Indole compound, indole dimer compound and application of indole compound and indole dimer compound as STING agonist

Abstract

The invention provides a compound, which is a compound as shown in formula (I) or a stereoisomer, a tautomer, a solvate and a pharmaceutically acceptable salt of the compound as shown in formula (I), and application of the compound as an STING agonist and in prevention or treatment of cell proliferation diseases, especially in prevention or treatment of tumors and cancers # imgabs0 #

Description

[0001] Priority information

[0002] This application claims priority to and the benefits of patent application 202311071768.3 filed on August 23, 2023, and is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention belongs to the field of biopharmaceutical technology. Specifically, the present invention relates to an indole compound and an indole dimer compound and their use as a STING agonist. More specifically, the present invention relates to a compound, a dimer compound, a STING agonist, a pharmaceutical composition and their use. Background Art

[0004] Stimulator of interferon genes (STING, also known as transmembrane protein 173 / TMEM173 / MPYS / MITA / ERIS) is a signaling molecule encoded by the TMEM173 gene in the human body. STING is a protein with 379 amino acids, consisting of several transmembrane regions. STING protein is expressed in several endothelial and epithelial cell types, as well as hematopoietic lineages such as T cells, dendritic cells (DCs) and macrophages. STING is associated with the endoplasmic reticulum (ER) in cells and plays a major role in controlling the transcription of many host defense genes, including type I interferons (IFNs) and proinflammatory cytokines.

[0005] Tumor immunotherapy has gradually developed into an important and highly promising direction for cancer treatment. In immunotherapy, how to effectively enhance the immune response of the tumor microenvironment and alleviate immune tolerance is a core issue related to the treatment effect. As a new pattern recognition receptor recently discovered, the stimulator of interferon genes (STING) pathway has attracted the research and development interest of many large pharmaceutical companies. Therefore, the development of an effective STING pathway agonist for clinical tumor immunotherapy has important research value. Summary of the invention

[0006] In the first aspect of the present invention, the present invention provides a compound, which is a compound represented by formula (I) or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the compound represented by formula (I):

[0007]

[0008] Wherein, R7 and R8 are each independently selected from H, halogen, -CN, -OR 14 、-NHR 14 or -R 14; or, R7 and R8 are linked together with the atoms to form a 4-7 membered heterocyclic ring, wherein the 4-7 membered heterocyclic ring contains at least one heteroatom selected from O, N and S; or one of R7 and R8 is linked to A and connected;

[0009] R7' and R8' are each independently selected from H, halogen, -CN, -OR 14 、-NHR 14 or -R 14 ;

[0010] A is empty or -C 1～6 Alkylene-, wherein the -C 1～6 Alkylene - optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NHC(O)-C 1～3 Alkyl, -NO2 or -CN substitution;

[0011] R1 and R1' are each independently empty, optionally substituted with one or more R9 -C 1～6 Alkylene-, -C(O)-C optionally substituted with one or more R9 1～6 Alkylene-, -C(NOH)-C optionally substituted with one or more R9 2～5 alkylene, -C(O)-C optionally substituted by one or more R9 1～6 Alkyleneoxy-, -C(O)-C optionally substituted by one or more R9 1～6 Cycloalkylene-, -C(O)-C optionally substituted by one or more R9 2～6 Alkenylene-, -C(O)-C optionally substituted with one or more R9 2～6 Alkynylidene- or -C(O)-NR 10 R 11 -;

[0012] R2 and R2' are each independently -CN, -C(O)-, -C(O)R 12 、-C(O)N(R 12 ) a 2- or 4- to 7-membered heterocyclic ring, wherein the 4- to 7-membered heterocyclic ring contains at least one heteroatom selected from O, N and S;

[0013] R3, R3', R4 and R4' are each independently H, halogen, optionally substituted with one or more R 13 Replaced-C 1～6 Alkyl, optionally with one or more R 13 Replaced-C 1～6 Alkoxy, optionally with one or more R 13 Replaced-C 2～6 alkenyl, optionally substituted with one or more R13 Replaced-C 2～6 Alkynyl, optionally substituted with one or more R 13 Substituted -C(O)-C 1～6 Alkyl, optionally with one or more R 13 Substituted -C(O)-C 1～6 Alkoxy, optionally with one or more R 13 Substituted -C(O)-C 2～6 alkenyl, optionally substituted with one or more R 13 Substituted -C(O)-C 2～6 Alkynyl, optionally substituted with one or more R 13 Replaced-C 0～6 Alkyl-C(O)OH, optionally with one or more R 13 Replaced-C 1～6 Alkoxy-C(O)OH, optionally with one or more R 13 Replaced-C 2～6 Alkenyl -C(O)OH or optionally substituted by one or more R 13 Replaced-C 2～6 Alkynyl-C(O)OH;

[0014] R5, R5', R6 and R6' are each independently H, halogen, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 1～6 Cycloalkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl;

[0015] Each R9 is independently selected from halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl or -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0016] Each R 10 Independently selected from H, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OR 15, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0017] Each R 11 independently selected from H, halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution;

[0018] Each R 12 Independently selected from H, -OR 15 , -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, -C 5～7 Aryl or 4 to 7 membered ring, wherein the -OR 15 , -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 5～7 Aryl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)-, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution;

[0019] Each R 13 independently selected from halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, -C 1～6 Alkyl and -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0020] Each R 14are independently selected from H, -Bn, halogen, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkenyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0021] Each R 15 Independently selected from H or -C 1～3 alkyl;

[0022] X1 and X2 are each independently C or N, and X1 and X2 are not C at the same time;

[0023] X3, X4 and X5 are each independently C or N; or one of X1 and X2, X3 and R1 together with the connected atoms are linked to form a 4-7 membered heterocyclic ring, and the 4-7 membered heterocyclic ring contains N or Se.

[0024] The compound represented by formula (I) according to the embodiments of the present invention, or its stereoisomer, or its tautomer, or its solvate, or its pharmaceutically acceptable salt can effectively activate the STING protein, and can be used as a STING agonist to effectively prevent or treat cell proliferation diseases, such as tumors and cancer.

[0025] According to an embodiment of the present invention, R1 and R1' are each independently -C optionally substituted by one or more R9 1～6 Alkylene-, -C(O)-C optionally substituted with one or more R9 1～6 alkylene, -C(O)-C optionally substituted by one or more R9 1～6 Cycloalkylene or -C(O)-NR 10 R 11 -.

[0026] According to an embodiment of the present invention, each R9 is independently selected from halogen, -C 1～3 Alkyl and -C 1～3 Alkoxy, wherein the -C 1～3 Alkyl and -C 1～3 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution.

[0027] In some preferred embodiments of the present invention, each R9 is independently selected from halogen, -C 1～3 Alkyl or -C 1～3 Alkoxy.

[0028] According to an embodiment of the present invention, each R 10 Independently selected from -C 1～4 Alkyl, wherein the -C 1～4 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2, -CN or -C 1～2 Alkyl substitution.

[0029] According to an embodiment of the present invention, each R 11 are independently selected from H, halogen, -C 1～4 Alkyl, wherein the -C 1～4 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

[0030] According to an embodiment of the present invention, R1 and R1' are each independently -C(O)-NR 10 R 11 -, one of X1 and X2, X3 and the N of R1 are linked together with the atoms connected thereto to form a 4- to 7-membered heterocyclic ring.

[0031] According to an embodiment of the present invention, R2 is -C(O)R 12 、-C(O)N(R 12 ) a 2- or 4- to 7-membered heterocyclic ring, wherein the 4- to 7-membered heterocyclic ring contains at least one heteroatom selected from O, N and S.

[0032] According to an embodiment of the present invention, each R 12 Independently selected from H, -OR 15 or -C 1～3 Alkyl, wherein the -OR 15 or -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN or phenyl.

[0033] According to an embodiment of the present invention, each R 15 Independently selected from H or -C 1～3 alkyl.

[0034] According to an embodiment of the present invention, each R 13 Independently selected from -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, or -CN.

[0035] According to an embodiment of the present invention, R3 and R4 are each independently H, halogen, -C optionally substituted by one or more -OH 1～3 Alkyl or -C 0～2 Alkyl-C(O)OH.

[0036] According to an embodiment of the present invention, R5 and R6 are each independently H, halogen or -C 1～3 alkyl.

[0037] According to an embodiment of the present invention, R7 and R8 are each independently -OR 14 ; Or, R7 and R8 are linked together with the atoms to which they are connected to form a 4- to 7-membered heterocyclic ring containing at least one heteroatom selected from O, N and S.

[0038] According to an embodiment of the present invention, each R 14 are independently selected from H, -C optionally substituted with one or more halogens 1～3 Alkyl or -C optionally substituted by one or more halogen 2～3 Alkenyl.

[0039] According to an embodiment of the present invention, X1 is C, X2 is N or Se, or X1 is N or Se, and X2 is C.

[0040] According to an embodiment of the present invention, X3, X4 and X5 are C.

[0041] According to an embodiment of the present invention, the compound is a compound represented by formula (Ia) or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the compound represented by formula (Ia):

[0042]

[0043] Wherein, R1 is -C(OH)-C 1～5 Alkylene, -C(O)-C 2～5 Alkylene, -C(O)-C 2～4 Cycloalkylene or -C(O)-NR 10 R 11 -;

[0044] R 10 is H or -CH3;

[0045] R 11 For -C 2～4 alkyl;

[0046] R2 is -C(O)OH, -C(O)OCH3, -C(O)OCH2CH3, -C(O)N(H)OCH3, -C(O)N(H)(CH2)2OH or

[0047] R3 is H, -CH3, -CH2CH3, -C(O)CH3, -C 1～5 Alkylene -C(O)OH or -C(O)OH;

[0048] R4, R5 and R6 are each independently H or halogen;

[0049] R7 and R8 are each independently -OH, -OCH3 optionally substituted by halogen, or -OCH2CH=CH2.

[0050] According to an embodiment of the present invention, -R1-R2 has the following structure:

[0051]

[0052]

[0053] and -C(O)OH.

[0054] According to an embodiment of the present invention, the compound is a stereoisomer, a tautomer, a solvate, or a pharmaceutically acceptable salt of the compound represented by formula (VI):

[0055]

[0056] Among them, R 1a is hydrogen or -CH3;

[0057] R 1b and R 1c are each independently H or halogen;

[0058] R 1d is -OH, -OCH3 or -OCH2CH3;

[0059] R 1f is -CH3 or -CH2-CH=CH2;

[0060] R 1g for

[0061] Among them, R 1f for When R 1d Not -OH.

[0062] In the second aspect of the present invention, the present invention provides a compound, which is a compound shown in the following structure or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the compound shown in the following structure:

[0063]

[0064]

[0065]

[0066]

[0067] The compounds according to the embodiments of the present invention, or their stereoisomers, or their tautomers, or their solvates, or their pharmaceutically acceptable salts can effectively activate the STING protein, and can be used as STING agonists to effectively prevent or treat cell proliferation diseases, such as tumors and cancers.

[0068] In the third aspect of the present invention, the present invention provides a dimer compound, which is a dimer compound represented by formula (II), formula (III) or formula (IV), or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the dimer compound represented by formula (II), formula (III) or formula (IV):

[0069]

[0070] Where A is empty or -C 1～6 Alkylene-, wherein the -C 1～6 Alkylene - optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NHC(O)-C 1～3 Alkyl, -NO2 or -CN substitution;

[0071] R1 and R1' are each independently empty, optionally substituted with one or more R9 -C(O)-C 1～6 Alkylene-, -C(NOH)-C optionally substituted with one or more R9 1～6 alkylene, -C(NOAc)-C optionally substituted with one or more R9 2～5 alkylene, -C(O)-C optionally substituted by one or more R9 1～6 Alkyleneoxy-, -C(O)-C optionally substituted by one or more R9 1～6 Cycloalkylene-, -C(O)-C optionally substituted by one or more R9 2～6 Alkenylene-, -C(O)-C optionally substituted with one or more R9 2～6 Alkynylidene- or -C(O)-NR 10 R 11 -;

[0072] R2 and R2' are each independently -CN, -C(O)-, -C(O)R 12、-C(O)N(R 12 ) a 2- or 4- to 7-membered heterocyclic ring, wherein the 4- to 7-membered heterocyclic ring contains at least one heteroatom selected from O, N and S;

[0073] R3, R3', R4 and R4' are each independently H, halogen, optionally substituted with one or more R 13 Replaced-C 1～6 Alkyl, optionally with one or more R 13 Replaced-C 1～6 Alkoxy, optionally with one or more R 13 Replaced-C 2～6 alkenyl, optionally substituted with one or more R 13 Replaced-C 2～6 Alkynyl, optionally substituted with one or more R 13 Substituted -C(O)-C 1～6 Alkyl, optionally with one or more R 13 Substituted -C(O)-C 1～6 Alkoxy, optionally with one or more R 13 Substituted -C(O)-C 2～6 alkenyl, optionally substituted with one or more R 13 Substituted -C(O)-C 2～6 Alkynyl, optionally substituted with one or more R 13 Replaced-C 0～6 Alkyl-C(O)OH, optionally with one or more R 13 Replaced-C 1～6 Alkoxy-C(O)OH, optionally with one or more R 13 Replaced-C 2～6 Alkenyl -C(O)OH or optionally substituted by one or more R 13 Replaced-C 2～6 Alkynyl-C(O)OH;

[0074] R5, R5', R6 and R6' are each independently H, halogen, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 1～6 Cycloalkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl;

[0075] R7, R7', R8 and R8' are each independently H, halogen, -CN, -OR 14 、-NHR 14 or -R 14 ;

[0076] Each R9 is independently selected from H, halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C1～6 Alkyl or -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0077] Each R 10 Independently selected from H, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OR 15 , -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0078] Each R 11 independently selected from H, halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution;

[0079] Each R 12 Independently selected from H, -OR 15 , -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, -C 5～7 Aryl or 4 to 7 membered ring, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 5～7 Aryl or -C 2～6The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)-, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution;

[0080] Each R 13 independently selected from halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, -C 1～6 Alkyl and -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0081] Each R 14 are independently selected from H, -Bn, halogen, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkenyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution;

[0082] Each R 15 Independently selected from H or -C 1～3 alkyl;

[0083] X1 and X2 are each independently C, N, S or Se, and X1 and X2 are not C at the same time;

[0084] X3, X4 and X5 are each independently C or N.

[0085] According to the embodiments of the present invention, the dimer compound represented by formula (II), formula (III) or formula (IV), or its stereoisomer, or its tautomer, or its solvate, or its pharmaceutically acceptable salt can effectively activate the STING protein, and can be used as a STING agonist to effectively prevent or treat cell proliferation diseases, such as tumors and cancer.

[0086] According to an embodiment of the present invention, R1 and R1' are each independently -C(O)-C optionally substituted by one or more R9. 1～6 Alkylene- or -C(O)-NR10 R 11 -.

[0087] According to an embodiment of the present invention, each R9 is independently selected from halogen and -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

[0088] According to an embodiment of the present invention, each R 10 Independently selected from -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

[0089] According to an embodiment of the present invention, each R 11 Independently selected from H or -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

[0090] According to an embodiment of the present invention, R2 and R2' are each independently -C(O)R 12 or -C(O)N(R 12 )2.

[0091] According to an embodiment of the present invention, each R 12 Independently selected from H, -OR 15 or -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

[0092] According to an embodiment of the present invention, each R 15 Independently selected from H or -C 1～3 alkyl.

[0093] According to an embodiment of the present invention, R3, R3', R4, R4', R5, R5', R6 and R6' are each independently H, halogen or -C 1～3 alkyl.

[0094] According to an embodiment of the present invention, R7, R7', R8 and R8' are each independently -C 1～3 Alkoxy.

[0095] According to an embodiment of the present invention, A is empty or -C 1～4 Alkylene, wherein the -C 1～4Alkylene - optionally substituted with one or more halogen, -OH, -SH, -NH2, -NHC(O)-C 1～3 Alkyl, -NO2 or -CN substituted.

[0096] According to an embodiment of the present invention, the dimer compound is a dimer compound represented by formula (V), or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the dimer compound represented by formula (V):

[0097]

[0098] Among them, A is

[0099] R 2a and R 2b Each independently And R 2a and R 2b Not containing N at the same time;

[0100] R 2c and R 2d Each is independently -OH, -NH2, -OCH3, -OCH2CH3, -OCH2(CH3)2, -OCH2CH2(CH3)2 or C 5～7 Aryloxy.

[0101] According to an embodiment of the present invention, R 2c and R 2d Each is independently -OH, -OCH3 or -OCH2CH3.

[0102] In the fourth aspect of the present invention, the present invention provides a dimer compound, which is a dimer compound shown in the following structure or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the dimer compound shown in the following structure:

[0103]

[0104]

[0105] The dimer compound according to the embodiments of the present invention, or its stereoisomer, or its tautomer, or its solvate, or its pharmaceutically acceptable salt can effectively activate the STING protein, and can be used as a STING agonist to effectively prevent or treat cell proliferation diseases, such as tumors and cancer.

[0106] In the fifth aspect of the present invention, the present invention proposes use of the compound described in the first aspect or the second aspect or the dimer compound described in the third aspect or the fourth aspect as a STING agonist.

[0107] In the sixth aspect of the present invention, the present invention provides a STING agonist, which includes the compound described in the first aspect or the second aspect or the dimer compound described in the third aspect or the fourth aspect, and optionally a pharmaceutically acceptable carrier, excipient, or vehicle.

[0108] In the seventh aspect of the present invention, the present invention provides a pharmaceutical composition, which includes the compound described in the first aspect or the second aspect, the dimer compound described in the third aspect or the fourth aspect, or the STING agonist described in the sixth aspect.

[0109] According to an embodiment of the present invention, the pharmaceutical composition further includes pharmaceutically acceptable carriers, excipients, and vehicles.

[0110] In the eighth aspect of the present invention, the present invention proposes a use of the compound described in the first or second aspect, the dimer compound described in the third or fourth aspect, the STING agonist described in the sixth aspect, or the pharmaceutical composition described in the seventh aspect in the preparation of a drug for preventing or treating a cell proliferative disease.

[0111] According to an embodiment of the present invention, the cell proliferation disease is a tumor or cancer.

[0112] In the ninth aspect of the present invention, the present invention proposes a method for preventing or treating a cell proliferative disease, comprising: administering to a subject a pharmaceutically acceptable dose of the compound of the first aspect or the second aspect, the dimer compound of the third aspect or the fourth aspect, the STING agonist of the sixth aspect, or the pharmaceutical composition of the seventh aspect.

[0113] The effective amount of the compound, dimer compound, STING agonist or pharmaceutical composition of the present invention may vary depending on the mode of administration and the severity of the disease to be treated. The selection of the preferred effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials). The factors include, but are not limited to, the patient's age, weight, general health, sex, diet, administration time, drug combination, severity of the disease to be treated, route of administration, etc. For example, depending on the needs of the treatment situation, several divided doses may be administered daily, for example, four times a day, three times a day, twice a day, once a day, or once every other day, or several doses administered daily may be proportionally reduced; alternatively, the pharmaceutical composition or gel of the present invention is administered for at least one year or longer, preferably at least one month, more preferably at least one week, and most preferably at least one day, to achieve continuous relief of tumors or cancer.

[0114] The compounds, dimer compounds, STING agonists or pharmaceutical compositions of the present invention can be incorporated into suitable drugs, and these drugs can be prepared in various forms, such as liquid, semi-solid and solid dosage forms, including but not limited to solid dosage forms, semi-solid dosage forms, liquid dosage forms and gas dosage forms, etc. Various modes of administration of the compounds, dimer compounds, STING agonists, pharmaceutical compositions or drugs of the present invention are contemplated, including peritoneal, intravenous, intramuscular, subcutaneous, cortical, oral, topical, nasal, pulmonary, rectal and application, but the present invention is not limited to these exemplified modes of administration.

