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LIU HAIXIA (CN)
SHEN HONG (CN)
ZHANG WEIXING (CN)
ZHAO DAN (CN)
ZHU WEI (CN)
HOFFMANN LA ROCHE (US)
WO2023133543A1 | 2023-07-13 | |||
WO2023025832A1 | 2023-03-02 | |||
WO2022235870A1 | 2022-11-10 | |||
WO2022235864A1 | 2022-11-10 | |||
WO2023015559A1 | 2023-02-16 | |||
WO2022060836A1 | 2022-03-24 | |||
WO2021091982A1 | 2021-05-14 | |||
WO2021091967A1 | 2021-05-14 | |||
WO2021091956A1 | 2021-05-14 | |||
WO2022060836A1 | 2022-03-24 |
GENNARO, ALFONSO R ET AL.: "Remington: The Science and Practice of Pharmacy", 2000, LIPPINCOTT, WILLIAMS & WILKINS
ROWE, RAYMOND C: "Handbook of Pharmaceutical Excipients", 2005, PHARMACEUTICAL PRESS
CLAIMS 1. A compound of formula (I), , wherein R1 is 3-oxabicyclo[3.1.0]hexanyl, C1-6alkylC3-7cycloalkyl, cyanoC3-7cycloalkyl or haloC1- 6alkylC3-7cycloalkyl; R2 is H or halogen; R3 is H or halogen; R4 is C1-6alkyl or haloC1-6alkyl; R5 is C1-6alkoxyC1-6alkyl; R6 is morpholinyl, (haloC1-6alkyl)piperazinyl or C1-6alkylpiperazinyl; A1 is thiazolylene; A2 is C1-6alkylene; with the proviso that R2 and R3 are not H simultaneously; or a pharmaceutically acceptable salt thereof. 2. A compound of formula (Ia), , wherein R1 is 3-oxabicyclo[3.1.0]hexanyl, C1-6alkylC3-7cycloalkyl, cyanoC3-7cycloalkyl or haloC1- 6alkylC3-7cycloalkyl; R2 is H or halogen; R3 is H or halogen; R4 is C1-6alkyl or haloC1-6alkyl; R5 is C1-6alkoxyC1-6alkyl; R6 is morpholinyl, (haloC1-6alkyl)piperazinyl or C1-6alkylpiperazinyl; A1 is thiazolylene; A2 is C1-6alkylene; with the proviso that R2 and R3 are not H simultaneously; or a pharmaceutically acceptable salt thereof. 3. A compound according to claim 1 or 2, wherein R1 is C1-6alkylC3-7cycloalkyl or haloC1- 6alkylC3-7cycloalkyl. 4. A compound according to any one of claims 1-3, wherein R1 is methylcyclopropyl or (difluoromethyl)cyclopropyl. 5. A compound according to any one of claims 1-4, wherein R2 is H or fluoro. 6. A compound according to any one of claims 1-5, wherein R3 is H or fluoro. 7. A compound according to any one of claims 1-6, wherein R4 is ethyl or 2,2,2-trifluoroethyl. 8. A compound according to any one of claims 1-7, wherein R5 is 1-methoxyethyl. 9. A compound according to any one of claims 1-8, wherein R6 is morpholinyl or C1- 6alkylpiperazinyl. 10. A compound according to any one of claims 1-9, wherein R6 is morpholinyl or 4- methylpiperazin-1-yl. 11. A compound according to any one of claims 1-10, wherein A1 is , wherein bond “a” connects to indole ring. 12. A compound according to any one of claims 1-11, wherein A2 is dimethylmethylene. 13. A compound according to claim 1 or 2, wherein R1 is C1-6alkylC3-7cycloalkyl or haloC1-6alkylC3-7cycloalkyl; R2 is H or halogen; R3 is H or halogen; R4 is C1-6alkyl or haloC1-6alkyl; R5 is C1-6alkoxyC1-6alkyl; R6 is morpholinyl or C1-6alkylpiperazinyl; A1 is , wherein bond “a” connects to indole ring; A2 is C1-6alkylene; with the proviso that R2 and R3 are not H simultaneously; or a pharmaceutically acceptable salt thereof. 14. A compound according to claim 13, wherein R1 is 2-methylcyclopropyl or 2-(difluoromethyl)cyclopropyl; R2 is H or fluoro; R3 is H or fluoro; R4 is ethyl or 2,2,2-trifluoroethyl; R5 is (1S)-1-methoxyethyl; R6 is morpholinyl or 4-methylpiperazin-1-yl; A1 is , wherein bond “a” connects to indole ring; A2 is dimethylmethylene; with the proviso that R2 and R3 are not H simultaneously; or a pharmaceutically acceptable salt thereof. 15. A compound selected from: (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide; (1R,5S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- oxabicyclo[3.1.0]hexane-6-carboxamide; (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia- 2,5 9,13 22,26 9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen- 7-yl]cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin- 1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide; (1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]cyclopropanecarboxamide; (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino- 3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15- 2,5 9,13 22,26 oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropanecarboxamide; (1S,2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin- 1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide; (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]cyclopropanecarboxamide; and (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-morpholino-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide; or a pharmaceutically acceptable salt thereof. 16. A process for the preparation of a compound according to any one of claims 1 to 15 comprising any of the following steps: a) coupling reaction between compound of formula (II), the presence of a coupling reagent and a base to form the compound of formula (I); wherein R1, R2, R3, R4 , R5, R6, R7, A1 and A2 are defined as in any one of claims 1 to 14; the coupling reagent is T3P, HATU, PyBOP or EDCI/HOBt; the base is TEA, DIEPA or DMAP. 17. A compound or pharmaceutically acceptable salt according to any one of claims 1 to 15 for use as therapeutically active substance. 18. A pharmaceutical composition comprising a compound in accordance with any one of claims 1 to 15 and a pharmaceutically acceptable excipient. 19. The use of a compound according to any one of claims 1 to 15 for treating a KRAS G12C protein-related disease. 20. The use of a compound according to any one of claims 1 to 15 for treating a KRAS G12C, G12D and G12V protein-related disease. 21. The use of a compound according to any one of claims 1 to 15 for inhibiting RAS interaction with downstream effectors, wherein the downstream effectors are RAF and PI3K. 22. The use of a compound according to any one of claims 1 to 15 for inhibiting the propagating oncogenic MAPK and PI3K signaling. 23. The use of a compound according to any one of claims 1 to 15 for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic cancer, colorectal cancer, lung cancer, esophageal cancer, gallbladder cancer, melanoma ovarian cancer and endometrial cancer. 24. The use of a compound according to any one of claims 1 to 15 for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer. 25. A compound or pharmaceutically acceptable salt according to any one of claims 1 to 15 for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer. 26. The use of a compound according to any one of claims 1 to 15 for the preparation of a medicament for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer. 27. A method for the treatment or prophylaxis of KRAS mutation driven cancers, wherein the cancer is selected from pancreatic adenocarcinoma, colorectal cancer and non-small cell lung cancer, which method comprises administering a therapeutically effective amount of a compound as defined in any one of claims 1 to 15. 28. A compound or pharmaceutically acceptable salt according to any one of claims 1 to 15, when manufactured according to a process of claim 16. 29. The invention as hereinbefore described. |
