Macrocyclic derivatives and uses thereof

Abstract

The application discloses a kind of macrocyclic derivatives and its application, specifically discloses the compound shown in formula (III), its stereoisomer and its pharmaceutically acceptable salt.

Description

[0001] The present application claims priority to the following:

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

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

[0004] CN2023103711659, filing date: April 07, 2023;

[0005] CN2023106857489, filing date: June 09, 2023;

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

[0007] This application is a divisional of the Chinese patent application for “Macrocyclic Derivatives and Their Applications” with the application number 202380053250.9, the filing date of September 28, 2023, and the invention name. TECHNICAL FIELD

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

[0009] RAS (including KRAS, NRAS and HRAS) is a small GTPase downstream of growth factor receptors such as EGFR, a molecular switch, and a key node of the RAS-RAF-MEK-ERK signaling pathway and the PI3K-AKT-mTOR signaling pathway, which can regulate cell proliferation, survival and other events. Mutations in RAS can lead to functional activation of the protein by disrupting the GTP hydrolysis process. Under normal physiological conditions, RAS usually exists in the form of non-activated RAS(OFF) binding to GDP; while in tumor cells with RAS mutations, RAS mainly exists in the form of activated RAS(ON) binding to GTP. Since RAS mutations are an important factor in the development of cancer, mutant RAS has become an important cancer treatment target. Although covalent inhibitors targeting KRAS G12C such as Sotorasib and Adagrasib have achieved great success in KRAS G12C mutant non-small cell lung cancer, KRAS G12D , KRAS G12V , KRAS G13Cancers of other KRAS mutation types and other RAS mutation types such as NRAS still have no effective targeted therapeutic drugs. Therefore, it is of great clinical significance to develop a broad-spectrum RAS inhibitor.

[0010] Revolution Medicines, Inc. announced a class of macrocyclic compounds (WO2020132597, WO2021091982, WO2021091967, WO2021091956), which can form a ternary complex with the chaperone CypA, RAS(ON) in vivo, the formation of the ternary complex blocks the binding of RAS downstream RAF, and then inhibits the RAS-RAF-MEK-ERK signaling pathway, and achieves the effect of anti-tumor. This class of RAS inhibitors has good inhibitory effect on different RAS mutations and RAS-dependent tumors. Based on this type of macrocyclic compound, it has great clinical application value to design and synthesize a broad-spectrum RAS inhibitor with excellent drug properties. SUMMARY

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

[0012]

[0013] L is R6or

[0014] L1is selected from -N(R9)C(=O)-;

[0015] L2is selected from C 1-6 alkyl, said C 1-6 alkyl is optionally substituted with 1, 2, or 3 R a substituents;

[0016] L3is selected from -CH2- and C 3-6 cycloalkyl;

[0017] L4is 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]heptanyl, 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]heptanyl;

[0019] Ring B is selected from 5-membered heteroaryl and 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 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 3-6 Cycloalkyl 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 Rg 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 groups are each independently optionally substituted with 1, 2 or 3 R;

[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] L3is selected from -CH2- and C 3-6 cycloalkyl;

[0040] L4is 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]heptanyl, 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]heptanyl;

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

[0043] R1is selected from H, F, Cl, Br, I, OH, C 1-4 alkyl and C 1-4 alkoxy, said C 1-4 alkyl and C 1-4 alkoxy are each independently optionally substituted with 1, 2, or 3 R h ;

[0044] R2is selected from -O- and -NH-;

[0045] R3is selected from phenyl and 5-6 membered heteroaryl, said phenyl and 5-6 membered heteroaryl are each independently optionally substituted with 1, 2, or 3 R b ;

[0046] R4and R5are each independently selected from H, C 1-4 alkyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl, said C 1-4 alkyl, C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R c ;

[0047] R6is selected from C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl, said C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R d ;

[0048] each R7is independently selected from H, halogen, C 1-4 alkyl and C 1-4 alkoxy, said C 1-4 alkyl and C 1-4 alkoxy are each independently optionally substituted with 1, 2, or 3 R e ;

[0049] each R8is independently selected from H, C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl and 3-7 membered heterocycloalkyl, said C 1-4 alkyl, C 1-4 alkoxy, C 3-6 cycloalkyl and 3-7 membered heterocycloalkyl, are each independently optionally substituted with 1, 2, or 3 R f substituents;

[0050] R9, R 10 is selected from H and C 1-4 alkyl, said C 1-4 alkyl is optionally substituted with 1, 2, or 3 R g substituents;

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

[0052] each R d is 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, said C 1-4 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl, and -C(=O)-C 2-4 alkenyl are each independently optionally substituted with 1, 2, or 3 R

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

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

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

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

[0057]

[0058] L1is selected from -N(R9)C(=0)-;

[0059] L2is selected from C 1-6 alkyl, said C 1-6 alkyl is optionally substituted with 1, 2, or 3 R a substituents;

[0060] L3is selected from -CH2-;

[0061] L4is selected from a single bond and -C 1-4 alkyl-N(R 10 )C(=0)-;

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

[0063] R1is selected from H, F, Cl, Br, I, OH, C 1-4 alkyl, and C 1-4 alkoxy, said C 1-4 alkyl, and C 1-4 alkoxy are each independently optionally substituted with 1, 2, or 3 R h substituents;

[0064] R2is selected from -0- and -NH-;

[0065] R3is selected from phenyl and 5-6 membered heteroaryl, said phenyl and 5-6 membered heteroaryl are each independently optionally substituted with 1, 2, or 3 R b substituents;

[0066] R4and R5are each independently selected from H, C 1-4 alkyl, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl, said C 1-4 alkyl, C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R c substituents;

[0067] R6is selected from C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl, said C 3-6 cycloalkyl, and 3-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R d substituents;

[0068] R7is selected from H, C 1-4 alkyl, and C 1-4 alkoxy, said C 1-4 alkyl, and C 1-4alkyl, C e substituted;

[0069] R8is selected from H, C 1-4 alkyl, C 1-4 alkyl, C 3-6 cycloalkyl and 3-7 membered heterocycloalkyl, said C 1-4 alkyl, C 1-4 alkyl, C 3-6 cycloalkyl and 3-7 membered heterocycloalkyl, said C f substituted;

[0070] R9, R 10 is selected from H and C 1-4 alkyl, said C 1-4 alkyl is optionally substituted with 1, 2 or 3 R g ; each R a , R b , R c , R e , R f , R g and R h are each independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3and OCF3;

[0071] each R d is 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, said C 1-4 alkyl, C 3-6 cycloalkyl, 3-6 membered heterocycloalkyl, phenyl, 5-6 membered heteroaryl and -C(=O)-C 2-4 alkenyl is optionally substituted with 1, 2 or 3 R;

[0072] each R is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3and OCF3;

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

[0074] said "3-6 membered heterocycloalkyl", "3-7 membered heterocycloalkyl" and "5-6 membered heteroaryl" each independently comprises 1 or 2 heteroatoms or heteroatom groups independently selected from -NH-, -O-, -S- and N.

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

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

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

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

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

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

[0081] In some embodiments of the application, each R d is independently selected from CH3, and the other variables are as defined in the application.

[0082] In some embodiments of the application, each R d is independently selected from C 1-3 alkyl, said C 1-3 alkyl is optionally substituted with 1, 2, or 3 R, and the other variables are as defined in the application.

[0083] In some embodiments of the application, each R d is independently selected from CH3and CH2CH3, said CH3and CH2CH3are each independently optionally substituted with 1, 2, or 3 F, and the other variables are as defined in the application.

[0084] In some embodiments of the application, each R d is independently selected from CH3, said CH3is optionally substituted with 1, 2, or 3 F, and the other variables are as defined in the application.

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

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

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

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

[0089] In some embodiments of the application, each R f is independently selected from H, D, F, Cl, CH3and OCH3, said CH3and OCH3are each independently optionally substituted with 1, 2 or 3 R, and the other variables are as defined in the application.

[0090] In some embodiments of the application, each R f is independently selected from H, D, F, Cl, CH3, CD3, CF3and OCH3, and the other variables are as defined in the application.

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

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

[0093] In some embodiments of the application, each R1is independently selected from H, F, OH and CH3, and the other variables are as defined in the application.

[0094] In some embodiments of the application, each R1is independently selected from H, F and CH3, and the other variables are as defined in the application.

[0095] In some embodiments of the application, the structural unit is selected from and the other variables are as defined in the application.

[0096] In some embodiments of the application, the structural unit is selected from and the other variables are as defined in the application.

[0097] In some embodiments of the application, the structural unit is selected from the other variables are as defined in the application.

[0098] In some embodiments of the application, the structural unit is selected from the other variables are as defined in the application.

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

[0100] In some embodiments of the application, the ring A is selected from the other variables are as defined in the application.

[0101] In some embodiments of the application, the ring A is the other variables are as defined in the application.

[0102] In some embodiments of the application, the structural unit is selected from the other variables are as defined in the application.

[0103] In some embodiments of the application, R2is selected from -O-, the other variables being as defined in the application.

[0104] In some embodiments of the application, R3is selected from 5-6 membered heteroaryl, said 5-6 membered heteroaryl being optionally substituted with 1, 2, or 3 R b , the other variables being as defined in the application.

[0105] In some embodiments of the application, R3is selected from 5 membered heteroaryl, said 5 membered heteroaryl being optionally substituted with 1, 2, or 3 R b , the other variables being as defined in the application.

[0106] In some embodiments of the application, R3is selected from thiazolyl, thienyl, oxazolyl, pyrazolyl, and imidazolyl, each independently being optionally substituted with 1, 2, or 3 R b , the other variables being as defined in the application.

[0107] In some embodiments of the application, R3is selected from thiazolyl and thienyl, the other variables being as defined in the application.

[0108] In some embodiments of the application, R3is selected from the other variables being as defined in the application.

[0109] In some embodiments of the application, R3is selected from the other variables being as defined in the application.

[0110] In some embodiments of the application, R3is selected from phenyl, pyridyl, pyrimidinyl, thiazolyl, thienyl, oxazolyl, pyrazolyl, and imidazolyl, each of which is independently optionally substituted with 1, 2, or 3 R b and other variables are as defined in the application.

[0111] In some embodiments of the application, R3is selected from phenyl, pyridyl, pyrimidinyl, thiazolyl, oxazolyl, pyrazolyl, and imidazolyl, each of which is independently optionally substituted with 1, 2, or 3 R b and other variables are as defined in the application.

[0112] In some embodiments of the application, R3is selected from and other variables are as defined in the application.

[0113] In some embodiments of the application, R3is selected from and other variables are as defined in the application.

[0114] In some embodiments of the application, R3is selected from and other variables are as defined in the application.

[0115] In some embodiments of the application, R4is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl, and cyclopentyl, each of which is independently optionally substituted with 1, 2, or 3 R c and other variables are as defined in the application.

[0116] In some embodiments of the application, R5is selected from H, CH3, CH2CH3, CH2CH2CH3, CH(CH3)2, cyclopropyl, cyclobutyl, and cyclopentyl, each of which is independently optionally substituted with 1, 2, or 3 R c and other variables are as defined in the application.

[0117] In some embodiments of the application, R4is selected from H, and R5is selected from H, CH(CH3)2, and and other variables are as defined in the application.

[0118] In some embodiments of the application, R6is selected from cyclopentyl, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl, and oxanyl, each of which is independently optionally substituted with 1, 2, or 3 R d and the other variables are as defined in the application.

[0119] In some embodiments of the application, R6is selected from cyclopropyl, cyclobutyl, cyclopentyl, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxolanyl, and oxanyl, each of which is independently optionally substituted with 1, 2, or 3 R d and the other variables are as defined in the application.

[0120] In some embodiments of the application, R6is selected from and the other variables are as defined in the application.

[0121] In some embodiments of the application, R6is selected from and the other variables are as defined in the application.

[0122] In some embodiments of the application, R6is selected from C 3-6 cycloalkyl, which is optionally substituted with 1, 2, or 3 R 3-6 and the other variables are as defined in the application. d and the other variables are as defined in the application.

[0123] In some embodiments of the application, R6is selected from cyclopropyl, which is optionally substituted with 1, 2, or 3 R d and the other variables are as defined in the application.

[0124] In some embodiments of the application, R6is selected from and the other variables are as defined in the application.

[0125] In some embodiments of the application, R6is selected from and the other variables are as defined in the application.

[0126] In some embodiments of the application, each R7is independently selected from H, F, Cl, and C 1-3 alkyl, which is optionally substituted with 1, 2, or 3 R 1-3 and the other variables are as defined in the application. e and the other variables are as defined in the application.

[0127] In some embodiments of the application, R7 is selected from H, F, CI, CH3, CH2CH3, CH2F, CHF2, CF3, CH2CF3, and CD3, and the other variables are as defined in the application. e substituted, and the other variables are as defined in the application.

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

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

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

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

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

[0133] In some embodiments of the application, each R8 is independently selected from H, C 1-3 alkyl and 5-6 membered heterocycloalkyl, said C 1-3 alkyl and 5-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R f substituted, and the other variables are as defined in the application.

[0134] In some embodiments of the application, R8 is selected from H, CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, tetrahydropyridinyl, homopiperidinyl, morpholinyl, said CH3, CH2CH3, piperazinyl, homopiperazinyl, piperidinyl, tetrahydropyridinyl, homopiperidinyl, morpholinyl, are each independently optionally substituted with 1, 2, or 3 R f substituted, and the other variables are as defined in the application.

[0135] In some embodiments of the application, R8 is selected from H, The other variables are as defined in the application.

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

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

[0138] In some embodiments of the application, R8 is selected from H, The other variables are as defined in the application.

[0139] In some embodiments of the application, R8 is selected from H, The other variables are as defined in the application.

[0140] In some embodiments of the application, the structural unit is selected from The other variables are as defined in the application.

[0141] In some embodiments of the application, the structural unit is selected from The other variables are as defined in the application.

[0142] In some embodiments of the application, the structural unit is selected from The other variables are as defined in the application.

[0143] In some embodiments of the application, the structural unit is selected from The other variables are as defined in the application.

[0144] In some embodiments of the application, the structural unit is selected from

[0145] Other variables are as defined herein.

[0146] In some embodiments of the application, the structural unit above is is selected from Other variables are as defined herein.

[0147] In some embodiments of the application, the structural unit above is is selected from Other variables are as defined herein.

[0148] In some embodiments of the application, the structural unit above is is selected from Other variables are as defined herein.

[0149] In some embodiments of the application, the structural unit above is is selected from Other variables are as defined herein.

[0150] In some embodiments of the application, the structural unit above is is selected from Other variables are as defined herein.

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

[0152] In some embodiments of the application, ring B above is selected from indolyl, and other variables are as defined herein.

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

[0154] In some embodiments of the application, L1above is selected from -N(CH3)C(=0)-, and other variables are as defined herein.

[0155] In some embodiments of the application, L2above is selected from Other variables are as defined herein.

[0156] In some embodiments of the application, L3above is selected from -CH2-, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, and other variables are as defined herein.

[0157] In some embodiments of the application, L3above is selected from -CH2-, Other variables are as defined herein.

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

[0159] In some embodiments of the application, L3 is selected from and the other variables are as defined in the application. In some embodiments of the application, L4 is selected from a single bond and -CH2-. 2- and the other variables are as defined in the application.

