Macrocyclic derivative and application thereof

Abstract

The invention discloses a macrocyclic derivative and application thereof, and particularly discloses a compound as shown in a formula (III), and a stereoisomer and pharmaceutically acceptable salt thereof. # imgabs0 #

Description

[0001] The present invention claims the following priority:

[0002] CN202211213222.2, application date: September 29, 2022;

[0003] CN2023101326996, application date: February 17, 2023;

[0004] CN2023103711659, application date: April 7, 2023;

[0005] CN2023106857489, application date: June 9, 2023;

[0006] CN2023109116322, application date: July 24, 2023.

[0007] This application is a divisional of the Chinese invention patent application with the application date of September 28, 2023, application number 202380053250.9, and invention name “Macrocyclic derivatives and their applications”. Technical Field

[0008] The present invention relates to a class of macrocyclic derivatives and applications thereof, and in particular to a compound represented by formula (VI), its stereoisomers and pharmaceutically acceptable salts thereof. Background Art

[0009] RAS (including KRAS, NRAS and HRAS) is downstream of growth factor receptors such as EGFR. It is a small GTPase of a type of molecular switch and a key node in the RAS-RAF-MEK-ERK signaling pathway and the PI3K-AKT-mTOR signaling pathway. It can regulate cell proliferation, survival and other events. RAS mutations can lead to functional activation of proteins by disrupting the GTP hydrolysis process. Under normal physiological conditions, RAS usually exists in the form of inactive RAS (OFF) bound to GDP; while in RAS mutated tumor cells, RAS mainly exists in the form of activated RAS (ON) bound to GTP. RAS mutated cancer patients account for 30% of all cancer patients (US data), of which KRAS, NRAS, and HRAS mutations account for 85%, 11%, and 4% respectively. Each RAS has nearly 20 mutations. For example, KRAS has KRAS G12C 、KRAS G12D 、KRAS G12V 、KRAS G12R 、KRAS G13C Since RAS mutation is an important factor in the occurrence and development of cancer, mutant RAS has become an important target for cancer treatment.G12C The covalent inhibitors Sotorasib and Adagrasib in KRAS G12C Great success in non-small cell lung cancer, but KRAS G12D 、KRAS G12V 、KRAS G13 There is still no effective targeted therapy for cancers with other KRAS mutation types and other RAS mutation types such as NRAS. Therefore, the development of broad-spectrum RAS inhibitors is of great clinical significance.

[0010] Revolution Medicines, Inc. has announced a class of macrocyclic compounds (WO2020132597, WO2021091982, WO2021091967, WO2021091956). This class of compounds can form a ternary complex with the chaperone proteins CypA and RAS (ON) in the body. The formation of the ternary complex blocks the binding of RAF downstream of RAS, thereby inhibiting the RAS-RAF-MEK-ERK signaling pathway and achieving an anti-tumor effect. This type of RAS inhibitor has a good inhibitory effect on different RAS mutations and RAS-dependent tumors. The design and synthesis of broad-spectrum RAS inhibitors with excellent drug properties based on this type of macrocyclic compounds has great clinical application value. Summary of the invention

[0011] The present invention provides a compound represented by formula (VI), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0012]

[0013] L is R6 or

[0014] L1 is selected from -N(R9)C(=O)-;

[0015] L2 is selected from C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R a replace;

[0016] L3 is selected from -CH2- and C 3-6 Cycloalkyl;

[0017] L4 is selected from a single bond and -C 1-4 Alkyl-N(R 10 )C(=O)-;

[0018] Ring A is selected from tetrahydropyridazinyl, 3,4-diazabicyclo[4.1.0]heptyl, 2,3-diazabicyclo[3.1.0]hexanyl, 5,6-diazaspiro[2.5]octanyl, 3,4-diazabicyclo[4.2.0]octanyl and 2,3-diazabicyclo[3.1.1]heptyl;

[0019] Ring B is selected from 5-membered heteroaryl, 5-membered heteroaryl, indolyl and

[0020] T1, T2, T3 and T2 are each independently selected from CH and N;

[0021] R1 is selected from H, F, Cl, Br, I, OH, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R h replace;

[0022] R2 is selected from -O- and -NH-;

[0023] R3 is selected from phenyl and 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b replace;

[0024] R4 and R5 are independently selected from H, C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R c replace;

[0025] R6 is selected from C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R d replace;

[0026] Each R7 is independently selected from H, halogen, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R e replace;

[0027] Each R8 is independently selected from H, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6Cycloalkyl and 3-10 membered heterocycloalkyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-10 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R f replace;

[0028] R9, R 10 Selected from H and C 1-4 Alkyl, the C 1-4 The alkyl group is optionally substituted with 1, 2 or 3 R g replace;

[0029] Each R a , each R b , each R c , each R e , each R f , each R g and each R h are independently selected from H, D, F, Cl, Br, I, OH, C 1-3 Alkyl and C 1-3 Alkoxy, the C 1-3 Alkyl and C 1-3 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R;

[0030] Each R d are independently selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 Alkenyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 The alkenyl group is optionally substituted with 1, 2 or 3 R groups, each independently;

[0031] Each R is independently selected from D, F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3;

[0032] n, p and q are independently selected from 0, 1, 2 and 3;

[0033] The “3-6 membered heterocycloalkyl”, “3-10 membered heterocycloalkyl” and “5-6 membered heteroaryl” each independently contain 1 or 2 heteroatoms or heteroatom groups independently selected from —NH—, —O—, —S— and N.

[0034] The present invention also provides a compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0035]

[0036] L is R6 or

[0037] L1 is selected from -N(R9)C(=O)-;

[0038] L2 is selected from C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R a replace;

[0039] L3 is selected from -CH2- and C 3-6 Cycloalkyl;

[0040] L4 is selected from a single bond and -C 1-4 Alkyl-N(R 10 )C(=O)-;

[0041] Ring A is selected from tetrahydropyridazinyl, 3,4-diazabicyclo[4.1.0]heptyl, 2,3-diazabicyclo[3.1.0]hexanyl, 5,6-diazaspiro[2.5]octanyl, 3,4-diazabicyclo[4.2.0]octanyl and 2,3-diazabicyclo[3.1.1]heptyl;

[0042] Ring B is selected from 5-membered heteroaryl, 5-membered heteroaryl, indolyl and

[0043] R1 is selected from H, F, Cl, Br, I, OH, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R h replace;

[0044] R2 is selected from -O- and -NH-;

[0045] R3 is selected from phenyl and 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b replace;

[0046] R4 and R5 are independently selected from H, C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R c replace;

[0047] R6 is selected from C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R d replace;

[0048] Each R7 is independently selected from H, halogen, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R e replace;

[0049] Each R8 is independently selected from H, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-7 membered heterocycloalkyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R f replace;

[0050] R9, R 10 Selected from H and C 1-4 Alkyl, the C 1-4 The alkyl group is optionally substituted with 1, 2 or 3 R g replace;

[0051] Each R a , each R b , each R c , each R e , each R f , each R g and each R h Each independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3;

[0052] Each R d are independently selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 Alkenyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 The alkenyl group is optionally substituted with 1, 2 or 3 R groups, each independently;

[0053] Each R is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3;

[0054] n, p and q are independently selected from 0, 1, 2 and 3;

[0055] The “3-6 membered heterocycloalkyl”, “3-7 membered heterocycloalkyl” and “5-6 membered heteroaryl” each independently contain 1 or 2 heteroatoms or heteroatom groups independently selected from —NH—, —O—, —S— and N.

[0056] The present invention also provides a compound represented by formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0057]

[0058] L1 is selected from -N(R9)C(=O)-;

[0059] L2 is selected from C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R a replace;

[0060] L3 is selected from -CH2-;

[0061] L4 is selected from a single bond and -C 1-4 Alkyl-N(R 10 )C(=O)-;

[0062] Ring A is selected from tetrahydropyridazinyl, 3,4-diazabicyclo[4.1.0]heptane, 2,3-diazabicyclo[3.1.0]hexane and 5,6-diazaspiro[2.5]octane;

[0063] R1 is selected from H, F, Cl, Br, I, OH, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R h replace;

[0064] R2 is selected from -O- and -NH-;

[0065] R3 is selected from phenyl and 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b replace;

[0066] R4 and R5 are independently selected from H, C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R c replace;

[0067] R6 is selected from C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R d replace;

[0068] R7 is selected from H, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R e replace;

[0069] R8 is selected from H, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-7 membered heterocycloalkyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R f replace;

[0070] R9, R 10 Selected from H and C 1-4 Alkyl, the C 1-4 The alkyl group is optionally substituted with 1, 2 or 3 R g Replacement; each R a , R b , R c , R e , R f , R g and R h Each independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3;

[0071] Each R d are independently selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 Alkenyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 The alkenyl group is optionally substituted with 1, 2 or 3 R groups, each independently;

[0072] Each R is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3;

[0073] n, p and q are independently selected from 0, 1, 2 and 3;

[0074] The “3-6 membered heterocycloalkyl”, “3-7 membered heterocycloalkyl” and “5-6 membered heteroaryl” each independently contain 1 or 2 heteroatoms or heteroatom groups independently selected from —NH—, —O—, —S— and N.

[0075] In some embodiments of the present invention, each R mentioned above is independently selected from D and F, and other variables are as defined in the present invention.

[0076] In some embodiments of the present invention, each R mentioned above is independently selected from F, and other variables are as defined in the present invention.

[0077] In some embodiments of the present invention, each of the above R a are independently selected from H, D, F, Cl and CH3, and other variables are as defined in the present invention.

[0078] In some embodiments of the present invention, each of the above R b are independently selected from H, D, F, Cl, OH and CH3, and other variables are as defined in the present invention.

[0079] In some embodiments of the present invention, each of the above R c are independently selected from H, D, F and Cl, and other variables are as defined in the present invention.

[0080] In some embodiments of the present invention, each of the above R d are independently selected from CH3, CH2CH3, phenyl, cyclopropyl, cyclobutyl, oxetanyl, azetidinyl and The CH3, CH2CH3, phenyl, cyclopropyl, cyclobutyl, oxetanyl, azetidinyl and Each is independently optionally substituted with 1, 2 or 3 R, and the other variables are as defined herein.

[0081] In some embodiments of the present invention, each of the above R d are independently selected from CH3, Other variables are as defined herein.

[0082] In some embodiments of the present invention, each of the above R d are independently selected from C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R groups, and the other variables are as defined herein.

[0083] In some embodiments of the present invention, each of the above R dare independently selected from CH3 and CH2CH3, said CH3 and CH2CH3 are independently optionally substituted with 1, 2 or 3 F, and other variables are as defined herein.

[0084] In some embodiments of the present invention, each of the above R d are each independently selected from CH3, wherein CH3 is optionally substituted with 1, 2 or 3 F, and other variables are as defined herein.

[0085] In some embodiments of the present invention, each of the above R d are independently selected from CH3, and other variables are as defined in the present invention.

[0086] In some embodiments of the present invention, each of the above R d are independently selected from CH3, and other variables are as defined in the present invention.

[0087] In some embodiments of the present invention, each of the above R e are independently selected from H, D, F and Cl, and other variables are as defined in the present invention.

[0088] In some embodiments of the present invention, each of the above R e are independently selected from D and F, and other variables are as defined in the present invention.

[0089] In some embodiments of the present invention, each of the above R f are independently selected from H, D, F, Cl, CH3 and OCH3, said CH3 and OCH3 are independently optionally substituted by 1, 2 or 3 R, and other variables are as defined in the present invention.

[0090] In some embodiments of the present invention, each of the above R f are independently selected from H, D, F, Cl, CH3, CD3, CF3 and OCH3, and other variables are as defined in the present invention.

[0091] In some embodiments of the present invention, each of the above R g are independently selected from H, D, F and Cl, and other variables are as defined in the present invention.

[0092] In some embodiments of the present invention, each of the above R h are independently selected from H, D, F and Cl, and other variables are as defined in the present invention.

[0093] In some embodiments of the present invention, each R1 mentioned above is independently selected from H, F, OH and CH3, and other variables are as defined in the present invention.

[0094] In some embodiments of the present invention, each R1 mentioned above is independently selected from H, F and CH3, and other variables are as defined in the present invention.

[0095] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0096] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0097] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0098] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0099] In some embodiments of the present invention, the ring A is selected from 3,4-diazabicyclo[4.1.0]heptyl and 2,3-diazabicyclo[3.1.1]heptyl, and other variables are as defined in the present invention.

[0100] In some embodiments of the present invention, the ring A is selected from Other variables are as defined herein.

[0101] In some embodiments of the present invention, the ring A is Other variables are as defined herein.

[0102] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0103] In some embodiments of the present invention, the above R2 is selected from -O-, and other variables are as defined in the present invention.

[0104] In some embodiments of the present invention, the above R3 is selected from 5-6 membered heteroaryl, wherein the 5-6 membered heteroaryl is optionally substituted by 1, 2 or 3 R b The other variables are as defined in the present invention.

[0105] In some embodiments of the present invention, the above R3 is selected from a 5-membered heteroaryl group, wherein the 5-membered heteroaryl group is optionally replaced by 1, 2 or 3 R b The other variables are as defined in the present invention.

[0106] In some embodiments of the present invention, the above R3 is selected from thiazolyl, thienyl, oxazolyl, pyrazolyl and imidazolyl, and the thiazolyl, thienyl, oxazolyl, pyrazolyl and imidazolyl are independently optionally substituted by 1, 2 or 3 R b The other variables are as defined in the present invention.

[0107] In some embodiments of the present invention, the above R3 is selected from thiazolyl and thienyl, and other variables are as defined in the present invention.

[0108] In some embodiments of the present invention, the above R3 is selected from Other variables are as defined herein.

[0109] In some embodiments of the present invention, the above R3 is selected from Other variables are as defined herein.

[0110] In some embodiments of the present invention, the above R3 is selected from phenyl, pyridyl, pyrimidyl, thiazolyl, thienyl, oxazolyl, pyrazolyl and imidazolyl, and the phenyl, pyridyl, pyrimidyl, thiazolyl, thienyl, oxazolyl, pyrazolyl and imidazolyl are independently optionally substituted by 1, 2 or 3 R b The other variables are as defined in the present invention.

[0111] In some embodiments of the present invention, the above R3 is selected from phenyl, pyridyl, pyrimidyl, thiazolyl, oxazolyl, pyrazolyl and imidazolyl, and the phenyl, pyridyl, pyrimidyl, thiazolyl, oxazolyl, pyrazolyl and imidazolyl are independently optionally substituted by 1, 2 or 3 R b The other variables are as defined in the present invention.

[0112] In some embodiments of the present invention, the above R3 is selected from Other variables are as defined herein.

[0113] In some embodiments of the present invention, the above R3 is selected from Other variables are as defined herein.

[0114] In some embodiments of the present invention, the above R3 is selected from Other variables are as defined herein.

[0115] In some embodiments of the present invention, the above R4 is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl and cyclopentyl, and the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl and cyclopentyl are independently optionally replaced by 1, 2 or 3 R c The other variables are as defined in the present invention.

[0116] In some embodiments of the present invention, the above R5 is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl and cyclopentyl, and the CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl and cyclopentyl are independently optionally replaced by 1, 2 or 3 R c The other variables are as defined in the present invention.

[0117] In some embodiments of the present invention, R4 is selected from H, R5 is selected from H, CH(CH3)2 and Other variables are as defined herein.

[0118] In some embodiments of the present invention, the above R6 is selected from cyclopentyl, aziridine, azetidin, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl and oxhexyl, and the cyclopentyl, aziridine, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl and oxhexyl are independently and optionally replaced by 1, 2 or 3 R d The other variables are as defined in the present invention.

[0119] In some embodiments of the present invention, the above R6 is selected from cyclopropyl, cyclobutyl, cyclopentyl, aziridine, azetidin, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl and oxhexyl, and the cyclopropyl, cyclobutyl, cyclopentyl, aziridine, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl and oxhexyl are independently and optionally replaced by 1, 2 or 3 R d The other variables are as defined in the present invention.

[0120] In some embodiments of the present invention, the above R6 is selected from Other variables are as defined herein.

[0121] In some embodiments of the present invention, the above R6 is selected from Other variables are as defined herein.

[0122] In some embodiments of the present invention, the above R6 is selected from C 3-6 Cycloalkyl, the C 3-6 The cycloalkyl group is optionally substituted with 1, 2 or 3 R d The other variables are as defined in the present invention.

[0123] In some embodiments of the present invention, the above R6 is selected from cyclopropyl, and the cyclopropyl is optionally substituted by 1, 2 or 3 R d The other variables are as defined in the present invention.

[0124] In some embodiments of the present invention, the above R6 is selected from Other variables are as defined herein.

[0125] In some embodiments of the present invention, the above R6 is selected from Other variables are as defined herein.

[0126] In some embodiments of the present invention, each R7 is independently selected from H, F, Cl and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R e The other variables are as defined in the present invention.

[0127] In some embodiments of the present invention, the above R7 is selected from H, F, Cl, CH3 and CH2CH3, and the CH3 and CH2CH3 are independently optionally replaced by 1, 2 or 3 R e The other variables are as defined in the present invention.

[0128] In some embodiments of the present invention, the above R7 is selected from H, F, Cl, CH3, CH2CH3, CH2F, CHF2, CF3, CH2CF3 and CD3, and other variables are as defined in the present invention.

[0129] In some embodiments of the present invention, the above R7 is selected from H, F, Cl, CH3 and CH2CH3, and other variables are as defined in the present invention.

[0130] In some embodiments of the present invention, the above R7 is selected from H, CH3 and CH2CH3, and other variables are as defined in the present invention.

[0131] In some embodiments of the present invention, each R8 is independently selected from H, C 1-3 Alkyl and 3-10 membered heterocycloalkyl, the C 1-3 The alkyl group and the 3-10 membered heterocycloalkyl group are each independently optionally substituted with 1, 2 or 3 R f The other variables are as defined in the present invention.

[0132] In some embodiments of the present invention, each R8 is independently selected from H, C 1-3 Alkyl and 5-10 membered heterocycloalkyl, the C 1-3 The alkyl group and the 5-10 membered heterocycloalkyl group are each independently optionally substituted by 1, 2 or 3 R f The other variables are as defined in the present invention.

[0133] In some embodiments of the present invention, each R8 is independently selected from H, C 1-3 Alkyl and 5-6 membered heterocycloalkyl, the C 1-3 The alkyl and 5-6 membered heterocycloalkyl groups are each independently optionally substituted with 1, 2 or 3 Rf The other variables are as defined in the present invention.

[0134] In some embodiments of the present invention, the above R8 is selected from H, CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, tetrahydropyridinyl, homopiperidinyl, morpholinyl, The CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, tetrahydropyridinyl, homopiperidinyl, morpholinyl, are independently optionally substituted by 1, 2 or 3 R f The other variables are as defined in the present invention.

[0135] In some embodiments of the present invention, the above R8 is selected from H, Other variables are as defined herein.

[0136] In some embodiments of the present invention, the above R8 is selected from H, CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, homopiperidinyl, morpholinyl and The CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, homopiperidinyl, morpholinyl and are independently optionally substituted by 1, 2 or 3 R f The other variables are as defined in the present invention.

[0137] In some embodiments of the present invention, the above R8 is selected from H, CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, homopiperidinyl and morpholinyl, and the CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, homopiperidinyl and morpholinyl are independently and optionally substituted by 1, 2 or 3 R f The other variables are as defined in the present invention.

[0138] In some embodiments of the present invention, the above R8 is selected from H, Other variables are as defined herein.

[0139] In some embodiments of the present invention, the above R8 is selected from H, Other variables are as defined herein.

[0140] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0141] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0142] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0143] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0144] In some embodiments of the present invention, the above structural unit Selected from

[0145] Other variables are as defined herein.

[0146] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0147] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0148] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0149] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0150] In some embodiments of the present invention, the above structural unit Selected from Other variables are as defined herein.

[0151] In some embodiments of the present invention, the ring B is selected from Other variables are as defined herein.

[0152] In some embodiments of the present invention, the above-mentioned ring B is selected from indolyl, and other variables are as defined in the present invention.

[0153] In some embodiments of the present invention, the ring B is selected from Other variables are as defined herein.

[0154] In some embodiments of the present invention, the above L1 is selected from -N(CH3)C(=O)-, and other variables are as defined in the present invention.

[0155] In some embodiments of the present invention, the above L2 is selected from Other variables are as defined herein.

[0156] In some embodiments of the present invention, the above L3 is selected from -CH2-, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, and other variables are as defined in the present invention.

[0157] In some embodiments of the present invention, the above L3 is selected from -CH2-, Other variables are as defined herein.

[0158] In some embodiments of the present invention, the above L3 is selected from -CH2-, and other variables are as defined in the present invention.

[0159] In some embodiments of the present invention, the above L3 is selected from Other variables are as defined in the present invention. In some embodiments of the present invention, L4 is selected from a single bond and -CH 2- N(CH3)C(=O)-, and other variables are as defined in the present invention.

[0160] In some embodiments of the present invention, the above L is selected from Other variables are as defined herein.

[0161] In some embodiments of the present invention, the above L is selected from Other variables are as defined herein.

[0162] In some embodiments of the present invention, the above L is R6, and other variables are as defined in the present invention.

[0163] In some embodiments of the present invention, the above L is selected from Other variables are as defined herein.

[0164] In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt is selected from:

[0165]

[0166] wherein ring A, T1, T3, R1, R3, R6, R7, R8, L3 and n are as defined in the present invention.

[0167] In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt is selected from:

[0168]

[0169] in,

[0170] Structural unit Selected from

[0171] Structural unit Selected from

[0172] R6 is selected from C 3-6 Cycloalkyl, the C 3-6 The cycloalkyl group is optionally substituted with 1, 2 or 3 R d replace;

[0173] Each R d are independently selected from C 1-4 Alkyl, the C 1-4 The alkyl group is optionally substituted with 1, 2 or 3 R;

[0174] L3, R3, R7, each R8 and each R are as defined in the present invention.

[0175] In some embodiments of the present invention, the compound of formula (VI-1), formula (VI-2) or formula (VI-3), its stereoisomers or pharmaceutically acceptable salts thereof, wherein the structural unit Selected from Other variables are as defined herein.