Step 1: Preparation of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tert-butyldiphenylsil yl) oxy)-2,2-dimethylpropan-1-one (compound C3) To a mixture of 3-((tert-butyldiphenylsilyl)oxy)-2,2-dimethylpropanoyl chloride (compound C1, 35.0 g, 116.8 mmol) in DCM (400 mL) at 0 °C was added a solution of SnCl 4 (97.2 mL, 121.5 mmol) slowly. After being stirred at - 40 °C for 0.5 hour, 5-bromo-6-fluoro-1H- indole (compound C2, 25.0 g, 116.8 mmol) in DCM (200 mL) was added dropwise to the mixture which was stirred at - 40 °C for 15 min. After the reaction was completed, it was quenched with sat.NaHCO3 aq. (800 mL), and the reaction mixture was extracted with EtOAc (900 mL, twice). The combined organic layer was washed with brine (700 mL), dried over Na 2 SO 4 , filtered and concentrated in vacuo. The residue was triturated with the solution (100 mL, Petroleum ether: Ethyl acetate = 8:1) and filtered. The filter cake was dried in vacuo to afford 1- (5-bromo-6-fluoro-1H-indol-3-yl)-3-((tertbutyldiphenylsilyl) oxy)-2,2-dimethylpropan-1-one (compound C3, 50.0 g) as a yellow solid. MS calc’d 552.1 (MH + ), measured 552.1 (MH + ). Step 2: Preparation of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]- tert-butyl-diphenyl-silane (compound C4) To a mixture of 1-(5-bromo-6-fluoro-1H-indol-3-yl)-3-((tertbutyldiphenylsily l)oxy)-2,2- dimethylpropan-1-one (compound C3, 50.0 g, 90.49 mmol) in THF (600 mL) was added LiBH 4 (48.4 mL, 193.49 mmol, 4 M in THF) dropwise at 0 °C. The mixture was stirred at 70 °C for 24 hrs under nitrogen atmosphere. After the reaction was completed, it was quenched by addition of water (600 mL) at 0 °C slowly and the reaction mixture was extracted with EtOAc (600 mL, twice). The combined organic layer was washed with brine (600 mL), dried over Na 2 SO 4 , filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 20% ~ 33%) to afford [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl- propoxy]-tert-butyl-diphenyl-silane (compound C4, 46.0 g) as a white solid. MS calc’d 538.1 (MH + ), measured 538.2 (MH + ). Step 3: Preparation of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl- propoxy]-tert-butyl-diphenyl-silane (compound C5) To a mixture of [3-(5-bromo-6-fluoro-1H-indol-3-yl)-2,2-dimethyl-propoxy]-te rt-butyl- diphenyl-silane (compound C4, 35.4 g, 65.73 mmol) and iodine (18.4 g, 72.3 mmol) in THF (400 mL) was added silver trifluoromethanesulfonate (20.3 g, 78.88 mmol) at 0 °C. The mixture was stirred at 0 °C for 10 min. After the reaction was completed, it was quenched by sat. Na 2 SO 3 aq. (400 mL) and EtOAc (400 mL) and the reaction mixture was filtered. The organic layer was washed with brine (100 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 0% ~ 2.5%) to afford [3-(5- bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-t ert-butyl-diphenyl-silane (compound C5, 43.0 g) as a yellow solid. MS calc’d 664.0 (MH + ), measured 664.1 (MH + ). Step 4: Preparation of benzyl 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2- dimethyl-propyl]-6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyet hyl]-3-pyridyl]piperazine-1- carboxylate (compound C6) To a mixture of [3-(5-bromo-6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-prop oxy]-tert- butyl-diphenyl-silane (compound C5, 16.7 g, 25.13 mmol) and benzyl 4-[6-[(1S)-1- methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )-3-pyridyl]piperazine-1- carboxylate (Intermediate A, 16.7 g, 34.69 mmol) in a mixed solution of 1,4-dioxane (270 mL)/Toluene (90 mL) /water (90 mL) were added potassium phosphate (15.7 g, 73.92 mmol) and Pd(dppf)Cl2 (920 mg, 1.26 mmol). The mixture was stirred at 70 °C for 12 hrs under nitrogen atmosphere. After the reaction was completed, the mixture was filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 20% ~ 50%) to afford 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimeth yl-propyl]- 6-fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]pi perazine-1-carboxylate (compound C6, 19.5 g) as a white solid. MS calc’d 891.3 (MH + ), measured 891.3 (MH + ). Step 5: Preparation of benzyl 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy- 2,2-dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol- 2-yl]-6-[(1S)-1-methoxyethyl]-3- pyridyl]piperazine-1-carboxylate(compound C7) To a solution of 4-[5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2-dimeth yl-propyl]-6- fluoro-1H-indol-2-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]pipe razine-1-carboxylate (compound C6, 14.5 g, 16.26 mmol) and Cs 2 CO 3 (15.9 g, 48.77 mmol) in DMF (200 mL) was added 2,2,2- trifluoroethyl trifluoromethanesulfonate (37.7 g, 162.56 mmol) dropwise at 0 °C, and the mixture was stirred at 20 °C for 12 hrs. After the reaction was completed, EtOAc (70 mL) and water (100 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (70 mL, twice). Combined organic layer was washed with brine (100 mL, four times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica column chromatography to afford benzyl 4-[(5M)-5-[5-bromo-3-[3-[tert- butyl(diphenyl)silyl]oxy-2,2-dimethyl-propyl]-6-fluoro-1-(2, 2,2-trifluoroethyl)indol-2-yl]-6- [(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound C7, 8.0 g, PEAK 1, faster eluted) as yellow oil. MS calc’d 973.3 (MH + ), measured 973.2 (MH + ). Step 6: Preparation of benzyl 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2- dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S) -1-methoxyethyl]-3- pyridyl]piperazine-1-carboxylate (compound C8) To a solution of benzyl 4-[(5M)-5-[5-bromo-3-[3-[tert-butyl(diphenyl)silyl]oxy-2,2- dimethyl-propyl]-6-fluoro-1-(2,2,2-trifluoroethyl)indol-2-yl ]-6-[(1S)-1-methoxyethyl]-3- pyridyl]piperazine-1-carboxylate (compound C7, 10.5 g, 10.78 mmol) in DMF (130 mL) was added cesium fluoride (8.2 g, 53.9 mmol) and the mixture was stirred at 60 °C for 24 hrs. After the reaction was completed, EtOAc (100 mL) and water (100 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (100 mL, twice). The combined organic layer was washed with brine (80 mL, three times), dried over Na2SO4, filtered, and concentrated under vacuum to give a residue. The residue was purified by silica column chromatography (EtOAc in PE = 25% ~ 66%) to afford benzyl 4-[(5M)-5-[5-bromo-6-fluoro-3- (3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indo l-2-yl]-6-[(1S)-1-methoxyethyl]-3- pyridyl]piperazine-1-carboxylate (compound C8, 6.5 g) as a yellow solid. MS calc’d 735.2 (MH + ), measured 735.1 (MH + ). Step 7: Preparation of benzyl 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)- 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-tri fluoroethyl)indol-2-yl]-6-[(1S)-1- methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound C9) To a solution of benzyl 4-[(5M)-5-[5-bromo-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl )- 1-(2,2,2-trifluoroethyl)indol-2-yl]-6-[(1S)-1-methoxyethyl]- 3-pyridyl]piperazine-1-carboxylate (compound C8, 5.4 g) , bis(pinacolato)diboron (2.8 g, 11.01 mmol) and potassium acetate (1.2 mL, 18.35 mmol) in toluene (70 mL) was added Pd(dppf)Cl 2 (537.1 mg, 0.73 mmol). The mixture was degassed and purged with nitrogen atmosphere for three times and the mixture was stirred at 90 °C for 12 hrs. After the reaction was completed, the mixture was cooled to room temperature. The reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by silica column chromatography (EtOAc in PE = 25% ~ 66%) to afford benzyl 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4 ,5,5- tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl )indol-2-yl]-6-[(1S)-1- methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound C9, 5.2 g) as yellow oil. MS calc’d 783.3 (MH + ), measured 783.3 (MH + ). Step 8: Preparation of methyl (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4- benzyloxycarbonylpiperazin-1-yl)-2-[(1S)-1-methoxyethyl]-3-p yridyl]-6-fluoro-3-(3- hydroxy-2,2-dimethyl-propyl)-1-(2,2,2-trifluoroethyl)indol-5 -yl]thiazol-2-yl]-2-(tert- butoxycarbonylamino)-propanoyl]hexahydropyridazine-3-carboxy late (compound C10) To a mixture of methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ate (intermediate B, 2.7 g, 5.69 mmol), benzyl 4-[(5M)-5-[6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-5-(4,4 ,5,5- tetramethyl-1,3,2-dioxaborolan-2-yl)-1-(2,2,2-trifluoroethyl )indol-2-yl]-6-[(1S)-1- methoxyethyl]-3-pyridyl]piperazine-1-carboxylate (compound C9, 4.9 g, 6.32 mmol) in toluene (60 mL)/1,4-dioxane (20 mL) / water (20 mL) were added K 3 PO 4 (3.4 g, 15.81 mmol) and Pd(dtbpf)Cl 2 (412.2 mg, 0.63 mmol) under nitrogen atmosphere. The mixture was stirred at 70 °C for 12 hrs. After the reaction was completed, the mixture was concentrated in vacuo to give a residue. The residue was purified by silica column (EtOAc in PE = 10% ~ 75%) to afford methyl (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1 -yl)-2-[(1S)-1-methoxyethyl]-3- pyridyl]-6-fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-1-(2,2,2 -trifluoroethyl)indol-5-yl]thiazol-2- yl]-2-(tert-butoxycarbonylamino)-propanoyl]hexahydropyridazi ne-3-carboxylate (compound C10, 3.6 g) as a brown solid. MS calc’d 1053.4 (MH + ), measured 1053.3 (MH + ). Step 9: Preparation of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1 - yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy -2,2-dimethyl-propyl)-1-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbon ylamino)propanoyl]hexahy- dropyridazine-3-carboxylic acid (compound C11) To a solution of methyl (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1 - yl)-2-[(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy -2,2-dimethyl-propyl)-1-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbon ylamino)-propanoyl]- hexahydropyridazine-3-carboxylate (compound C10, 3.6 g, 3.42 mmol) in DCE (50 mL) was added trimethylstannanol (2.4 g, 13.67 mmol) and the mixture was stirred at 60 °C for 12 hrs. After the reaction was completed, EtOAc (80 mL) and water (60 mL) were added and the layers were separated. The aqueous phase was extracted with EtOAc (80 mL, twice). The combined organic layer was washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under vacuum to give (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1 -yl)-2- [(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-d imethyl-propyl)-1-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbon ylamino)propanoyl]hexahy- dropyridazine-3-carboxylic acid (compound C11, 4.3 g) as a brown solid. MS calc’d 1039.4 (MH + ), measured 1039.2 (MH + ). Step 10: Preparation of benzyl 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24- fluoro-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-1 5-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-(20M)- 20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carbox ylate (compound C12) To a mixture of (3S)-1-[(2S)-3-[4-[(2M)-2-[5-(4-benzyloxycarbonylpiperazin-1 -yl)-2- [(1S)-1-methoxyethyl]-3-pyridyl]-6-fluoro-3-(3-hydroxy-2,2-d imethyl-propyl)-1-(2,2,2- trifluoroethyl)indol-5-yl]thiazol-2-yl]-2-(tert-butoxycarbon ylamino)propanoyl]hexahy- dropyridazine-3-carboxylic acid (compound C11, 4.3 g, 4.14 mmol) in DCM (430 mL) was added DIEA (14.4 mL, 82.76 mmol), EDCI (11.9 g, 62.07 mmol) and 1-hydroxybenzotriazole (1.4 g, 10.35 mmol) at 0 °C. The mixture was stirred at 15 °C for 12 hrs. After the reaction was completed, the mixture was concentrated in vacuo, then diluted with water (80 mL), extracted with EtOAc (80 mL, twice). The combined organic layer was washed with brine (80 mL), dried over Na2SO4, filtered and concentrated in vacuo. The residue was purified by silica column chromatography (EtOAc in PE = 25% ~ 66%) to afford benzyl 4-[5-[(7S,13S)-7-(tert- butoxycarbonylamino)-24-fluoro-17,17-dimethyl-8,14-dioxo-21- (2,2,2-trifluoroethyl)-15-oxa-4- 2,5 9,13 22,26 thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23- hexaen-(20M)-20-yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]pipera zine-1-carboxylate (compound C12, 3.1 g) as yellow gum. MS calc’d 1021.4 (MH+), measured 1021.2 (MH+). Step 11: Preparation of tert-butyl N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-21-(2,2,2- 2,5 9,13 22,26 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (compound C13) To a mixture of benzyl 4-[5-[(7S,13S)-7-(tert-butoxycarbonylamino)-24-fluoro-17,17- dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia- 9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-(20M)-20- yl]-6-[(1S)-1-methoxyethyl]-3-pyridyl]piperazine-1-carboxyla te (compound C12, 3.1 g, 3.04 mmol) and formaldehyde aqueous (775.0 mg, 9.55 mmol) in methanol (150 mL) was added Pd(OH) 2 on activated carbon (2.79 g, 3.97 mmol). The mixture was degassed and purged with H 2 three times. The mixture was hydrogenated at 30 °C for 18 hrs. After the reaction was completed, the mixture was filtered and the filtrate was concentrated in vacuo to afford tert-butyl N- [(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-m ethylpiperazin-1-yl)-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamate (compound C13, 2.6 g) as a brown solid. MS calc’d 901.3 (MH + ), measured 901.3 (MH + ). Step 12: Preparation of (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-21-(2,2,2- 2,5 9,13 22,26 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (intermediate C) To a mixture of tert-butyl N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4 - methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-2 1-(2,2,2-trifluoroethyl)-15-oxa-4- 2,5 9,13 22,26 thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23- hexaen-7-yl]carbamate (compound C13, 2.6 g, 2.89 mmol) in DCM (18 mL) was added TFA (14.0 mL, 181.72 mmol). The mixture was stirred at 15 °C for 0.5 h. After the reaction was completed, the mixture was concentrated in vacuo and diluted with sat. NaHCO 3 (30 mL), extracted with EtOAc (30 mL, three times). The combined organic layer was washed with brine (50 mL), dried over Na2SO4, filtered and concentrated in vacuo to afford (7S,13S)-7-amino-24- fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazi n-1-yl)-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (intermediate C, 2.0 g) as a yellow solid, which was used directly in the next step. MS calc’d 801.3 (MH + ), measured 801.2 (MH + ) Intermediate D