[0160] In some embodiments of the application, L is selected from and the other variables are as defined in the application.

[0161] In some embodiments of the application, L is selected from and the other variables are as defined in the application.

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

[0163] In some embodiments of the application, L is selected from and the other variables are as defined in the application.

[0164] In some embodiments of the application, the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof, is selected from

[0165]

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

[0167] In some embodiments of the application, the compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof, is selected from

[0168]

[0169] wherein,

[0170] structural unit is selected from

[0171] structural unit is selected from

[0172] R6is selected from C 3-6 cycloalkyl, said C 3-6 cycloalkyl is optionally substituted with 1, 2, or 3 R d substituents;

[0173] each R dare each independently selected from C 1-4 alkyl, said C 1-4 alkyl is optionally substituted with 1, 2, or 3 R;

[0174] L3, R3, R7, each R8, and each R are as defined herein.

[0175] In some embodiments of the application, the compound of formula (VI-1), (VI-2), or (VI-3), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, is selected from is selected from The other variables are as defined herein.

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

[0177]

[0178] wherein,

[0179] T1and T3are each independently selected from CH and N;

[0180] each R1is independently selected from H, F, OH, and CH3;

[0181] R3is selected from phenyl and 5-6 membered heteroaryl, each of which is independently optionally substituted with 1, 2, or 3 R b substituents;

[0182] R6is selected from C 3-6 cycloalkyl, said C 3-6 cycloalkyl is optionally substituted with 1, 2, or 3 R d substituents;

[0183] R7is selected from H and C 1-3 alkyl, said C 1-3 alkyl is optionally substituted with 1, 2, or 3 R e substituents;

[0184] each R8is independently selected from H, C 1-3 alkyl and 5-10 membered heterocycloalkyl, said C 1-3 alkyl and 5-10 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R f substituents;

[0185] L3is selected from -CH2-, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl;

[0186] each R d is independently selected from C 1-3 alkyl, said C 1-3alkyl is optionally substituted with 1, 2, or 3 R;

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

[0188] each R e is independently selected from H, D, F, and Cl;

[0189] each R f is independently selected from H, D, F, Cl, CH3, and OCH3, which CH3and OCH3are each independently optionally substituted with 1, 2, or 3 R;

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

[0191] In some embodiments of the application, compounds of the above formula (P-1) or (P-2), stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein each R1, R3, R6, R7, each R8, T1, T3, and L3are as defined in the application for formula (VI), formula (III), or formula (I).

[0192] In some embodiments of the application, compounds of the above formula (P-1) or (P-2), stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein R3is 5-6 membered heteroaryl, and the other variables are as defined in the application.

[0193] In some embodiments of the application, compounds of the above formula (P-1) or (P-2), stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein R3is thienyl and thiazolyl, and the other variables are as defined in the application.

[0194] In some embodiments of the application, compounds of the above formula (P-1) or (P-2), stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein each R8is independently selected from H, C 1-3 alkyl and 5-6 membered heterocycloalkyl, which C 1-3 alkyl and 5-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R f , and the other variables are as defined in the application.

[0195] In some embodiments of the application, compounds of the above formula (P-1) or (P-2), stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein T1is CH, T3is CH, and the other variables are as defined in the application.

[0196] In some embodiments of the application, compounds of the above formula (P-1) or (P-2), stereoisomers thereof, or pharmaceutically acceptable salts thereof, wherein L3is selected from -CH2-, and the other variables are as defined in the application.

[0197] In some embodiments of the present application, the above-mentioned compound of formula (P-1) or (P-2), stereoisomer thereof, or pharmaceutically acceptable salt thereof is selected from the group consisting of,

[0198]

[0199]

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

[0201]

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

[0203] In some embodiments of the present application, the above-mentioned compound, stereoisomer thereof, or pharmaceutically acceptable salt thereof is selected from the group consisting of,

[0204]

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

[0206] In some embodiments of the present application, the above-mentioned compound of formula (IV), stereoisomer thereof, or pharmaceutically acceptable salt thereof is selected from the group consisting of,

[0207]

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

[0209] In some embodiments of the present application, the above-mentioned compound of formula (IV-1a), (IV-1b), stereoisomer thereof, or pharmaceutically acceptable salt thereof is selected from the group consisting of,

[0210]

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

[0212] In some embodiments of the present application, the above-mentioned compound of formula (IV), (IV-1a), (IV-1b), (IV-1a-1), (IV-1a-2), (IV-1b-1), (IV-1b-2), stereoisomer thereof, or pharmaceutically acceptable salt thereof, wherein R1 is selected from the group consisting of H, F, OH and CH3; R3 is selected from the group consisting of phenyl and thiazolyl, said phenyl and thiazolyl are each independently optionally substituted by 1, 2 or 3 Rb substituted; R6is selected from cyclopropyl, which is optionally substituted with 1, 2, or 3 R d substituted; R7is selected from H, F, Cl, CH3, and CH2CH3; R8is selected from H, each R b are each independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3, and OCF3; each R d are each independently selected from CH3,

[0213] The application also includes embodiments wherein the above-mentioned variables are combined in any manner.

[0214] The application also provides the following compounds, stereoisomers thereof, or pharmaceutically acceptable salts thereof,

[0215]

[0216]

[0217]

[0218]

[0219]

[0220]

[0221] The application also includes embodiments wherein the above-mentioned variables are combined in any manner.

[0222]

[0223]

[0224]

[0225]

[0226]

[0227]

[0228]

[0229]

[0230]

[0231]

[0232]

[0233]

[0234]

[0235]

[0236]

[0237]

[0238]

[0239]

[0240] The application also provides use of the above compound, stereoisomer thereof or pharmaceutically acceptable salt thereof in preparation of a RAS inhibitor drug.

[0241] The RAS inhibitor drug described in the application is used for treating RAS mutation and RAS-dependent tumors, such as solid tumors; further, the solid tumor is pancreatic cancer, lung cancer or colorectal cancer.

[0242] The application also provides the following synthesis methods:

[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 - intermediates

[0256]

[0257] Method 8 - intermediates

[0258]

[0259] Method 9 - intermediates

[0260]

[0261] Method 10 - intermediates

[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 compound of the present application has good combination with chaperone CypA, and the combination with CypA will block the combination of RAS downstream RAF and RAS, and then inhibit the RAS-RAF-MEK-ERK signal pathway to achieve the anti-tumor effect; the compound of the present application has 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 does not exhibit obvious inhibitory effect in wild type non-dependent cell lines (such as A375), and has good selectivity; the compound of the present application also has significant inhibitory activity on the pERK level of AsPC-1 and GP2D cells; in various pharmacokinetic experiments, the compound of the present application exhibits good pharmacokinetic properties (such as high exposure and long half-life), and has higher distribution in whole blood and red blood cells relative to human and mouse plasma; in in vivo efficacy experiments, the compound of the present application exhibits excellent tumor inhibition effect, and has small dosage, high safety, and wide application prospect.

[0284] Definitions and Descriptions

[0285] Unless otherwise indicated, the following terms and phrases as used herein are intended to have the following meanings. A particular term or phrase should not be construed as undefined or unclear without a specific definition, but should be understood according to the ordinary meaning. When a trade name appears herein, it is intended to refer to its corresponding product or active ingredient thereof.

[0286] The term "pharmaceutically acceptable" as used herein with respect to compounds, materials, compositions, and / or dosage forms, means those that 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 problem or complication, commensurate with a reasonable benefit / risk ratio.

[0287] The term "pharmaceutically acceptable salt" refers to a salt of a compound of the present application, which is prepared from a compound of the present application having the specific substituents discovered herein with a relatively nontoxic acid or base. Alkali addition salts can be obtained by contacting a compound of the present application in pure solution or in a suitable inert solvent with a sufficient amount of a base. Acid addition salts can be obtained by contacting a compound of the present application in pure solution or in a suitable inert solvent with a sufficient amount of an acid. Certain specific compounds of the present application contain both basic and acidic functionalities and can be converted into either the base or acid addition salt.

[0288] The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound that contains an acid or a base moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent or in a mixture of both.

[0289] The compounds of the present application can exist in particular geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis- and trans-forms, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)-isomers, as well as racemic mixtures and other mixtures thereof, such as those that contain one or more enantiomeric or diastereomeric excess, all of which are intended to be within the scope of the present application. Additional asymmetric carbon atoms can be present in a substituent group such as an alkyl group. All such isomers, as well as mixtures thereof, are intended to be within the scope of the present application.

[0290] The term "enantiomeric" or "optical isomer" means a stereoisomer whose mirror image is not superimposable.

[0291] The term "cis" or "geometric isomer" means a stereoisomer resulting from restricted rotation about a single bond or ring atom.

[0292] The term "diastereomeric" means a stereoisomer whose molecules have two or more chiral centers and whose molecules are not mirror images of one another.

[0293] "(+)" means dextrorotary, "(-)" means levorotary, and "(±)" means racemic unless otherwise indicated.

[0294] Unless otherwise indicated, a wedged solid line bond and a wedged dashed line bond indicate the absolute configuration of a stereogenic center, a straight solid line bond and a straight dashed line bond indicate the relative configuration of a stereogenic center, a wavy line indicates a wedged solid line bond and / or a wedged dashed line bond or a wavy line indicates a straight solid line bond and / or a straight dashed line bond

[0295] Certain compounds of the present application can exist in atropisomers, which are conformational isomers that arise 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 as pure individual atropisomers, or enriched in one atropisomer, or as non-specific mixtures of each. If the rotational potential about the single bond is sufficiently high, and the interconversion between conformations is sufficiently slow, then separation of the isomers can be allowed.

[0296] The term "enriched in one isomer," "isomerically enriched," "enriched in one enantiomer," or "enantiomerically enriched," unless otherwise specified, means that the content of one isomer or enantiomer is less than 100% and that 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] The term "enantiomeric excess" or "isomeric excess," unless otherwise specified, means the difference between the relative percentages of the two isomers or enantiomers. For example, where one isomer or enantiomer is present in 90% and the other isomer or enantiomer is present in 10%, the enantiomeric or isomeric excess (the 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 application 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 cleaved to provide the pure desired enantiomer. Alternatively, where a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group) is present in the molecule, a diastereomeric salt form of the compound with an appropriate optically active acid or base can be formed, and the diastereomeric salt separated by conventional means, and the desired enantiomer recovered by treating with base or acid, as appropriate. Additionally, separation of the enantiomers and diastereomers is typically accomplished by chromatography using a chiral stationary phase, optionally in combination with chemical derivatization (e.g., formation of a carbamate from an amine). The compounds of the present application can contain unnatural proportions of atomic isotopes at one or more atoms, for example, proportion of deuterium ( 3 (¾, iodine-125 ( 125 (I), or C-14 ( 14C). For example, deuterium can be substituted for hydrogen to form deuterated drugs, which have advantages over non-deuterated drugs, such as reduced toxicity, increased drug stability, enhanced efficacy, and increased drug biohalf-life. All isotopic variations of the compounds of the present application, whether radioactive or not, are included within the scope of the present application.

[0299] The term "substituted" means that any one or more hydrogen atoms on a given atom are replaced with a substituent group, which can include isotopes of hydrogen, provided that the valency of the given atom is not exceeded and that the substitution results in a stable compound. When the substituent is oxo (i.e., =0), it means that two hydrogen atoms are replaced. The term "optionally substituted" means that the atom can or can not be substituted and that the type and number of substituents, if present, can be any that is chemically possible.

[0300] When any variable (e.g., R) occurs more than one time in a compound, its definition in each occurrence is independent of the definition of the other occurrences. Thus, for example, if a group is substituted with 0-2 R groups, then the group can optionally be substituted with up to two R groups, and at each occurrence R is selected independently.

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

[0302] When the number of occurrences of a substituent is zero, it means that the substituent is absent, such as -A-(R)0 means that the structure is actually -A.

[0303] When a substituent is null, it means that the substituent is absent, such as X is null in A-X 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 to which it is attached are directly connected, such as L represents a single bond in A-L-Z means that the structure is actually A-Z.

[0305] When the listed linking groups do not indicate the direction of attachment, the direction of attachment is arbitrary, such as where the linking group L is -M-W-, then -M-W- can either attach ring A and ring B to form or ring A and ring B to form The combination of substituents and / or variations thereof is permitted only if the combination results in a stable compound.

[0306] Unless otherwise specified, when a group has one or more points of attachment, then at each occurrence, one or more of said points of attachment can be attached to a group by a chemical bond. When the manner of attachment of the chemical bond is not specified, and a point of attachment has an H atom, then upon attachment of the chemical bond, the number of H atoms at that point of attachment is reduced by one to form a group of the corresponding valence. The chemical bond by which the point of attachment is connected to the other group can be represented by a straight, solid line a straight, dashed line or a wavy line . For example, the straight, solid line in -OCH3represents attachment to the other group through the oxygen atom in the group; the straight, dashed line in -NH2represents attachment to the other group through both ends of the nitrogen atom in the group; the wavy line in -Ph represents attachment to the other group through the carbon atoms at positions 1 and 2 in the phenyl group; represents that any point of attachment on the piperidinyl group can be attached to the other group by one chemical bond, including at least the four ways of attachment, even though H atoms are drawn on -N-, but also including groups of this type of attachment, only upon attachment of one chemical bond, the H at that point of attachment is reduced by one to form a corresponding monovalent piperidinyl group.

[0307] Unless otherwise specified, the number of atoms in a ring is defined as the number of ring members, e.g., "5-7 membered ring" means a "ring" that has 5-7 atoms arranged in a ring.

[0308] Unless otherwise specified, the term "C 1-6 alkyl" by itself or in combination with other terms refers to a straight or branched saturated carbon hydride group consisting of 1 to 6 carbon atoms. The C 1-6 alkyl group includes C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C6and C5alkyl, etc.; which can be monovalent (e.g., methyl), divalent (e.g., methylene), or multivalent (e.g., methine). Examples of C 1-6 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), 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 refers to a straight or branched saturated carbon hydride group consisting of 1 to 4 carbon atoms. The C1-4 Alkyl includes C 1-2 , C 1-3 , and C 2-3 alkyl and the like; which can be monovalent (such as methyl), divalent (such as methylene), or multivalent (such as methine). C 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, means those alkyl groups attached to the remainder of the molecule through an oxygen. The C 1-4 Alkoxy includes C 1-3 , C 1-2 , C 2-4 , C4, and C3 alkoxy and the like; which can be monovalent, divalent, or multivalent. C 1-4 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propyloxy (including n- propyloxy and isopropyloxy), butyloxy (including n-butyloxy, isobutyloxy, s-butyloxy, and t-butyloxy), and the like.

[0310] Unless otherwise specified, "C 3-6 Cycloalkyl" by itself or in combination with other terms, means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms. The C 3-6 Cycloalkyl includes monocyclic and polycyclic rings, where polycyclic rings include spiro, fused, and bridged rings. The C 3-6 Cycloalkyl includes C 3-5 , C 4-5 , and C 5-6 cycloalkyl and the like; which can be monovalent, divalent, or multivalent. C 3-6 Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, bicyclo[l. l. l]pentanyl, and the like.