[0176] In some embodiments of the present invention, the compound of formula (VI-1), its stereoisomer or a pharmaceutically acceptable salt thereof is selected from:

[0177]

[0178] in,

[0179] T1 and T3 are independently selected from CH and N;

[0180] Each R1 is independently selected from H, F, OH and CH3;

[0181] R3 is selected from phenyl and 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b replace;

[0182] R6 is selected from C 3-6 Cycloalkyl, the C 3-6 The cycloalkyl group is optionally substituted with 1, 2 or 3 R d replace;

[0183] R7 is selected from H and C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R e replace;

[0184] Each R8 is independently selected from H, C 1-3 Alkyl and 5-10 membered heterocycloalkyl, the C 1-3 The alkyl group and the 5-10 membered heterocycloalkyl group are each independently optionally substituted by 1, 2 or 3 R f replace;

[0185] L3 is selected from -CH2-, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl;

[0186] Each R d are independently selected from C 1-3 Alkyl, the C 1-3 The alkyl group is optionally substituted with 1, 2 or 3 R;

[0187] Each R b Each independently selected from H, D, F, Cl, OH and CH3;

[0188] Each R e are independently selected from H, D, F and Cl;

[0189] Each R f Each is independently selected from H, D, F, Cl, CH3 and OCH3, wherein CH3 and OCH3 are independently optionally substituted by 1, 2 or 3 R;

[0190] Each R is independently selected from D and F.

[0191] In some embodiments of the present invention, the above-mentioned compound of formula (P-1) or (P-2), its stereoisomer or a pharmaceutically acceptable salt thereof, wherein each R1, R3, R6, R7, each R8, T1, T3 and L3 are as defined in formula (VI), formula (III) or formula (I) of the present invention.

[0192] In some embodiments of the present invention, the compound of the above formula (P-1) or (P-2), its stereoisomers or pharmaceutically acceptable salts thereof, wherein R3 is a 5-6 membered heteroaryl group, and the other variables are as defined in the present invention.

[0193] In some embodiments of the present invention, the compound of the above formula (P-1) or (P-2), its stereoisomers or pharmaceutically acceptable salts thereof, wherein R3 is thienyl and thiazolyl, and other variables are as defined in the present invention.

[0194] In some embodiments of the present invention, the compound of formula (P-1) or (P-2), its stereoisomer or its pharmaceutically acceptable salt, wherein each R8 is independently selected from H, C 1-3 Alkyl and 5-6 membered heterocycloalkyl, the C 1-3 The alkyl and 5-6 membered heterocycloalkyl groups are each independently optionally substituted with 1, 2 or 3 R fThe other variables are as defined in the present invention.

[0195] In some embodiments of the present invention, the compound of formula (P-1) or (P-2), its stereoisomers or pharmaceutically acceptable salts thereof, wherein T1 is CH, T3 is CH, and other variables are as defined in the present invention.

[0196] In some embodiments of the present invention, the compound of formula (P-1) or (P-2), its stereoisomer or its pharmaceutically acceptable salt, wherein L3 is selected from -CH2-, Other variables are as defined herein.

[0197] In some embodiments of the present invention, the compound of formula (P-1) or (P-2), its stereoisomer or a pharmaceutically acceptable salt thereof is selected from:

[0198]

[0199]

[0200] Wherein, each R1, R3, R6, R7, each R8, T1, T3 and L3 are as defined in formula (P-1) or (P-2) of the present invention. In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt is selected from,

[0201]

[0202] wherein R1, R3, R4, R5, R6, R7, R8, L1, L3, L4 and n are as defined in the present invention.

[0203] In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt is selected from:

[0204]

[0205] wherein R1, R3, R6, R7 and R8 are as defined in the present invention.

[0206] In some embodiments of the present invention, the compound of formula (IV), its stereoisomer or a pharmaceutically acceptable salt thereof is selected from:

[0207]

[0208] wherein R1, R3, R6, R7 and R8 are as defined in the present invention.

[0209] In some embodiments of the present invention, the compound of formula (IV-1a), formula (IV-1b), its stereoisomer or pharmaceutically acceptable salt thereof is selected from:

[0210]

[0211] wherein R1, R3, R6, R7 and R8 are as defined in the present invention.

[0212] In some embodiments of the present invention, the compounds of the above formula (IV), (IV-1a), (IV-1b), (IV-1a-1), (IV-1a-2), (IV-1b-1), (IV-1b-2), their stereoisomers or pharmaceutically acceptable salts thereof, wherein R1 is selected from H, F, OH and CH3; R3 is selected from phenyl and thiazolyl, and the phenyl and thiazolyl are independently optionally substituted by 1, 2 or 3 R b Substituted; R6 is selected from cyclopropyl, the cyclopropyl is optionally substituted by 1, 2 or 3 R d R7 is selected from H, F, Cl, CH3 and CH2CH3; R8 is selected from H, Each R b Each R is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3; d are independently selected from CH3,

[0213] Some other solutions of the present invention are obtained by any combination of the above variables.

[0214] The present invention also provides the following compounds, their stereoisomers or pharmaceutically acceptable salts thereof:

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221] In some embodiments of the present invention, the above-mentioned compound, its stereoisomer or its pharmaceutically acceptable salt is selected from:

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240] The present invention also provides the use of the above compound, its stereoisomer or its pharmaceutically acceptable salt in the preparation of RAS inhibitor drugs.

[0241] The RAS inhibitor drug of the present invention is used to treat RAS mutations and RAS-dependent tumors, such as solid tumors; further, the solid tumor is pancreatic cancer, lung cancer or colorectal cancer.

[0242] The present invention also provides the following synthesis method:

[0243] Method 1 - Intermediate:

[0244]

[0245] Method 2 - Intermediate:

[0246]

[0247] Method 3 - Intermediate:

[0248]

[0249] Method 4 - Intermediate:

[0250]

[0251] Method 5 - Intermediate:

[0252]

[0253] Method 6 - Intermediate:

[0254]

[0255] Method 7 - Intermediate

[0256]

[0257] Method 8 - Intermediate

[0258]

[0259] Method 9 - Intermediate

[0260]

[0261] Method 10 - Intermediate

[0262]

[0263] Method 11:

[0264]

[0265] Method 12:

[0266]

[0267] Method 13:

[0268]

[0269]

[0270] Method 14:

[0271] Method 15:

[0272]

[0273]

[0274] Method 16:

[0275]

[0276]

[0277] Method 17:

[0278]

[0279] Method 18:

[0280]

[0281]

[0282] Technical Effects

[0283] The compounds of the present invention have a good binding effect with the chaperone protein CypA. The binding with CypA will block the binding of RAF downstream of RAS with RAS, thereby inhibiting the RAS-RAF-MEK-ERK signaling pathway to achieve an anti-tumor effect; the compounds of the present invention have significant inhibitory activity on the proliferation of RAS mutant cell lines (such as GP2D, PK-59, AsPC-1, PSN-1, RKN, Capan-1, SW620, HCT116, LOVO, A549, H441, H727, LU99 and A427), but It showed no obvious inhibitory effect in wild-type independent cell lines (such as A375) and had good selectivity; it also had significant inhibitory activity on the pERK levels of AsPC-1 and GP2D cells; in various pharmacokinetic experiments, the compounds of the present invention showed good pharmacokinetic properties (such as high exposure and long half-life), and relative to human and mouse plasma, the compounds of the present invention had a higher distribution in whole blood and red blood cells; in in vivo efficacy experiments, the compounds of the present invention showed excellent tumor inhibition effects, with small dosage and high safety, and have broad application prospects.

[0284] Definition and Description

[0285] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

[0286] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0287] The term "pharmaceutically acceptable salt" refers to salts of the compounds of the present invention, prepared from compounds with specific substituents discovered by the present invention and relatively non-toxic acids or bases. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds of the present invention contain basic and acidic functional groups and can be converted into either base or acid addition salts.

[0288] Pharmaceutically acceptable salts of the present invention can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0289] The compounds of the present invention may exist in specific geometric or stereoisomeric forms. The present invention contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present invention.

[0290] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0291] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability of a ring to rotate freely about double bonds or single bonds of ring carbon atoms.

[0292] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0293] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0294] Unless otherwise specified, the key is a solid wedge. and dotted wedge key To indicate the absolute configuration of a stereocenter, use a straight solid bond and straight dashed key To indicate the relative configuration of a stereocenter, use a wavy line Indicates a wedge-shaped solid key and / or dotted wedge key Or use a wavy line Represents a straight solid bond and / or straight dashed key

[0295] Certain compounds of the present invention may exist as atropisomers, which are conformational isomers that occur when rotation about a single bond in a molecule is prevented or greatly slowed due to steric interactions with other parts of the molecule. The compounds disclosed herein include all atropisomers, either pure individual atropisomers, or enriched in one of the atropisomers, or nonspecific mixtures of each. If the rotational potential about a single bond is high enough and the interconversion between conformations is slow enough, separation of the isomers may be permitted.

[0296] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0297] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0298] Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present invention is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (for example, a carbamate is generated from an amine). The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more atoms that constitute the compound. For example, the compound can be labeled with a radioactive isotope, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For another example, deuterium can be used to replace hydrogen to form a deuterated drug. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing efficacy, and extending the biological half-life of drugs. All isotopic composition changes of the compounds of the present invention, whether radioactive or not, are included in the scope of the present invention.

[0299] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include a variant of deuterium and hydrogen, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. The term "optionally substituted" means that it may be substituted or not substituted, and unless otherwise specified, the type and number of the substituent may be arbitrary on the basis of chemical achievable.

[0300] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice. In addition, combinations of substituents and / or variants thereof are permitted only if such combinations result in stable compounds.

[0301] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0302] When the number of a substituent is 0, it means that the substituent does not exist, for example, -A-(R)0 means that the structure is actually -A.

[0303] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A.

[0304] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0305] When the listed linking group does not indicate its linking direction, its linking direction is arbitrary, for example, The connecting group L is -MW-, in which case -MW- can connect ring A and ring B in the same direction as the reading order from left to right to form You can also connect ring A and ring B in the opposite direction of the reading order from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0306] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bonds are connected, the number of H atoms at the site will decrease with the number of connected chemical bonds to become a group with the corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy line For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; It means that any connectable site on the piperidine group can be connected to other groups through one chemical bond, including at least These four connection methods, even if the H atom is drawn on -N-, Still includes For groups connected in this way, when one chemical bond is connected, the H at that site will be reduced by one and become a corresponding monovalent piperidine group.

[0307] Unless otherwise specified, the number of atoms in a ring is generally defined as the ring member number, for example, a "5-7 membered ring" refers to a "ring" having 5-7 atoms arranged around it.

[0308] Unless otherwise specified, the term “C 1-6 "Alkyl" by itself or in combination with other terms is used to refer to a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1-6 Alkyl groups include C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C6 and C5 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6 Examples of alkyl include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, etc. Unless otherwise specified, the term “C 1-4 "Alkyl" by itself or in combination with other terms is used to refer to a straight or branched chain saturated hydrocarbon group consisting of 1 to 4 carbon atoms. 1-4 Alkyl groups include C 1-2 , C 1-3 and C 2-3 Alkyl, etc.; it may be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-4 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), and the like.

[0309] Unless otherwise specified, the term “C 1-4 "Alkoxy" by itself or in combination with other terms refers to those alkyl groups containing 1 to 4 carbon atoms, which are attached to the rest of the molecule through an oxygen atom. 1-4 Alkoxy includes C 1-3 , C 1-2 , C 2-4 , C4 and C3 alkoxy, etc. It can be monovalent, divalent or polyvalent. 1-4 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy (including n-propoxy and isopropoxy), butoxy (including n-butoxy, isobutoxy, s-butoxy and t-butoxy), and the like.

[0310] Unless otherwise specified, “C 3-6"Cycloalkyl" by itself or in combination with other terms refers to a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms. 3-6 The cycloalkyl group includes monocyclic and polycyclic rings, wherein the polycyclic rings include spirocyclic rings, fused rings and bridged rings. 3-6 Cycloalkyl includes C 3-5 , C 4-5 and C 5-6 Cycloalkyl, etc.; it may be monovalent, divalent or polyvalent. 3-6 Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[1.1.1]pentanyl, and the like.

[0311] Unless otherwise specified, the term "3-10 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated or partially unsaturated cyclic group consisting of 3 to 10 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the carbon atoms are optionally oxo (i.e., forming C=O), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). The 3-10 membered heterocycloalkyl includes monocyclic and polycyclic rings, wherein the polycyclic ring includes spirocyclic rings, fused rings and bridged rings. In addition, for the "3-10 membered heterocycloalkyl", heteroatoms can occupy the position where the heterocycloalkyl is connected to the rest of the molecule. The 3-10 membered heterocycloalkyl includes 3-6 membered, 4-6 membered, 5-6 membered, 4-7 membered, 5-7 membered, 5-8 membered, 6-8 membered, 6-9 membered, 6-10 membered, 4 membered, 5 membered, 6 membered, 7 membered, 8 membered, 9 membered and 10 membered heterocycloalkyl, etc. It can be monovalent, divalent or polyvalent. Examples of 3-10 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl or hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, tetrahydropyridinyl, wait.

[0312] Unless otherwise specified, the term "3-7 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated or partially unsaturated cyclic group consisting of 3 to 7 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the carbon atoms are optionally oxo (i.e., forming C=O), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p , p is 1 or 2). The 3-7 membered heterocycloalkyl includes monocyclic and polycyclic rings, wherein the polycyclic ring includes spirocyclic rings, fused rings and bridged rings. In addition, for the "3-7 membered heterocycloalkyl", heteroatoms may occupy the position where the heterocycloalkyl is connected to the rest of the molecule. The 3-7 membered heterocycloalkyl includes 3-6 membered, 4-6 membered, 5-6 membered, 4-7 membered, 5-7 membered, 4 membered, 5 membered, 6 membered and 7 membered heterocycloalkyl, etc. It can be monovalent, divalent or polyvalent. Examples of 3-7 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl or hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, tetrahydropyridinyl, etc.

[0313] Unless otherwise specified, the term "3-6 membered heterocycloalkyl" by itself or in combination with other terms refers to a saturated or partially unsaturated cyclic group consisting of 3 to 6 ring atoms, 1, 2, 3 or 4 of which are heteroatoms independently selected from O, S and N, and the rest are carbon atoms, wherein the carbon atoms are optionally oxo (i.e., forming C=O), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O)). p, p is 1 or 2). The 3-6 membered heterocycloalkyl includes monocyclic and polycyclic rings, wherein the polycyclic ring includes spirocyclic rings, fused rings and bridged rings. In addition, for the "3-6 membered heterocycloalkyl", a heteroatom may occupy the position where the heterocycloalkyl connects to the rest of the molecule. The 3-6 membered heterocycloalkyl includes 4-6 membered, 5-6 membered, 4 membered, 5 membered and 6 membered heterocycloalkyl, etc. It can be monovalent, divalent or polyvalent. Examples of 3-6 membered heterocycloalkyl groups include, but are not limited to, azetidinyl, oxetanyl, thietanyl, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, hexahydropyridazinyl or piperidinyl, etc.

[0314] Unless otherwise specified, the terms "5-6 membered heteroaromatic ring" and "5-6 membered heteroaryl" of the present invention can be used interchangeably. The term "5-6 membered heteroaryl" means a monocyclic group with a conjugated π electron system consisting of 5 to 6 ring atoms, wherein 1, 2, 3 or 4 ring atoms are heteroatoms independently selected from O, S and N, and the rest are carbon atoms. The carbon atoms are optionally oxidized (i.e., forming C=O), the nitrogen atoms are optionally quaternized, and the nitrogen and sulfur heteroatoms are optionally oxidized (i.e., NO and S(O) p , p is 1 or 2). The 5-6 membered heteroaryl can be connected to the rest of the molecule via a heteroatom or a carbon atom. The 5-6 membered heteroaryl includes 5-membered and 6-membered heteroaryl. It can be monovalent, divalent or polyvalent. Examples of the 5-6 membered heteroaryl include, but are not limited to, pyrrolyl (including N-pyrrolyl, 2-pyrrolyl and 3-pyrrolyl, etc.), pyrazolyl (including 2-pyrazolyl and 3-pyrazolyl, etc.), imidazolyl (including N-imidazolyl, 2-imidazolyl, 4-imidazolyl and 5-imidazolyl, etc.), oxazolyl (including 2-oxazolyl, 4-oxazolyl and 5-oxazolyl, etc.), triazolyl (1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, 1H-1,2,4-triazolyl and 4H-1,2,4-triazolyl, etc.), tetrazolyl, isoxazolyl (3-isoxazolyl, 4-isoxazolyl and 5-isoxazolyl, etc.), thiazolyl (including 2-thiazolyl, 4-thiazolyl and 5-thiazolyl, etc.), furanyl (including 2-furanyl and 3-furanyl, etc.), thienyl (including 2-thienyl and 3-thienyl, etc.), pyridyl (including 2-pyridyl, 3-pyridyl and 4-pyridyl, etc.), pyrazinyl or pyrimidinyl (including 2-pyrimidinyl and 4-pyrimidinyl, etc.).

[0315] Unless otherwise specified, the term "5-membered heteroaryl and 5-membered heteroaryl" in the present invention means a 5-membered heteroaryl fused to another 5-membered heteroaryl through two adjacent atoms. Examples of "5-membered heteroaryl and 5-membered heteroaryl" include but are not limited to

[0316] Unless otherwise specified, C n-n+m or C n -C n+m Any specific case including n to n+m carbons, such as C 1-12 Including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 , also includes any range from n to n+m, such as C 1-12 Including C 1-3 , C 1-6 , C 1-9 , C 3-6 , C 3-9 , C 3-12 , C 6-9 , C 6-12 , and C 9-12 Etc.; similarly, n-membered to n+m-membered means that the number of atoms in the ring is n to n+m, for example, 3-12-membered ring includes 3-membered ring, 4-membered ring, 5-membered ring, 6-membered ring, 7-membered ring, 8-membered ring, 9-membered ring, 10-membered ring, 11-membered ring, and 12-membered ring, and also includes any range from n to n+m, for example, 3-12-membered ring includes 3-6-membered ring, 3-9-membered ring, 5-6-membered ring, 5-7-membered ring, 6-7-membered ring, 6-8-membered ring, and 6-10-membered ring, etc.

[0317] The term "leaving group" refers to a functional group or atom that can be replaced by another functional group or atom through a substitution reaction (e.g., a nucleophilic substitution reaction). For example, representative leaving groups include trifluoromethanesulfonate; chlorine, bromine, iodine; sulfonate groups, such as mesylate, tosylate, p-brosylate, p-toluenesulfonate, etc.; acyloxy groups, such as acetoxy, trifluoroacetoxy, etc.

[0318] The term "protecting group" includes, but is not limited to, "amino protecting group", "hydroxy protecting group" or "thiol protecting group". The term "amino protecting group" refers to a protecting group suitable for preventing side reactions at the amino nitrogen position. Representative amino protecting groups include, but are not limited to: formyl; acyl, such as alkanoyl (such as acetyl, trichloroacetyl or trifluoroacetyl); alkoxycarbonyl, such as tert-butyloxycarbonyl (Boc); arylmethoxycarbonyl, such as benzyloxycarbonyl (Cbz) and 9-fluorenylmethoxycarbonyl (Fmoc); arylmethyl, such as benzyl (Bn), trityl (Tr), 1,1-bis-(4'-methoxyphenyl)methyl; silyl, such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), etc. The term "hydroxy protecting group" refers to a protecting group suitable for preventing side reactions of the hydroxyl group. Representative hydroxy protecting groups include, but are not limited to, alkyl groups such as methyl, ethyl and tert-butyl; acyl groups such as alkanoyl (e.g., acetyl); arylmethyl groups such as benzyl (Bn), p-methoxybenzyl (PMB), 9-fluorenylmethyl (Fm) and diphenylmethyl (diphenylmethyl, DPM); silyl groups such as trimethylsilyl (TMS) and tert-butyldimethylsilyl (TBS), and the like.

[0319] The compounds of the present invention can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include but are not limited to the embodiments of the present invention.

[0320] The structure of the compound of the present invention can be confirmed by conventional methods known to those skilled in the art. If the present invention relates to the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data of the cultured single crystal using a Bruker D8 venture diffractometer, the light source is CuKα radiation, and the scanning mode is: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0321] The solvents used in the present invention can be obtained from commercial sources. The software names were used, and commercially available compounds were named using the supplier's catalog names. BRIEF DESCRIPTION OF THE DRAWINGS

[0322] Figure 1 Diagram of the binding pattern of compound A and CypA protein.

[0323] Figure 2 Binding pattern of compound B and CypA protein.

[0324] Figure 3 Binding pattern of compound C and CypA protein.

[0325] Figure 4 Binding pattern of compound D and CypA protein.

[0326] Figure 5 Binding pattern diagram of compound E and CypA protein.

[0327] Figure 6 Binding pattern of compound F and CypA protein.

[0328] Figure 7 Binding pattern of compound G and CypA protein.

[0329] Figure 8 Binding pattern of compound H to CypA protein.

[0330] Fig. 9 Binding pattern diagram of compound I and CypA protein.

[0331] Fig.10 Binding pattern of compound J and CypA protein. DETAILED DESCRIPTION

[0332] The present invention is described in detail below by way of examples, but it is not intended to impose any adverse limitations on the present invention. The present invention has been described in detail herein, and specific embodiments thereof are also disclosed therein. It will be apparent to those skilled in the art that various changes and modifications may be made to the specific embodiments of the present invention without departing from the spirit and scope of the present invention.

[0333] Calculation Example 1

[0334]

[0335] Binding mode prediction used the co-crystal complex of human CypA protein and natural product Sanglifehrin A (PDB ID code: 1YND) as docking template. To prepare the protein, Maestro [1] The protein preparation wizard module was used to add hydrogen atoms and the OPLS4 force field was used to optimize the hydrogen bonds of the cocrystal structure and remove the ligand Sanglifehrin A in the cocrystal complex. For ligand preparation: generate a 3D structure of the molecule to be docked and perform energy minimization using LigPrep [2] Using Maestro( Induced Fit Docking in version 2021-2 [3] Using the Protocol: Extended Sampling option, dock compound A into the prepared 1YND protein structure. Select the best binding model, see Figure 1 The selected model retains the main hydrogen bonding between the ligand Sanglifehrin A and the protein in the original cocrystal complex. The centroid of compound A in this binding model is used to generate Docking the mesh with Glide [4] The Receptor Grid Generation module generates a docking model. Based on this docking model, compounds B to J were docked using Glide [3] The SP docking mode in the figure is used for docking. The binding modes of compounds B to J are shown in Figure 2 to Figure 10 .

