharma- ceutically acceptable salt thereof, and microcrystalline cellulose, lactose monohydrate, and polyethylene oxide. In some embodiments, the composition further comprises magnesium stearate or silicon dioxide. In some embodiments, the microcrystalline cellulose is Avicel PH102™. In some embodiments, the lactose monohydrate is Fast-flo316™. In some embodiments, the hydroxypropyl methylcellulose is hydroxypropyl methylcellulose 2208 K4M (e.g., Methocel K4 M Premier™) and / or hydroxypropyl methylcellulose 2208 K100LV (e.g., Methocel K00LV™). In some embodiments, the polyethylene oxide is polyethylene oxide WSR 1105 (e.g., Polyox WSR 1105™).

[0174] In some embodiments, the composition is produced using a wet granulation process. In some embodiments, the composition is produced using a dry granulation process.

[0175] The compositions may be formulated in unit dosage form, each dosage containing from about 5 to about 1,000 mg (1 g), more usually from about 100 mg to about 500 mg, of the active ingredient. In some embodiments, each dosage contains about 10 mg of the active ingredient. In some embodiments, each dosage contains about 50 mg of the active ingredient. In some embodiments, each dosage contains about 25 mg of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect in association with suitable pharmaceutical excipients.

[0176] The components used to formulate the pharmaceutical composition are of high purity and substantially free of potentially harmful contaminants (e.g., at least National Food grade, generally at least analytical grade, more typically at least pharmaceutical grade). Particularly for human consumption, the composition is preferably manufactured or formulated under Good Manufacturing Practice regulations as defined in applicable regulations of the U.S. Food and Drug Administration. For example, suitable formulations may be sterile and / or substantially isotonic, and / or in full compliance with all Good Manufacturing Practice regulations of the U.S. Food and Drug Administration.

[0177] The active compounds may be effective over a wide dosage range and are generally administered in a therapeutically effective amount. However, it will be understood that the amount of compound actually administered will usually be determined by the physician according to the appropriate circumstances, including the condition to be treated, the route of administration selected, the actual compound administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, and the like.

[0178] The therapeutic dosage of the compounds of the present invention may vary, for example, according to the particular use for which the treatment is made, the manner of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compounds of the present invention in a pharmaceutical composition may vary depending on a number of factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention may be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg of body weight per day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg of body weight per day. The dosage is likely to depend on variables, such as the type and extent of progression of the disease or disorder, the overall health of the particular patient, the relative biological availability of the selected compound, the formulation of excipients, and its route of administration. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0179] For the preparation of solid compositions, such as tablets, the main active ingredient is mixed with pharmaceutical excipients to form a solid preformulation composition containing a homogeneous mixture of the compound of the present invention.When these preformulation compositions are referred to as homogeneous, the active ingredient is typically uniformly distributed throughout the composition, so that the composition can be easily and uniformly subdivided into effective unit dosage forms, such as tablets, pills, and capsules.The solid preformulation is then subdivided into unit dosage forms of the above-mentioned type, such as about 0.1 to about 1000 mg of the active ingredient of the present invention.

[0180] The tablets or pills of the present invention may be coated or otherwise compounded to provide a dosage form that provides the advantage of prolonged action.For example, the tablet or pill may comprise an inner dosage and an outer dosage component, the latter being in the form of a coating over the former.The two components may be separated by an enteric layer that serves to resist disintegration in the stomach and allows the inner component to pass intact into the duodenum or be released in a delayed manner.A variety of materials may be used for such enteric layers or coatings, including a number of polymeric acids and mixtures of polymeric acids with materials such as shellac, cetyl alcohol and cellulose acetate.

[0181] Liquid forms into which the compounds and compositions of the present invention may be incorporated for administration orally or by injection include aqueous solutions, suitably flavored syrups, aqueous or oily suspensions, and emulsions flavored with edible oils, such as cottonseed oil, sesame oil, coconut oil, or peanut oil, as well as elixirs and similar pharmaceutical vehicles.

[0182] Compositions for inhalation or insufflation include solutions and suspensions in pharma- ceutically acceptable, aqueous or organic solvents or mixtures thereof, as well as powders. Liquid or solid compositions may contain suitable pharma- ceutically acceptable additives as described above. In some embodiments, the compositions are administered by oral or nasal respiratory routes for localized or systemic effect. Compositions may be nebulized by using inert gases. Nebulized solutions may be breathed directly from the nebulizing device, or the nebulizing device may be attached to a face mask, tent, or intermittent positive pressure breathing machine. Solution, suspension, or powder compositions may be administered orally or nasally from a device that delivers the formulation in an appropriate manner.

[0183] Topical formulations may include one or more conventional carriers. In some embodiments, ointments may include water and one or more hydrophobic carriers selected from, for example, liquid paraffin, polyoxyethylene alkyl ether, propylene glycol, white petrolatum, and the like. Cream carrier compositions may be based on water in combination with glycerol and one or more other components, for example, glycerol monostearate, PEG-glycerol monostearate, and cetylstearyl alcohol. Gels may be formulated using isopropyl alcohol and water in suitable combination with other components, for example, glycerol, hydroxyethylcellulose, and the like. In some embodiments, topical formulations include at least about 0.1, at least about 0.25, at least about 0.5, at least about 1, at least about 2, or at least about 5 wt% of the compound of the present invention. Topical formulations may be suitably packaged, for example, in 100 g tubes, optionally associated with instructions for use to treat a selected indication, for example, psoriasis or other skin conditions.

[0184] The amount of compound or composition administered to a patient will vary depending on what is being administered, the purpose of the administration, e.g., prophylaxis or therapy, the condition of the patient, the mode of administration, etc. In therapeutic applications, the composition may be administered to a patient already suffering from a disease in an amount sufficient to cure or at least partially arrest the symptoms of the disease and its complications. The effective dose will depend on the disease state being treated, and will be at the discretion of the attending clinician depending on factors such as the severity of the disease, the age, weight, and general condition of the patient, etc.

[0185] The compositions administered to a patient may be in the form of pharmaceutical compositions described above. These compositions may be sterilized by conventional aseptic techniques or sterile filtered. Aqueous solutions may be packaged or lyophilized for use as is, and lyophilized preparations are combined with a sterile aqueous carrier before administration. The pH of the compound preparations will typically be between 3 and 11, more preferably between 5 and 9, and most preferably between 7 and 8. It will be understood that certain use of the above-mentioned additives, carriers, or stabilizers will result in the formation of pharmaceutical salts.

[0186] The therapeutic dosage of the compounds of the present invention may vary, for example, according to the particular use for which the treatment is made, the manner of administration of the compound, the health and condition of the patient, and the judgment of the prescribing physician. The proportion or concentration of the compounds of the present invention in a pharmaceutical composition may vary depending on a number of factors, including dosage, chemical characteristics (e.g., hydrophobicity), and route of administration. For example, the compounds of the present invention may be provided in an aqueous physiological buffer solution containing about 0.1 to about 10% w / v of the compound for parenteral administration. Some typical dosage ranges are about 1 μg / kg to about 1 g / kg of body weight per day. In some embodiments, the dosage range is about 0.01 mg / kg to about 100 mg / kg of body weight per day. The dosage is likely to depend on variables, such as the type and extent of progression of the disease or disorder, the overall health of the particular patient, the relative biological availability of the selected compound, the formulation of excipients, and its route of administration. Effective doses may be extrapolated from dose-response curves derived from in vitro or animal model test systems.

[0187] Labeled Compounds and Assay Methods Another aspect of the present invention relates to the labeled (radiolabeled, fluorescently labeled, etc.) compounds of the present disclosure that will be useful in both in vitro and in vivo imaging techniques as well as assays for localizing and quantifying KRAS protein in tissue samples, including human, and for identifying KRAS ligands by inhibiting the binding of the labeled compounds. Substitution of one or more atoms of the compounds of the present disclosure may also be useful in generating special ADME (adsorption, distribution, metabolism, and excretion). Thus, the present invention includes KRAS binding assays that include such labeled or substituted compounds.

[0188] The present disclosure further includes isotopically labeled compounds of the present disclosure. An "isotopically" or "radiolabeled" compound is a compound of the present disclosure in which one or more atoms have been replaced or substituted by an atom having an atomic mass or mass number different from the atomic mass or mass number typically found in nature (i.e., naturally occurring). Suitable radionuclides that may be incorporated into the disclosed compounds include, but are not limited to, 2 H (also written as D for deuterium), 3 H (also written as T for tritium), 11 C. 13 C. 14 C. 13 N, 15 N, 15 O. 17 O. 18 O. 18 F, 35 S, 36 Cl, 82 Br, 75 Br, 76 Br, 77 Br, 123 I, 124 I, 125 I and 131 For example, one or more hydrogen atoms in the compounds of the present disclosure may be replaced by a deuterium atom (e.g., C of formula I, II, or any formula provided herein). 1~6One or more hydrogen atoms of an alkyl group may be optionally replaced with a deuterium atom, e.g., -CD3 is replaced with -CH3.) In some embodiments, an alkyl group of Formula I, II, or any formula provided herein may be perdeuterated.

[0189] One or more constituent atoms of the compounds presented herein may be replaced or substituted with an atomic isotope in natural or non-natural abundance ratio. In some embodiments, the compounds contain at least one deuterium atom. In some embodiments, the compounds contain two or more deuterium atoms. In some embodiments, the compounds contain 1-2, 1-3, 1-4, 1-5, or 1-6 deuterium atoms. In some embodiments, all of the hydrogen atoms in the compounds may be replaced or substituted with deuterium atoms.

[0190] Synthetic methods for incorporating isotopes into organic compounds are known in the art (Deuterium Labeling in Organic Chemistry by Alan F. Thomas (New York, NY, Appleton-Century-Crofts, 1971; The Renaissance of H / D Exchange by Jens Atzrodt, Volker Derdau, Thorsten Fey and Jochen Zimmermann, Angew. Chem. Int. Ed. 2007, 7744-7765; The Organic Chemistry of Isotopic Labelling by James R. Hanson, Royal Society of Chemistry, 2011). Isotopically labeled compounds may be used in a variety of studies, e.g., NMR spectroscopy, metabolic experiments, and / or assays.

[0191] Substitution with heavier isotopes, such as deuterium, may provide certain therapeutic advantages due to superior metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and may therefore be preferred in some circumstances. (See, for example, A. Kerekes et.al. J. Med. Chem. 2011, 54, 201-210; R. Xu et.al. J. Label Compd. Radiopharm. 2015, 58, 308-312). In particular, substitution at one or more metabolic sites may provide one or more of the therapeutic advantages.

[0192] The radionuclide incorporated in the present radiolabeled compounds will depend on the particular application of that radiolabeled compound. For example, for in vitro adenosine receptor labeling and competition assays: 3 H, 14 C. 82 Br, 125 I, 131 I or 35 Compounds incorporating S may be useful. For radioimaging applications, 11 C. 18 F, 125 I, 123 I, 124 I, 131 I, 75 Br, 76 Br or 77 Br may be useful.

[0193] A "radiolabel" or "labeled compound" is understood to be a compound into which at least one radionuclide has been incorporated. In some embodiments, the radionuclide is 3 H, 14 C. 125 I, 35 S and 82 Br is selected.

[0194] The present disclosure may further include synthetic methods for incorporating radioisotopes into the compounds of the present disclosure. Synthetic methods for incorporating radioisotopes into organic compounds are well known in the art, and one of ordinary skill in the art would readily recognize methods applicable to the compounds of the present disclosure.

[0195] The labeled compounds of the present invention may be used in screening assays to identify and / or evaluate compounds. For example, a newly synthesized or identified labeled compound (i.e., test compound) can be evaluated for its ability to bind to KRAS protein by monitoring its concentration change when it is contacted with KRAS via tracking of the label. For example, a test compound (labeled) can be evaluated for its ability to reduce the binding of another compound (i.e., standard compound) that is known to bind to KRAS protein. Thus, the ability of the test compound to compete with the standard compound for direct binding to KRAS protein correlates with its binding affinity. Conversely, in some other screening assays, the standard compound is labeled and the test compound is not labeled. Thus, the concentration of the labeled standard compound is monitored to evaluate the competition between the standard compound and the test compound, thereby ascertaining the relative binding affinity of the test compound.

[0196] kit The present disclosure also includes pharmaceutical kits useful in treating or preventing diseases or disorders associated with the activity of KRAS, such as cancer or infectious diseases, which include one or more containers containing a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I, II, or any of these embodiments. Such kits may further include one or more of various conventional pharmaceutical kit components, such as a container containing one or more pharma- ceutically acceptable carriers, additional containers, etc., as would be readily apparent to one skilled in the art. Instructions, either as an insert or label indicating the amount of components to be administered, guidelines for administration, and / or guidelines for mixing components may also be included in the kit.

[0197] The present invention will be described in more detail by specific examples.The following examples are provided for illustrative purposes and are not intended to limit the invention in any way.Those skilled in the art will easily recognize various non-critical parameters that can be changed or modified to obtain substantially the same results.The compound of the example is found to inhibit the activity of KRAS according to at least one assay described herein. EXAMPLES

[0198] The experimental procedures for the compounds of the present invention are provided below. Preparative LC-MS purification of some of the compounds prepared was carried out on a Waters mass-directed fractionation system. The basic instrument settings, protocols, and control software for the operation of these systems are described in detail in the literature. See, e.g., "Two-Pump At Column Dilution Configuration for Preparative LC-MS", K. Blom, J. Combi. Chem., 4, 295 (2002); "Optimizing Preparative LC-MS Configurations and Methods for Parallel Synthesis Purification", K. Blom, R. Sparks, J. Doughty, G. Everlof, T. Haque, A. Combs, J. Combi. Chem., 5, 670 (2003); and "Preparative LC-MS Purification: Improved Compound Specific Method Optimization", K. Blom, B. Glass, R. Sparks, A. Combs, J. Combi. Chem., 6, 874-883 (2004). Separated compounds are typically subjected to analytical liquid chromatography mass spectrometry (LCMS) to confirm purity.

[0199] The isolated compounds were typically subjected to analytical liquid chromatography mass spectrometry (LCMS) to confirm purity under the following conditions: Instrument: Agilent 1100 series, LC / MSD; Column: Waters Sunfire™ C 18 Particle size 5 μm, 2.1×5.0 mm, buffer: mobile phase A: 0.025% TFA in water and mobile phase B: acetonitrile; gradient 2% to 80% of B in 3 min at a flow rate of 2.0 mL / min.

[0200] Some of the prepared compounds were also separated on a preparative scale by reversed-phase high performance liquid chromatography (RP-HPLC) with MS detection or flash chromatography (silica gel) as shown in the examples. Typical preparative reversed-phase high performance liquid chromatography (RP-HPLC) column conditions are as follows:

[0201] pH=2 purification: Waters Sunfire(TM) C 18 Particle size 5 μm, 19×100 mm column, eluted with mobile phase A: 0.1% TFA (trifluoroacetic acid) in water and mobile phase B: acetonitrile; flow rate was 30 mL / min, separation gradient was optimized for each compound using compound specific method optimization protocol as described in literature [see “Preparative LCMS Purification: Improved Compound Specific Method Optimization”, K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)]. Typically, the flow rate used for the 30×100 mm column was 60 mL / min.

[0202] pH=10 Purification: Waters XBridge C 18Particle size 5 μm, 19×100 mm column, eluted with mobile phase A: 0.15% NH4OH in water and mobile phase B: acetonitrile; flow rate was 30 mL / min, separation gradient was optimized for each compound using compound specific method optimization protocol as described in literature [see “Preparative LCMS Purification: Improved Compound Specific Method Optimization”, K. Blom, B. Glass, R. Sparks, A. Combs, J. Comb. Chem., 6, 874-883 (2004)]. Typically, the flow rate used for the 30×100 mm column was 60 mL / min.

[0203] The following abbreviations may be used herein: AcOH (acetic acid); AcO (acetic anhydride); aq. (aqueous solution); atm. (atmosphere); Boc (t-butoxycarbonyl); BOP ((benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate); br (broad); Cbz (carboxybenzyl); calc. (calculated); d (doublet); dd (doublet of doublets); DBU (1,8-diazabicyclo[5.4.0]undec-7-ene); DCM (dichloromethane); DIAD (N,N'-diisopropyl azidodicarboxylate); DIEA (N,N-diisopropylethylamine); DIPEA (N,N-diisopropylethylamine); DIBAL (diisobutylaluminum hydride); DMF (N,N-dimethylformamide); Et (ethyl); EtOAc (ethyl acetate); FCC (flash column chromatography); g (grams); h (hours); HATU (N,N ,N',N'-Tetramethyl-O-(7-azabenzotriazol-1-yl)uronium hexafluorophosphate;HCl (hydrochloric acid);HPLC (high performance liquid chromatography);Hz (hertz);J (coupling constant);LCMS (liquid chromatography-mass spectrometry);LDA (lithium diisopropylamide);m (multiplet);M (molar);mCPBA (3-chloroperoxybenzoic acid);MS (mass spectrometry);Me (methyl);MeCN (acetonitrile);MeOH (methanol);mg (milligram);min. (minute);mL (milliliter);mmol (millimolar);N (normal);NCS (N-chlorosuccinimide);NEt3 (triethylamine);nM (nanomolar);NMP (N-methylpyrrolidone);NMR (nuclear magnetic resonance spectroscopy);OTf (trifluoromethanesulfonate);Ph (phenyl);pM (picomolar);PPT (precipitate);RP-HPLC (reversed-phase high performance liquid chromatography);rt(room temperature), s (singlet); t (triplet or tertiary); TBS (tert-butyldimethylsilyl); tert (tertiary); tt (triplet of triplets); TFA (trifluoroacetic acid); THF (tetrahydrofuran); μg (microgram); μL (microliter); μM (micromolar); wt% (weight percent). Brine is a saturated aqueous solution of sodium chloride. in vacuo is under vacuum.

[0204] Intermediate 1. 7-Bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline [ka]

[0205] Step 1. 2-Amino-4-bromo-3-fluoro-5-iodobenzoic acid [ka] To a solution of 2-amino-4-bromo-3-fluorobenzoic acid (10.0 g, 42.7 mmol) in DMF (100 mL) was added NIS (9.61 g, 42.7 mmol) and the reaction was stirred at 80° C. for 6 h. The mixture was cooled in ice water, water (150 mL) was added, stirred for 20 min, and the precipitate was filtered, washed with water, and dried to give the desired product as a solid. C7H5BrFINO2 (M+H) + LCMS calculated for: m / z = 359.9; found 359.8.

[0206] Step 2. 7-Bromo-8-fluoro-6-iodo-2H-benzo[d][1,3]oxazine-2,4(1H)-dione [ka] To a solution of 2-amino-4-bromo-3-fluoro-5-iodobenzoic acid (8.4 g, 23.34 mmol) in 1,4-dioxane (200 mL) was added triphosgene (6.34 g, 21.37 mmol) and stirred at 100° C. for 1 h. After cooling to room temperature, ice was added until a solid precipitated. The mixture was then thoroughly diluted with water (final volume about 400 mL) and the solid was collected by filtration and then air-dried. The crude product was used in the next step without further purification.