[0311] Unless otherwise specified, the term "3-10 membered heterocycloalkyl" by itself or in combination with other terms, means 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, with the remainder being carbon atoms, wherein the carbon atoms are optionally oxidized (i.e., C=0), 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 group includes monocyclic and polycyclic rings, where polycyclic rings include spiro, fused, and bridged rings. Further, with respect to this "3-10 membered heterocycloalkyl" group, a heteroatom can occupy the position of attachment of the heterocycloalkyl group to the rest of the molecule. The 3-10 membered heterocycloalkyl group 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 groups, and the like. It can be monovalent, divalent, or multivalent. 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, and the like), tetrahydrofuranyl (including tetrahydrofuran-2-yl, and the like), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, and the like), piperazinyl (including 1-piperazinyl and 2-piperazinyl, and the like), morpholinyl (including 3-morpholinyl and 4-morpholinyl, and the like), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, or hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, tetrahydropyridinyl, and the like.

[0312] Unless otherwise specified, the term "3-7 membered heterocycloalkyl," by itself or in combination with other terms, represents 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, with the remainder being carbon atoms, wherein the carbon atoms are optionally oxidized (i.e., 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 group includes monocyclic and polycyclic rings, where polycyclic rings include spiro, fused, and bridged rings. Further, with respect to this "3-7 membered heterocycloalkyl" group, a heteroatom can occupy the position of attachment of the heterocycloalkyl group to the rest of the molecule. The 3-7 membered heterocycloalkyl group 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 groups, and the like. It can be monovalent, divalent, or multivalent. 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, and the like), tetrahydrofuranyl (including tetrahydrofuran-2-yl, and the like), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, and the like), piperazinyl (including 1-piperazinyl and 2-piperazinyl, and the like), morpholinyl (including 3-morpholinyl and 4-morpholinyl, and the like), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1,2-oxazinyl, 1,2-thiazinyl, or hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, tetrahydropyridinyl, and the like.​

[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 oxoed (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 group includes monocyclic and polycyclic rings, wherein the polycyclic ring includes spirocyclic, fused, and bridged rings. Furthermore, with respect to the term "3-6 membered heterocycloalkyl group," a heteroatom may occupy the position at which the heterocycloalkyl group is connected to the rest of the molecule. The 3-6 membered heterocycloalkyl group includes 4-6 membered, 5-6 membered, 4 membered, 5 membered, and 6 membered heterocycloalkyl groups. It may 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" are used interchangeably in the present invention. The term "5-6 membered heteroaryl" refers to 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., to form 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 group can be attached to the rest of the molecule by a heteroatom or carbon atom. The 5-6 membered heteroaryl group includes 5 membered and 6 membered heteroaryl groups. It can be monovalent, divalent, or multivalent. Examples of the 5-6 membered heteroaryl group 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.), thiophenyl (including 2-thiophenyl and 3-thiophenyl, 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 fused to 5-membered heteroaryl" means a 5-membered heteroaryl group fused to another 5-membered heteroaryl group through adjacent 2 atoms. Examples of "5-membered heteroaryl fused to 5-membered heteroaryl" include, but are not limited to

[0316] Unless otherwise specified, C n-n+m or C n -C n+m Any one of the specific instances including n to n+m carbons, for example C 1-12 including C1, C2, C3, C4, C5, C6, C7, C8, C9, C 10 , C 11 , and C 12 Any one of the ranges from n to n+m, for example 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 9-12and so forth. Similarly, n- to n+m-membered means the number of atoms in the ring is n to n+m, for example, 3- to 12-membered ring includes 3-membered, 4-membered, 5-membered, 6-membered, 7-membered, 8-membered, 9-membered, 10-membered, 11-membered, and 12-membered rings, and also includes any range within n to n+m, for example, 3- to 6-membered ring includes 3- to 6-membered, 3- to 9-membered, 5- to 6-membered, 5- to 7-membered, 6- to 7-membered, 6- to 8-membered, and 6- to 10-membered rings, and so forth.

[0317] The term "leaving group" refers to a functional group or atom that can be displaced by another functional group or atom through a substitution reaction, such as a nucleophilic substitution reaction. For example, representative leaving groups include triflate; chloro, bromo, iodo; sulfonate groups, such as methanesulfonate, toluenesulfonate, p-bromobenzenesulfonate, p-toluenesulfonate, and the like; acyloxy groups, such as acetoxy, trifluoroacetoxy, and the like.

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

[0319] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by a combination of the specific embodiments with other chemical synthetic methods, and equivalents thereof as appreciated by those skilled in the art, preferred embodiments including, but not limited to, the examples of the present application.

[0320] The compounds of the present application can be confirmed by routine methods known to those skilled in the art. If the present application relates to the absolute configuration of a compound, the absolute configuration can be confirmed by routine techniques in the art. For example, single crystal X-ray diffraction (SXRD), a single crystal is grown and diffracted intensity data is collected using a Bruker D8 venture diffractometer with Cu Kα radiation, scanning mode: After the relevant data is collected, the crystal structure is further resolved using direct methods (Shelxs97) to confirm the absolute configuration.

[0321] The solvents used in the present application are commercially available. The compounds are named according to the conventional naming principles in the art or using software, and the commercially available compounds are named using the supplier's catalog name. BRIEF DESCRIPTION OF DRAWINGS

[0322] Figure 1 Binding mode of compound A with CypA protein.

[0323] Figure 2 Binding mode of compound B with CypA protein.

[0324] Figure 3 Binding mode of compound C with CypA protein.

[0325] Figure 4 Binding mode of compound D with CypA protein.

[0326] Figure 5 Binding mode of compound E with CypA protein.

[0327] Figure 6 Binding mode of compound F with CypA protein.

[0328] Figure 7 Binding mode of compound G with CypA protein.

[0329] Figure 8 Binding mode of compound H with CypA protein.

[0330] Figure 9 Binding mode of compound I with CypA protein.

[0331] Figure 10 Binding mode of compound J with CypA protein. DETAILED DESCRIPTION

[0332] The application will be described in detail below with the help of examples, but this does not mean any unfavorable limitation of the application. The application has been described in detail herein, and specific embodiments thereof have also been disclosed. It will be obvious to those skilled in the art that various changes and modifications can be made to the specific embodiments of the application without departing from the spirit and scope of the application.

[0333] Computational Example 1

[0334]

[0335] The co-crystal complex of human CypA protein with natural product Sanglifehrin A (PDB ID code: 1YND) was used as a docking template in conjunction with the mode prediction. To prepare the protein, hydrogen atoms were added using Maestro [1] Protein Preparation Wizard module, and the co-crystal structure was subjected to hydrogen bond optimization, removal of water molecules other than the ligand Sanglifehrin A in the co-crystal complex, and energy optimization of the whole using OPLS4 force field. For the preparation of the ligand: the molecule to be docked was generated in 3D structure and subjected to energy minimization using LigPrep [2] . Compound A was docked into the prepared 1YND protein structure using Induced Fit Docking [3] and Protocol: Extended Sampling options in Maestro (version 2021-2). The best binding model was selected, see Figure 1 . The selected model preserved the main hydrogen bond interactions of the ligand Sanglifehrin A with the protein in the original co-crystal complex. The center of mass of Compound A in this binding model was used to generate a docking grid, and the docking model was generated using Glide [4] Receptor Grid Generation module. Based on this docking model, compounds B-J were docked using SP docking mode in Glide [3] , and the binding modes of compounds B-J are shown in Figures 2 to 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 compound of the present application has good binding with human Cyp A protein. The compound of the present application forms hydrogen bonds with Arg55, Gln63, Asn102 and His126. In addition, Arg55 forms a cationic π bond with the indole ring. Since the compound of the present application acts on the protein surface, the listed hydrogen bonds serve as anchor points, not only reproducing the binding mode of the natural product Sanglifehrin A in the co-crystal complex, but also tightly fixing the compound of the present application on the protein surface. The binding of the compound of the present application to CypA will block the binding of RAS downstream RAF to RAS, thereby inhibiting the RAS-RAF-MEK-ERK signaling pathway to achieve an anti-tumor effect.

[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 reaction solution was adjusted to pH 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 reduced pressure distillation 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), then 4- dimethylaminopyridine (7.4 g, 60.57 mmol) and acrolein (38.58 g, 688.11 mmol) were added. The reaction was stirred at 50 °C for 48 h. The reaction was cooled to room temperature, then poured into 2000 mL of water, extracted with ethyl acetate (300 mL*3), the combined organic phase was washed with dilute hydrochloric acid (2 M, 300 mL*3), then washed with water (500 mL) once, and washed with saturated brine once (200 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product. The crude product was then distilled under reduced pressure at about -0.1 MPa, 90-95 °C to give 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 was stirred at 80 °C for 16 h. The reaction was concentrated, and the crude product was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-15%) to give compound M2-3. 1 H NMR (400 MHz, CDC13) δ ppm 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 separated by SFC chiral separation (column: DAICEL CHIRALPAK IC (250 mm*50 mm, 10 um); mobile phase: A phase supercritical carbon dioxide, B phase [0.1% ammonia water in isopropyl alcohol]; B%: 20%-20%) to give compound M2-3A. SFC analysis method (column: Cellulose 2 (150 mm*4.6 mm, I.D., 5 um; mobile phase: A phase supercritical carbon dioxide, B phase [0.05% diethylamine in isopropyl alcohol]; isocratic elution B%: 5%-5%, column temperature: 35 °C, column pressure: 1500 psi), ee = 100%, the peak time of compound M2-3A was 2.798 min, and the peak time of its enantiomer was 2.133 min. LCMS: m / z = 379.0 [M+23]+.

[0353] Step 4

[0354] Compound M2-5 was prepared according to the procedure described in Example 1, Step 4, using compound M2-3A (1 g, 2.81 mmol) and trimethylsulfoxonium iodide (3.09 g, 14.03 mmol). LCMS: m / z = 371.2 [M+1] + ; 1 H NMR (400 MHz, CDC13) δ ppm 4.22-4.33 (m, 1H), 4.07-4.20 (m, 2H), 2.56-2.73 (m, 1H), 2.11-2.24 (m, 1H), 1.84-1.95 (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), then trifluoroacetic acid (3.07 g, 26.93 mmol, 2 mL) was added. The reaction was stirred at 20 °C for 2 hours. The reaction was concentrated to give the crude trifluoroacetate salt of M2-6, which was used directly in the next step.

[0357] Step 6

[0358] The crude trifluoroacetate salt of M2-6 from Step 5 was dissolved in dichloromethane (10 mL), 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), 1-hydroxybenzotriazole (39 mg, 284.8 μmol) were added. The reaction was stirred at 25 °C for 12 hours. The reaction was diluted with water (50 mL), and the solution was extracted with dichloromethane (50 mL*3). The organic phases were combined, washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The crude product was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0~60%) to give compound M2. LCMS: m / z = 503.1, 505.1 [M+1]+ .

[0359] Reference Example 3: Compound M3

[0360]

[0361] Step 1

[0362] To a solution of compound M2-3A (4.28 g, 12.01 mmol) in toluene (24 mL) was added tetrabutylammonium bromide (116.14 mg, 360.26 μmol) and difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol). The mixture was stirred at 110 °C for 4 h. Additional difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol) was added and stirred at 110 °C for 16 h. Additional difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol) was added and stirred at 110 °C for 4 h. Additional difluorobromomethyltrimethylsilane (3.66 g, 18.01 mmol) was added and stirred at 110 °C for 16 h. The toluene was concentrated under reduced pressure and the residue was added to 20 mL of water and extracted with ethyl acetate (20 mL*3). The organic phase was combined and 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 flash silica gel column chromatography (0-10% ethyl acetate / petroleum ether) to give compound M3-1. LCMS: m / z = 206.9 [M-2Boc+1] + ; 1 H NMR (400 MHz, CDCl3) δ 4.96-5.25 (m, 1H), 4.17-4.51 (m, 3H), 3.19 (br s, 1H), 2.18 (br d, J = 10.54 Hz, 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) and trifluoroacetic acid (7.68 g, 67.31 mmol, 5 mL) was added. The reaction was stirred at 25 °C for 3 h. After the reaction was completed, the organic solvent was removed by distillation under reduced pressure to give 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] Compound M1 (1.04 g, 2.95 mmol) and compound M3-2 (507 mg, trifluoroacetate crude) were dissolved in dichloromethane (10 mL) at 0 °C, N-methylmorpholine (2.49 g, 24.59 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (1.41 g, 7.38 mmol), 1-hydroxybenzotriazole (66.45 mg, 491.78 μmol) were added successively, and 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] Compound M4-1 (25 g, 136.61 mmol) in tetrahydrofuran was added to methyl magnesium bromide tetrahydrofuran solution (3 M, 91.07 mL) at 0 °C under nitrogen protection, 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 (500 mL*2), and the combined organic phase was then dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by flash silica gel column chromatography (ethyl acetate / petroleum ether 0-30%) to obtain M4-2. LCMS: m / z = 200.0, 202.0 [M+1] + ; 1 H NMR (400 MHz, CDCl3) δ 8.58 (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] To a mixture of formic acid (12.68 g, 263.96 mmol) and triethylamine (133.55 g, 1.32 mol, 183.70 mL) was added compound (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethanedi-amine (p-isopropylbenzene) chlororuthenium (700.83 mg, 1.10 mmol) at 0 °C under nitrogen protection. Then, it was stirred at 40 °C for 15 min, then cooled to 25 °C, added M4-2 (22 g, 109.98 mmol), then stirred at 40 °C for 2 h. The reaction solution was concentrated under reduced pressure, the residue was diluted with 100 mL, extracted with ethyl acetate (50 mL*2), the combined organic phase was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude product. The crude product was purified by flash silica gel column chromatography (ethyl acetate / petroleum ether 0-40%) to give compound M4-3. LCMS: m / z = 202.0, 204.0 [M+1] + ; 1 H NMR (400 MHz, CDCl3) δ 8.50 (d, J = 4.8 Hz, 1H), 7.84 (d, J = 8.0 Hz, 1H), 7.13 (dd, J = 8.0, 4.8 Hz, 1H), 5.11 (br s, 1H), 4.44 (br s, 1H) 1.44 (d, J = 4.8 Hz, 3H).

[0373] Step 3

[0374] To a solution of compound M4-3 (22 g, 108.88 mmol) in tetrahydrofuran (200 mL) was added sodium hydride (5.23 g, 130.66 mmol, 60% purity) portionwise at 0 °C under nitrogen protection. After stirring at 0 °C for 1 h, iodomethane (217.77 mmol, 13.56 mL) was added, and after the addition was complete, the mixture was slowly warmed from 0 °C to 25 °C, and stirred at this temperature for 2 h. The mixture was quenched by adding it to saturated aqueous ammonium chloride solution (100 mL) at 0 °C, water (200 mL) was added, and then ethyl acetate (100 mL*2) was added to extract, the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography (ethyl acetate: petroleum ether 0-20%) to give compound M4-4. LCMS: m / z = 216.0, 218.0 [M+1] + ; 1 H NMR (400 MHz, CDCl3) δ 8.59 (d, J = 3.2 Hz, 1H), 7.82 (dd, J = 8.4, 3.2 Hz, 1H), 7.08 (dd, J = 8.0, 4.4 Hz, 1H), 4.91 (dd, J = 13.2, 6.4 Hz, 1H), 3.29 (s, 3H), 1.44 (d, J = 4.8 Hz, 3H).