(7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-m ethoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thi a-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14- dione The title compound was prepared in analogy to the preparation of Intermediate C by using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate. Intermediate E (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl] -5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21- (2,2,2-trifluoroethyl)-15-oxa-4- 2,5 9,13 22,26 thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23- hexaene-8,14-dione The compound was prepared according to the following scheme: E10 Intermediate E Step 1: Preparation of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2- trifluoroethyl)piperazine (compound E2). To a mixture of 3-bromo-5-iodo-2-[(1S)-1-methoxyethyl]pyridine (compound A3, 2.03 g, 5.95 mmol) and 1-(2,2,2-trifluoroethyl)piperazine (compound E1, 1.0 g, 5.95 mmol) in toluene (15 mL) were added Cs2CO3 (4.85 g, 14.88 mmol), (R)-binap (92.6 mg, 0.15 mmol) and Pd(OAc) 2 (66.8 mg, 0.3 mmol). The reaction mixture was degassed and purged with nitrogen for 3 times and the mixture was stirred at 100 °C for 12 hrs under nitrogen atmosphere. After being cooled to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography to 1-[5-bromo-6- [(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2-trifluoroethyl)pip erazine (compound E2, 2.0 g) as yellow oil. MS calc’d 382.2 (MH + ), measured 382.1 (MH + ) Step 2: 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-diox aborolan-2-yl)-3- pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (compound E3). To a solution of 1-[5-bromo-6-[(1S)-1-methoxyethyl]-3-pyridyl]-4-(2,2,2- trifluoroethyl)piperazine (compound E2, 3.2 g, 8.37 mmol), bis(pinacolato)diboron (3.19 g, 12.56 mmol) and KOAc (2.1 g, 20.93 mmol) in toluene (50 mL) was added Pd(dppf)Cl2 (306.3 mg, 0.42 mmol). The mixture was degassed and purged with nitrogen for 3 times and the mixture was stirred at 90 °C for 12 hrs under nitrogen atmosphere. After being cooled to the room temperature, the reaction mixture was filtered, the filtrate was concentrated in vacuo to give a residue, which was purified by reversed phase column to afford 1-[6-[(1S)-1-methoxyethyl]-5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-3-pyridyl]-4-( 2,2,2-trifluoroethyl)piperazine (compound E3, 1.9 g) as a yellow gum. MS calc’d 430.2 (MH + ), measured 348.4 (M-C6H10+H + ). Step 3: Preparation of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1H-indol-3-yl]-2,2 -dimethyl-propoxy]-tert-butyl- diphenyl-silane (compound E4). To a solution of 1-[6-[(1S)-1-methoxyethyl]-5-(4,4,5,5-tetramethyl-1,3,2-diox aborolan-2- yl)-3-pyridyl]-4-(2,2,2-trifluoroethyl)piperazine (compound E3, 1.9 g, 4.41 mmol), [3-(5-bromo- 6-fluoro-2-iodo-1H-indol-3-yl)-2,2-dimethyl-propoxy]-tert-bu tyl-diphenyl-silane (compound C5, 2.1 g, 3.15 mmol) in 1,4-dioxane (24 mL), water (8 mL) and toluene (8 mL) was added K3PO4 (2.1 g, 9.5 mmol) and Pd(dppf)Cl 2 (231 mg, 0.37 mmol). The mixture was degassed by bubbling nitrogen for 2 min, and the reaction mixture was stirred at 70 °C for 12 hrs. After being cooled to room temperature, the reaction mixture was filtered. The filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE : 30% - 60%) to afford [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin- 1-yl]-3-pyridyl]-1H-indol-3-yl]-2,2-dimethyl-propoxy]-tert-b utyl-diphenyl-silane (compound E4, 960.0 mg) as a yellow gum. MS calc’d 839.3 (MH + ), measured 839.3 (MH + ) Step 4: Preparation of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4- (2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-tr ifluoroethyl)indol-3-yl]-2,2- dimethyl-propoxy]-tert-butyl-diphenyl-silane (compound E5). To a solution of [3-[5-bromo-6-fluoro-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1H-indol-3-yl]-2,2 -dimethyl-propoxy]-tert-butyl- diphenyl-silane (compound E4, 1 g, 1.14 mmol) in DMF (35 mL) was added Cs2CO3 (1.1 g, 3.44 mmol) and 2,2,2-trifluoroethyl trifluoromethanesulfonate (2.7 g, 11.63 mmol) at 0 °C. After being stirred at 20 °C for 15 hrs, the reaction mixture was poured into water (100 mL), and extracted with EtOAc (50 mL, three times). The combined organic was washed with brine (50 mL, three times), dried over Na2SO4, filtered and concentrated under vacuum to give a residue which was purified by column chromatography (EtOAc in PE: 30% - 40%) to afford [3-[5- bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-t rifluoroethyl)piperazin-1-yl]-3- pyridyl]-1-(2,2,2-trifluoroethyl)indol-3-yl]-2,2-dimethyl-pr opoxy]-tert-butyl-diphenyl-silane (compound E5, 640.0 mg, faster eluted) as a white solid. MS calc’d 921.3 (MH + ), measured 921.4 (MH + ). Step 5: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4- (2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-tr ifluoroethyl)indol-3-yl]-2,2- dimethyl-propan-1-ol (compound E6). To a solution of [3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2 ,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoro ethyl)indol-3-yl]-2,2-dimethyl- propoxy]-tert-butyl-diphenyl-silane (compound E5, 640.0 mg, 0.69 mmol) in DMF (7 mL) was added cesium fluoride (421.8 mg, 2.78 mmol). The mixture was stirred at 60 °C for 16 hrs. After being cooled to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE : 30% - 60%) to afford 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5- [4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2 -trifluoroethyl)indol-3-yl]-2,2- dimethyl-propan-1-ol (compound E6, 360.0 mg) as yellow oil. MS calc’d 683.2 (MH + ), measured 683.1 (MH + ). Step 6: Preparation of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4- (2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-tr ifluoroethyl)indol-3-yl]-2,2- dimethyl-propan-1-ol (compound E7). To a solution of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2, 2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoro ethyl)indol-3-yl]-2,2-dimethyl-propan- 1-ol (compound E6, 360.0 mg, 0.53 mmol), bis(pinacolato)diboron (200.6 mg, 0.79 mmol) in toluene (6 mL) was added potassium acetate (0.08 mL, 1.32 mmol) and Pd(dppf)Cl2 (40 mg, 0.1 mmol). The reaction mixture was degassed by bubbling nitrogen for 5 min then stirred at 80 °C for 15 hrs. After being cooled to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE : 30% - 50%) to afford 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1- methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-p yridyl]-1-(2,2,2-trifluoroethyl)indol- 3-yl]-2,2-dimethyl-propan-1-ol (compound E7, 300.0 mg) as yellow gum. MS calc’d 731.4 (MH + ), measured 731.4 (MH + ). Step 7: Preparation of methyl (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6- fluoro-3-(3-hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-m ethoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoro ethyl)indol-5-yl]thiazol-2- yl]propanoyl]hexahydropyridazine-3-carboxylate (compound E8). To a mixture of 3-[5-bromo-6-fluoro-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2, 2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoro ethyl)indol-3-yl]-2,2-dimethyl-propan- 1-ol (compound E7, 0.3 g, 0.41 mmol) and methyl (3S)-1-[(2S)-3-(4-bromothiazol-2-yl)-2-(tert- butoxycarbonylamino)propanoyl]hexahydropyridazine-3-carboxyl ate (intermediate B, 196.7 mg, 0.41 mmol) in toluene (3 mL), 1,4-dioxane (1 mL) and water (1 mL) were added