[0207] Step 3. 7-Bromo-8-fluoro-6-iodo-3-nitroquinoline-2,4-diol [ka] To a solution of ethyl 2-nitroacetate (4.62 g, 17.36 mmol) in toluene (10.0 mL) at room temperature, DIPEA (6.06 ml, 34.7 mmol) was added and stirred for 10 min. Then, 7-bromo-8-fluoro-6-iodo-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (6.7 g, 17.36 mmol) was added to the reaction mixture and the reaction was stirred at 95° C. for 3 h. The reaction was cooled with ice water, after which 1N HCl (40 mL) was added. The solid precipitate was collected via filtration and then washed with a small amount of ethyl acetate to give the desired product as a yellow solid (6 g, 81%). C9H4BrFIN2O4 (M+H) + LCMS calculated for: m / z = 428.8; found 428.8.

[0208] Step 4. 7-Bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline [ka] To a mixture of 7-bromo-8-fluoro-6-iodo-3-nitroquinoline-2,4-diol (4.51 g, 10.51 mmol) in POCl3 (4.9 mL, 52.6 mmol) was added DIPEA (3.67 mL, 21.03 mmol) and the reaction was stirred at 105° C. for 3 h. The solvent was removed under reduced pressure and then azeotroped with toluene three times to give the crude material, which was used in the next step without further purification.

[0209] Intermediate 2. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-iodo-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka]

[0210] Step 1. tert-Butyl (2S,4S)-4-((3-amino-7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-8-fluoro-6-iodoquinolin-4-yl)amino)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] To a solution of 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline (Intermediate 1) (20.49 g, 44 mmol) and tert-butyl (2S,4S)-4-amino-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate (Intermediate 6) (13.83 g, 44 mmol) in CHCl (100 mL) at room temperature was added DIPEA (15.37 ml, 88 mmol). The reaction was stirred at 50° C. for 3 h. Upon complete conversion, N,N,3-trimethylazetidin-3-amine dihydrochloride (9.98 g, 52.8 mmol) and an additional 2 equivalents of DIPEA (15.37 mL, 88 mmol) were added to the reaction mixture and stirred at 50° C. overnight. The reaction contents were transferred to a separatory funnel and washed with saturated NH4Cl (200 mL) and brine (100 mL). The organic phase was dried over MgSO4 and concentrated.

[0211] The concentrated residue was redissolved in MeOH (50 mL), CHCl (10 mL), and aqueous ammonium hydroxide (57 mL, 440 mmol). Sodium dithionite (23 g, 132 mmol) was added in one portion and the reaction was stirred vigorously at room temperature overnight. Upon completion, the reaction was quenched by the addition of HO (100 mL) and extracted with CHCl (100 mL). The organic phase was washed twice with HO, dried over NaSO, and then concentrated to give the desired diamine product as a red viscous oil (18.21 g, 56% for two steps). C 31 H 46 BrFIN6O4(M+H) + LCMS calculated for: m / z = 791.2; found: 791.1.

[0212] Step 2. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-iodo-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] The crude diamine from step 1 (18.21 g, 23 mmol) was dissolved in glacial acetic acid (57.9 mL, 1012 mmol). Sodium nitrite (2.38 g, 34.5 mmol) was added in one portion. The reaction was stirred at room temperature for 2 h. Upon completion, the reaction contents were poured into vigorously stirred ice water. The precipitated solid was collected via filtration and washed with NaHCO3, water, and diethyl ether. The solid was then dried under reduced pressure to give the desired product as a brown solid (15 g, 81% yield). 31 H 43 BrFIN7O4(M+H) + LCMS calculated for: m / z = 802.2; found 802.1.

[0213] Intermediate 3. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka]

[0214] Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] Intermediate 2 (1.10 g, 1.37 mmol), copper(I) iodide (39 mg, 0.21 mmol), 1,10-phenanthroline (37 mg, 0.21 mmol), and potassium fluoride (239 mg, 411 mmol) were placed in a pressure vessel. DMSO (2.74 mL) was added. The vessel was flushed with N2 for 5 min. Trimethylborate (0.46 mL, 4.11 mmol) and trimethyl(trifluoromethyl)silane (0.61 mL, 4.11 mmol) were added, after which the pressure vessel was sealed and heated to 80 °C overnight. The vessel was cooled to room temperature, then cooled in an ice bath and carefully opened. The reaction mixture was diluted with EtOAc (20 mL), washed with NaHCO3, brine, and concentrated. The crude product was purified on silica gel (20 g, 50-100% EtOAc in CH2Cl2) to give a brown solid (634 mg, 62% yield). 32 H 43 BrF4N7O4(M+H) + LCMS calculated for: m / z = 744.3; found: 744.2.

[0215] Step 2. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate (6.5 g, 8.7 mmol) was dissolved in dioxane (20 mL) and lithium hydroxide solution (7.27 mL, 6M in H2O) was added. The reaction was heated to 80° C. overnight. Upon complete hydrolysis of the tert-butyl ester, saturated NH4Cl (100 mL) was added and the reaction was extracted with EtOAc (3×100 mL). The combined organic layers were dried over Na2SO4 and concentrated to dryness to give 2-((2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(tert-butoxycarbonyl)piperidin-2-yl)acetic acid. 28 H 35 BrF4N7O4(M+H) + LCMS calculated for: m / z = 688.2; found 688.1.

[0216] The carboxylic acid was redissolved in THF (20 mL). DIPEA (3.1 mL, 17.5 mmol) was added and the reaction mixture was cooled to 0° C. Then, isobutyl chloroformate (1.7 mL, 13.1 mmol) was added. After stirring at 0° C. for 20 min, ammonium hydroxide (11.3 mL, 87 mmol) was added and the mixture was stirred for an additional 10 min. Upon completion, the reaction was diluted with EtOAc (20 mL), washed with brine, dried over MgSO4, and concentrated to give tert-butyl (2S,4S)-2-(2-amino-2-oxoethyl)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 28 H 36 BrF4N8O3(M+H)+ LCMS calculated for: m / z = 687.2; found 687.4.

[0217] The crude amide was redissolved in THF (20 mL) and cooled to 0 °C. Triethylamine (4.9 mL, 34.9 mmol) and TFAA (1.8 mL, 13.1 mol) were added successively. After stirring for 1 h, the reaction was quenched by addition of aqueous NaHCO3 (50 mL), extracted with EtOAc (50 mL), dried over Na2SO4, concentrated, and purified on silica (100 g, 0-100% EtOAc in CH2Cl2) to give the title product as a yellow solid (4.3 g, 74% yield over three steps). 28 H 34 BrF4N8O2(M+H) + LCMS calculated for: m / z = 669.2; found 669.4.

[0218] Intermediate 4. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka]

[0219] Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] To a solution of 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline (Intermediate 1) (2.56 g, 5.5 mmol) and tert-butyl (2S,4S)-4-amino-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate (Intermediate 6) (1.73 g, 5.5 mmol) in CHCl (10 mL) at room temperature was added DIPEA (1.92 ml, 11 mmol). The reaction was stirred at 50° C. for 3 h. Upon complete conversion, the reaction contents were transferred to a separatory funnel and washed with saturated NHCl (50 mL) and brine (50 mL). The organic phase was dried over MgSO and concentrated.

[0220] The concentrated residue was redissolved in MeOH (5 mL), CHCl (5 mL), and aqueous ammonium hydroxide (7.3 mL, 55 mmol). Sodium dithionite (2.88 g, 16.5 mmol) was added in one portion and the reaction was stirred vigorously at room temperature overnight. Upon completion, the reaction was quenched by the addition of HO (10 mL) and extracted with CHCl (50 mL). The organic phase was washed twice with HO, dried over NaSO, and then concentrated to give the desired diamine product.

[0221] The crude diamine was dissolved in glacial acetic acid (7.0 mL). Sodium nitrite (0.76 g, 11 mmol) was added in one portion. The reaction was stirred at room temperature for 20 min. Upon completion, the reaction contents were poured into vigorously stirred ice water. The reaction mixture was extracted with DCM. The crude product was purified on silica gel (50 g, 0-100% EtOAc in CH2Cl2) to give the desired product (1.8 g, 70% yield). C 25 H 30 BrClFIN5O4(M+H) + LCMS calculated for: m / z = 724.0; found: 724.0.

[0222] Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] tert-Butyl (2S,4S)-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate (1.44 g, 1.99 mmol) was dissolved in CHCl (5 mL) and MeOH (5 mL) and stirred at room temperature until homogeneous. Sodium thiomethoxide (0.28 g, 3.97 mmol) was added in one portion. After stirring for 1 h, the reaction was quenched with saturated NHCl (10 mL) and extracted with EtOAc (20 mL) followed by washing with NaHCO. The combined organic layers were dried over MgSO, filtered and concentrated. C 26 H 33 BrFIN5O4S (M+H) + LCMS calculated for: m / z = 736.1; found: 736.0.

[0223] Intermediate 5. tert-Butyl 4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Intermediate 4, using tert-butyl 4-aminopiperidine-1-carboxylate instead of tert-butyl (2S,4S)-4-amino-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate. 20 H 23 BrFIN5O2S (M+H) +LCMS calculated for: m / z = 622.0; found: 622.1.

[0224] Intermediate 6. tert-Butyl (2S,4S)-4-amino-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka]

[0225] Step 1. (R)-tert-Butyl 6-cyano-5-hydroxy-3-oxohexanoate [ka] A solution of 2.0M LDA (100 mL, 200 mmol) in anhydrous THF (223 mL) was cooled to -78°C for 1 hour, then tert-butyl acetate (26.9 mL, 200 mmol) was added dropwise with stirring over 20 minutes. After maintaining at -78°C for an additional 40 minutes, a solution of ethyl (R)-4-cyano-3-hydroxybutyrate (10.5 g, 66.8 mmol) was added dropwise. The mixture was allowed to stir at -40°C for 4 hours, after which an appropriate amount of HCl (2M) was added to the mixture, keeping the pH at about 6. The temperature of the mixture was maintained at -10°C during this quench. Upon completion, the temperature of the mixture was cooled to 0°C. The mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic layers were washed with NaHCO3 (100 mL) and brine (100 mL), dried over anhydrous Na2SO4, and concentrated to give the material as a yellow oil (15.0 g, 99% yield).

[0226] Step 2. (2S,4R)-tert-butyl 2-(2-(tert-butoxy)-2-oxoethyl)-4-hydroxypiperidine-1-carboxylate [ka] A solution of (R)-tert-butyl 6-cyano-5-hydroxy-3-oxohexanoate (15.0 g, 66.0 mmol) in acetic acid (110 ml) was treated with platinum(IV) oxide hydrate (0.868 g, 3.30 mmol). The Parr bottle was evacuated and filled with H2 three times and stirred under H2 atmosphere (45 psi, filled four times) at 22 °C for 3 h. The mixture was filtered through Celite and the filter cake was washed with EtOH. The filtrate was concentrated to give the product in a cis:trans diastereomeric ratio of approximately 9:1.

[0227] The residue was dissolved in methanol (100 mL), followed by the addition of Boc anhydride (15.32 ml, 66.0 mmol) and sodium carbonate (13.99 g, 132 mmol). The reaction mixture was stirred at room temperature overnight. The mixture was filtered and concentrated. The residue was purified on a silica gel column to give the desired product (11.7 g, 56%). 16 H 29 NNaO5(M+Na) + LCMS calculated for: m / z=338.2; found 338.2.

[0228] Step 3. (2S,4S)-tert-Butyl 4-azido-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] To a solution of tert-butyl (2S,4R)-2-(2-(tert-butoxy)-2-oxoethyl)-4-hydroxypiperidine-1-carboxylate (2.10 g, 6.66 mmol) in DCM (33 ml) at 0° C., TEA (1.58 ml, 11.32 mmol) and Ms-Cl (0.67 mL, 8.66 mmol) were added. After stirring for 1 h, the reaction was diluted with water, the organic layer was separated, dried over Na2SO4, filtered and concentrated. The resulting residue was dissolved in DMF, sodium azide (1.3 g, 20 mmol) was added and the reaction mixture was heated at 70° C. for 5 h. After cooling to room temperature, the reaction was diluted with EtOAc and water. The organic layer was separated, dried over Na2SO4, filtered and concentrated. The residue was purified on a silica gel column to give the desired product (1.90 g, 84%). (Product Boc)C 11 H 21 N4O2(M+H) + LCMS calculated for: m / z=241.2; found 241.2.

[0229] Step 4. (2S,4S)-tert-Butyl 4-amino-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] To a solution of tert-butyl (2S,4S)-4-azido-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate (1.9 g, 5.58 mmol) in methanol (27.9 ml) was added 10% palladium on carbon (0.594 g, 0.558 mmol). The reaction mixture was evacuated under vacuum, charged with H2, and stirred at room temperature for 2 h. The reaction mixture was filtered through a pad of Celite and washed with methanol. The filtrate was concentrated and used as is (1.5 g, 85%). C 16 H 31 N2O4(M+H) + LCMS calculated for: m / z = 315.2; found: 315.2.

[0230] Intermediate 7. tert-Butyl (2S,4S)-4-amino-2-(cyanomethyl)piperidine-1-carboxylate [ka]

[0231] Step 1. tert-Butyl (2S,4S)-4-(((benzyloxy)carbonyl)amino)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] To a stirred solution of intermediate 6 (5.2 g, 16.54 mmol) in CH2Cl2 (80.0 mL) was added N-(benzyloxy-carbonyloxy)succinimide (4.95 g, 19.85 mmol) followed by DIPEA (4.33 mL, 24.81 mmol). The mixture was stirred at room temperature for 2 h and then diluted with water. The mixture was extracted with water and brine. The combined organic layers were dried over MgSO4, filtered, concentrated and used directly in the next step without further purification.

[0232] Step 2. 2-((2S,4S)-4-(((benzyloxy)carbonyl)amino)piperidin-2-yl)acetic acid [ka] The concentrated residue from step 1 was taken up in CH2Cl2 (80.0 mL) and TFA (50.0 mL). The mixture was stirred at room temperature overnight, then concentrated to dryness and used directly in the next step without further purification.

[0233] Step 3. 2-((2S,4S)-4-(((benzyloxy)carbonyl)amino)-1-(tert-butoxycarbonyl)piperidin-2-yl)acetic acid [ka] The concentrated residue from step 2 was taken up in CHCl (80.0 mL) and triethylamine (23.1 mL, 165 mmol) was added slowly. The mixture was stirred at room temperature for 5 min, then Boc anhydride (4.33 g, 19.85 mmol) was added. The mixture was stirred at room temperature for an additional 30 min. Additional Boc anhydride was added if necessary. Upon complete conversion, the mixture was acidified to pH 4-5 and then extracted with EtOAc. The combined organic layers were dried over MgSO, filtered, concentrated, and used directly in the next step without further purification.

[0234] Step 4. tert-Butyl (2S,4S)-2-(2-amino-2-oxoethyl)-4-(((benzyloxy)carbonyl)amino)piperidine-1-carboxylate [ka] The concentrated residue from step 3 was taken up in THF (80.0 mL) and DIPEA (8.67 mL, 49.6 mmol). After the mixture was cooled to 0° C., isobutyl chloroformate (5.43 mL, 41.3 mmol) was added slowly. After the mixture was stirred at 0° C. for an additional 20 min, ammonium hydroxide (28% in water, 23.0 mL, 165 mmol) was added to the mixture. After stirring for an additional 5 min, the mixture was diluted with brine and extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered, concentrated, and purified by column chromatography (0-8% MeOH in DCM).

[0235] Step 5. tert-Butyl (2S,4S)-4-(((benzyloxy)carbonyl)amino)-2-(cyanomethyl)piperidine-1-carboxylate [ka] The purified product from step 4 was taken up in THF (80.0 mL) and triethylamine (6.0 mL, 43 mmol). After the mixture was cooled to 0° C., TFAA (3.5 mL, 24.8 mmol) was added slowly. The mixture was stirred at 0° C. for an additional 30 min, then diluted with EtOAc and extracted with brine. The combined organic layers were dried over MgSO4, filtered, concentrated to dryness, and purified on silica gel to give the desired product (5.12 g, 83% for 5 steps). 16 H 20 N3O4(M+H-tert-butyl) + LCMS calculated for: m / z = 318.1; found: 318.1.

[0236] Step 6. tert-Butyl (2S,4S)-4-amino-2-(cyanomethyl)piperidine-1-carboxylate [ka] A round-bottom flask equipped with a stir bar was charged with tert-butyl (2S,4S)-4-(((benzyloxy)carbonyl)amino)-2-(cyanomethyl)piperidine-1-carboxylate (5.12 g, 13.71 mmol), palladium on carbon (10 wt%, 2.92 g, 2.74 mmol), and MeOH (45 mL). The round-bottom flask was evacuated and backfilled with H (this process was repeated a total of three times), a balloon of H was attached, and the mixture was stirred vigorously at room temperature for 1.5 h. The mixture was then filtered over Celite, and the solids were washed with EtOAc. The filtrate was concentrated and used directly in the next step without further purification.

[0237] Intermediate 8. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka]

[0238] Step 1. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] To a solution of 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline (Intermediate 1) (10 g, 21.5 mmol) and tert-butyl (2S,4S)-4-amino-2-(cyanomethyl)piperidine-1-carboxylate (Intermediate 7) (5.15 g, 21.5 mmol) in MeCN (100 mL) at room temperature was added DIPEA (5.62 ml, 32 mmol). The reaction was stirred at 50° C. for 3 hours. Upon complete conversion, the reaction mixture was concentrated.

[0239] The residue was dissolved in MeOH (80 mL) and sodium thiomethoxide (3.0 g, 43 mmol) was added in one portion at room temperature. Upon complete conversion, the reaction mixture was diluted with water, filtered and washed with water to give a brown solid.

[0240] The solid was redissolved in MeOH (80 mL), CHCl (20 mL), and aqueous ammonium hydroxide (28 mL, 440 mmol). Sodium dithionite (11.2 g, 132 mmol) was then added in one portion and the reaction was stirred vigorously at room temperature. Upon completion, the reaction was quenched by the addition of H0 (100 mL) and extracted with CHCl (100 mL). The organic phase was washed twice with H0, dried over NaSO, and then concentrated to give the desired diamine product as a red viscous oil.

[0241] The crude diamine was dissolved in glacial acetic acid (50 mL, 875 mmol). Sodium nitrite (2.97 g, 43.0 mmol) was added in one portion. The reaction was stirred at room temperature for 2 h. Upon completion, the reaction contents were poured into vigorously stirred ice water. The precipitated solid was collected via filtration and washed with NaHCO3, water, and diethyl ether. The solid was then dried under reduced pressure to give the desired product as a brown solid (7.1 g, 50% yield). 22 H 24 BrFIN6O2S (M+H) + LCMS calculated for: m / z = 661.0; found: 660.9.