[0375] Step 4

[0376] To a solution of compound M4-4 (5 g, 23.14 mmol) in toluene (50 mL) was added bis(pinacolato)diboron (7.05 g, 27.77 mmol), Pd(dppf)Cl2(1.69 g, 2.31 mmol) and potassium acetate (4.54 g, 46.28 mmol), after addition, replaced with nitrogen three times. Stirring at 100 °C for 3 hours. Cooled to room temperature, added water (100 mL), then extracted with ethyl acetate (50 mL*2), collected organic phase, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated under reduced pressure, the crude product was purified by silica gel column separation (ethyl acetate: petroleum ether 0-25%) to obtain compound M4. LCMS: m / z = 182.0 [boric acid M+1] + ; 1 H NMR (400 MHz, CDCl3) δ 8.59 (dd, J = 4.8, 1.6 Hz, 1H), 7.90 (dd, J = 7.6, 2.0 Hz, 1H), 7.16 (dd, J = 8.4, 4.8 Hz, 1H), 4.77 (dd, J = 13.2, 6.4 Hz, 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-butyldiphenylchlorosilane (40 g, 145.53 mmol) and imidazole (12.3 g, 180.68 mmol) were dissolved in 300 mL of anhydrous, the mixture was stirred at 15 °C for 20 hours. 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, concentrated under reduced pressure, the residue was purified by flash chromatography column (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-10%) to obtain compound M5-2. 1 H NMR (400 MHz, CDCl3) δ = 7.67 (dd, J = 1.6, 8.0 Hz, 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, adjusted to pH 3-4 with concentrated hydrochloric acid, and separated. The aqueous phase was extracted with ethyl acetate (2*500 mL), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound M5-3. 1 H NMR (400 MHz, CDC13) δ = 7.58 (dd, J = 1.6, 8.0 Hz, 4H), 7.39-7.27 (m, 6H), 3.58 (s, 2H), 1.15 (s, 6H), 0.97 (s, 9H).

[0383] Step 3

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

[0385] Step 4

[0386] To a solution of compound M5-4 (51.00 g, 136.00 mmol) in 200 mL of anhydrous dichloromethane at 0 °C, tin tetrachloride (136.00 mmol, 15.92 mL) was slowly added, and the mixture was stirred under a nitrogen atmosphere for half an hour. Compound M5-5 (26.66 g, 136 mmol) in 200 mL of anhydrous dichloromethane was slowly added dropwise to the above reaction mixture at 0 °C, and the reaction system was stirred at 0 °C under a nitrogen atmosphere for 1 hour. It was quenched by adding 500 mL of water, filtered, and the filtrate was allowed to stand and separate. The aqueous phase was extracted with dichloromethane (3*500 mL), and the combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by flash column 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 (400 MHz, CDC13) δ = 8.70 (d, J = 1.6 Hz, 1H), 8.62 (br s, 1H), 7.69 (d, J = 3.2 Hz, 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] Lithium borohydride tetrahydrofuran solution (1M, 123.47 mL) was slowly added to compound M5-6 (22 g, 41.16 mmol) in 220 mL of anhydrous tetrahydrofuran at 0 °C under nitrogen atmosphere. After the addition was completed, the temperature was raised to 60 °C and stirred for 15 hours. After cooling to room temperature, 10 mL of saturated aqueous ammonium chloride solution was slowly added, followed by the addition of 100 mL of ethyl acetate, 50 mL of saturated brine. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-20%) to give 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), iodine (9.02 g, 35.54 mmol), and silver trifluoromethanesulfonate (10.04 g, 39.09 mmol) were added to 185 mL of anhydrous tetrahydrofuran successively. The mixture was stirred at 20 °C for 2 hours. 50 mL of saturated aqueous sodium sulfite solution was added to quench, and 200 mL of ethyl acetate was added to dilute. The mixture was filtered, and the filtrate was allowed to stand to separate into two phases. The organic phase was collected, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound M5. 1 H NMR (400 MHz, CDC13) δ = 8.70 (d, J = 1.6 Hz, 1H), 8.62 (br s, 1H), 7.69 (d, J = 3.2 Hz, 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).

[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 a solution of trimethylsilyldiazomethane in hexane (2 M, 78.05 mL) was added dropwise at 0 °C. Stirring was continued at 0 °C for 10 min. 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), and tert-butyl hydrazinecarboxylate (11.69 g, 70.35 mmol) was added, and the mixture was stirred at 70 °C for 12 h. Stirring was stopped, and water (100 mL) and ethyl acetate (100 mL) were added when the system cooled to room temperature. The separated organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound M6-3, which was used directly in the next step. LCMS: m / z = 201 [M + 1 - 56] + .

[0397] Step 3 Compound M6-3 (7 g, 27.31 mmol) in tetrahydrofuran (10 mL) was added dropwise slowly to borane dimethyl sulfide (10 M, 273.12 mL) at 30 °C under nitrogen protection. After stirring at 0 °C for half an hour, the temperature was raised to room temperature and stirring was continued for 1 h. Methanol (250 mL) was added dropwise slowly at 0 °C to quench the reaction, and the quenched reaction was concentrated under reduced pressure to give the crude product, which was purified by column chromatography (eluent: ethyl acetate / petroleum ether 3% to 7%) to give 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 successively at 25 °C. Stirring was continued at 25 °C for 1 h. Water (50 mL) and ethyl acetate (30 mL) were added to the reaction system, and the obtained organic phase was dried over anhydrous sodium sulfate. The filtrate was concentrated, and the obtained crude product was purified by column chromatography (eluent: ethyl acetate / petroleum ether 3% to 7%) to give 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), lithium bis(trimethylsilyl)amide (1 M, 4.90 mL) was added dropwise under nitrogen protection at -70 °C, after stirring at -70 °C for half an hour, trimethylsilyl chloride (531.94 mg, 4.90 mmol) was added dropwise, after continuing to stir at -70 °C for one hour, N-bromosuccinimide (1.16 g, 6.53 mmol) was added, and the temperature was slowly increased to 25 °C and the stirring was continued for 1 hour. Saturated brine (30 mL) was added, and extraction was performed 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 directly used in the next step. LCMS: m / z = 353, 355 [M+1-100-56] + .

[0402] Step 6

[0403] A solution of citric acid (618.66 mg, 2.94 mmol) in water (5 mL) was added to compound M6-6 (500 mg, 981.34 μmol) in tetrahydrofuran (20 mL), and stirred at 25 °C for 1 hour. After the reaction was completed, saturated brine (30 mL) was added, and extraction was performed with ethyl acetate (30 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated, and 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 heated to 60 °C and stirred for 12 hours. Filtration was performed, 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 (400 MHz, CDCl3) 5.24-4.93 (m, 1H), 4.45 (br d, J = 4.6 Hz, 1H), 3.77-3.71 (m, 3H), 2.89 (qd, J = 5.8, 11.6 Hz, 1H), 2.39 (td, J = 4.9, 10.0 Hz, 1H), 2.13 (td, J = 5.0, 10.0 Hz, 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), hydrogen chloride ethyl acetate solution (5 mL, 4 M) was added, and the mixture was 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] To a solution of M8-1 (22.5 g, 111.94 mmol) in tetrahydrofuran (250 mL) was added triethylamine (111.94 mmol, 15.58 mL) and 1-methylpiperazine (16.82 g, 167.91 mmol, 18.63 mL), and the mixture was stirred at 60 °C for 16 hours. The reaction solution was diluted with water (200 mL), and the solution was extracted with ethyl acetate (100 mL*3). The organic phase was combined, washed with saturated brine (100 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was added with petroleum ether (100 mL), filtered, and the filter cake was dried to obtain compound M8-2. 1 H NMR (400 MHz, DMSO-d6) δ ppm 8.40 (d, J = 2.4 Hz, 1H), 7.65 (d, J = 2.4 Hz, 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 methyl magnesium chloride (3 M tetrahydrofuran solution, 23.7 mL) was added under nitrogen protection at 0 °C. The reaction solution was stirred at 0 °C for 2 hours. The reaction solution was poured into saturated ammonium chloride (100 mL) to quench, and the solution was extracted with ethyl acetate (100 mL*3). The organic phase was combined, washed with saturated brine (100 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was purified by a flash chromatographic column (silica gel, eluent methanol / dichloromethane, methanol ratio: 0~10%) to obtain compound M8-3. LCMS: m / z = 298.1, 300.1 [M+1] + . 1HNMR (400 MHz, DMSO-d6) δ ppm 8.38 (d, J = 2.4 Hz, 1 H), 7.53 (d, J = 2.4 Hz, 1 H), 3.39 - 3.47 (m, 4 H), 2.54 (s, 3 H), 2.40 - 2.45 (m, 4 H), 2.22 (s, 3 H).

[0414] Step 3

[0415] To formic acid (5.27 g, 109.67 mmol) was added triethylamine (57.1 g, 563.9 mmol) dropwise at 0 °C under nitrogen protection, then (S,S)-N-(p-toluenesulfonyl)-1,2-diphenylethanedi-amine (p-cymene) chlororuthenate (140 mg, 220 μmol) was added. The mixture was stirred at 40 °C for 15 min. Then it was cooled to room temperature, and compound M8-3 (6.54 g, 21.93 mmol) was added in portions. The reaction was warmed to 50 °C and stirred for 12 h. The reaction was directly concentrated, and the crude product was purified by flash column (silica gel, methanol / methylene chloride, methanol ratio: 0-10%) to give compound M8-4. LCMS: m / z = 300.0, 302.0 [M+1] + .

[0416] Step 4

[0417] Compound M8-4 (1 g, 3.33 mmol) was dissolved in N,N-dimethylformamide (10 mL) under nitrogen protection, and cooled to 0 °C, then sodium hydride (160 mg, 4.00 mmol, purity 60%) was added in portions. The solution was stirred at 0 °C for 1 h, then iodomethane (520 mg, 3.68 mmol) was added dropwise, and the reaction was continued at 0 °C for 2 h. The reaction was quenched by adding saturated ammonium chloride (50 mL) dropwise, and the solution was extracted with ethyl acetate (50 mL*3). The organic phase was combined and washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash column (silica gel, methanol / methylene chloride, methanol ratio: 0-10%) to give 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 bis(pinacolato)diboron (777 mg, 3.5 mmol) were dissolved in toluene (20 mL), potassium acetate (563 mg, 5.75 mmol) and 1,1-bis(diphenylphosphino)ferrocene palladium chloride (168 mg, 230 μmol) were added successively, replaced with nitrogen for three times, heated to 70 °C for 12 hours. The reaction solution was filtered, the filtrate was directly concentrated, and the obtained M8-6 crude product 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), water (20 mL), potassium carbonate (13.19 g, 95.46 mmol) was added, replaced with nitrogen for three times, 1,1-bis(diphenylphosphino)ferrocene palladium chloride (22.07 mg, 33.86 μmol) was added, and the reaction solution was stirred at 70 °C for 12 hours under nitrogen. 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 hydride (4 g, 100.00 mmol, purity 60%) was added in batches at 0 °C, and stirred for 30 min under nitrogen protection, then iodomethane (30.01 mmol, 2.4 mL) was added dropwise, the reaction solution was stirred at 0 °C for 1 hour, iodomethane (25.01 mmol, 2 mL) was added, and the reaction solution was stirred at 20 °C for 30 min. The reaction solution was quenched by adding 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 compound M8-8 crude product. 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. The reaction solution was stirred at 50 °C under nitrogen for 12 hours. The reaction solution was quenched with water (20 mL) and extracted with ethyl acetate (50 mL*3). The aqueous phase was extracted with dichloromethane (50 mL*3), and the organic phase was dried over anhydrous sodium sulfate and concentrated. The residue was purified by column chromatography (0-15% methanol / dichloromethane) to obtain a crude product. The crude product was stirred in acetonitrile (10 mL) for 10 minutes, filtered, and the filter cake was purified by prep-HPLC (column: C18 100*40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 13% to 43% of the proportion of acetonitrile in the mobile phase in 8 min) to obtain the trifluoroacetate salt of compound M8-9A (HPLC analysis method: column: ChromCore 120 C18 3 μm 3.0*30 mm; A phase: 4 liters of an aqueous solution containing 1.5 mL of trifluoroacetic acid, B phase: 4 liters of an acetonitrile solution containing 0.75 mL of trifluoroacetic acid; elution gradient: 10% to 80% of B phase in 6 minutes, 80% for 0.5 minutes, and 10% for 0.5 minutes; retention time of compound M8-9A: 3.205 min, and retention time of the isomer thereof: 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(pinacolato)diboron (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 at 90 °C under nitrogen 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 successively into 1.5 L of anhydrous dichloromethane, then 2-chloro-1-methylpyridinium iodide (266.10 g, 1.04 mol) was added portionwise into the above solution, and the mixture was stirred at 25 °C for 1 h. 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 (400 MHz, CDC13) 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 successively into 2 L of anhydrous tetrahydrofuran, and sodium borohydride (23.18 g, 612.58 mmol) was slowly added portionwise into the above solution at 0 °C. After the addition was completed, the mixture was continuously stirred at 0 °C for 2 h. After the reaction was completed, 500 mL of saturated aqueous ammonium chloride solution was slowly added dropwise, and the mixture was concentrated under reduced pressure to about 1 L of a residue, which was extracted with ethyl acetate (3*500 mL). The organic phase was washed with saturated sodium bicarbonate until the pH was about 8, the combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain compound M9-4. LCMS: m / z = 290.1 [M+1] + .