[0115] Additional aspects and advantages of the present invention will be given in part in the following description and in part will be obvious from the following description, or will be learned through practice of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0116] The above and / or additional aspects and advantages of the present invention will become apparent and easily understood from the description of the embodiments in conjunction with the following drawings, in which:

[0117] Figure 1 is the structural formula of the positive control compound diABZI of Test Example 1 of the present invention;

[0118] Figure 2 The weight change of mice after administration of compound 643 in Test Example 2 of the present invention;

[0119] Figure 3 The results of cytokine IFN-β in mouse serum samples in Test Example 2 of the present invention;

[0120] Figure 4 is the average drug concentration-time curve of compound 643 in mouse blood in Test Example 2 of the present invention; Figure 5 The weight change of mice after administration in Test Example 2 of the present invention; Figure 6 The results of detecting the cytokine IFN-β in the serum sample in Test Example 2 of the present invention; Figure 7 The average drug concentration-time curve of compound D-18 in mouse blood in Test Example 2 of the present invention; Figure 8 This is the average drug concentration-time curve of compound D-18 in test example 2 of the present invention in mouse tumors. DETAILED DESCRIPTION

[0121] The embodiments of the present invention are described in detail below. The embodiments described below are exemplary and are only used to explain the present invention, and should not be understood as limiting the present invention.

[0122] It should be noted that the terms "first" and "second" are used for descriptive purposes only and should not be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more of the features. Further, in the description of the present invention, unless otherwise specified, the meaning of "plurality" is two or more.

[0123] It should be noted that, for the structural formula and chemical formula description in the embodiments or embodiments of the present invention, the present invention is intended to cover all substitutions, modifications and equivalent technical solutions, which are all within the scope of the present invention as defined in the claims. Those skilled in the art will recognize that many methods and materials similar or equivalent to those described in the present invention can be used to practice the present invention. The present invention is by no means limited to the methods and materials described in the present invention. In the event that one or more of the combined documents, patents and similar materials are different from or contradictory to the present application (including but not limited to defined terms, term applications, described technologies, etc.), the present invention shall prevail.

[0124] It should be further appreciated that certain features of the invention, which for clarity are described in multiple separate embodiments or implementations, may also be provided in combination in a single embodiment or implementation. Conversely, various features of the invention, which for brevity are described in a single embodiment or implementation, may also be provided separately or in any suitable sub-combination.

[0125] Unless otherwise noted, technical and scientific terms used in the present invention have the same meanings as commonly understood by those skilled in the art to which the present invention belongs. Unless otherwise noted, all patent publications cited in the entire disclosure of the present invention are incorporated herein by reference in their entirety.

[0126] The following definitions shall apply to the present invention unless otherwise indicated. For the purposes of the present invention, chemical elements are defined according to the Periodic Table of the Elements, CAS version and Handbook of Chemicals, 75, th Ed, 1994. In addition, general principles of organic chemistry are found in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", by Michael B. Smith and Jerry March, John Wiley & Sons, New York: 2007, and all references are incorporated herein by reference.

[0127] In this document, the terms “include” or “comprising” are open expressions, that is, including the contents specified in the present invention but not excluding other contents.

[0128] As used herein, the compounds of the present invention also include isotopically labeled compounds of the present invention, which are identical to those described herein except for the fact that one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Exemplary isotopes that may also be incorporated into the compounds of the present invention include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as 2 H. 3 H. 13 C. 14 C. 15 N. 16 O. 17 O. 31 P. 32 P. 36 S. 18 F and 37 Cl.

[0129] Compounds of the invention containing the aforementioned isotopes and / or other isotopes of other atoms and pharmaceutically acceptable salts of the compounds are included within the scope of the invention. Isotopically labeled compounds of the invention, for example radioactive isotopes, such as 3 H and 14 C is incorporated into the compounds of the present invention for drug and / or substrate tissue distribution analysis. Due to the ease of preparation and detection, tritiated, i.e. 3 H, and carbon-14, i.e. 14 C, isotopes are particularly preferred. In addition, heavy isotopes, such as deuterium, 2 H substitution may offer certain therapeutic advantages resulting from greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and therefore may be preferred in some circumstances.

[0130] The stereochemical definitions and conventions used herein are generally in accordance with SP Parker, Ed., McGraw-Hill Dictionary of Chemical Terms (1984) McGraw-Hill Book Company, New York; and Eliel, E. and Wilen, S., "Stereochemistry of Organic Compounds", John Wiley & Sons, Inc., New York, 1994. The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers and mixtures thereof such as racemic mixtures, are also included within the scope of the present invention. Many organic compounds exist in optically active forms, i.e., they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefixes D and L or R and S are used to indicate the absolute configuration of the molecule with respect to the chiral center (or multiple chiral centers) in the molecule. The prefixes d and l or (+) and (-) are the signs used to designate the rotation of plane polarized light caused by a compound, where (-) or l indicates that the compound is levorotatory. A compound prefixed with (+) or d is dextrorotatory. With respect to a given chemical structure, these stereoisomers are identical except that they are mirror images of each other. Specific stereoisomers may also be referred to as enantiomers, and a mixture of such isomers is often referred to as a mixture of enantiomers. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may occur when there is no stereoselectivity or stereospecificity in a chemical reaction or process.

[0131] Depending on the choice of starting materials and processes, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, for example as pure optical isomers, or as isomer mixtures, such as racemic and diastereomeric mixtures, depending on the number of asymmetric carbon atoms. Optically active (R)- or (S)-isomers may be prepared using chiral synthons or chiral agents, or resolved using conventional techniques. If the compound contains a double bond, the substituents may be in the E or Z configuration; if the compound contains a disubstituted cycloalkyl group, the cycloalkyl substituents may be in the cis- or trans- configuration.

[0132] The compounds of the present invention may contain asymmetric centers or chiral centers and therefore exist in different stereoisomeric forms. It is contemplated that all stereoisomeric forms of the compounds of the present invention, including but not limited to diastereomers, enantiomers and atropisomers and geometric (or conformational) isomers and mixtures thereof, such as racemic mixtures, are within the scope of the present invention.

[0133] Unless otherwise indicated, structures depicted herein are also meant to include all isomers (e.g., enantiomers, diastereomeric atropisomers, and geometric (or conformational) forms of such structures; for example, R and S configurations at various asymmetric centers, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers. Therefore, single stereochemical isomers as well as enantiomeric mixtures, diastereomeric mixtures, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention.

[0134] Any asymmetric atom (e.g., carbon, etc.) of the compounds of the invention may exist in a racemic or enantiomerically enriched form, such as in the (R)-, (S)-, or (R, S)-configuration. In certain embodiments, each asymmetric atom has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess, or at least 99% enantiomeric excess in terms of (R)- or (S)-configuration. Substituents on atoms with unsaturated double bonds may exist in cis-(Z)- or trans-(E)-forms, if possible.

[0135] Therefore, as described herein, the compounds of the present invention may exist in the form of one or a mixture of possible isomers, rotamers, atropisomers, tautomers, for example, in the form of substantially pure geometric (cis or trans) isomers, diastereomers, optical isomers (enantiomers), racemates or mixtures thereof.

[0136] Any resulting mixtures of isomers can be separated on the basis of the physicochemical differences of the components into the pure or substantially pure geometric or optical isomers, diastereomers, racemates, for example, by chromatography and / or fractional crystallization.

[0137] Any racemate of the final product or intermediate obtained can be separated into optical antipodes by known methods by methods familiar to those skilled in the art, such as by separation of the salts of the diastereoisomers obtained. The racemic product can also be separated by chiral chromatography, such as high pressure liquid chromatography (HPLC) using a chiral adsorbent. In particular, enantiomers can be prepared by asymmetric synthesis (e.g., Jacques, et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Principles of Asymmetric Synthesis (2nd Ed. Robert E. Gawley, Jeffrey Aubé, Elsevier, Oxford, UK, 2012); Eliel, EL Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, SH Tables of Resolving Agents and Optical Resolutions p. 268 (EL Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN 1972).

[0138] As used herein, the term "tautomer" or "tautomeric form" refers to structural isomers with different energies that can be interconverted through a low energy barrier. If tautomerism is possible (such as in solution), a chemical equilibrium of tautomers can be reached. For example, proton tautomers (protontautomers) (also known as prototropic tautomers) include interconversions performed by proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions performed by the reorganization of some bonding electrons. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are within the scope of the present invention.

[0139] As used herein, the term "solvate" refers to an association formed by one or more solvent molecules and the compounds of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, aminoethanol. The term "hydrate" refers to an association formed by a solvent molecule that is water.

[0140] As used herein, the term "pharmaceutically acceptable" means that the substance or composition must be chemically and / or toxicologically compatible with the other ingredients comprising the formulation and / or the mammal to be treated therewith.

[0141] As used herein, the term "pharmaceutically acceptable salt" refers to organic and inorganic salts of the compounds of the present invention. Pharmaceutically acceptable salts are well known in the art, as described in the literature: SM Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66: 1-19. Pharmaceutically acceptable salts formed by non-toxic acids include, but are not limited to, inorganic acid salts (such as hydrochlorides, hydrobromides, phosphates, sulfates, perchlorates) formed by reaction with amino groups, and organic acid salts (such as acetates, oxalates, maleates, tartrates, citrates, succinates, malonates), or other methods described in books and literature, such as ion exchange methods, to obtain these salts. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentylpropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C 1～4 The present invention also contemplates quaternary ammonium salts formed by any compound containing a N group. Water-soluble or oil-soluble or dispersible products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, non-toxic ammonium, quaternary ammonium salts and amine cations formed by counter-balancing ions, such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, C 1～8 Sulfonates and aromatic sulfonates.

[0142] As used herein, the terms "optionally", "optional" or "optionally" generally mean that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs and instances where it does not.

[0143] In this article, the terms "optionally substituted", "optionally substituted" and "substituted or unsubstituted" can be used interchangeably. In general, the term "optionally", whether or not it is preceded by the term "substituted", means that one or more hydrogen atoms in a given structure are replaced by a specific substituent. Unless otherwise indicated, an optional substituent group can be substituted at each substitutable position of the group. When more than one position in the given structural formula can be substituted by one or more substituents selected from a specific group, the substituents can be substituted at each position in the same or different manner. The substituents described can be, but are not limited to, F, Cl, Br, CN, OH, NH2, NO2, etc.

[0144] As used herein, the term "one or more" (e.g. in the definition of substituents of compounds of the general formula of the invention) means "one, two, three, four or five, especially one, two, three or four, more especially one, two or three, even more especially one or two".

[0145] In addition, it should be noted that, unless explicitly stated otherwise, the description methods used in the present invention, "each... is independently" and "... are each independently" and "... are independently" can be interchanged and should be understood in a broad sense, which can mean that in different groups, the specific options expressed by the same symbols do not affect each other, or that in the same group, the specific options expressed by the same symbols do not affect each other.

[0146] As used herein, the term "halogen" refers to a fluorine, chlorine, bromine or iodine atom.

[0147] In this document, the minimum and maximum carbon atom content in a hydrocarbon group is indicated by a prefix, for example, the prefix C a～b It means containing "a" to "b" carbon atoms. 1～n ” refers to a straight or branched saturated / unsaturated carbon chain containing 1, 2, 3, 4, 5, ... or n carbon atoms; further understand that “C 1～n " shall be interpreted as including any sub-ranges included therein, such as C 1～6 Including C 1～5 , C 1～4 , C 1～3 , C 1～2 , C 2～5 , C 2～4 , C 2～3 , C3～5 , C 3～4 , C 4～5 .

[0148] In this article, the term “C 1～6 "Alkyl" refers to a straight-chain or branched saturated monovalent hydrocarbon group having 1, 2, 3, 4, 5 or 6 carbon atoms, such as C 1～5 Alkyl, C 1～4 Alkyl, C 1～3 Alkyl, C 2～5 Alkyl, C 2～4 Alkyl, C 2～3 Alkyl. These include, but are not limited to, methyl, ethyl, n-propyl (n-Pr, -CH2CH2CH3), isopropyl (i-Pr, -CH(CH3)2), n-butyl (n-Bu, -CH2CH2CH2CH3), isobutyl (i-Bu, -CH2CH(CH3)2), sec-butyl (s-Bu, -CH(CH3)CH2CH3), tert-butyl (t-Bu, -C(CH3)3), n-pentyl (-CH2CH2CH2C H2CH3), 2-pentyl (-CH(CH3)CH2CH2CH3), 3-pentyl (-CH(CH2CH3)2), 2-methyl-2-butyl (-C(CH3)2CH2CH3), 3-methyl-2-butyl (-CH(CH3)CH(CH3)2), 3-methyl-1-butyl (-CH2CH2CH(CH3)2), 2-methyl-1-butyl (-CH2CH(CH3)CH2CH3), n-hexyl ( -CH2CH2CH2CH2CH2CH3), 2-hexyl (-CH(CH3)CH2CH2CH2CH3), 3-hexyl (-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl (-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl (-CH(CH3)CH(CH3)CH2CH3), 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3), wherein the alkyl groups may independently be unsubstituted or substituted with one or more substituents described herein.

[0149] In this article, the term “C 2～6"Alkenyl" refers to a linear or branched monovalent hydrocarbon group having 2, 3, 4, 5 or 6 carbon atoms, wherein at least one position CC is an sp2 double bond unsaturated state, wherein the alkenyl group can be independently unsubstituted or substituted by one or more substituents described in the present invention, including groups with "cis", "trans" or "Z" "E" orientation, wherein specific examples include but are not limited to ethyl (-CH=CH2), allyl (-CH2CH=CH2), etc. For example, C 2～5 Alkenyl, C 2～4 Alkenyl, C 2～3 Alkenyl, C 3～6 Alkenyl, C 3～5 Alkenyl, C 3～4 Alkenyl.

[0150] In this article, the term “C 2～6 "Alkynyl" refers to a linear or branched monovalent hydrocarbon group having 2, 3, 4, 5 or 6 carbon atoms, wherein at least one position CC is an unsaturated sp triple bond state, wherein the alkynyl group may be independently unsubstituted or substituted with one or more substituents described in the present invention, specific examples include but are not limited to alkynylethyl (-C≡CH2), propargyl (-CH2C≡CH), 1-propynyl (-C≡C-CH3) and the like. For example, C 2～5 Alkynyl, C 2～4 Alkynyl, C 2～3 Alkynyl, C 3～6 Alkynyl, C 3～5 Alkynyl, C 3～4 Alkynyl.

[0151] In this article, the term “C 1～6 "Alkoxy" refers to a C 1～6 Alkyl, wherein the term "alkyl" is as defined above. For example: methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, pentyloxy, isopentyloxy and n-hexyloxy, or isomers of the above groups. In particular, the "C1-C6 alkoxy" may contain 1, 2, 3, 4 or 5 carbon atoms ("C 1～5 Alkoxy”), preferably, may contain 1, 2, 3 or 4 carbon atoms (“C 1～4 "alkoxy").

[0152] In this article, the term “C 1～6 "Cycloalkyl" or "1 to 6-membered cycloalkyl" refers to a saturated monovalent mono- or bicyclic hydrocarbon ring containing 1, 2, 3, 4, 5 or 6 carbon atoms. 3～6 Cycloalkyl is a monocyclic hydrocarbon ring, for example, cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl.

[0153] In this context, the term "alkylene" refers to a group formed by removing a hydrogen atom from an "alkyl" group, wherein "C 1～6 "Alkylene" includes methylene, ethylene, propylene, isopropylene (such as ), butylene (such as ), pentylene (such as ), hexamethylene (such as )wait.

[0154] As used herein, the term "cycloalkylene" refers to a group formed by removing a hydrogen atom from a "cycloalkyl".

[0155] In this document, the terms "heterocycloalkyl", "heterocycle" and "heterocycloalkane" all refer to a saturated ring or a non-aromatic unsaturated ring containing at least one heteroatom; wherein the heteroatom refers to a nitrogen atom, an oxygen atom, a sulfur atom, etc. It usually refers to a monovalent saturated or partially unsaturated monocyclic or bicyclic ring system of multiple ring atoms, which contains 1, 2 or 3 ring heteroatoms selected from N, O and S, and the remaining ring atoms are carbon.

[0156] The 4-7 membered heterocycle described in the present invention refers to a 4-, 5-, 6- or 7-membered saturated or unsaturated heterocycle, wherein the unsaturated refers to a group or molecule containing a carbon-carbon double bond, a carbon-carbon triple bond, a carbon-oxygen double bond, a carbon-sulfur double bond, a carbon-nitrogen triple bond, etc.

[0157] As used herein, the terms "-OR", "-NRR" and the like refer to an R group being connected to an oxygen atom or a nitrogen atom by a single bond.

[0158] As used herein, the oxygen atom in the term "-C(O)R" or the like is connected to a carbon atom or a sulfur atom via a double bond.

[0159] It should be noted that the term "C 1～6 ”, for example, in “C 1～6 Alkyl" or "C 1～6 In the context of the definition of "alkoxy", it refers to an alkyl group having a limited number of carbon atoms from 1 to 6, i.e., 1, 2, 3, 4, 5 or 6 carbon atoms. It is further understood that the term "C 1～6 " shall be interpreted as including any sub-ranges included therein, such as C 1～6 , C 2～5 , C 3～4 , C 1～2 , C 1～3 , C 1～4 , C 1～5 ; especially C 1～2 , C 1～3 , C 1～4 , C 1～5 , C 1～6 ; especially C 1～4 .

[0160] Similarly, as used herein, the term “C 2～6 ”, for example, in “C 2～6 "Alkenyl" and "C 2～6 In the context of the definition of "alkynyl", it is understood to refer to alkenyl or alkynyl groups having a limited number of carbon atoms of 2 to 6, i.e. 2, 3, 4, 5 or 6 carbon atoms. It is further understood that the term "C 2～6 " shall be interpreted as including any sub-ranges therein, e.g., C 2～6 , C 3～5 , C 3～4 , C 2～3 , C 2～4 , C 2～5 ; especially C 2～3 .

[0161] The group description of the present invention It is used to describe the position of group substitution.

[0162] In this article, the term "pharmaceutically acceptable excipients" includes any solvent, dispersion medium, coating material, surfactant, antioxidant, preservative (e.g., antibacterial agent, antifungal agent), isotonic agent, salt, drug stabilizer, adhesive, excipient, dispersant, lubricant, sweetener, flavoring agent, coloring agent, or a combination thereof, which are all known to those skilled in the art (such as Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329). Except for the case where any conventional carrier is incompatible with the active ingredient, its use in treatment or pharmaceutical composition is covered.

[0163] As used herein, the term "administration" refers to the introduction of a predetermined amount of a substance into a patient by some suitable means. The recombinant cell or pharmaceutical composition of the present invention can be administered by any common route as long as it can reach the desired tissue. Various modes of administration are contemplated, including peritoneal, intravenous, intramuscular, subcutaneous, etc., but the present invention is not limited to these exemplified modes of administration. Preferably, the composition of the present invention is administered by intravenous or subcutaneous injection.

[0164] As used herein, the term "treatment" refers to the use of drugs to obtain the desired pharmacological and / or physiological effects. The effect may be preventive in terms of completely or partially preventing a disease or its symptoms, and / or may be therapeutic in terms of partially or completely curing a disease and / or the adverse effects caused by the disease. "Treatment" as used herein covers diseases in mammals, particularly humans, and includes: (a) preventing the occurrence of a disease or condition in an individual who is susceptible to the disease but has not yet been diagnosed with the disease; (b) inhibiting the disease, such as blocking the progression of the disease; or (c) alleviating the disease, such as alleviating symptoms associated with the disease. "Treatment" as used herein covers any medication that administers a drug or compound to an individual to treat, cure, alleviate, improve, reduce or inhibit an individual's disease, including but not limited to administering a drug containing a compound described herein to an individual in need.

[0165] In this article, the term "cancer" or "tumor" can be any unregulated cell growth. Exemplarily, it can be lung cancer (e.g., non-small cell lung cancer), papillary thyroid cancer, glioblastoma multiforme, colon cancer, rectal cancer, head and neck cancer, kidney cancer, bladder cancer, breast cancer, ovarian cancer, liver cancer, bile duct cancer, sarcoma, acute myeloid leukemia, large cell neuroendocrine cancer, neuroblastoma, prostate cancer, neuroblastoma, pancreatic cancer, melanoma, head and neck squamous cell carcinoma, cervical cancer, skin cancer, glioma, esophageal cancer, oral squamous cell carcinoma or gastric cancer, etc.