[0336] [1] Maestro, LLC,New York,NY,2021.

[0337] [2]LigPrep, LLC,New York,NY,2021.

[0338] [3]Induced Fit Docking protocol; Glide, LLC,New York,NY,2021;Prime, LLC,New York,NY,2021.

[0339] [4] Glide, LLC,New York,NY,2021.

[0340] Conclusion: The compounds of the present invention have good binding with human Cyp A protein. The compounds of the present invention form hydrogen bonds with Arg55, Gln63, Asn102 and His126. In addition, Arg55 forms a cationic π bond with the indole ring. Since the compounds of the present invention act on the protein surface, the listed hydrogen bonds as anchor points not only reproduce the binding mode of the natural product Sanglifehrin A in the co-crystal complex, but also tightly fix the compounds of the present invention on the protein surface. The binding of the compounds of the present invention to CypA will block the binding of RAF downstream of RAS to RAS, thereby inhibiting the RAS-RAF-MEK-ERK signaling pathway to achieve anti-tumor effects.

[0341] Reference Example 1: Compound M1

[0342]

[0343] Compound M1-1 (39 g, 106.78 mmol) was dissolved in water (100 mL) and tetrahydrofuran (200 mL), and lithium hydroxide monohydrate (13.44 g, 320.33 mmol) was added. The reaction solution was stirred at 25°C for 1.5 hours. After the reaction was completed, the pH of the reaction solution was adjusted to 5-6 with 2M dilute hydrochloric acid, and then the reaction solution was extracted with ethyl acetate (200 mL*3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and the organic solvent was removed by distillation under reduced pressure to obtain compound M1. LCMS: m / z=372.8,374.8[M+23] + .

[0344] Reference Example 2: Compound M2

[0345]

[0346]

[0347] Step 1

[0348] Compound M2-1 (150 g, 1.14 mol) was dissolved in pyridine (200 mL), and then 4-dimethylaminopyridine (7.4 g, 60.57 mmol) and acrolein (38.58 g, 688.11 mmol) were added. The reaction solution was stirred at 50 °C for 48 hours. The reaction solution was cooled to room temperature, then poured into 2000 mL of water, extracted with ethyl acetate (300 mL*3), the combined organic phases were washed with dilute hydrochloric acid (2M, 300 mL*3), then washed once with water (500 mL), and then washed once with saturated brine (200 mL), dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product. The crude product was then distilled under reduced pressure at about -0.1 MPa and 90-95 °C to obtain compound M2-2.

[0349] Step 2

[0350] Compound M2-2 (17 g, 134.76 mmol) and di-tert-butyl azodicarboxylate (31.03 g, 134.76 mmol) were dissolved in toluene (170 mL), and the reaction solution was stirred at 80° C. for 16 hours. The reaction solution was concentrated, and the crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-15%) to obtain compound M2-3. 1H NMR (400MHz, CDCl3) δppm 5.93 (s, 2H), 5.06-5.37 (m, 1H), 4.13-4.24 (m, 3H), 3.57-3.79 (m, 1H), 1.48 (s, 18H), 1.27-1.30 (m, 3H).

[0351] Step 3

[0352] Compound M2-3 was subjected to SFC chiral separation (column: DAICEL CHIRALPAK IC (250mm*50mm, 10um); mobile phase: phase A supercritical carbon dioxide, phase B [0.1% ammonia in isopropanol]; B%: 20%-20%) to obtain compound M2-3A. SFC analysis method (column: Cellulose 2 (150mm*4.6mm, ID, 5um; mobile phase: phase A supercritical carbon dioxide, phase B [0.05% diethylamine in isopropanol]; isocratic elution B%: 5%-5%, column temperature: 35°C, column pressure: 1500psi), ee=100%, the elution time of compound M2-3A was 2.798min, and the elution time of its enantiomer was 2.133min. LCMS: m / z=379.0[M+23]+.

[0353] Step 4

[0354] Under nitrogen protection, trimethyl sulfoxide iodide (3.09g, 14.03mmol) was dissolved in dimethyl sulfoxide (10mL), and then potassium tert-butoxide (1.26g, 11.22mmol) was added, and the mixture was stirred at 50°C for 2 hours. Compound M2-3A (1g, 2.81mmol) was then added, and the reaction solution was stirred at 70°C for 12 hours. Water (100mL) was added to dilute the reaction solution, and the solution was extracted with ethyl acetate (50mL*3). The organic phases were combined and washed once with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-15%) to obtain compound M2-5. LCMS: m / z=371.2[M+1] + ; 1 H NMR (400MHz, CDCl3) δppm 4.22-4.33(m,1H),4.07-4.20(m,2H),2.56-2.73(m,1H),2.11-2.24(m,1H),1.84-1.9 5(m,2H),1.71-1.82(m,1H),1.45-1.53(m,18H),1.23-1.31(m,3H),0.97-1.07(m,1H).

[0355] Step 5

[0356] Compound M2-5 (630.00 mg, 1.70 mmol) was dissolved in dichloromethane (2 mL), and then trifluoroacetic acid (3.07 g, 26.93 mmol, 2 mL) was added. The reaction solution was stirred at 20°C for 2 hours. The reaction solution was concentrated to obtain the crude trifluoroacetate salt of M2-6, which was used directly in the next step.

[0357] Step 6

[0358] The crude trifluoroacetate of compound M2-6 obtained in step 5 was dissolved in dichloromethane (10 mL), and then N-methylmorpholine (959.97 mg, 9.49 mmol), compound M1 (0.4 g, 1.14 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (364 mg, 1.90 mmol), and 1-hydroxybenzotriazole (39 mg, 284.8 μmol) were added. The reaction solution was stirred at 25 ° C for 12 hours. Water (50 mL) was added to dilute the reaction solution, and the solution was extracted with dichloromethane (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-60%) to obtain compound M2. LCMS: m / z = 503.1, 505.1 [M+1] + .

[0359] Reference Example 3: Compound M3

[0360]

[0361] Step 1

[0362] Tetrabutylammonium bromide (116.14 mg, 360.26 μmol) and difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol) were added to a toluene (24 mL) solution of compound M2-3A (4.28 g, 12.01 mmol). The mixture was stirred at 110°C for 4 hours. Difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol) was added and stirred at 110°C for 16 hours. Difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol) was added and stirred at 110°C for 4 hours. Difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol) was added again and stirred at 110°C for 16 hours. The toluene was removed by concentration under reduced pressure, 20 mL of water was added to the residue, and the mixture was extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (10 mL*1), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by rapid silica gel column chromatography (0-10% ethyl acetate / petroleum ether) to obtain compound M3-1. LCMS: m / z=206.9[M-2Boc+1] + ; 1 H NMR (400MHz, CDCl3) δ4.96-5.25(m,1H),4.17-4.51(m,3H),3.19(br s,1H),2.18(br d,J=10.54Hz,1H),2.00-2.11(m,1H),1.40-1.55(m,18H),1.30-1.38(m,3H).

[0363] Step 2

[0364] Compound M3-1 (1 g, 2.46 mmol) was dissolved in dichloromethane (10 mL), trifluoroacetic acid (7.68 g, 67.31 mmol, 5 mL) was added, and the reaction solution was stirred at 25°C for 3 hours. After the reaction was completed, the reaction solution was distilled under reduced pressure to remove the organic solvent to obtain the crude trifluoroacetate salt of compound M3-2, which was used in the next step. LCMS: m / z = 206.8 [M+1] + .

[0365] Step 3

[0366] At 0°C, compound M1 (1.04 g, 2.95 mmol) and compound M3-2 (507 mg, crude trifluoroacetate) were dissolved in dichloromethane (10 mL), and N-methylmorpholine (2.49 g, 24.59 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (1.41 g, 7.38 mmol), and 1-hydroxybenzotriazole (66.45 mg, 491.78 μmol) were added in sequence. The reaction solution was stirred at 25°C for 3 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (20-60% ethyl acetate / petroleum ether) to obtain compound M3. LCMS: m / z=539.1,541.1[M+1] + .

[0367] Reference Example 4: Compound M4

[0368]

[0369] Step 1

[0370] At 0°C, under nitrogen protection, a tetrahydrofuran solution of compound M4-1 (25g, 136.61mmol) was added to a tetrahydrofuran solution of methyl magnesium bromide (3M, 91.07mL), and the mixture was stirred at 0°C for two hours. The reaction solution was slowly poured into ice water, and concentrated hydrochloric acid was added to adjust the pH to 6-7. Ethyl acetate was added for extraction (500mL*2), the organic phases were combined and then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was chromatographed on a rapid silica gel column (ethyl acetate / petroleum ether 0-30%) to obtain M4-2. LCMS: m / z=200.0, 202.0[M+1] + ; 1 H NMR (400MHz, CDCl3) δ8.58 (d, J = 3.2 Hz, 1H), 8.00 (d, J = 8.4 Hz, 1H), 7.29 (dd, J = 8.4, 4.8 Hz, 1H), 2.69 (s, 3H).

[0371] Step 2

[0372] At 0°C, under nitrogen protection, compound (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethanediamine (p-isopropylbenzene) ruthenium chloride (700.83mg, 1.10mmol) was added to a mixed solvent of formic acid (12.68g, 263.96mmol) and triethylamine (133.55g, 1.32mol, 183.70mL). Then, it was stirred at 40°C for 15 minutes, then cooled to 25°C, M4-2 (22g, 109.98mmol) was added, and then stirred at 40°C for 2 hours. The reaction solution was concentrated under reduced pressure, the residue was diluted with 100mL, extracted with ethyl acetate (50mL*2), the organic phases were combined and then dried with anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product. The crude product was chromatographed on a rapid silica gel column (ethyl acetate / petroleum ether 0-40%) to obtain compound M4-3. LCMS: m / z = 202.0, 204.0 [M+1] + ; 1 H NMR (400MHz, CDCl3) δ8.50(d,J=4.8Hz,1H),7.84(d,J=8.0Hz,1H),7.13(dd,J=8.0,4.8Hz,1H),5.11(br s,1H),4.44(br s,1H)1.44(d,J=4.8Hz,3H).

[0373] Step 3

[0374] At 0°C, under nitrogen protection, sodium hydrogen (5.23 g, 130.66 mmol, 60% purity) was added in batches to a solution of compound M4-3 (22 g, 108.88 mmol) in tetrahydrofuran (200 mL). Stir at 0°C for 1 hour, add iodomethane (217.77 mmol, 13.56 mL), and after the addition, slowly heat the mixture from 0°C to 25°C and stir at this temperature for 2 hours. Add saturated aqueous ammonium chloride solution (100 mL) to the mixture at 0°C to quench, add water (200 mL), and then extract with ethyl acetate (100 mL*2), collect and combine the organic phases, add anhydrous sodium sulfate, dry and filter, and concentrate the filtrate under reduced pressure. The crude product is separated and purified by silica gel column (ethyl acetate: petroleum ether 0-20%) to obtain compound M4-4. LCMS: m / z=216.0,218.0[M+1] + ; 1 HNMR (400MHz, CDCl3) δ8.59 (d, J=3.2Hz, 1H), 7.82 (dd, J=8.4, 3.2Hz, 1H), 7.08 (dd, J=8.0, 4.4Hz, 1H), 4.91 (dd, J=13.2, 6.4Hz, 1H), 3.29 (s, 3H), 1.44 (d, J= 4.8Hz, 3H).

[0375] Step 4

[0376] To a toluene (50 mL) solution of compound M4-4 (5 g, 23.14 mmol), bis-pinacol borate (7.05 g, 27.77 mmol), Pd(dppf)Cl2 (1.69 g, 2.31 mmol) and potassium acetate (4.54 g, 46.28 mmol) were added. After the addition, nitrogen was replaced three times. Stir for 3 hours at 100°C. Cool to room temperature, add water (100 mL), then extract with ethyl acetate (50 mL*2), collect and combine the organic phases, add anhydrous sodium sulfate to dry, filter, and concentrate the filtrate under reduced pressure. The crude product was separated and purified by silica gel column (ethyl acetate: petroleum ether 0-25%) to obtain compound M4. LCMS: m / z=182.0 [boric acid M+1] + ; 1 H NMR (400MHz, CDCl3) δ8.59 (dd, J=4.8, 1.6Hz, 1H), 7.90 (dd, J=7.6, 2.0Hz, 1H), 7.16 (dd, J=8.4,4.8Hz,1H),4.77(dd,J=13.2,6.4Hz,1H),3.61(s,3H),3.26(s,3H)1.37(s,12H).

[0377] Reference Example 5: Compound M5

[0378]

[0379] Step 1

[0380] Compound M5-1 (18.3 g, 138.47 mmol), tert-butyldiphenylsilyl chloride (40 g, 145.53 mmol) and imidazole (12.3 g, 180.68 mmol) were dissolved in 300 ml of anhydrous water, and the mixture was stirred at 15°C for 20 hours. After concentration, the crude product was diluted with 200 ml of water, extracted with ethyl acetate (3*200 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-10%) to obtain compound M5-2. 1 H NMR (400MHz, CDCl3) δ = 7.67 (dd, J = 1.6, 8.0Hz, 4H), 7.49-7.36 (m, 6H), 3.70 (s, 3H), 3.66 (s, 2H), 1.22 (s, 6H), 1.05 (s, 9H).

[0381] Step 2

[0382] Compound M5-2 (51 g, 137.63 mmol) and potassium hydroxide (16.51 g, 294.35 mmol) were dissolved in 200 ml of water and 200 ml of ethanol, and the mixture was stirred at 90°C for 3 hours. The reaction solution was concentrated to about 250 ml, diluted with 500 ml of ethyl acetate, and the pH was adjusted to 3-4 with concentrated hydrochloric acid, and the liquids were separated. The aqueous phase was extracted with ethyl acetate (2*500 mL), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound M5-3. 1 H NMR (400MHz, CDCl3) δ = 7.58 (dd, J = 1.6, 8.0Hz, 4H), 7.39-7.27 (m, 6H), 3.58 (s, 2H), 1.15 (s, 6H), 0.97 (s, 9H).

[0383] Step 3

[0384] At 0°C, oxalyl chloride (233.64 mmol, 20.45 mL) was slowly added dropwise to a solution of compound M5-3 (49 g, 137.44 mmol) and DMF (13.74 mmol, 1.06 mL) in 500 mL of anhydrous dichloromethane, and the mixture was stirred at 20°C for 15 hours. The reaction solution was concentrated, 300 mL of anhydrous toluene was added, and the mixture was concentrated under reduced pressure to obtain compound M5-4.

[0385] Step 4

[0386] At 0°C, tin tetrachloride (136.00mmol, 15.92mL) was slowly added to a 200mL anhydrous dichloromethane solution of compound M5-4 (51.00g, 136.00mmol), and the mixture was stirred for half an hour under a nitrogen atmosphere. At 0°C, 200mL anhydrous dichloromethane of compound M5-5 (26.66g, 136mmol) was slowly added dropwise to the above reaction mixture, and the reaction system was stirred for 1 hour at 0°C under a nitrogen atmosphere. 500mL of water was added to quench, filtered, and the filtrate was allowed to stand for separation. The aqueous phase was extracted with dichloromethane (3*500mL), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-30%) to obtain compound M5-6. LCMS: m / z=534.1,536.1[M+1] + ; 1H NMR (400MHz, CDCl3) δ = 8.70 (d, J = 1.6Hz, 1H), 8.62 (br s,1H),7.69(d,J=3.2Hz,1H),7.57-7.51(m,4H),7.44-7.39(m,2H),7.38-7.27(m,6H),3.96-3.88(m,2H),1.44(s,6H),0.98(s,9H).

[0387] Step 5

[0388] At 0°C, under nitrogen atmosphere, slowly add lithium borohydride tetrahydrofuran solution (1M, 123.47mL) to 220mL of anhydrous tetrahydrofuran of compound M5-6 (22g, 41.16mmol). After the addition, heat to 60°C and stir for 15 hours. Cool to room temperature, slowly add 10mL of saturated ammonium chloride aqueous solution, then add 100mL of ethyl acetate, wash with 50mL of saturated brine, collect the organic phase, dry with anhydrous sodium sulfate, filter, and concentrate the filtrate. The residue is purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-20%) to obtain compound M5-7. LCMS: m / z=520.1,522.1[M+1] + .

[0389] Step 6

[0390] Compound M5-7 (18.5 g, 35.54 mmol), elemental iodine (9.02 g, 35.54 mmol) and silver trifluoromethanesulfonate (10.04 g, 39.09 mmol) were added to 185 ml of anhydrous tetrahydrofuran in sequence, and the mixture was stirred at 20°C for 2 hours. 50 ml of saturated sodium sulfite aqueous solution was added to quench, 200 ml of ethyl acetate was added to dilute, and the mixture was filtered. The filtrate was allowed to stand for separation, and the organic phase was collected and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound M5. 1 H NMR (400MHz, CDCl3) δ = 8.07 (s, 1H), 7.78-7.69 (m, 5H), 7.49-7.40 (m, 6H), 7.24-7. 20(m,1H),7.19-7.16(m,1H),3.50(s,2H),2.71(s,2H),1.17(s,9H),0.95(s,6H).

[0391] Reference Example 6: Compound M6

[0392]

[0393] Step 1

[0394] Compound M6-1 (10 g, 78.05 mmol) was dissolved in DCM (100 mL) and MeOH (20 mL), and trimethylsilyldiazomethane hexane solution (2 M, 78.05 mL) was added dropwise at 0° C. Stirring was continued at 0° C. for 10 minutes. Concentration gave compound M6-2, which was used directly in the next step.

[0395] Step 2

[0396] Compound M6-2 (10 g, 70.35 mmol) was dissolved in n-heptane (200 mL), tert-butyl carbazate (11.69 g, 70.35 mmol) was added, and then the temperature was raised to 70°C and stirred for 12 hours. Stirring was stopped, and when the system was cooled to room temperature, water (100 mL) and ethyl acetate (100 mL) were added, and the separated organic phase was dried with anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound M6-3, which was directly used in the next step. LCMS: m / z=201[M+1-56] + .

[0397] Step 30°C, under nitrogen protection, slowly drop a solution of compound M6-3 (7g, 27.31mmol) in tetrahydrofuran (10mL) into borane dimethyl sulfide (10M, 273.12mL), stir at 0°C for half an hour, then warm to room temperature and continue stirring for 1 hour. Slowly drop methanol (250mL) at 0°C to quench the reaction, and concentrate the quenched reaction solution under reduced pressure. The crude product is purified by column chromatography (eluent: ethyl acetate / petroleum ether 3% to 7%) to obtain compound M6-4. LCMS: m / z=203[M+1-56] + .

[0398] Step 4

[0399] Compound M6-4 (1.7 g, 6.58 mmol) was dissolved in tetrahydrofuran (30 mL), and di-tert-butyl dicarbonate (2.15 g, 9.87 mmol), triethylamine (2.00 g, 19.74 mmol) and 4-dimethylaminopyridine (80.40 mg, 658.12 μmol) were added in sequence at 25°C, and stirred at 25°C for 1 hour. Water (50 mL) and ethyl acetate (30 mL) were added to the reaction system, and the organic phase obtained by extraction was dried over anhydrous sodium sulfate, and the filtrate was concentrated. The crude product was purified by column chromatography (eluent: ethyl acetate / petroleum ether 3% to 7%) to obtain compound M6-5. LCMS: m / z=359[M+1] + .

[0400] Step 5

[0401] Compound M6-5 (585 mg, 1.63 mmol) was dissolved in tetrahydrofuran (20 mL). Under nitrogen protection, bis(trimethylsilyl) lithium amide (1 M, 4.90 mL) was added dropwise at -70 °C. After stirring at -70 °C for half an hour, trimethylsilyl chloride (531.94 mg, 4.90 mmol) was added dropwise. After stirring at -70 °C for one hour, N-bromosuccinimide (1.16 g, 6.53 mmol) was added. The temperature was slowly raised to 25 °C and stirring was continued for 1 hour. Saturated brine (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound M6-6, which was used directly in the next step. LCMS: m / z=353,355[M+1-100-56] + .

[0402] Step 6

[0403] Add a solution of citric acid (618.66 mg, 2.94 mmol) in water (5 mL) to a solution of compound M6-6 (500 mg, 981.34 μmol) in tetrahydrofuran (20 mL) and stir at 25°C for 1 hour. After the reaction is complete, add saturated brine (30 mL) and extract with ethyl acetate (30 mL). The organic phase is dried over anhydrous sodium sulfate and filtered. After the filtrate is concentrated, it is purified by column chromatography (eluent: ethyl acetate / petroleum ether 3% to 7%) to obtain compound M6-7. LCMS: m / z=281,283[M+1-100-56] + .

[0404] Step 7

[0405] Compound M6-7 (150 mg, 342.99 μmol) was dissolved in acetonitrile (20 mL), cesium carbonate (447.02 mg, 1.37 mmol) was added, and the mixture was heated to 60°C and stirred for 12 hours. The mixture was filtered, and the filtrate was concentrated and purified by column chromatography (eluent: ethyl acetate / petroleum ether 3% to 7%) to obtain compound M6-8. LCMS: m / z = 201 [M+1-100-56] + . 1 H NMR(400MHz, CDCl3)5.24-4.93(m,1H),4.45(br d,J=4.6Hz,1H),3.77-3.71(m,3H),2.89(qd,J=5.8,11.6Hz,1H),2.39(td,J=4 .9,10.0Hz,1H),2.13(td,J=5.0,10.0Hz,1H),1.62(s,2H),1.52-1.48(m,18H).