K3PO4 (221.3 mg, 1.04 mmol) and Pd(dtbpf)Cl 2 (27.05 mg, 0.04 mmol). The mixture was stirred at 70 °C for 12 hrs under nitrogen atmosphere. After being cooled to room temperature, the reaction mixture was filtered and the filtrate was concentrated in vacuo to give a residue. The residue was purified by column chromatography (EtOAc in PE : 60% - 80%) to afford methyl (3S)-1-[(2S)-2-(tert- butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethy l-propyl)-(2M)-2-[2-[(1S)-1- methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-p yridyl]-1-(2,2,2-trifluoroethyl)indol- 5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylat e (compound E8, 200.0 mg) as yellow gum. MS calc’d 1001.4 (MH + ), measured 1001.4 (MH + ). Step 8: Preparation of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-( 3- hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl] -5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoro ethyl)indol-5-yl]thiazol-2- yl]propanoyl]hexahydropyridazine-3-carboxylic acid (compound E9). To a mixture of methyl (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-( 3- hydroxy-2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl] -5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-1-(2,2,2-trifluoro ethyl)indol-5-yl]thiazol-2- yl]propanoyl]hexahydropyridazine-3-carboxylate (compound E8, 200.0 mg, 0.2 mmol) in DCE (5 mL) was added Me3SnOH (200.0 mg, 1.11 mmol). The mixture was stirred at 60 °C for 12 hrs. The reaction mixture was concentrated under vacuum to give a residue. EtOAc (10 mL) and water (10 mL) were added to the residue and the layers were separated. The aqueous phase was extracted with EtOAc (15 mL, twice). The combined organic layer was washed with brine (20 mL), dried over Na 2 SO 4 , filtered, and concentrated under vacuum to afford (3S)-1-[(2S)-2-(tert- butoxycarbonylamino)-3-[4-[6-fluoro-3-(3-hydroxy-2,2-dimethy l-propyl)-(2M)-2-[2-[(1S)-1- methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-p yridyl]-1-(2,2,2-trifluoroethyl)indol- 5-yl]thiazol-2-yl]propanoyl]hexahydropyridazine-3-carboxylic acid (compound E9, 188.0 mg) as a brown solid. MS calc’d 987.4 (MH + ), measured 987.4 (MH + ). Step 9: Preparation of tert-butyl N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-p yridyl]-17,17-dimethyl-8,14- dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamate (compound E10). To a mixture of (3S)-1-[(2S)-2-(tert-butoxycarbonylamino)-3-[4-[6-fluoro-3-( 3-hydroxy- 2,2-dimethyl-propyl)-(2M)-2-[2-[(1S)-1-methoxyethyl]-5-[4-(2 ,2,2-trifluoroethyl)piperazin-1- yl]-3-pyridyl]-1-(2,2,2-trifluoroethyl)indol-5-yl]thiazol-2- yl]propanoyl]hexahydropyridazine-3- carboxylic acid (compound E9, 188.0 mg, 0.19 mmol) in DCM (20 mL) were added DIEA (0.7 mL, 3.81 mmol), EDCI (550.0 mg, 2.87 mmol) and HOBt (65.0 mg, 0.48 mmol) at 0 °C. After being stirred at 20 °C for 12 hrs, the reaction mixture was poured into water (20 mL) and extracted with EtOAc (20 mL, three times). The combined organic layer was washed with brine (30 mL), dried over Na2SO4, filtered and concentrated under vacuum to give a residue which was purified by column chromatography (EtOAc in PE : 50% - 70%) to afford tert-butyl N-[(7S,13S)- 24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trif luoroethyl)piperazin-1-yl]-3- pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl) -15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]carbamate (compound E10, 110.0 mg) as a yellow solid. MS calc’d 969.4 (MH+), measured 969.5 (MH + ). Step 10: Preparation of (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piperazin-1-yl]-3-p yridyl]-17,17-dimethyl-21- 2,5 9,13 22,26 (2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapenta cyclo[17.5.2.1 .1 .0 ]- octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate E). To a solution of tert-butyl N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-[4 - (2,2,2-trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimet hyl-8,14-dioxo-21-(2,2,2- 2,5 9,13 22,26 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaen-7-yl]carbamate (compound E10, 110.0 mg, 0.11 mmol) in DCM (1 mL) was added TFA (1.0 mL, 12.98 mmol). The mixture was stirred at 20 °C for 1 h. After the reaction was completed, the reaction mixture was concentrated under vacuum to give a residue. Sat. NaHCO3 aq. (20 mL) was added and the mixture was extracted with EtOAc (15 mL, three times). The combined organic layer was washed with brine (20 mL), dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to afford (7S,13S)-7-amino-24-fluoro-(20M)- 20-[2-[(1S)-1-methoxyethyl]-5-[4-(2,2,2-trifluoroethyl)piper azin-1-yl]-3-pyridyl]-17,17- dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]-octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14- dione (Intermediate E, 98.0 mg) as a yellow solid. MS calc’d 869.4 (MH + ), measured 869.2 (MH + ). Intermediate F (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15- oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14- dione The title compound was prepared in analogy to the preparation of Intermediate E by using iodoethane instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate. Intermediate G (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-morpholino-3- pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapent acyclo- 2,5 9,13 22,26 [17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione The title compound was prepared in analogy to the preparation of Intermediate E by using by using iodoethane and morpholine instead of 2,2,2-trifluoroethyl trifluoromethanesulfonate and 1-(2,2,2-trifluoroethyl)piperazine (compound E1). Intermediate H (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl] -5-(4-methylpiperazin-1-yl)- 3-pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14- dione The title compound was prepared in analogy to the preparation of Intermediate C by using 5-bromo-4-fluoro-1H-indole instead of 5-bromo-6-fluoro-1H-indole (compound C2). Intermediate I (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl] -5-morpholino-3-pyridyl]- 17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21, 27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14- dione