[0242] Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] A screw-capped vial equipped with a magnetic stir bar was charged with tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (1.19 g, 1.832 mmol), methylboronic acid (1.096 g, 18.32 mmol), tripotassium phosphate (1.166 g, 5.49 mmol), and bis(triphenylphosphine)-palladium(II) chloride (257 mg, 0.366 mmol), followed by dioxane (10.0 mL) and water (2.0 mL). The vial was sealed with a Teflon-lined septum and evacuated and backfilled with nitrogen, a process repeated a total of three times. The reaction was then stirred at 90° C. for 3 h. After cooling to room temperature, the mixture was diluted with brine and extracted with EtOAc. The combined organic layers were dried over MgSO4, filtered, concentrated to dryness, and purified on silica gel to give the desired product. 23 H 27 BrFN6O2S (M+H)+ LCMS calculated for: m / z = 549.1; found: 549.1.

[0243] Example 1a and Example 1b. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0244] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] Intermediate 3 (1.3 g, 1.9 mmol), XPhos Pd G2 (76 mg, 0.097 mmol), (5-fluoroquinolin-8-yl)boronic acid (408 mg, 2.1 mmol), K3PO4 (1.24 g, 5.83 mmol), dioxane (5 mL) and H2O (1 mL) were placed in a reaction vial. After N2 was bubbled through the mixture for 5 min, the mixture was heated and stirred at 90 °C for 1 h. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (20 mL) and washed with aqueous NH4Cl (20 mL). The organic phase was separated, dried over MgSO4, filtered and concentrated. The crude product was first purified by silica (20 g, 50-100% EtOAc in CH2Cl2) and then further purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.15% NH4OH, flow rate 60 mL / min) to give the desired products as a pair of diastereomers (white amorphous powder, combined yield 21%). Diastereomer 1. Peak 1.C 37 H 39 F5N9O2(M+H) + LCMS calculated m / z = 736.3; found 736.2. Diastereomer 2. Peak 2.C 37 H 39 F5N9O2(M+H) + LCMS calculated m / z = 736.3; found 736.2.

[0245] Step 2. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] To a reaction vial containing tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (diastereomer 1, 150 mg, 0.24 mmol) from step 1 was added TFA (1 mL) at room temperature. After stirring for 15 min, the volatiles were removed. The residue was redissolved in acetonitrile (2 mL) and cooled to 0 °C. DIPEA (0.36 mL) was added to the reaction, followed by (E)-4-fluorobut-2-enoic acid (42 mg, 0.41 mmol) and propylphosphonic anhydride solution (50% in EtOAc, 0.25 mL, 0.41 mmol). After stirring at 0° C. for 10 min, the reaction was quenched with aqueous NaHCO3 (5 mL), extracted with EtOAc (5 mL), washed with brine, and concentrated. The crude product was redissolved in acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give Example 1a (diastereomer 1) as a TFA salt in the form of a white amorphous powder (50 mg free base equivalent, 34% yield). C 36 H 34 F6N9O (M+H) + LCMS calculated m / z = 722.3; found 722.2. 1H NMR (500 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.87 (dd, J = 4.2, 1.7 Hz, 1H), 8.62 (dd, J = 8.5, 1.7 Hz, 1H), 8.41 (s, 1H), 7.82 (dd, J = 8.0, 5.8 Hz, 1H), 7.70 (dd, J = 8.5, 4.2 Hz, 1H), 7.65 (dd, J = 9.8, 8.0 Hz, 1H), 6.90 - 6.77 (m, 2H), 5.93 (s, 1H), 5.32 (s, 1H), 5.24 - 5.20 (m, 1H), 5.12 (dd, J = 3.6, 1.2 Hz, 1H), 4.97 (s, 1H), 4.76 (s, 1H), 4.57 (s, 1H), 4.37 - 4.21 (m, 2H), 3.61 (d, J = 12.7 Hz, 1H), 3.39 - 3.21 (m, 2H), 2.84 (s, 6H), 2.46 - 2.22 (m, 4H), 1.70 (s, 3H).

[0246] Example 1b (Diastereomer 2) was prepared by replacing tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (Diastereomer 1) from Step 1. C was prepared using the procedure described above using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (diastereomer 2) from step 1. 36 H 34 F6N9O (M+H) + LCMS calculated m / z = 722.3; found 722.2.

[0247] Example 2a and Example 2b. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0248] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] Intermediate 3 (100 mg, 0.15 mmol), (2-methoxy-3-methylphenyl)boronic acid (30 mg, 0.18 mmol), Pd(PPh3)4 (17 mg, 0.015 mmol), K3PO4 (95 mg, 0.45 mmol), dioxane (2.0 mL) and H2O (0.4 mL) were placed in a reaction vial. After bubbling N2 through the mixture for 5 min, the mixture was heated at 90 °C for 1 h. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (5.0 ml) and washed with aqueous NH4Cl (5.0 ml). The organic phase was separated, dried over MgSO4 and concentrated. The crude product was purified by silica (10 g, 50-100% EtOAc in DCM) to give the desired product as a pale yellow viscous oil (92 mg, 87% yield). C 36 H 43 F4N8O3(M+H) + LCMS calculated m / z = 711.3; found 711.2.

[0249] Step 2. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Example 1a and Step 2 of Example 1b, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. Example 2a. Diastereomer 1. Peak 1.C 35 H 38 F5N8O2(M+H) + LCMS calculated for: m / z = 697.3; found: 697.3. Example 2b. Diastereomer 2. Peak 2.C 35 H 38 F5N8O2(M+H) + LCMS calculated for: m / z = 697.3; found: 697.3.

[0250] Example 3a and Example 3b. 2-((2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0251] Step 1. tert-Butyl (2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Step 1 of Example 2a and Example 2b, using (3-chloro-2-methoxyphenyl)boronic acid instead of (2-methoxy-3-methylphenyl)boronic acid. 35 H 40 ClF4N8O2(M+H) + LCMS calculated for: m / z = 731.3; found: 731.2.

[0252] Step 2. 2-((2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in step 2 of Example 1a and Example 1b, using tert-butyl (2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. Example 3a. Diastereomer 1. Peak 1.C 34 H 35 ClF5N8O2(M+H) + LCMS calculated for: m / z = 717.3; found: 717.2. Example 3b. Diastereomer 2. Peak 2.C 34 H 35 ClF5N8O2(M+H) + LCMS calculated for: m / z = 717.3; found: 717.2.

[0253] Example 4a and Example 4b. 2-((2S,4S)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0254] Step 1. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate [ka] Intermediate 4 (3.50 g, 4.75 mmol) and methyl 2,2-difluoro-2-(fluorosulfonyl)-acetate (1.21 ml, 9.51 mmol) (MFDA), copper(I) iodide (0.272 g, 1.426 mmol), and NMP (20 mL) were placed in a sealed tube. The tube was flushed with N2, sealed, and heated at 80 °C overnight. The reaction was then cooled to room temperature, poured into NaHCO3, and extracted with EtOAc. The organic layers were combined, dried over MgSO4, and concentrated. The crude product was purified on silica (40 g, 0-50% EtOAc in hexanes) to give the desired product (2.5 g, 78% yield). C 27 H 33 BrF4N5O4S (M+H) + LCMS calculated for: m / z = 678.1; found: 678.3.

[0255] Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of intermediate 3, using tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate. C 23 H 24 BrF4N6O2S (M+H) + LCMS calculated for: m / z = 603.1; found: 603.2.

[0256] Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Example 1a and Step 1 of Example 1b, using tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate instead of intermediate 3. 32 H 29 F5N7O2S (M+H) + LCMS calculated for: m / z = 670.2; found: 670.2.

[0257] Step 4. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (85 mg, 0.13 mmol) was dissolved in CHCl (1 mL) and cooled to 0° C. mCPBA (33 mg, 0.19 mmol) was added in one portion and the reaction was stirred for 30 min before being quenched by the addition of saturated NaHCO (2 mL). The mixture was extracted with CHCl (5 mL). The combined organic layers were dried over MgSO, filtered, and concentrated to give a mixture of crude sulfoxide and sulfone. A vial containing (S)-1-((S)-1-methylpyrrolidin-2-yl)ethan-1-ol (16.4 mg, 0.13 mmol) and dry THF (1 mL) was cooled to 0° C. LiHMDS (0.13 mL, 1 M in THF) was added. After stirring for 10 min, the reaction was added dropwise to a solution of the crude sulfoxide in THF. After stirring for an additional 10 min, the reaction was quenched by the addition of saturated NH4Cl (5 mL) and extracted with EtOAc (15 mL). The combined organic layers were dried over MgSO4 and purified on silica (0-100% EtOAc in hexanes) to give the desired product as a white solid (75 mg, 79% yield). C 38 H 40 F5N8O3(M+H) + LCMS calculated for: m / z = 751.3; found: 751.5.

[0258] Step 5. 2-((2S,4S)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Example 1a and Step 2 of Example 1b, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. Example 4a. Diastereomer 1. Peak 1.C 37 H 35 F6N8O2(M+H) + LCMS calculated for: m / z = 737.3; found: 737.2. Example 4b. Diastereomer 2. Peak 2.C 37 H 35 F6N8O2(M+H) + LCMS calculated for: m / z = 737.3; found: 737.2.

[0259] Example 5a and Example 5b. 1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-1-yl)prop-2-en-1-one [ka]

[0260] Step 1. tert-Butyl 4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Step 1 of Example 4a and Example 4b, using Intermediate 5 instead of Intermediate 4. 21 H 23 BrF4N5O2S (M+H) + LCMS calculated for: m / z = 564.1; found: 564.0.

[0261] Step 2. tert-Butyl 4-(7-bromo-6-fluoro-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 4 of Example 4a and Example 4b, using tert-butyl 4-(7-bromo-6-fluoro-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-(methylthio)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. 27 H 34 BrF4N6O3(M+H) + LCMS calculated for: m / z = 645.2; found: 645.4.

[0262] Step 3. tert-Butyl 4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Example 1a and Step 1 of Example 1b, using tert-butyl 4-(7-bromo-6-fluoro-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of intermediate 3. 36 H 39 F5N7O3(M+H) + LCMS calculated for: m / z = 712.3; found: 712.5.

[0263] Step 4. 1-(4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-1-yl)prop-2-en-1-one [ka] tert-Butyl 4-(6-fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (25 mg, 0.039 mmol) was added to TFA (0.5 mL) and stirred at room temperature for 10 min. The solvent was removed under reduced pressure. The residue was dissolved in CHCl (1.0 mL) and cooled to 0° C. to which was added triethylamine (16 μL, 0.116 mmol), followed by acryloyl chloride (5.3 mg, 0.058 mmol) and the reaction was stirred at 0° C. for 20 min. The reaction was diluted with CHCl, washed with saturated NaHCO, the organic solvent was dried and concentrated. The crude product was redissolved in acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min). Example 5a. Diastereomer 1. Peak 1.C 34 H 33 F5N7O2(M+H) + LCMS calculated for: m / z = 666.3; found: 666.4. Example 5b. Diastereomer 2. Peak 2.C 34 H 33 F5N7O2(M+H) + LCMS calculated for: m / z = 666.3; found: 666.4.

[0264] Example 6. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0265] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] A mixture of intermediate 8 (400 mg, 0.728 mmol), (2,3-dimethylphenyl)boronic acid (164 mg, 1.092 mmol), potassium phosphate (464 mg, 2.184 mmol), and Pd(Ph3P)4 (84 mg, 0.073 mmol) in dioxane (6 mL) / water (1 mL) was evacuated and backfilled with nitrogen (this process was repeated a total of three times). The reaction was stirred at 105° C. for 1 h. The mixture was diluted with ethyl acetate and washed with water, brine. The organic layer was filtered, dried, and concentrated. The product was purified by column eluting with hexane / EtOAc (up to 80% EtOAc). C 31 H 36 FN6O2S (M+H) + LCMS calculated m / z = 575.3; found 575.3.

[0266] Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] To a solution of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (370.0 mg, 0.644 mmol) in CHCl (5.0 mL) at 0° C., m-CPBA (167 mg, 0.966 mmol) was added and the reaction was stirred for 10 min at this temperature. The reaction was quenched by the addition of saturated NaSO, diluted with ethyl acetate, washed with saturated NaHCO, brine, filtered, dried and concentrated. The crude material was used directly in the next step. C 31 H 36 FN6O3S (M+H) + LCMS calculated m / z = 591.3; found 591.3.

[0267] Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] To a solution of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (0.680 g, 1.121 mmol) and N,N,3-trimethylazetidin-3-amine (0.192 g, 1.681 mmol) was added triethylamine (0.625 mL, 4.48 mmol) and stirred at 70° C. for 2 h. The product was purified by column eluted with CHCl / MeOH (up to 15% MeOH). The atropisomers were separated by SFC (column, Phenomenex Lux 5um Cellulose-1, 21.2 x 250 mm, mobile phase 25% MeOH in CO2 at 70 mL / min isocratic) to give two peaks, designated PK1 and PK2. 36 H 46 FN8O2(M+H) + LCMS calculated m / z = 641.4; found 641.5.

[0268] Step 4. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was synthesized by the method described above using the method described above. This compound was synthesized by the method described above using the method described above. This compound was synthesized by the method described above. This compound was synthesized by the method described above. Prepared according to the procedure described in step 2 of Example 1a and Example 1b using methylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (PK1 from step 3) and using (E)-4-methoxybut-2-enoic acid instead of (E)-4-fluorobut-2-enoic acid. 36 H 44 FN8O2(M+H) + LCMS calculated m / z = 639.4; found 639.5. 1 H NMR (600 MHz, DMSO) δ 8.04 (s, 1H), 7.30 - 7.21 (m, 2H), 7.00 (dd, J = 7.4, 1.5 Hz, 1H), 6.81 - 6.70 (m, 2H), 5.30 (s, 1H), 4.96 (s, 1H), 4.74 (d, J = 13.8 Hz, 1H), 4.28 (d, J = 14.3 Hz, 1H), 4.11 (s, 2H), 3.64 (s, 1H), 3.39 (dd, J = 17.0, 8.5 Hz, 1H), 3.33 (s, 3H), 3.31 - 3.19 (m, 3H), 2.83 (s, 6H), 2.43 (s, 3H), 2.41 (m, 1H) 2.35 (s, 3H), 2.20 (s, 3H), 2.13 (d, J = 13.2 Hz, 1H), 1.94 (s, 3H), 1.70 (s, 3H).

[0269] Example 7. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0270] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 1 of Example 6, using 1-methyl-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole instead of (2,3-dimethylphenyl)boronic acid. 31 H 34 FN8O2S (M+H) + LCMS calculated m / z = 601.3; found 601.3.

[0271] Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of Example 6, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 31 H 34 FN8O3S (M+H) + LCMS calculated m / z = 617.2; found 617.4.

[0272] Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] To a solution of (S)-1-((S)-1-methylpyrrolidin-2-yl)ethan-1-ol (31.4 mg, 0.243 mmol) in THF (1.0 mL) at 0° C. was added LiHMDS (1.0 M in THF) (40.7 mg, 0.243 mmol) and stirred for 5 min. The formed solution was added to a solution of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (75.0 mg, 0.122 mmol) in THF (1.0 mL) at 0° C., and the reaction was then stirred at room temperature for 1 h. The mixture was diluted with ethyl acetate, washed with saturated NaHCO3, water, filtered, concentrated and used directly in the next step. 37 H 45 FN9O3(M+H) + LCMS calculated m / z = 682.4; found 682.5.

[0273] Step 4. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Example 1a and Step 2 of Example 1b, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 36 H 40 F2N9O2(M+H) + LCMS calculated m / z = 668.3; found 668.5. 1 H NMR (600 MHz, DMSO) δ 8.22 (s, 1H), 8.17 (s, 1H), 7.93 (d, J = 8.3 Hz, 1H), 7.72 (s, 1H), 7.13 (dd, J = 8.2 Hz, 1H), 6.92 - 6.79 (m, 2H), 5.87 (d, J = 16.1 Hz, 1H), 5.58 (d, J = 7.5 Hz, 1H), 5.31 (d, J = 6.4 Hz, 1H), 5.21 (d, J = 2.4 Hz, 1H), 5.14 (d, J = 2.9 Hz, 2H), 5.00 (s, 1H), 4.76 (d, J = 13.8 Hz, 1H), 4.32 (d, J = 14.2 Hz, 1H), 4.11 (s, 3H), 3.96 (p, J = 7.7 Hz, 1H), 3.65- 3.59 (m, 3H), 3.45 - 3.23 (m, 2H), 3.19 (m,1H), 3.14 (s, 3H), 2.44 (s, 3H), 2.32- 2.16 (m, 2H), 2.11- 1.89 (m, 2H), 1.59 (s, 3H).

[0274] Example 8. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0275] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Step 1 of Example 6, using (6-methylpyridin-3-yl)boronic acid instead of (2,3-dimethylphenyl)boronic acid. 29 H 33 FN7O2S (M+H) + LCMS calculated m / z = 562.2; found 562.3.

[0276] Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of Example 6, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. 29 H 33 FN7O3S (M+H) + LCMS calculated m / z = 578.2; found 578.4.

[0277] Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 3 of Example 7, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 35 H 44 FN8O3(M+H) +LCMS calculated m / z = 643.4; found 643.5.

[0278] Step 4. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in step 2 of Example 1a and Example 1b, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 34 H 39 F2N8O2(M+H) + LCMS calculated m / z = 629.3; found 629.5.

[0279] Example 9. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0280] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 1 of Example 6, using 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole instead of (2,3-dimethylphenyl)boronic acid. 31 H 34 FN8O2S (M+H) + LCMS calculated m / z = 601.3; found 601.1.

[0281] Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of Example 6, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 31 H34 FN8O3S (M+H) + LCMS calculated m / z = 617.3; found 617.3.

[0282] Step 3. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 3 of Example 7, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 37 H 45 FN9O3(M+H) + LCMS calculated m / z = 682.4; found 682.4

[0283] Step 4. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in step 4 of Example 6, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 37 H 43 FN9O3(M+H) + LCMS calculated m / z = 680.4; found 680.3.

[0284] Example 10. 2-((2S,4S)-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in steps 1-4 of Example 9, using (4-fluorophenyl)boronic acid instead of 1-methyl-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole. 35 H 40 F2N7O3(M+H) + LCMS calculated m / z = 644.3; found 644.4. 1H NMR (500 MHz, DMSO) δ 10.09 - 9.21 (s, 1H), 8.51 - 7.69 (s, 1H), 7.53 - 7.44 (dd, J = 8.5, 5.7 Hz, 2H), 7.44 - 7.36 (t, J = 8.9 Hz, 2H), 6.83 - 6.63 (m, 2H), 5.92 - 5.72 (m, 1H), 5.66 - 5.47 (m, 1H), 5.41-4.20 (m, 2H), 4.18 - 4.03 (d, J = 2.5 Hz, 2H), 4.00 - 3.85 (m, 1H), 3.69 - 3.53 (m, 2H), 3.39 - 3.32 (s, 3H), 3.27 - 3.10 (m, 5H), 2.55 - 2.36 (d, J = 13.8 Hz, 6H), 2.38 - 2.23 (m, 2H), 2.19 - 2.04 (m, 2H), 2.00 - 1.88 (m, 2H), 1.64 - 1.50 (d, J = 6.1 Hz, 3H).