[0434] Compound M9-4 (220 g, 760.39 mmol) and N,N-diisopropylethylamine (147.41 g, 1.14 mol) were added successively into 2 L of anhydrous dichloromethane at 30 °C, then p-toluenesulfonyl chloride (159.46 g, 836.43 mmol) was added portionwise into the above solution, and the mixture was stirred at 25 °C for 3 h. 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 combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0~30%) to obtain compound M9-5. 1H NMR (400 MHz, CDC13) δ = 7.81 (br d, J = 8.0 Hz, 2H), 7.41-7.28 (m, 5H), 7.19 (br d, J = 7.2 Hz, 2H), 4.84-4.70 (m, 1H), 4.65 (br s, 1H), 4.29-4.18 (m, 2H), 3.26 (br d, J = 13.6 Hz, 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 into 900 mL of 1-methyl-2-pyrrolidinone successively, 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 extracted. The organic phase was washed with saturated brine (2*1 L) again, dried over anhydrous sodium sulfate, filtered, concentrated, and the residue was purified by flash column 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] Compound M9-6 (9.5 g, 23.80 mmol) was dissolved in 95 mL of anhydrous tetrahydrofuran at -78 °C, then lithium diisopropylamide 2M tetrahydrofuran n-heptane mixed solution (15.47 mL, 30.94 mmol) was added dropwise slowly to the mixture, and the mixture was stirred for half an hour under a nitrogen atmosphere. Di-tert-butyl azodicarboxylate (6.58 g, 28.56 mmol) was added in one portion to the above solution, and stirring was continued for half an hour. 1,3-Dimethyl-tetrahydro-2-pyrimidinone (91.50 g, 713.89 mmol) was slowly added to the above reaction solution, and the temperature was allowed to rise to room temperature naturally, and stirring was continued for 13 hours. After the reaction was completed, 100 mL of water was added for extraction, and lithium hydroxide monohydrate (3.00 g, 71.39 mmol) was added, and stirring was performed at room temperature for 1 hour. Concentration was performed, and the residue was diluted with 200 mL of saturated brine, and washed with ethyl acetate (3 * 200 mL), and the organic phase was discarded, and the aqueous phase was adjusted to pH 5 with 1N hydrochloric acid, and extracted with ethyl acetate (3 * 200 mL), and the organic phase was washed with saturated brine (2 * 200 mL), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by flash column 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] Compound M9-7 (1.6 g, 4.67 mmol) was dissolved in 32 mL of anhydrous methanol at 25 °C, and trimethylsilyldiazomethane 2M n-hexane solution (11.68 mL, 23.36 mmol) was added dropwise slowly to the mixture, and the mixture was stirred at 25 °C for 10 minutes. After the reaction was completed, 0.1 mL of acetic acid was added to quench the reaction. The reaction solution was concentrated to obtain compound M9-8. LCMS: m / z = 379.1 [M+23] + SFC analysis method detection (column: Cellulose-4 (100 mm * 4.6 mm, 3 μm; mobile phase: A phase supercritical carbon dioxide, B phase [0.05% diethylamine in isopropyl alcohol]; B%: 4 minutes from 5% to 40%, and 40% for 0.5 minutes, and then 5% for 1.5 minutes) The retention time of compound M9-8 was 1.342 min, and the chiral purity was 93.38%; the retention time of its enantiomer was 1.431 min, 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, the residue was added to 20 mL of methyl tert-butyl ether, 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 (400 MHz, D2O) δ = 4.30 (d, J = 3.8 Hz, 1H), 3.97 (q, J = 4.9 Hz, 1H), 3.73 (s, 3H), 2.93-2.83 (m, 1H), 2.54 (ddd, J = 4.5, 6.5, 11.5 Hz, 1H), 2.39 (td, J = 5.7, 11.7 Hz, 1H), 2.04 (dd, J = 9.4, 11.7 Hz, 1H), 1.76 (dd, J = 9.5, 12.0 Hz, 1H).

[0443] Reference Example 9: Compound M10

[0444]

[0445] Step 1

[0446] Compound M4-4 (9.2 g, 42.58 mmol), bis(pinacolato)diboron (16.22 g, 63.87 mmol), iridium (III) 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 for 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 150 mL of water and 150 mL of ethyl acetate were added for dilution, 400 mL of an aqueous base (7.5 g of sodium hydroxide and 30 g of sodium carbonate) was added to adjust the pH to 10, the liquid was separated, 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 combined organic phase was dried with 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), 1,10-phenanthroline (694 mg, 3.85 mmol) were added successively. The reaction was stirred at 60 °C for 2 hours. After the reaction was completed, the reaction was filtered, and the filtrate was directly concentrated. The crude product was purified by flash column 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), compound M10-3 (170 mg, 790.54 μmol, 2HCl) were dissolved in toluene (5 mL), and cooled to 0 °C, then cesium carbonate (1.29 g, 3.95 mmol), (R)-(+)-2,2-bis(diphenylphosphino)-1,1'-binaphthalene (50 mg, 80.95 μmol) and palladium acetate (36 mg, 158.91 μmol) were added. The reaction was replaced by nitrogen for 3 times, and stirred at 90 °C for 12 hours. After the reaction was completed, the reaction was directly concentrated, and the obtained crude product was purified by flash column 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), potassium carbonate (8.53 g, 61.75 mmol) were dissolved in methylpyrrolidone (25 mL), and tert-butyl cyanoacetate (4.65 g, 32.93 mmol) was added, and the reaction was stirred at 85 °C for 12 hours. After the reaction was completed, the reaction was cooled to room temperature, water (25 mL) was added, and the pH was adjusted to 2 with 3M hydrochloric acid. A solid was precipitated, and the solid was filtered and dried to obtain compound M11-2. 1 H NMR (400 MHz, CD3OD) δ ppm 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 (4 M, 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 hydroxide (1.48 g, 36.93 mmol, 60% purity) was added at 0°C. The mixture was stirred at 0°C for 20 minutes under nitrogen. 1,2-Dibromoethane (22.16 mmol, 1.67 mL) was then added and stirred at 25°C for 12 hours under nitrogen. After completion, the reaction was quenched with water (100 mL) and extracted with ethyl acetate (150 mL x 3). The organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the crude product. The crude product 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 completion of the reaction, 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), 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 reaction was stirred at 80 °C for 6 hours. After the reaction was completed, water (150 mL) was added to quench the reaction, and the reaction 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. Compound M11-6 was obtained by LCMS: m / z = 302.1, 304.1 [M+1] + . 1 H NMR (400 MHz, CD3OD) δ ppm 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 reaction 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), and 1,1-bis(diphenylphosphino)ferrocene palladium chloride (21 mg, 29.24 μmol) were dissolved in dioxane (5 mL) and water (1 mL), and the reaction was replaced with nitrogen three times. The reaction was stirred at 60 °C for 3 hours. After the reaction was completed, the reaction was directly concentrated, and the crude product was purified by flash column 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] Trimethylsulfoxonium iodide (139 mg, 629.24 μmol), potassium tert-butoxide (71 mg, 629.24 μmol) were added into dimethyl sulfoxide (3 mL) and stirred at 50 °C for 2 hours. Then the solution of compound M12-2 (0.05 g, 125.85 μmol) in dimethyl sulfoxide (1 mL) was added dropwise at 25 °C, and the reaction was stirred at 80 °C for 12 hours. After the reaction was completed, the reaction was diluted with water (50 mL) and extracted with ethyl acetate (50 mL*3). The organic phase was washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-50%) to give compound M12. LCMS: m / z = 410.9, 412.9 [M+1] + .

[0471] Example 1

[0472]

[0473]

[0474] Step 1

[0475] To a mixture 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)ferrocene palladium chloride (794.48 mg, 1.09 mmol) were added. The mixture was stirred at 85 °C for 4 hours under nitrogen. The reaction was concentrated. The residue was diluted with 100 mL of water and 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 column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to give compound 1-1. LCMS: m / z = 655.1, 657.1 [M+1] + .

[0476] Step 2

[0477] To a solution of compound 1-1 (4.5 g, 6.86 mmol) in DMF (50 mL) at 0°C, iodoethane (2.14 g, 13.73 mmol) and cesium carbonate (4.47 g, 13.73 mmol) were added sequentially. 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 x 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 a tetrahydrofuran solution of tetrabutylammonium fluoride (1 M, 34.81 mL) were added sequentially 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 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. 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), bis(pinacolato)diboron (406 mg, 1.80 mmol) were dissolved in toluene (10 mL), 1,1'-bis(diphenylphosphino)ferrocene palladium chloride (131 mg, 179.62 μmol) and potassium acetate (264 mg, 2.69 mmol) were added successively, the reaction solution was replaced with nitrogen for 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 boronic 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'-bis(t-butylphosphino)ferrocene palladium 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 the reaction solution was replaced with nitrogen for 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 column 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 about 5, and the solution was extracted with ethyl acetate (50 mL*3). The organic phase was 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), then N,N-diisopropylethylamine (260 mg, 2.01 mmol, 0.35 mL), 1-hydroxybenzotriazole (0.044 g, 325.63 μmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.35 g, 1.83 mmol) were added. The reaction was stirred at 20 °C for 12 h. The reaction was diluted with water (50 mL), and the solution was extracted with dichloromethane (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0~80%) to give compound 1-8A. LCMS: m / z = 743.4 [M+1] + .

[0488] Step 8

[0489] Compound 1-8A (20.00 mg, 26.92 μmol) was dissolved in dichloromethane (1 mL), then trifluoroacetic acid (1.54 g, 13.46 mmol, 1 mL) was added. The reaction was stirred at 15 °C for 12 h. The reaction was concentrated to give a crude product of trifluoroacetate 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), then N,N-diisopropylethylamine (311.13 μmol, 54 μL), (1S,2S)-2-methylcyclopropane-1-carboxylic acid (7 mg, 62.3 μmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (36 mg, 93.4 μmol) were added. The reaction was stirred at 20 °C for 1.5 h. The reaction was diluted with water (20 mL), and the solution was extracted with ethyl acetate (20 mL*3). The organic phases were combined, washed with saturated brine (20 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give a crude product, which was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0~80%) to give compound 1A. LCMS: m / z = 725.3 [M+1] + ; 1H NMR (400 MHz, CD3OD) δ ppm 8.64 (dd, J = 4.8, 1.6 Hz, 1 H), 8.12 (s, 1 H), 7.79 (dd, J = 7.6, 1.6 Hz, 1 H), 7.54 (d, J = 8.4 Hz, 1 H), 7.44 (dd, J = 7.6, 4.8 Hz, 1 H), 7.34 - 7.39 (m, 2 H), 6.02 (t, J = 6.2 Hz, 1 H), 4.03 - 4.13 (m, 2 H), 3.71 - 3.86 (m, 3 H), 3.43 - 3.56 (m, 2 H), 3.03 (s, 3 H), 2.64 - 2.72 (m, 1 H), 2.37 - 2.44 (m, 1 H), 2.06 - 2.13 (m, 1 H), 1.89 - 1.97 (m, 2 H), 1.70 - 1.80 (m, 1 H), 1.48 - 1.60 (m, 3 H), 1.32 - 1.37 (m, 2 H), 1.18 - 1.23 (m, 6 H), 0.90 - 0.97 (m, 2 H), 0.76 - 0.83 (m, 2 H), 0.57 - 0.66 (m, 6 H), 0.45 - 0.51 (m, 1 H).

[0492] Example 2

[0493]

[0494] Step 1

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

[0496] Step 2

[0497] Compound 2-1A (300 mg, 609.19 pmol) and compound M3 (492.89 mg, 913.79 pmol) were dissolved in a mixed solvent of 1,4-dioxane (3 mL), toluene (3 mL), water (0.5 mL), potassium phosphate (387.94 mg, 1.83 mmol) was added, and 1,1-bis(tert-butylphosphino)ferrocene palladium chloride (39.70 mg, 60.92 pmol) was added. 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 to obtain a crude product. The crude product 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 pmol) was dissolved in a mixed solution of tetrahydrofuran (3 mL), 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, then the reaction solution was adjusted to neutral pH with 1M dilute hydrochloric acid, and then extracted with ethyl acetate (5 mL x 3). The combined organic phase was 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 pmol) was dissolved in dichloromethane (30 mL), and 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. The reaction solution was stirred at 25 °C for 12 hours. The reaction solution was concentrated under reduced pressure to obtain a crude product. The crude product was purified by silica gel column chromatography (0-30% ethyl acetate / petroleum ether), and then prepared and separated by high performance liquid chromatography (column: C18 100 x 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 was stirred at 25 °C for 5 h, 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 salt), (1S,2S)-2-methylcyclopropane-1- carboxylic acid (3.76 mg, 37.57 μmol) was dissolved in N,N-dimethylformamide (1 mL), then N,N-diisopropylethylamine (9.71 mg, 75.13 μmol, 13.09 μL) was added under stirring, O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (19.05 mg, 50.09 μmol) was added, the reaction was stirred at 25 °C for 12 h, extracted with ethyl acetate (5 mL x 3), the organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, the residue was concentrated under reduced pressure, and purified by high performance liquid chromatography (column: C18 100 x 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 (CDC13, 400 MHz) δ 9.1-9.2 (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 = 5.8 Hz, 1H), 4.11 (br d, J = 10.5 Hz, 1H), 4.0-4.1 (m, 1H), 3.88 (br d, J = 10.3 Hz, 1H), 3.64 (br d, J = 15.6 Hz, 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 = 5.8 Hz), 1.2-1.4 (m, 8H), 1.1-1.2 (m, 1H), 1.07 (br d, 3H, J = 5.5 Hz), 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(pinacolato)diboron (52.89 g, 208.26 mmol), iridium (III) 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 successively into 600 mL of anhydrous tetrahydrofuran, which was replaced by nitrogen for 3 times. 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, the pH was adjusted to 10 by adding basic water (400 mL of an aqueous solution of 10 g of NaOH and 40 g of sodium carbonate), and the mixture was separated, the organic phase was discarded, the aqueous phase was adjusted to pH 6 with concentrated hydrochloric acid, and extracted with 500 mL of ethyl acetate for 3 times, the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain compound 3-1. 1 H NMR (400 MHz, CDCl3) δ = 8.93 (d, J = 1.6 Hz, 1H), 8.22 (d, J = 1.6 Hz, 1H), 4.96 (q, J = 6.4 Hz, 1H), 3.32 (s, 3H), 1.50 (d, J = 6.4 Hz, 3H), 1.37 (s, 12H).

[0511] Step 2

[0512] Compound 3-1 (40 g, 116.95 mmol) was slowly added dropwise into a 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) in 1.5 L of acetonitrile at 80 °C. 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 column 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 found 434.3, 436.3; 1H NMR (400 MHz, CDC13) δ = 8.23 (d, J = 2.4 Hz, 1H), 7.39-7.25 (m, 5H), 7.23 (d, J = 2.4 Hz, 1H), 5.09 (s, 2H), 4.78 (q, J = 6.4 Hz, 1H), 3.68-3.55 (m, 4H), 3.21 (s, 3H), 3.13 (br s, 4H), 1.39 (d, J = 6.4 Hz, 3H).

[0513] Step 3

[0514] Compound 3-2 (7 g, 16.12 mmol), bis(pinacolato)diboron (5.46 g, 24.18 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium chloride (589.65 mg, 805.85 μmol) and potassium acetate (3.95 g, 40.29 mmol) were added into 140 mL of anhydrous dioxane, the mixture was stirred at 80 °C for 20 hours under nitrogen. The reaction was concentrated. The residue was purified by flash column chromatography (silica gel, methanol / methylene chloride, methanol ratio: 0-10%) to give 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)ferrocene palladium chloride (824.72 mg, 1.13 mmol) and potassium phosphate (11.96 g, 56.36 mmol) were added into 250 mL of anhydrous dioxane and 80 mL of water, the mixture was stirred at 70 °C for 12 hours under nitrogen. The reaction 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, the filtrate was concentrated under reduced pressure, the residue was purified by flash column chromatography (silica gel, ethyl acetate / petroleum ether, ethyl acetate ratio: 0-80%) to give 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 sequentially to 100 mL of anhydrous DMF, 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 and extracted with ethyl acetate (3 x 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 in tetrahydrofuran (1 M, 13.75 mL) were added sequentially 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(chloronaphthalene) boronate (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 sequentially to 50 mL of anhydrous dioxane. The atmosphere was purged 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-bis(tert-butylphosphino)ferrocene palladium chloride (34.85 mg, 53.47 μmol) and potassium phosphate (283.74 mg, 1.34 mmol) were added into 15 mL of dioxane and 5 mL of water, respectively, and the mixture was stirred at 70 °C for 4 h under nitrogen. The reaction was concentrated. The residue was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to give 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 into 20 mL of tetrahydrofuran and 20 mL of water, respectively, and the mixture was stirred at 15 °C for 20 h. The reaction was concentrated. The residue was diluted with 5 mL of water and adjusted to pH 6-7 by dropwise addition of 1 N dilute hydrochloric acid, extracted with ethyl acetate (3*30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 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 into 65 mL of acetonitrile, respectively, and the mixture was stirred at 25 °C for 40 h. The reaction was concentrated. The residue was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0-100%) to give compound 3-10A. LCMS: m / z = 961.4 [M+1] + .