[0406] Step 8

[0407] Compound M6-8 (50 mg, 140.29 μmol) was dissolved in methanol (5 mL) and ethyl acetate (5 mL), and a hydrogen chloride ethyl acetate solution (5 mL, 4 M) was added, and stirred at 40°C for 1 hour. The reaction solution was concentrated to obtain the hydrochloride salt of compound M6. LCMS: m / z = 157 [M+1] + .

[0408] Reference Example 7: Compound M8

[0409]

[0410] Step 1

[0411] Triethylamine (111.94mmol, 15.58mL) and 1-methylpiperazine (16.82g, 167.91mmol, 18.63mL) were added to a tetrahydrofuran solution (250mL) of M8-1 (22.5g, 111.94mmol), and the mixture was stirred at 60°C for 16 hours. The reaction solution was diluted with water (200mL), the solution was extracted with ethyl acetate (100mL*3), the organic phases were combined and washed once with saturated brine (100mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was added with petroleum ether (100mL), filtered, and the filter cake was dried to obtain compound M8-2. 1 H NMR (400MHz, DMSO-d6) δppm 8.40 (d, J = 2.4Hz, 1H), 7.65 (d, J = 2.4Hz, 1H), 3.42-3.48 (m, 4H), 2.36-2.44 (m, 4H), 2.21 (s, 3H).

[0412] Step 2

[0413] Compound M8-2 (10 g, 35.57 mmol) was dissolved in tetrahydrofuran (150 mL), and methylmagnesium chloride (3M tetrahydrofuran solution, 23.7 mL) was added under nitrogen protection at 0°C. The reaction solution was stirred at 0°C for 2 hours. Saturated ammonium chloride (100 mL) was poured into the reaction solution to quench, and the solution was extracted with ethyl acetate (100 mL*3). The organic phases were combined and washed once with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent methanol / dichloromethane, methanol ratio: 0-10%) to obtain compound M8-3. LCMS: m / z=298.1, 300.1[M+1] + . 1HNMR (400MHz, DMSO-d6) δppm 8.38 (d, J = 2.4Hz, 1H), 7.53 (d, J = 2.4Hz, 1H), 3.39-3.47 (m, 4H), 2.54 (s, 3H), 2.40-2.45 (m, 4H), 2.22 (s, 3H).

[0414] Step 3

[0415] Under nitrogen protection at 0°C, triethylamine (57.1g, 563.9mmol) was added dropwise to formic acid (5.27g, 109.67mmol), and then (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethanediamine (p-isopropylbenzene) ruthenium chloride (140mg, 220μmol) was added. The mixture was stirred at 40°C for 15 minutes. Then it was cooled to room temperature and compound M8-3 (6.54g, 21.93mmol) was added in batches. The reaction solution was heated to 50°C and stirred for 12 hours. The reaction solution was directly concentrated, and the crude product was purified by flash chromatography (silica gel, eluent methanol / dichloromethane, methanol ratio: 0-10%) to obtain compound M8-4. LCMS: m / z=300.0, 302.0 [M+1] + .

[0416] Step 4

[0417] Under nitrogen protection, compound M8-4 (1g, 3.33mmol) was dissolved in N,N-dimethylformamide (10mL), cooled to 0℃, and then sodium hydrogen (160mg, 4.00mmol, purity 60%) was added in batches. The solution was stirred at 0℃ for 1 hour, and then iodomethane (520mg, 3.68mmol) was added dropwise, and the reaction solution continued to react at 0℃ for 2 hours. Saturated ammonium chloride (50mL) was added dropwise to the reaction solution to quench, and the solution was extracted with ethyl acetate (50mL*3). The organic phases were combined and washed once with saturated brine (50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated to obtain a crude product, which was purified by flash chromatography (silica gel, eluent methanol / dichloromethane, methanol ratio: 0-10%) to obtain compound M8-5. LCMS: m / z=313.9, 316.0[M+1] + .

[0418] Step 5

[0419] Compound M8-5 (0.72 g, 2.29 mmol) and neopentyl glycol diboronate (777 mg, 3.5 mmol) were dissolved in toluene (20 mL), potassium acetate (563 mg, 5.75 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (168 mg, 230 μmol) were added in sequence, nitrogen was replaced three times, and the mixture was heated to 70°C for 12 hours. The reaction solution was filtered, and the filtrate was directly concentrated. The obtained crude product M8-6 was directly used in the next step. LCMS: m / z=348.1[M+1] + .

[0420] Step 6

[0421] Compound M8-6 (11.05 g, 31.82 mmol,) and compound M5 (24.69 g, 38.18 mmol) were dissolved in dioxane (100 mL) and water (20 mL), potassium carbonate (13.19 g, 95.46 mmol) was added, nitrogen was replaced three times, 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (22.07 mg, 33.86 μmol) was added, and the reaction solution was stirred under nitrogen at 70°C for 12 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (0-15% methanol / dichloromethane) to obtain compound M8-7. LCMS: m / z=753.2,755.2[M+1] + .

[0422] Step 7

[0423] Compound M8-7 (15 g, 19.90 mmol) was dissolved in tetrahydrofuran (150 mL), sodium hydrogen (4 g, 100.00 mmol, purity 60%) was added in batches at 0°C, stirred for 30 min under nitrogen protection, and then iodoethane (30.01 mmol, 2.4 mL) was added dropwise, the reaction solution was stirred at 0°C for 1 hour, iodoethane (25.01 mmol, 2 mL) was added, and the reaction solution was stirred at 20°C for 30 min. The reaction solution was quenched in ice water (100 mL) in batches, extracted with dichloromethane (50 mL*3), the organic phase was washed with saturated brine (50 mL*3), dried over anhydrous sodium sulfate, and the filtrate was concentrated to obtain the crude compound M8-8. LCMS: m / z=781.3,783.3[M+1] + .

[0424] Step 8

[0425] Compound M8-8 (13.7 g, 17.52 mmol) was dissolved in tetrahydrofuran (150 mL), and tetrabutylammonium fluoride tetrahydrofuran solution (1 M, 175.21 mL) was added, and the reaction solution was stirred at 50°C under nitrogen for 12 hours. The reaction solution was quenched with water (20 mL), extracted with ethyl acetate (50 mL*3), the aqueous phase was extracted with dichloromethane (50 mL*3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The residue was purified by column chromatography (0-15% methanol / dichloromethane) to obtain a crude product, and acetonitrile (10 mL) was added to the crude product and stirred for 10 min, filtered, and the filter cake was purified by prep-HPLC (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: the proportion of acetonitrile in the mobile phase increased from 13% to 43% within 8 min) to obtain the trifluoroacetate salt of compound M8-9A (HPLC analysis method: column ChromCore120 C18 3 μm 3.0*30mm; Phase A is a 4-liter aqueous solution containing 1.5 mL of trifluoroacetic acid, and Phase B is a 4-liter acetonitrile solution containing 0.75 mL of trifluoroacetic acid; elution gradient: Phase B increases from 10% to 80% within 6 minutes, then maintains at 80% for 0.5 minutes, and then maintains at 10% for 0.5 minutes; the retention time of compound M8-9A is 3.205 min, and the retention time of its isomer is 3.146 min). LCMS: m / z=543.5,545.5[M+1] + .

[0426] Step 9

[0427] Compound M8-9A (0.6 g, 1.10 mmol) and bis-naphthalene borate (420.47 mg, 1.66 mmol) were dissolved in toluene (6 mL) and dioxane (2 mL), and potassium acetate (216.67 mg, 2.21 mmol) and [1,1-bis(diphenylphosphino)ferrocene]palladium dichloride (80.77 mg, 110.39 μmol) were added. The reaction solution was stirred under nitrogen at 90°C for 5 hours. The reaction solution was filtered, the filtrate was concentrated under reduced pressure, and the crude product was purified by column chromatography (0-15% methanol / dichloromethane) to obtain compound M8. LCMS: m / z=591.4[M+1] + .

[0428] Reference Example 8: Compound M9

[0429]

[0430] Step 1

[0431] Compound M6-1 (78.5 g, 612.68 mmol), compound M9-2 (119.42 g, 673.95 mmol), 4-dimethylaminopyridine (7.49 g, 61.27 mmol) and triethylamine (185.99 g, 1.84 mol) were added to 1.5 liters of anhydrous dichloromethane in sequence, and then 2-chloro-1-methylpyridine (salt) iodide (266.10 g, 1.04 mol) was added in batches, and the mixture was stirred at 25°C for 1 hour. After the reaction was completed, the organic phase was washed with water (2*1 L). The organic phase was dried over sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound M9-3. 1 H NMR (400MHz, CDCl3) δ = 7.40-7.31 (m, 3H), 7.25-7.19 (m, 2H), 4.79-4.66 (m, 1H), 4.29-4.20 (m, 2H), 3.43-3.26 (m, 5H), 3.01-2.79 (m, 4H).

[0432] Step 2

[0433] Compound M9-3 (220 g, 765.72 mmol) and acetic acid (91.97 g, 1.53 mol) were added to 2 liters of anhydrous tetrahydrofuran in sequence. Sodium borohydride (23.18 g, 612.58 mmol) was slowly added to the above solution in batches at 0°C. After the addition was completed, the mixed solution was stirred for 2 hours at 0°C. After the reaction was completed, 500 ml of saturated ammonium chloride aqueous solution was slowly added dropwise, and the solution was concentrated under reduced pressure to about 1 L of residue, and extracted with ethyl acetate (3*500 mL). The organic phase was washed with saturated sodium bicarbonate to pH~8, and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound M9-4. LCMS: m / z=290.1[M+1] + .

[0434] Step 4-dimethylaminopyridine (74.32 g, 608.31 mmol), compound M9-4 (220 g, 760.39 mmol) and N, N-diisopropylethylamine (147.41 g, 1.14 mol) were added to 2 liters of anhydrous dichloromethane at 30°C, and then p-toluenesulfonyl chloride (159.46 g, 836.43 mmol) was added to the above solution in batches. After the addition was completed, the mixture was stirred at 25°C for 3 hours. After the reaction was completed, the mixture was washed with water (1.5 L), the aqueous phase was extracted with dichloromethane (2*500 mL), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-30%) to obtain compound M9-5. 1H NMR (400MHz, CDCl3) δ = 7.81 (br d, J = 8.0Hz, 2H), 7.41-7.28 (m, 5H), 7.19 (br d, J = 7.2Hz, 2H), 4.84-4.70 (m, 1H), 4.65 (br s,1H),4.29-4.18(m,2H),3.26(br d,J=13.6Hz,1H),3.17-2.96(m,2H),2.85-2.71(m,1H),2.60-2.49(m,2H),2.47(s,3H),2.37-2.20(m,1H),1.90(br s,2H).

[0435] Step 4

[0436] Compound M9-5 (90 g, 202.93 mmol) and lithium bromide (35.25 g, 405.85 mmol) were added to 900 ml of 1-methyl-2-pyrrolidone in sequence, and the mixture was stirred at 90°C for 13 hours. After the reaction was completed, 2 liters of saturated brine was added for dilution, and ethyl acetate (3*1 L) was used for extraction. The organic phase was washed again with saturated brine (2*1 L), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-20%) to obtain compound M9-6. LCMS: m / z=352.0, 354.0 [M+1] + .

[0437] Step 5

[0438] At -78°C, compound M9-6 (9.5 g, 23.80 mmol) was dissolved in 95 ml of anhydrous tetrahydrofuran, and then a 2M tetrahydrofuran-n-heptane mixed solution of lithium diisopropylamine (15.47 mL, 30.94 mmol) was slowly added dropwise to stir the mixture for half an hour under a nitrogen atmosphere. A 20 ml anhydrous dichloromethane solution of di-tert-butyl azodicarboxylate (6.58 g, 28.56 mmol) was added to the above solution at one time, and stirring was continued for half an hour. 1,3-dimethyl-tetrahydro-2-pyrimidone (91.50 g, 713.89 mmol) was slowly added to the above reaction solution, and the temperature was naturally raised to room temperature, and stirring was continued for 13 hours. After the reaction was completed, 100 ml of water was slowly added to extract, and lithium hydroxide monohydrate (3.00 g, 71.39 mmol) was added, and stirring was continued at room temperature for 1 hour. Concentrate, add 200 ml of saturated brine to dilute the residue, wash with ethyl acetate (3*200 mL), discard the organic phase, adjust the pH of the aqueous phase to 5 with 1N hydrochloric acid, extract with ethyl acetate (3*200 mL), wash the organic phase with saturated brine (2*200 mL), dry the organic phase with anhydrous sodium sulfate, filter, concentrate, and purify the residue with flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-30%) to obtain compound M9-7. LCMS: m / z=365.1[M+23] + .

[0439] Step 6

[0440] At 25°C, slowly add trimethylsilyldiazomethane 2M n-hexane solution (11.68 mL, 23.36 mmol) to a solution of compound M9-7 (1.6 g, 4.67 mmol) in 32 mL of anhydrous methanol, and stir the mixture at 25°C for 10 minutes. After the reaction is completed, add 0.1 mL of acetic acid to quench the reaction. The reaction solution is concentrated to obtain compound M9-8. LCMS: m / z=379.1[M+23] + . The SFC analysis method was used for detection (column: Cellulose-4 (100mm*4.6mm, 3μm; mobile phase: phase A supercritical carbon dioxide, phase B [0.05% diethylamine in isopropanol]; B%: from 5% to 40% in 4 minutes, then maintained at 40% for 0.5 minutes, and then maintained at 5% for 1.5 minutes) The retention time of compound M9-8 was 1.342min, and the chiral purity was 93.38%; the retention time of its enantiomer was 1.431min, and the chiral purity was 6.62%.

[0441] Step 7

[0442] Compound M9-8 (1.6 g, 4.49 mmol) was dissolved in 5 ml of trifluoroacetic acid and 15 ml of dichloromethane, and the mixture was stirred at 25° C. for 12 hours. After the reaction was completed, it was concentrated, and the residue was added with 20 ml of methyl tert-butyl ether and stirred at room temperature for 10 min, filtered, and the filter cake was dried to obtain the trifluoroacetate salt of compound M9. 1 H NMR (400MHz, D2O) δ = 4.30 (d, J = 3.8Hz, 1H), 3.97 (q, J = 4.9Hz, 1H), 3.73 (s, 3H), 2.93-2.83 (m, 1H), 2.54 (ddd, J = 4.5, 6.5, 11.5Hz, 1H), 2.39 (td, J = 5.7, 11.7Hz, 1H), 2.04 (dd, J = 9.4, 11.7Hz, 1H), 1.76 (dd, J = 9.5, 12.0Hz, 1H).

[0443] Reference Example 9: Compound M10

[0444]

[0445] Step 1

[0446] Compound M4-4 (9.2 g, 42.58 mmol), bis-naphthalene borate (16.22 g, 63.87 mmol), 1,5-cyclooctadiene iridium chloride dimer (858 mg, 1.28 mmol), 4,4'-di-tert-butyl-2,2'-bipyridine (1.71 g, 6.39 mmol) were dissolved in tetrahydrofuran (200 mL), replaced with nitrogen 3 times, and the reaction solution was stirred at 70 ° C for 16 hours. After the reaction was completed, the reaction solution was concentrated, then diluted with 150 ml of water and 150 ml of ethyl acetate, and then alkaline water (7.5 g of sodium hydroxide and 30 g of sodium carbonate in 400 ml of aqueous solution) was added to adjust the pH to 10, separated, and the organic phase was discarded. The aqueous phase was adjusted to pH 6 with concentrated hydrochloric acid, and then extracted with ethyl acetate (100 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound M10-1.

[0447] Step 2

[0448] Compound M10-1 (5 g, 19.24 mmol) was dissolved in acetonitrile (50 mL), and cuprous iodide (733 mg, 3.85 mmol), potassium iodide (6.39 g, 38.48 mmol), potassium carbonate (5.32 g, 38.48 mmol), and 1,10-phenanthroline (694 mg, 3.85 mmol) were added in sequence. The reaction solution was stirred at 60 ° C for 2 hours. After the reaction was completed, the reaction solution was filtered and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-10%) to obtain compound M10-2. LCMS: m / z = 341.8, 343.8 [M+1] + .

[0449] Step 3

[0450] Compound M10-2 (0.27 g, 789.54 μmol) and compound M10-3 (170 mg, 790.54 μmol, 2HCl) were dissolved in toluene (5 mL), cooled to 0°C, and then cesium carbonate (1.29 g, 3.95 mmol), (R)-(+)-2,2-bis(diphenylphosphino)-1,1-binaphthyl (50 mg, 80.95 μmol) and palladium acetate (36 mg, 158.91 μmol) were added. Nitrogen was replaced three times, and the reaction solution was stirred at 90°C for 12 hours. After the reaction was completed, the reaction solution was directly concentrated, and the crude product was purified by flash chromatography (silica gel, eluent methanol / dichloromethane, methanol ratio: 0-10%) to obtain compound M10. LCMS: m / z=356.0,358.0[M+1] + .

[0451] Reference Example 10: Compound M11

[0452]

[0453] Step 1

[0454] Compound M11-1 (5 g, 20.58 mmol) and potassium carbonate (8.53 g, 61.75 mmol) were dissolved in methylpyrrolidone (25 mL), and then tert-butyl cyanoacetate (4.65 g, 32.93 mmol) was added. The reaction was stirred at 85 °C for 12 hours. After the reaction was completed, it was cooled to room temperature and water (25 mL) was added. The pH was adjusted to 2 with 3M hydrochloric acid, and the solid was precipitated. The solid was filtered and the filter cake was dried to obtain compound M11-2. 1 H NMR (400MHz, CD3OD) δppm 1.53 (s, 9H), 4.85 (s, 1H), 6.93 (s, 1H).

[0455] Step 2

[0456] Compound M11-2 (6.2 g, 20.45 mmol) was dissolved in hydrochloric acid (4M, 24 mL), and then acetic acid (439.68 mmol, 25.17 mL) was added and stirred at 80 ° C for 15 minutes. After the reaction was completed, water (100 mL) was added and extracted with ethyl acetate (150 mL*3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:1 to 3:1) to obtain compound M11-3. 1 HNMR (400MHz, CD3OD) δppm 4.90 (s, 2H), 7.61 (s, 1H).

[0457] Step 3

[0458] Compound M11-3 (3 g, 14.77 mmol) was dissolved in tetrahydrofuran (30 mL), sodium hydrogen (1.48 g, 36.93 mmol, purity 60%) was added at 0°C, stirred at 0°C for 20 minutes under nitrogen protection, and then 1,2-dibromoethane (22.16 mmol, 1.67 mL) was added, and stirred at 25°C for 12 hours under nitrogen protection. After the reaction was completed, water (100 mL) was added to quench, and ethyl acetate (150 mL*3) was used for extraction. The organic phase was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (petroleum ether: ethyl acetate = 10:1 to 3:1) to obtain compound M11-4. 1 HNMR (400MHz, CD3OD) δppm 1.78-1.83 (m, 2H) 1.84-1.89 (m, 2H) 7.07 (s, 1H).

[0459] Step 4

[0460] Compound M11-4 (1 g, 4.36 mmol) was dissolved in tetrahydrofuran (10 mL), and then diisobutylaluminum hydride toluene solution (1 M, 10.91 mL) was added at 0°C and stirred at 0°C for 1 hour under nitrogen protection. After the reaction was completed, water (120 mL) was added to quench the reaction, and the mixture was extracted with ethyl acetate (50 mL*3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain a crude product. The crude product was purified by column chromatography (petroleum ether: ethyl acetate = 10:1 to 3:1) to obtain compound M11-5. 1 H NMR (400MHz, DMSO-d6) δppm 1.97-2.01(m,2H)2.07-2.11(m,2H)7.79(s,1H)8.90(s,1H).

[0461] Step 5

[0462] Compound M11-5 (0.2 g, 861.71 μmol) and ammonium carbonate (248.39 mg, 2.59 mmol) were dissolved in ethanol (5 mL) and water (5 mL), potassium cyanide (84.17 mg, 1.29 mmol) was added, and the mixture was stirred at 80°C for 6 hours. After the reaction was completed, water (150 mL) was added to quench the mixture, and the mixture was extracted with ethyl acetate (150 mL*3). The organic phase was dried over anhydrous sodium sulfate, and then concentrated under reduced pressure to obtain a crude product, thereby obtaining compound M11-6. LCMS: m / z=302.1,304.1[M+1] + . 1 H NMR (400MHz, CD3OD) δppm 1.20-1.32(m,2H)1.37-1.50(m,2H)4.06(s,1H)7.39(s,1H).

[0463] Step 6

[0464] Compound M11-6 (0.17 g, 562.64 μmol) was dissolved in dioxane (1 mL) and water (1 mL), and barium hydroxide (385.61 mg, 2.25 mmol) was added. The reaction system was subjected to microwave reaction at 130°C for 30 minutes. Water (50 mL) was added to the reaction system, and the mixture was washed with ethyl acetate (50 mL*3). The aqueous phase was collected to obtain an aqueous solution of compound M11. LCMS: m / z=277.1, 279.1 [M+1] + .

[0465] Reference Example 11: Compound M12

[0466]

[0467] Step 1

[0468] Compound M10-2 (0.1 g, 292.42 μmol), compound M12-1 (108 mg, 350.91 μmol), potassium carbonate (101 mg, 731.06 μmol), 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (21 mg, 29.24 μmol) were dissolved in dioxane (5 mL) and water (1 mL), and nitrogen was replaced three times. The reaction solution was stirred at 60 ° C for 3 hours. After the reaction was completed, the reaction solution was directly concentrated, and the crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-40%) to obtain compound M12-2. LCMS: m / z = 397.1, 399.0 [M+1] + .

[0469] Step 2

[0470] Trimethyl sulfoxide iodide (139 mg, 629.24 μmol) and potassium tert-butoxide (71 mg, 629.24 μmol) were added to dimethyl sulfoxide (3 mL) and stirred at 50 ° C for 2 hours. Then the temperature was lowered to 25 ° C and a solution of compound M12-2 (0.05 g, 125.85 μmol) in dimethyl sulfoxide (1 mL) was added dropwise, and the reaction solution was stirred at 80 ° C for 12 hours. After the reaction was completed, water (50 mL) was added to dilute the reaction, and the solution was extracted with ethyl acetate (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%) to obtain compound M12. LCMS: m / z=410.9,412.9[M+1] + .