[0166] The scheme of the present invention will be explained below in conjunction with the embodiments. It will be appreciated by those skilled in the art that the following embodiments are only used to illustrate the present invention and should not be considered as limiting the scope of the present invention. Where specific techniques or conditions are not indicated in the embodiments, the techniques or conditions described in the literature in this area or the product specifications are used. The reagents or instruments used are not indicated by the manufacturer and are all conventional products that can be obtained commercially.

[0167] Example 1: Preparation of Monomer Compounds

[0168] 1. Synthesis of compounds M-1 and M-2

[0169]

[0170] Step 1: 4-dimethylaminopyridine (4.5 mg, 0.037 mmol), di-tert-butyl dicarbonate (122.2 mg, 0.56 mmol) and triethylamine (154.3 uL, 1.11 mmol) were added to a dichloromethane (10 mL) solution of 5,6-dimethoxy-1H-indole (65.5 mg, 0.37 mmol). The reaction solution was stirred at room temperature for 4 hours, 10 mL of water was added to quench the reaction, and it was extracted with dichloromethane. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column to obtain M-1-int-1 (94.2 mg, 0.34 mmol, 93% yield).

[0171] Step 2: Under nitrogen protection, 2,2,6,6-tetramethylpiperidine (TEMP, 72 mg, 0.51 mmol) was dissolved in 1 mL of anhydrous tetrahydrofuran. At -78°C, n-butyl lithium (20.4 uL, 0.51 mmol, 2.5 M in n-hexane) was added dropwise to the above reaction flask and stirred at this temperature for 1 hour. A tetrahydrofuran solution (2 mL) of M-1-int-1 (94.2 mg, 0.34 mmol) was added dropwise to the above reaction solution. After the reaction solution was stirred at -78°C for one hour, a tetrahydrofuran solution (1 mL) of 2,2-dimethylsuccinic anhydride (87.1 mg, 0.68 mmol) was added to the above reaction solution, and the mixture was stirred at -78°C for 1 hour, warmed to room temperature and stirred overnight. After the reaction was completed, a small amount of water and hydrochloric acid were added to the reaction solution to quench the reaction, and the pH was adjusted to 3. The crude product was concentrated and purified using a silica gel column (20:80 = ethyl acetate: petroleum ether) to give a mixture of M-1-int-2 and M-1-int-3 (48.7 mg, 0.12 mmol, 35% yield).

[0172] Step 3: Trifluoroacetic acid (89.1uL, 1.2mmol) was added dropwise to a dichloromethane (2mL) solution of M-1-int-2 and M-1-int-3 (48.7mg, 0.12mmol) and stirred at room temperature for 2 hours. After the reaction was completed, ethyl acetate and water were used for extraction, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by silica gel column (20:80 = ethyl acetate: petroleum ether) to obtain a mixture of M-1-int-4 and M-1-int-5 (33.6mg, 0.11mmol, 96% yield).

[0173] Step 4: Diazomethane (165uL, 0.33mmol, 2.0M hexane solution) was added dropwise to a solution of M-1-int-4 and M-1-int-5 (33.6mg, 0.11mmol) in methanol (1mL) and ether (1mL), and the mixture was reacted at room temperature for 2h. After the reaction was completed, the reaction solution was concentrated, the residue was extracted with ethyl acetate and water, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and finally purified using a silica gel column to obtain a mixture of M-1-int-6 and M-1-int-7 (31.9mg, 0.10mmol, 91% yield).

[0174] Step 5: Cesium carbonate (48.9 mg, 0.15 mmol) and iodomethane (18.7 uL, 0.3 mmol) were added to a mixture of M-1-int-6 and M-1-int-7 (31.9 mg, 0.10 mmol) in acetonitrile (2 mL), the temperature was raised to 80 ° C, and the reaction was carried out for 12 hours. After the reaction was completed, the reaction solution was concentrated, the residue was extracted with ethyl acetate and water, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified by preparative chromatography to obtain M-1 (9.3 mg, 0.028 mmol, 28% yield) and M-2 (17 mg, 0.051 mmol, 51% yield), respectively.

[0175] M-1:Purity:98%; HRMS(ESI)m / z:[M+H] + Calcd for C 18 H 24 NO5 334.1654; Found334.1658; 1 H NMR (600MHz, DMSO-d6) δ7.25(s,1H),7.09(s,1H),7.03(s,1H),3.85(s,3H),3.81(s,3H),3.76(s,3H),3.49(s,3H),2.87(s,2H),1.41(s,6H).

[0176] M-2:Purity:97%; HRMS(ESI)m / z:[M+H] + Calcd for C 18 H 24 NO5 334.1654; Found334.1654; 1 H NMR (600MHz, DMSO-d6) δ7.36(s,1H),7.10(s,1H),7.04(s,1H),3.93(s,3H),3.86(s,3H),3.77(s,3H),3.57(s,3H),3.26(s,2H),1.22(s,6H).

[0177] 2. Synthesis of M-3 compound

[0178]

[0179] 33 mg of M-1 (0.1 mmol) prepared in Part 1 of this Example was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 12 mg LiOH (0.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography to obtain 16 mg M-3.

[0180] M-3:Purity:99%; HRMS(ESI)m / z:[MH] - Calcd for C 17 H 20 NO5 318.1341; Found318.1360; 1 H NMR (600MHz, DMSO-d6) δ7.34(s,1H),7.10(s,1H),7.04(s,1H),3.94(s,3H),3.86(s,3H),3.77(s,3H),3.19(s,2H),1.20(s,6H).

[0181] 3. Synthesis of M-4 compound

[0182]

[0183] 33 mg of M-2 (0.1 mmol) prepared in Part 1 of this Example was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 12 mg LiOH (0.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography to obtain 13 mg of M-4.

[0184] M-4:Purity:99%; HRMS(ESI)m / z:[MH] - Calcd for C 17 H 20 NO5 318.1341; Found318.1354; 1H NMR (600MHz, DMSO-d6) δ7.17(s,1H),7.09(s,1H),7.02(s,1H),3.85(s,3H),3.78(s,3H),3.76(s,3H),2.79(s,2H),1.36(s,6H).

[0185] 4. Synthesis of compounds M-5 and M-6

[0186]

[0187] At -78 ° C, t-BuLi (0.94 mg, 0.72 mmol) was added dropwise to a THF (2.0 mL) solution containing M-1-int-1 (100.0 mg, 0.36 mmol) prepared in Part 1 of this Example, and then the mixture was heated to room temperature and stirred for 16 hours. The reaction was quenched with saturated aqueous ammonium chloride (10.0 mL) at 0 ° C, diluted with water (5.0 mL), extracted with EtOAc (10.0 mL), and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by preparative thin layer (PE: EA = 5: 1) to obtain white solids M-5 (25.0 mg, 0.063 mmol, yield 17.7%) and M-6 (10 mg, 0.025 mmol, yield 7.1%).

[0188] M-5:MS m / z(ESI):336.0[M+H] + ; 1 H NMR (400MHz, DMSO) δ12.23(s,1H),7.47(s,1H),7.42(s,1H),7.21(s,1H),3.84(s,3H),3.81(s,3H),3.26(dd,J =17.2,7.4Hz,1H),2.96(dd,J=17.2,5.6Hz,1H),2.86(dd,J=13.4,7.2Hz,1H),1.54(s,9H),1.33-1.10(m,3H).

[0189] M-6:MS m / z(ESI):336.1[M+H] + ; 1H NMR (400MHz, CDCl3) δ7.62(s,1H),7.20(s,1H),7.01(s,1H),3.98(s,3H),3.93(s,3H),3.67(dd,J=13.4 ,7.2Hz,1H),2.97(dd,J=16.8,7.6Hz,1H),2.50(dd,J=16.8,5.6Hz,1H),1.59(s,9H),1.36-1.22(m,3H).

[0190] 5. Synthesis of M-7 compound

[0191]

[0192] At -78 ° C, n-BuLi (0.56 mL, 0.56 mmol) was added dropwise to a THF (1.0 mL) solution containing 2,2,6,6-tetramethylpiperidine (TEMP, 59.0 mg, 0.41 mmol), and M-1-int-1 (50.0 mg, 0.18 mmol) prepared in Part 1 of this Example was added 15 min later, and the reaction was continued at -78 ° C for 40 min, 3,4-dimethylfuran-2,5-dione (45.0 mg, 0.36 mmol) was added, and the reaction was continued for 20 min. After the reaction was completed, the reaction was quenched with saturated NH4Cl (10.0 mL), diluted with water (5.0 mL), and extracted with EtOAc (10.0 mL), and the organic phase was dried over anhydrous Na2SO4, filtered, and concentrated. A thin layer was prepared (10-100% EtOAc / hexanes) to afford M-7 as a white solid (12.0 mg, 0.031 mmol, 16.6% yield).

[0193] M-1:MS m / z(ESI):348.1[M-56+H] + ; 1 H NMR(400MHz, CDCl3)δ8.62(s,1H),7.57(s,1H),6.91(s,1H),6.29(s,1H),3.93 (s,3H),3.90(s,3H),2.23(d,J=1.2Hz,3H),1.93(d,J=1.2Hz,3H),1.78(s,9H).

[0194] 6. Synthesis of compounds M-8, M-9 and M-10

[0195]

[0196] Step 1: n-BuLi (1.1 mL, 2.7 mmol) was added dropwise to a THF (5.0 mL) solution containing 2,2,6,6-tetramethylpiperidine (TEMP, 2.16 mmol) at -78°C, and M-1-int-1 (500 mg, 1.8 mmol) prepared in Part 1 of this Example was added after 15 min, and stirred for 40 min. 3-Oxabicyclo[3.1.0]hexane-2,4-dione (303.0 mg, 2.70 mmol) was added at -78°C, and the reaction was continued for 20 min. After completion of the reaction, the reaction was quenched with saturated aqueous NH4Cl solution (10.0 mL), diluted with water (5.0 mL), extracted with EtOAc (10.0 ml), and the organic phase was dried over anhydrous NaSO4, filtered, concentrated, and a thin layer was prepared (10-100% ethyl acetate / hexane) to give a white solid M-8-int-1 (100.0 mg, 0.25 mmol, yield 14.2%).

[0197] M-8-int-1:MS m / z(ESI):334.0[M-56+H] + .

[0198] Step 2: At 0°C, to a solution of M-8-int-1 (100.0 mg, 0.26 mmol) in EA (2.0 mL) was added aqueous HCl (92.5 mg, 2.57 mmol), stirred at room temperature for 4 h, concentrated, and the crude product was purified by column chromatography (DCM:MeOH=20:1) to give a brown liquid M-8 (80.0 mg, crude product).

[0199] Step 3: M-8 (80.0 mg, 0.28 mmol) was dissolved in DMF (2.0 mL), and NaOH (33.0 mg, 0.83 mmol) and CH3I (196.4 mg, 1.38 mmol) were added in sequence, and stirred at 45 °C for 16 h. After the reaction was completed, HCl aqueous solution (10.0 mL, 1 M) was added under ice bath, and extracted with EtOAc (10.0 mL). The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated. Preparative thin layer (10–100% ethyl acetate / hexane) was used for purification to obtain yellow solid M-9 (1.22 mg, 0.0038 mmol, yield 1.5%).

[0200] M-9:MS m / z(ESI):318.2[M+H] + ; 1H NMR (400MHz, CDCl3) δ7.37(s,1H),7.05(s,1H),6.73(s,1H),4.03(s,3H),3.99(s,3H),3.93(s ,3H),3.74(s,3H),3.14-3.10(m,1H),2.35-2.31(m,1H),1.64-1.51(m,1H),1.30-1.22(m,1H).

[0201] Step 4: M-9 (180.0 mg, crude) and LiOH (71.0 mg, 1.70 mmol) were added to THF / H2O (2.0 mL) at room temperature and stirred for 16 h. After the reaction was completed, HCl (10.0 mL, 1.0 M) was added under ice bath to quench the reaction, and EtOAc (10.0 mL×2) was used for extraction. The organic phase was dried over anhydrous Na2SO4, filtered, and concentrated. Preparative thin layer (10-100% DCM / MeOH) was used for purification to obtain a white solid M-10 (2.14 mg, 0.0038 mmol, yield 1.24%).

[0202] M-10:MS m / z(ESI):304.1[M+H] + ; 1 H NMR(400MHz,MeOD)δ7.48(s,1H),7.16(s,1H),6.94(s,1H),4.00(s,3H),3.94(s,3H),3 .86(s,3H),3.31-3.29(m,3H),3.16-3.12(m,1H),2.18-2.13(m,1H),1.53-1.44(m,2H).

[0203] 7. Synthesis of M-11 compound

[0204]

[0205] Step 1: n-BuLi (2.5M / hexane, 1.08 mL) was added dropwise to a solution of 2,2,6,6-tetramethylpiperidine (TEMP, 2.16 mmol) in THF (10.0 mL) at -78°C and stirred at constant temperature for 1 hour. Then M-1-int-1 (500 mg, 1.80 mmol) prepared in Part 1 of this Example and 3-oxabicyclo[3.2.0]heptane-2,4-dione (341.07 mg, 2.70 mmol) were added and stirred at -78°C for 1 hour. The reaction was quenched with ice-cold saturated aqueous ammonium chloride solution (20.0 mL), extracted with EtOAc (20.0 mL×2), washed with saturated brine (10.0 mL), dried over anhydrous Na2SO4, filtered and concentrated, and the residue was purified by flash column chromatography (PE:EA=2:1) ​​to give M-11-int-1 (108.0 mg, 267.70 umol, yield 14.9%).

[0206] M-11-int-1:MS m / z(ESI):348.1[M-56+H] + .

[0207] Step 2: M-11-int-1 (80 mg, 198.30 umol), CH3I (42.2 mg, 297.45 umol) and K2CO3 (32.8 mg, 238.00 umol) were added to DMF (2.0 mL) in sequence and stirred at room temperature for 16 h. Water (20.0 mL) was added to quench the reaction, extracted with EtOAc (20.0 mL × 2), washed with saturated brine (15.0 mL), dried over anhydrous Na2SO4, filtered and concentrated, and the residue was purified by preparative thin layer (petroleum ether: EtOAc = 5:1) to obtain colorless oily 5,6-dimethoxy-2-(2-methoxycarbonylcyclobutanecarbonyl) indole-1-carboxylic acid tert-butyl ester M-11-int-2 (28.0 mg, 67.07 umol, yield 33.8%).

[0208] M-11-int-2:MS m / z(ESI):362.1[M-56+H] + .

[0209] Step 3: M-11-int-2 (62.0 mg, 148.52 umol) and TFA (16.93 mg, 148.5 2 umol) were dissolved in DCM (3 mL) and stirred at room temperature for 3 hours. Concentrated, diluted with water, extracted with EtOAc (20.0 mL × 2), washed with saturated brine (15.0 mL × 3), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by TLC (petroleum ether: EtOAc = 2: 1) to obtain 5-(5,6-dimethoxy-1-methylindol-2-yl)-5-oxo-pentanoic acid methyl ester M-11-int-3 (30.0 mg, 93.94 umol, yield 24.88%) as a colorless oil.

[0210] M-11-int-3:MS m / z(ESI):318.2[M+H] + .

[0211] Step 4: M-11-int-3 (27 mg, 85.08 umol) was dissolved in DMF (2.0 mL), sodium hydride (2.45 mg, 102.10 umol) was added in batches at 0 ° C, stirred at constant temperature for 1 hour, CH3I (24.15 mg, 170.17 umol) was added, and the mixture was heated to room temperature and stirred for 16 hours. The reaction was quenched with ice water, extracted with EtOAc (2x20 mL), washed with saturated brine (15.0 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative thin layer (PE: EtOAc = 2: 1) to obtain colorless oily 2-(5,6-dimethoxy-1-methylindole-2-carbonyl) cyclobutanecarboxylic acid methyl ester M-11-int-4 (24.0 mg, 72.43 umol, yield 85.13%).

[0212] M-11-int-4:MS m / z(ESI):332.2[M+H] + ;

[0213] Step 5: M-11-int-4 (20.0 mg, 60.36 umol) and LiOH (1.7 mg, 72.43 umol) were dissolved in THF:H2O=5:1 (6.0 mL) and stirred at room temperature for 3 hours. Water was added for dilution, and EtOAc (2x 20.0 mL) was extracted, washed with brine (15.0 mL), dried over anhydrous Na2SO4, filtered and concentrated. The residue was purified by preparative thin layer (petroleum ether: EtOAc=2:1) ​​to obtain a white solid 2-(5,6-dimethoxy-1-methylindole-2-carbonyl)cyclobutanecarboxylic acid M-11 (6.0 mg, 18.91 umol, yield 31.33%).

[0214] M-11:MS m / z(ESI):318.1[M+H] + ; 1 H NMR (400MHz, MeOD) δ7.34(s,1H),7.12(s,1H),6.94(s,1H),4.31-4.23(m,1H),4.03(s,3H),3.93(s,3H),3.85(s,3H),2.26-2.15(m,4H).

[0215] 8. Synthesis of M-12 compound

[0216]

[0217] Step 1: M-1-int-1 (300 mg, 1.08 mmol) prepared in Part 1 of this Example was dissolved in THF (2.0 mL), t-Buli (1.6 M / hexane, 1.35 mL) was slowly added dropwise at -78 ° C, stirred at constant temperature for 40 min, hexahydroisobenzofuran-1,3-dione (250.16 mg, 1.62 mmol) was added, and the mixture was heated to room temperature and stirred overnight. The reaction was quenched with saturated NH4Cl aqueous solution (10.0 mL), diluted with water (5.0 mL), extracted with EtOAc (10.0 ml), dried over anhydrous Na2SO4, filtered, and concentrated. Silica gel chromatography (10–100% EtOAc / hexane, linear gradient; diatomaceous earth dry loading) gave a white solid M-12-int-1 (220 mg, 509.88 umol, yield 47.13%).

[0218] M-12-int-1:MS m / z(ESI):376.00[M-56+H] + .

[0219] Step 2: M-12-int-1 (170.0 mg, 393.99 umol) was dissolved in HCl / EA (2.0 mL) and stirred at room temperature for 16 hours. The residue was concentrated and purified by column chromatography (DCM: MeOH = 20: 1) to obtain M-12-int-2 (120.0 mg, 362.14 umol, yield 91.92%) as a brown oil.

[0220] M-12-int-2:MS m / z(ESI):332.05[M+H] + .

[0221] Step 3: M-12-int-2 (80.0 mg, 241.43 umol), NaOH (29.0 mg, 0.83 mmol) and MeI (171.34 mg, 1.21 mmol) were added to DMF (2.0 mL) and stirred at 45 ° C for 16 hours. HCl (10.0 mL, 1 M) was added to quench the reaction, extracted with EtOAc (10.0 mL), dried over anhydrous Na2SO4, filtered, concentrated, and column chromatography (10-100% EtOAc / hexane) gave a white solid M-12 (7.4 mg, 21.45 umol, 8.88% yield).

[0222] M-12:MS m / z(ESI):346.0[M+H] + ; 1 H NMR (400MHz, Methanol-d4) δ7.24(s,1H),7.14(s,1H),6.94(s,1H),3.87(d,J=10.2Hz,6H),3.51(s,3H),3.50 -3.42(m,1H),2.88-2.76(m,1H),2.17-2.06(m,2H),1.92-1.81(m,2H),1.58-1.45(m,2H),1.45-1.32(m,2H).

[0223] 9. Synthesis of M-13 and M-14 compounds

[0224]

[0225] Step 1: Add TEMP (2.16 mmol) to tetrahydrofuran (6.0 mL) under stirring at -78°C, then add n-butyl lithium (2.70 mmol), and stir the mixture for 1 hour. M-1-int-1 (500.0 mg, 1.80 mmol) and glutaric anhydride (308.6 mg, 2.70 mmol) prepared in Part 1 of this Example are added to the above mixture in sequence, and the reaction is stirred at -78°C for 1 hour. The resulting mixture is diluted with ice water and extracted with ethyl acetate (2 x 20.0 mL), washed with saturated brine (10.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue is purified by silica gel column (PE: EA = 2: 1) to obtain brown oily liquid M-13-int-1 (230.0 mg, 587.61 umol, yield 32.67%).

[0226] M-13-int-1:MS m / z(ESI):292.2[M-100+H] + .