[0285] Example 11a and Example 11b. 8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka]

[0286] Step 1. tert-Butyl 4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of intermediate 8, using intermediate 5 instead of tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate. 21 H 26 BrFN5O2S (M+H) + LCMS calculated m / z = 510.1; found 510.1.

[0287] Step 2. tert-Butyl 4-(7-bromo-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of Example 6, using tert-butyl 4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. 21 H 26 BrFN5O3S (M+H-tBu) + LCMS calculated m / z = 470.0; found 470.0.

[0288] Step 3. tert-Butyl 4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] To a solution of tert-butyl 4-(7-bromo-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (2.51 g, 4.7 mmol) in THF (16 mL) was added (S)-1-((S)-1-methylpyrrolidin-2-yl)ethan-1-ol (1.23 g, 9.54 mmol) and DBU (1.44 mL, 9.54 mmol). The mixture was stirred at room temperature overnight, then diluted with EtOAc and extracted with saturated NH4Cl. The combined organic layers were dried over MgSO4, filtered, concentrated and purified by column chromatography (0-8% MeOH / DCM). C 27 H 37 BrFN6O3(M+H) + LCMS calculated m / z = 591.2; found 591.2.

[0289] Step 4. 8-(6-Fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1-(piperidin-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka] tert-Butyl 4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (1.56 g, 2.64 mmol), 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphthonitrile (1.472 g, 5.27 mmol), SPhos Pd G4 (315 mg, 0.396 mmol), K3PO4 (1.679 g, 7.91 mmol), as well as dioxane (11 mL) and H2O (2.34 mL) were placed in a reaction vial. N2 was bubbled through the mixture for 5 min, and then the mixture was heated at 100 °C for 3 h. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (20 mL) and washed with aqueous NH4Cl (20 mL). The organic phase was separated, dried over MgSO4, filtered and then concentrated. The concentrated residue was redissolved in CH2Cl2 (10 mL) and TFA (5 mL) and stirred at room temperature for 30 min. The solvent was then removed and the crude product was purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product as a TFA salt. 33 H 35 FN7O (M+H) + LCMS calculated m / z = 564.3; found 564.2.

[0290] Step 5. 8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka] To a solution of 8-(6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1-(piperidin-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile in CHCl (0.4 M) was added triethylamine (5 eq.), followed by a solution of acryloyl chloride (1.5 eq.) in CHCl (0.5 M). The mixture was stirred at room temperature for 30 min and then quenched by adding MeOH. The mixture was concentrated under reduced pressure, diluted with AcN, and purified by preparative LCMS. Example 11a. Diastereomer 1. Peak 1.C 36 H 37 FN7O2(M+H) + LCMS calculated m / z = 618.3; found 618.2. Example 11b. Diastereomer 2. Peak 2.C 36 H 37 FN7O2(M+H) + LCMS calculated m / z = 618.3; found 618.2. 1H NMR (600 MHz, DMSO) δ 9.98 - 9.50 (d, J = 10.7 Hz, 1H), 8.56 - 8.46 (dd, J = 8.5, 1.3 Hz, 1H), 8.38 - 8.24 (m, 2H), 8.16 - 8.10 (dd, J = 7.2, 1.4 Hz, 1H), 7.91 - 7.83 (dd, J = 8.3, 7.0 Hz, 1H), 7.81 - 7.72 (dd, J = 8.3, 7.2 Hz, 1H), 7.71 - 7.64 (d, J = 7.0 Hz, 1H), 6.98 - 6.84 (dd, J = 16.7, 10.5 Hz, 1H), 6.25 - 6.16 (dd, J = 16.8, 2.4 Hz, 1H), 5.85 - 5.77 (m, 1H), 5.77 - 5.73 (dd, J = 10.5, 2.4 Hz, 1H), 5.57 - 5.44 (m, 1H), 4.72 - 4.24 (m, 1H), 4.01 - 3.88 (m, 1H), 3.71 - 3.48 (m, 2H), 3.31 - 3.19 (m, 2H), 3.19 - 3.11 (d, J = 4.8 Hz, 3H), 2.48 - 2.34 (m, 2H), 2.34 - 2.03 (m, 9H), 2.00 - 1.86 (m, 2H), 1.64 - 1.52 (d, J = 6.1 Hz, 3H).

[0291] Example 12. 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0292] Step 1. tert-Butyl (2S,4S)-4-((7-bromo-2-chloro-8-fluoro-6-iodo-3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate [ka] To a solution of 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline (Intermediate 1) (7.9 g, 16.96 mmol) and tert-butyl (2S,4S)-4-amino-2-(cyanomethyl)piperidine-1-carboxylate (Intermediate 7) (4.87 g, 20.35 mmol) in dioxane (50 mL) at room temperature was added DIPEA (4.5 mL, 25.4 mmol). After the mixture was stirred at room temperature overnight, most of the dioxane was removed under reduced pressure. To the residue was added ice and water, and the slurry was stirred vigorously. The solid precipitate was collected via filtration and air-dried to give the desired product as a yellow solid (quantitative yield). C 21 H 22 BrClFIN5O4(M+H) + LCMS calculated for: m / z = 668.0; found: 667.8.

[0293] Step 2. tert-Butyl (2S,4S)-4-((7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-8-fluoro-6-iodo-3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate [ka] To a solution of tert-butyl (2S,4S)-4-((7-bromo-2-chloro-8-fluoro-6-iodo-3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate (2.5 g, 3.74 mmol) in dioxane (20 mL) was added DIPEA (2.61 mL, 14.95 mmol) and N,N,3-trimethylazetidin-3-amine dihydrochloride (1.05 g, 5.61 mmol). The mixture was stirred at 50° C. overnight, after which most of the dioxane was removed under reduced pressure. Ice and water were added to the residue, and the slurry was stirred vigorously until a fine powder was formed. The solid precipitate was collected via filtration and air-dried to give the desired product (2.7 g, 97% yield). C 27 H 35 BrFIN7O4(M+H) + LCMS calculated for: m / z = 746.1; found: 746.1.

[0294] Step 3. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-iodo-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] A mixture of tert-butyl (2S,4S)-4-((7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-8-fluoro-6-iodo-3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate (1.0 g, 1.34 mmol) and iron (450 mg, 8.04 mmol) was dissolved in AcOH (5.0 mL) and heated at 60° C. until complete reduction of the nitro group. The mixture was cooled to room temperature, then diluted with EtOAc and filtered through a pad of Celite. The filtrate was concentrated to give tert-butyl (2S,4S)-4-((3-amino-7-bromo-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-8-fluoro-6-iodoquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate. C 27 H 37 BrFIN7O2(M+H) + LCMS calculated for: m / z = 716.1; found: 716.1.

[0295] To the concentrated residue, triethyl orthoformate (0.45 mL, 2.68 mmol) and toluene (10 mL) were added. The mixture was heated at 100° C. overnight, then cooled to room temperature and concentrated. The residue was purified by column chromatography (0-10% MeOH / CH2Cl2) to give the desired product (534 mg, 55% yield). 28 H 35 BrFIN7O2(M+H) + LCMS calculated for: m / z = 726.1; found: 726.1.

[0296] Step 4. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of intermediate 8, using tert-butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-iodo-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate. 29 H 38 BrFN7O2(M+H) + LCMS calculated m / z = 614.2; found 614.3.

[0297] Step 5. tert-Butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] A mixture of tert-butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (785 mg, 1.277 mmol), (2-chloro-3-methylphenyl)boronic acid (283 mg, 1.661 mmol), sodium carbonate (542 mg, 5.11 mmol), and Pd(Ph3P)4 (221 mg, 0.192 mmol) in dioxane (5.5 mL) / water (1.1 mL) was evacuated and backfilled with nitrogen (this process was repeated a total of three times). The reaction was stirred at 90° C. for 3 h. The mixture was diluted with ethyl acetate and washed with water, brine. The organic layer was filtered, dried over MgSO4, concentrated and purified by preparative LCMS. 36 H44 ClFN7O2(M+H) + LCMS calculated m / z = 660.3; found 660.3.

[0298] Step 6. 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in step 2 of Example 1a and Example 1b, using tert-butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. The product was isolated as a mixture of diastereomers. C 35 H 39 ClF2NO (M+H) + LCMS calculated m / z = 646.3; found 646.3. 1H NMR (600 MHz, DMSO) δ 10.66 - 10.31 (s, 1H), 8.54 - 8.50 (m, 1H), 7.96 - 7.82 (d, J = 4.8 Hz, 1H), 7.51 - 7.46 (m, 1H), 7.44 - 7.37 (td, J = 7.6, 2.4 Hz, 1H), 7.29 - 7.17 (d, J = 7.6 Hz, 1H), 6.89 - 6.80 (m, 2H), 5.60 - 5.39 (m, 1H), 5.36 - 4.85 (m, 1H), 5.24 - 5.11 (dd, J = 46.5, 2.5 Hz, 2H), 4.79 - 4.19 (m, 5H), 3.72 - 3.56 (m, 1H), 3.49 - 3.14 (m, 2H), 2.85 - 2.75 (s, 6H), 2.49 - 2.03 (m, 10H), 1.71 - 1.61 (s, 3H).

[0299] Example 13. 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0300] Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedures described in step 1 and then step 3 of Example 12. 22 H 22 BrClFIN5O2(M+H) +LCMS calculated m / z = 648.0; found 648.0.

[0301] Step 2. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of intermediate 4, using tert-butyl (2S,4S)-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-4-(7-bromo-4-chloro-6-fluoro-8-iodo-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate. 23 H 25 BrFIN5O2S (M+H) + LCMS calculated for: m / z = 660.0; found: 660.0.

[0302] Step 3. tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 2 of intermediate 8, using tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate. 24 H 28 BrFN5O2S (M+H) + LCMS calculated m / z = 548.1; found 548.1.

[0303] Step 4. tert-Butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in step 5 of Example 12, using tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate. 31 H 34 ClFN5O2S (M+H) + LCMS calculated m / z = 594.2; found 594.3.

[0304] Step 5. tert-Butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in steps 2-3 of Example 7, using tert-butyl (2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. C 37 H 45 ClFN6O3(M+H) + LCMS calculated m / z = 675.3; found 675.3.

[0305] Step 6. 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka] This compound was synthesized by the method of the present invention by using tert-butyl (2S,4S)-4-(7-(2-chloro)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. The compound was prepared according to the procedure described in step 2 of Example 1a and Example 1b, using (E)-4-fluorobut-2-enoic acid instead of 2-fluoroacrylic acid, and using (3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate. The product was isolated as a mixture of diastereomers. 35 H 38 ClF2N6O2(M+H) + LCMS calculated m / z = 647.3; found 647.3.

[0306] Examples 14a and 14b. 8-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka]

[0307] Step 1. tert-Butyl (2R,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Intermediate 4, using tert-butyl (2R,4S)-4-amino-2-methylpiperidine-1-carboxylate instead of tert-butyl (2S,4S)-4-amino-2-(2-(tert-butoxy)-2-oxoethyl)piperidine-1-carboxylate. 21 H 25 BrFIN5O2S (M+H) + LCMS calculated for: m / z = 636.0; found: 636.0.

[0308] Step 2. tert-Butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate [ka] A mixture of tert-butyl (2R,4S)-4-(7-bromo-6-fluoro-8-iodo-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate (5 g, 7.86 mmol), methylboronic acid (0.941 g, 15.72 mmol), potassium phosphate (5.00 g, 23.57 mmol), and dichlorobis(triphenylphosphine)-palladium(II) (0.827 g, 1.179 mmol) in dioxane (60 mL) and water (20 mL) was heated to 90° C. and stirred at 90° C. for 3 h. The mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with AcOEt, and the combined organic layers were washed with brine, dried over Na2SO4, filtered, and evaporated. The residue was purified by column chromatography (0-20% EtOAc in CH2Cl2) to give the desired product as a brown solid. 22 H 28 BrFN5O2S (M+H) + LCMS calculated m / z = 524.1; found 524.1.

[0309] Step 3. tert-Butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in steps 2-3 of Example 7, using tert-butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate. The residue was purified by column chromatography (20-80% AcOEt in DCM) to give the desired product as a brown solid. C 28 H 39 BrFN6O3(M+H) + LCMS calculated m / z = 605.2; found 605.2.

[0310] Step 4. tert-Butyl (2R,4S)-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate [ka] A mixture of tert-butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate (200 mg, 0.330 mmol), 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphthonitrile (184 mg, 0.661 mmol), tripotassium phosphate (351 mg, 1.651 mmol), and SPhos Pd G4 (79 mg, 0.099 mmol) in dioxane (8 mL) and water (2 mL) was heated to 90 °C. After stirring at the same temperature for 3 h, the mixture was cooled to room temperature, diluted with EtOAc and water, and separated. The aqueous layer was extracted with AcOEt, and the combined organic layers were washed with brine, dried over Na2SO4, filtered, and evaporated. The residue was purified by column chromatography (20-80% EtOAc in CH2Cl2) to give the desired product as a brown solid. 39 H 45 FN7O3(M+H) + LCMS calculated m / z = 678.4; found 678.4.

[0311] Step 5. 8-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka] tert-Butyl (2R,4S)-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate (400 mg, 0.578 mmol) was dissolved in 5 mL of TFA. The mixture was stirred at room temperature for 10 minutes, and then the solvent was removed.

[0312] The residue was dissolved in acetonitrile (10 mL) and Et3N (484 μl, 3.47 mmol) was added. After stirring at 0° C. for several minutes, the cloudy mixture became a clear solution. Acryloyl chloride (94 μl, 1.157 mmol) was added. The mixture was stirred at 0° C. for 30 minutes, then diluted with TFA and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product. Example 14a. Diastereomer 1. Peak 1.C 37 H 39 FN7O2(M+H) + LCMS calculated m / z = 632.3; found 632.3. Example 14b. Diastereomer 2. Peak 2.C 37 H 39 FN7O2(M+H) + LCMS calculated m / z = 632.3; found 632.3. 1H NMR (500 MHz, DMSO) δ 9.87 (s, 1H), 8.51 (dd, J = 8.5, 1.4 Hz, 1H), 8.31 (dd, J = 8.3, 1.3 Hz, 1H), 8.21 (s, 1H), 8.14 (dd, J = 7.2, 1.3 Hz, 1H), 7.87 (dd, J = 8.3, 7.1 Hz, 1H), 7.77 (dd, J = 8.3, 7.2 Hz, 1H), 7.68 (d, J = 7.0 Hz, 1H), 6.93 - 6.87 (m, 1H), 6.16 (dd, J = 16.7, 2.4 Hz, 1H), 5.95 (s, 1H), 5.73 (dd, J = 10.5, 2.4 Hz, 1H), 5.51 (dd, J = 10.1, 5.9 Hz, 1H), 5.08 (s, 1H), 4.68 (s, 1H), 4.27 (d, J = 13.9 Hz, 1H), 3.95 (dd, J = 7.8, 7.7 Hz, 1H), 3.68 (s, 1H), 3.57 (dq, J = 12.2, 6.2 Hz, 1H), 3.17 (m, J = 4.9 Hz, 3H), 2.46 (d, J = 9.7 Hz, 2H), 2.35 - 2.26 (m, 2H), 2.22 (s, 3H), 2.10 (td, J = 11.8, 5.2 Hz, 1H), 1.93 (tq, J = 14.3, 6.8 Hz, 2H), 1.58 (d, J = 6.1 Hz, 4H), 1.51 (d, J = 7.0 Hz, 2H).

[0313] Examples 15a and 15b. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0314] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] A mixture of intermediate 8 (1 g, 1.755 mmol), 5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (0.938 g, 2.63 mmol), potassium phosphate tribasic (1.862 g, 8.77 mmol), and Pd(Ph3P)4 (0.203 g, 0.175 mmol) in dioxane (12 mL) and water (6 mL) was heated to 100° C. and stirred at 100° C. for 2 h. The mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc, and the combined organic layers were washed with brine, dried over Na2SO4, filtered, and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH2Cl2) to give the desired product as a brown solid. 37 H 44 FN8O3S (M+H) + LCMS calculated m / z = 699.3; found 699.3.

[0315] Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was synthesized by the method of the present invention by replacing tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate in step 2 with tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pi Prepared according to the procedure described in steps 2-3 of example 6 using N-ethyl-N-methylazetidin-3-amine dihydrochloride instead of N,N,3-trimethylazetidin-3-amine in step 3, using N-lan-2-yl-1H-indazol-4-yl-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. The crude residue was purified by column chromatography (20-80% EtOAc in CH2Cl2) to give the desired product as a brown solid. C 42 H 54 FN 10 O3 (M+H) + LCMS calculated m / z = 765.4; found 765.3.

[0316] Step 3. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka] tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (378 mg, 0.494 mmol) was dissolved in TFA (5 mL). The mixture was stirred at 60° C. for 10 min, and then the solvent was removed. The residue was mixed with 2-fluoroacrylic acid (131 mg, 1.455 mmol), followed by the addition of acetonitrile (10 mL), followed by the addition of DIPEA (506 μl, 2.89 mmol) and T3P (853 μl, 50% in EtOAc, 1.447 mmol). After stirring at room temperature overnight, the mixture was diluted with TFA, filtered and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product as a TFA salt. Example 15a. Diastereomer 1. Peak 1.C 35 H 39 F2N 10 O (M+H) + LCMS calculated m / z = 653.3; found 653.3. 1H NMR (600 MHz, DMSO) δ 13.02 (s, br, 1H), 10.21 (s, br, 1H), 8.09 (s, 1H), 7.47 (s, 1H), 7.30 (s, 1H), 5.86 (td, J = 11.4, 5.5 Hz, 1H), 5.45 - 5.38 (m, 1H), 5.33 (m, 1H), 4.69 (s, 8H), 4.40 (d, J = 8.8 Hz, 1H), 4.15 (s, 1H), 3.73 (s, 1H), 3.36 (dd, J = 17.2, 6.6 Hz, 2H), 3.11 (m, 1H), 2.86 (s, 3H), 2.49 (d, J = 9.5 Hz, 1H), 2.47 (s, 3H), 2.13 (s, 3H), 2.02 (s, 3H), 1.25 (t, J = 7.3 Hz, 3H). Example 15b. Diastereomer 2. Peak 2.C 35 H 39 F2N 10 O (M+H) + LCMS calculated m / z = 653.3; found 653.3.