[0471] Example 1

[0472]

[0473]

[0474] Step 1

[0475] To a mixed solution of compound M4 (5 g, 19.00 mmol) and compound M5 (7.02 g, 10.86 mmol) in 70 ml of dioxane and 15 ml of water, K2CO3 (3.75 g, 27.14 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (794.48 mg, 1.09 mmol) were added, the atmosphere was replaced with nitrogen three times, and the mixture was stirred at 85°C for 4 hours. The reaction solution was concentrated. The residue was diluted with 100 ml of water, extracted with ethyl acetate (3*50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to obtain compound 1-1. LCMS: m / z=655.1,657.1[M+1] + .

[0476] Step 2

[0477] At 0°C, ethyl iodide (2.14 g, 13.73 mmol) and cesium carbonate (4.47 g, 13.73 mmol) were added to a DMF (50 mL) solution of compound 1-1 (4.5 g, 6.86 mmol) in sequence. After the addition, the temperature was raised to 25°C and stirred at 25°C for 16 hours. The reaction solution was diluted with 100 ml of water and extracted with ethyl acetate (2*50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 1-2. LCMS: m / z=683.2,685.2[M+1] + .

[0478] Step 3

[0479] Compound 1-2 (3.4 g) and tetrabutylammonium fluoride tetrahydrofuran solution (1M, 34.81 mL) were added to 35 mL of anhydrous tetrahydrofuran, and the mixture was stirred at 50°C for 16 hours. The reaction solution was diluted with 100 mL of water, extracted with ethyl acetate (2*50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-80%) to obtain compound 1-3A (TLC developing solvent: ethyl acetate, R of compound 1-3A f =0.38, its isomer R f =0.17). LCMS: m / z=445.1,447.1[M+1] + ; 1 H NMR (400MHz, CDCl3) δ8.85 (dd, J=1.76, 4.77Hz, 1H), 7.92 (d, J=1.76Hz, 1H), 7.71 (dd ,J=1.76,7.78Hz,1H),7.33-7.41(m,2H),7.24-7.28(m,1H),4.08-4.14(m,1H),3.96- 4.08(m,1H),3.83-3.96(m,1H),3.19-3.33(m,2H),3.09(s,3H),2.74(d,J=14.05Hz, 1H), 2.27 (d, J = 14.05Hz, 1H), 1.50 (d, J = 6.27Hz, 3H), 1.16-1.24 (m, 4H), 0.80 (s, 6H).

[0480] Step 4

[0481] Compound 1-3A (0.4 g, 898.09 μmol) and neopentyl glycol diboronate (406 mg, 1.80 mmol) were dissolved in toluene (10 mL), and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (131 mg, 179.62 μmol) and potassium acetate (264 mg, 2.69 mmol) were added in sequence. The nitrogen was replaced three times, and the reaction solution was heated to 90°C and stirred for 12 hours. The reaction solution was filtered and the filtrate was concentrated to obtain compound 1-4A. LCMS: m / z=411.2 [M Boric acid + 1] + .

[0482] Step 5

[0483] Compound 1-4A (300.00 mg, 627.05 μmol), compound M2 (0.36 g, 715.12 μmol), 1,1-di(tert-butylphosphino)ferrocenepalladium chloride (47 mg, 71.51 μmol), potassium phosphate (455 mg, 2.15 mmol) were dissolved in toluene (6 mL), dioxane (2 mL) and water (2 mL), and nitrogen was replaced three times. The reaction solution was stirred at 70 ° C for 12 hours. The reaction solution was concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to obtain compound 1-5A. LCMS: m / z = 789.4 [M+1] + .

[0484] Step 6

[0485] Compound 1-5A (100.00 mg, 126.74 μmol) was dissolved in tetrahydrofuran (2 mL) and water (0.2 mL), and then lithium hydroxide monohydrate (16 mg, 380.23 μmol) was added. The reaction solution was stirred at 15°C for 12 hours. 1M HCl was added to the reaction solution to adjust the pH to 5, and the solution was extracted with ethyl acetate (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 1-7A. LCMS: m / z=761.3[M+1] + .

[0486] Step 7

[0487] Compound 1-7A (50.00 mg, 65.71 μmol) was dissolved in dichloromethane (5 mL), and then N, N-diisopropylethylamine (260 mg, 2.01 mmol, 0.35 mL), 1-hydroxybenzotriazole (0.044 g, 325.63 μmol), and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.35 g, 1.83 mmol) were added. The reaction solution was stirred at 20 ° C for 12 hours. Water (50 mL) was added to dilute the reaction solution, and the solution was extracted with dichloromethane (20 mL*3). The organic phases were combined and washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-80%) to obtain compound 1-8A. LCMS: m / z = 743.4 [M+1] + .

[0488] Step 8

[0489] At 0°C, compound 1-8A (20.00 mg, 26.92 μmol) was dissolved in dichloromethane (1 mL), and then trifluoroacetic acid (1.54 g, 13.46 mmol, 1 mL) was added. The reaction solution was stirred at 15°C for 12 hours. The reaction solution was concentrated to obtain a crude trifluoroacetate salt of compound 1-9A. LCMS: m / z = 643.3 [M+1] + .

[0490] Step 9

[0491] Compound 1-9A (20.00 mg, crude trifluoroacetate) was dissolved in N,N-dimethylformamide (2 mL), and then N,N-diisopropylethylamine (311.13 μmol, 54 μL), (1S,2S)-2-methylcyclopropane-1-carboxylic acid (7 mg, 62.3 μmol) and O-(7-azabenzotriazole-1-YL)-N,N,N,N-tetramethyluronium hexafluorophosphonate (36 mg, 93.4 μmol) were added. The reaction solution was stirred at 20°C for 1.5 hours. Water (20 mL) was added to dilute the reaction solution, and the solution was extracted with ethyl acetate (20 mL*3). The organic phases were combined and washed once with saturated brine (20 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product, which was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-80%) to obtain compound 1A. LCMS: m / z = 725.3 [M+1] + ; 1H NMR(400MHz,CD3OD)δppm 8.64(dd,J=4.8,1.6Hz,1H),8.12(s,1H),7.79(dd,J=7.6,1.6Hz,1H),7.54(d,J=8.4Hz,1H),7.44(dd,J=7.6,4.8Hz ,1H),7.34-7.39(m,2H),6.02(t,J=6.2Hz,1H),4.03-4.13(m,2H),3.71-3.86(m,3H),3.43-3.56(m,2H),3.03(s,3H ),2.64-2.72(m,1H),2.37-2.44(m,1H),2.06-2.13(m,1H),1.89-1.97(m,2H),1.70-1.80(m,1H),1.48-1.60(m,3H) ,1.32-1.37(m,2H),1.18-1.23(m,6H),0.90-0.97(m,2H),0.76-0.83(m,2H),0.57-0.66(m,6H),0.45-0.51(m,1H).

[0492] Example 2

[0493]

[0494] Step 1

[0495] Compound 1-3A (295 mg, 662.34 μmol) and bis-naphthalene borate (252.29 mg, 993.51 μmol) were dissolved in toluene (2 mL), potassium acetate (130.01 mg, 1.32 mmol) and [1,1-bis(diphenylphosphino)ferrocene] palladium dichloride (48.46 mg, 66.23 μmol) were added, and the reaction solution was stirred at 110°C under nitrogen atmosphere for 12 hours. The reaction solution was filtered and the filtrate was concentrated under reduced pressure to obtain compound 2-1A. LCMS: m / z=493.3[M+1] + .

[0496] Step 2

[0497] Compound 2-1A (300 mg, 609.19 μmol) and compound M3 (492.89 mg, 913.79 μmol) were dissolved in a mixed solvent of 1,4-dioxane (3 mL), toluene (3 mL), and water (0.5 mL), potassium phosphate (387.94 mg, 1.83 mmol) and 1,1-di(tert-butylphosphino)ferrocenepalladium chloride (39.70 mg, 60.92 μmol) were added, and the reaction solution was stirred at 70°C in a nitrogen atmosphere for 12 hours. The reaction solution was filtered, and the filtrate was concentrated to obtain a crude product, which was purified by silica gel column chromatography (20-80% ethyl acetate / petroleum ether) to obtain compound 2-2A. LCMS: m / z=825.1[M+1] + .

[0498] Step 3

[0499] Compound 2-2A (280 mg, 339.40 μmol) was dissolved in a mixed solution of tetrahydrofuran (3 mL) and water (1 mL), and lithium hydroxide monohydrate (71.21 mg, 1.70 mmol) was added. The reaction solution was stirred at 25°C for 2 hours, and then the pH of the reaction solution was adjusted to neutral with 1M dilute hydrochloric acid, and then extracted with ethyl acetate (5 mL×3). The organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain compound 2-3A. LCMS: m / z=797.2[M+1] + .

[0500] Step 4

[0501] Compound 2-3A (0.3 g, 376.45 μmol) was dissolved in dichloromethane (30 mL), 1-hydroxybenzotriazole (508.67 mg, 3.76 mmol), N,N-diisopropylethylamine (11.29 mmol, 1.97 mL), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (2.16 g, 11.29 mmol) were added, and the reaction solution was stirred at 25°C for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography (0-30% ethyl acetate / petroleum ether), and then separated by high performance liquid chromatography (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient (acetonitrile%): 39%-69%) to obtain compound 2-4A. LCMS: m / z=779.3[M+1] + .

[0502] Step 5

[0503] Compound 2-4A (20 mg, 25.68 μmol) was dissolved in dichloromethane (1 mL), trifluoroacetic acid (25.68 μmol, 1.91 μL) was added, the reaction solution was stirred at 25°C for 5 hours, and concentrated under reduced pressure to obtain the crude trifluoroacetate salt of compound 2-5A. LCMS: m / z=679.3[M+1] + .

[0504] Step 6

[0505] Compound 2-5A (17 mg, trifluoroacetate) and (1S,2S)-2-methylcyclopropane-1-carboxylic acid (3.76 mg, 37.57 μmol) were dissolved in N,N-dimethylformamide (1 mL), and then N,N-diisopropylethylamine (9.71 mg, 75.13 μmol, 13.09 μL) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexahydrate were added under stirring. Fluorophosphonate (19.05 mg, 50.09 μmol), the reaction solution was stirred at 25°C for 12 hours, extracted with ethyl acetate (5 mL×3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was prepared by HPLC (column: C18100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient (acetonitrile%): 27%-57%) to obtain the trifluoroacetate salt of compound 2A. LCMS: m / z=761.2[M+1] + ; 1 H NMR(CDCl3,400MHz)δ9.1-9.2(m,1H),8.3-8.4(m,1H),8.0-8.2(m,1H),7.7-7.8(m,1H),7 .6-7.7(m,1H),7.3-7.5(m,1H),6.6-6.7(m,1H),5.7-5.8(m,1H),4.4-4.5(m,1H),4.17(br d,J=5.8Hz,1H),4.11(br d,J=10.5Hz,1H),4.0-4.1(m,1H),3.88(br d,J=10.3Hz,1H),3.64(br d,J=15.6Hz,1H),3.1-3.3(m,6H),2.1-2.2(m,1H),1.9-2.0(m,2H),1.6-1.7(m,1H),1.52(br d,3H,J=5.8Hz),1.2-1.4(m,8H),1.1-1.2(m,1H),1.07(br d,3H,J=5.5Hz),0.92(br s,3H),0.6-0.7(m,3H),0.5-0.6(m,1H).

[0506] Example 3

[0507]

[0508]

[0509] Step 1

[0510] Compound M4-4 (30 g, 138.84 mmol), bis-naphthalene borate (52.89 g, 208.26 mmol), 1,5-cyclooctadiene iridium chloride dimer (2.80 g, 4.17 mmol) and 4,4'-di-tert-butyl-2,2'-bipyridine (5.59 g, 20.83 mmol) were added to 600 ml of anhydrous tetrahydrofuran, replaced with nitrogen three times, and the mixture was stirred at 80°C for 20 hours. The reaction solution was concentrated. The residue was diluted with 300 ml of water and 200 ml of ethyl acetate, and alkaline water (10 g of NaOH and 40 g of sodium carbonate in 400 ml of aqueous solution) was added to adjust the pH to 10, separated, and the organic phase was discarded. The aqueous phase was adjusted to pH 6 with concentrated hydrochloric acid, and 500 ml of ethyl acetate was added for extraction three times. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 3-1. 1 H NMR (400MHz, CDCl3) δ=8.93 (d, J=1.6Hz, 1H), 8.22 (d, J=1.6Hz, 1H), 4.96 (q, J=6.4Hz, 1H), 3.32 (s, 3H), 1.50 (d, J=6.4Hz, 3H), 1.37 (s, 12H).

[0511] Step 2

[0512] At 80°C, compound 3-1 (40 g, 116.95 mmol) was slowly added dropwise to a 1.5 liter acetonitrile suspension of benzyl-1-piperazine carbonate (77.28 g, 350.84 mmol), copper acetate (21.24 g, 116.95 mmol) and triethylamine (116.95 mmol, 16.28 mL). The mixture was stirred at 80°C for 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-30%) to obtain compound 3-2. MS-ESI calculated value [M+1] + 434.1, 436.1 Measured values ​​434.3, 436.3; 1H NMR (400MHz, CDCl3) δ = 8.23 ​​(d, J = 2.4Hz, 1H), 7.39-7.25 (m, 5H), 7.23 (d, J = 2.4Hz ,1H),5.09(s,2H),4.78(q,J=6.4Hz,1H),3.68-3.55(m,4H),3.21(s,3H),3.13(br s, 4H), 1.39 (d, J = 6.4Hz, 3H).

[0513] Step 3

[0514] Compound 3-2 (7 g, 16.12 mmol), neopentyl glycol diboron (5.46 g, 24.18 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (589.65 mg, 805.85 μmol) and potassium acetate (3.95 g, 40.29 mmol) were added to 140 ml of anhydrous dioxane, replaced with nitrogen three times, and the mixture was stirred at 80°C for 20 hours. The reaction solution was concentrated. The residue was purified by flash chromatography (silica gel, eluent methanol / dichloromethane, methanol ratio: 0-10%) to obtain compound 3-3. LCMS: m / z=400.1[M+1] + .

[0515] Step 4

[0516] Compound 3-3 (9 g, 22.54 mmol), compound M5 (14.57 g, 22.54 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (824.72 mg, 1.13 mmol) and potassium phosphate (11.96 g, 56.36 mmol) were added to 250 ml of anhydrous dioxane and 80 ml of water in sequence, replaced with nitrogen three times, and the mixture was stirred at 70°C for 12 hours. The reaction solution was concentrated. The residue was diluted with 200 ml of water, extracted with ethyl acetate (3*200 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-80%) to obtain compound 3-4. LCMS: m / z=873.3,875.3[M+1] + .

[0517] Step 5

[0518] Compound 3-4 (6.2 g, 7.09 mmol), iodoethane (14.19 mmol, 1.13 mL) and cesium carbonate (4.62 g, 14.19 mmol) were added to 100 mL of anhydrous DMF in sequence, and the mixture was stirred at 20°C for 10 hours. The reaction solution was concentrated. The residue was diluted with 100 mL of water, extracted with ethyl acetate (3*150 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 3-5. LCMS: m / z=901.4,903.4[M+1] + .

[0519] Step 6

[0520] Compound 3-5 (6.2 g, 6.87 mmol) and tetrabutylammonium fluoride tetrahydrofuran solution (1M, 13.75 mL) were added to 100 mL of anhydrous tetrahydrofuran, and the mixture was stirred at 50°C for 15 hours. The reaction solution was concentrated. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to obtain compound 3-6A (TLC developing solvent: ethyl acetate, R of compound 3-6A) f =0.43, its isomer R f =0.33). LCMS: m / z=663.2,665.2[M+1] + .

[0521] Step 7

[0522] Compound 3-6A (1.70 g, 2.56 mmol), bis-naphthalene borate (975.74 mg, 3.84 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (187.43 mg, 256.16 μmol) and potassium acetate (754.21 mg, 7.68 mmol) were added to 50 ml of anhydrous dioxane, replaced with nitrogen three times, and the mixture was stirred at 90 ° C for 20 hours. The reaction solution was concentrated. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to obtain compound 3-7A. LCMS: m / z=711.1[M+1] + .

[0523] Step 8

[0524] Compound 3-7A (0.38 g, 534.68 μmol), compound M2 (322.99 mg, 641.61 μmol), 1,1-di(tert-butylphosphino)ferrocenepalladium chloride (34.85 mg, 53.47 μmol) and potassium phosphate (283.74 mg, 1.34 mmol) were added to 15 ml of dioxane and 5 ml of water in sequence, replaced with nitrogen three times, and the mixture was stirred at 70°C for 4 hours. The reaction solution was concentrated. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to obtain compound 3-8A. LCMS: m / z=1007.4[M+1] + .

[0525] Step 9

[0526] Compound 3-8A (650 mg, 645.33 μmol) and lithium hydroxide monohydrate (54.16 mg, 1.29 mmol) were added to 20 ml of tetrahydrofuran and 20 ml of water in sequence, and the mixture was stirred at 15°C for 20 hours. The reaction solution was concentrated. The residue was diluted with 5 ml of water, 1N dilute hydrochloric acid was added dropwise to adjust the pH to 6-7, and extracted with ethyl acetate (3*30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 3-9A. LCMS: m / z=979.4[M+1] + .

[0527] Step 10

[0528] Compound 3-9A (650 mg, 663.81 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (3.82 g, 19.91 mmol), N,N-diisopropylethylamine (26.55 mmol, 4.62 mL) and 1-hydroxybenzotriazole (896.94 mg, 6.64 mmol) were added to 65 ml of acetonitrile in sequence, and the mixture was stirred at 25°C for 40 hours. The reaction solution was concentrated. The residue was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to obtain compound 3-10A. LCMS: m / z=961.4[M+1] + .

[0529] Step 11

[0530] Compound 3-10A (150 mg, 156.06 μmol), paraformaldehyde (23.45 mg, 780.30 μmol) and palladium hydroxide on carbon (100 mg, 10% w / w, 50% water content) were added to 10 ml of methanol in sequence, and hydrogen was replaced 3 times. The mixture was stirred at 20°C for 2 hours under a hydrogen atmosphere (15 psi). Filtered. The filtrate was concentrated. The residue was purified by flash chromatography (silica gel, eluent methanol / dichloromethane, methanol ratio: 0-10%) to obtain compound 3-11A. LCMS: m / z=841.4[M+1] + .

[0531] Step 12

[0532] 1 ml of trifluoroacetic acid was slowly added dropwise to 5 ml of anhydrous dichloromethane of compound 3-11A (56 mg, 58.26 μmol) at 0°C, and the mixture was stirred at 0°C for 5 hours. The reaction solution was concentrated to obtain the trifluoroacetate salt of compound 3-12A. LCMS: m / z=741.3[M+1] + .

[0533] Step 13

[0534] Compound 3-12A (45 mg, trifluoroacetate), compound (1S,2S)-2-methylcyclopropane-1-carboxylic acid (9.12 mg, 91.10 μmol), N,N-diisopropylethylamine (303.66 μmol, 52.89 μL) and HATU (69.28 mg, 182.20 μmol) were added to 5 ml of anhydrous DMF in sequence, and the mixture was stirred at 20°C for 1 hour. The reaction solution was concentrated, and the crude product was separated by preparative high performance liquid chromatography (preparation method: column model: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient (acetonitrile%): from 14% to 44% in 8 minutes) to obtain the trifluoroacetate of compound 3A. LCMS: m / z=823.4[M+1] + ; 1H NMR (400MHz, CDCl3) δ=8.67(br s,1H),8.38(s,1H),7.57(br d,J=8.4Hz,1H),7.37(d,J=8.4Hz,1H),7.27(s,1H),7.20(s,1H),6.78(br d,J=7.6Hz,1H),5.91(br s,1H),4.22(br d,J=6.0Hz,1H),4.14-4.06(m,2H),3.99-3.91(m,2H),3.67(brs,4H),3.53(br dd,J=5.6,14.8Hz,4H),3.27(s,3H),3.26-2.99(m,3H),2.92(br s,3H),2.81(br d,J=14.4Hz,1H),2.52(br d,J=14.2Hz,1H),2.12-1.71(m,4H),1.47(br d,J=6.0Hz,3H),1.43-1.14(m,6H),1.14-1.06(m,6H),0.79(s,3H),0.66(s,3H),0.63(br s,1H).

[0535] Example 4

[0536]

[0537] Compound 3-12A (0.1 g, 134.96 μmol) and (1r, 2R, 3S)-2,3-dimethylcyclopropyl-1-carboxylic acid (31 mg, 269.98 μmol) were dissolved in N,N-dimethylformamide (2 mL), and then N,N-diisopropylethylamine (175 mg, 1.35 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (154 mg, 404.9 μmol) were added. The reaction solution was stirred at 20°C for 2 hours. The reaction solution was filtered, and the filtrate was directly purified by HPLC preparation (purification method: column model: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient (acetonitrile%): 16% to 46% in 8 minutes) to obtain the trifluoroacetate salt of compound 4A. LCMS: m / z = 837.4 [M+1] + ; 1H NMR (400MHz, CD3OD) δppm8.53(s,1H),8.27(s,1H),7.93(s,1H),7.70(d,J=8.8Hz,1H),7.49-7.57(m,2H),6.1 6(t,J=6.0Hz,1H),4.95-5.04(m,3H),4.10-4.28(m,3H),3.88-4.06(m,4H),3.45-3.70(m,3H),3.24(s,3H),3. 02(s,3H),2.82-2.90(m,1H),2.46-2.55(m,1H),2.01-2.11(m,1H),1.81-1.91(m,1H),1.68-1.74(m,1H),1.5 8-1.66(m,1H),1.42-14.7(m,3H),1.26-1.41(m,7H),1.17-1.23(m,3H),1.08-1.15(m,7H),0.75-0.85(m,6H).