[0227] Step 2: M-13-int-1 (230.0 mg, 587.61 umol), trifluoroacetic acid (335.0 mg, 2.94 mmol) and dichloromethane (6.0 mL) were mixed and stirred at room temperature for 3 hours. The resulting mixture was diluted with water and extracted with ethyl acetate (2x20.0 mL), washed with saturated brine (2x 15.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative thin layer (petroleum ether: ethyl acetate = 2: 1) to obtain a colorless oily liquid M-13-int-2 (130.0 mg, 446.28 umol, 75.95% yield).

[0228] M-13-int-2:MS m / z(ESI):292.1[M+H] + .

[0229] Step 3: M-13-int-2 (110.0 mg, 377.62 umol), iodomethane (214.4 mg, 1.51 mmol) and sodium hydroxide (30.2 mg, 755.24 umol) were added to DMF (4.0 mL), and the resulting mixture was stirred at 45 ° C for 16 hours. The resulting mixture was diluted with water and extracted with ethyl acetate (2 x 20.0 mL), washed with saturated brine (2 x 15.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative plate (petroleum ether: ethyl acetate = 2: 1) to obtain a colorless oily liquid M-13 (30.0 mg, 93.94 umol, 24.88% yield).

[0230] M-13:MS m / z(ESI):320.0[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.34(s,1H),7.11(s,1H),7.05(s,1H),3.98(s,3H),3.87(s,3H), 3.78(s,3H),3.60(s,3H),2.95(t,J=7.2Hz,2H),2.40(t,J=7.2Hz,2H),1.94-1.82(m,2H).

[0231] Step 4: A mixture of M-13 (30.0 mg, 93.94 umol) and lithium hydroxide (2.70 mg, 112.73 umol) was added to a mixture of tetrahydrofuran: water = 5:1 (6 mL) and stirred at room temperature for 2 hours. The resulting mixture was diluted with water, extracted with ethyl acetate (2x20 mL), washed with saturated brine (2x15 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified on a preparative plate (petroleum ether: ethyl acetate = 2:1) to obtain a white solid M-14 (6.0 mg, 19.65 umol, 20.92% yield).

[0232] M-14:MS m / z(ESI):305.95[M+H] + ; 1 H NMR(400MHz,MeOD)δ7.20(s,1H),7.12(s,1H),6.94(s,1H),3.88(s,3H),3.85(s,3H),3 .66(s,3H),2.96(t,J=7.2Hz,2H),2.44(t,J=7.2Hz,2H),2.03(dd,J=14.8,7.2Hz,2H).

[0233] 10. Synthesis of M-15 compound

[0234]

[0235] Step 1: 5-methoxy-4-azaindole (600.0 mg, 4.05 mmol), DMAP (494.7 mg, 4.05 mmol), triethylamine (818.03 mg, 8.08 mmol) and di-tert-butyl dicarbonate (1.33 g, 6.07 mmol) were added to a dichloromethane (6.0 mL) solution, and the resulting mixture was stirred at room temperature for 16 hours. The resulting mixture was concentrated under vacuum, diluted with water and extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column (PE: EA = 5: 1) to obtain a white solid M-15-int-1 (720.0 mg, 2.90 mmol, 71.61% yield).

[0236] M-15-int-1:MS m / z(ESI):249.0[M+H] + .

[0237] Step 2: At -78 ° C, tert-butyl lithium (1.61 mmol, 1.24 mL) was slowly added to the stirred mixture of M-15-int-1 (200.0 mg, 806.32 umol) and tetrahydrofuran (6.0 mL), and the mixture was stirred at -78 ° C for 1 hour. 3-methyltetrahydrofuran-2,5-dione (138.0 mg, 1.21 mmol) was added to the solution, and the reaction was stirred at room temperature for 16 hours. The resulting mixture was diluted with ice water and extracted with ethyl acetate (2x 20.0 mL), washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by preparative thin layer (petroleum ether: ethyl acetate = 2: 1) to obtain M-15-int-2 (12.7 mg, 0.035 mmol, yield 4.38%).

[0238] M-15-int-2:MS m / z(ESI):363.1[M+H] + .

[0239] Step 3: Ethyl acetate (3.0 mL) was added to a mixture of M-15-int-2 (12.7 mg, 0.035 mmol, 1.00 eq) and HCl / EtOAc (3 M, 1.0 mL) and stirred at room temperature for 2 hours. The resulting mixture was concentrated under vacuum, diluted with water, extracted with ethyl acetate (2 x 10.0 mL), washed with saturated brine (2 x 10.0 mL), dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified on a preparative plate (petroleum ether: ethyl acetate = 2: 1) to give M-15 (2.7 mg, 0.010 mmol, 29.4% yield) as a white solid.

[0240] M-15:MS m / z(ESI):263.1[M+H] + ; 1 H NMR (400MHz, MeOD) δ7.77 (d, J = 8.8Hz, 1H), 7.22 (s, 1H),

[0241] 6.78(d,J=8.8Hz,1H),3.95(s,3H),3.42(dd,J=18.4,9.6Hz,1H),3.05(d,J=12.8Hz,1H),1.28(d,J=6.8Hz,3H).

[0242] 11. Synthesis of compounds M-16, M-17, M-18 and M-19

[0243]

[0244] Step 1: At 0°C, sodium hydride (481.4 mg, 12.04 mmol, 60% dispersed in mineral oil) was added in batches to a DMF (20.0 mL) solution containing ethyl 5,6-dimethoxyindole-2-carboxylate (2.0 g, 8.02 mmol), and the resulting mixture was stirred at room temperature for 0.5 hours, then iodomethane (3.4 g, 24.07 mmol) was added dropwise, and then stirred for 3 hours. The reaction was quenched with saturated aqueous ammonium chloride (50.0 mL) at 0°C, extracted with ethyl acetate (20.0 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated to obtain M-16-int-1 (2.1 g, 7.98 mmol, yield 99.4%) as a white solid.

[0245] M-16-int-1:MS m / z(ESI):264.1[M+H] + .

[0246] Step 2: At -78 ° C and argon protection, bis(trimethylsilyl)amide lithium (1M, 8.36 mL, 8.36 mmol) was added dropwise to a THF (10 mL) solution containing dimethyl methylphosphonate (777.6 mg, 6.27 mmol), and the mixture was stirred at -78 ° C for 1 hour. Then a solution of M-16-int-1 (1.1 g, 4.18 mmol) in 5 ml THF was added dropwise to the mixture. The mixture was heated to room temperature and stirred for 2 hours. The reaction was quenched with saturated aqueous ammonium chloride solution (100.0 mL). DCM (50.0 mL × 3) was used for extraction, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was purified by gel column chromatography (DCM: MeOH = 20: 1) to give a white solid M-16-int-2 (1.2 g, 3.52 mmol, yield 84.2%).

[0247] M-16-int-2:MS m / z(ESI):342.1[M+H] + .

[0248] Step 3: At 0 ° C and argon protection, sodium hydride (220.0 mg, 5.50 mmol, 60% dispersed in mineral oil) was added to a THF (10.0 mL) solution containing M-16-int-2 (1.25 g, 3.67 mmol) in batches, and the resulting mixture was stirred at 0 ° C for 30 minutes. Then methyl 2-oxopropanoate (561.5 mg, 5.50 mmol) was added dropwise at 0 ° C for 10 minutes, and then stirred at room temperature for 2 hours. The reaction was quenched with ice water (100.0 mL) and extracted with ethyl acetate (50.0 mL × 2). The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by gel column chromatography (PE: EA = 8: 1) to obtain a bright yellow solid M-16-int-3 (1.1 g, 3.47 mmol, 94.5% yield).

[0249] M-16-int-3:MS m / z(ESI):318.1[M+H] + .

[0250] Step 4: Pd / C (10%, 120.0 mg) was added to a MeOH (10.0 mL) solution containing SG-001-14 (1.18 g, 3.72 mmol). The resulting mixture was then stirred at room temperature under hydrogen for 3 hours. The mixture was filtered, the filtrate was concentrated and purified by gel column chromatography (PE: EA = 10: 1) to give a white solid M-16 (1.05 g, 3.29 mmol, 88.4% yield).

[0251] M-16:MS m / z(ESI):320.2[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.38(s,1H),7.11(s,1H),7.05(s,1H),3.95(s,3H),3.87(s,3H),3.78(s, 3H), 3.60 (s, 3H), 3.29-3.35 (m, 1H), 3.06-3.11 (m, 1H), 2.93-2.97 (m, 1H), 1.17 (d, J = 7.2Hz, 3H).

[0252] Step 5: Lithium hydroxide monohydrate (31.6 mg, 0.752 mmol) was added to a solution of MeOH (2.0 mL) and water (1.0 mL) containing M-16 (60.0 mg, 0.188 mmol) and stirred at room temperature for 16 hours. The mixture was adjusted to pH 3 with dilute hydrochloric acid (50.0 mL) and purified by Perp-HPLC (column: Waters Xbridge 20*150mm 10um 10um, C18, mobile phase A: water, mobile phase B: ACN) to give a white solid M-17 (30.0 mg, 0.10 mmol, yield 52.3%)

[0253] M-17:MS m / z(ESI):306.1[M+H] + ; 1 H NMR (400MHz, CDCl3) δ7.24(s,1H),7.03(s,1H),6.73(s,1H),4.04(s,3H),3.99( s,3H),3.93(s,3H),3.37-3.41(m,1H),3.07-3.17(m,2H),1.32(d,J=7.2Hz,3H).

[0254] Step 6: M-17 (45.0 mg, 0.147 mmol) was separated by chiral resolution (CO2 / MeOH / DEA 5%~40% 1.5 ml / min OJ, 3um, 3.0*100 (Daicel)); retention time / min, P1: 4.066 min, P2: 4.381 min) to give M-18 (15.2 mg, 0.0498 mmol, yield 33.9%, 99.42% ee) and M-19 (16.2 mg, 0.0531 mmol, yield 36.1%, 98.10% ee).

[0255] M-18 / M-19:MS m / z(ESI):306.1[M+H] + ;

[0256] M-18: 1 H NMR(400MHz,DMSO-d6)δ7.36(s,1H),7.11(s,1H),7.05(s,1H),3.96(s,3H),3.86 (s,3H),3.78(s,3H),3.30-3.33(m,1H),2.83-3.01(m,2H),1.14(d,J=7.2Hz,3H).

[0257] M-19: 1H NMR(400MHz,DMSO-d6)δ12.04(br.s,1H),7.37(s,1H),7.11(s,1H),7.05(s,1H),3.96(s,3 H),3.87(s,3H),3.78(s,3H),3.30-3.33(m,1H),2.86-3.11(m,2H),1.16(d,J=7.2Hz,3H).

[0258] 12. Synthesis of M-20 compound

[0259]

[0260] Methylamine hydrochloride (44.2 mg, 655.04 umol), HATU (64.7 mg, 170.31 umol) and N, N-diisopropylethylamine (169.3 mg, 1.31 mmol) were added to a solution of N, N-dimethylformamide (1.0 mL) containing M-17 (40.0 mg, 131.01 umol) prepared in Part 11 of this Example, and then stirred at room temperature for 16 hours. Water (10.0 mL) was added to the reaction solution and extracted with ethyl acetate (10.0 mL x 3). The organic phase was washed with water (10.0 mL x 3) and saturated brine (10.0 mL), dried over anhydrous sodium sulfate, concentrated, and purified on a silica gel column (petroleum ether: ethyl acetate = 1:1) to obtain a white solid M-20 (21.0 mg, 65.96 umol, yield 50.4%).

[0261] M-20:MS m / z(ESI):319.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.79 (d, J = 4.4Hz, 1H), 7.34 (s,

[0262] 1H),7.11(s,1H),7.04(s,1H),3.95(s,3H),3.87(s,3H),3.78(s,3H),3 .24(m,1H),2.92-2.77(m,2H),2.56(d,J=4.6Hz,3H),1.18-0.97(m,3H).

[0263] 13. Synthesis of M-21 compound

[0264]

[0265] HATU (59.8 mg, 0.16 mmol), N,N-diisopropylethylamine (50.8 mg, 0.39 mmol) and dimethylamine (0.197 mL, 0.39 mmol, 2M tetrahydrofuran solution) were added to a dichloromethane (2.0 mL) solution containing M-17 (40.0 mg, 0.13 mmol) prepared in Part 11 of this Example, and then stirred at room temperature for 16 hours. The reaction solution was concentrated and then purified by thin layer preparative chromatography (dichloromethane / methanol = 20 / 1) to obtain a white solid M-21 (8.3 mg, 25.10 umol, yield 19.2%).

[0266] M-21:MS m / z(ESI):333.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.36(s,1H),7.12(s,1H),7.04(s,1H),7.04(s,1H),3.94(s,3H),3.86(s,3H),3.78 (s,3H),3.42-3.33(m,2H),3.09(s,3H),2.89(dd,J=4.4Hz,16.8Hz,1H),2.81(s,3H),1.06(d,J=6.8Hz,3H).

[0267] 14. Synthesis of M-22 compound

[0268]

[0269] Step 1: M-17 (180.0 mg, 0.59 mmol) prepared in Part 11 of this Example was dissolved in DMF (3.0 mL), and O-benzylhydroxylamine (145.2 mg, 1.18 mmol), HATU (333.6 mg, 0.88 mmol) and N, N-diisopropylethylamine (380.9 mg, 2.95 mmol) were added. The reaction mixture was stirred at room temperature for 16 hours. The mixture was diluted with water (10.0 mL), extracted with ethyl acetate (20.0 mL×3), and washed with brine. It was dried over anhydrous sodium sulfate and concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate=1:1) to give a white solid compound M-22-int-1 (120.0 mg, 0.29 mmol, yield 62.2%).

[0270] M-22-int-1:MS m / z(ESI):411.3[M+H] + .

[0271] Step 2: M-22-int-1 (150.0 mg, 0.37 mmol) was dissolved in MeOH (3.0 mL) and Pd / C (15.0 mg, 10% wt, 50% water) was added, and the reaction mixture was stirred at room temperature for 6 hours under a hydrogen balloon. After the reaction was completed, the mixture was filtered and concentrated under reduced pressure, and purified by a reverse column (0.1% FA in water, acetonitrile) to obtain a white solid compound M-22 (6.44 mg, 0.020 mmol, 5.5% yield).

[0272] M-22:MS m / z(ESI):264.3[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.71(s,1H),7.03(s,1H),7.01(s,1H),6.90(s,1H),6.23(s,1H),3.88-3.83(m ,1H),3.82(s,3H),3.79(d,J=2.8Hz,1H),3.77(d,J=2.4Hz,1H),3.74(s,3H),3.67(s,3H),1.87(s,3H).

[0273] 15. Synthesis of M-23 compound

[0274]

[0275] Step 1: M-22-int-1 (50.0 mg, 0.12 mmol), copper acetate (22.1 mg, 0.12 mmol), 4-dimethylaminopyridine (44.7 mg, 0.37 mmol) and cyclopropylboronic acid (20.9 mg, 0.24 mmol) prepared in part 14 of this example were suspended in a toluene (1.0 mL) solution. NaHMDS (1 M tetrahydrofuran solution, 0.24 mL, 0.24 mmol) was added to the above solution. The reaction system was replaced with oxygen and stirred at 90 ° C for 16 hours. The reaction solution was filtered through diatomaceous earth and washed with dichloromethane, and the filtrate was concentrated and purified by high performance liquid preparative chromatography (mobile phase A: 0.1% trifluoroacetic acid aqueous solution, mobile phase B: acetonitrile) to obtain a white solid M-23-int-1 (8.0 mg, 17.76 umol, yield 14.6%).

[0276] M-23-int-1:MS m / z(ESI):451.3[M+H] + ; 1H NMR (400MHz, CDCl3) δ7.44-7.46(m,2H),7.32-7.39(m,4H),7.02(s,1H),6.72(s,1H),4.99-5.04(m,2H),4.01(s ,3H),3.98(s,3H),3.78(s,3H),3.48-3.60(m,2H),2.92-3.03(m,2H),1.18(d,J=6.8Hz,3H),0.88-0.96(m,4H).

[0277] Step 2: M-23-int-1 (20.0 mg, 44.39 umol) was dissolved in MeOH (2.0 mL) and Pd / C (4.0 mg, 20% wt.) was added. The reaction system was replaced with hydrogen and stirred at room temperature for 16 hours. The reaction solution was filtered through diatomaceous earth and the filtrate was concentrated. The residue was purified by high performance liquid preparative chromatography (mobile phase A: 0.1% trifluoroacetic acid aqueous solution, mobile phase B: acetonitrile) to obtain a white solid M-22 (6.0 mg, 16.65 umol, yield 37.5%).

[0278] M-23:MS m / z(ESI):361.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ9.57(br.s,1H),7.36(s,1H),7.12(s,1H),7.05(s,1H),3.95(s,3H),3.87(s,3H),3. 78(s,3H),3.37-3.50(m,1H),3.01-3.10(m,1H),2.85-2.90(m,1H),1.07(d,J=7.2Hz,3H),0.63-0.78(m,4H).

[0279] 16. Synthesis of M-24 and M-25 compounds

[0280]

[0281] Step 1: Add methoxyamine hydrochloride (164.1 mg, 1.97 mmol), HATU (298.9 mg, 786.05 umol) and N, N-diisopropylethylamine (846.6 mg, 6.55 mmol) to a solution of M-17 (0.2 g, 655.04 umol) prepared in Part 11 of this Example in N, N-dimethylformamide DMF (2.0 mL). The reaction solution was stirred at room temperature for 16 hours. The reaction solution was quenched with water (20.0 mL) and extracted with ethyl acetate (10.0 mL x 3). The organic phase was washed three times with water and then with saturated brine, dried over anhydrous sodium sulfate and concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1:1) to obtain an off-white solid M-24 (208.0 mg, 622.07 umol, yield 94.97%).

[0282] M-24:MS m / z(ESI):335.1[M+H] + .

[0283] Step 2: Sodium hydride (60%, 28.7 mg, 717.78 umol) was added to a solution of M-24 (0.2 g, 598.15 umol) in N, N-dimethylformamide (4.0 mL) at 0°C. The reaction solution was stirred at 0°C for 0.5 hours, and benzyl bromide (153.5 mg, 897.22 umol) was added dropwise to the reaction solution. The reaction solution was warmed to room temperature and stirred for 3 hours. The reaction solution was quenched with water (10.0 mL) and extracted with ethyl acetate (10.0 mL x 3), the organic phase was washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography to obtain a white solid M-25 (0.084 g, 204.65 umol, yield 34.21%).

[0284] M-25:MS m / z(ESI):425.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.38(s,1H),7.36-7.30(m,2H),7.27(d,J=6.0Hz,3H),7.12(s,1H),7.05(s,1H),4.89(d,J=16.0Hz, 1H),4.67(d,J=15.6Hz,1H),3.97(s,3H),3.87(s,3H),3.78(s,6H),3.43(m,2H),3.05-2.87(m,1H),1.13(t,J=14.4Hz,3H).

[0285] 17. Synthesis of compounds M-26, M-27 and M-28

[0286]

[0287] Step 1: HATU (161.9 mg, 0.43 mmol), N,N-diisopropylethylamine (127.0 mg, 0.98 mmol) and ammonium chloride (22.8 mg, 0.43 mmol) were added to a dichloromethane (5.0 mL) solution containing M-17 (00.0 mg, 0.33 mmol, 1.00 eq) prepared in Part 11 of this Example, and the reaction solution was stirred at room temperature for 16 hours. The reaction solution was concentrated and separated by thin layer preparative chromatography (dichloromethane / methanol = 10 / 1) to obtain a white solid M-26 (60.0 mg, 0.26 mmol, yield 80.2%).

[0288] M-26:MS m / z(ESI):305.2[M+H] + ;1H NMR(400MHz,DMSO-d6)δ7.32-7.35(m,2H),7.12(s,1H),7.05(s,1H),6.71(s,1H),3.96(s, 3H),3.87(s,3H),3.78(s,3H),3.23-3.32(m,1H),2.80-2.85(m,2H),1.09(d,J=6.8Hz,3H).