[0317] Example 16a and Example 16b. 8-(6-fluoro-1-(1-((E)-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka] A round bottom flask was charged with 8-(6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1-(piperidin-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile (600 mg, 0.854 mmol) from steps 1-4 of Example 11, propylphosphonic anhydride solution (50% in EtOAc, 1.6 mL, 2.56 mmol), (E)-4-fluorobut-2-enoic acid (267 mg, 2.56 mmol), and acetonitrile (15 mL). The reaction flask was cooled to 0° C. and TEA (1.7 mL, 12.8 mmol) was added to the reaction. After stirring at room temperature for 2 h, the reaction was quenched with aqueous NaHCO3 (10 mL), extracted with EtOAc (25 mL x 3) and concentrated. The crude mixture was redissolved in acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product. Example 16a. Diastereomer 1. Peak 1.C 37 H 38 F2N7O2(M+H) + LCMS calculated for: m / z = 650.3; found: 650.3. Example 16b. Diastereomer 2. Peak 2.C 37 H 38 F2N7O2(M+H) + LCMS calculated for: m / z = 650.3; found: 650.3.

[0318] Example 17a and Example 17b. 8-(1-((2S,4S)-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka]

[0319] Step 1. tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in steps 2-3 of Example 6, using intermediate 8 instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. The crude residue was purified on silica (0-10% MeOH in DCM) to give the desired product as a brown solid (700 mg, 63% yield). 28 H 37 BrFN8O2(M+H) + LCMS calculated for: m / z = 615.2; found: 615.2.

[0320] Step 2. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(8-cyanonaphthalen-1-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] tert-Butyl (2S,4S)-4-(7-bromo-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (500 mg, 0.79 mmol), SPhos Pd G4 (97 mg, 0.12 mmol), 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphthonitrile (453 mg, 1.63 mmol), K3PO4 (571 mg, 2.44 mmol), dioxane (9 mL) and H2O (3 mL) were placed in a reaction vial. N2 was bubbled through the mixture for 5 min, and then the mixture was heated at 90 °C for 2 h. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (30 mL) and washed with water (20 mL). The organic phase was separated, dried over MgSO4 and purified on silica (0-10% MeOH in DCM) to give the desired product as a brown solid (358 mg, 64% yield). 39 H 43 FN9O2(M+H) + LCMS calculated for: m / z = 688.4; found: 688.3.

[0321] Step 3. 8-(1-((2S,4S)-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile [ka] This compound is tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. Prepared according to the procedure described in Example 1a and step 2 of Example 1b using (7-(8-cyanonaphthalen-1-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate and using 2-fluoroacrylic acid instead of (E)-4-fluorobut-2-enoic acid. Example 17a. Diastereomer 1. Peak 1.C 37 H 36 F2N9O (M+H) + LCMS calculated for: m / z = 660.3; found: 660.3. 1 H NMR (600 MHz, DMSO) δ 8.48 (dd, J = 8.4, 1.3 Hz, 1H), 8.28 (dd, J = 8.4, 1.3 Hz, 1H), 8.12 (dd, J = 7.1, 1.3 Hz, 1H), 8.06 (s, 1H), 7.85 (dd, J = 8.3, 7.1 Hz, 1H), 7.75 (dd, J = 8.3, 7.1 Hz, 1H), 7.62 (dd, J = 7.1, 1.3 Hz, 1H), 5.90-5.75 (m, 1H), 5.45-5.30 (m, 2H), 4.00-4.50 (m, 11H), 2.85 (s, 6H), 2.50 (s, 1H), 2.20-2.15 (m, 4H), 1.70 (s, 3H). Example 17b. Diastereomer 2. Peak 2.C 37 H 36 F2N9O (M+H) + LCMS calculated for: m / z = 660.3; found: 660.3.

[0322] Examples 18a and 18b. 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0323] Step 1. 7-Bromo-6,8-dichloro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione [ka] To a solution of 2-amino-4-bromo-3-chlorobenzoic acid (10.0 g, 39.9 mmol) in 1,4-dioxane (100 mL) was added NCS (5.6 g, 41.9 mmol) and the reaction was stirred at 70° C. overnight. After the mixture was cooled to room temperature, triphosgene (4.7 g, 15.96 mmol) was added and then stirred at 80° C. for 2 hours. Upon completion, ice was added and the mixture was stirred for 30 minutes. The solid precipitate was collected via filtration and washed with ice water to give the desired product.

[0324] Step 2. 7-Bromo-6,8-dichloro-3-nitroquinoline-2,4-diol [ka] To a solution of ethyl 2-nitroacetate (2.66 g, 19.97 mmol) in toluene (40.0 mL) at room temperature was added DIPEA (3.49 mL, 19.97 mmol). After stirring for 10 min, 7-bromo-6,8-dichloro-2H-benzo[d][1,3]oxazine-2,4(1H)-dione (3.1 g, 10.0 mmol) was added and the resulting mixture was stirred at 95 °C for 3 h. After cooling the reaction with ice water, the solid was collected via filtration and washed with a small amount of ethyl acetate to give the desired product as a yellow solid (1.05 g, 30%). C9H4BrCl2N2O4 (M+H) + LCMS calculated for: m / z = 352.9; found 353.0.

[0325] Step 3. 7-Bromo-2,4,6,8-tetrachloro-3-nitroquinoline [ka] To a mixture of 7-bromo-6,8-dichloro-3-nitroquinoline-2,4-diol (1.0 g, 2.82 mmol) in POCl3 (6.0 mL, 64.4 mmol) at room temperature was added DIPEA (0.98 mL, 5.6 mmol). The reaction was then stirred at 100 °C for 3 h. The solvent was removed under reduced pressure. The concentrated residue was redissolved in ethyl acetate and then washed with aqueous NaHCO3. The combined organic layers were concentrated under reduced pressure to give the crude product, which was used directly in the next step without further purification.

[0326] Step 4. tert-Butyl (2S,4S)-4-((7-bromo-6,8-dichloro-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate [ka] To a solution of 7-bromo-2,4,6,8-tetrachloro-3-nitroquinoline (1 g, 2.56 mmol) in CHCl (10 mL) was added DIPEA (1.8 mL, 10.24 mmol) and Intermediate 7 (730 mg, 3.0 mmol). After stirring the mixture at room temperature overnight, N,N,3-trimethylazetidin-3-amine (350 mg, 3.0 mmol) was added and the mixture was stirred at room temperature until completion. The reaction was diluted with CHCl and then washed with saturated NHCl solution. The solvent was removed under reduced pressure and the crude product was used directly in the next step without further purification. C 27 H 35 BrCl2N7O4(M+H) + LCMS calculated for: m / z = 670.1; found 670.1.

[0327] Step 5. tert-Butyl (2S,4S)-4-(7-bromo-6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] A slurry of tert-butyl (2S,4S)-4-((7-bromo-6,8-dichloro-2-(3-(dimethylamino)-3-methylazetidin-1-yl)-3-nitroquinolin-4-yl)amino)-2-(cyanomethyl)piperidine-1-carboxylate (1.7 g, 2.56 mmol), iron (1.43 g, 25.6 mmol), and NH4Cl (1.37 g, 25.6 mmol) in THF / MeOH / H2O (1:1:1) (30 mL) was heated at 65° C. for 30 min. Upon completion, the reaction was diluted with MeOH and filtered. The solvent was removed under reduced pressure, and the residue was basified with NaHCO3 and extracted with CH2Cl2. The solvent was removed under reduced pressure.

[0328] The concentrated residue was redissolved in acetic acid (5 mL) and sodium nitrite (176 mg, 5.12 mmol) was added. The mixture was stirred at room temperature for 1 h. Upon completion, the solvent was removed under reduced pressure and the residue was basified with NaHCO3 and extracted with CH2Cl2. The combined organic layers were dried over MgSO4, filtered, concentrated to dryness, and purified on silica gel to give the desired product (1.0 g, 60%). 27 H 34 BrCl2N8O2(M+H) + LCMS calculated for: m / z = 651.1; found 651.0.

[0329] Step 6. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] To a solution of tert-butyl (2S,4S)-4-(7-bromo-6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (650 mg, 1.0 mmol) in dioxane / HO=4:1 (10 mL) was added KPO (850 mg, 4 mmol), (5-fluoroquinolin-8-yl)boronic acid (286 mg, 1.5 mmol), and Pd(PPh) (80 mg, 0.07 mmol) and the reaction mixture was heated to 90 °C for 1.5 h. Upon completion, the reaction was diluted with water, extracted with CH2Cl2, and the combined organic layers were dried over MgSO4, filtered, concentrated to dryness, and purified on silica gel to give the desired product as a mixture of diastereomers (550 mg, 76%). 36 H 39 Cl2FN9O2(M+H) + LCMS calculated for: m / z = 718.3; found 718.2.

[0330] Step 7. 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in step 4 of example 6, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. The product was purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.15% NH4OH, flow rate 60 mL / min) to give the desired product. Example 18a. Diastereomer 1. Peak 1.C 36 H 37 Cl2FN9O2(M+H) + LCMS calculated m / z = 716.2; found 716.2. Example 18b. Diastereomer 2. Peak 2.C 36 H 37 Cl2FN9O2(M+H) + LCMS calculated m / z = 716.2; found 716.2.

[0331] Examples 19a and 19b. 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Example 1a and Step 2 of Example 1b, using tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. Example 19a. Diastereomer 1. Peak 1.C 35 H 34 Cl2F2NO (M+H) + LCMS calculated m / z = 704.2; found 704.2. Example 19b. Diastereomer 2. Peak 2.C 35 H 34 Cl2F2NO (M+H) + LCMS calculated m / z = 704.2; found 704.2.

[0332] Example 20. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedures described in Example 1a and Example 1b (Step 2 for Example 1a), using (E)-4-methoxybut-2-enoic acid instead of (E)-4-fluorobut-2-enoic acid.

[0333] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] Intermediate 3 (1.3 g, 1.9 mmol), XPhos Pd G2 (76 mg, 0.097 mmol), (5-fluoroquinolin-8-yl)boronic acid (408 mg, 2.1 mmol), K3PO4 (1.24 g, 5.83 mmol), dioxane (5 mL) and H2O (1 mL) were placed in a reaction vial. N2 was bubbled through the mixture for 5 min, and then the mixture was heated at 90 °C for 1 h. Upon completion, the reaction was cooled to room temperature, diluted with EtOAc (20 mL) and washed with aqueous NH4Cl (20 mL). The organic phase was separated, dried over MgSO4, filtered and concentrated. The crude product was first purified by silica (20 g, 50-100% EtOAc in CH2Cl2) and then further purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.15% NH4OH, flow rate 60 mL / min) to separate the diastereomers (white amorphous powder, combined yield 21%). Diastereomer 1. Peak 1.C 37 H 39 F5N9O2(M+H) + LCMS calculated m / z = 736.3; found 736.2. Diastereomer 2. Peak 2.C 37 H 39 F5N9O2(M+H) + LCMS calculated m / z = 736.3; found 736.2.

[0334] Step 2. 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile To a reaction vial containing tert-butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (diastereomer 1, 300 mg, 0.41 mmol) from step 1 was added TFA (1 mL) at room temperature. After stirring for 15 min, the volatiles were removed. The residue was redissolved in acetonitrile (4 mL) and cooled to 0 °C. DIPEA (0.21 mL) was added to the reaction followed by (E)-4-methoxybut-2-enoic acid (71 mg, 0.61 mmol) and propylphosphonic anhydride solution (50% in EtOAc, 0.50 mL, 0.82 mmol). After stirring at 0° C. for 10 min, the reaction was quenched with aqueous NaHCO3 (5 mL), extracted with EtOAc (5 mL), washed with brine, and concentrated. The crude product was redissolved in acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give Example 20 (single diastereomer) as a TFA salt in the form of a white amorphous powder. 37 H 37 F5N9O2(M+H) + LCMS calculated m / z = 734.3; found 734.4. 1H NMR (600 MHz, DMSO-d6) δ 10.80 (s, 1H), 8.87 (dd, J = 4.2, 1.7 Hz, 1H), 8.62 (dd, J = 8.5, 1.8 Hz, 1H), 8.41 (s, 1H), 7.82 (dd, J = 8.0, 5.9 Hz, 1H), 7.70 (dd, J = 8.5, 4.1 Hz, 1H), 7.65 (dd, J = 9.7, 8.0 Hz, 1H), 6.81 - 6.72 (m, 2H), 5.93 (s, 1H), 5.31 (s, 1H), 4.96 (m, 2H), 4.76 - 4.32 (m, 2H), 4.11 (d, J = 3.1 Hz, 2H), 3.67 - 3.34 (m, 2H), 3.33 (s, 3H), 3.19 (td, J = 16.6, 10.1 Hz, 2H), 2.84 (s, 6H), 2.49 - 2.43 (m, 2H), 2.33 (s, 1H), 2.21 (d, J = 15.7 Hz, 1H), 1.71 (s, 3H).

[0335] Example 21. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0336] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (315 mg, 0.5 mmol), bis(pinacolato)diboron (165 mg, 0.65 mmol), Pd(dppf)Cl2 (36 mg, 0.05 mmol), potassium acetate (147 mg, 1.5 mmol), and dioxane (5 mL) were placed in a reaction vial. After bubbling N2 through the mixture for 5 min, the mixture was heated at 80 °C for 16 h. Upon completion, the reaction was cooled to room temperature, diluted with DCM (100 mL) and washed with aqueous NH4Cl (20 mL). The organic phase was separated, dried over MgSO4, filtered and concentrated. The crude product was used in the next step without further purification.

[0337] Step 2. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] The crude tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate from step 1 immediately preceding, 1-chloro-4-fluoroisoquinoline (118 mg, 0.65 mmol), Pd(PPh3)4 (58 mg, 0.05 mmol), K3PO4 (318 mg, 1.5 mmol), dioxane (4 mL), and water (1 mL) were placed in a reaction vial. The mixture was heated at 90° C. for 1 h. Upon completion, the reaction was cooled to room temperature, diluted with ethyl acetate (100 mL) and washed with aqueous NH4Cl (20 mL). The organic phase was separated, dried over MgSO4, filtered and concentrated.

[0338] The crude product was dissolved in 5 mL of DCM / TFA (1:1) solution. Upon completion, the reaction was concentrated. The crude product was redissolved in acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product as a mixture of diastereomers. 33 H 35 F2N8O (M+H) + LCMS calculated m / z = 597.3; found 597.3.

[0339] Step 3. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile (30 mg, 0.05 mmol), DIPEA (0.087 mL, 0.5 mmol), (E)-4-methoxybut-2-enoic acid (12 mg, 0.1 mmol), 1 mL MeCN, and propylphosphonic anhydride solution (50% in EtOAc, 0.06 mL, 0.1 mmol) were placed in a reaction vial. After stirring at 0° C. for 10 min, the reaction was diluted with acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product as a mixture of diastereomers. Example 21. C 38 H 41 F2N8O3(M+H) + LCMS calculated m / z = 695.3; found 695.3.

[0340] Example 22. 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Example 21, using 2-fluoroacrylic acid instead of (E)-4-methoxybut-2-enoic acid. Example 22. C 36 H 36 F3N8O2(M+H) + LCMS calculated m / z = 669.3; found 669.3.

[0341] Example 23a and Example 23b. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0342] Step 1. 3-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline [ka] A mixture of 4-bromo-3-methylisoquinoline (1.110 g, 5.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (2.285 g, 9.00 mmol), potassium acetate (1.472 g, 15.00 mmol), and PdCl2(dppf) (0.366 g, 0.500 mmol) in dioxane (20.0 mL) was stirred at 105 °C for 5 h. The solvent was removed and the product was purified by column eluted with hexane / EtOAc (up to 80% EtOAc). 16 H 21 BNO2(M+H) + LCMS calculated m / z = 270.1; found 270.1.

[0343] Step 2. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] tert-Butyl (2S,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (45.0 mg, 0.071 mmol), 3-methyl-4-(4,4 After evacuating and then refilling with N2 twice on a mixture of ,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline (28.8 mg, 0.107 mmol), potassium phosphate (45.4 mg, 0.214 mmol), and methanesulfonato(2-dicyclohexylphosphino-2',6'-dimethoxy-1,1'-biphenyl)(2'-methylamino-1,1'-biphenyl-2-yl)palladium(II) dichloromethane adduct (SPhos Pd G4) (5.68 mg, 7.14 μmol), the reaction was stirred at 95 °C for 4 h. The mixture was diluted with MeCN and then purified by preparative HPLC under PH = 10. The solvent was removed and the residue was dissolved in MeCN (1.0 mL), then TFA (1.0 mL) was added and the reaction was stirred at room temperature for 40 min. The solvent was removed and the crude product was used directly in the next step. C 34 H 38 FN8O (M+H) + LCMS calculated m / z = 593.3; found 593.4.

[0344] Step 3. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile To a solution of 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile (4.0 mg, 6.75 μmol), 2-fluoroacrylic acid (1.215 mg, 0.013 mmol), and 1-propanephosphonic acid cyclic anhydride (T3P 50% in EtOAc) (4.02 μl, 0.013 mmol) in ethyl acetate (0.8 ml) at 0° C., triethylamine (5.64 μl, 0.040 mmol) was added and the reaction was stirred for 30 min. The solvent was removed and the crude product was redissolved in acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the product as a TFA salt. Example 23a. Diastereomer 1. Peak 1.C 37 H 39 F2N8O2(M+H) + LCMS calculated m / z = 665.3; found 665.4. Example 23b. Diastereomer 2. Peak 2.C 37 H 39 F2N8O2(M+H) + LCMS calculated m / z = 665.3; found 665.4. 1 H NMR (600 MHz, DMSO) δ 9.46 (s, 1H), 8.32 (s, 1H), 8.27 (m, 1H), 7.72 (m, 2H), 7.22 (m, 1H), 5.94 (m 1H), 5.64 (m, 1H), 5.28-5.37 (m, 2H), 3.96 (q, 1H), 3.63 (m, 3H), 3.54 (m, 1H), 3.38 (m, 2H), 3.21 (m, 1H), 3.16 (s, 3H), 2.51(m, 4H), 2.40 (s, 3H), 2.34 (m, 1H), 2.19 (s, 3H), 2.13 (m, 1H), 1.97 (m, 2H), 1.59 (d, 3H).

[0345] Example 24a and Example 24b. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedures described in Step 3 of Example 23a and Example 23b using (E)-4-methoxybut-2-enoic acid instead of 2-fluoroacrylic acid. Example 24a. Diastereomer 1. Peak 1.C 39 H 44 FN8O3(M+H) + LCMS calculated m / z = 691.3; found 691.5. Example 24b. Diastereomer 2. Peak 2.C 39 H 44 FN8O3(M+H) + LCMS calculated m / z = 691.3; found 691.5.

[0346] Example 25a and Example 25b. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0347] Step 1. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Step 2 of Example 23a and Example 23b, using (7-fluoro-2-methylquinolin-8-yl)boronic acid instead of 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline. 34 H 37 F2N8O (M+H) + LCMS calculated m / z = 611.3; found 611.4.

[0348] Step 2. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile This compound was prepared according to the procedure described in step 3 of Example 23a and Example 23b, using 2-((2S,4S)-4-(6-fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile instead of 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile. Example 25a. Diastereomer 1. Peak 1.C 37 H 38 F3N8O2(M+H) + LCMS calculated m / z = 683.3; found 683.4. Example 25b. Diastereomer 2. Peak 2.C 37 H 38 F3N8O2(M+H) + LCMS calculated m / z = 683.3; found 683.4.