[0529] Step 11

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

[0531] Step 12

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

[0533] Step 13

[0534] Compound 3-12A (45 mg, trifluoroacetate salt), compound (1S,2S)-2-methylcyclopropane-1- carboxylic acid (9.12 mg, 91.10 pmol), N,N-diisopropylethylamine (303.66 pmol, 52.89 pL) and HATU (69.28 mg, 182.20 pmol) were added to 5 mL of anhydrous DMF sequentially, 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 (preparative method: column type: C18 100 x 40 mm; mobile phase: [water (trifluoroacetic acid) - acetonitrile]; gradient (acetonitrile %): from 14% to 44% in 8 minutes) to give the trifluoroacetate salt of compound 3A. LCMS: m / z = 823.4 [M+1] + ; 1H NMR (400 MHz, CDC13) δ = 8.67 (br s, 1H), 8.38 (s, 1H), 7.57 (br d, J = 8.4 Hz, 1H), 7.37 (d, J = 8.4 Hz, 1H), 7.27 (s, 1H), 7.20 (s, 1H), 6.78 (br d, J = 7.6 Hz, 1H), 5.91 (br s, 1H), 4.22 (br d, J = 6.0 Hz, 1H), 4.14-4.06 (m, 2H), 3.99-3.91 (m, 2H), 3.67 (br s, 4H), 3.53 (br dd, J = 5.6, 14.8 Hz, 4H), 3.27 (s, 3H), 3.26-2.99 (m, 3H), 2.92 (br s, 3H), 2.81 (br d, J = 14.4 Hz, 1H), 2.52 (br d, J = 14.2 Hz, 1H), 2.12-1.71 (m, 4H), 1.47 (br d, J = 6.0 Hz, 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 pmol), (1r,2R,3S)-2,3-dimethylcyclopropyl-1- carboxylic acid (31 mg, 269.98 pmol) were dissolved in N,N-dimethylformamide (2 mL), then N,N-diisopropylethylamine (175 mg, 1.35 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (154 mg, 404.9 pmol) were added. The reaction was stirred at 20 °C for 2 hours. The reaction was filtered, and the filtrate was directly purified by HPLC prep (purification method: column type: C18 100 x 40 mm; mobile phase: [water (trifluoroacetic acid) - acetonitrile]; gradient (acetonitrile %): 16% up to 46% in 8 minutes) to give trifluoroacetate salt of compound 4A. LCMS: m / z = 837.4 [M+1] + ; 1H NMR (400 MHz, CD3OD) δ ppm 8.53 (s, 1 H), 8.27 (s, 1 H), 7.93 (s, 1 H), 7.70 (d, J = 8.8 Hz, 1 H), 7.49-7.57 (m, 2 H), 6.16 (t, J = 6.0 Hz, 1 H), 4.95-5.04 (m, 3 H), 4.10-4.28 (m, 3 H), 3.88-4.06 (m, 4 H), 3.45-3.70 (m, 3 H), 3.24 (s, 3 H), 3.02 (s, 3 H), 2.82-2.90 (m, 1 H), 2.46-2.55 (m, 1 H), 2.01-2.11 (m, 1 H), 1.81-1.91 (m, 1 H), 1.68-1.74 (m, 1 H), 1.58-1.66 (m, 1 H), 1.42-14.7 (m, 3 H), 1.26-1.41 (m, 7 H), 1.17-1.23 (m, 3 H), 1.08-1.15 (m, 7 H), 0.75-0.85 (m, 6 H).

[0538] Example 5

[0539]

[0540] Step 1

[0541] Compound M8 (200 mg, 338.64 μmol), compound M3 (274 mg, 507.98 μmol) were dissolved in dioxane (4 mL), toluene (1.3 mL), water (1.3 mL), potassium phosphate (215.65 mg, 1.02 mmol), 1,1-bis(tert-butylphosphine)ferrocenepalladium chloride (22.07 mg, 33.86 μmol) were added, the reaction was stirred at 70 °C for 12 hours under nitrogen atmosphere. The reaction was filtered, the filtrate was concentrated under reduced pressure to obtain a crude product, the crude product 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 mixture solution of tetrahydrofuran (2 mL), water (0.5 mL), lithium hydroxide monohydrate (46.59 mg, 1.11 mmol) was added, the reaction was stirred at 25 °C for 1 hour, then the reaction was adjusted to pH 7-8 with 1M dilute hydrochloric acid, then extracted with ethyl acetate (10 mL*5), the organic phase was washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, 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), and then tetramethylchloroformamidinium hexafluorophosphate (21.16 mg, 75.41 μmol), N-methylimidazole (12.38 mg, 150.83 μmol) were added. The reaction solution was stirred at 25 °C for 1 h. The reaction solution was added with water (20 mL), and extracted with dichloromethane (5 mL*3). The organic phase was washed with saturated brine (5 mL), dried over anhydrous sodium sulfate, and concentrated to give 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), and then trifluoroacetic acid (767.50 mg, 6.73 mmol, 0.5 mL) was added. The reaction solution was stirred at 25 °C for 1 h. The reaction solution was concentrated under reduced pressure to give the crude trifluoroacetate salt of compound 5-4A, which was directly used in the next step.

[0548] Step 5

[0549] The crude trifluoroacetate salt of compound 5-4A obtained in Step 4 was dissolved in N,N-dimethylformamide (1 mL), and then (1r,2R,3S)-2,3-dimethylcyclopropyl-1-carboxylic acid (9.70 mg, 84.95 μmol) was added, followed by the addition of N,N-diisopropylethylamine (21.96 mg, 169.90 μmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (43.07 mg, 113.27 μmol) under stirring. The reaction solution was stirred at 25 °C for 3 h, and then purified by prep-HPLC (column: C18 100*40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: 8 min from 16% to 46% acetonitrile) to give the trifluoroacetate salt of compound 5A. LCMS: m / z = 873.9 [M+1] +1H NMR (CD3OD, 400 MHz) δ 8.5-8.6 (m, 1H), 8.2-8.3 (m, 1H), 7.7-7.7 (m, 1H), 7.61 (br d, J = 2.3 Hz, 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] dichloropalladium (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 combined organic phase was 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), N,N-diisopropyl ethylamine (952.45 μmol, 165.90 μL) and 2-(7-azobenzo-triazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (43.46 mg, 114.29 μmol) were added, and the mixture was stirred at 25 °C for half an hour. After the reaction was completed, saturated brine (30 mL) was added, and the mixture was extracted with ethyl acetate (30 mL) and tetrahydrofuran (30 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 6-3A which was used directly 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. The mixture was stirred at 25 °C for half an hour. The pH value of the system was adjusted to 7.0 with 1 M hydrochloric acid, and then diluted with saturated brine (50 mL). The mixture was extracted twice with ethyl acetate (30 mL) and tetrahydrofuran (30 mL), and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give compound 6-4A which was used directly 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'-tetramethylchloroformamidinium hexafluorophosphate (597.68 mg, 2.13 mmol) and N-methylimidazole (174.89 mg, 2.13 mmol, 169.80 μL) were added. The mixture was stirred at 25 °C for 1 hour under nitrogen protection. The reaction solution was concentrated, and then separated by pre-TLC preparation (DCM / MeOH = 10:1) to give compound 6-5A. LCMS: m / z = 842.0 [M+1] + .

[0563] Step 6

[0564] Compound 6-5A (52 mg, 61.83 pmol) was dissolved in dichloromethane (2 mL), trifluoroacetic acid (798.20 mg, 7.00 mmol, 520.00 pL) was added, and the mixture was stirred at 25 °C for 1 h. The reaction solution was concentrated to give 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 pmol) and (1r,2R,3S)-2,3-dimethylcyclopropyl-1- carboxylic acid (9.35 mg, 81.8 pmol) were dissolved in DMF (2 mL), 2-(7-azobenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (31.13 mg, 81.87 pmol) and N,N-diisopropylethylamine (409.37 pmol, 71.30 pL) were added, and the mixture was stirred at 25 °C for 1 h. The reaction solution was diluted with saturated brine (50 mL), extracted with ethyl acetate (30 mL) and tetrahydrofuran (30 mL) twice, and the combined organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The crude product was purified by pre-TLC (developing agent: dichloromethane / methanol = 10:1), and then by SFC separation and purification (column: DAICEL CHIRALCEL OD (250 mm*30 mm, 10 pm); mobile phase: [A phase: supercritical carbon dioxide, B phase: ethanol (0.1% ammonia water)]; gradient (B%): 50%, isocratic elution) to give compounds 6A and 6B.

[0567] SFC analysis (column: Chiralcel OD-3 50*4.6 mm I.D., 3 pm, mobile phase: A: supercritical carbon dioxide B: ethanol (containing 0.05% diethylamine), gradient elution: mobile phase B from 5% to 40% in two minutes, then 40% for 1.2 minutes, and then 5% for 0.8 minutes) showed that compound 6A had an RT of 1.896 min and an ee of 96.98%, and compound 6B had an RT of 2.201 min and an ee of 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+l] + ; 1 H 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 pmol) and (1S,2S)-2-methylcyclopropyl-1- carboxylic acid (14.05 mg, 81.8 pmol) were dissolved in DMF (10 mL), 2-(7- azobenzotriazol)-N,N,N',N'-tetramethyluronium hexafluorophosphate (53.37 mg, 140.35 pmol) and N,N-diisopropylethylamine (72.56 mg, 561.42 pmol) were added, stirred at 25 °C for 1 h. Diluted with saturated brine (50 mL), extracted with ethyl acetate (30 mL) and tetrahydrofuran (30 mL) for 2 times, the combined organic phase was dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, the obtained crude was purified by flash silica gel column (mobile phase: dichloromethane / methanol = 100:1), then purified by SFC separation (column: DAICEL CHIRALCEL OD (250 mm*30 mm, 10 pm); mobile phase: [A phase is supercritical carbon dioxide, B phase is ethanol (0.1% ammonia water)]; gradient (B%): 50%) to obtain compounds 7A and 7B. SFC analysis (method: column: Chiral OD-3 100*4.6 mm I.D., 3 pm, mobile phase: A: supercritical carbon dioxide, B: ethanol (containing 0.05% diethylamine), gradient (B%): 40%), the RT of compound 7A was 1.365 min, ee = 100%; the RT of compound 7B was 2.325 min, ee = 88.2%. Compound 7A: LCMS: m / z = 823.7 [M+1] + ; 1H 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+l] + ; 1H NMR (400 MHz, CD3OD) δ = 8.38 (s, 1H), 8.33 (d, J = 2.8 Hz, 1H), 7.60-7.54 (m, 1H), 7.39-7.34 (m, 2H), 7.31 (d, J = 3.0 Hz, 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 = 14.3 Hz, 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 = 9.5 Hz, 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 = 6.0 Hz, 2H), 0.91 (td, J = 4.3, 8.5 Hz, 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 starting materials, the reaction mixture was directly purified by high performance liquid chromatography (column: C18 100 x 40 mm; mobile phase: [water (trifluoroacetic acid) - acetonitrile]; gradient: acetonitrile from 16% to 46% in 8 min) according to the synthetic method of Example 7 to give the trifluoroacetate salt of compound 8A. LCMS: m / z = 859.5 [M+1] + . 1H NMR (400 MHz, DMSO-d6) d = 8.88 (br d, J = 9.0 Hz, 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 = 11.3 Hz, 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 = 1.5 Hz, 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 = 6.3 Hz, 4H), 1.05 (br t, J = 6.9 Hz, 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), then tetrabutylammonium fluoride (1M tetrahydrofuran solution, 65.60 mL) was added. The reaction was stirred at 50 °C for 16 hours. After the reaction was completed, the reaction 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 phase was combined and washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash column 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), then triethylamine (3.56 g, 35.19 mmol), 4-dimethylaminopyridine (72 mg, 596.51 μmol) were added, and the solution was cooled 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 min. 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), and the organic phase was combined and washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash column 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(pinacolato)diboron (6.66 g, 26.22 mmol), potassium acetate (2.57 g, 26.22 mmol), 1,1-bis(diphenylphosphino)ferrocene palladium chloride (0.768 g, 1.05 mmol) were dissolved in toluene (40 mL), and the solution was replaced with nitrogen for 3 times, and the reaction solution was stirred at 90 °C for 3 h. After the reaction was completed, the reaction solution was directly concentrated. The crude product was purified by flash column 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)ferrocene palladium chloride (750 mg, 1.12 mmol) were dissolved in toluene (30 mL), dioxane (10 mL) and water (10 mL), and the solution was replaced with nitrogen for 3 times, and the reaction solution was stirred at 70 °C for 12 h. After the reaction was completed, the reaction solution was directly concentrated. The crude product was purified by flash column 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, then sodium bicarbonate (1.03 g, 12.28 mmol) and silver trifluoromethanesulfonate (3.15 g, 12.25 mmol) were added, then iodine (2.33 g, 9.18 mmol) in tetrahydrofuran (5 mL) was added dropwise. The reaction was stirred at 0 °C for 15 min. After the reaction was completed, the reaction was quenched by adding saturated sodium sulfite solution (100 mL) at 0 °C, the resulting solution was extracted with ethyl acetate (100 mL*3), the organic phase was combined and washed with saturated brine (100 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was purified by flash column 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 was stirred at 25 °C for 12 h. After the reaction was completed, the reaction was adjusted to pH about 6 by adding saturated citric acid solution, the resulting solution was extracted with ethyl acetate (100 mL*3), the organic phase was combined and washed with saturated brine (100 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was purified by high performance liquid chromatography preparation (column: C18 100*40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile from 30% to 60% in 8 min) 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), then N,N-diisopropylethylamine (1.44 g, 11.18 mmol), compound M9 (0.67 g, 1.12 mmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (510 mg, 1.44 mmol) were added. The reaction was stirred at 25 °C for 2 hours. After the reaction was completed, the reaction was diluted with water (50 mL), and the solution was extracted with ethyl acetate (50 mL*3), the organic phases were combined and washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was purified by flash column 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] Compound 9-7A (0.63 g, 854.07 µmol) was dissolved in tetrahydrofuran (6 mL) and methanol (6 mL), 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 was stirred at 0 °C for 1 hour. After the reaction was completed, the reaction was adjusted to pH about 6 with saturated citric acid solution, and the solution was extracted with ethyl acetate (50 mL*3), the organic phases were combined and washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was purified by flash column 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), then N-methylimidazole (1.13 g, 13.82 mmol,) and N,N,N,N-tetramethylchloroformamidinium hexafluorophosphate (388 mg, 1.48 mmol) were added. The reaction was stirred at 80 °C for 2 hours. After the reaction was completed, the reaction was directly concentrated to remove acetonitrile, then diluted with water (30 mL), and the solution was extracted with ethyl acetate (30 mL*3), the organic phases were combined and washed with saturated brine (30 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was purified by flash column 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-bis(cyclohexylphosphino)-2,6-dimethoxybiphenyl (30 mg, 71.86 μmol), tris(dibenzylideneacetone)dipalladium (26 mg, 28.54 μmol) were dissolved in toluene (5 mL), and pinacolborane (145.10 mg, 1.13 mmol) was added dropwise at 0 °C under nitrogen. The reaction was stirred at 60 °C for 3 h under nitrogen. After the reaction was completed, the reaction was quenched by dropwise addition of saturated ammonium chloride (10 mL), and the solution was extracted with ethyl acetate (50 mL*3). The organic phase was washed once with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The obtained crude product was purified by flash column 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)ferrocene palladium chloride (21 mg, 28.54 μmol) were dissolved in toluene (3 mL), dioxane (1 mL) and water (1 mL), and the reaction was stirred at 65 °C for 12 h after nitrogen replacement for 3 times. After the reaction was completed, the reaction was directly concentrated, and the crude product 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 was stirred at 25 °C for 12 h. After the reaction was completed, the reaction was filtered, and the obtained filtrate was separated and purified by high performance liquid chromatography preparation (column: C18 100*40 mm; mobile phase: [water (trifluoroacetic acid)-acetonitrile]; gradient: acetonitrile from 22% to 52% in 8 min) to obtain trifluoroacetate 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 (2 M, 2 mL), the reaction was stirred at 25 °C for 1 h. After the reaction was completed, the reaction was directly concentrated, and the crude 9-13A was directly used for the next step. LCMS: m / z = 783.3 [M+1] + .