The title compound was prepared in analogy to the preparation of Intermediate E by using by using morpholine instead of 1-(2,2,2-trifluoroethyl)piperazine (compound E1). Example 1 (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-m ethoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-1 5-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide To a solution of (1S,2S)-2-methylcyclopropanecarboxylic acid (compound 1a, 64.3 mg, 0.64 mmol) in DMF (2 mL) were added DIEA (0.3 mL, 1.61 mmol), HATU (366.5 mg, 0.96 mmol) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thi a-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (intermediate D, 240.0 mg, 0.32 mmol) at 0 °C. After being stirred at 20 °C for 16 hrs, the reaction mixture was poured into ice water (10 mL) and extracted with EA (20 mL, three times). The combined organic layer was washed with brine (20 mL), dried over Na2SO4 and concentrated under vacuum to give a residue. The resulting residue was purified by silica gel chromatography to afford (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-15-oxa-4-thia- 2,5 9,13 22,26 9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen- 7-yl]-2-methyl-cyclopropanecarboxamide (Example 1, 156.41 mg) as a white solid. MS calc’d 829.5 (MH + ), measured 829.5 (MH + ). 1 H NMR (400 MHz, METHANOL-d 4 ) δ = 8.64 (d, J = 7.6 Hz, 1H), 8.58 (d, J = 8.4 Hz, 1H), 8.42 (d, J = 2.9 Hz, 1H), 7.63 (d, J = 2.4 Hz, 1H), 7.34 (d, J = 2.8 Hz, 1H), 7.31 (d, J = 12.7 Hz, 1H), 5.79 - 5.73 (m, 1H), 4.44 (br d, J = 11.4 Hz, 1H), 4.28 - 4.13 (m, 4H), 3.77 - 3.68 (m, 2H), 3.41 - 3.37 (m, 4H), 3.28 - 3.13 (m, 2H), 3.02 (br d, J = 13.6 Hz, 1H), 2.77 - 2.74 (m, 4H), 2.63 (br d, J = 14.3 Hz, 1H), 2.44 (s, 3H), 2.21 - 2.13 (m, 1H), 1.97 (s, 2H), 1.68 - 1.58 (m, 1H), 1.51 - 1.47 (m, 1H), 1.42 (d, J = 6.2 Hz, 2H), 1.37 (d, J = 6.6 Hz, 4H), 1.13 (d, J = 6.0 Hz, 3H), 1.08 (br dd, J = 4.2, 8.6 Hz, 1H), 0.98 (t, J = 7.0 Hz, 3H), 0.95 - 0.89 (m, 4H), 0.67 - 0.62 (m, 1H), 0.50 (s, 3H) ppm. Example 2 (1R,5S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-m ethoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-1 5-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-3- oxabicyclo[3.1.0]hexane-6-carboxamide The title compound was prepared in analogy to the preparation of Example 1 by using (1S,5R)-3-oxabicyclo[3.1.0]hexane-6-carboxylic acid instead of (1S,2S)-2- methylcyclopropanecarboxylic acid (compound 1a). Example 2 (10.2 mg) was obtained as a white solid. MS calc’d 857.4 (MH + ), measured 857.1 (MH + ). 1 H NMR (400 MHz, ACETONITRILE-d 3 ) δ = 8.61 (d, J = 7.6 Hz, 1H), 8.41 (d, J = 2.8 Hz, 1H), 7.62 (d, J = 2.5 Hz, 1H), 7.33 - 7.28 (m, 1H), 7.18 (d, J = 2.8 Hz, 1H), 7.04 (br d, J = 9.5 Hz, 1H), 5.68 (br t, J = 8.8 Hz, 1H), 4.41 - 4.30 (m, 2H), 4.20 - 4.05 (m, 4H), 3.88 - 3.83 (m, 2H), 3.71 - 3.64 (m, 4H), 3.37 (br d, J = 14.9 Hz, 1H), 3.28 - 3.24 (m, 4H), 3.22 - 3.17 (m, 3H), 3.13 (br dd, J = 9.1, 15.0 Hz, 1H), 2.91 (br d, J = 14.9 Hz, 1H), 2.70 (dt, J = 2.7, 12.9 Hz, 1H), 2.58 (br d, J = 14.1 Hz, 1H), 2.50 (br t, J = 4.8 Hz, 4H), 2.30 - 2.22 (m, 4H), 1.78 - 1.72 (m, 1H), 1.57 - 1.50 (m, 2H), 1.36 (d, J = 6.1 Hz, 3H), 1.27 (br s, 1H), 0.96 (br t, J = 7.1 Hz, 3H), 0.88 (s, 3H), 0.47 (s, 3H) ppm. Example 3 (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(2 0M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-8,14-dioxo-15-oxa-4- 2,5 9,13 22,26 thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23- hexaen-7-yl]cyclopropanecarboxamide The title compound was prepared in analogy to the preparation of Example 1 by using (1S,2S)-2-(difluoromethyl)cyclopropanecarboxylic acid instead of (1S,2S)-2- methylcyclopropanecarboxylic acid (compound 1a). Example 3 (31.4 mg) was obtained as a yellow solid. MS calc’d 865.4 (MH + ), measured 865.1 (MH + ). 1 H NMR (400 MHz, CHLOROFORM-d) δ = 8.76 (br d, J = 2.6 Hz, 1H), 8.66 (d, J = 7.6 Hz, 1H), 7.63 (d, J = 2.1 Hz, 1H), 7.30 (br d, J = 2.4 Hz, 1H), 7.12 (d, J = 12.3 Hz, 1H), 6.78 (br d, J = 9.9 Hz, 1H), 6.00 - 5.92 (m, 1H), 4.66 - 4.53 (m, 1H), 4.36 (q, J = 6.2 Hz, 1H), 4.31 - 4.23 (m, 1H), 4.19 - 4.04 (m, 2H), 3.89 - 3.86 (m, 1H), 3.78 - 3.66 (m, 4H), 3.47 - 3.44 (m, 1H), 3.41 (s, 3H), 3.24 - 3.05 (m, 6H), 2.93 (s, 3H), 2.89 - 2.80 (m, 1H), 2.78 - 2.70 (m, 1H), 2.53 - 2.39 (m, 1H), 2.29 - 2.18 (m, 1H), 2.08 - 1.66 (m, 5H), 1.53 - 1.45 (m, 3H), 1.19 - 1.14 (m, 2H), 1.37 - 1.08 (m, 2H), 1.02 - 0.87 (m, 6H), 0.50 (s, 3H) ppm. Example 4 (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyeth yl]-5-(4-methylpiperazin-1- yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoro ethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl] -5-(4-methylpiperazin-1-yl)-3- pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-t hia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate C) instead of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D). Example 4 (15.7 mg) was obtained as a white solid. MS calc’d 883.5 (MH + ), measured 883.1 (MH + ). 1 H NMR (400MHz, Methanol-d4) δ = 8.67 (d, J = 7.6Hz, 1H), 8.50 (d, J = 2.8Hz, 1H), 7.70 (d, J = 2.0Hz, 1H), 7.54 - 7.49 (m, 1H), 7.49 - 7.45 (m, 1H), 5.71 (d, J = 9.2Hz, 1H), 5.20 - 5.14 (m, 1H), 4.47 - 4.41 (m, 1H), 4.24 - 4.18 (m, 2H), 4.09 - 3.94 (m, 2H), 3.80 - 3.61 (, 5H), 3.49 - 3.43 (m, 2H), 3.35 (s, 3H), 3.17 - 3.12 (m, 2H), 3.00 (s, 3H), 2.83 - 2.76 (m, 1H), 2.57 (d, J = 14.4Hz, 1H), 2.23 - 2.16 (m, 1H), 1.99 - 1.93 (m, 1H), 1.84 - 1.77 (m, 1H), 1.68 - 1.61 (m, 1H), 1.52 - 1.48 (m, 1H), 1.47 - 1.41 (m, 4H), 1.27 - 1.22 (m, 1H), 1.13 (d, J = 6.0Hz, 3H), 1.11 - 1.05 (m, 2H), 0.96 (s, 3H), 0.90 - 0.77 (m, 1H), 0.68 - 0.62 (m, 1H), 0.44 (s, 3H). Example 5 and Example 6 (1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[ (1S)-1-methoxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-1 5-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]cyclopropanecarboxamide and (1R,2R)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)- 20-[2-[(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyr idyl]-17,17-dimethyl-8,14- 2,5 9,13 22,26 dioxo-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxami de The compounds were prepared according to the following scheme:
Step 1: Preparation of (1S,2S)-2-cyano-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2- [(1S)-1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-15- 2,5 9,13 22,26 oxa-4-thia-9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaen-7-yl]cyclopropanecarboxami de and (1R,2R)-2-cyano-N- [(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethy l]-5-(4-methylpiperazin-1-yl)- 3-pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,2 8- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7- yl]cyclopropanecarboxamide To a solution of trans-2-Cyanocyclopropanecarboxylic acid (compound 5a, 11.2 mg, 0.1 mmol) in DMF (0.5 mL) were added DIEA (0.1 mL, 0.33 mmol), HATU (76.4 mg, 0.2 mmol) and (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thi a-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (intermediate D, 50.0 mg, 0.07 mmol) at 0 °C. After being stirred at 20 °C for 16 hrs, the reaction mixture was prepared by prep-HPLC and SFC to afford Example 5 (13.4 mg, faster eluted) as a white solid and Example 6 (9.9 mg, slower eluted) as a white solid. Example 5: MS calc’d 840.4 (MH + ), measured 840.1 (MH + ). 1 H NMR (400 MHz, CHLOROFORM-d) δ = 8.63 (d, J = 7.3 Hz, 1H), 8.49 (d, J = 2.8 Hz, 1H), 7.59 (d, J = 2.0 Hz, 1H), 7.12 - 7.07 (m, 2H), 6.88 (br d, J = 8.4 Hz, 1H), 5.92 (t, J = 8.9 Hz, 1H), 4.60 (br dd, J = 2.4, 12.5 Hz, 1H), 4.30 - 4.19 (m, 3H), 4.08 - 3.99 (m, 2H), 3.85 (br d, J = 11.1 Hz, 1H), 3.74 - 3.70 (m, 1H), 3.49 (br d, J = 3.4 Hz, 4H), 3.36 (s, 3H), 3.20 - 3.13 (m, 2H), 2.76 - 2.68 (m, 4H), 2.63 (s, 1H), 2.46 (br d, J = 14.2 Hz, 1H), 2.25 - 2.20 (m, 2H), 2.01 (s, 1H), 1.95 - 1.90 (m, 2H), 1.58 (td, J = 5.0, 9.8 Hz, 2H), 1.47 - 1.42 (m, 5H), 1.30 - 1.22 (m, 4H), 0.97 (t, J = 7.1 Hz, 3H), 0.91 (s, 2H), 0.47 (s, 3H) ppm. Example 6: MS calc’d 840.4 (MH + ), measured 840.1 (MH + ). 1 H NMR (400 MHz, CHLOROFORM-d) δ = 8.64 (d, J = 7.6 Hz, 1H), 8.51 (d, J = 2.9 Hz, 1H), 7.63 (d, J = 2.2 Hz, 1H), 7.13 - 7.07 (m, 2H), 6.88 (d, J = 9.5 Hz, 1H), 5.95 (t, J = 8.9 Hz, 1H), 4.65 - 4.57 (m, 1H), 4.32 - 4.21 (m, 3H), 4.12 - 3.96 (m, 2H), 3.86 (d, J = 10.8 Hz, 1H), 3.75 (d, J = 11.1 Hz, 1H), 3.48 (br d, J = 14.7 Hz, 1H), 3.37 (s, 3H), 3.34 - 3.29 (m, 4H), 3.19 (dd, J = 8.8, 15.0 Hz, 1H), 3.05 (br d, J = 14.2 Hz, 1H), 2.76 - 2.62 (m, 5H), 2.52 (br d, J = 14.3 Hz, 1H), 2.41 (s, 3H), 2.27 - 2.19 (m, 2H), 2.04 - 2.00 (m, 2H), 1.48 - 1.43 (m, 4H), 1.32 - 1.23 (m, 2H), 1.00 (t, J = 7.1 Hz, 3H), 0.92 (s, 4H), 0.49 (s, 3H) ppm. Example 7 (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-m ethoxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-15-oxa-4-thia- 2,5 9,13 22,26 9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23- hexaen-7-yl]-2-methyl-cyclopropanecarboxamide