[0538] Example 5

[0539]

[0540] Step 1

[0541] Compound M8 (200 mg, 338.64 μmol) and compound M3 (274 mg, 507.98 μmol) were dissolved in dioxane (4 mL), toluene (1.3 mL) and water (1.3 mL), potassium phosphate (215.65 mg, 1.02 mmol) and 1,1-di(tert-butylphosphino)ferrocenepalladium chloride (22.07 mg, 33.86 μmol) were added, and the reaction solution was stirred at 70°C under a nitrogen atmosphere for 12 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to obtain a crude product, which was purified by column chromatography (0-15% methanol / dichloromethane) to obtain compound 5-1A. LCMS: m / z=923.5[M+1] + .

[0542] Step 2

[0543] Compound 5-1A (205 mg, 222.07 μmol) was dissolved in a mixed solution of tetrahydrofuran (2 mL) and water (0.5 mL), and lithium hydroxide monohydrate (46.59 mg, 1.11 mmol) was added. The reaction solution was stirred at 25°C for 1 hour, and then the reaction solution was adjusted to pH 7-8 with 1M dilute hydrochloric acid, and then extracted with ethyl acetate (10 mL*5). The organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound 5-2A, which was directly used in the next step. LCMS: m / z=895.3[M+1]+ .

[0544] Step 3

[0545] Compound 5-2A (45 mg, 50.28 μmol) was dissolved in acetonitrile (4.5 mL), tetramethyl chlorourea hexafluorophosphate (21.16 mg, 75.41 μmol) and N-methylimidazole (12.38 mg, 150.83 μmol) were added, and the reaction solution was stirred at 25°C for 1 hour. Water (20 mL) was added to the reaction solution, and it was extracted with dichloromethane (5 mL*3). The organic phase was washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated to obtain compound 5-3A. LCMS: m / z=877.4[M+1] + .

[0546] Step 4

[0547] Compound 5-3A (50 mg, 57.01 μmol) was dissolved in dichloromethane (0.5 mL), trifluoroacetic acid (767.50 mg, 6.73 mmol, 0.5 mL) was added, the reaction solution was stirred at 25 °C for 1 hour, and concentrated under reduced pressure to obtain the crude trifluoroacetate salt of compound 5-4A, which was directly used in the next step.

[0548] Step 5

[0549] The crude trifluoroacetate of compound 5-4A obtained in step 4 was dissolved in N,N-dimethylformamide (1 mL), (1r,2R,3S)-2,3-dimethylcyclopropyl-1-carboxylic acid (9.70 mg, 84.95 μmol) was added, and then N,N-diisopropylethylamine (21.96 mg, 169.90 μmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphonate (43.07 mg, 113.27 μmol) were added under stirring. The reaction solution was stirred at 25° C. for 3 hours, and then purified by prep-HPLC (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: the proportion of acetonitrile increased from 16% to 46% within 8 minutes) to obtain the trifluoroacetate of compound 5A. LCMS: m / z = 873.9 [M+1] +. 1H NMR(CD3OD,400MHz)δ8.5-8.6(m,1H),8.2-8.3(m,1H),7.7-7.7(m,1H),7.61(br d,J=2.3Hz,1H),7.4-7.5(m,2H),5.5-5.6(m,3H),5.3-5.4(m,3H),4.0-4.3(m,6H),3.8-3.9(m,2H),3.4-3.6(m,5H),3.0-3.1 (m,5H),2.2-2.3(m,3H),2.1-2.2(m,2H),2.0-2.1(m,6H),1.6-1.7(m,3H),1.4-1.5(m,3H),1.0-1.1(m,3H),0.8-1.0(m,6H).

[0550] Example 6

[0551]

[0552]

[0553] Step 1

[0554] Compound M8 (120 mg, 203.18 μmol) and M1-1 (89.05 mg, 243.82 μmol) were dissolved in toluene (9 mL), 1,4-dioxane (3 mL) and water (3 mL). Potassium phosphate (129.39 mg, 609.55 μmol) and [1,1'-bis(di-tert-butylphosphino)ferrocene]palladium dichloride (26.48 mg, 40.64 μmol) were added under nitrogen protection, and stirred at 70°C for 12 hours. After the reaction solution was cooled, it was directly concentrated and purified by column chromatography (eluent: ethyl acetate / petroleum ether 3% to 7%) to obtain compound 6-1A. LCMS: m / z=749.6[M+1] + .

[0555] Step 2

[0556] Compound 6-1A (130 mg, 173.57 μmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (3 mL), and lithium hydroxide monohydrate (36.42 mg, 867.86 μmol) was added, and stirred at 25°C for half an hour. The pH value of the system was adjusted to 7.0 with 1M hydrochloric acid, and then diluted with saturated brine (50 mL), extracted twice with ethyl acetate (30 mL) and tetrahydrofuran (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 6-2A, which was directly used in the next step. LCMS: m / z=735.6[M+1] + .

[0557] Step 3

[0558] Compound M6 (21.82 mg, 95.25 μmol) and compound 6-2A (70 mg, 95.25 μmol) were dissolved in N, N-dimethylformamide (2 mL), and N, N-diisopropylethylamine (952.45 μmol, 165.90 μL) and 2-(7-azobenzotriazole)-N, N, N', N'-tetramethyluronium hexafluorophosphate (43.46 mg, 114.29 μmol) were added, and stirred at 25 ° C for half an hour. After the reaction was completed, saturated brine (30 mL) was added, and extracted with ethyl acetate (30 mL) and tetrahydrofuran (30 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 6-3A, which was directly used in the next step. LCMS: m / z=874.0[M+1] + .

[0559] Step 4

[0560] Compound 6-3A (130 mg, 173.57 μmol) was dissolved in tetrahydrofuran (3 mL), methanol (1 mL) and water (3 mL), and lithium hydroxide monohydrate (36.42 mg, 867.86 μmol) was added, and stirred at 25°C for half an hour. The pH value of the system was adjusted to 7.0 with 1M hydrochloric acid, and then diluted with saturated brine (50 mL), extracted twice with ethyl acetate (30 mL) and tetrahydrofuran (30 mL), and the organic phases were combined and dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 6-4A, which was directly used in the next step. LCMS: m / z=860.0[M+1] + .

[0561] Step 5

[0562] Compound 6-4A (61 mg, 71.01 μmol) was dissolved in acetonitrile (20 mL), and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (597.68 mg, 2.13 mmol) and N-methylimidazole (174.89 mg, 2.13 mmol, 169.80 μL) were added, and stirred at 25 ° C for 1 hour under nitrogen protection. The reaction solution was concentrated and separated by pre-TLC preparation (DCM / MeOH=10:1) to obtain compound 6-5A. LCMS: m / z=842.0[M+1] + .

[0563] Step 6

[0564] Compound 6-5A (52 mg, 61.83 μmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (798.20 mg, 7.00 mmol, 520.00 μL) was added, and stirred at 25° C. for 1 hour. The reaction solution was concentrated to obtain the crude trifluoroacetate salt of compound 6-6A, which was used directly in the next step.

[0565] Step 7

[0566] Compound 6-6A (35 mg, 40.9 μmol) and (1r,2R,3S)-2,3-dimethylcyclopropyl-1-carboxylic acid (9.35 mg, 81.8 μmol)) were dissolved in DMF (2 mL), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (31.13 mg, 81.87 μmol) and N,N-diisopropylethylamine (409.37 μmol, 71.30 μL) were added, and the mixture was stirred at 25 °C for 1 hour. The mixture was diluted with saturated brine (50 mL), extracted twice with ethyl acetate (30 mL) and tetrahydrofuran (30 mL), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product which was purified by pre-TLC (developing solvent: dichloromethane / methanol = 10:1), and then separated and purified by SFC (column: DAICEL CHIRALCEL OD (250 mm*30 mm, 10 μm); mobile phase: [phase A is supercritical carbon dioxide, phase B is ethanol (0.1% ammonia water)]; gradient (B%): 50%, isobaric elution) to obtain compounds 6A and 6B.

[0567] According to SFC analysis (column: Chiralcel OD-3 50*4.6mm ID, 3um, mobile phase: A: supercritical carbon dioxide B: ethanol (containing 0.05% diethylamine), gradient elution: mobile phase B increased from 5% to 40% in two minutes, then maintained at 40% for 1.2 minutes, and then maintained at 5% for 0.8 minutes), the RT of compound 6A was 1.896min, ee=96.98%; the RT of compound 6B was 2.201min, ee=98.82%. Compound 6A: LCMS: m / z=837.5[M+1] +; 1H NMR (400 MHz, CD3OD) δ = 8.33 (br d, J = 10.5 Hz, 2H), 7.59 (br d, J = 8.3 Hz, 1H), 7.45 (s, 1H), 7.39 (br d, J = 8.5 Hz, 1H), 7.25 (br s, 1H), 5.48 (br s, 1H), 4.61 - 4.50 (m, 2H), 4.24 - 4.05 (m, 2H), 3.64 - 3.50 (m, 2H), 3.28 (br s, 3H), 3.21 (br s, 6H), 2.97 (br d, J = 13.3 Hz, 1H), 2.63 (br s, 4H), 2.58 - 2.44 (m, 2H), 2.33 (br s, 4H), 2.10 (br t, J = 9.0 Hz, 1H), 1.46 (br s, 1H), 1.38 - 1.16 (m, 8H), 1.05 (br dd, J = 6.0, 13.3 Hz, 6H), 0.93 - 0.85 (m, 3H), 0.81 (br s, 3H), 0.38 (br s, 3H).

[0568] Compound 6B: LCMS: m / z = 837.5 [M+1] + ; 1 1H NMR (400 MHz, CD3OD) δ = 8.57 (d, J = 2.8 Hz, 1H), 8.53 (s, 1H), 8.28 (d, J = 3.0 Hz, 1H), 7.76 (d, J = 9.8 Hz, 1H), 7.60 - 7.52 (m, 2H), 6.05 (dd, J = 3.6, 7.9 Hz, 1H), 4.64 (br d, J = 5.0 Hz, 1H), 4.49 (s, 1H), 4.32 - 4.08 (m, 5H), 3.97 - 3.86 (m, 2H), 3.79 - 3.61 (m, 3H), 3.41 - 3.35 (m, 4H), 3.24 (s, 3H), 3.01 (s, 3H), 2.70 (br d, J = 5.8 Hz, 1H), 2.57 - 2.49 (m, 1H), 2.48 - 2.41 (m, 1H), 2.38 - 2.28 (m, 1H), 2.17 (br d, J = 13.8 Hz, 1H), 1.80 (t, J = 9.3 Hz, 1H), 1.43 (d, J = 6.3 Hz, 3H), 1.37 - 1.25 (m, 6H), 1.09 (dd, J = 5.6, 9.7 Hz, 8H), 0.90 (s, 3H), 0.80 (s, 3H).

[0569] Example 7

[0570]

[0571]

[0572] Compound 6-6A (60 mg, 70.18 μmol) and (1S,2S)-2-methylcyclopropyl-1-carboxylic acid (14.05 mg, 81.8 μmol)) were dissolved in DMF (10 mL), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (53.37 mg, 140.35 μmol) and N,N-diisopropylethylamine (72.56 mg, 561.42 μmol) were added, and the mixture was stirred at 25 °C for 1 hour. The mixture was diluted with saturated brine (50 mL), extracted twice with ethyl acetate (30 mL) and tetrahydrofuran (30 mL), the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain a crude product which was purified by a fast silica gel column (mobile phase: dichloromethane / methanol = 100:1), and then separated and purified by SFC (column: DAICEL CHIRALCEL OD (250 mm*30 mm, 10 μm); mobile phase: [phase A is supercritical carbon dioxide, phase B is ethanol (0.1% ammonia water)]; gradient (B%): 50%) to obtain compounds 7A and 7B. According to SFC analysis (method: column: chiral OD-3 100*4.6mm ID, 3μm, mobile phase: A: supercritical carbon dioxide, B: ethanol (containing 0.05% diethylamine), gradient (B%): 40%), the RT of compound 7A is 1.365min, ee=100%; the RT of compound 7B is 2.325min, ee=88.2%. Compound 7A: LCMS: m / z=823.7[M+1] + ; 11H NMR (400 MHz, CD3OD) δ = 8.34 (s, 1H), 8.31 (d, J = 2.8 Hz, 1H), 7.58 (dd, J = 1.5, 8.8 Hz, 1H), 7.44 (s, 1H), 7.38 (d, J = 8.5 Hz, 1H), 7.24 (d, J = 3.0 Hz, 1H), 5.47 (br s, 1H), 4.60 - 4.44 (m, 3H), 4.22 - 4.03 (m, 3H), 3.64 - 3.50 (m, 2H), 3.31 - 3.26 (m, 4H), 3.25 (s, 2H), 3.02 - 2.91 (m, 1H), 2.61 (br t, J = 4.9 Hz, 5H), 2.53 - 2.46 (m, 1H), 2.38 - 2.32 (m, 1H), 2.31 (s, 3H), 2.09 (t, J = 9.9 Hz, 1H), 1.50 - 1.36 (m, 2H), 1.32 (d, J = 6.3 Hz, 3H), 1.21 - 1.11 (m, 3H), 1.08 (t, J = 7.0 Hz, 1H), 1.04 - 0.98 (m, 4H), 0.88 (br t, J = 7.0 Hz, 3H), 0.81 (s, 3H), 0.60 - 0.52 (m, 1H), 0.37 (s, 3H).

[0573] Compound 7B: LCMS: m / z = 823.7 [M+1] + ; 1H NMR (400MHz, CD3OD) δ = 8.38 (s, 1H), 8.33 (d, J = 2.8Hz, 1H), 7.60-7.54 (m, 1H), 7.39-7.34 (m, 2H), 7.31 (d, J = 3.0Hz, 1H), 5.98-5.88 (m, 1H) ,4.57-4.50(m,1H),4.37(s,1H),4.07-3.98(m,1H),3.96-3.88(m,2H),3.86-3.74(m,2H),3.55-3.46(m,1H),3.36-3.26(m,4H),2.97(br d,J=14.3Hz,1H),2.90(s,3H),2.62(br s,5H),2.46-2.38(m,1H),2.32(s,3H),2.26-2.11(m,2H),1.70(t,J=9.5Hz,1H),1.36-1.28(m,4H),1.20-1.11(m,1H),1.23-1.10 (m,5H),0.98-0.98(m,1H),0.99(d,J=6.0Hz,2H),0.91(td,J=4.3,8.5Hz,1H),0.69(s,3H),0.63-0.57(m,3H),0.50-0.41(m,1H).

[0574] Example 8

[0575]

[0576]

[0577] Using compound 8-1 and compound 6-6A as raw materials, refer to the synthesis method of Example 7, and the resulting reaction solution is directly purified by high performance liquid chromatography (column: C18 100×40mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile increased from 16% to 46% in 8 minutes) to obtain the trifluoroacetate salt of compound 8A. LCMS: m / z=859.5[M+1] + . 1H NMR(400MHz,DMSO-d6)δ=8.88(br d,J=9.0Hz,1H),8.51-8.47(m,1H),8.41(s,1H),7.84-7.80(m,1H),7.76-7.71(m,1H),7.59-7.55(m,1H),7.44-7.39(m,1H),6.01(br d,J=11.3Hz,1H),5.46-5.37(m,1H),5.34-5.30(m,1H),4.68-4.61(m,1H),4.53-4.47 (m,1H),4.37-4.23(m,1H),4.20-4.09(m,2H),4.06-3.99(m,2H),3.20(s,3H),3.05(br s,2H),2.95-2.89(m,1H),2.86(s,3H),2.65-2.58(m,1H),2.33(br d,J=1.5Hz,1H),2.20-2.13(m,1H),2.11-2.05(m,1H),2.04-1.94(m,3H),1.70-1.63(m,1H),1.62-1.54(m,1H),1.48-1.43(m,1H),1.34(br d,J=6.3Hz,4H),1.05(br t,J=6.9Hz,3H),0.94-0.83(m,10H),0.39-0.28(m,3H).

[0578] Example 9

[0579]

[0580]

[0581] Step 1

[0582] Compound M5-7 (6.83 g, 13.12 mmol) was dissolved in tetrahydrofuran (50 mL), and then tetrabutylammonium fluoride (1 M tetrahydrofuran solution, 65.60 mL) was added. The reaction solution was stirred at 50 ° C for 16 hours. After the reaction was completed, the reaction solution was directly concentrated. The crude product was diluted with water (100 mL), and the solution was extracted with ethyl acetate (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried with anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-55%) to obtain compound 9-1A. LCMS: m / z=281.9[M+1] + .

[0583] Step 2

[0584] Compound 9-1A (3.31 g, 11.73 mmol) was dissolved in dichloromethane (30 mL), and then triethylamine (3.56 g, 35.19 mmol) and 4-dimethylaminopyridine (72 mg, 596.51 μmol) were added. The temperature was lowered to 0°C, and acetic anhydride (1.16 g, 11.38 mmol) was added dropwise. The reaction solution was stirred at 0°C for 10 minutes. After the reaction was completed, water (50 mL) was added to dilute the reaction solution, and the solution was extracted with dichloromethane (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried with anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-30%) to obtain compound 9-2A. LCMS: m / z=323.9[M+1] + .

[0585] Step 3

[0586] Compound 9-2A (3.4 g, 10.49 mmol), bis-naphthalene borate (6.66 g, 26.22 mmol), potassium acetate (2.57 g, 26.22 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (0.768 g, 1.05 mmol) were dissolved in toluene 40 mL), replaced with nitrogen 3 times, and the reaction solution was stirred at 90 ° C for 3 hours. After the reaction was completed, the reaction solution was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-30%) to obtain compound 9-3A. LCMS: m / z = 372.1 [M+1] + .

[0587] Step 4

[0588] Compound 9-3A (3.8 g, 10.23 mmol), M1-1 (5.61 g, 15.35 mmol), potassium phosphate (5.43 g, 25.59 mmol), 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (750 mg, 1.12 mmol) were dissolved in toluene (30 mL), dioxane (10 mL) and water (10 mL), replaced with nitrogen 3 times, and the reaction solution was stirred at 70 ° C for 12 hours. After the reaction was completed, the reaction solution was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%) to obtain compound 9-4A. LCMS: m / z=530.2[M+1] + .

[0589] Step 5

[0590] Compound 9-4A (5.4 g, 10.20 mmol) was dissolved in tetrahydrofuran (50 mL), cooled to 0°C, and then sodium bicarbonate (1.03 g, 12.28 mmol) and silver trifluoromethanesulfonate (3.15 g, 12.25 mmol) were added, and then a solution of iodine (2.33 g, 9.18 mmol) in tetrahydrofuran (5 mL) was added dropwise. The reaction solution was stirred at 0°C for 15 minutes. After the reaction was completed, the reaction solution was quenched by adding a saturated sodium sulfite solution (100 mL) at 0°C, and the resulting solution was extracted with ethyl acetate (100 mL*3), and the organic phases were combined and washed once with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-40%) to obtain compound 9-5A. LCMS: m / z=656.1[M+1] + .

[0591] Step 6

[0592] Compound 9-5A (3.2 g, 4.88 mmol) was dissolved in tetrahydrofuran (3 mL) and water (1 mL), and lithium hydroxide monohydrate (615 mg, 15.64 mmol) was added at 0°C. The reaction solution was stirred at 25°C for 12 hours. After the reaction was completed, the reaction solution was adjusted to a pH of about 6 by adding saturated citric acid solution, and the resulting solution was extracted with ethyl acetate (100 mL*3). The organic phases were combined and washed once with saturated brine (100 mL), dried with anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated to obtain a crude product, which was purified by high performance liquid chromatography (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile increased from 30% to 60% in 8 minutes). The target separation solution was concentrated under reduced pressure, and then the pH value was adjusted to 7-8 with dilute ammonia water, extracted with ethyl acetate (30 mL*3), and the organic phases were combined and concentrated to obtain compound 9-6A. LCMS: m / z = 600.1 [M+1] + .

[0593] Step 7

[0594] Compound 9-6A (0.32 g, 1.18 mmol) was dissolved in N, N-dimethylformamide (7 mL), and then N, N-diisopropylethylamine (1.44 g, 11.18 mmol), compound M9 (0.67 g, 1.12 mmol) and O-(7-azabenzotriazole-1-yl)-N, N, N, N-tetramethyluronium hexafluorophosphonate (510 mg, 1.44 mmol) were added. The reaction solution was stirred at 25 ° C for 2 hours. After the reaction was completed, the reaction solution was diluted with water (50 mL), the solution was extracted with ethyl acetate (50 mL*3), the organic phase was combined and washed once with saturated brine (50 mL), dried with anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-60%) to obtain compound 9-7A. LCMS: m / z = 738.2 [M+1] + .

[0595] Step 8

[0596] 9-7A (0.63 g, 854.07 μmol) was dissolved in tetrahydrofuran (6 mL) and methanol (6 mL), and then a solution of lithium hydroxide monohydrate (0.18 g, 4.29 mmol) in water (6 mL) was added dropwise at 0°C. The reaction solution was stirred at 0°C for 1 hour. After the reaction was completed, the reaction solution was adjusted to a pH of about 6 by adding a saturated citric acid solution, and the solution was extracted with ethyl acetate (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to obtain compound 9-8A. LCMS: m / z=724.1[M+1] + .

[0597] Step 9

[0598] Compound 9-8A (0.2 g, 276.39 μmol) was dissolved in acetonitrile (20 mL) and N, N-dimethylformamide (2 mL), and then N-methylimidazole (1.13 g, 13.82 mmol) and N, N, N, N-tetramethylchloroformamidine hexafluorophosphate (388 mg, 1.48 mmol) were added. The reaction solution was stirred at 80 ° C for 2 hours. After the reaction was completed, the reaction solution was directly concentrated to remove acetonitrile, and then diluted with water (30 mL), and the solution was extracted with ethyl acetate (30 mL*3). The organic phase was combined and washed once with saturated brine (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-40%) to obtain compound 9-9A. LCMS: m / z=706.1[M+1]+ .