[0607] Step 14

[0608] Compound 9-13A (0.03 g, 38.31 μmol), (1S,2S)-2-methylcyclopropane-1-carboxylic acid (8 mg, 79.91 μmol) was dissolved in N,N-dimethylformamide (2 mL), then N,N-diisopropylethylamine (50 mg, 386.87 μmol) and O-(7-azabenzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (30 mg, 78.90 μmol) were added. The reaction was stirred at 25 °C for 12 h. After the reaction was completed, the reaction was filtered, and the filtrate was separated and purified by high performance liquid chromatography preparation (column: C18 100 x 40 mm; mobile phase: [water (trifluoroacetic acid) - acetonitrile]; gradient: acetonitrile from 17% to 47% in 8 min) to give trifluoroacetate salt of compound 9A. LCMS: m / z = 865.3 [M+1] + . 1 HNMR (400 MHz, CD3OD) δ ppm 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), 2.09-1.95 (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), cesium carbonate (2.61 g, 8.01 mmol) were dissolved in 1-methyl-2-pyrrolidinone (10 mL), and the solution was cooled to 0 °C, followed by dropwise addition of trifluoroethyl triflate (2.66 g, 11.44 mmol). The reaction solution was stirred at 25 °C for 48 h. 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 phase was combined and washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash column 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] To compound 10-1A (0.5 g, 523.01 µmol) was added tetrabutylammonium fluoride (1M tetrahydrofuran solution, 5 mL) at 0 °C. The reaction solution was stirred at 40 °C for 12 h. 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 phase was combined and washed with saturated brine (50 mL) once, dried over anhydrous sodium sulfate, filtered, and the filtrate was directly concentrated. The crude product was purified by flash column chromatography (silica gel, eluent ethyl acetate / petroleum ether, ethyl acetate ratio: 0~80%) to obtain compound 10-2A (LC-MS analysis method: 5-95AB_1.5min, the retention time of compound 10-2A was 1.009 min, and the retention time of its isomer was 0.981 min). LCMS: m / z = 717.1 [M+1] + .

[0616] Steps 3~11

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

[0618] Example 11

[0619]

[0620] The reaction solution of compound 11A was obtained according to the synthetic procedure of Reference Example 6-9. The reaction solution was diluted with water until no more solid precipitated, filtered, and the solid was collected. The solid was then purified by preparative HPLC (column: C18 100 x 40 mm; mobile phase: [water (trifluoroacetic acid) - acetonitrile]; gradient: acetonitrile from 15% to 45% over 8 minutes) to give the trifluoroacetate salt of compound 11A. LCMS: m / z = 825.4 [M+1] +1H NMR (400 MHz, CDC13) 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, 12.8 Hz, 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 = 5.5 Hz, 3H), 1.44-1.36 (m, 1H), 1.31-1.19 (m, 3H), 1.13 (br d, J = 5.8 Hz, 3H), 1.02-0.93 (m, 6H), 0.73-0.64 (m, 1H), 0.49-0.34 (m, 3H).

[0621] Example 12

[0622]

[0623] The reaction solution of compound 12A was obtained according to the synthetic procedure of Reference Example 6-9. The reaction solution was added water until no more solid precipitated, suction filtered, and the solid was collected, then purified by preparative high performance liquid chromatography (column: C18 100 x 40 mm; mobile phase: [water (trifluoroacetic acid) - acetonitrile]; gradient: acetonitrile from 17% to 47% in 8 minutes) to give the trifluoroacetate salt of the target compound 12A. LCMS: m / z = 862.2 [M+23] + . 1H NMR (400 MHz, CDC13) 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 (br d, 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), sodium bicarbonate (1.44 g, 17.14 mmol) and Boc20 (1.17 g, 5.34 mmol) were added, stirred at 25 °C for 12 h. After the reaction was completed, the reaction mixture was extracted with water (50 mL) and ethyl acetate (50 mL*3). The organic phase was dried and concentrated, and then purified by SFC (column: DAICEL CHIRALPAK IG (250 mm*30 mm, 10 um); mobile phase: [A phase: carbon dioxide supercritical fluid, B phase: isopropyl alcohol containing 0.1% ammonia water]; B phase gradient elution ratio 30% etc.) to obtain compound 13-1A (Rt = 3.625; SFC analysis method: column: DAICEL CHIRALPAK IG (100*4.6 mm I.D., 3 um); mobile phase: [A phase: carbon dioxide supercritical fluid, B phase: isopropyl alcohol containing 0.05% ammonia water]; B phase from 5% to 40% in 4.5 min, then 5% for 1.5 min, flow rate 2.5 mL / min, column temperature: 40 °C) and 13-1B (Rt = 4.200; SFC analysis method: column: DAICEL CHIRALPAK IG (100*4.6 mm I.D., 3 um); mobile phase: [A phase: carbon dioxide supercritical fluid, B phase: isopropyl alcohol containing 0.05% ammonia water]; B phase from 5% to 40% in 4.5 min, then 5% for 1.5 min, flow rate 2.5 mL / 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(pinacolato)diboron (688.76 mg, 2.71 mmol) were added to toluene (10 mL), and potassium acetate (354.91 mg, 3.62 mmol) was added, and the reaction was replaced with nitrogen, and 1,1-bis(diphenylphosphino)ferrocene palladium chloride (132.31 mg, 180.82 μmol) was added, and the reaction was stirred at 70 °C under nitrogen for 12 h. The reaction 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), 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, stirred at 0 °C for 0.5 h, then trifluoroacetate of compound M9 (35.81 mg, 132.54 μmol) was added at 0 °C, stirred at 25 °C for 1 h, after the reaction was completed, extracted with water (50 mL) and ethyl acetate (50 mL*3), 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), compound 13-2A (89.63 mg, 126.11 μmol) were dissolved in dioxane (1 mL), water (1 mL) and toluene (3 mL), then potassium phosphate (64.25 mg, 302.67 μmol) and 1,1-bis(tert-butylphosphine)ferrocene palladium chloride (13.15 mg, 20.18 μmol) were added, stirred at 70 °C for 12 h under nitrogen protection, after the reaction was completed, water (10 mL) was added, then extracted with ethyl acetate (10 mL*3), 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] The reaction solution of compound 13A was obtained according to the synthetic method of Reference Examples 6-9. Water (10 mL) was added to the reaction solution, extracted with ethyl acetate (10 mL*3), the organic phase was dried and concentrated, the crude product was separated by thin layer chromatography preparation plate (dichloromethane:methanol = 10:1) to obtain compound 13A. LCMS: m / z = 850.0 [M+1] + . 1H NMR (400 MHz, CD3OD) δ ppm 0.56 (br s, 3 H) 0.66 (br s, 1 H) 0.90 (br s, 3 H) 1.01 (br s, 3 H) 1.06 (br t, J=7.15 Hz, 6 H) 1.48 (br s, 2 H) 1.58 (br d, J=9.29 Hz, 1 H) 1.94 (br s, 2 H) 2.10 (br s, 1 H) 2.19 - 2.28 (m, 1 H) 2.40 (br d, J=5.02 Hz, 2 H) 2.59 - 2.64 (m, 1 H) 2.73 - 2.77 (m, 3 H) 2.80 (s, 1 H) 2.95 (d, J=7.28 Hz, 1 H) 3.03 (s, 3 H) 3.09 - 3.17 (m, 8 H) 3.43 (br d, J=9.03 Hz, 3 H) 3.58 - 3.67 (m, 7 H) 3.87 (s, 1 H) 7.36 (s, 1 H) 7.38 - 7.42 (m, 2 H) 7.59 (d, J=8.78 Hz, 1 H) 8.34 - 8.38 (m, 2 H).

[0636] Example 14

[0637]

[0638] Compound 14A was prepared from compound 13-1B according to the synthetic method of Example 13.

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

[0640] (s, 3H) 1.04 (br s, 3H) 1.13 (br d, J = 5.77 Hz, 4H) 1.24 - 1.29 (m, 1H) 1.31 (br s, 2H) 1.33 - 1.38 (m, 2H) 1.44 (br d, J = 6.02 Hz, 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 = 10.04 Hz, 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 = 10.79 Hz, 1H) 3.74 (br d, J = 12.55 Hz, 1H) 4.24 (br d, J = 6.53 Hz, 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 = 8.53 Hz, 1H) 8.43 (s, 2H).

[0641] Example 15

[0642]

[0643] Step 1

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

[0645] Step 2

[0646] Compound 15-2A (580 mg, 1.86 mmol), bis(pinacolato)diboron (505.16 mg, 2.24 mmol) were dissolved in toluene (10 mL), then potassium acetate (457.26 mg, 4.66 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium chloride (136.37 mg, 186.37 μmol) were added, the reaction was stirred at 80 °C for 2 hr under nitrogen. Then the reaction was filtered to remove insoluble impurities, the filtrate was concentrated to dryness to give compound 15-3A. LCMS: m / z = 345.1 [M+1] + .

[0647] Step 3

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

[0649] Step 4

[0650] Referring to the synthetic method of Reference Examples 6-9, compound 15A was prepared. LCMS: m / z = 834.5 [M+H] + .

[0651] Example 16

[0652]

[0653]

[0654] Referring to the synthetic method of Reference Examples 6-9, 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] + ) were prepared.

[0655] Biological test data

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

[0657] Experimental materials:

[0658] RPMI 1640 medium and penicillin / streptomycin antibiotics were purchased from Gibco, fetal bovine serum was purchased from Hyclone, 3D CellTiter-Glo (Cell Viability Chemiluminescent Assay) reagent was purchased from Promega, and AsPC-1 cell line was purchased from ATCC, Envision Multi-label Reader (PerkinElmer).

[0659] Experimental method:

[0660] AsPC-1 cells were seeded in ultra-low attachment 96-well U-bottom plates at 80 μL cell suspension per well, containing 1000 AsPC-1 cells. The cell plates were incubated in a carbon dioxide incubator overnight.

[0661] The test compounds were diluted by 5 times with an 8-point dilution series, from 2 mM to 25.6 nM, and a duplicate well assay was set up. 78 μL of medium was added to the middle plate, and then 2 μL of each compound was transferred to the middle plate according to the corresponding position, and 20 μL per well was transferred to the cell plate after mixing. The concentration range of the compound transferred to the cell plate was 10 μM to 0.128 nM. The cell plate was incubated in a carbon dioxide incubator for 10 days. Another cell plate was prepared, and the signal value was read on the day of drug addition as the maximum value (Max value in the equation below) for data analysis.

[0662] The cell viability chemiluminescent assay reagent was added to the cell plate at 100 μL per well, and the luminescent signal was allowed to stabilize at room temperature for 30 minutes. The multi-label reader was used to read the signal.

[0663] Data analysis:

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

[0665] Table 1: In vitro screening test results of the compounds of the present application

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

[0667] Conclusion: The compound of the present application has significant inhibitory activity on AsPC-1 cell proliferation.

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

[0669] Experimental materials:

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

[0671] 3D CellTiter-Glo (Cell Viability Chemiluminescent Assay) 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 cell (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), NCI-H727 cell (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), A427 cell line (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), Capan-1 cell (IMDM + 20% FBS + 1% penicillin / streptomycin) were purchased from Nanjing Kebai Biological Technology Co., Ltd.; A375 cell line (DMEM + 10% FBS + 1% penicillin / streptomycin), AsPC-1 cell (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), PSN-1 cell line (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), SW620 cell (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), HCT116 cell (RPMI1640 + 10% FBS + 1% penicillin / streptomycin), A549 cell (F12K + 10% FBS + 1% penicillin / streptomycin), NCI-H441 cell (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 method:

[0673] The cells were seeded in ultra-low adsorption 96-well U-shaped plates, 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 test compounds were diluted 5-fold in 8 concentrations, from 2 mM to 25.6 nM, using a gun. A double-replicate experiment was set up. 78 μL of medium was added to the middle plate, and then 2 μL of each well of the gradient-diluted compound was transferred to the middle plate according to the corresponding position. After mixing, 20 μL of each well was transferred to the cell plate. The compound concentration range transferred to the cell plate was 10 μM to 0.128 nM. The cell plate was incubated in a carbon dioxide incubator for 5 days. Another cell plate was prepared, and the signal value was read on the day of drug addition as the maximum value (Max value in the following equation) for data analysis.

[0675] 50 μL of cell viability chemiluminescence detection reagent was added to each well of the cell plate, and the luminescence signal was allowed to stabilize by incubation at room temperature for 30 minutes. A multi-label analyzer was used for reading.

[0676] Data analysis:

[0677] The original data was converted into inhibition rate using the equation (Sample-Min) / (Max-Min)*100%, and the IC value was obtained by four-parameter curve fitting (obtained in the "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 application on cell proliferation. 50

[0678] Table 2: Experimental results of the in vitro cell proliferation inhibitory activity of the compounds of the present application

[0679]

[0680]

[0681] Conclusion: The compounds of the present application 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 exhibit obvious inhibitory effect in wild-type non-dependent cell lines (such as A375), and have good selectivity.

[0682] Experimental Example 3: Detection of p-ERK level 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; the components of ERK1 / 2 (THR202 / TYR204) KIT were: Advanced PhosphoERK1 / 2 Eu Cryptate antibody, Advanced PhosphoERK1 / 2 d2 antibody, Blocking reagent (stock solution 100X), Lysis buffer #1 (stock solution 4X), Detection buffer (ready-to-use), and the storage temperature was ≤-16℃.