[0289] Step 2: At -10 ° C, trifluoroacetic anhydride (66.3 mg, 0.32 mmol) was added dropwise to a tetrahydrofuran (5.0 mL) solution containing M-26 (80.0 mg, 0.26 mmol) and pyridine (104.0 mg, 1.32 mmol). The reaction solution was slowly warmed to room temperature and stirred for 3 hours. The reaction solution was quenched with saturated sodium bicarbonate (10.0 mL), then extracted with dichloromethane (10.0 mL × 3), dried over anhydrous sodium sulfate, filtered and concentrated, and then purified using thin layer preparative chromatography (petroleum ether: ethyl acetate = 3: 1) to obtain a white solid M-27 (60.0 mg, 0.21 mmol, yield 79.7%).

[0290] Step 3: Tetrabutylammonium fluoride (661.2 mg, 2.10 mmol) and trimethylsilyl azide (241.4 mg, 2.10 mmol) were added to a solution of M-27 (30.0 mg, 0.11 mmol) in tetrahydrofuran (3.0 mL). The reaction was stirred at 100 ° C for 20 hours under sealed conditions. It was then concentrated and purified using reverse phase chromatography (C18; 0.1% formic acid aqueous solution / acetonitrile) and thin layer preparative chromatography (DCM / MeOH=20 / 1) to give a white solid M-28 (10.0 mg, 24.29 umol, yield 28.9%).

[0291] LCMS: retention time: 1.291 min; MS m / z (ESI): 330.0 [M+H] + ;

[0292] 1 H NMR(400MHz,DMSO-d6)δ16.12(br.s,1H),7.41(s,1H),7.12(s,1H),7.05(s,1H),3.94(s,3H),3.87( s,3H),3.78(s,3H),3.68-3.75(m,1H),3.50-3.55(m,1H),3.34-3.38(m,1H),1.36(d,J=6.8Hz,3H).

[0293] 18. Synthesis of compounds M-29 and M-30

[0294]

[0295] Step 1: Aluminum chloride (834 mg, 6.26 mmol,) was added to a DCM (10.0 mL) solution containing M-17 (200 mg, 0.63 mmol) prepared in Part 11 of this Example, and the resulting mixture was stirred at room temperature for 16 hours. The mixture was then carefully treated with water (50.0 mL), extracted with ethyl acetate (20.0 mL×3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by a preparative plate (EA / PE=1 / 3) to obtain M-29 (25.0 mg, 81.88 umol, 13.1% yield) and M-30 (150.0 mg, 491.28 umol, 78.5% yield).

[0296] M-29 / M-30: LCMS: retention time: 1.13min, 1.18min; MS m / z (ESI): 306.1 [M+H] + ;

[0297] M-29: 1 H NMR(400MHz,DMSO-d6)δ8.72(s,1H),δ7.31(s,1H),7.00(s,1H),6.97(s,1H),3.96(s,3H),3.88(s ,3H),3.59(s,3H),3.29-3.35(m,1H),3.06-3.11(m,1H),2.93-2.97(m,1H),1.16(d,J=7.2Hz,3H).

[0298] M-30: 1H NMR(400MHz,DMSO-d6)δ9.35(s,1H),δ7.35(s,1H),7.09(s,1H),6.83(s,1H),3.86(s,3H),3.80(s ,3H),3.59(s,3H),3.27-3.30(m,1H),3.04-3.08(m,1H),2.92-2.96(m,1H),1.16(d,J=7.2Hz,3H).

[0299] 19. Synthesis of M-31 and M-32 compounds

[0300]

[0301] Step 1: M-30 (80.0 mg, 262.02 umol) prepared in Part 18 of this Example and potassium hydroxide (29.4 mg, 524.03 umol) were dissolved in water (0.2 mL) and acetonitrile (2.0 mL), and then 1-((bromo(difluoro)methyl)-ethoxyphosphoryl)oxyethane (111.94 mg, 419.23 umol, 1.60 eq) was added. The resulting mixture was stirred at room temperature for 3 hours, water was added to the reaction solution and extracted with ethyl acetate, dried over anhydrous sodium sulfate, filtered and concentrated, and purified using thin layer preparative chromatography (ethyl acetate / petroleum ether=1 / 3) to obtain M-31 (50.0 mg, 140.71 umol, yield 53.7%).

[0302] M-31:MS m / z(ESI):356.1[M+H] + .

[0303] Step 2: M-31 (23.0 mg, 64.73 umol, 1.00 eq) was dissolved in water (0.5 mL) and methanol (2.0 mL), and then lithium hydroxide monohydrate (8.2 mg, 194.18 umol, 4.00 eq) was added. The resulting mixture was stirred at room temperature for 16 hours. The mixture was adjusted to pH <3 with dilute hydrochloric acid (1 M), and purified by high performance liquid preparative chromatography (C18; mobile phase A: water, mobile phase B: acetonitrile) to obtain a white solid M-32 (11.0 mg, 32.23 umol, yield 49.8%).

[0304] M-32:MS m / z(ESI):342.1[M+H] +.1H NMR (400MHz, DMSO-d6)7.45(d,J=2.8Hz,2H),7.34(s,1H),7.13(t,J=74.8Hz,1H),3.95(s,3H) ,3.85(s,3H),3.33-3.38(m,1H),3.01-3.08(m,1H),2.84-2.93(m,1H),1.17(d,J=7.2Hz,3H). 19 F NMR(377MHz,DMSO-d6)δ-81.29.

[0305] 20. Synthesis of M-33 compound

[0306]

[0307] Lithium hydroxide monohydrate (16.5 mg, 0.393 mmol) was added to a mixed solution of methanol (2.0 mL) and water (1.0 mL) containing M-30 (30.0 mg, 0.0983 mmol) prepared in Part 18 of this Example. The resulting mixed solution was stirred at room temperature for 16 hours. The pH of the mixed solution was adjusted to 3 with dilute hydrochloric acid, and purified by Perp-HPLC (Column: Waters Xbridge 20*150mm 10um, C18, Mobile Phase A: water, B: ACN) to obtain a white solid M-33 (20.0 mg, 68.66 umol, 69.8% yield).

[0308] M-33:MS m / z(ESI):292.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ12.15(br.s,1H),δ9.38(br.s,1H),δ7.33(s,1H),7.09(s,1H),6.83( s,1H),3.87(s,3H),3.80(s,3H),3.25-3.30(m,1H),2.83-2.98(m,2H),1.14(d,J=7.2Hz,3H).

[0309] 21. Synthesis of M-34 compound

[0310]

[0311] Lithium hydroxide monohydrate (5.5 mg, 0.13 mmol) was added to a mixed solution of methanol (1.0 mL) and water (0.5 mL) containing M-29 (10.0 mg, 32.75 umol) obtained in Part 18 of this Example. The resulting mixed solution was stirred at room temperature for 16 hours. The pH of the mixed solution was adjusted to 3 with dilute hydrochloric acid, and purified by Perp-HPLC (Column: Waters Xbridge 20*150mm 10um, C18, Mobile Phase A: water, B: ACN) to obtain a white solid M-34 (8.0 mg, 27.46 umol, 83.8% yield).

[0312] M-34:MS m / z(ESI):292.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ12.13(br.s,1H),δ8.72(s,1H),δ7.29(s,1H),7.00(s,1H),6.96(s, 1H),3.94(s,3H),3.88(s,3H),3.25-3.32(m,1H),2.86-2.98(m,2H),1.14(d,J=7.2Hz,3H).

[0313] 22. Synthesis of M-35 compound

[0314]

[0315] Step 1: Boron tribromide (dissolved in 1M dichloromethane, 13.8 mL, 13.78 mmol) was added to a dichloromethane (10.0 mL) solution containing M-17 (440.0 mg, 1.38 mmol) prepared in Part 11 of this Example at -78 ° C, and the mixture was stirred at room temperature for 3 hours. The mixture was carefully quenched with water (20.0 mL), extracted with dichloromethane (20.0 mL×2), dried over anhydrous sodium sulfate, filtered, concentrated and purified by Perp-HPLC (Column: Waters Xbridge 20*150mm 10um, C18, Mobile Phase A: water, B: ACN) to give a white solid M-35-int-1 (230.0 mg, 0.79 mmol, yield 57.6%).

[0316] M-35-int-1:MS m / z(ESI):292.1[M+H] + .

[0317] Step 2: Potassium carbonate (56.9 mg, 0.41 mmol) and 1,3-dibromopropane (104.0 mg, 0.52 mmol) were added to a DMF (2.0 mL) solution containing M-35-int-1 (30.0 mg, 0.10 mmol), and the resulting mixture was stirred at 75 ° C for 16 hours. The mixture was diluted with water (20.0 mL), extracted with ethyl acetate (10.0 mL×3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by Perp-HPLC (Column: Waters Xbridge 20*150mm 10um, C18, MobilePhase A: water, B: ACN) to give a white solid M-35 (5.75 mg, 0.0173 mmol, yield 10.5%).

[0318] M-35:MS m / z(ESI):332.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.29(s,1H),δ7.21(s,1H),6.98(s,1H),4.16-4.24(m,4H),3.97(s,3H), 3.46(s,3H),3.40-3.46(m,1H),3.00-3.12(m,2H),2.21(t,J=4.8Hz,2H),1.27(d,J=6.8Hz,3H).

[0319] 23. Synthesis of M-36 compound

[0320]

[0321] Potassium carbonate (56.9 mg, 411.95 umol) and 3-bromopropane-1-ol (42.9 mg, 308.96 umol) were added to a DMF (2.0 mL) solution containing M-35-int-1 (30.0 mg, 102.99 umol) prepared in step 22 of this example to obtain a mixture, and the mixture was stirred at 80° C. for 16 hours. The mixture was diluted with water (20.0 mL), extracted with ethyl acetate (10.0 mL×3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by Perp-HPLC (Column: Waters Xbridge 20*150mm 10um, C18, Mobile Phase A: water, B: ACN, 0.1% FA) to obtain a colorless oily liquid M-36 (1.22 mg, 2.99 umol, yield 2.9%).

[0322] M-36:MS m / z(ESI):408.3[M+H]+ ; 1 H NMR (400MHz, CDCl3) δ7.21(s,1H),7.06(s,1H),6.75(s,1H),4.26-4.29(t,J=5.8Hz,2H),4.21-4.24(t,J=5.6Hz,2H),4.01(s,3H), 3.88-3.92(m,4H),3.71(s,3H),3.39-3.45(m,1H),3.11-3.13(m,1H),2.99-3.04(m,1H),2.09-2.15(m,4H),1.27(d,J=6.8Hz,3H).

[0323] 24. Synthesis of M-37 and M-38 compounds

[0324]

[0325] Step 1: At 0°C, sodium hydride (14.0 mg, 351.60 umol, 60% in paraffin oil) was added to a tetrahydrofuran (5.0 mL) solution containing M-16-int-2 (0.1 g, 293.00 umol) prepared in step 11 of this example, and the reaction solution was stirred at 0°C for 30 minutes. Then, methyl benzoylformate (72.15 mg, 439.50 umol) was added dropwise to the reaction solution at 0°C and then warmed to room temperature and stirred for 2 hours. The reaction solution was quenched with water (20.0 mL) and extracted with ethyl acetate (10.0 mL×2). The organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain a bright yellow solid M-37-int-1 (0.1 g, 263.57 umol, yield 89.96%).

[0326] Step 2: Pd / C (10% carbon wetted with about 55% water, 10.0 mg) was added to a solution of M-37-int-1 (80 mg, 210.86 umol) in methanol (2.0 mL). The reaction solution was stirred at room temperature for 3 hours under a hydrogen balloon. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated and purified by chromatography on a silica gel column (petroleum ether: ethyl acetate = 10:1) to obtain a white solid M-37 (40 mg, 104.87 umol, 49.74% yield).

[0327] M-37:MS m / z(ESI):382.1[M+H] + ; 1H NMR(400MHz, Methanol-d4)δ7.38-7.24(m,6H),6.94(s,1H),4.23(dd,J=10.2,4.6H z,1H),3.99(s,3H),3.93(s,3H),3.91-3.80(m,4H),3.67(s,3H),3.39-3.33(m,1H).

[0328] Step 3: Add lithium hydroxide (7.53 mg, 314.61 umol) to a mixed solution of methanol (2.0 mL) and water (1.0 mL) containing M-37 (30.0 mg, 78.65 umol). Stir the reaction solution at room temperature for 16 hours. The reaction solution was adjusted to pH <3 using dilute hydrochloric acid, and then purified by high performance liquid chromatography (C18, mobile phase A: water, mobile phase B: acetonitrile) to obtain a white solid M-38 (26.30 mg, 71.59 umol, yield 91.01%).

[0329] M-38:MS m / z(ESI):368.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.44(s,1H),7.42-7.31(m,4H),7.31-7.23(m,1H),7.09(s,1H),7.06(s,1H),4 .11(dd,J=10.2,4.6Hz,1H),3.96(s,3H),3.87(s,3H),3.84-3.73(m,4H),3.24(dd,J=17.2,4.8Hz,1H).

[0330] 25. Synthesis of M-39 compound

[0331]

[0332] Step 1: At 0°C, M-16-int-2 (200.0 mg, 0.57 mmol) prepared in step 11 of this example was dissolved in tetrahydrofuran (4.0 mL) and then sodium hydride (20.2 mg, 0.88 mmol, 60% in paraffin liquid) was added. Under argon protection, the mixture was stirred at 0°C for 30 minutes. Then ethyl 3,3,3-trifluoropyruvate (149.5 mg, 0.88 mmol) was added dropwise at 0°C. Under argon protection, the mixture was stirred at room temperature for 1 h. It was then quenched with saturated ammonium chloride (10.0 mL), diluted with water (5.0 mL), and extracted with ethyl acetate (10.0 mL×3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, concentrated in vacuo, and purified by silica gel column (ethyl acetate / hexane=10:1) to give a white solid M-39-int-1 (74.0 mg, 0.19 mmol, 32.8% yield).

[0333] M-39-int-1:MS m / z(ESI):386.0[M+H] + .

[0334] Step 2: M-39-int-1 (60.0 mg, 0.15 mmol) and Pd / C (6.0 mg, 10%) were suspended in methanol (4.0 mL) and stirred for 4 hours under a hydrogen atmosphere at room temperature. The reaction was then detected by LCMS. The reactants were filtered and concentrated in vacuo. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate=20:1) to give M-39 (30.9 mg, 0.079 mmol, 51.2% yield) as a white solid.

[0335] M-39:MS m / z(ESI):388.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.52(s,1H),7.10(s,1H),7.07(s,1H),4.20-4.17(m,2H)3.99-3.95( m,4H),3.87(s,3H),3.79(s,3H),3.68-3.61(m,1H),3.50-3.45(m,1H),1.19(t,J=8.0Hz,1H); 19 F NMR(377MHz,DMSO-d6)δ-66.37(s).

[0336] 26. Synthesis of M-40 and M-41 compounds

[0337]

[0338] Step 1: Sodium hydride (46.9 mg, 1.95 mmol, 60% in paraffin oil) was added to a tetrahydrofuran (4.0 mL) solution of M-16-int-2 (0.2 g, 586.00 umol) at 0°C, and the reaction solution was stirred at 0°C for 30 minutes, and then ethyl 2-oxo-3-methylbutyrate (169.0 mg, 1.17 mmol) was added dropwise to the reaction solution. The reaction solution was quenched with water (20.0 mL) and extracted with ethyl acetate (10.0 mL×2). The organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 8 / 1) to obtain a yellow oil M-40-int-1 (0.12 g, 333.88 umol, yield 57.0%).

[0339] M-40-int-1:MS m / z(ESI):360.1[M+H] + ;

[0340] Step 2: Pd / C (10% carbon wetted with about 55% water, 20.0 mg) was added to a solution of M-40-int-1 (.1 g, 278.23 umol) in methanol (2.0 mL). The reaction solution was stirred at 40°C for 1 hour under a hydrogen balloon. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated and purified by chromatography on a silica gel column (petroleum ether / ethyl acetate = 3 / 1) to obtain a white solid M-40 (96.0 mg, 265.61 umol, yield 95.5%).

[0341] M-40:MS m / z(ESI):362.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.44(s,1H),7.11(s,1H),7.05(s,1H),4.05(m,2H),3.94(s,3H),3.87(s,3H),3.78(s,3H),3.30-3.25(m ,1H),3.05(dd,J=17.0,3.6Hz,1H),2.80-2.70(m,1H),1.96(dd,J=13.2,6.6Hz,1H),1.15(t,J=7.2Hz,3H),0.95(t,J=6.8Hz,6H).

[0342] Step 3: Add lithium hydroxide (58.1 mg, 1.38 mmol) to a mixed solution of M-40 (40.0 mg, 115.14 umol) in tetrahydrofuran (1.0 mL), methanol (0.5 mL) and water (0.5 mL). Stir the reaction solution at room temperature for 16 hours. The reaction solution was adjusted to pH <3 using dilute hydrochloric acid, extracted with ethyl acetate (5.0 mL x 3), dried over anhydrous sodium sulfate and filtered, and concentrated to obtain a white solid M-41 (21.1 mg, 63.41 umol, yield 55.1%).

[0343] M-41:MS m / z(ESI):334.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ12.06(br.s,1H),7.42(s,1H),7.12(s,1H),7.05(s,1H),3.95(s,3H),3.87(s,3H),3.78(s,3H) ,3.33(d,J=8.4Hz,1H),2.97(d,J=17.2Hz,1H),2.78-2.69(m,1H),1.99(dd,J=12.8,6.6Hz,1H),0.95(d,J=6.8Hz,6H).

[0344] 27. Synthesis of M-42 and M-43 compounds

[0345]

[0346] Step 1: Sodium hydride (42.2 mg, 1.05 mmol, 60% in paraffin oil) was added to a solution of M-16-int-2 (300.0 mg, 0.88 mmol, 1.00 eq) in tetrahydrofuran (5.0 mL) at 0°C, and the reaction solution was stirred at 0°C for 30 minutes. Then ethyl 2-butyric acid ester (172.0 mg, 1.32 mmol) was added dropwise to the reaction solution at 0°C and then warmed to room temperature and stirred for 2 hours. The reaction solution was quenched with water (20.0 mL) and extracted with ethyl acetate (10.0 mL×2). The organic phases were combined and dried over anhydrous sodium sulfate, filtered and concentrated. The crude product was purified by silica gel column chromatography (petroleum ether: ethyl acetate = 8:1) to obtain a bright yellow solid M-42-int-1 (265.0 mg, 0.764 mmol, yield 87.3%).

[0347] M-42-int-1:MS m / z(ESI):346.1[M+H] + .

[0348] Step 2: Pd / C (10% carbon wetted with about 55% water, 27.0 mg) was added to a solution of M-42-int-1 (265.0 mg, 0.77 mmol) in methanol (5.0 mL). The reaction solution was stirred at room temperature for 1 hour under a hydrogen balloon. The reaction solution was filtered through diatomaceous earth, and the filtrate was concentrated and purified by chromatography on a silica gel column (petroleum ether: ethyl acetate = 10:1) to obtain a white solid M-42 (200.0 mg, 0.58 mmol, yield 75.0%).

[0349] M-42:MS m / z(ESI):348.2[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.40(s,1H),7.11(s,1H),7.05(s,1H),4.00-4.10(m,2H)3.95(s,3H),3.87(s,3H),3.78(s,3H), 3.26-3.31(m,1H),3.05-3.10(m,1H),2.78-2.82(m,1H),1.57-1.64(m,2H),1.15(t,J=7.2Hz,3H),0.91(t,J=7.2Hz,3H).

[0350] Step 3: Lithium hydroxide monohydrate (19.3 mg, 0.46 mmol) was added to a mixed solution of M-42 (40.0 mg, 115.14 umol,) in methanol (2.0 mL) and water (1.0 mL). The reaction solution was stirred at room temperature for 16 hours. The reaction solution was adjusted to pH <3 using dilute hydrochloric acid and then purified by high performance liquid chromatography (C18, A: water, B: acetonitrile) to obtain a white solid M-43 (15.0 mg, 46.97 umol, yield 40.8%).

[0351] M-43:MS m / z(ESI):320.0[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ12.06(br.s,1H),7.39(s,1H),7.11(s,1H),7.05(s,1H),3.96(s,3H),3.87(s,3H),3. 78(s,3H),3.25-3.32(m,1H),2.97-3.01(m,1H),2.74-2.78(m,1H),1.55-1.64(m,2H),0.92(t,J=7.2Hz,3H).