The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-15- oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate F) instead of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D). Example 7 (12.5 mg) was obtained as a yellow solid. MS calc’d 897.4 (MH + ), measured 897.6 (MH + ). 1 H NMR (400 MHz, Methanol-d 4 ) δ = 8.68 (d, J = 7.6 Hz, 1H), 8.38 (d, J = 2.8 Hz, 1H), 7.78 (d, J = 2.4 Hz, 1H), 7.65 (d, J = 2.4 Hz, 1H), 7.35 (d, J = 12.8 Hz, 1H), 5.81 (d, J = 8.8 Hz, 1H), 4.48 - 4.40 (m, 1H), 4.38 - 4.31 (m, 1H), 4.30 - 4.19 (m, 1H), 4.16 - 4.01 (m, 2H), 3.81 - 3.68 (m, 2H), 3.49 - 3.44 (m, 4H), 3.43 - 3.40 (m, 1H), 3.37 (s, 3H), 3.21 - 3.12 (m, 3H), 3.04 - 3.95 (m, 1H), 2.92 - 2.86 (m, 4H), 2.81 - 2.70 (m, 2H), 2.21 - 2.11 (m, 1H), 1.99 - 1.89 (m, 1H), 1.84 - 1.70 (m, 1H), 1.67 - 1.57 (m, 1H), 1.51 - 1.46 (m, 1H), 1.44 (d, J = 6.4 Hz, 3H), 1.25 - 1.19 (m, 1H), 1.15 - 1.10 (m, 3H), 1.09 - 1.05 (m, 1H), 1.04 - 1.00 (m, 3H), 0.94 (s, 3H), 0.66 - 0.61 (m, 1H), 0.59 (s, 3H) ppm. Example 8 (1S,2S)-N-[(7S,13S)-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-m ethoxyethyl]-5-morpholino-3- pyridyl]-17,17-dimethyl-8,14-dioxo-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide
The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-morpholino-3- pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapent acyclo- 2,5 9,13 22,26 [17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) instead of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-(4- methylpiperazin-1-yl)-3-pyridyl]-17,17-dimethyl-15-oxa-4-thi a-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D). Example 8 (135.4 mg) was obtained as a light yellow solid. MS calc’d 816.4 (MH + ), measured 816.1 (MH + ). 1 H NMR (400 MHz, Methanol-d 4 ) δ = 8.70 (d, J = 7.5 Hz, 1H), 8.37 (d, J = 2.7 Hz, 1H), 7.95 (br s, 1H), 7.65 (d, J = 2.3 Hz, 1H), 7.37 (d, J = 12.6 Hz, 1H), 5.82 (br d, J = 8.9 Hz, 1H), 4.47 - 4.38 (m, 2H), 4.31 - 4.23 (m, 1H), 4.15 - 4.10 (m, 1H), 4.05 - 3.98 (m, 1H), 3.88 (br t, J = 4.7 Hz, 4H), 3.79 (br d, J = 11.0 Hz, 1H), 3.71 (d, J = 10.6 Hz, 1H), 3.47 - 3.38 (m, 10H), 3.00 (br d, J = 15.0 Hz, 1H), 2.84 - 2.72 (m, 2H), 2.19 - 2.12 (m, 1H), 1.97 - 1.90 (m, 1H), 1.64 (br d, J = 2.7 Hz, 1H), 1.50 - 1.45 (m, 4H), 1.23 (br dd, J = 4.2, 9.5 Hz, 2H), 1.12 (d, J = 5.9 Hz, 3H), 1.05 (q, J = 6.8 Hz, 4H), 0.96 (s, 3H), 0.62 (s, 3H) ppm. Example 9 (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyeth yl]-5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-8,1 4-dioxo-21-(2,2,2- 2,5 9,13 22,26 trifluoroethyl)-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[1 7.5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2-methyl-cyclopropan ecarboxamide
The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl] -5-[4-(2,2,2- trifluoroethyl)piperazin-1-yl]-3-pyridyl]-17,17-dimethyl-21- (2,2,2-trifluoroethyl)-15-oxa-4-thia- 2,5 9,13 22,26 9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene- 8,14-dione (Intermediate E) instead of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)- 1-methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17- dimethyl-15-oxa-4-thia- 2,5 9,13 22,26 9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene- 8,14-dione (Intermediate D). Example 9 mg) was obtained as an off-white solid. MS calc’d 951.4 (MH + ), measured 951.2 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.69 (d, J = 7.6 Hz, 1H), 8.42 (d, J = 2.8 Hz, 1H), 7.69 (d, J = 2.4 Hz, 1H), 7.56 - 7.43 (m, 2H), 5.73 (d, J = 8.8 Hz, 1H), 5.24 - 5.08 (m, 1H), 4.83 - 4.78 (m, 1H), 4.49 - 4.39 (m, 1H), 4.26 - 4.15 (m, 2H), 3.82 - 3.65 (m, 2H), 3.48 - 3.38 (m, 5H), 3.36 (s, 3H), 3.30 - 3.25 (m, 1H), 3.20 - 3.09 (m, 3H), 2.88 (t, J = 4.8 Hz, 4H), 2.84 - 2.75 (m, 1H), 2.67 - 2.59 (m, 1H), 2.26 - 2.15 (m, 1H), 1.99 - 1.91 (m, 1H), 1.87 - 1.74 (m, 1H), 1.70 - 1.56 (m, 1H), 1.52 - 1.47 (m, 1H), 1.45 (d, J = 6.0 Hz, 3H), 1.28 - 1.19 (m, 1H), 1.15 - 1.10 (m, 3H), 1.10 - 1.05 (m, 1H), 0.97 (s, 3H), 0.71 - 0.59 (m, 1H), 0.49 (s, 3H) ppm. Example 10 (1S,2S)-N-[(7S,13S)-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyeth yl]-5-(4-methylpiperazin-1- yl)-3-pyridyl]-17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoro ethyl)-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide
The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-25-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl] -5-(4-methylpiperazin-1-yl)-3- pyridyl]-17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-t hia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (intermediate H) instead of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D). Example 10 (11.2 mg) was obtained as a white solid. MS calc’d 883.4 (MH + ), measured 883.5 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.50 (d, J = 2.8 Hz, 1H), 7.52 - 7.37 (m, 4H), 6.03 – 5.88 (m, 1H), 5.19 - 5.07 (m, 1H), 4.80 - 4.76 (m, 2H), 4.44 - 4.34 (m, 2H), 4.12 - 3.96 (m, 3H), 3.72 - 3.47 (m, 6H), 3.41 - 3.33 (m, 2H), 3.28 - 3.25 (m, 2H), 3.19 - 3.05 (m, 5H), 2.99 (s, 4H), 2.59 - 2.52 (m, 1H), 1.66 - 1.53 (m, 1H), 1.44 (d, J = 6.0 Hz, 3H), 1.39 - 1.29 (m, 2H), 1.26 - 1.17 (m, 2H), 1.10 - 1.05 (m, 3H), 1.05 - 1.01 (m, 1H), 0.80 (s, 3H), 0.65 - 0.52 (m, 3H) ppm. Example 11 (1S,2S)-2-(difluoromethyl)-N-[(7S,13S)-21-ethyl-24-fluoro-(2 0M)-20-[2-[(1S)-1- methoxyethyl]-5-morpholino-3-pyridyl]-17,17-dimethyl-8,14-di oxo-15-oxa-4-thia- 2,5 9,13 22,26 9,21,27,28-tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23- hexaen-7-yl]cyclopropanecarboxamide