[0685] Experimental method:

[0686] (1) Cells were seeded in white-bottom 384-well cell culture plates, 8 μL cell suspension per well, each well containing 7500 cells, and the cell plate was placed in a carbon dioxide incubator at 37 degrees overnight for incubation;

[0687] (2) The test compound was diluted to 3 mM with 100% DMSO as the first concentration, and then diluted to 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, 0.1 μM ten concentrations with a pipette. Take 2 μL compound and add to 198 μL cell starvation medium, mix well, then take 15 μL compound solution and add to 35 μL cell starvation medium and mix well, then add the last step of compound solution to the corresponding cell plate hole with 4 μL per hole, and the cell plate is placed back into the carbon dioxide incubator for further incubation for 3 hours, at which time the compound concentration is 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, 0.1 nM;

[0688] (3) After the incubation was completed, 3 μL of 5X cell lysis solution was added to each well, and the plate was shaken at room temperature for 30 minutes;

[0689] (4) The Phospho-ERK1 / 2 Eu Cryptate antibody and Phospho-ERK1 / 2 d2 antibody were diluted 20 times with Detection buffer and mixed at a ratio of 1:1, and 5 μL was added to each well of the cell culture plate, and incubated at room temperature for 2 hours;

[0690] (5) After incubation, read HTRF excitation: 320 nm, emission: 615 nm, 665 nm using a multilabel reader.

[0691] Data analysis:

[0692] Raw data was converted to % inhibition using the equation (Sample-Min) / (Max-Min)*100% and IC 50 values were obtained by four parameter curve fitting (GraphPad Prism, log(inhibitor) vs. response - Variable slope mode). Table 4 provides the p-ERK inhibition of the compounds of the present application. Max wells: positive control wells read at IX lysis; Min wells: negative control wells read at 0.5% DMSO cell lysate. The results of the experiment are shown in Table 3.

[0693] Table 3: Results of in vitro pERK inhibition screening assay for compounds of the application

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

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

[0696] Experimental Example 4: 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; the components of the Advanced Phospho-ERK1 / 2 (THR202 / TYR204) KIT were: Advanced PhosphoERK1 / 2 Eu Cryptate antibody, Advanced PhosphoERK1 / 2 d2 antibody, Blocking reagent (stock solution 100X), Lysis buffer #1 (stock solution 4X), Detection buffer (ready-to-use), and the storage temperature was ≤ -16°C.

[0699] Experimental method:

[0700] (1) Cells were seeded in white-bottom 384-well cell culture plates, 8 μL cell suspension per well, each well contained 7500 cells, the cell plate was put into a carbon dioxide incubator, 37 degrees overnight incubation;

[0701] (2) The test compound was diluted to 3 mM with 100% DMSO as the first concentration, and then diluted to 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, 0.1 μM ten concentrations with a pipette. Take 2 μL compound and add to 198 μL cell starvation medium, mix well, then take 15 μL compound solution and add to 35 μL cell starvation medium and mix well, then add the last step of compound solution to the corresponding cell plate well with 4 μL per well, the cell plate was put back into the carbon dioxide incubator for 3 hours of incubation, at this time the compound concentration was 3000, 1000, 300, 100, 30, 10, 3, 1, 0.3, 0.1 nM;

[0702] (3) After the incubation was completed, 3 μL of 5X cell lysis solution was added to each well, and incubated at room temperature for 30 minutes with shaking;

[0703] (4) Phospho-ERK1 / 2Eu Cryptate antibody and Phospho-ERK1 / 2d2 antibody were diluted 20 times with Detection buffer and mixed at a ratio of 1:1, 5 μL per well was added to the cell culture plate, and incubated at room temperature for 2 h;

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

[0705] Data analysis:

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

[0707] Table 4: In vitro pERK inhibition screening test results of the compounds of the present application

[0708]

[0709]

[0710] Conclusion: The compound of the present application has significant inhibitory activity on the pERK level of GP2D cells.

[0711] Experimental Example 5: In vivo pharmacodynamic study

[0712] Experimental purposes:

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

[0714] Experimental methods and steps:

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

[0716] (1) Cell culture: human lung bronchial benign tumor cells were cultured in vitro in monolayer, and the culture conditions were as follows: Gibco RMPI1640 medium containing 10% fetal bovine serum, 37°C, 5% CO2 incubator. Routine digestion treatment was carried out once a week with trypsin-EDTA. When the cell saturation was 80%-90% and the number reached the requirement, the cells were collected, counted and inoculated.

[0717] (2) Tumor cell inoculation and grouping: 0.2 mL (2×10 6 NCI-H727 cells (mixed with Matrigel, volume ratio 1:1) were inoculated subcutaneously in the right back of each mouse, and the average tumor volume reached about 134 mm 3 When the average tumor volume reached about 134 mm

[0718] The solvent was 5% DMSO / 10% solutol / 85% water. Control group: 10 μL / g of solvent was administered by gavage twice a day; treatment group: the test compound was dissolved in the solvent, and the compound was administered by gavage once a day, the dose is shown in Table 4.

[0719] (3) The tumor diameter was measured twice a week with a vernier caliper, and the tumor volume (V) was calculated according to the formula: V = 0.5a×b 2 , where a and b are the long diameter and short diameter of the tumor, respectively. The antitumor effect of the test compound was evaluated by tumor growth inhibition rate (TGI). The calculation formula is: TGI (%) = [1–(the average tumor volume at the end of treatment in a certain treatment group - the average tumor volume at the beginning of treatment in that treatment group) / (the average tumor volume at the end of treatment in the solvent control group - the 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 experimental results are shown in Table 5.

[0722] Table 5: Evaluation of the antitumor effect of the compound of the present application in the human lung bronchial benign tumor NCI-H727 model

[0723]

[0724]

[0725] Conclusion: The compound of the present application exhibits excellent antitumor effect in the NCI-H727 tumor model.

[0726] Experimental Example 6: In vivo pharmacodynamics study

[0727] Experimental purpose:

[0728] The in vivo efficacy of the compound of the present application in the human pancreatic cancer PK59 cell BALB / c nude mouse subcutaneous xenograft tumor model was studied.

[0729] Experimental methods and steps:

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

[0731] (1) Cell culture: human pancreatic cancer PK59 cells were cultured in vitro in a monolayer, and the culture conditions were as follows: Gibco RMPI1640 medium containing 10% fetal bovine serum, 37°C, 5% CO2 incubator. The cells were routinely digested and passaged with trypsin-EDTA once a week. When the cell saturation was 80%-90% and the number reached the requirement, the cells were collected, counted, and inoculated.

[0732] (2) Tumor cell inoculation and grouping: 0.2 mL (2×10 6 PK59 cells (mixed with Matrigel at a volume ratio of 1:1) were inoculated subcutaneously on the right back of each mouse. When the average tumor volume reached about 104.4 mm 3 , the mice were grouped and administered, with 6 animals in each group.

[0733] The vehicle was 5% DMSO / 10% solutol / 85% water. Control group: once a day, intragastrically administered with the vehicle at a dose of 10 μL / g; treatment group: after dissolving the test compound in the vehicle, once a day, intragastrically administered with the compound, and the dose is shown in Table 5.

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

[0735] Experimental results:

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

[0737] Table 6 Experimental results of the antitumor efficacy of the compound of the present application in the human pancreatic cancer PK59 model

[0738]

[0739]

[0740] Conclusion: The compound of the present application exhibits excellent antitumor effect in the PK59 tumor model.

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

[0742] Experimental procedure:

[0743] (1) Collect fresh mouse and human whole blood (the number of animal individuals is required to be n≥3, and the number of human individuals is required to be n≥2) with blood collection tubes containing EDTA K2 anticoagulant. The whole blood sample should be mixed uniformly before use and stored at 2-8°C or on wet ice. The whole blood should be used within 36 hours after collection. The blank whole blood is centrifuged at 2000 x g for 15 minutes at room temperature to obtain blank plasma. Check the plasma state, and the plasma sample with hemolysis cannot be used.

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

[0745] (3) Diclofenac (for all species), chlorthalidone (for mice) or chloroquine (for humans) are used as control compounds.

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

[0747] (5) Incubate the whole blood sample containing the test compound at 37°C for 60 minutes.

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

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

[0750] (8) When the samples are treated, all the samples are subjected to matrix matching (i.e. adding the same volume of blank whole blood or blank plasma), and mixing well.

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

[0752] (10) The analyte and control compounds in the sample are determined by liquid chromatography-tandem mass spectrometry (LC-MS / MS) method. The ratio of the peak area of the analyte to the peak area of the internal standard is used to represent 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 the whole blood and the plasma, K E / P represents the concentration ratio of the compound in the red blood cell and the plasma, and the recovery rate (%) represents the recovery rate of the compound in the whole blood.

[0755] Table 7 Distribution results of the compound of the present application in whole blood and plasma

[0756]

[0757]

[0758] Conclusion: Compared with human and mouse plasma, the compound of the present application has higher distribution in whole blood and red blood cells.

[0759] Experimental Example 8: Pharmacokinetic test in mice

[0760] Purpose of the experiment:

[0761] To evaluate the pharmacokinetic behavior of the compound of the present application in male CD-I (ICR) mice.

[0762] Method of the experiment:

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

[0764] Results of the experiment:

[0765] In the experiment, all animals were well tolerated and no abnormal behavior was observed. After intravenous bolus administration of compound 7A, the whole blood clearance (Cl) was 6.29 mL / min / kg, the steady state volume of distribution (Vd) was 2.18 L / kg, the elimination half-life (T 1 / 2 ) was 4.29 h, and the area under the whole blood concentration-time curve (AUC 0-last ) from time zero to the last quantifiable time point was 3160 h-nmol / mL. After oral gavage of compound 7A, the time to peak concentration (T max ) occurred at 3.0 h post-dose, the peak concentration (C max ) was 2000 nmol / mL, the AUC 0-last was 17119 h-nmol / mL, and the bioavailability (F) was 54.3%.

[0766] Conclusion: The compound of the present application has a high exposure, a long half-life, and good pharmacokinetic properties.

[0767] Experimental Example 9: Pharmacokinetic study in rats

[0768] Purpose of the experiment:

[0769] To evaluate the pharmacokinetic behavior of the compound of the present application in male SD rats.

[0770] Method of the experiment:

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

[0772] Results of the experiment:

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

[0774] Table 8. Results of rat pharmacokinetic test of the compound of the present application

[0775]

[0776]

[0777] Conclusion: The compound of the present application has high exposure, long half-life and good pharmacokinetic properties.

[0778] Experimental Example 10: Pharmacokinetic study in beagle dogs

[0779] Purpose of the experiment:

[0780] To evaluate the pharmacokinetic behavior of the compound of the present application in male beagle dogs.

[0781] Method of the experiment:

[0782] The test compound was dissolved in solvent (5% DMSO / 10% solutol / 85% water). Four male beagle dogs were divided into two groups, two in each group. The first group of animals was administered the compound by single intravenous bolus (i.v.) at a dose of 1 mg / kg. The second group of animals was administered the compound by single oral gavage (p.o.) at a dose of 5 mg / kg. Whole blood samples were collected at 0.083 (only i.v. bolus group), 0.25, 0.5, 1, 2, 4, 8 and 24 hours post-dose. The concentration of the test compound in the whole blood samples was determined using the LC-MS / MS method.

[0783] Results of the experiment:

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

[0785] Table 9. Results of beagle dog pharmacokinetic test of the compound of the present application

[0786]

[0787] Conclusion: The compound of the present application has 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; L2 is L3is selected from -CH2-, To Ring B is R2 is selected from -0- and -NH-; R3is selected from phenyl and 5-6 membered heteroaryl, each independently optionally substituted with 1, 2, or 3 R b substituents; R6is selected from C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl, said C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R d substituents; each R7is independently selected from H, halo, C 1-4 alkyl and C 1-4 alkoxy, said C 1-4 alkyl and C 1-4 alkoxy is independently optionally substituted with 1, 2, or 3 R e substituents; R8is selected from C 3-6 cycloalkyl and 3-7 membered heterocycloalkyl, said C 3-6 cycloalkyl and 3-7 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R f substituents; each R b , each R e , and each R f is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3, and OCF3; each R d are each independently C 1-4 alkyl, said C 1-4 alkyl is optionally substituted with 1, 2, or 3 R; each R is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3, and OCF3; p is 1; q is selected from 0, 1, 2, and 3; the "3-6 membered heterocycloalkyl", "3-7 membered heterocycloalkyl", and "5-6 membered heteroaryl" each independently contains 1 or 2 heteroatoms independently selected from -NH-, -0-, -S-, and N.

2. A compound represented by Formula (III), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, L is R6; L2 is L3is selected from -CH2-, To Ring B is R2 is selected from -0- and -NH-; R3is selected from phenyl and 5-6 membered heteroaryl, each independently optionally substituted with 1, 2, or 3 R b substituents; R6is selected from C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl, said C 3-6 cycloalkyl and 3-6 membered heterocycloalkyl are each independently optionally substituted with 1, 2, or 3 R d substituents; each R7is independently selected from H, halo, C 1-4 alkyl and C 1-4 alkoxy, said C 1-4 alkyl and C 1-4 alkoxy is independently optionally substituted with 1, 2, or 3 R e substituents; To each R b and each R e is independently selected from F, CI, Br, I, OH, CH3, CF3, OCH3, and OCF3; Each R d C 1-4 Alkyl, the C 1-4 The alkyl group is optionally substituted with 1, 2 or 3 R groups; each R is independently selected from F, Cl, Br, I, OH, CH3, CF3, OCH3, and OCF3; q is selected from 0, 1, 2, and 3; the "3-6 membered heterocycloalkyl", "3-7 membered heterocycloalkyl", and "5-6 membered heteroaryl" each independently contains 1 or 2 heteroatoms independently selected from -NH-, -0-, -S-, and N.

3. The compound according to claim 1 or 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R6is selected from cyclopropyl, cyclobutyl, cyclopentyl, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, piperazinyl, morpholinyl, oxetanyl, oxetanyl, and oxepanyl, each independently optionally substituted with 1, 2, or 3 R d substituents.

4. The compound according to claim 1 or 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R6is selected from 5. The compound according to claim 1 or 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R6 is 6. The compound according to claim 1 or 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, L is selected from 7. The compound according to claim 1 or 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, L is 8. The compound according to claim 1 or 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R3is selected from phenyl, pyridyl, pyrimidinyl, thiazolyl, oxazolyl, pyrazolyl, and imidazolyl, each of which is independently optionally substituted with 1, 2, or 3 R b substituents.

9. The compound according to claim 1 or 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, R3is selected from 10. The compound according to claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R8is selected from the group consisting of piperazinyl, homopiperazinyl, piperidinyl, homopiperidinyl, and morpholinyl, each of which is independently optionally substituted with 1, 2, or 3 R f substituted.

11. The compound according to claim 1, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R8 is 12. The compound according to claim 2, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, wherein, Structural unit To 13. The compound according to claim 1 or 2, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R2 is -0-.

14. The compound according to claim 1 or 2, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, R7 is selected from H, F, Cl, CH3, and CH2CH3.

15. The compound according to claim 1 or 2, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, each R is independently selected from the group consisting of CH3and CH2CH3, said CH3and CH2CH3are each independently optionally substituted with 1, 2, or 3 R. d each R is independently selected from the group consisting of CH3and CH2CH3, said CH3and CH2CH3are each independently optionally substituted with 1, 2, or 3 R.

16. The compound according to claim 1 or 2, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, wherein, Each R d are each independently CH3.

17. A compound represented by Formula (III), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, 18. A compound represented by Formula (III), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, 19. A compound represented by Formula (III), a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, 20. A compound represented by Formula (III), or a pharmaceutically acceptable salt thereof,

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