[0352] 28. Synthesis of M-44 compound

[0353]

[0354] Step 1: Dissolve 5,6-dimethoxyindole-2-carboxylic acid ethyl ester (2.0 g, 8.02 mmol) in a solution of tetrahydrofuran / N,N-dimethylformamide = 4 / 1 (50.0 mL) and add sodium hydride (481.42 mg, 12.04 mmol, 60% dispersed in paraffin liquid), stir for 0.5 hours at 0 ° C and argon protection. Add 2-(trimethylsilyl)ethoxymethyl chloride (16.0 g, 9.63 mmol) to the reaction mixture and stir at room temperature for 2 hours. The reaction mixture was then quenched with saturated ammonium chloride (50.0 mL) and extracted with ethyl acetate (50.0 mL×3). Dry with anhydrous sodium sulfate, filter and concentrate in vacuo, and purify by silica gel column chromatography (petroleum ether: ethyl acetate = 5: 1) to give a white solid M-44-int-1 (2.3 g, 6.06 mmol, yield 75.7%).

[0355] Step 2: At -78 ° C, under nitrogen protection, bis(trimethylsilyl)lithium aza (7.91 mL, 7.91 mmol, 1M in tetrahydrofuran) was added to a solution of M-44-int-1 (2.0 g, 5.27 mmol) in tetrahydrofuran (40.0 mL) and stirred for 0.5 h. Dimethyl methylphosphonate (1.0 g, 8.06 mmol) was then added to the reaction mixture and stirred at room temperature for 16 h. The reaction mixture was subsequently quenched with saturated ammonium chloride (10.0 mL), diluted with water (30.0 mL), extracted with EtOAc (30.0 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (PE: EA = 1: 1) to give a yellow solid M-44-int-2 (2.0 g, 4.37 mmol, 83.0% yield).

[0356] M-44-int-2: 1 H NMR (400MHz, CDCl3) δ7.31 (d, J=1.6Hz, 1H), 7.03 (d, J=2.4Hz, 1H), 6.92 (d, J=

[0357] 2.0Hz,1H),5.99(d,J=2.4Hz,2H),3.96(d,J=2.4Hz,3H),3.91(d,J=2.8Hz,3H),3.83-3.78(m,3H) ,3.77-3.73(m,3H),3.58-3.56(m,2H),3.54-3.48(m,2H),0.88-0.83(m,2H),-0.07--0.21(m,9H).

[0358] Step 3: M-44-int-2 (1.2 g, 2.62 mmol, 1.00 eq) was dissolved in tetrahydrofuran (20.0 mL) and sodium hydride (105.0 mg, 2.63 mmol, 60% paraffin liquid) was added and stirred for 0.5 hours at 0 ° C under argon protection. Then methyl 2-oxopropanoate (401.3 mg, 3.93 mmol) was added to the reaction mixture and stirred at room temperature for 2 h. The reaction was then quenched with saturated ammonium chloride (10.0 mL), diluted with water (20.0 mL) and extracted with ethyl acetate (20.0 mL × 3), dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain M-44-int-3 (500.0 mg, 1.15 mmol, 43.9% yield) as a yellow oil by purification by silica gel column chromatography (petroleum ether / ethyl acetate = 5: 1).

[0359] M-44-int-3:MS m / z(ESI):456.3[M+Na] + .

[0360] Step 4: M-44-int-3 (1.0 mg, 2.31 mmol) was dissolved in MeOH (8.0 mL) and Pd / C (100.0 mg, 10% wt, 50% water) was added and stirred at 25 ° C for 2 hours under a hydrogen balloon. After the reaction was completed, the mixture was filtered and concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 5: 1) to give compound M-44-int-4 (600.0 mg, 1.38 mmol, yield 60.0%) as a white solid.

[0361] M-44-int-4:MS m / z(ESI):458.3[M+H] + .

[0362] Step 5: M-44-int-4 (20.0 mg, 0.04 mmol) was dissolved in tetrahydrofuran (3.0 mL) at room temperature, and tetrabutylammonium fluoride (24.01 mg, 0.09 mmol) was added, and the reaction mixture was stirred at 60 ° C for 16 hours. After the reaction was completed, the reaction mixture was diluted with water (10.0 mL), and extracted with ethyl acetate (10.0 mL × 3), and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by thin layer preparative chromatography (petroleum ether / ethyl acetate = 3 / 1) to obtain M-44 (1.63 mg, 0.005 mmol, yield 11.6%) as a white solid.

[0363] M-44:MS m / z(ESI):318.0[M-18+H] + ; 1H NMR(400MHz,MeOD)δ7.55(s,1H),7.28(s,1H),7.18(s,1H),6.10(s,2H),4.04(s,3H),3.98( s,3H),3.78(s,3H),3.52(dd,J=16.4,8.0Hz,1H),3.26-3.09(m,2H),1.36(d,J=7.2Hz,3H).

[0364] 29. Synthesis of M-45 and M-46 compounds

[0365]

[0366] Step 1: Dissolve tert-butyl acetate (1.32 g, 11.39 mmol) in THF (20.0 mL) and add LiHDMS (11.4 mL, 11.39 mol, 1 M in THF) at -78 °C under nitrogen protection. Stir the mixture at this temperature for 30 minutes. Then add ethyl 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylate (1.0 g, 3.80 mmol) dissolved in tetrahydrofuran (10.0 mL). Warm the mixture to room temperature and stir for 2 hours. The mixture was treated with aqueous ammonium chloride solution (50.0 mL), extracted with DCM (50.0 mL×2), and purified by silica gel column (PE / EA=3 / 1) to give tert-butyl 3-(5,6-dimethoxy-1-methylindol-2-yl)-3-oxopropanoate M-45-int-1 (1.1 g, 3.30 mmol, yield 86.9%) as a white solid.

[0367] M-45-int-1:MS m / z(ESI):278.1[M-56+H] + .

[0368] Step 2: M-45-int-1 (1.0 g, 3.00 mmol) was dissolved in DCM (10.0 mL) and 2,2,2-trifluoroacetic acid (2.98 g, 26.14 mmol, 2 mL) was added. The mixture was stirred at room temperature for 16 hours. The solvent was removed in vacuo to give crude M-45-int-2 (830.0 g, 2.99 mmol, 99.8% yield) as a brown oil. The crude product was used directly in the next step without purification.

[0369] M-45-int-2:MS m / z(ESI):278.1[M+H] + .

[0370] Step 3: M-45-int-2 (830 mg, 2.99 mmol) was dissolved in toluene (10.0 mL) and stirred at 80° C. for 1 hour. The mixture was concentrated in vacuo and purified by a silica gel column (petroleum ether / ethyl acetate=5 / 1) to give 1-(5,6-dimethoxy-1-methylindol-2-yl)ethanone M-45-int-3 (680.0 mg, 2.92 mmol, 97.4% yield) as a white solid.

[0371] M-45-int-3:MS m / z(ESI):234.1[M+H] + .

[0372] Step 4: Sodium metal (19.7 mg, 857.40 umol) was added to methanol (2.0 mL) and stirred at room temperature for 10 minutes. Then M-45-int-3 (50.0 mg, 214.35 umol) was added. The mixture was stirred for another 30 minutes. Then diethyl oxalate (46.99 mg, 321.53 umol) was added and stirred at 80 ° C for 2 hours. The mixture was concentrated and purified by thin layer preparative chromatography (dichloromethane / methanol=20 / 1) to obtain (Z)-4-(5,6-dimethoxy-1-methylindol-2-yl)-2-hydroxy-4-oxobutyl-2-enoic acid methyl ester M-45-int-4 (50.0 mg, 156.59 umol, yield 73.1%) as a yellow solid.

[0373] M-45-int-4:MS m / z(ESI):320.2[M+H] + .

[0374] Step 5: M-45-int-4 (120.0 mg, 375.81 umol) was dissolved in tetrahydrofuran (5.0 mL) and methanol (2.0 mL) and diazomethyl (trimethyl) silane (85.9 mg, 751.62 umol, 2M in hexane) was added. The mixture was stirred at 25 ° C for 16 hours. The mixture was concentrated and purified by thin layer preparative chromatography (petroleum ether / ethyl acetate = 2 / 1) to give a yellow solid (Z) -4- (5,6-dimethoxy-1-methylindol-2-yl) -2-methoxy-4-oxobutyl-2-enoic acid methyl ester M-45-int-5 (88.0 mg, 264.00 umol, yield 70.3%).

[0375] M-45-int-5:MS m / z(ESI):334.0[M+H] + .

[0376] Step 6: M-45-int-5 (80.0 mg, 30.00 umol) was dissolved in methanol (3.0 mL) and Pd / C (10%, 8.0 mg) was added. The mixture was stirred at 25 ° C under H2 for 16 hours. The mixture was filtered and concentrated, and purified by thin layer preparative chromatography (petroleum ether / ethyl acetate=2 / 1) to give a white solid M-45 (35.0 mg, 104.37 umol, yield 43.5%).

[0377] M-45:MS m / z(ESI):336.3[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.39(s,1H),7.10(s,1H),7.06(s,1H),4.32(dd,J=7.6,4.8Hz,1H),3.97 (s,3H),3.87(s,3H),3.78(s,3H),3.33-3.43(m,1H),3.31(s,3H),3.23(dd,J=16.4,4.8Hz,1H).

[0378] Step 7: M-45 (20.0 mg, 59.6 umol) was dissolved in water (1.0 mL) and methanol (2.0 mL) and then lithium hydroxide monohydrate (10.01 mg, 238.56 umol) was added. The mixture was stirred at 25 ° C for 16 hours. Purification by high performance liquid chromatography (C18, A: 0.1% formic acid aqueous solution, B: acetonitrile) gave a white solid M-46 (2.38 mg, 7.41 umol, yield 12.4%).

[0379] M-46:MS m / z(ESI):322.2[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.38(s,1H),7.11(s,1H),7.06(s,1H),4.16-4.45(m,1H ),3.98(s,3H),3.87(s,3H),3.78(s,3H),3.28-3.29(m,4H),3.08-3.20(m,1H).

[0380] 30. Synthesis of compounds M-47 and M-48

[0381]

[0382] Step 1: At 0°C, sodium hydride (7.5 g, 188.0 mmol) was added to a solution of N,N-dimethylformamide (200.0 mL) containing 5-methoxy-indole-2-carboxylic acid (9.0 g, 47.0 mol), and the resulting mixture was stirred at 0°C for 10 minutes. Then iodomethane was added. Then stirred at room temperature for 16 hours, quenched with saturated ammonium chloride (50.0 mL), diluted with water (50.0 mL), extracted with ethyl acetate (100.0 mL x 2), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (10–100%, ethyl acetate / petroleum ether) to obtain crude M-47-int-1 (12.8 g, 58.4 mmol) as a white solid.

[0383] M-47-int-1:MS m / z(ESI):220.1[M+H] + .

[0384] Step 2: Under argon protection, LiHMDS (116.0 mL, 116.0 mmol, 1M in THF) was added to a tetrahydrofuran (200.0 mL) solution containing M-47-int-1 (12.8 g, 58.0 mmol) and dimethyl methylphosphonate (10.8 g, 87.0 mmol) at -78 ° C, and then slowly warmed to room temperature and stirred for 3 hours. The reaction was quenched with saturated ammonium chloride (50.0 mL), diluted with water (50.0 mL), extracted with ethyl acetate (100.0 mL x 2), and the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (10-100%, ethyl acetate / petroleum ether) to obtain crude M-47-int-2 (15.0 g, 48.0 mmol, yield 82.8%) as a white solid.

[0385] M-47-int-2:MS m / z(ESI):312.1[M+H].

[0386] Step 3: Sodium hydride (60% in paraffin oil, 2.5 g, 62.55 mmol, 1.50 eq) was added to a tetrahydrofuran (200.0 mL) solution containing M-47-int-2 (13.0 g, 41.70 mmol, 1.00 eq) at 0 ° C and stirred for 30 minutes at 0 ° C. Methyl pyruvate (6.39 g, 62.55 mmol, 1.50 eq) was then added dropwise to the reaction solution, warmed to room temperature and stirred for 1 hour. Then saturated ammonium chloride (50.0 mL) was quenched, diluted with water (50.0 mL), extracted with ethyl acetate (100.0 mL x2), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (10-100% ethyl acetate / petroleum ether) to obtain a yellow solid M-47-int-3 (16.0 g, 55.68 mmol, crude product).

[0387] M-47-int-3:MS m / z(ESI):288.1[M+H] + .

[0388] Step 4: M-47-int-3 (15.0 g, 52.0 mmol, 1.00 eq) was dissolved in methanol (20.0 mL), and then Pd / C (1.0 g, 10%) was added, and stirred at 40°C for three hours under a hydrogen balloon. The reaction was monitored by LCMS and the raw material was found to be small. The reaction solution was filtered, and the filtrate was concentrated and purified by silica gel chromatography (petroleum ether: ethyl acetate = 20: 1) to obtain white solid M-47 (5.0 g, 0.017 mol, yield 33.1%) and white solid M-48 (40.71 mg).

[0389] M-47:MS m / z(ESI):290.1[M+H] + ;

[0390] 1 H NMR (400MHz, DMSO-d6) δ7.49(d,J=12.0Hz,1H),7.43(s,1H),7.14(d,J=2.4Hz,1H),7.03(dd,J=2.4Hz,1H),3.95( s,3H),3.79(s,3H),3.60(s,3H),3.31-3.36(m,1H),3.07-3.17(m,1H),2.93-3.00(m,1H),1.18(d,J=12.0Hz,3H).

[0391] M-48:MS m / z(ESI):304.1[M+H] + ; 1H NMR(400MHz,DMSO-d6)δ7.43(d,J=9.2Hz,1H),7.17(d,

[0392] J=2.4Hz,1H),7.01(dd,J=9.2,2.4Hz,1H),3.82(d,J=7.6Hz,6H),3.61(s,3H),3.36-3.25( m,1H),3.10(dd,J=17.6,4.8Hz,1H),3.05-2.96(m,1H),2.58(s,3H),1.21(d,J=7.2Hz,3H).

[0393] 31. Synthesis of M-49 compound

[0394]

[0395] M-47 (100.0 mg, 0.35 mmol) and lithium hydroxide (30.0 mg, 0.69 mmol) prepared in part 31 of this example were dissolved in a mixed solution of tetrahydrofuran and water (v / v=2 / 1, 2.0 mL) and stirred at 40°C for 16 hours. The resulting reaction solution was adjusted to pH <3 with dilute hydrochloric acid (1N), the reaction solution was diluted with water (10.0 mL) and extracted with ethyl acetate (10.0 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated. The reaction residue was purified by silica gel column chromatography (dichloromethane / methanol=10 / 1) to obtain a white solid M-49 (21.9 mg, 0.079 mmol, yield 22.0%).

[0396] M-49:MS m / z(ESI):276.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.49 (d, J=9.2Hz, 1H), 7.41 (s,

[0397] 1H),7.15(d,J=2.4Hz,1H),7.02(dd,J=9.2,2.4Hz,1H),3.96(s,3H),3.78(s,3H),3.25–3.4 3(m,7H),3.03(dd,J=16.8,5.4Hz,1H),2.88(dd,J=13.6,7.4Hz,1H),1.17(d,J=7.2Hz,3H).

[0398] 32. Synthesis of M-50 compound

[0399]

[0400] Boron tribromide (6.92 mL, 6.92 mmol, 1 M dichloromethane solution) was added dropwise to a dichloromethane (2.0 mL) solution containing M-47 (200.0 mg, 0.69 mmol) prepared in Part 30 of this Example at -78 °C, then slowly heated to 0 °C and stirred for 2 hours under argon protection. The reaction solution was quenched with methanol and diluted with water (10.0 mL), and extracted with ethyl acetate (10.0 mL x 3), the organic phase was dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1: 1) to obtain a white solid M-50 (4.98 mg, 0.018 mmol, 2.6% yield).

[0401] M-50:MS m / z(ESI):298.1[M+Na] + ; 1 H NMR (400MHz, CDCl3) δ7.24 (s, 1H), 7.19 (s, 1H), δ7.05 (d, J = 2.4Hz, 1H), 6.98 (dd, J = 8.9, 2.4H z,1H),4.01(s,3H),3.71(s,3H),3.48-3.42(m,1H),3.14-3.01(m,2H),1.28(d,J=8.0Hz,1H).

[0402] 33. Synthesis of M-51 compound

[0403]

[0404] 250 mg of ethyl 5,6-dimethoxyindole-2-carboxylate (1.0 mmol) was dissolved in 5.0 mL of THF, and 224 mg of potassium tert-butoxide (2.0 mmol) and 150 mg of ethyl acrylate (1.5 mmol) were added in sequence, and the temperature was raised to 70°C for 12 hours. The reaction solution was cooled to room temperature, most of the solvent was evaporated, and the residue was poured into 5.0 mL of ice water, and the pH was adjusted to 1-2 with 1N hydrochloric acid aqueous solution, extracted with ethyl acetate, and the organic layers were combined, dried, concentrated, and column chromatography was performed to obtain 90 mg of M-51.

[0405] M-51:purity:96%; 1 H NMR(400MHz,DMSO-d6)δ7.18(s,1H),7.17(s,1H),6.94(s,1H),4.68–4.55(m,2H),.39(dd, J=7.9,4.2Hz,1H),4.18(d,J=7.0Hz,2H),3.88(s,3H),3.80(s,3H),1.23(t,J=7.1Hz,3H).

[0406] 34. Synthesis of M-52 compound

[0407]

[0408] Step 1: Dissolve 250 mg of ethyl 5,6-dimethoxyindole-2-carboxylate (1 mmol) in 5 mL of DMF, slowly add 44 mg of NaH (1.2 mmol) under ice bath, and warm to room temperature. React at room temperature for 1 h, slowly drop 390 mg of ethyl 4-bromobutyrate (2 mmol) into the reaction solution, and continue to react at room temperature for 12 h. After the reaction is completed, add water to quench, extract with ethyl acetate, combine the organic layers, dry, concentrate, and column chromatograph to obtain 261 mg of M-52-int-1.

[0409] Step 2: Dissolve 182 mg of the intermediate M-52-int-1 (0.5 mmol) in 10.0 mL of THF, add 112 mg of potassium tert-butoxide in batches, heat to 70°C, and react for 12 h. Concentrate the reaction solution, pour the residue into water, adjust the pH to 1-2 with 1N hydrochloric acid aqueous solution, extract with ethyl acetate, combine the organic layers, dry, concentrate, and column chromatography to obtain 70 mg of a pair of enol tautomers M-52 (keto / alcohol ratio is about 4:1).

[0410] M-52-ol: 1 H NMR(400MHz,DMSO-d6)δ7.14(s,1H),7.12(s,1H),7.09(s,1H),4.46–4.38(m,1H),4. 22–4.10(m,4H),3.91–3.87(m,1H),3.87(s,3H),3.78(s,3H),1.23(t,J=7.1Hz,3H).

[0411] 35. Synthesis of M-53 compound

[0412]

[0413] Step 1: Dissolve 250 mg of ethyl 5,6-dimethoxyindole-2-carboxylate (1 mmol, 1.0 eq) in 10 mL of DMF, add 44 mg of NaH (1.2 mmol, 1.2 eq) in batches under an ice bath, and slowly warm to room temperature. After reacting at room temperature for 1 hour, slowly drop 418 mg of ethyl 5-bromo-n-valeric acid (2 mmol, 2.0 eq) into the reaction solution, and react at room temperature for 12 hours. After the reaction is complete, add water to quench, extract with ethyl acetate, combine the organic layers, dry, concentrate, and column chromatograph to obtain 245 mg of M-53-int-1.

[0414] Step 2: Dissolve 189 mg of intermediate M-53-int-1 (0.5 mmol, 1.0 eq) in 10.0 mL of THF, add 112 mg of potassium tert-butoxide (1.0 mmol, 2.0 eq) in batches, heat to 70°C, and react for 12 h. Concentrate the reaction solution, pour the residue into water, adjust the pH to 1-2 with 1N hydrochloric acid aqueous solution, extract with ethyl acetate, combine the organic layers, dry, concentrate, and column chromatography to obtain 86 mg of a pair of enol tautomers M-53 (keto / alcohol ratio is about 2:1).