[0349] Example 26a and Example 26b. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0350] Step 1. (1-Methylisoquinolin-4-yl)boronic Acid [ka] A mixture of 4-bromo-1-methylisoquinoline (0.222 g, 1.0 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (0.457 g, 1.800 mmol), potassium acetate (0.294 g, 3.00 mmol), and PdCl2(dppf) (0.073 g, 0.100 mmol) in dioxane (5.0 ml) was stirred at 105° C. for 4 h. The product was purified by preparative HPLC under PH=2 (TFA). 10 H 11 BNO2(M+H) + LCMS calculated m / z = 188.1; found 188.1.

[0351] Step 2. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Step 2 of Example 23a and Example 23b, using (1-methylisoquinolin-4-yl)boronic acid instead of 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline. 34 H 38 FN8O (M+H) + LCMS calculated m / z = 593.3; found 593.4.

[0352] Step 3. 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile To a solution of 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile (4.0 mg, 6.75 μmol), (E)-4-methoxybut-2-enoic acid (1.567 mg, 0.013 mmol), and 1-propanephosphonic acid cyclic anhydride (T3P 50% in EtOAc) (4.02 μl, 0.013 mmol) in ethyl acetate (0.8 ml) at 0° C., triethylamine (5.64 μl, 0.040 mmol) was added and the reaction was stirred for 30 minutes. The solvent was removed and the crude product was redissolved in acetonitrile and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the product as a TFA salt. Example 26a. Diastereomer 1. Peak 1.C 39 H 44 FN8O3(M+H) + LCMS calculated m / z = 691.4; found 691.5. Example 26b. Diastereomer 2. Peak 2.C 39 H 44 FN8O3(M+H) + LCMS calculated m / z = 691.4; found 691.5.

[0353] Example 27a and Example 27b. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0354] Step 1. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Step 2 of Example 23a and Example 23b, using (7-fluoroquinolin-8-yl)boronic acid instead of 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline. 33 H 35 F2N8O (M+H) + LCMS calculated m / z = 597.3; found 597.4.

[0355] Step 2. 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile This compound was prepared according to the procedure described in step 3 of Example 23a and Example 23b, using 2-((2S,4S)-4-(6-fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile instead of 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile. Example 27a. Diastereomer 1. Peak 1.C 36 H36 F3N8O2(M+H) + LCMS calculated m / z = 669.3; found 669.4. Example 27b. Diastereomer 2. Peak 2. C 36 H 36 F3N8O2(M+H) + LCMS calculated m / z = 669.3; found 669.4.

[0356] Example 28a and Example 28b. 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indazol-3-yl)acetonitrile [ka]

[0357] Step 1. (4-Bromo-1-methyl-1H-indazol-3-yl)methanol [ka] To a mixture of methyl 4-bromo-1H-indazole-3-carboxylate (1.2 g, 4.70 mmol) and iodomethane (1.07 g, 7.5 mmol) in THF (20 mL) was added sodium hydride (0.34 g, 8.47 mmol, 60% dispersion in mineral oil) at 0° C. The reaction mixture was stirred for 5 min and allowed to warm to room temperature, at which point it was stirred overnight. Upon completion, the reaction mixture was quenched with aqueous NH4Cl (5 mL). The aqueous phase was extracted with EtOAc (10 mL×3), dried over MgSO4, filtered, and concentrated. The resulting crude mixture was dissolved in THF (20 mL) and cooled to −78° C., at which point LiAlH4 (11.7 mL, 11.7 mmol, 1 M in THF) was added slowly via syringe. The reaction mixture was warmed to 0° C. and stirred for an additional 15 min, at which point aqueous NH4Cl (10 mL) was added slowly. The aqueous phase was extracted with EtOAc (10 mL×3), dried over MgSO4, filtered, and concentrated. The crude mixture was used in the next step without further purification. C9H 10 BrNO (M+H) + LCMS calculated m / z = 241.0; found 241.0.

[0358] Step 2. 2-(4-Bromo-1-methyl-1H-indazol-3-yl)acetonitrile [ka] To (4-bromo-1-methyl-1H-indazol-3-yl)methanol (0.67 g, 2.77 mmol) in CH2Cl2 (10 mL) cooled at -78 °C, PBr3 (0.39 mL, 4.16 mmol) was added slowly. The reaction mixture was stirred at the same temperature for 1 h, at which point the reaction mixture was carefully quenched with aqueous NaHCO3 (5 mL). The aqueous phase was extracted with EtOAc (20 mL x 3), dried over MgSO4, filtered, and concentrated. DMSO (8 mL) was added to the crude mixture, followed by NaCN (0.41 g, 8.32 mmol) at room temperature. The reaction mixture was stirred at 60 °C for 3 h, and upon completion, cooled back to room temperature. Water (20 mL) was added to the reaction flask to precipitate the desired product. The desired product was then collected by filtration, washed with water, and air-dried for 4 h. C 10 H9BrN3(M+H) + LCMS calculated m / z = 250.0; found 250.0.

[0359] Step 3. 2-(1-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazol-3-yl)acetonitrile [ka] 2-(4-Bromo-1-methyl-1H-indazol-3-yl)acetonitrile (0.12 g, 0.48 mmol), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi(1,3,2-dioxaborolane) (0.18 g, 0.72 mmol), potassium acetate (0.15 g, 1.54 mmol), and PdCl2(dppf) (0.024 g, 0.033 mmol) in dioxane (3 mL) were stirred at 95 °C for 3 h. The solvent was removed and the product was purified by silica gel column eluted with hexane / EtOAc (up to 50% EtOAc). 16 H 21 BN3O2(M+H) + LCMS calculated m / z = 298.2; found 298.2.

[0360] Step 4. tert-Butyl (2R,4S)-4-(7-(3-(cyanomethyl)-1-methyl-1H-indazol-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate [ka] tert-Butyl (2R,4S)-4-(7-bromo-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate (40 mg, from step 3 of Example 14a and Example 14b) in dioxane / HO=4:1 (2 mL), To a solution of K3PO4 (49 mg, 0.23 mmol), 2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazol-3-yl)acetonitrile (29 mg, 0.099 mmol), and SPhosPdG4 (7.9 mg, 0.01 mmol) were added and the reaction mixture was heated to 90 °C for 1.5 h. Upon completion, the reaction was diluted with water, extracted with EtOAc (2 mL x 3), dried over MgSO4, filtered, and concentrated. The crude mixture was purified on silica gel eluting with CHCl2 / MeOH (up to 10% MeOH) to give the desired product as a mixture of diastereomers (20 mg, 44%). C 38 H 47 FN9O3(M+H) + LCMS calculated for: m / z = 696.4; found 696.4.

[0361] Step 5. 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indazol-3-yl)acetonitrile [ka] This compound is tert-butyl (2R,4S)-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate instead of tert-butyl (2R,4S)-4-( Prepared according to step 5 of Example 14a and Example 14b using 7-(3-(cyanomethyl)-1-methyl-1H-indazol-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate. The mixture was purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product. Example 28a. Diastereomer 1. Peak 1.C 36 H 41 FN9O2(M+H) + LCMS calculated m / z = 650.3; found 650.3. Example 28b. Diastereomer 2. Peak 2.C 36 H 41 FN9O2(M+H) + LCMS calculated m / z = 650.3; found 650.3.

[0362] Example 29. 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0363] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] After evacuating and refilling twice with N2 a mixture of intermediate 8 (2 g, 3.64 mmol), 6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (1.967 g, 5.46 mmol), potassium phosphate tribasic (3.86 g, 18.20 mmol), and Pd(Ph3P)4 (0.421 g, 0.364 mmol) in dioxane (25 ml) / water (12 ml), the reaction was stirred at 102° C. for 2 h. The mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc, and the combined organic layers were washed with brine, dried over Na2SO4, filtered, and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH2Cl2) to give the desired product as a brown solid. 36 H 41 F2N8O3S (M+H) + LCMS calculated m / z = 703.3; found 703.3.

[0364] Step 2. 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] To a solution of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (2.4 g, 3.41 mmol) in CH2Cl2 (30 ml) at 0° C. was added mCPBA (77% wet) (0.842 g, 3.76 mmol) and the reaction was stirred for 20 min at this temperature. The reaction was quenched by the addition of a saturated aqueous solution of Na2S2O3 and Na2CO3, diluted with CH2Cl2 and separated. The aqueous layer was extracted with CH2Cl2. The combined organic layers were dried over Na2SO4 and concentrated to give a crude mixture of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate and the corresponding sulfonyl compound. This crude material was used directly in the next step.

[0365] tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(6-fluoro-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (and corresponding sulfonyl compound, 2.4 g, 3.34 mmol) and N-ethyl-N,3-dimethylazetidin-3-amine (0.428 g, 3.34 mmol) were mixed and acetonitrile (33.4 ml) was added. After adding DIPEA (1.166 ml, 6.68 mmol) to the suspension, the mixture was heated to 70° C. and stirred at the same temperature for 2 hours. The solvent was evaporated to give the crude product as a brown solid.

[0366] tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate was dissolved in 10 ml of TFA, heated to 60° C. and stirred at the same temperature for 10 min. It was then cooled to room temperature, diluted with acetonitrile, filtered and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to separate the diastereomers (white amorphous powder, combined yield 53%). Diastereomer 1. Peak 1.C 32 H 37 F2N 10 (M+H) + LCMS calculated m / z = 599.3; found 599.3. Diastereomer 2. Peak 2.C 32 H 37 F2N 10 (M+H) + LCMS calculated m / z = 599.3; found 599.3.

[0367] Step 3. 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile bis(2,2,2-trifluoroacetate) (diastereomer 2, 395 mg, 0.477 mmol) was mixed with 2-fluoroacrylic acid (134 mg, 1.488 mmol) and then acetonitrile (10 mL) was added followed by DIPEA (780 μl, 4.46 mmol) and T3P (877 μl, 50% in AcOEt, 1.488 mmol). After stirring at room temperature overnight, the mixture was diluted with TFA, filtered and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give Example 29 (single diastereomer) as a TFA salt in the form of a white amorphous powder. Example 29. C 35 H 38 F3N 10 O (M+H) + LCMS calculated m / z = 671.3; found 671.3. 1H NMR (600 MHz, DMSO) δ 13.28 (s, br, 1H), 10.16 (s, br, 1H), 8.12 (s, 1H), 7.47-7.44 (m, 2H), 5.86 (td, J = 11.4, 5.5 Hz, 1H), 5.45 - 5.38 (m, 2H), 5.16 (m, 1H), 4.77-4.73 (m, 2H), 4.50-4.36 (m, 4H), 3.66-3.44 (m, 2H), 3.36 (m, 2H), 3.11 (m, 1H), 2.81 (s, 3H), 2.49-2.45 (m, 2H), 2.17 (s, 3H), 2.03 (s, 3H), 1.71 (s, 3H), 1.27 (t, J = 7.3 Hz, 3H).

[0368] Example 30. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0369] Step 1. tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] After evacuating and refilling twice with N2 a mixture of intermediate 8 (2 g, 3.64 mmol), 5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (1.945 g, 5.46 mmol), potassium phosphate tribasic (3.86 g, 18.20 mmol), and Pd(Ph3P)4 (0.421 g, 0.364 mmol) in dioxane (25 ml) / water (12 ml), the reaction was stirred at 102° C. for 2 h. The mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc, and the combined organic layers were washed with brine, dried over Na2SO4, filtered, and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH2Cl2) to give the desired product as a brown solid. 37 H 44 FN8O3S (M+H) + LCMS calculated m / z = 699.3; found 699.3.

[0370] Step 2. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] To a solution of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (2 g, 2.86 mmol) in CHCl (30 ml) at 0° C. was added mCPBA (0.962 g, 77% wet, 4.29 mmol) and the reaction was stirred at this temperature for 20 min. The reaction was quenched by the addition of a saturated aqueous solution of NaSO and NaCO, diluted with CHCl and separated. The aqueous layer was extracted with CHCl. The combined organic layers were dried over Na2SO4 and concentrated to give a crude mixture of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate and the corresponding sulfonyl compound. This crude material was used directly in the next step.

[0371] tert-Butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-8-methyl-4-(methylsulfinyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate (and the corresponding sulfonyl compound, 1.5 g, 2.098 mmol), N-ethyl-N,3-dimethylazetidin-3-amine (0.323 g, 2.52 mmol) were mixed and acetonitrile (20 ml) was added. After adding DIPEA (1.832 ml, 10.49 mmol) to the suspension, the mixture was heated to 70° C. and stirred at the same temperature for 2 hours. The solvent was evaporated to give the crude product as a brown solid.

[0372] The tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate obtained as above was dissolved in 10 ml of TFA, heated to 60° C. and stirred at the same temperature for 10 min. It was then cooled to room temperature, diluted with acetonitrile, filtered and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to separate the diastereomers (white amorphous powder, combined yield 51%). Diastereomer 1. Peak 1.C 33 H 40 FN 10 (M+H) + LCMS calculated m / z = 595.3; found 595.3. Diastereomer 2. Peak 2.C 33 H 40 FN 10 (M+H) + LCMS calculated m / z = 595.3; found 595.3.

[0373] Step 3. 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile bis(2,2,2-trifluoroacetate) (diastereomer 2, 400 mg, 0.486 mmol) and 2-fluoroacrylic acid (88 mg, 0.972 mmol) were mixed, followed by the addition of acetonitrile (10 mL) followed by the addition of DIPEA (849 μl, 4.86 mmol) and T3P (573 μl, 50% in AcOEt, 0.972 mmol). After stirring at room temperature overnight, the mixture was diluted with TFA, filtered and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give Example 30 (single diastereomer) as a TFA salt in the form of a white amorphous powder. Example 30.C 36 H 41 F2N 10 O (M+H) + LCMS calculated m / z = 667.3; found 667.3. 1 H NMR (600 MHz, DMSO) δ 13.02 (s, br, 1H), 10.18 (s, br, 1H), 8.10 (s, 1H), 7.47 (s, 1H), 7.31 (d, J = 5.1 Hz, 1H), 5.86 (td, J = 11.4, 5.5 Hz, 1H), 5.41 (dd, J = 18.2, 4.4 Hz, 1H), 5.38-4.62 (m, 9H), 3.73-3.56 (m, 2H), 3.36 (dd, J = 16.9, 6.8 Hz, 1H), 3.07 (td, J = 14.0, 5.2 Hz, 1H), 2.81 (s, 3H), 2.49 (s, 3H), 2.48-2.45 (m, 2H), 2.14 (s, 3H), 2.02 (s, 3H), 1.71 (s, 3H), 1.27 (t, J = 7.3 Hz, 3H).

[0374] Example 31a and Example 31b. 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile [ka]

[0375] Step 1. 2-(1-Methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)acetonitrile [ka] This compound was prepared according to step 3 of Example 28a and Example 28b, using 2-(4-bromo-1-methyl-1H-indazol-3-yl)acetonitrile instead of 2-(4-bromo-1-methyl-1H-indol-3-yl)acetonitrile. The mixture was purified using silica gel column chromatography eluting with hexane / EtOAc (up to 50% EtOAc). 17 H 22 BN2O2(M+H) + LCMS calculated m / z = 297.2; found 297.2.

[0376] Step 2. tert-Butyl (2R,4S)-4-(7-(3-(cyanomethyl)-1-methyl-1H-indol-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate [ka] This compound was prepared according to step 4 of Example 28a and Example 28b, using 2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)acetonitrile instead of 2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazol-3-yl)acetonitrile. The crude mixture was purified on silica gel eluting with CHCl / MeOH (up to 10% MeOH) to give the desired product as a mixture of diastereomers. 39 H 48 FN8O3(M+H) + LCMS calculated for: m / z = 695.4; found 695.4.

[0377] Step 3. 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile [ka] This compound is tert-butyl (2R,4S)-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate instead of tert-butyl (2R,4S)-4-( Prepared according to step 5 of Example 14a and Example 14b using 7-(3-(cyanomethyl)-1-methyl-1H-indol-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-methylpiperidine-1-carboxylate. The mixture was purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product. Example 31a. Diastereomer 1. Peak 1.C 37 H 42 FN8O2(M+H) + LCMS calculated m / z = 649.3; found 649.3. Example 31b. Diastereomer 2. Peak 2.C 37 H 42 FN8O2(M+H) + LCMS calculated m / z = 649.3; found 649.3. 1H NMR (600 MHz, DMSO) δ 8.17 (s, 1H), 7.62 (d, J = 8.3 Hz, 1H), 7.48 (s, 1H), 7.37 (dd, J = 8.3, 7.1 Hz, 1H), 7.00 (d, J = 7.1 Hz, 1H), 6.96-6.84 (m, 1H), 6.16 (dd, J = 16.6, 2.4 Hz, 1H), 5.95-5.85 (m, 1H), 5.73 (dd, J = 10.5, 2.4 Hz, 1H), 5.57 (dq, J = 12.4, 6.2 Hz, 1H), 4.73-4.62 (m, 1H), 4.31-4.20 (m, 1H), 3.97-3.91 (m, 1H), 3.88 (s, 3H), 3.65-3.54 (m, 2H), 3.20-3.07 (m, 6H), 2.51-2.45 (m, 2H), 2.38-2.25 (m, 2H), 2.24 (s, 3H), 2.15-1.90 (m, 4H), 1.57-1.43 (m, 6H).

[0378] Example 32a and Example 32b. 2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile [ka]

[0379] Step 1. tert-Butyl 4-(7-(3-(cyanomethyl)-1-methyl-1H-indol-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate [ka] This compound was prepared according to step 4 of Example 11a and Example 11b, using 2-(1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indol-3-yl)acetonitrile from step 1 of Example 31a and Example 31b instead of 8-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-naphthonitrile. The crude mixture was purified on silica gel eluting with CHCl / MeOH (up to 10% MeOH) to give the desired product as a mixture of diastereomers. 38 H 46 FN8O3(M+H) + LCMS calculated m / z = 681.4; found 681.4.

[0380] Step 2. 2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile [ka] This compound was prepared according to step 5 of Example 14a and Example 14b using tert-butyl 4-(7-(3-(cyanomethyl)-1-methyl-1H-indol-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2R,4S)-4-(7-(8-cyanonaphthalen-1-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate. The mixture was purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product. Example 32a. Diastereomer 1. Peak 1.C 36 H 40 FN8O2(M+H) + LCMS calculated m / z = 635.3; found 635.3. Example 32b. Diastereomer 2. Peak 2.C 36 H 40 FN8O2(M+H) + LCMS calculated m / z = 635.3; found 635.3. 1H NMR (600 MHz, DMSO) δ 8.26(s, 1H), 7.61 (d, J = 8.4 Hz, 1H), 7.47 (s, 1H), 7.45-7.35 (m, 1H), 7.00 (d, J = 7.1 Hz, 1H), 6.93 (dd, J = 16.7, 104 Hz, 1H), 6.18 (dd, J = 16.7, 2.5 Hz, 1H), 5.79-5.70 (m, 2H), 5.59-5.50 (m, 1H), 4.62-4.53 (m, 1H), 4.31-4.24 (m, 1H), 3.99-3.90 (m, 1H), 3.87 (s, 3H), 3.70-3.11 (m, 10H), 2.51-2.18 (m, 8H), 1.58 (d, J = 6.1 Hz, 3H).