[0599] Step 10

[0600] Compound 9-9A (0.1 g, 141.72 μmol), potassium acetate (48.68 mg, 496.03 μmol), 2-dicyclohexylphosphine-2,6-dimethoxybiphenyl (30 mg, 71.86 μmol), tris(dibenzylideneacetone)dipalladium (26 mg, 28.54 μmol) were dissolved in toluene (5 mL), and pinacol borane (145.10 mg, 1.13 mmol) was added dropwise at 0°C under nitrogen protection. The reaction solution was stirred at 60°C under nitrogen protection for 3 hours. After the reaction was completed, the reaction solution was quenched by adding saturated ammonium chloride (10 mL) dropwise, and the solution was extracted with ethyl acetate (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-40%) to obtain compound 9-10A. LCMS: m / z=706.4[M+1] + .

[0601] Step 11

[0602] Compound 9-10A (0.1 g, 141.71 μmol), M10 (76 mg, 212.76 μmol), potassium carbonate (58.76 mg, 425.13 μmol), 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (21 mg, 28.54 μmol) were dissolved in toluene (3 mL), dioxane (1 mL) and water (1 mL), and the nitrogen was replaced 3 times. The reaction solution was stirred at 65 ° C for 12 hours. After the reaction was completed, the reaction solution was directly concentrated, and the crude product of 9-11A was directly used in the next step. LCMS: m / z = 855.6 [M + 1] + .

[0603] Step 12

[0604] Compound 9-11A (0.1 g, 116.95 μmol) and cesium carbonate (115 mg, 351.85 μmol) were dissolved in N, N-dimethylformamide (3 mL), and iodoethane (28 mg, 176.43 μmol) was added dropwise at 0°C. The reaction solution was stirred at 25°C for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated and purified by high performance liquid chromatography (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile increased from 22% to 52% in 8 minutes) to obtain the trifluoroacetate salt of compound 9-12A. LCMS: m / z=883.4[M+1] + .

[0605] Step 13

[0606] Compound 9-12A (30 mg, 33.97 μmol) was dissolved in hydrochloric acid / dioxane (2M, 2 mL), and the reaction solution was stirred at 25°C for 1 hour. After the reaction was completed, the reaction solution was directly concentrated, and the crude product 9-13A was directly used in the next step. LCMS: m / z=783.3[M+1] + .

[0607] Step 14

[0608] Compound 9-13A (0.03 g, 38.31 μmol) and (1S, 2S)-2-methylcyclopropane-1-carboxylic acid (8 mg, 79.91 μmol) were dissolved in N, N-dimethylformamide (2 mL), and then N, N-diisopropylethylamine (50 mg, 386.87 μmol) and O-(7-azabenzotriazole-1-yl)-N, N, N, N-tetramethyluronium hexafluorophosphonate (30 mg, 78.90 μmol) were added. The reaction solution was stirred at 25°C for 12 hours. After the reaction was completed, the reaction solution was filtered, and the filtrate was separated and purified by high performance liquid chromatography (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile increased from 17% to 47% in 8 minutes) to obtain the trifluoroacetate of compound 9A. LCMS: m / z = 865.3 [M+1] + . 1 HNMR(400MHz,CD3OD)δppm 8.88(s,1H),8.57(s,1H),8.20(s,1H),7.74(s,1H),7.54(s,2H),6.8-6. 41(m,2H),4.54-4.45(m,1H),4.29-4.11(m,9H),4.03-3.89(s,3H),3.77- 3.57(m,6H),3.06-2.97(m,1H),2.44-2.35(m,1H),2.27-2.15(m,5H),2.0 9-1.95(m,10H),1.66-1.57(m,5H),1.13-1.06(m,3H),0.92-0.96(m,6H).

[0609] Example 10

[0610]

[0611]

[0612] Step 1

[0613] Compound 3-4 (1 g, 1.14 mmol) and cesium carbonate (2.61 g, 8.01 mmol) were dissolved in 1-methyl-2-pyrrolidone (10 mL), cooled to 0°C, and then trifluoroethyl trifluoromethanesulfonate (2.66 g, 11.44 mmol) was added dropwise. The reaction solution was stirred at 25°C for 48 hours. After the reaction was completed, water (100 mL) was added to dilute the reaction, and the solution was extracted with ethyl acetate (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%) to obtain compound 10-1A. LCMS: m / z=955.3[M+1] + .

[0614] Step 2

[0615] At 0°C, tetrabutylammonium fluoride (1M tetrahydrofuran solution, 5 mL) was added to compound 10-1A (0.5 g, 523.01 μmol). The reaction solution was stirred at 40°C for 12 hours. After the reaction was completed, water (50 mL) was added to dilute the reaction, and the solution was extracted with ethyl acetate (50 mL*3). The organic phases were combined and washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-80%) to obtain compound 10-2A (LC-MS analysis method: 5-95AB_1.5 min, compound 10-2A retention time is 1.009 min, and its isomer retention time is 0.981 min). LCMS: m / z=717.1[M+1] + .

[0616] Steps 3 to 11

[0617] Referring to the synthesis methods of Examples 6 to 9, a reaction solution of Compound 10A was obtained, the reaction solution was filtered, and the filtrate was separated and purified by high performance liquid chromatography (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile increased from 20% to 50% in 8 minutes) to obtain the trifluoroacetate salt of Example 10A. LCMS: m / z=891.5[M+1] + . 1H NMR(400MHz,CD3OD)δppm 8.57-8.43(m,2H),7.80-7.71(m,1H),7.64(s,1H),7.59-7.48(m,2H),5.54-5.49( m,1H),5.38-5.34(m,2H),4.12-4.00(m,1H),3.70-3.68(m,1H),3.62(s,1H),3.37- 3.35(m,3H),3.05-2.99(m,4H),2.25-2.18(m,4H),2.11-2.01(m,4H),1.71-1.56(m ,5H),1.49-1.42(m,5H),1.41-1.36(m,10H),1.22-1.13(m,5H),1.12-1.04(m,3H).

[0618] Embodiment 11

[0619]

[0620] Referring to the synthesis methods of Examples 6 to 9, a reaction solution of Compound 11A was obtained. Water was added to the reaction solution until no solid precipitated, and the solid was collected by suction filtration, and then purified by high performance liquid chromatography (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile increased from 15% to 45% in 8 minutes) to obtain the trifluoroacetate salt of Compound 11A. LCMS: m / z=825.4[M+1] +. 1H NMR (400MHz, CDCl3) δ=8.92-8.78(m,1H),8.28-8.21(m,1H),7.62-7.56(m,1H),7.44-7.37(m,2H),7.36-7.31(m,1H),7.11-7.04(m,1H),5. 43-5.31(m,1H),4.97-4.86(m,1H),4.80-4.71(m,1H),4.41-4.32(m, 1H),4.23-4.13(m,2H),3.73-3.66(m,5H),3.59-3.55(m,2H),3.41(br s,4H),3.21(br dd,J=11.8,12.8Hz,3H),2.96-2.87(m,1H),2.81-2.72(m,1H),2.65-2.56 (m,1H),2.47-2.34(m,2H),2.13-2.02(m,1H),1.78-1.67(m,1H),1.48(br d,J=5.5Hz,3H),1.44-1.36(m,1H),1.31-1.19(m,3H),1.13(br d,J=5.8Hz,3H),1.02-0.93(m,6H),0.73-0.64(m,1H),0.49-0.34(m,3H).

[0621] Example 12

[0622]

[0623] Referring to the synthesis methods of Examples 6 to 9, a reaction solution of Compound 12A was obtained. Water was added to the reaction solution until no solid precipitated, and the solid was collected by suction filtration, and then purified by high performance liquid chromatography (column: C18 100×40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile increased from 17% to 47% in 8 minutes) to obtain the trifluoroacetate salt of the target compound 12A. LCMS: m / z=862.2[M+23] + . 11H NMR (400 MHz, CDCl3) δ = 8.64 - 8.48 (m, 1H), 8.32 - 8.19 (m, 1H), 7.52 (br d, J = 8.8 Hz, 1H), 7.29 (br d, J = 8.5 Hz, 1H), 7.25 - 7.22 (m, 1H), 7.17 - 7.13 (m, 1H), 6.81 - 6.73 (m, 1H), 5.45 - 5.35 (m, 1H), 4.78 (br s, 1H), 4.70 - 4.63 (m, 1H), 4.24 (brd, J = 6.0 Hz, 1H), 4.15 - 4.08 (m, 2H), 3.62 (br d, J = 10.8 Hz, 3H), 3.51 (br d, J = 11.3 Hz, 3H), 3.46 (br d, J = 5.8 Hz, 1H), 3.31 (s, 3H), 3.16 - 3.05 (m, 3H), 2.84 - 2.77 (m, 1H), 2.71 - 2.63 (m, 1H), 2.55 - 2.47 (m, 1H), 2.36 - 2.29 (m, 2H), 2.22 - 2.11 (m, 2H), 2.04 - 1.91 (m, 3H), 1.67 - 1.53 (m, 3H), 1.51 - 1.41 (m, 2H), 1.37 (br d, J = 5.8 Hz, 4H), 1.07 (br d, J = 6.0 Hz, 3H), 1.03 (br d, J = 6.0 Hz, 3H), 0.85 - 0.79 (m, 3H), 0.33 (br s, 3H).

[0624] Example 13

[0625]

[0626] Step 1

[0627] Compound M11 (1.36 g, 4.90 mmol) was dissolved in tetrahydrofuran (40 mL) and water (10 mL), and sodium bicarbonate (1.44 g, 17.14 mmol) and Boc2O (1.17 g, 5.34 mmol) were added and stirred at 25°C for 12 hours. After the reaction was completed, it was extracted with water (50 mL) and ethyl acetate (50 mL*3). The organic phase was dried and concentrated, and then purified by SFC preparation (column: DAICEL CHIRALPAK IG (250 mm*30 mm, 10 um); mobile phase: [phase A: carbon dioxide supercritical fluid, phase B: isopropanol containing 0.1% ammonia water]; phase B ratio 30% isocratic elution) to obtain compound 13-1A (Rt=3.625; SFC analysis method: column: DAICELCHIRALPAK IG (100*4.6 mm ID, 3um); mobile phase: [Phase A: carbon dioxide supercritical fluid, Phase B: isopropanol containing 0.05% ammonia]; Phase B increased from 5% to 40% in 4.5 minutes, then maintained at 5% for 1.5 minutes, flow rate 2.5mL / min, column temperature: 40°C) and 13-1B (Rt = 4.200; SFC analysis method: column: DAICEL CHIRALPAK IG (100*4.6mm ID, 3um); mobile phase: [Phase A: carbon dioxide supercritical fluid, Phase B: isopropanol containing 0.05% ammonia]; Phase B increased from 5% to 40% in 4.5 minutes, then maintained at 5% for 1.5 minutes, flow rate 2.5mL / min, column temperature: 40°C). LCMS: m / z = 321.1, 323.1 [M+1-56] +.

[0628] Step 2

[0629] Compound 3-6A (1.2 g, 1.81 mmol) and bis-naphthalene alcohol borate (688.76 mg, 2.71 mmol) were added to toluene (10 mL), and potassium acetate (354.91 mg, 3.62 mmol) was added. After nitrogen replacement, 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (132.31 mg, 180.82 μmol) was added. The reaction solution was stirred under nitrogen at 70°C for 12 hours. The reaction solution was filtered and the filtrate was concentrated. The crude product was purified by column chromatography (0-15% methanol / dichloromethane) to obtain compound 13-2A. LCMS: m / z=711.4[M+1] + .

[0630] Step 3

[0631] Compound 13-1A (0.05 g, 132.54 μmol) was dissolved in dichloromethane (2 mL), and then 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (30.49 mg, 159.04 μmol), 1-hydroxybenzotriazole (21.49 mg, 159.04 μmol), N-methylmorpholine (26.81 mg, 265.07 μmol) were added, and stirred at 0°C for 0.5 hours, and then trifluoroacetate of compound M9 (35.81 mg, 132.54 μmol) was added at 0°C, and stirred at 25°C for 1 hour. After the reaction was completed, it was extracted with water (50 mL) and ethyl acetate (50 mL*3), and the organic phase was dried and concentrated to obtain compound 13-3A. LCMS: m / z=515.0,517.0[M+1] + .

[0632] Step 4

[0633] Compound 13-3A (52.00 mg, 100.89 μmol) and compound 13-2A ​​(89.63 mg, 126.11 μmol) were dissolved in dioxane (1 mL), water (1 mL) and toluene (3 mL), and then potassium phosphate (64.25 mg, 302.67 μmol) and 1,1-di(tert-butylphosphino)ferrocenepalladium chloride (13.15 mg, 20.18 μmol) were added, and stirred at 70°C for 12 hours under nitrogen protection. After the reaction was completed, water (10 mL) was added, and then extracted with ethyl acetate (10 mL*3). After the organic phase was dried and concentrated, the crude product was separated by thin layer chromatography preparation plate (petroleum ether: ethyl acetate = 1:1) to obtain compound 13-4A. LCMS: m / z = 510.0 [M / 2+1] + .

[0634] Step 5

[0635] Referring to the synthesis methods of Examples 6 to 9, a reaction solution of Compound 13A was obtained. Water (10 mL) was added to the reaction solution, and the mixture was extracted with ethyl acetate (10 mL*3). After the organic phase was dried and concentrated, the crude product was separated by a thin layer chromatography preparative plate (dichloromethane: methanol = 10:1) to obtain Compound 13A. LCMS: m / z = 850.0 [M+1] + . 1H NMR(400MHz,CD3OD)δppm0.59(s,3H)0.67(s,3H)0.99(s,3H)1.06(br t,J=7.15Hz,6H)1.50(br s,2H)1.59(br d,J=9.29Hz,1H)1.94(s,2H)2.10(t,J=7.53Hz,1H)2.19-2.28(m,1H)2.40(br d,J=5.02Hz,2H)2.59-2.64(m,1H)2.73-2.77(m,3H)2.80(s,1H)2.95(d,J=7.28Hz,1H)3.03(s,3H)3.09-3.17(m,8H)3.43(br d,J=9.03Hz,3H)3.58-3.67(m,7H)3.87(s,1H)7.36(s,1H)7.38-7.42(m,2H)7.59(d,J=8.78Hz,1H)8.34-8.38(m,2H).

[0636] Embodiment 14

[0637]

[0638] Using compound 13-1B as raw material and referring to the synthesis method of Example 13, compound 14A was prepared.

[0639] LCMS: m / z = 850.0 [M+1] + . 1 H NMR(400MHz,CD3OD)δppm 0.56(br s,3H)0.66(br s,1H)0.90

[0640] (s,3H)1.04(br s,3H)1.13(br d,J=5.77Hz,4H)1.24-1.29(m,1H)1.31(br s,2H)1.33-1.38(m,2H)1.44(br d,J=6.02Hz,3H)1.61(br s,1H)1.73-1.79(m,1H)2.23(s,1H)2.38(s,3H)2.47(br d,J=10.04Hz,1H)2.60(br s,1H)2.67(br s,4H)2.74(br s,1H)2.97(s,1H)3.27(s,3H)3.37(br s,4H)3.60(br d,J=10.79Hz,1H)3.74(br d,J=12.55Hz,1H)4.24(br d,J=6.53Hz,1H)4.58(br s,5H)4.68(br s,1H)7.37(s,1H)7.46-7.50(m,2H)7.66(br d,J=8.53Hz,1H)8.43(s,2H).

[0641] Embodiment 15

[0642]

[0643] Step 1

[0644] Compound M10-2 (728.03 mg, 2.13 mmol) and compound 15-1A (570 mg, 2.55 mmol) were dissolved in 1,4-dioxane (10 mL) and water (2 mL), and potassium carbonate (735.57 mg, 5.32 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (155.77 mg, 212.89 μmol) were added, and the reaction solution was stirred at 50°C under nitrogen for 2 hours. The reaction solution was then filtered to remove insoluble impurities, and the filtrate was concentrated to dryness to obtain a crude product. The crude product was purified by column chromatography (0-5% methanol / dichloromethane) to obtain compound 15-2A. LCMS: m / z=312.9[M+1] + .

[0645] Step 2

[0646] Compound 15-2A (580 mg, 1.86 mmol) and neopentyl glycol diboronate (505.16 mg, 2.24 mmol) were dissolved in toluene (10 mL), and potassium acetate (457.26 mg, 4.66 mmol) and 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (136.37 mg, 186.37 μmol) were added, and the reaction solution was stirred at 80° C. under nitrogen for 2 hours. The reaction solution was then filtered to remove insoluble impurities, and the filtrate was concentrated to dryness to obtain compound 15-3A. LCMS: m / z=345.1[M+1] + .

[0647] Step 3

[0648] Compound 15-3A (640 mg, 1.86 mmol, 1 eq) and compound M5 (1.20 g, 1.86 mmol, 1 eq) were dissolved in 1,4-dioxane (20 mL) and water (4 mL), potassium carbonate (770.83 mg, 5.58 mmol) was added, nitrogen was replaced three times, 1,1-bis(diphenylphosphino)ferrocenepalladium chloride (136.03 mg, 185.91 μmol) was added, and the reaction solution was stirred under nitrogen at 70°C for 3 hours. After the reaction was completed, the reaction solution was filtered to remove insoluble impurities, and the organic solvent was removed by distillation under reduced pressure to obtain a crude product, which was purified by column chromatography (0-10% methanol / dichloromethane) to obtain compound 15-4A. LCMS: m / z=750.2[M+1] + .

[0649] Step 4

[0650] Compound 15A was prepared by referring to the synthetic methods of Examples 6 to 9. LCMS: m / z=834.5[M+H] + .

[0651] Example 16

[0652]

[0653]

[0654] Referring to the synthesis method of Examples 6 to 9, compound 16A (LCMS: m / z=820.5 [M+H] + ), compound 17A (LCMS: m / z=836.0 [M+H] + ), compound 18A (LCMS: m / z=848.5 [M+H] + ) and compound 19A (LCMS: m / z=834.5 [M+H] + ).

[0655] Biological test data

[0656] Experimental Example 1: In vitro AsPC-1 cell proliferation assay

[0657] Experimental Materials:

[0658] RPMI1640 culture medium, penicillin / streptomycin antibiotics were purchased from Gibco, fetal bovine serum was purchased from Hyclone, 3DCellTiter-Glo (cell viability chemiluminescence detection reagent) reagent was purchased from Promega, AsPC-1 cell line was purchased from ATCC, Envision multi-label analyzer (PerkinElmer).

[0659] Experimental methods:

[0660] AsPC-1 cells were seeded in ultra-low attachment 96-well U-shaped plates, with 80 μL of cell suspension per well, containing 1000 AsPC-1 cells. The cell plates were placed in a carbon dioxide incubator for overnight culture.

[0661] The compound to be tested was diluted 5-fold into 8 concentrations using a dispenser, i.e., from 2mM to 25.6nM, and a double-well experiment was set up. 78μL of culture medium was added to the middle plate, and then 2μL of the gradient diluted compound per well was transferred to the middle plate according to the corresponding position, and 20μL of each well was transferred to the cell plate after mixing. The concentration range of the compound transferred to the cell plate was 10μM to 0.128nM. The cell plate was placed in a carbon dioxide incubator and cultured for 10 days. Prepare another cell plate and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis.

[0662] Add 100 μL of cell viability chemiluminescent detection reagent to the cell plate and incubate at room temperature for 30 minutes to stabilize the luminescent signal. Read using a multi-label analyzer.

[0663] Data Analysis:

[0664] The raw data were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value of can be obtained by curve fitting with four parameters (obtained in "log (inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Table 1 provides the inhibitory activity of the compounds of the present invention on AsPC-1 cell proliferation.

[0665] Table 1: Results of in vitro screening test of compounds of the present invention

[0666] Compound No. <![CDATA[AsPC-1IC 50 (nM)]]> Compound 6A 1

[0667] Conclusion: The compounds of the present invention have significant inhibitory activity on the proliferation of AsPC-1 cells.

[0668] Experimental Example 2: In vitro cell proliferation assay

[0669] Experimental Materials:

[0670] RPMI1640 medium, DMEM medium, Ham's F12 medium, F12K medium, IMDM medium and penicillin / streptomycin antibiotics were purchased from Gibco, fetal bovine serum was purchased from Hyclone, and Envision multi-label analyzer was purchased from PerkinElmer.

[0671] 3D CellTiter-Glo (cell viability chemiluminescent detection reagent) reagent was purchased from Promega; GP2D cell line (DMEM + 10% FBS + 1% penicillin / streptomycin) was purchased from ECACC; PK-59 cell line (DMEM + 10% FBS + 1% penicillin / streptomycin), LOVO cells (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), NCI-H727 cells (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), A427 cell line (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), Capan-1 cells (IMDM + 20% FBS + 1% penicillin / streptomycin) were purchased from Nanjing Kebai Biotechnology Co., Ltd. A375 cell line (DMEM + 10% FBS + 1% penicillin / streptomycin), AsPC-1 cells (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), PSN-1 cell line (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), SW620 cells (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), HCT116 cells (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), A549 cells (F12K + 10% FBS + 1% penicillin / streptomycin), NCI-H441 cells (RPMI1640 + 10% FBS + 1% penicillin / streptomycin) were purchased from ATCC; RKN cell line (Ham's F12+10% FBS+1% penicillin / streptomycin), LU99 cell line (RPMI1640+10% FBS+1% penicillin / streptomycin) were purchased from JCRB.

[0672] Experimental methods:

[0673] The cells were seeded in an ultra-low attachment 96-well U-shaped plate, with 80 μL of cell suspension per well, containing 1000 cells. The cell plate was placed in a carbon dioxide incubator for overnight culture.

[0674] The compound to be tested was diluted 5-fold into 8 concentrations using a dispenser, i.e., from 2mM to 25.6nM, and a double-well experiment was set up. 78μL of culture medium was added to the middle plate, and then 2μL of the gradient diluted compound per well was transferred to the middle plate according to the corresponding position. After mixing, 20μL of each well was transferred to the cell plate. The concentration range of the compound transferred to the cell plate was 10μM to 0.128nM. The cell plate was placed in a carbon dioxide incubator and cultured for 5 days. Prepare another cell plate and read the signal value on the day of drug addition as the maximum value (Max value in the equation below) for data analysis.

[0675] Add 50 μL of cell viability chemiluminescent detection reagent to each well of the cell plate and incubate at room temperature for 30 minutes to stabilize the luminescent signal. Read using a multi-label analyzer.