[0415] M-53-ol: 1 H NMR(400MHz,DMSO-d6)δ7.27(s,1H),7.16(s,1H),7.15(s,1H),4.33–4.26(m,2H),4. 22–4.18(m,3H),3.92(s,3H),3.82(s,3H),2.26–1.95(m,4H),1.26(t,J=7.1Hz,3H).

[0416] 36. Synthesis of compounds M-54 and M-55

[0417]

[0418] Step 1: Dissolve 235 mg of 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylic acid (1 mmol, 1.0 eq), 456 mg of HATU (1.2 mmol, 1.2 eq) and 258 mg of DIPEA (2 mmol, 2.0 eq) in 10 mL of THF and react at room temperature for 1 h. Add 234 mg of methyl 3-(methylamino)propionate (2 mmol, 2.0 eq) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is complete, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 234 mg of M-54.

[0419] M-54:purity:99%;HRMS(ESI)m / z:[M+Na] + Calcd for C 17 H 22 N2NaO5 357.1426; Found 357.1423; 1HNMR(600MHz,Chloroform-d)δ7.03(s,1H),6.78(s,1H),6.54(s,1H),3.97(s,3H),3.91(s ,3H),3.87(t,J=7.0Hz,2H),3.81(s,3H),3.69(s,3H),3.20(s,3H),2.70(t,J=7.0Hz,2H).

[0420] Step 2: 100 mg M-54 (0.3 mmol) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 36 mg LiOH (1.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography to obtain 43 mg M-55.

[0421] M-55:purity:99%;HRMS(ESI)m / z:[M+H] + Calcd for C 16 H 21 N2O5 321.1450; Found321.1448; 1 HNMR(600MHz,DMSO-d6)δ12.39(br s,1H),δ7.07(s,1H),7.04(s,1H),6.53(s,1H),3.83(s,3H),3.73(s,3H),3.71(br s,2H),3.70(s,3H),3.04(s,3H),2.57(t,J=7.0Hz,2H).

[0422] 37. Synthesis of M-56 and M-57 compounds

[0423]

[0424] Step 1: Dissolve 150 mg 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylic acid (0.64 mmol), 239 mg HATU (0.77 mmol) and 330 mg DIPEA (2.56 mmol) in 10 mL THF and react at room temperature for 1 h. Add 214 mg 3-amino-2,2-dimethylpropionic acid methyl ester hydrochloride (2 mmol) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 188 mg M-56-int-1.

[0425] Step 2: Dissolve 235 mg of M-56-int-1 (0.5 mmol) in 10 mL of DMF, add 40 mg of NaH (1.0 mmol, 60% dispersed in oil) in batches under ice bath, slowly warm to room temperature, and react for 1 hour. Slowly add 234 mg of MeI (2 mmol) to the reaction solution, and continue to react for 12 hours. After the reaction is completed, add saturated aqueous ammonium chloride to the reaction solution under ice bath to quench, dilute with water, extract with ethyl acetate, combine the organic layers, dry, concentrate, and separate by column chromatography to obtain 109 mg of M-56.

[0426] M-56:purity:99%;HRMS(ESI)m / z:[M+Na] + Calcd for C 19 H 26 N2NaO5 385.1739; Found 385.1745; 1 HNMR(400MHz,DMSO-d6)δ7.07(s,1H),7.04(s,1H),6.57(s,1H),3.83(s,3H),3.75(s,3H),3.73(br s,2H),3.70(s,3H),3.60(s,3H),3.04(s,3H),1.15(s,6H).

[0427] Step 3: 109 mg M-56 (0.3 mmol) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 36 mg LiOH (1.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography (DCM / MeOH / HCOOH) to obtain 35 mg M-57.

[0428] M-57:LCMS:Rt:4.43min; purity:97%; HRMS(ESI)m / z:[M+H] + Calcd for C 16 H 21 N2O5 321.1450; Found 321.1448; 1 H NMR(600MHz,DMSO-d6)δ12.46(s,1H),7.07(s,1H),7.04(s,1H),6.56(s,1H),3.83(s,3H),3.75(s,3H),3.73(br s,2H),3.71(s,3H),3.07(s,4H),1.13(s,6H).

[0429] 38. Synthesis of compounds M-58 and M-59

[0430]

[0431] Step 1: Dissolve 470 mg of 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylic acid (2 mmol), 912 mg of HATU (2.4 mmol) and 330 mg of DIPEA (4 mmol) in 20 mL of THF and react at room temperature for 1 h. Add 612 mg of 3-amino-2-methylpropionic acid methyl ester hydrochloride (4 mmol) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 434 mg of M-58.

[0432] M-58:purity:97%;HRMS(ESI)m / z:[M+Na] + Calcd for C 17 H 22 N2NaO5 357.1426; Found 357.1422; 1 HNMR(600MHz,DMSO-d6)δ8.39(t,J=6.0Hz,1H),7.08(s,1H),7.03(s,1H),6.94(s,1H),3.93(s,3H),3.84(s,3 H),3.76(s,3H),3.61(s,3H),3.46–3.41(m,1H),3.31–3.26(m,1H),2.80–2.72(m,1H),1.10(d,J=7.0Hz,3H).

[0433] Step 2: 167 mg M-58 (0.5 mmol) was dissolved in a mixed solvent of 6.0 mL THF, 6.0 mL MeOH and 2.0 mL H2O, 60 mg LiOH (2.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, and the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography (DCM / MeOH / HCOOH) to obtain 128 mg M-59.

[0434] M-59:purity:99%;HRMS(ESI)m / z:[M+Na] + Calcd for C 16 H 20 N2NaO5 343.1270; Found 343.1266; 1 HNMR(600MHz,DMSO-d6)δ8.41(t,J=5.8Hz,1H),7.07(s,1H),7.03(s,1H),6.95(s,1H),3.94(s,3H),3.84(s,3H),3. 76(s,3H),3.41(dt,J=12.5,5.9Hz,1H),3.26(dt,J=13.0,6.3Hz,1H),2.62(q,J=7.0Hz,1H),1.07(d,J=7.0Hz,3H).

[0435] 39. Synthesis of M-60 and M-61 compounds

[0436]

[0437] Step 1: Dissolve 235 mg 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylic acid (1 mmol), 456 mg HATU (1.2 mmol) and 258 mg DIPEA (2.0 mmol) in 10 mL THF and react at room temperature for 1 h. Add 262 mg 3-amino-2,2-dimethylpropionic acid methyl ester (2 mmol) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 209 mg M-60.

[0438] M-60:purity:98%;HRMS(ESI)m / z:[M+Na] + Calcd for C 18 H 24N2NaO5 371.1583; Found 371.1582; 1 HNMR(600MHz,DMSO-d6)δ8.23(t,J=6.5Hz,1H),7.09(s,1H),7.03(s,1H),6.95(s,1H), 3.91(s,3H),3.84(s,3H),3.76(s,3H),3.61(s,3H),3.40(d,J=6.4Hz,2H),1.15(s,6H).

[0439] Step 2: 104 mg M-60 (0.3 mmol) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 36 mg LiOH (1.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography (DCM / MeOH / HCOOH) to obtain 32 mg M-61.

[0440] M-61:purity:97%;HRMS(ESI)m / z:[M+Na] + Calcd for C 17 H 22 N2NaO5 357.1426; Found 357.1425; 1 HNMR(600MHz,DMSO-d6)δ8.29(t,J=6.3Hz,1H),7.07(s,1H),7.03(s,1H),6.93(s ,1H),3.92(s,3H),3.83(s,3H),3.75(s,3H),3.35(d,J=5.9Hz,2H),1.10(s,6H).

[0441] 40. Synthesis of M-62 and M-63 compounds

[0442]

[0443] Step 1: Dissolve 150 mg 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylic acid (0.64 mmol), 239 mg HATU (0.77 mmol) and 330 mg DIPEA (2.56 mmol) in 10 mL THF and react at room temperature for 1 h. Add 197 mg 3-amino-2-methylpropionic acid methyl ester hydrochloride (1.28 mmol) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 150 mg M-62-int-1.

[0444] Step 2: Dissolve 167 mg M-62-int-1 (0.5 mmol) in 10 mL DMF, add 40 mg NaH (1.0 mmol, 60% dispersed in oil) in batches under ice bath, slowly warm to room temperature, and react for 1 hour. Slowly add 234 mg MeI (2 mmol) to the reaction solution, and continue to react for 12 hours. After the reaction is completed, add saturated ammonium chloride aqueous solution to the reaction solution under ice bath to quench, dilute with water, extract with ethyl acetate, combine the organic layers, dry, concentrate, and separate by column chromatography to obtain 115 mg M-62.

[0445] M-62:purity:99%;HRMS(ESI)m / z:[M+Na] + Calcd for C 18 H 24 N2NaO5 371.1583; Found 371.1594; 1 HNMR(600MHz,DMSO-d6)δ7.08(s,1H),7.05(s,1H),6.54(s,1H),3.84(s,3H),3 .76(s,3H),3.70(s,4H),3.64–3.60(m,2H),3.60(s,3H),3.06(s,3H),2.93(br s,1H),1.06(br s,3H).

[0446] Step 3: 105 mg M-62 (0.3 mmol) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 36 mg LiOH (1.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography (DCM / MeOH / HCOOH) to obtain 40 mg M-63.

[0447] M-63:HRMS(ESI)m / z:[MH] - Calcd for 17 H 21 N2O5 333.1450; Found 333.1458; 1 HNMR(600MHz,DMSO-d6)δ7.07(s,1H),7.04(s,1H),6.52(s,1H),3.83(s,3H),3.75(s,3H),3.69(s,3H),3.65–3.60(m,2H),3.05(s,3H),2.80(br s,1H),1.02(s,3H).

[0448] 41. Synthesis of M-64 and M-65 compounds

[0449]

[0450] Step 1: Dissolve 150 mg 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylic acid (0.64 mmol), 239 mg HATU (0.77 mmol) and 330 mg DIPEA (2.56 mmol) in 10 mL THF and react at room temperature for 1 h. Add 179 mg sarcosine methyl ester hydrochloride (1.28 mmol) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate 182 mg M-64 by column chromatography.

[0451] M-64:purity:99%;HRMS(ESI)m / z:[M+Na] + Calcd for C 16 H 20 N2NaO5 343.1270; Found 343.1274; 1 HNMR(600MHz,DMSO-d6)δ7.08(s,1H),7.04(s,1H),6.64(s,0.65H),6.42(s,0.35H),4.39(s,0.7H), 4.27(s,1.3H),3.84(s,3H),3.76(s,3H),3.72(s,3H),3.70(s,3H),3.17(s,1.95H),3.02(s,1.05H).

[0452] Step 2: 96 mg M-64 (0.3 mmol) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 36 mg LiOH (1.5 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography (DCM / MeOH / HCOOH) to obtain 51 mg M-65.

[0453] M-65:purity:95%; HRMS(ESI)m / z:[M+H] + Calcd for C 15 H 19 N2O5 307.1294; Found307.1298; 1 HNMR(600MHz,DMSO-d6)δ7.08(s,1H),7.05(s,1H),6.62(s,0.5H),6.45(s,0.5H),4.24(s ,1H),4.17(s,1H),3.84(s,3H),3.76(s,3H),3.72(s,3H),3.15(s,1.5H),3.01(s,1.5H).

[0454] 42. Synthesis of M-66 and M-67 compounds

[0455]

[0456] Step 1: Dissolve 235 mg of 5,6-dimethoxy-1-methyl-1-H-indole-2-carboxylic acid (1 mmol, 1.0 eq), 456 mg of HATU (1.2 mmol, 1.2 eq) and 516 mg of DIPEA (4 mmol, 4.0 eq) in 10 mL of THF and react at room temperature for 1 h. Add 278 mg of 3-aminopropionic acid methyl ester hydrochloride (2 mmol, 2.0 eq) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 208 mg of M-66.

[0457] M-66:purity:91%; HRMS(ESI)m / z:[M+Na] + Calcd for C 16 H 20 N2NaO5 343.1270; Found 343.1263; 1HNMR(600MHz,DMSO-d6)δ8.36(t,J=5.5Hz,1H),7.07(s,1H),7.03(s,1H),6.93(s,1H),3.94(s, 3H),3.83(s,3H),3.76(s,3H),3.61(s,3H),3.47(td,J=7.0,5.5Hz,2H),2.59(t,J=7.0Hz,2H).

[0458] Step 2: 96 mg M-66 (0.3 mmol, 1.0 eq) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 36 mg LiOH (1.5 mmol, 5.0 eq) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, and the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography (DCM / MeOH / HCOOH) to obtain 43 mg M-67.

[0459] M-67:purity:95%; HRMS(ESI)m / z:[M+H] + Calcd for C 15 H 18 N2NaO5 329.1113; Found 329.1108; 1 HNMR(600MHz,DMSO-d6)δ12.19(s,1H),8.33(t,J=5.6Hz,1H),7.07(s,1H),7.03(s,1H),6.94(s ,1H),3.94(s,2H),3.83(s,2H),3.76(s,2H),3.43(td,J=7.1,5.5Hz,2H),2.51(t,J=7.2Hz,2H)

[0460] 43. Synthesis of compounds M-68, M-69 and M-70

[0461]

[0462] 1-Chloromethyl-4-fluoro-1,4-diazotized bicyclo[2.2.2]oxane (tetrafluoroborate) (1.66 g, 4.70 mmol) was added to a solution of 10 mL acetonitrile and 10 mL dichloromethane containing M-16 (1.00 g, 3.13 mmol) prepared in Part 11 of this Example, and stirred overnight at 55 degrees Celsius. The reaction solution was extracted with ethyl acetate and water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and pre-purified with a silica gel column (20:80 ethyl acetate-petroleum ether), and further purified by preparative chromatography to obtain M-68 (11 mg, 0.033 mmol, 1% yield), M-69 (8 mg, 0.024 mmol, 0.8% yield) and M-69 (10 mg, 0.03 mmol, 0.9% yield), respectively.

[0463] M-68:HRMS(ESI)m / z:[M+H] + Calcd for C 17 H 20 FNO5 337.1326,found:338.1401; 1 HNMR(600MHz, CDCl3)δ6.98(s,1H),6.67(s,1H),3.98(s,3H),3.94(s,6H),3.72(s,3H),3.42(dd, J=17.4,8.3Hz,1H),3.15–3.07(m,1H),3.07–3.00(m,1H),1.29(d,J=7.0Hz,1H); 13CNMR(151MHz, CDCl3)δ190.71,176.40,154.35,154.32,150.36,148.70,137.18,137.10,133.70,132.31,132.2 3,110.37,110.23,107.92,87.82,87.80,62.04,62.02,56.43,51.94,42.75,35.18,32.55,17.24.

[0464] M-69:HRMS(ESI)m / z:[M+H] + Calcd for C 17 H 20 FNO5 337.1326,found:338.1404; 1HNMR(600MHz, CDCl3)δ7.33(s,1H),6.52(s,1H),4.01(s,3H),3.97(s,3H),3.94(s,3H),3.71(s,3H),3 .44(dd,J=16.7,8.0Hz,1H),3.15–3.07(m,1H),3.03(dd,J=16.8,5.4Hz,1H),1.28(d,J=7.0Hz,3H); 13C NMR (151MHz, CDCl3) δ188.76,188.73,176.58,151.76,150.71,148.98,146.26,132.21,132.18,118 .08,117.96,106.98,106.87,98.07,91.48,56.10,56.06,51.81,44.58,44.53,34.75,32.10,17.23.

[0465] M-70:HRMS(ESI)m / z:[M+H] + Calcd for C 17 H 20 FNO5 337.1326,found:338.1403; 1 HNMR (400MHz, CDCl3) δ7.88(s,1H),6.71(s,1H),3.95(d,J=5.9Hz,6H),3.72(s,3H),3.67(s,3H),3.34(d dd,J=17.2,8.0,2.4Hz,1H),3.21–3.11(m,1H),2.92(ddd,J=17.2,5.5,2.3Hz,1H),1.29(d,J=7.1Hz,3H).

[0466] 44. Synthesis of M-71 compound

[0467]

[0468] Lithium hydroxide (4.3 mg, 178 umol) was added to a tetrahydrofuran (1 mL) / ethanol (0.5 mL) / water (0.5 mL) solution containing M-69 (6.0 mg, 17.8 umol) prepared in Part 43 of this Example, and stirred at 60°C for 3 hours. The reaction solution was quenched with dilute hydrochloric acid, the pH was adjusted to 3, and then extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by silica gel column to obtain M-71 (4.8 mg, 14.8 umol, 83% yield).

[0469] M-71: 1H NMR (600MHz, CDCl3) δ7.32(s,1H),6.52(s,1H),4.00(s,3H),3.96(s,3H),3.94(s,3H),3.44(d d,J=16.8,7.9Hz,1H),3.17–3.10(m,1H),3.07(dd,J=16.9,5.2Hz,1H),1.32(d,J=7.0Hz,3H); 13 C NMR (151MHz, CDCl3) δ190.53,181.57,154.41,154.38,150.33,148.68,137.23,137.15,133.53,132. 32,132.24,110.38,110.24,108.04,87.83,87.81,62.04,62.02,56.43,42.42,35.02,32.57,17.05.

[0470] 45. Synthesis of M-72 compound

[0471]

[0472] Lithium hydroxide (4.3 mg, 178 umol) was added to a tetrahydrofuran (1 mL) / ethanol (0.5 mL) / water (0.5 mL) solution containing M-68 (6.0 mg, 17.8 umol) prepared in Part 43 of this Example, and stirred at 60°C for 3 hours. The reaction solution was quenched with dilute hydrochloric acid, the pH was adjusted to 3, and then extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified with a silica gel column to obtain 4-(3-fluoro-5,6-dimethoxy-1-methyl-1H-indol-2-yl)-2-methyl-4-oxobutanoic acid M-72 (4.6 mg, 14.2 umol, 80% yield).

[0473] M-72: 1 H NMR(600MHz,DMSO-d6)δ7.07(s,1H),7.06(s,1H),3.89(s,3H),3.88(s,3H),3.8 1(s,3H),3.24(dd,J=16.8,7.6Hz,1H),2.97-2.86(m,2H),1.18(d,J=5.7Hz,3H); 13C NMR(151MHz,DMSO)δ188.87,177.30,152.11,150.06,148.34,146.79,132.35,118.11,1 17.99,106.56,106.45,98.27,93.32,56.37,56.21,44.59,44.54,34.77,32.60,17.54.

[0474] 46. ​​Synthesis of M-73 compound

[0475]

[0476] Step 1: At zero degrees Celsius and under nitrogen protection, sodium hydrogen (1.23 g, 30.69 mmol, 60% dispersed in mineral oil) was added to a tetrahydrofuran (60 mL) solution of ethyl 5,6-dimethoxyindole-2-carboxylate (5.1 g, 20.46 mmol). After the reaction solution was stirred at this temperature for 1 hour, benzenesulfonyl chloride (7.22 g, 40.9 mmol) was slowly added dropwise to the above reaction solution, and continued to stir at zero degrees Celsius for 1 hour, then heated to room temperature and stirred for 12 hours. The reaction was quenched with water, and the reaction solution was adjusted to neutral with dilute hydrochloric acid, then extracted with ethyl acetate, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and purified using a silica gel column to obtain M-73-int-1 (4.3 g, 11.04 mmol, 85% yield).

[0477] Step 2: 1-Chloromethyl-4-fluoro-1,4-diazabicyclo[2.2.2]octane ditetrafluoroboric acid was added to a solution of M-73-int-1 (1.00 g, 2.57 mmol) in dichloromethane (10 mL) and acetonitrile (10 mL). The reaction solution was stirred at 55 degrees Celsius overnight and the solvent was removed by rotary evaporation under reduced pressure. The residue was extracted with ethyl acetate and water, and the organic phase was dried, filtered, concentrated, and purified by silica gel column to obtain a mixture of F-030 and F-050. Finally, preparative chromatography was used to purify to obtain pure M-73-int-2 (8 mg, 0.020 mmol, 0.76% yield) and M-73-int-3 (7 mg, 0.017 mmol, 0.67% yield), respectively.