[0381] Examples 33a and 33b. 2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka]

[0382] Step 1. 7-Bromo-2,4,6-trichloro-8-fluoro-3-nitroquinoline [ka] This compound was prepared following the procedure described in Intermediate 1, using NCS instead of NIS in step 1. C9H2BrCl3FN2O2 (M+H) + LCMS calculated m / z = 372.8; found 372.8.

[0383] Step 2. tert-Butyl (2S,4S)-4-(7-bromo-8-chloro-6-fluoro-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was prepared according to the procedure described in Step 1 of Intermediate 8, using 7-bromo-2,4,6-trichloro-8-fluoro-3-nitroquinoline instead of 7-bromo-2,4-dichloro-8-fluoro-6-iodo-3-nitroquinoline. 22 H 24 BrClFN6O2S (M+H) + LCMS calculated m / z = 569.1; found 569.1.

[0384] Step 3. tert-Butyl (2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-6-fluoro-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] tert-Butyl (2S,4S)-4-(7-bromo-8-chloro-6-fluoro-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (1 g, 1.755 mmol), 5,6-dimethyl-1-(tetrahydro-2H-pi) After evacuating and refilling with N2 twice for a mixture of (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (0.938 g, 2.63 mmol), tripotassium phosphate (1.862 g, 8.77 mmol), and Pd(Ph3P)4 (0.203 g, 0.175 mmol), the reaction was stirred at 102 °C for 2 h. The mixture was then cooled to room temperature, diluted with AcOEt and water, and separated. The aqueous layer was extracted with EtOAc, and the combined organic layers were washed with brine, dried over Na2SO4, filtered, and evaporated. The residue was purified by column chromatography (10-40% EtOAc in CH2Cl2) to give the desired product as a brown solid. C 36 H 41 ClFN8O3S (M+H) + LCMS calculated m / z = 719.3; found 719.3.

[0385] Step 4. tert-Butyl (2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate [ka] This compound was synthesized by the method of the present invention by using tert-butyl (2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl))-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate instead of tert-butyl (2S,4S)-2-(cyanomethyl)-4-(7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidine-1-carboxylate in step 2. Prepared according to the procedure described in steps 2-3 of example 6 using N-ethyl-N,3-dimethylazetidin-3-amine instead of N,N,3-trimethylazetidin-3-amine in step 3, using N-ethyl-N,3-dimethylazetidin-3-amine)-1H-indazol-4-yl)-6-fluoro-4-(methylthio)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate. The crude residue was purified by column chromatography (20-80% EtOAc in CH2Cl2) to give the desired product as a brown solid. C 42 H 53 ClFN 10 O3 (M+H) + LCMS calculated m / z = 799.4; found 799.4.

[0386] Step 5. 2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka] tert-Butyl (2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-2-(cyanomethyl)piperidine-1-carboxylate (427 mg, 0.534 mmol) was dissolved in TFA (5 mL). The mixture was stirred at 60° C. for 10 min, and then the solvent was removed.

[0387] The residue was mixed with 2-fluoroacrylic acid (144 mg, 1.601 mmol), followed by the addition of acetonitrile (10 mL), followed by the addition of DIPEA (0.93 mL, 5.34 mmol) and T3P (0.94 mL, 50% in AcOEt, 1.601 mmol). After stirring at room temperature overnight, the mixture was diluted with TFA, filtered and purified using preparative LCMS (XBridge C18 column, elution with a gradient of acetonitrile / water containing 0.1% TFA, flow rate 60 mL / min) to give the desired product as a TFA salt. Example 33a. Diastereomer 1. Peak 1.C 35 H 38 ClF2N 10 O (M+H) + LCMS calculated m / z = 687.3; found 687.3. Example 33b. Diastereomer 2. Peak 2.C 35 H 38 ClF2N 10 O (M+H) + LCMS calculated m / z = 687.3; found 687.3.

[0388] Example 34a and Example 34b. 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile [ka]

[0389] Step 1. 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Example 23a and Step 2 of Example 23b, using (2-fluoro-6-methoxyphenyl)boronic acid instead of 3-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)isoquinoline. 31 H 36 F2N7O2(M+H) + LCMS calculated m / z = 576.3; found 576.4.

[0390] Step 2. 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile This compound was prepared according to the procedure described in step 3 of Example 26a and Example 26b, using 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile instead of 2-((2S,4S)-4-(6-fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-2-yl)acetonitrile. Example 34a. Diastereomer 1. Peak 1.C 36 H 42 F2N7O4(M+H) + LCMS calculated m / z = 674.3; found 674.3. Example 34b. Diastereomer 2. Peak 2.C 36 H 42 F2N7O4(M+H) + LCMS calculated m / z = 674.3; found 674.3.

[0391] Examples 35a and 35b. 2-((2S,4S)-4-(8-chloro-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile [ka] This compound was prepared according to the procedure described in Example 33, using 6-fluoro-5-methyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole instead of 5,6-dimethyl-1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole in step 3. Example 35a. Diastereomer 1. Peak 1.C 34 H 35 ClF3N 10 O (M+H) + LCMS calculated m / z = 691.3; found 691.3. Example 35b. Diastereomer 2. Peak 2.C 34 H 35 ClF3N 10 O (M+H) + LCMS calculated m / z = 691.3; found 691.3.

[0392] Example A. GDP-GTP Exchange Assay. Inhibitor potency of the exemplified compounds was determined by a fluorescence-based guanine nucleotide exchange assay, which measures the exchange of Bodipy-GDP (fluorescently labeled GDP) for GppNHp (non-hydrolyzable GTP analog) to generate an active state of KRAS in the presence of SOS1 (a guanine nucleotide exchange factor). Inhibitors were serially diluted in DMSO and a volume of 0.1 μL was transferred to wells of a black low volume 384-well plate. A volume of 5 μL / well of Bodipy-loaded KRAS G12C diluted to 5 nM in assay buffer (25 mM Hepes pH 7.5, 50 mM NaCl, 10 mM MgCl2 and 0.01% Brij-35) was added to the plate and pre-incubated with the inhibitor for 2 hours at ambient temperature. Appropriate controls (enzyme without inhibitor or with G12C inhibitor (AMG-510)) were included on the plate. Exchange was initiated by the addition of a volume of 5 μL / well containing 1 mM GppNHp and 300 nM SOS1 in assay buffer. The 10 μL / well reaction concentrations of bodipy-loaded KRAS G12C, GppNHp, and SOS1 were 2.5 nM, 500 uM, and 150 nM, respectively. The reaction plate was incubated at ambient temperature for 2 hours, the time during which GDP-GTP exchange is expected to be complete in the absence of inhibitors. For KRAS G12D and G12V mutants, a similar guanine nucleotide exchange assay was used, with a final concentration of 2.5 nM for the bodipy-loaded KRAS protein and 4 and 3 hours of incubation after addition of the GppNHp-SOS1 mixture for G12D and G12V, respectively. A cyclic peptide described to selectively bind to the G12D mutant (Sakamoto et al., BBRC 484.3 (2017), 605-611) or an internal compound with confirmed binding was used as a positive control in the assay plate. Fluorescence intensity was measured with a PheraStar plate reader instrument (BMG Labtech) with excitation at 485 nm and emission at 520 nm.

[0393] Data were analyzed using either GraphPad Prism or Genedata Screener SmartFit. 50 Values ​​were derived by fitting the data to a four-parameter logistic equation that generates sigmoidal dose-response curve values ​​with variable Hill coefficients.

[0394] IC of KRAS_G12C exchange assay 50 Data, IC of KRAS_G12C pERK assay 50 Data, IC of KRAS_G12C WB pERK assay 50 The data are provided in Table 1 below. The symbol "†" indicates IC 50 ≤ 50 nM, "††" indicates IC 50 >50nM but ≤100nM, and "†††" indicates IC 50 is >100nM but ≤1000nM, and "††††" indicates IC 50 is >1 μM but ≦5 μM, and "†††††" indicates IC 50 >5 μM. "NA" indicates the absence of data. [Table 1-1] [Table 1-2]

[0395] Example B: Luminescent viability assay MIA PaCa-2 (KRAS G12C; ATCC® CRL-1420), NCI-H358 (KRAS G12C; ATCC® CRL-5807), A427 (KRAS G12D; ATCC® HTB53), HPAFII (KRAS G12D; ATCC® CRL-1997), YAPC (KRAS G12V; DSMZ ACC382), SW480 (KRAS G12V; ATCC® CRL-228), and NCI-H838 (KRAS WT; ATCC® CRL-5844) cells are cultured in RPMI 1640 medium supplemented with 10% FBS (Gibco / Life Technologies). 800 cells per well in RPMI 1640 medium supplemented with 2% FBS are seeded into white clear bottom 384-well Costar tissue culture plates containing a 50 nL dot of test compound (final concentration is 1:500 dilution in 0.2% DMSO). Plates are incubated at 370C, 5% CO2 for 3 days. 25 ul / well CellTiter-Glo reagent (Promega) is added at the end of the assay. Luminescence is read after 15 minutes using a PHERAstar (BMG). Data is analyzed in Genedata Screener using SmartFit to determine IC 50 Get the value.

[0396] Example C: Cellular pERK HTRF Assay MIA PaCa-2 (KRAS G12C; ATCC® CRL-1420), NCI-H358 (KRAS G12C; ATCC® CRL-5807), A427 (KRAS G12D; ATCC® HTB53), HPAFII (KRAS G12D; ATCC® CRL-1997), YAPC (KRAS G12V; DSMZ ACC382), SW480 (KRAS G12V; ATCC® CRL-228), and NCI-H838 (KRAS WT; ATCC® CRL-5844) cells were purchased from ATCC and maintained in RPMI 1640 medium supplemented with 10% FBS (Gibco / Life Technologies). Cells are plated at 5000 cells per well (8 μL) in Greiner 384-well low volume flat bottom tissue culture treated white plates and incubated overnight at 37°C, 5% CO2. The following morning, test compound stock solutions are diluted to 3x the final concentration in medium and 4 μL is added to the cells, resulting in a final DMSO concentration of 0.1%. Cells are incubated with test compounds for 4 hours (G12C and G12V) or 2 hours (G12D) at 37°C, 5% CO2. 4 μL of 4x lysis buffer containing blocking reagent (Cisbio) is added to each well and plates are rotated gently (300 rpm) for 30 minutes at room temperature. 4 μL per well of Cisbio anti-Phospho-ERK1 / 2d2 mixed with anti-Phospho-ERK1 / 2 cryptate (1:1) is added to each well and incubated overnight at room temperature in the dark. Plates are read on a Pherastar plate reader at wavelengths of 665 nm and 620 nm. Data is analyzed in Genedata Screener using SmartFit to determine IC 50 Get the value.

[0397] Example D: Whole Blood pERK1 / 2 HTRF Assay MIA PaCa-2 cells (KRAS G12C; ATCC® CRL-1420), HPAF-II (KRAS G12D; ATCC® CRL-1997), and YAPC (KRAS G12V; DSMZ ACC382) are maintained in RPMI 1640 with 10% FBS (Gibco / Life Technologies). For MIA PaCa-2 assays, cells are seeded in 96-well tissue culture plates (Corning, #3596) at 25000 cells per well in 100 μL of medium and cultured at 37° C., 5% CO2 for 2 days prior to the assay. For HPAF-II and YAPC assays, cells are seeded in 96-well tissue culture plates at 50000 cells per well in 100 uL of medium and cultured for 1 day prior to the assay. Whole blood is added to a 1 μL dot of compound (prepared in DMSO) in a 96-well plate and mixed gently by pipetting up and down so that the concentration of compound in the blood is 1x the desired concentration in 0.5% DMSO. Media is aspirated from the cells and 50 μL of whole blood is added per well containing the test compound and incubated at 37°C, 5% CO2 for 4 hours for MIA PaCa and YAPC assays or 2 hours for HPAF-II assays, respectively. After discarding the blood, the plate is gently washed twice by adding PBS to the side of the wells, discarding the PBS from the plate onto a paper towel, and tapping the plate thoroughly to drain. Then, 50 μL / well of 1x lysis buffer #1 (Cisbio) containing blocking reagent (Cisbio) and benzonase nuclease (Sigma catalog number E1014-5KU, final concentration 1:10000) is added and incubated at room temperature for 30 minutes with shaking (250 rpm). After lysis, 16 μL of lysate is transferred into a 384-well Greiner small volume white plate using Assist Plus (Integra Biosciences, NH). 4 μL of a 1:1 mixture of anti-Phospho-ERK1 / 2d2 and anti-Phospho-ERK1 / 2 cryptate (Cisbio) is added to the wells using Assist Plus and incubated overnight at room temperature in the dark.Plates are read on a Pherastar plate reader at wavelengths of 665 nm and 620 nm. Data is analyzed in Genedata Screener using SmartFit to determine ICs. 50 Get the value.

[0398] Example E: Ras Activation Elisa The 96-well Ras Activation ELISA Kit (Cell Biolabs Inc; #STA441) uses Raf1 RBD (Rho-binding domain) bound to a 96-well plate to selectively pull down the active form of Ras from cell lysates. The captured GTP-Ras is then detected by a pan-Ras antibody and an HRP-conjugated secondary antibody.

[0399] MIA PaCa-2 (KRAS G12C; ATCC® CRL-1420), NCI-H358 (KRAS G12C; ATCC® CRL-5807), A427 (KRAS G12D; ATCC® HTB53), HPAFII (KRAS G12D; ATCC® CRL-1997), YAPC (KRAS G12V; DSMZ ACC382), SW480 (KRAS G12V; ATCC® CRL-228), and NCI-H838 (KRAS WT; ATCC® CRL-5844) cells are maintained in RPMI 1640 containing 10% FBS (Gibco / Life Technologies). Cells are seeded in 96-well tissue culture plates (Corning, #3596) at 25000 cells per well in 100 μL of media and cultured at 37° C., 5% CO2 for 2 days so that they are approximately 80% confluent at the start of the assay. Cells are treated with compounds for either 4 hours or overnight at 37° C., 5% CO2. Upon harvesting, cells are washed with PBS, drained thoroughly, then lysed in 50 μL of 1x lysis buffer (provided by the kit) for 1 hour on ice with the addition of Halt Protease and Phosphatase Inhibitors (1:100).

[0400] Dilute Raf-1 RBD 1:500 in assay diluent (provided in the kit) and add 100 μL of diluted Raf-1 RBD to each well of the Raf-1 RBD capture plate. Cover the plate with plate sealing film and incubate on an orbital shaker for 1 hour at room temperature. Wash the plate 3 times with 250 μL of 1x wash buffer per well, aspirating thoroughly between each wash. Add 50 μL of Ras lysate sample (10-100 μg) in duplicate per well. Add "no cell lysate" controls in duplicate wells to determine background. Add 50 μL of assay diluent to all wells, each well immediately, and incubate the plate on an orbital shaker for 1 hour at room temperature. Wash the plate 5 times with 250 μL of 1x wash buffer per well, aspirating thoroughly between each wash. 100 μL of diluted anti-pan-Ras antibody is added to each well and the plate is incubated on an orbital shaker at room temperature for 1 hour. The plate is washed 5 times as above. 100 μL of diluted secondary antibody, HRP conjugate is added to each well and the plate is incubated on an orbital shaker at room temperature for 1 hour. The plate is washed 5 times as above and drained thoroughly. 100 μL of chemiluminescence reagent (provided in the kit) is added to each well, including blank wells. The plate is incubated on an orbital shaker at room temperature for 5 minutes before reading the luminescence of each microwell on a plate luminometer. % inhibition is calculated relative to DMSO control wells after subtracting the background level of the "no lysate control" from all values. IC 50 Determinations are made by curve fitting of percent inhibition versus log of inhibitor concentration using GraphPad Prism 7 software.

[0401] Example F: Inhibition of RAS-RAF and PI3K-AKT pathways The cellular potency of the compounds was determined by measuring the phosphorylation of KRAS downstream effectors extracellular signal-regulated kinase (ERK), ribosomal S6 kinase (RSK), AKT (also known as protein kinase B, PKB) and downstream substrate S6 ribosomal protein.

[0402] To measure phosphorylated extracellular signal-regulated kinase (ERK), ribosomal S6 kinase (RSK), AKT, and S6 ribosomal protein, cells (details regarding cell lines and types of data generated are detailed further in Table 2) were cultured at 4 × 10 4 Cells were seeded overnight in RPMI medium containing 200 cells / well. The next day, cells were incubated for 4 h at 37°C, 5% CO2 in the presence or absence of a range of test compounds. Cells were washed with PBS and lysed in 1x lysis buffer (Cisbio) containing protease and phosphatase inhibitors (Thermo Fisher, 78446). Ten or 20 μg of total protein lysates were subjected to SDS-PAGE and immunoblot analysis using the following antibodies: phospho-ERK1 / 2-Thr202 / Tyr204 (#9101L), total-ERK1 / 2 (#9102L), phosphor-AKT-Ser473 (#4060L), phospho-p90RSK-Ser380 (#11989S) and phospho-S6 ribosomal protein-Ser235 / Ser236 (#2211S) from Cell Signaling Technologies (Danvers, MA). [Table 2]

[0403] Example G: In vivo efficacy testing Mia-Paca-2 (KRAS G12C), H358 (KRAS G12C), HPAF-II (KRAS G12D), AGS (KRAS G12D), SW480 (KRAS G12V), or YAPC (KRAS G12V) human cancer cells are obtained from American Type Culture Collection and maintained in RPMI medium supplemented with 10% FBS. For efficacy study experiments, 5 × 10 6 Mia-Paca-2 cells are inoculated subcutaneously into the right hind flank of 6- to 8-week-old BALB / c nude mice (Charles River Laboratories, Wilmington, MA, USA). When the tumor volume is approximately 150-250 mm3, mice are randomized by tumor volume and administered compounds orally. Tumor volumes are calculated using the formula (L × W 2 ) / 2, where L and W refer to the length and width dimensions, respectively. Tumor growth inhibition was calculated using the formula (1-(V T / V C )) × 100, where V T is the tumor volume of the treatment group on the last day of treatment, and V C is the tumor volume of the control group on the last day of treatment. Two-way ANOVA with Dunnett's multiple comparison test is used to determine statistical differences between treatment groups (GraphPad Prism). Mice are housed with 10-12 animals per cage, fed with abundant food and exposed to a 12-h light / dark cycle. Mice with tumor volumes exceeding the limit (10% of body weight) are euthanized by CO2 inhalation. Animals are maintained in a barrier facility that is fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International. All procedures are performed in accordance with the US Public Service Policy on Human Care and Use of Laboratory Animals and with Incyte Animal Care and Use Committee Guidelines.