[0676] Data Analysis:

[0677] The raw data were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by curve fitting with four parameters (obtained in "log (inhibitor) vs. response--Variable slope" mode in GraphPad Prism). Table 2 provides the experimental results of the inhibitory activity of the compounds of the present invention on cell proliferation.

[0678] Table 2: Results of in vitro cell proliferation inhibition activity test of the compounds of the present invention

[0679]

[0680]

[0681] Conclusion: The compounds of the present invention have significant inhibitory activity on the cell proliferation of RAS mutant cell lines (such as GP2D, PK-59, AsPC-1, PSN-1, RKN, Capan-1, SW620, HCT116, LOVO, A549, H441, H727, LU99 and A427), but do not show obvious inhibitory effect in wild-type independent cell lines (such as A375), and have good selectivity.

[0682] Experimental Example 3: Detection of p-ERK levels in AsPC-1 cells

[0683] Experimental Materials:

[0684] AsPC-1 cells were purchased from ATCC; RPMI-1640 medium was purchased from GIbco; fetal bovine serum was purchased from Hyclone; Advanced Phospho-ERK1 / 2 (THR202 / TYR204) KIT was purchased from Bioauxilium-Advanced Phospho; ERK1 / 2 (THR202 / TYR204) KIT components include: Advanced PhosphoERK1 / 2Eu Cryptate antibody, Advanced PhosphoERK1 / 2d2 antibody, Blocking reagent (stock solution 100X), Lysis buffer #1 (stock solution 4X), Detection buffer (ready-to-use), and the storage temperature was ≤ -16°C.

[0685] Experimental methods:

[0686] (1) Cells were seeded in a 384-well cell culture plate with a white bottom, 8 μL of cell suspension per well, and each well contained 7500 cells. The cell plate was placed in a carbon dioxide incubator and incubated at 37 degrees overnight;

[0687] (2) Dilute the test compound with 100% DMSO to 3mM as the first concentration, and then use a pipette to dilute it to 10 concentrations of 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1μM. Take 2μL of the compound and add it to 198μL of cell starvation medium. After mixing, take 15μL of the compound solution and add it to 35μL of cell starvation medium and mix. Then, add 4μL of the compound solution in the last step to the corresponding cell plate wells per well. Put the cell plate back into the carbon dioxide incubator and continue to incubate for 3 hours. At this time, the compound concentrations are 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1nM;

[0688] (3) After the incubation, add 3 μL of 5X cell lysis buffer to each well and incubate at room temperature with shaking for 30 minutes;

[0689] (4) Use detection buffer to dilute Phospho-ERK1 / 2Eu Cryptate antibody and Phospho-ERK1 / 2d2 antibody 20 times, mix them at a ratio of 1:1, add 5 μL per well to the cell culture plate, and incubate at room temperature for 2 hours;

[0690] (5) After incubation, use a multi-label analyzer to read HTRF excitation: 320nm, emission: 615nm, 665nm.

[0691] Data Analysis:

[0692] The raw data were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by curve fitting with four parameters (obtained by log(inhibitor) vs.response--Variable slope mode in GraphPad Prism). Table 4 provides the inhibitory effect of the compounds of the present invention on p-ERK. Max well: the reading value of the positive control well is 1X lysate; Min well: the reading value of the negative control well is 0.5% DMSO cell well cell lysate. The experimental results are shown in Table 3.

[0693] Table 3: Results of in vitro pERK inhibition screening test of compounds of the present invention

[0694] Compound No. <![CDATA[AsPC-1pERK IC 50 (nM)]]> Compound 6A 0.3

[0695] Conclusion: The compounds of the present invention have significant inhibitory activity on the pERK level of AsPC-1 cells.

[0696] Experimental Example 4: Detection of p-ERK levels in GP2D cells

[0697] Experimental Materials:

[0698] GP2D cell line (DMEM+10% FBS+1% penicillin / streptomycin) was purchased from ECACC; RPMI-1640 medium was purchased from GIbco; fetal bovine serum was purchased from Hyclone; Advanced Phospho-ERK1 / 2 (THR202 / TYR204) KIT was purchased from Bioauxilium-Advanced Phospho; ERK1 / 2 (THR202 / TYR204) KIT components: Advanced PhosphoERK1 / 2Eu Cryptate antibody, Advanced PhosphoERK1 / 2d2 antibody, Blocking reagent (stock solution 100X), Lysis buffer #1 (stock solution 4X), Detection buffer (ready-to-use), storage temperature is ≤-16°C.

[0699] Experimental methods:

[0700] (1) Cells were seeded in a 384-well cell culture plate with a white bottom, 8 μL of cell suspension per well, and each well contained 7500 cells. The cell plate was placed in a carbon dioxide incubator and incubated at 37 degrees overnight;

[0701] (2) Dilute the test compound with 100% DMSO to 3mM as the first concentration, and then use a pipette to dilute it to 10 concentrations of 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1μM. Take 2μL of the compound and add it to 198μL of cell starvation medium. After mixing, take 15μL of the compound solution and add it to 35μL of cell starvation medium and mix. Then, add 4μL of the compound solution in the last step to the corresponding cell plate wells per well. Put the cell plate back into the carbon dioxide incubator and continue to incubate for 3 hours. At this time, the compound concentrations are 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, and 0.1nM;

[0702] (3) After the incubation, add 3 μL of 5X cell lysis buffer to each well and incubate at room temperature with shaking for 30 minutes;

[0703] (4) Use detection buffer to dilute Phospho-ERK1 / 2Eu Cryptate antibody and Phospho-ERK1 / 2d2 antibody 20 times, mix them at a ratio of 1:1, add 5 μL per well to the cell culture plate, and incubate at room temperature for 2 hours;

[0704] (5) After incubation, use a multi-label analyzer to read HTRF excitation: 320nm, emission: 615nm, 665nm.

[0705] Data Analysis:

[0706] The raw data were converted into inhibition rate, IC using the equation (Sample-Min) / (Max-Min)*100%. 50 The value can be obtained by curve fitting with four parameters (obtained by log(inhibitor) vs.response--Variable slope mode in GraphPad Prism). Table 4 provides the inhibitory effect of the compounds of the present invention on p-ERK. Max well: the reading value of the positive control well is 1X lysate; Min well: the reading value of the negative control well is 0.5% DMSO cell well cell lysate. The experimental results are shown in Table 4.

[0707] Table 4: Results of in vitro pERK inhibition screening test of compounds of the present invention

[0708]

[0709]

[0710] Conclusion: The compounds of the present invention have significant inhibitory activity on the pERK level in GP2D cells.

[0711] Experimental Example 5: In vivo pharmacodynamics study

[0712] Purpose:

[0713] The in vivo efficacy of the compound of the present invention in a subcutaneous xenograft tumor model of human lung bronchial benign tumor NCI-H727 cells in BALB / c nude mice was studied.

[0714] Experimental methods and steps:

[0715] Experimental animals: female BALB / c nude mice, 6-8 weeks old, weighing 18-22 g; supplier: Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0716] (1) Cell culture: Human lung bronchial benign tumor cells were cultured in monolayer in vitro in GibcoRMPI1640 medium supplemented with 10% fetal bovine serum in an incubator at 37°C and 5% CO2. The cells were routinely digested and passaged with trypsin-EDTA twice a week. When the cell saturation reached 80%-90% and the number reached the required level, the cells were harvested, counted, and inoculated.

[0717] (2) Tumor cell inoculation and grouping: 0.2 mL (2×10 6 NCI-H727 cells (with Matrigel, volume ratio of 1:1) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached about 134 mm 3 The animals were divided into groups and dosed starting at 5 pm, with 6 animals in each group.

[0718] The solvent was 5% DMSO / 10% solutol / 85% water. Control group: The solvent was administered orally twice a day, with a dose of 10 μL / g; Treatment group: The compound to be tested was dissolved in the solvent and administered orally once a day, with the dose shown in Table 4.

[0719] (3) The tumor diameter was measured with a vernier caliper twice a week and the tumor volume (V) was calculated using the formula: V = 0.5a × b 2 , where a and b are the long diameter and short diameter of the tumor, respectively. The anti-tumor efficacy of the test compound is evaluated by the tumor growth inhibition rate (TGI). The calculation formula is: TGI (%) = [1-(average tumor volume at the end of drug administration in a treatment group - average tumor volume at the beginning of drug administration in the treatment group) / (average tumor volume at the end of treatment in the solvent control group - average tumor volume at the beginning of treatment in the solvent control group)] × 100%.

[0720] Experimental results:

[0721] The body weight of mice in each treatment group was maintained well after administration. The TGI was calculated based on the average tumor volume on the 21st day after administration. The specific experimental results are shown in Table 5.

[0722] Table 5: Evaluation of the anti-tumor efficacy of the compounds of the present invention in the human lung bronchial benign tumor NCI-H727 model

[0723]

[0724]

[0725] Conclusion: The compounds of the present invention exhibited excellent tumor inhibition effects in the NCI-H727 tumor model.

[0726] Experimental Example 6: In vivo pharmacodynamics study

[0727] Purpose:

[0728] The in vivo efficacy of the compound of the present invention in a subcutaneous xenograft tumor model of human pancreatic cancer PK59 cells in BALB / c nude mice was studied.

[0729] Experimental methods and steps:

[0730] Experimental animals: female BALB / c nude mice, 6-8 weeks old, weighing 18-22 g; supplier: Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0731] (1) Cell culture: Human pancreatic cancer PK59 cells were cultured in a monolayer in vitro in Gibco RMPI1640 medium supplemented with 10% fetal bovine serum in an incubator at 37°C and 5% CO2. The cells were routinely digested and passaged with trypsin-EDTA twice a week. When the cell saturation reached 80%-90% and the number reached the required level, the cells were harvested, counted, and inoculated.

[0732] (2) Tumor cell inoculation and grouping: 0.2 mL (2×10 6 PK59 cells (with Matrigel, volume ratio of 1:1) were subcutaneously inoculated on the right back of each mouse, and the average tumor volume reached about 104.4 mm 3 The animals were divided into groups and dosed starting at 5 pm, with 6 animals in each group.

[0733] The solvent was 5% DMSO / 10% solutol / 85% water. Control group: The solvent was administered by gavage once a day, with a dose of 10 μL / g; Treatment group: The compound to be tested was dissolved in the solvent, and the compound was administered by gavage once a day, with the dose shown in Table 5.

[0734] (3) The tumor diameter was measured with a vernier caliper twice a week and the tumor volume (V) was calculated using the formula: V = 0.5a × b 2 , where a and b are the long diameter and short diameter of the tumor, respectively. The anti-tumor efficacy of the test compound is evaluated by the tumor growth inhibition rate (TGI). The calculation formula is: TGI (%) = [1-(average tumor volume at the end of drug administration in a treatment group - average tumor volume at the beginning of drug administration in the treatment group) / (average tumor volume at the end of treatment in the solvent control group - average tumor volume at the beginning of treatment in the solvent control group)] × 100%.

[0735] Experimental results:

[0736] The body weight of mice in each treatment group was maintained well after administration. The TGI was calculated based on the average tumor volume on the 22nd day after administration. The specific experimental results are shown in Table 6.

[0737] Table 6 Antitumor efficacy experimental results of the compounds of the present invention in the human pancreatic cancer PK59 model

[0738]

[0739]

[0740] Conclusion: The compounds of the present invention exhibited excellent tumor inhibition effects in the PK59 tumor model.

[0741] Experimental Example 7: In vitro determination of the concentration ratio between whole blood and plasma

[0742] Experimental steps:

[0743] (1) Collect fresh whole blood from mice and humans using a blood collection tube containing EDTAK2 anticoagulant (the number of animal individuals is required to be n≥3, and the number of human individuals is required to be n≥2). Whole blood samples should be mixed evenly before use and stored at 2-8°C or on wet ice. Use within 36 hours after whole blood collection. Blank whole blood is centrifuged at 2000×g for 15 minutes at room temperature to obtain blank plasma. Check the status of the plasma; plasma samples that show hemolysis should not be used.

[0744] (2) The whole blood sample is placed in a microhematocrit centrifuge tube and centrifuged to determine the hematocrit (the volume percentage of red blood cells in the whole blood).

[0745] (3) Diclofenac (all species), chlorthalidone (mice) or chloroquine (human) were used as control compounds.

[0746] (4) Add a 2% DMSO solution of the test compound at a final concentration of 1 μM to the blank whole blood sample, and perform three parallel treatments. The final content of the organic phase in the system should not exceed 0.5% (the content of DMSO should not exceed 0.1%). Pipette a certain volume of drug-containing whole blood sample to the sample receiving plate (three parallels), and add an equal volume of blank plasma, mix well, and obtain a T0 sample.

[0747] (5) The whole blood sample containing the test compound was incubated at 37°C with shaking for 60 minutes.

[0748] (6) After the incubation, transfer a certain volume of drug-containing whole blood sample to the sample receiving plate (three parallel plates) as T 60 -Whole blood samples.

[0749] (7) Place the remaining whole blood sample in a 37°C centrifuge at 2000×g for 15 minutes to obtain a plasma sample. Transfer a certain volume of plasma to a sample receiving plate (three replicates) as T 60 - Plasma samples.

[0750] (8) During sample processing, all samples were balanced with the matrix (i.e., the same volume of blank whole blood or blank plasma was added) and mixed evenly.

[0751] (9) Add 1 volume of pure water to the balanced sample, and then add a certain volume of stop solution containing the internal standard compound to terminate the reaction.

[0752] (10) The analyte and reference compound in the sample were determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS). The ratio of the analyte peak area to the internal standard peak area was used to express the concentration in the sample.

[0753] Experimental results:

[0754] The experimental results are shown in Table 7. Among them, K B / P represents the concentration ratio of the compound in whole blood and plasma, K E / P The concentration ratio of the compound in erythrocytes and plasma is shown in Table 1, and the recovery rate (%) is shown in Table 1.

[0755] Table 7 Distribution results of the compounds of the present invention in whole blood and plasma

[0756]

[0757]

[0758] Conclusion: Compared with human and mouse plasma, the compounds of the present invention have a higher distribution in whole blood and erythrocytes.

[0759] Experimental Example 8: Pharmacokinetic Test in Mice

[0760] Purpose:

[0761] The pharmacokinetic behavior of the compounds of the present invention was evaluated in male CD-1 (ICR) mice.

[0762] Experimental methods:

[0763] The compound to be tested was dissolved in a solvent (5% DMSO / 10% solutol / 85% water). Four male CD-1 mice were divided into two groups, with two mice in each group. The first group of mice received a single intravenous injection of the compound at a dose of 1 mg / kg. The second group of mice received a single oral gavage of the compound at a dose of 10 mg / kg. Whole blood samples were collected at 0.083 (only intravenous injection group), 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration. The concentration of the test compound in the whole blood samples was determined by LC-MS / MS method.

[0764] Experimental results:

[0765] In the experiment, all animals were well tolerated and no abnormal manifestations were observed. After intravenous injection of compound 7A, the blood clearance rate (Cl) was 6.29 mL / min / kg, the steady-state apparent distribution volume (Vd) was 2.18 L / kg, and the elimination half-life (T 1 / 2 ) is 4.29h, and the area under the whole blood concentration-time curve (AUC 0-last ) was 3160 h·nmol / mL. After intragastric administration of compound 7A, the peak time (T max ) appeared 3.0h after administration and reached the peak concentration (C max ) is 2000nmol / mL, AUC 0-last It is 17119h·nmol / mL, and the bioavailability (F) is 54.3%.

[0766] Conclusion: The compounds of the present invention have high exposure, long half-life and good pharmacokinetic properties.

[0767] Experimental Example 9: Pharmacokinetic Study in Rats

[0768] Purpose:

[0769] The pharmacokinetic behavior of the compounds of the present invention was evaluated in male SD rats.

[0770] Experimental methods:

[0771] The compound to be tested was dissolved in a solvent (5% DMSO / 10% solutol / 85% water). Four male SD rats were divided into two groups, with two rats in each group. The first group of rats was given a single intravenous bolus (iv) of the compound at a dose of 1 mg / kg. The second group of rats was given a single oral gavage (po) of the compound at a dose of 10 mg / kg. Whole blood samples were collected at 0.083 (iv bolus group only), 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration. The concentration of the test compound in the whole blood samples was determined by LC-MS / MS method.

[0772] Experimental results:

[0773] In the experiment, all animals tolerated well and no abnormal performance was observed. Specific experimental results are shown in Table 8.

[0774] Table 8 Pharmacokinetic test results of the compounds of the present invention in rats

[0775]

[0776]

[0777] Conclusion: The compounds of the present invention have high exposure, long half-life and good pharmacokinetic properties.

[0778] Experimental Example 10: Pharmacokinetic Study in Beagle Dogs

[0779] Purpose:

[0780] The pharmacokinetic behavior of the compounds of the present invention was evaluated in male beagle dogs.

[0781] Experimental methods:

[0782] The compound to be tested was dissolved in a solvent (5% DMSO / 10% solutol / 85% water). Four male beagle dogs were divided into two groups of two. Group 1 animals were given a single intravenous bolus (iv) of the compound at a dose of 1 mg / kg. Group 2 animals were given a single oral gavage (po) of the compound at a dose of 5 mg / kg. Whole blood samples were collected at 0.083 (iv bolus group only), 0.25, 0.5, 1, 2, 4, 8 and 24 hours after administration. The concentration of the test compound in the whole blood samples was determined by LC-MS / MS method.

[0783] Experimental results:

[0784] In the experiment, all animals tolerated the compound well and no abnormal performance was observed. Specific experimental results are shown in Table 9.

[0785] Table 9 Pharmacokinetic test results of the compounds of the present invention in beagle dogs

[0786]

[0787] Conclusion: The compounds of the present invention have high exposure, long half-life and good pharmacokinetic properties.

Claims

1. A compound represented by formula (III), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, L is R6 or L1 is selected from -N(R9)C(=O)-; L2 is selected from C 1-6 Alkyl, the C 1-6 The alkyl group is optionally substituted with 1, 2 or 3 R a replace; L3 is selected from -CH2- and C 3-6 Cycloalkyl; L4 is selected from a single bond and -C 1-4 Alkyl-N(R 10 )C(=O)-; Ring A is Ring B is indolyl; R1 is selected from H, F, Cl, Br, I, OH, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R h replace; R2 is selected from -O- and -NH-; R3 is selected from phenyl and 5-6 membered heteroaryl, wherein the phenyl and 5-6 membered heteroaryl are each independently optionally substituted by 1, 2 or 3 R b replace; R4 and R5 are independently selected from H, C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R c replace; R6 is selected from C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl, the C 3-6 Cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R d replace; Each R7 is independently selected from H, halogen, C 1-4 Alkyl and C 1-4 Alkoxy, the C 1-4 Alkyl and C 1-4 The alkoxy groups are each independently optionally substituted with 1, 2 or 3 R e replace; Each R8 is independently selected from H, C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-7 membered heterocycloalkyl, the C 1-4 Alkyl, C 1-4 Alkoxy, C 3-6 Cycloalkyl and 3-7 membered heterocycloalkyl are each independently optionally substituted by 1, 2 or 3 R f replace; R9, R 10 Selected from H and C 1-4 Alkyl, the C 1-4 The alkyl group is optionally substituted with 1, 2 or 3 R g replace; Each R a , each R b , each R c , each R e , each R f , each R g and each R h Each independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3; Each R d are independently selected from C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 Alkenyl, the C 1-4 Alkyl, C 3-6 Cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 The alkenyl group is optionally substituted with 1, 2 or 3 R groups, each independently; Each R is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3 and OCF3; n, p and q are independently selected from 0, 1, 2 and 3; The “3-6 membered heterocycloalkyl”, “3-7 membered heterocycloalkyl” and “5-6 membered heteroaryl” each independently contain 1 or 2 heteroatoms or heteroatom groups independently selected from —NH—, —O—, —S— and N.

2. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R6 is selected from cyclopropyl, cyclobutyl, cyclopentyl, aziridine, azetidin, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl and oxhexyl, wherein the cyclopropyl, cyclobutyl, cyclopentyl, aziridine, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl and oxhexyl are independently and optionally replaced by 1, 2 or 3 R d Substituted; or, R6 is selected from For example 3. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: L is selected from Alternatively, L is selected from For example 4. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Each R1 is independently selected from H, F, OH and CH3.

5. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Structural unit Selected from 6. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Ring B is selected from 7. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R3 is selected from phenyl, pyridyl, pyrimidinyl, thiazolyl, oxazolyl, pyrazolyl and imidazolyl, wherein the phenyl, pyridyl, pyrimidinyl, thiazolyl, oxazolyl, pyrazolyl and imidazolyl are independently optionally substituted by 1, 2 or 3 R b Substituted; or, R3 is selected from 8. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: L2 is selected from 9. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: L3 is selected from -CH2-, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl; or, L3 is selected from -CH2-, 10. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R8 is selected from H, CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, homopiperidinyl and morpholinyl, wherein CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, homopiperidinyl and morpholinyl are independently and optionally substituted by 1, 2 or 3 R f Substituted; or, R8 is selected from H, 11. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Structural unit Selected from Alternatively, the structural unit Selected from 12. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R2 is selected from -O-.

13. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R7 is selected from H, F, Cl, CH3 and CH2CH3.

14. The compound according to any one of claims 1 or 2, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R4 and R5 are independently selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl and cyclopentyl, and CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl and cyclopentyl are independently selected from 1, 2 or 3 R c Substituted; or, R4 is selected from H, R5 is selected from H, CH(CH3)2 and 15. The compound according to any one of claims 1 to 3, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Each R d are independently selected from CH3, CH2CH3, phenyl, cyclopropyl, cyclobutyl, oxetanyl, azetidinyl and The CH3, CH2CH3, phenyl, cyclopropyl, cyclobutyl, oxetanyl, azetidinyl and are optionally substituted with 1, 2 or 3 R independently; or, each R d are independently selected from CH3, 16. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein the compound is represented by formula (III-1): in, R1, R3, R4, R5, R6, R7, R8, L1, L3, L4, n, and Ring A are as defined in any one of claims 1 to 15.

17. The following compound, its stereoisomer or its pharmaceutically acceptable salt, 18. The following compound, its stereoisomer or its pharmaceutically acceptable salt,

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