The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-meth oxyethyl]-5-morpholino-3- pyridyl]-17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapent acyclo- 2,5 9,13 22,26 [17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate G) and (1S,2S)-2-(difluoromethyl)cyclopropanecarboxylic acid instead of (7S,13S)-7-amino-21- ethyl-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl]-5-(4-methy lpiperazin-1-yl)-3-pyridyl]- 2,5 9,13 22,26 17,17-dimethyl-15-oxa-4-thia-9,21,27,28-tetrazapentacyclo[17 .5.2.1 .1 .0 ]octacosa- 1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D) and (1S,2S)-2- methylcyclopropanecarboxylic acid (compound 1a). Example 11 (33.2 mg) was obtained as a yellow solid. MS calc’d 852.4 (MH + ), measured 852.2 (MH + ). 1 H NMR (400 MHz, CHLOROFORM-d) δ = 8.91 (br s, 1H), 8.66 (d, J = 7.6 Hz, 1H), 7.62 (d, J = 2.2 Hz, 1H), 7.41 (s, 1H), 7.13 (s, 1H), 7.10 (s, 1H), 6.75 (br d, J = 8.9 Hz, 1H), 5.99 - 5.93 (m, 1H), 5.83 (d, J = 3.1 Hz, 1H), 4.60 (br d, J = 11.9 Hz, 1H), 4.38 (br d, J = 6.4 Hz, 1H), 4.21 (br d, J = 9.2 Hz, 1H), 4.13 - 4.06 (m, 2H), 3.87 (br d, J = 11.4 Hz, 2H), 3.79 - 3.65 (m, 2H), 3.45 (br d, J = 15.2 Hz, 1H), 3.41 (s, 2H), 3.39 - 3.35 (m, 4H), 3.17 (br dd, J = 8.6, 15.0 Hz, 2H), 3.10 (br d, J = 11.6 Hz, 1H), 2.80 - 2.63 (m, 2H), 2.50 (br d, J = 13.9 Hz, 1H), 2.20 (br d, J = 10.9 Hz, 1H), 1.99 (br d, J = 17.9 Hz, 1H), 1.87 - 1.78 (m, 2H), 1.67 - 1.59 (m, 1H), 1.53 (d, J = 6.4 Hz, 3H), 1.33 - 1.21 (m, 3H), 1.18 - 1.12 (m, 1H), 1.01 (br t, J = 7.0 Hz, 3H), 0.94 (s, 3H), 0.53 (s, 3H) ppm. Example 12 (1S,2S)-N-[(7S,13S)-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyeth yl]-5-morpholino-3-pyridyl]- 17,17-dimethyl-8,14-dioxo-21-(2,2,2-trifluoroethyl)-15-oxa-4 -thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaen-7-yl]-2- methyl-cyclopropanecarboxamide The title compound was prepared in analogy to the preparation of Example 1 by using (7S,13S)-7-amino-24-fluoro-(20M)-20-[2-[(1S)-1-methoxyethyl] -5-morpholino-3-pyridyl]- 17,17-dimethyl-21-(2,2,2-trifluoroethyl)-15-oxa-4-thia-9,21, 27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (intermediate I) instead of (7S,13S)-7-amino-21-ethyl-24-fluoro-(20M)-20-[2-[(1S)-1- methoxyethyl]-5-(4-methylpiperazin-1-yl)-3-pyridyl]-17,17-di methyl-15-oxa-4-thia-9,21,27,28- 2,5 9,13 22,26 tetrazapentacyclo[17.5.2.1 .1 .0 ]octacosa-1(25),2,5(28),19,22(26),23-hexaene-8,14-dione (Intermediate D). Example 12 (150 mg) was obtained as a white solid. MS calc’d 870.4 (MH + ), measured 870.5 (MH + ). 1 H NMR (400 MHz, Methanol-d4) δ = 8.65-8.63 (d, J = 7.6 Hz, 1H), 8.41 - 8.40 (d, J = 2.8 Hz,1H), 7.70 - 7.65 (d, J = 2.4 Hz, 1H), 7.50-7.40 (d, J = 12.4 Hz, 1H), 7.31 - 7.28 (d, J = 2.4 Hz, 1H), 5.76 - 5.66 (d, J = 8.0Hz,1H), 5.18 - 5.05 (m, 1H), 4.96 - 4.88 (m, 1H), 4.47 - 4.39 (m, 1H), 4.23 - 4.10 (m, 2H), 3.90-3.83 (t, J = 4.8 Hz, J = 9.6 Hz, 4H), 3.80- 3.75 (d, J = 10.8Hz, 1H), 3.71 - 3.63 (m, 1H), 3.49 - 3.42 (m, 1H), 3.30 - 3.29 (m, 3H), 3.28 - 3.21 (m, 4H), 3.14 - 3.07 (d, J = 14.4 Hz, 1H), 2.86 - 2.77 (m, 1H), 2.63-2.55 (d, J = 14.4 Hz, 1H), 2.23 - 2.14 (m, 1H), 1.98 - 1.89 (m, 1H), 1.84 - 1.72 (m, 1H), 1.65 - 1.56 (m, 1H), 1.52 - 1.46 (m, 1H),1.45- 1.40 (d, J = 6.4 Hz, 3H), 1.30 - 1.19 (m, 2H), 1.13 - 1.06 (m, 4H), 0.95 (s, 3H), 0.69 - 0.58 (m, 1H), 0.48 - 0.39 (s, 3H) ppm. BIOLOGICAL EXAMPLE Compound A122 (page 70 of Table.1a) from WO2022060836 was cited as reference compound for this invention. Example 13 Cell viability assay The purpose of this cellular assay was to determine the effects of test compounds on the proliferation of human cancer cell lines NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL- 1739) cells, SW620 (ATCC-CCL-227) over a 3-day treatment period by quantifying the amount of NADPH present at endpoint using Cell Counting Kit-8. Cells were seeded at 5,000 cells/well (NCI-H358), 2,000 cells/well (AGS) 2,000 cells/well (SW620) in 96-well assay plates (Corning-3699) and incubated overnight. On the day of the assay, diluted compounds were then added in a final concentration of 0.5% DMSO. After 72 hrs incubation, a tenth of the volume of cell counting kit 8(Dnjindo-CK04) was added into each well. Read the signal (OD450 minus OD650) using EnVision after 2 hrs incubation. IC 50 was determined by fitting a 4-parameter sigmoidal concentration response model. Table 1. Activity of Examples and Compounds of present invention in KRAS Cell viability assay
Example 14 KRAS-BRAF with CYPA (500 nM) interaction assay In this example, TR-FRET was also used to measure the compound or compound-CYPA dependent disruption of the KRAS G12C-BRAF complex. This protocol was also used to measure disruption of KRAS G12D or KRAS G12V binding to BRAF by a compound of the invention, respectively. In assay buffer containing 25mM HEPES PH=7.4 (4-(2-hydroxyethyl)- 1-piperazineethanesulfonic acid, Thermo, 15630080), 0.002% Tween20, 0.1% BSA, 100mM NaCl, 5mM MgCl2, 10 µM GMPPNP (Guanosine 5′-[β,γ-imido]triphosphate trisodium salt hydrate, Sigma, G0635), tagless CYPA, GMPPNP loaded 6His-KRAS proteins, and GST- BRAF RBD were mixed in a well of a 384-well assay plate at final concentrations of 50 nM, 6.25 nM and 1nM, respectively. Compound was present in plate wells as a 16-point 3-fold dilution series starting at a final concentration of 10 µM and incubated for 3 hours. A mixture of MAb Anti-6His-XL665 (Cisbio, 61HISXLB) and Mab anti-GST-TB cryptate (Cisbio, 61GSTTLB)was then added at a final concentration of 6.67 nM and 0.21 nM, respectively, and the plate was incubated for an additional 1.5 hours. TR-FRET signal was read on a PHERstar FSX microplate reader (Ex320 nm, Em 665/615 nm). Compounds that facilitate disruption of the KRAS-BRAF complex were identified as those eliciting a decrease in the TR-FRET ratio relative to DMSO control wells. Table 2. Activity of Examples and Compounds of present invention in KRAS-BRAF with CYPA (500 nM) interaction assay Example 15 pERK inhibition assay This assay is to measure the ability of test compounds in inhibiting the phosphorylation of ERK, the downstream signaling of KRAS G12C in NCI-H358 cells, KRAS G12D in AGS cells, and KRAS G12V in SW620. NCI-H358 (ATCC-CRL5807) cells, AGS (ATCC-CRL-1739) cells, SW620 (ATCC-CCL-227) cells were all grown and maintained using RPMI-1640 medium (Thermo Fisher Scientific) with 10% fetal bovine serum and 1% penicillin/streptomycin. On the day prior to compound addition, cells were plated in tissue culture-treated 96 well plates (Corning-3699) at a density of 30,000 cell/well, 20,000 cell/well, 30,000 cell/well for NCI-H358, AGS and SW620 respectively, and allowed for attachment overnight. Diluted compounds were then added in a final concentration of 0.5% DMSO. After 4 hours of incubation, the medium was removed, 100 µL of 4% formaldehyde was added, and the assay plates were incubated at room temperature for 20 minutes. The plates were then washed once with phosphate buffered saline (PBS), and permeabilized with 100 µL of chilled methanol for 10 minutes. Non-specific antibody binding to the plates was blocked using 50 µL 1X BSA blocking buffer (Thermo-37520, 10-fold dilution by Phosphate-Buffered Saline Tween (PBST) for at least 1 hour at room temperature. The amount of phosphor-ERK was determined using an antibody specific for phosphorylated form of ERK. Primary antibody (pERK, CST-4370, Cell Signaling Technology) was diluted 1:300 in blocking buffer, with 50 µL aliquoted to each well, and incubated overnight at 4 ℃. Cells was washed five times for 5 minutes with PBST. Secondary antibody (HRP-linked anti-rabbit IgG, CST-7074, Cell Signaling Technology) was diluted 1:1000 in blocking buffer, and 50 µL was added to each well and incubated 1-2 hrs at room temperature. Cells was washed 5 times for 5 minutes with PBST, 100µL TMB ELISA substrate (abcam-ab171523) were added and gently shake for 20 minutes.50µL stop solution (abcam-ab171529) were added, and then read the signal (OD450) by EnVision. IC50 was determined by fitting a 4-parameter sigmoidal concentration response model. Table 3. Activity of Examples and Compounds of present invention in KRAS pERK inhibition assay
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