[0404] Example H: Caco2 Assay Caco-2 cells were grown at 37°C in a 5% CO2 atmosphere in DMEM growth medium supplemented with 10% (v / v) fetal bovine serum, 1% (v / v) non-essential amino acids, penicillin (100 U / mL), and streptomycin (100 μg / mL). Confluent cell monolayers of Caco-2 were subcultured every 7 or 4 days by treatment with 0.05% trypsin containing 1 μM EDTA. Caco-2 cells were seeded in 96-well Transwell plates. The seeding density of Caco-2 cells was 14,000 cells / well. After seeding, the DMEM growth medium was replaced every other day. For Caco-2 cells, cell monolayers were used for transport assays on days 22–25.

[0405] The cell culture medium was removed and replaced with HBSS. To measure TEER, HBSS was added to the donor compartment (apical) and receiver compartment (basolateral). TEER was measured by using a REMS autosampler to ensure the integrity of the cell monolayer. TEER values ​​≥ 300 Ω cm 2 Caco-2 cell monolayers with 100 μg / cm2 were used for transport experiments. app To determine the β-acetylglucosamine (β-glucosamine), a solution of the test compound in HBSS (50 μM) was added to the donor compartment (apical side) and a solution of 4% BSA in HBSS was added to the receiver compartment (basolateral). The apical volume was 0.075 mL and the basolateral volume was 0.25 mL. The incubation time was 120 min at 37° C. in a 5% CO2 atmosphere. At the end of the incubation time, samples were collected from the donor and receiver sides and proteins were precipitated by adding an equal volume of acetonitrile. After centrifugation (3000 rpm, Allegra X-14R Centrifuge supplied by Beckman Coulter, Indianapolis, IN), the supernatant was collected for LCMS analysis. The formula: P app (cm / s)=(F * VD) / (SA * MD) The permeation value was determined according to the following formula: where the flux rate (F, amount / time) was calculated from the slope of the cumulative amount of the target compound on the receiver side, SA is the surface area of ​​the cell membrane, VD is the donor volume, and MD is the initial amount of solution in the donor chamber.

[0406] Caco-2 permeability assay data is provided below in Table 3. The symbol "†" indicates Caco2 < 0.5, "††" indicates Caco-2 > 0.5 but < 1, and "†††" indicates Caco-2 > 1. "NA" indicates no Caco-2 data available. [Table 3]

[0407] Caco-2 permeability assay data for certain compounds from WO2021 / 142252 is provided in Table 4. [Table 4]

[0408] Example I: Human Whole Blood Stability Whole blood stability of example compounds was determined by LC-MS / MS. A 96-well Flexi-Tier™ Block (Analytical Sales & Services, Inc, Flanders, NJ) was used as an incubation plate containing 1.0 mL glass vials with 0.5 mL of blood (gender-mixed human whole blood sourced from BIOIVT, Hicksville, NY or similar source) per vial. Blood was pre-warmed to 37° C. in a water bath for 30 minutes. 96-deep-well analytical plates were prepared by adding 100 μL ultrapure water / well. 50 μL chilled ultrapure water / well was added to a 96-deep-well sample collection plate and covered with a sealing mat. 1 μL of 0.5 mM compound working solution (DMSO:water) was added to the blood in the incubation plate to a final concentration of 1 μM, mixed thoroughly by pipetting, and 50 μL was transferred to the T=0 well of the sample collection plate. After the blood is held in water for 2 minutes, 400 μL stop solution / well (acetonitrile with internal standard) is added. The incubation plate is placed in a 37°C Incu-Shaker CO2 Mini incubator (Benchmark Scientific, Sayreville, NJ) and shaken at 150 rpm. At 1, 2, and 4 hours, the blood samples are thoroughly mixed by pipetting and 50 μL is transferred to the corresponding wells of the sample collection plate. After the blood is held in water for 2 minutes, 400 μL stop solution / well is added. The collection plate is sealed and vortexed at 1700 rpm for 3 minutes (VX-2500 Multi-Tube Vortexer, VWR International, Radnor, PA), and the samples are centrifuged in the collection plate at 3500 rpm for 10 minutes (Allegra X-14R Centrifuge Beckman Coulter, Indianapolis, IN). Transfer 100 μL supernatant / well from the sample collection plate to the corresponding wells of the analysis plate. Vortex the final plate at 1700 rpm for 1 minute and analyze the samples by LC-MS / MS. The peak area ratios of the 1, 2, and 4 hour samples relative to T=0 are used to determine percent remaining.The natural logarithm of percent remaining versus time was used to determine the slope to calculate the compound half-life in blood (t 1 / 2 Calculate the slope (=0.693 / slope).

[0409] Human whole blood stability data is provided below in Table 5. The symbol "†" indicates WBS ≦70%, "††" indicates WBS >70% but ≦90%, and "†††" indicates WBS >90%. "NA" indicates no WBS data is available. [Table 5]

[0410] Example J: In Vitro Intrinsic Clearance Protocol For in vitro metabolic stability experiments, test compounds are incubated with human liver microsomes at 37°C. The incubation mixture contains test compound (1 μM), NADPH (2 mM), and human liver microsomes (0.5 mg protein / mL) in 100 mM phosphate buffer (pH 7.4). The mixture is pre-incubated at 37°C for 2 min before adding NADPH. The reaction is started upon addition of NADPH and quenched with ice-cold methanol at 0, 10, 20, and 30 min. The stopped incubation mixture is analyzed using an LC-MS / MS system. The analysis system consists of a Shimadzu LC-30AD binary pump system and a SIL-30AC autosampler (Shimadzu Scientific Instruments, Columbia, MD) coupled with a Sciex Triple Quad 6500+ mass spectrometer supplied by Applied Biosystems (Foster City, CA). Chromatographic separation of test compounds and internal standards is achieved using a Hypersil Gold C18 column (50×2.1 mm, 5 μM, 175 Å) supplied by ThermoFisher Scientific (Waltham, MA). Mobile phase A consists of 0.1% formic acid in water and mobile phase B consists of 0.1% formic acid in acetonitrile. The total run time of LC-MS / MS can be 2.75 min at a flow rate of 0.75 mL / min. Peak area integration and peak area ratio calculations are performed using Analyst software (version 1.6.3) supplied by Applied Biosystems.

[0411] In vitro intrinsic clearance (CL int,インビトロ ) is the t 1 / 2 From CL int,インビトロ =(0.693 / t 1 / 2 )×(1 / C タンパク質 ) where C タンパク質 is the protein concentration in the incubation, and t 1 / 2 is determined by the slope (k) of the log-linear regression analysis of the concentration versus time profile (hence, t1 / 2 Using physiologically based adjustment factors, liver microsomal protein concentration (45 mg protein / g liver), and liver weight (21 g / kg body weight), CL int,インビトロ Values ​​are scaled to in vivo values ​​for humans. CL int =CL int,インビトロ The formula is then used: × (mg protein / g liver weight) × (g liver weight / kg body weight). CL is then calculated in a well-mixed liver model that ignores all binding. int and hepatic blood flow Q (20 mL min in humans) -1 ·kg -1 ) to measure in vivo hepatic clearance (CL H ) to CL H =(Q × CL int ) / (Q+CL int ) CL divided by Q H The hepatic extraction rate was calculated as:

[0412] Example K: In vivo Pharmacokinetic Protocol For in vivo pharmacokinetic studies, test compounds are administered intravenously or by oral gavage to male Sprague-Dawley rats or male and female cynomolgus monkeys. For intravenous (IV) administration, test compounds are administered at 0.5-1 mg / kg via IV bolus for rats and via 5- or 10-min IV infusion for monkeys using a formulation of dimethylacetamide (DMAC) 10% in acidified saline. For oral (PO) administration, test compounds are administered at 1.0-3.0 mg / kg using methylcellulose 0.5% in citrate buffer (pH 2.5) containing 5% DMAC. Blood samples are collected pre-dose and at various time points up to 24 hours after dosing. All blood samples are collected using EDTA as an anticoagulant and centrifuged to obtain plasma samples. Plasma concentrations of test compounds are determined by LC-MS methods. Measured plasma concentrations are used to calculate PK parameters by standard non-compartmental methods using the Phoenix® WinNonlin software program (version 8.0, Pharsight Corporation).

[0413] Cassette dosing of test compounds is performed in rats and monkeys to obtain preliminary PK parameters.

[0414] In vivo pharmacokinetic studies using male beagle dogs may be carried out under the above conditions.

[0415] Example L: Time-Dependent Inhibition (TDI) Protocol for CYPs This assay is designed to characterize the increase in CYP inhibition associated with metabolism of the test compound over time. Potential mechanisms for this include the formation of tightly bound and quasi-irreversible inhibitory metabolite complexes or inactivation of P450 enzymes by covalent adduct formation of metabolites. Because 10-fold dilutions are used in this experiment to reduce metabolite concentrations, the effect of reversible inhibition is reduced, but it is possible (although not common) to obtain a positive result if the metabolite is a very potent CYP inhibitor.

[0416] Results are obtained using human liver microsomes (HLM) with a cocktail of CYP-specific probe substrates at 4x their Km concentrations for CYP2C9, 2C19, 2D6, and 3A4 (midazolam). HLM can be preincubated with test compounds at a concentration of 10 μM for 30 min in the presence (+N) or absence (-N) of an NADPH regenerating system, diluted 10-fold, and a new aliquot of NADPH regenerating system added and incubated for 8 min in the presence of the substrate cocktail. Using a calibration curve of metabolite standards, enzyme activity can be quantitatively measured using LC-MS / MS. Additionally, incubations containing known time-dependent inhibitors (thienilic acid (CYP2C9), ticlopidine (CYP2C19), paroxetine (CYP2D6), and troleandomycin (CYP3A4)) used as positive controls are preincubated for 30 min in the presence or absence of an NADPH regenerating system.

[0417] The analytical system consists of a Shimadzu LC-30AD binary pump system and a SIL-30AC autosampler (Shimadzu Scientific Instruments, Columbia, MD) coupled with a Sciex Triple Quad 6500+ mass spectrometer supplied by Applied Biosystems (Foster City, CA). Chromatographic separation of analytes and internal standards can be achieved using an ACQUITY UPLC BEH 130A, 2.1×50 mm, 1.7 μm HPLC column (Waters Corp, Milford, MA). Mobile phase A consists of water with 0.1% formic acid, and mobile phase B consists of acetonitrile with 0.1% formic acid. The total LC-MS / MS run time is 2.50 min at a flow rate of 0.9 mL / min. Peak area integration and peak area ratio calculations are performed using Analyst software (version 1.6.3) supplied by Applied Biosystems.

[0418] The percent of control CYP2C9, CYP2C19, CYP2D6, and CYP3A4 activity remaining after preincubation of compound with NADPH is calculated after correcting for the corresponding control vehicle activity, with 0 minutes set to 100%. A linear regression plot of the natural logarithm of % activity remaining versus time for each isoenzyme is used to calculate the slope. This slope represents the rate of enzyme decline, i.e., K obs is equivalent to.

[0419] Various modifications of the present invention in addition to those described herein will be apparent to those skilled in the art from the above description. Such modifications are also intended to fall within the scope of the appended claims. Each reference, including, but not limited to, all patents, patent applications, and publications cited in this application, is hereby incorporated by reference in its entirety.

Claims

1. Formula I: 【Chemical 1】 A compound having the formula or a pharmaceutically acceptable salt thereof, wherein Y is N or CH, R １ is selected from Cl, CH ３ , CH ２ F, CHF ２ , and CF ３ and is selected from Cy １ is 【Chemical Formula 2】 selected from R ２ is selected from F and Cl, R ３ is 【Chemical 3】 selected from Cy ２ is 【Chemical 4】 selected from provided that the compound of Formula I is 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile other than, a compound or a pharmaceutically acceptable salt thereof.

2. The compound of Formula I is a compound of Formula II: 【Chemical Formula 5】 or a pharmaceutically acceptable salt thereof, wherein R １ is selected from Cl and CH ３ and Cy １ is 【Chemical Formula 6】 selected from R ３ is 【Chemical Formula 7】 selected from Cy ２ is 【Chemical 8】 selected from, the compound according to claim 1 or a pharmaceutically acceptable salt thereof.

3. Y is N or CH, R １ is selected from Cl, CH ３ , CH ２ F, CHF ２ , and CF ３ and is selected from Cy １ is 【Chemical Formula 9】 selected from R ２ is selected from F and Cl, R ３ is 【Chemical 10】 selected from Cy ２ is 【Chemical 11】 selected from provided that the compound of Formula I is 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile The compound according to claim 1, or a pharmaceutically acceptable salt thereof, other than

4. Y is N or CH, R １ is selected from Cl, CH ３ , CH ２ F, CHF ２ , and CF ３ and is selected from Cy １ is 【Chemical 12】 selected from R ２ is selected from F and Cl, R ３ is 【Chemical 13】 selected from Cy ２ is 【Chemical Formula 14】 selected from the compound according to claim 1 or 3, or a pharmaceutically acceptable salt thereof.

5. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein Y is CH.

6. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, wherein Y is N.

7. Cy １ where Cy １ -a, Cy １ -b, Cy １ -c, Cy １ -d, and Cy １ -e, and the compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

8. Cy １ is Cy １ -f, Cy １ -g, Cy １ -h, Cy １ -i, Cy １ -j, and Cy １ -k, the compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

9. Cy １ is Cy １ -a and Cy １ -b, the compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

10. Cy １ is Cy １ -l, Cy １ -m, Cy １ -n, Cy １ -o, Cy １ -p, Cy １ -q, Cy １ -r, Cy １ -s, and Cy １ -t, the compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

11. Cy １ is Cy １ -c, Cy １ -m, Cy １ -n, Cy １ -o, Cy １ -p, Cy １ -q, Cy １ -r, Cy １ -s, and Cy １ selected from -t, the compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof.

12. R １ is CH ３ , CH ２ F, CHF ２ and CF ３ The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, selected from

13. R １ is CH ３ and CF ３ selected from, the compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

14. R ２ The compound according to claim 1 or 3, or a pharmaceutically acceptable salt thereof, wherein R is F.

15. R ２ The compound according to claim 1 or 3, or a pharmaceutically acceptable salt thereof, wherein R is Cl.

16. R ３ wherein R ３ is selected from R ３ -a and R -b, the compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

17. R ３ is R ３ -b and R ３ -c, the compound according to claim 1 or 3, or a pharmaceutically acceptable salt thereof.

18. Cy ２ is Cy ２ -a, Cy ２ -b, and Cy ２ -d, the compound according to claim 1 or 3, or a pharmaceutically acceptable salt thereof.

19. Cy ２ is Cy ２ -a and Cy ２ -b, a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

20. Cy ２ is Cy ２ -b and Cy ２ -d, a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof.

21. The compound of formula I is 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(2-methoxy-3-methylphenyl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(3-chloro-2-methoxyphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 1-(4-(6-Fluoro-7-(5-fluoroquinolin-8-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)piperidin-1-yl)prop-2-en-1-one, 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(2,3-dimethylphenyl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-8-methyl-7-(1-methyl-1H-indazol-6-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-8-methyl-7-(6-methylpyridin-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-8-methyl-7-(1-methyl-1H-indazol-3-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(4-fluorophenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 8-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-imidazo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(2-chloro-3-methylphenyl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-imidazo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 8-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)azetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 8-(6-fluoro-1-(1-((E)-4-fluorobut-2-enoyl)piperidin-4-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 8-(1-((2S,4S)-2-(cyanomethyl)-1-(2-fluoroacryloyl)piperidin-4-yl)-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-naphthonitrile, 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6,8-dichloro-4-(3-(dimethylamino)-3-methylazetidin-1-yl)-7-(5-fluoroquinolin-8-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-fluorobut-2-enoyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(4-(3-(dimethylamino)-3-methylazetidin-1-yl)-6-fluoro-7-(5-fluoroquinolin-8-yl)-8-(trifluoromethyl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile The compound according to claim 1 or 3, or a pharmaceutically acceptable salt thereof, selected from

22. The compound of formula I is 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(4-fluoroisoquinolin-1-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-8-methyl-7-(3-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-7-(7-fluoro-2-methylquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-8-methyl-7-(1-methylisoquinolin-4-yl)-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-((E)-4-methoxybut-2-enoyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-Fluoro-7-(7-fluoroquinolin-8-yl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indazol-3-yl)acetonitrile, 2-((2S,4S)-4-(4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-8-methyl-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-(4-(1-((2R,4S)-1-acryloyl-2-methylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile, 2-(4-(1-(1-acryloylpiperidin-4-yl)-6-fluoro-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-7-yl)-1-methyl-1H-indol-3-yl)acetonitrile, 2-((2S,4S)-4-(8-chloro-7-(5,6-dimethyl-1H-indazol-4-yl)-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, 2-((2S,4S)-4-(6-fluoro-7-(2-fluoro-6-methoxyphenyl)-8-methyl-4-((S)-1-((S)-1-methylpyrrolidin-2-yl)ethoxy)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile, and 2-((2S,4S)-4-(8-chloro-4-(3-(ethyl(methyl)amino)-3-methylazetidin-1-yl)-6-fluoro-7-(6-fluoro-5-methyl-1H-indazol-4-yl)-1H-[1,2,3]triazolo[4,5-c]quinolin-1-yl)-1-(2-fluoroacryloyl)piperidin-2-yl)acetonitrile The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from

23. A pharmaceutical composition comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier or additive.

24. A pharmaceutical for use in a method of inhibiting KRAS activity, comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, the method comprising contacting the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof with KRAS.

25. The pharmaceutical according to claim 24, wherein the contacting comprises administering the compound to a patient.

26. A pharmaceutical comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof for treating a disease or disorder associated with inhibition of KRAS interaction.

27. A pharmaceutical comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof for treating a disease or disorder associated with inhibition of a KRAS protein having a G12C mutation.

28. A pharmaceutical comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof for the treatment of cancer.

29. The pharmaceutical according to claim 28, wherein the cancer is selected from carcinoma, blood cancer, sarcoma, and glioblastoma.

30. The pharmaceutical according to claim 29, wherein the blood cancer is selected from myeloproliferative neoplasms, myelodysplastic syndromes, chronic and juvenile myelomonocytic leukemia, acute myeloid leukemia, acute lymphoblastic leukemia, and multiple myeloma.

31. The pharmaceutical according to claim 29, wherein the carcinoma is selected from pancreatic cancer, colorectal cancer, lung cancer, bladder cancer, gastric cancer, esophageal cancer, breast cancer, head and neck cancer, cervical cancer, skin cancer, and thyroid cancer.

32. The pharmaceutical according to claim 27, wherein the disease or disorder is an immune disorder or an inflammatory disorder.

33. The medicament according to claim 32, wherein the immune disorder or inflammatory disorder is a Ras-related lymphoproliferative disorder and juvenile myelomonocytic leukemia caused by a somatic mutation of KRAS.

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