[0478] Step 3: Lithium hydroxide (58.9 mg, 2.5 mmol) was added to a solution of M-73-int-2 (100.0 mg, 0.25 mmol) in tetrahydrofuran (10 mL) / ethanol (5 mL) / water (5 mL), and stirred at 60 degrees Celsius for 3 hours. The reaction solution was quenched with dilute hydrochloric acid, the pH was adjusted to 3, and then extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by silica gel column to obtain M-73-int-4 (85.5 mg, 0.22 mmol, 90% yield).

[0479] Step 4: M-73-int-4 (85.5 mg, 0.22 mmol) was dissolved in diphenyl ether (5 mL). The reaction solution was stirred at 260 degrees Celsius for 3 hours and then purified by silica gel column to obtain M-73-int-5 (27.5 mg, 0.14 mmol, 64% yield).

[0480] Step 5: 4-dimethylaminopyridine (1.7 mg, 0.014 mmol), di-tert-butyl dicarbonate (45.8 mg, 0.21 mmol) and triethylamine (58.4 uL, 0.42 mmol) were added to a dichloromethane (10 mL) solution of M-73-int-5 (27.5 mg, 0.14 mmol). The reaction solution was stirred at room temperature for 4 hours, quenched with 10 mL of water, extracted with dichloromethane, and the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column to obtain M-73-int-6 (38.4 mg, 0.13 mmol, 93% yield).

[0481] Step 6: Under nitrogen protection, 2,2,6,6-tetramethylpiperidine (72 mg, 0.51 mmol) was dissolved in 1 mL of anhydrous tetrahydrofuran and cooled to -78 degrees Celsius. n-Butyl lithium (20.4 uL, 0.51 mmol, 2.5 M in n-hexane) was added dropwise to the above reaction flask and stirred at this temperature for 1 hour. M-73-int-6 (100 mg, 0.34 mmol) in tetrahydrofuran (2 mL) was added dropwise to the above reaction solution. After the reaction solution was stirred at -78 degrees Celsius for one hour, 2,2-dimethylsuccinic anhydride (87.1 mg, 0.68 mmol) in tetrahydrofuran (1 mL) was added to the above reaction solution and continued to stir at this temperature for 1 hour, then warmed to room temperature and stirred overnight, the reaction was quenched with a small amount of water and hydrochloric acid and the pH was adjusted to 3. The crude product was concentrated and purified using a silica gel column (20:80 = ethyl acetate: petroleum ether) to obtain a mixture of F-034 and F-035 (50.2 mg, 0.12 mmol, 35% yield).

[0482] Step 7: Trifluoroacetic acid (89.1uL, 1.2mmol) was added dropwise to a solution of M-73-int-7 and M-73-int-8 (50.2mg, 0.12mmol) in dichloromethane (2mL), and stirred at room temperature for 2 hours. Then, it was extracted with ethyl acetate and water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column (20:80 = ethyl acetate: petroleum ether) to give a mixture of M-73-int-9 and M-73-int-10 (36.9mg, 0.11mmol, 92% yield).

[0483] Step 8: Diazomethane (165uL, 0.33mmol, 2.0M hexane solution) was added dropwise to a solution of M-73-int-9 and M-73-int-10 (36.9mg, 0.11mmol) in methanol (1mL) and ether (1mL). The reaction solution was stirred at room temperature for 2 hours and then distilled under reduced pressure. The residue was extracted with ethyl acetate and water, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and finally purified using a silica gel column to obtain a mixture of M-73-int-11 and M-73-int-12 (35.3mg, 0.10mmol, 91% yield).

[0484] Step 9: Cesium carbonate (48.9 mg, 0.15 mmol) and iodomethane (18.7 uL, 0.3 mmol) were added to a solution of M-73-int-11 and M-73-int-12 (35.3 mg, 0.10 mmol) in acetonitrile (2 mL). The reaction solution was stirred at 80 degrees Celsius for 12 hours and then the solvent was removed by rotary evaporation under reduced pressure. The residue was extracted with ethyl acetate and water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and then purified using a silica gel column. Purification by preparative chromatography gave M-73 (10 mg, 0.028 mmol, 28% yield) and M-74 (18 mg, 0.051 mmol, 51% yield), respectively.

[0485] M-73: 1 H NMR (600MHz, CDCl3) δ7.15(s,1H),6.55(s,1H),3.96(s,4H),3.93(s,4H),3.88(s,3H),3.60(s,3H),2.88(s,2H),1.52(s,6H).

[0486] 47. Synthesis of M-75 and M-76 compounds

[0487]

[0488] Step 1: Lithium hydroxide (58.9 mg, 2.5 mmol) was added to a tetrahydrofuran (10 mL) / EtOH (5 mL) / water (5 mL) solution containing M-73-int-3 (100.0 mg, 0.25 mmol), and stirred at 60°C for 3 hours, then quenched with dilute hydrochloric acid, and the pH was adjusted to 3. The mixture was extracted with ethyl acetate and water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and finally purified on a silica gel column to obtain M-75-int-1 (85.5 mg, 0.22 mmol, 90% yield).

[0489] Step 2: M-75-int-1 (85.5 mg, 0.22 mmol) was dissolved in diphenyl ether (5 mL). The reaction solution was stirred at 260 °C for 3 hours and then purified using a silica gel column to obtain M-75-int-2 (27.5 mg, 0.14 mmol, 64% yield).

[0490] Step 3: 4-dimethylaminopyridine (1.7 mg, 0.014 mmol), di-tert-butyl dicarbonate (45.8 mg, 0.21 mmol) and triethylamine (58.4 uL, 0.42 mmol) were added to a dichloromethane (10 mL) solution of M-75-int-2 (27.5 mg, 0.14 mmol). The reaction solution was stirred at room temperature for 4 hours and then evaporated to dryness, and extracted with water and ethyl acetate. The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and finally purified on a silica gel column to obtain M-75-int-3 (38.4 mg, 0.13 mmol, 93% yield).

[0491] Step 4: Under nitrogen protection, 2,2,6,6-tetramethylpiperidine (72 mg, 0.51 mmol) was dissolved in 1 mL of anhydrous tetrahydrofuran and cooled to -78 degrees Celsius. n-Butyl lithium (20.4 uL, 0.51 mmol, 2.5 M in n-hexane) was added dropwise to the above reaction flask and stirred at this temperature for 1 hour. A tetrahydrofuran solution (2 mL) of M-75-int-3 (100 mg, 0.34 mmol) was added dropwise to the above reaction solution. After the reaction solution was stirred at -78 degrees Celsius for one hour, a tetrahydrofuran solution (1 mL) of 2,2-dimethylsuccinic anhydride (87.1 mg, 0.68 mmol) was added to the above reaction solution and continued to stir at this temperature for 1 hour, then warmed to room temperature and stirred overnight, the reaction was quenched with a small amount of water and hydrochloric acid and the pH was adjusted to 3. The crude product was concentrated and purified using a silica gel column (20:80 = ethyl acetate: petroleum ether) to give a mixture of M-75-int-4 and M-75-int-5 (50.2 mg, 0.12 mmol, 35% yield).

[0492] Step 5: Trifluoroacetic acid (89.1uL, 1.2mmol) was added dropwise to a solution of M-75-int-4 and M-75-int-5 (50.2mg, 0.12mmol) in dichloromethane (2mL), and stirred at room temperature for 2 hours. Then, it was extracted with ethyl acetate and water. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column (20:80 = ethyl acetate: petroleum ether) to give M-75-int-6 and M-75-int-7 (36.9mg, 0.11mmol, 92% yield).

[0493] Step 6: Diazomethane (165uL, 0.33mmol, 2.0M hexane solution) was added dropwise to a solution of M-75-int-6 and M-75-int-7 (36.9mg, 0.11mmol) in methanol (1mL) and ether (1mL). The reaction solution was stirred at room temperature for 2 hours and then distilled under reduced pressure. The residue was extracted with ethyl acetate and water, the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, and finally purified using a silica gel column to obtain M-75-int-8 and M-75-int-9 (35.3mg, 0.10mmol, 91% yield).

[0494] Step 7: Cesium carbonate (48.9 mg, 0.15 mmol) and iodomethane (18.7 uL, 0.3 mmol) were added to a solution of M-75-int-8 and M-75-int-9 (35.3 mg, 0.10 mmol) in acetonitrile (2 mL). The reaction solution was stirred at 80 ° C for 12 hours and then the solvent was removed by rotary evaporation under reduced pressure. The residue was extracted with ethyl acetate and water, and the organic phase was dried over anhydrous sodium sulfate, filtered, concentrated, purified by silica gel column, and then purified by preparative chromatography to obtain M-75 (10 mg, 0.028 mmol, 28% yield) and M-76 (18 mg, 0.051 mmol, 51% yield), respectively.

[0495] M-75: 1 H NMR(600MHz, CDCl3) δ6.99(d,J=1.8Hz,1H),6.82(d,J=0.8Hz,1H),4.06(s,3H),3.97

[0496] (s,3H),3.89(s,3H),3.60(s,3H),2.86(s,2H),1.49(s,6H); 13C NMR (151MHz, CDCl3) δ198.66,171.91,149.16,149.14,144.87,136.46,135.09,123.71,123.46,123. 41,122.53,122.49,115.94,111.39,108.22,97.96,62.09,56.40,51.57,45.98,45.63,34.56,26.79.

[0497] M-76: 1 H NMR (600MHz, CDCl3) δ7.13(d,J=1.7Hz,1H),6.81(d,J=1.1Hz,1H),4.21(d,J=1.0

[0498] Hz,3H),3.98(d,J=0.4Hz,3H),3.90(s,3H),3.70(s,3H),3.25(s,2H),1.32(s,6H); 13 C NMR (151MHz, CDCl3) δ190.85,177.86,149.33,149.32,144.86,144.79,143.16,137.27,137.19,137.12,1 35.52,124.77,124.72,122.47,122.43,111.41,98.06,62.04,56.38,52.01,49.61,40.42,34.76,25.73.

[0499] 48. Synthesis of M-77 compound

[0500]

[0501] Lithium hydroxide (4.1 mg, 171 umol) was added to a solution of M-75 (6.0 mg, 17.1 umol) prepared in part 50 of this example in tetrahydrofuran (1 mL) / ethanol (0.5 mL) / water (0.5 mL), and stirred at 60°C for 3 hours. The reaction solution was quenched with dilute hydrochloric acid, the pH was adjusted to 3, and then extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by silica gel column to obtain M-77 (4.8 mg, 14.2 umol, 83% yield).

[0502] M-77: 1H NMR (600MHz, CDCl3) δ6.99(s,1H),6.81(s,1H),4.01(s,3H),3.97(s,3H),3.89(s,3H),2.88(s,2H),1.50(s,6H); 13 C NMR (151MHz, CDCl3) δ198.56,176.60,149.22,144.84,143.21,136.59,136.51,134.79,123.58, 123.53,122.49,122.45,108.53,97.99,62.08,62.06,56.40,45.87,45.46,34.56,34.51,26.85.

[0503] 49. Synthesis of M-78 compound

[0504]

[0505] Lithium hydroxide (4.1 mg, 171 umol) was added to a solution of M-76 (6.0 mg, 17.1 umol) prepared in part 50 of this example in tetrahydrofuran (1 mL) / ethanol (0.5 mL) / water (0.5 mL), and stirred at 60°C for 3 hours. The reaction solution was quenched with dilute hydrochloric acid, the pH was adjusted to 3, and then extracted with ethyl acetate, the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated, and the residue was purified by silica gel column to obtain M-78 (4.2 mg, 12.5 umol, 73% yield).

[0506] M-78: 1 H NMR (600MHz, CDCl3) δ7.15(d,J=1.5Hz,1H),6.81(d,J=0.5Hz,1H),4.22(s,3H),3.98

[0507] (s,3H),3.90(s,3H),3.28(s,2H),1.36(s,6H); 13 C NMR (151MHz, CDCl3) δ190.87,181.33,149.37,144.77,143.13,137.32,135.31,129.93,129.90,124.92, 124.87,122.46,122.42,111.69,98.07,62.07,62.05,56.37,49.21,40.16,34.86,34.80,29.33,25.66.

[0508] 50. Synthesis of M-79 compound

[0509]

[0510] Step 1: At 0 ° C, (S)-2,3-dihydroxypropionic acid methyl ester (500.0 mg, 4.16 mmol) and imidazole (283.4 mg, 4.16 mmol) were dissolved in DCM (5.0 mL) and tert-butyldimethylsilyl chloride (342.8 mg, 4.16 mmol) was added, and then stirred at room temperature for 1 hour. After the reaction was completed, the mixture was diluted with water and extracted with ethyl acetate, and then washed with brine. It was then dried over anhydrous sodium sulfate, and the organic phase was concentrated under reduced pressure and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10: 1) to give M-79-int-1 (500.0 mg, 2.13 mmol, yield 51.2%).

[0511] Step 2: At 0 ° C, M-79-int-1 (400.0 mg, 1.71 mmol) was dissolved in dichloromethane (8.0 mL) and Dess-Martin periodinane (868.7 mg, 2.05 mmol) was added, and the reaction mixture was stirred at room temperature for 16 hours. TLC monitored the reaction progress. After the reaction was completed, the mixture was diluted with water and extracted with ethyl acetate, and then washed with brine. The organic layer was then dried over anhydrous Na2SO4 and concentrated under reduced pressure, and the compound 3-(tert-butyl(dimethyl)silyl)oxy-2-oxopropanoic acid methyl ester M-79-int-2 (400.0 mg, 1.72 mmol, yield 100%) was obtained by purification by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1).

[0512] Step 3: At 0 ° C, M-79-int-2 (300.0 mg, 0.87 mmol) was dissolved in tetrahydrofuran (6.0 mL) and sodium hydride (70.3 mg, 1.76 mmol, 60% in paraffin oil) was added. Under argon protection, the mixture was stirred at 0 ° C for 30 minutes. Then M-79-int-2 (246.3 mg, 1.76 mmol) was added dropwise at 0 ° C. Under argon protection, the mixture was stirred at room temperature for 1 h. The reaction mixture was then quenched with saturated ammonium chloride (20.0 mL), diluted with water (20.0 mL), and extracted with ethyl acetate (50.0 mL). The organic phases were combined and dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. Purification by silica gel column chromatography (petroleum ether: ethyl acetate = 10: 1) gave a yellow solid M-79-int-3 (260.0 mg, 0.56 mmol, 66.1% yield).

[0513] M-79-int-3:MS m / z(ESI):448.4[M+H] + .

[0514] Step 4: M-79-int-3 (200.0 mg, 0.43 mmol) and Pd / C (52.6 mg, 10%) were dissolved in methanol (4.0 mL) and stirred at room temperature for 4 hours under a hydrogen balloon. The reaction was then detected by LCMS. The reaction solution was filtered and the filtrate was concentrated in vacuo. The crude product was purified by silica gel column chromatography (PE: EA = 20: 1) to give M-79-int-4 (140.0 mg, 0.30 mmol, yield 70.0%) as a white solid.

[0515] M-79-int-4:MS m / z(ESI):450.5[M+H] + .

[0516] Step 5: SG-111-7 (140.0 mg, 0.31 mmol) was dissolved in tetrahydrofuran (3.0 mL) at 0°C and tetrabutylammonium fluoride (162.8 mg, 0.62 mmol) was added. The reaction was then stirred at room temperature for 16 hours. Saturated ammonium chloride (5.0 mL) was added, diluted with water, the mixture was extracted with ethyl acetate (5.0 mL×3), the combined organic layer was washed with brine and dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate=3 / 1) to give M-79 (29.4 mg, 87.52 umol, yield 28.8%).

[0517] M-79:MS m / z(ESI):336.3[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.36(s,1H),7.13(s,1H),7.05(s,1H),4.93(t,J=5.4Hz,1H),3.95(s,3H),3.8 7(s,3H),3.78(s,3H),3.64(t,J=5.6Hz,2H),3.34-3.39(m,1H),3.06-3.17(m,1H),δ3.00-3.05(m,1H).

[0518] Example 2: Preparation of dimer compound

[0519] 1. Synthesis of compounds D-1 and D-2

[0520]

[0521] Step 1: To a solution of M-29 (67.00 mg, 219.44 umol) prepared in step 18 of Example 1 in acetonitrile (5.0 mL) was added 1,3-dibromopropane (88.60 mg, 438.88 umol) and potassium carbonate (151.64 mg, 1.10 mmol). The mixture was stirred at 70 ° C for 16 hours. The solvent was removed in vacuo and purified with a preparative plate (PE / EA=3 / 1) to give D-1-int-1 (48.00 mg, 112.60 umol, yield 51.31%).

[0522] D-1-int-1:MS m / z(ESI):426.1,428.1[M+H] + .

[0523] Step 2: Potassium carbonate (46.68 mg, 337.79 umol) and M-30 (25.0 mg, 81.88 umol) prepared in part of step 18 of Example 1 were added to a solution of D-1-int-1 (48.00 mg, 112.60 umol) in DMF (1.0 mL). The reactants were stirred at 80 ° C for 16 hours. The reaction mixture was concentrated in high vacuum and purified by preparation plate (DCM / MeOH=20 / 1) to give a white solid D-1 (70.00 mg, 107.57 umol, 95.54% yield).

[0524] D-1:MS m / z(ESI):651.1[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.38(s,1H),7.36(s,1H),7.18(s,1H),7.13(s,1H),7.1 1(s,1H),7.06(s,1H),4.27(t,J=6.2Hz,2H),4.17(t,J=6.2Hz,2H),3.94(d,J=5 .6Hz,6H),3.86(s,3H),3.79(s,3H),3.59(s,6H),3.31-3.26(m,2H),3.08(dd,J =17.2,5.6Hz,2H),2.99-2.89(m,2H),2.30-2.23(m,2H),1.17(d,J=7.1Hz,6H).

[0525] Step 3: Lithium hydroxide monohydrate (8.25 mg, 196.71 umol) was added to a solution of D-1 (32.00 mg, 49.18 umol) in methanol (2.0 mL) and water (1.0 mL). The mixture was stirred at 25 ° C for 16 hours. The pH of the mixture was adjusted to 3 with dilute hydrochloric acid (1N), and purified by perp-HPLC to obtain D-2 (5.00 mg, 8.03 umol, 16.33% yield).

[0526] D-2:MS m / z(ESI):623.2[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ12.11(br.s,2H),7.37(s,1H),7.34(s,1H),7.18(s,1H), 7.13(s,2H),7.11(s,2H),7.06(s,1H),4.27(t,J=6.2Hz,2H),4.17(t,J=6.2Hz,2 H),3.95(d,J=5.9Hz,6H),3.86(s,3H),3.79(s,3H),3.30-3.24(m,2H),2.98(dd, J=16.8,5.6Hz,2H),2.92-2.82(m,2H),2.30-2.22(m,2H),1.15(d,J=7.2Hz,6H).

[0527] 2. Synthesis of D-3 compound

[0528]

[0529] Step 1: Potassium carbonate (34.0 mg, 0.25 mmol) and 1,3-dibromopropane (33.1 mg, 0.16 mmol) were added to a solution of M-30 (25.0 mg, 81.88 umol) prepared in step 18 of Example 1 in acetonitrile (1.0 mL). The reaction was stirred at 70 ° C for 16 hours. The reaction mixture was filtered and concentrated to give a crude product D-3-int-1 (35.0 mg, 81.88 umol, 100% yield).

[0530] D-3-int-1:MS m / z(ESI):426.2,428.2[M+H] + .

[0531] Step 2: Potassium carbonate (34.0 mg, 0.25 mmol) and M-30 (25.0 mg, 81.88 umol) were added to a DMF (1.0 mL) solution of D-3-int-1 (34.9 mg, 81.88 umol). The reactants were stirred at 80 ° C for 16 hours. The reaction mixture was concentrated in high vacuum and purified by a preparative plate (DCM / MeOH=20 / 1) to give a white solid D-3 (18.0 mg, 27.66 umol, two-step yield of 33.8%).

[0532] D-3:MS m / z(ESI):651.3[M+H] + ; 1 H NMR (400MHz, CDCl3) δ7.20 (s, 2H), 7.03 (s, 2H), 6.81 (s, 2H), 4.39 (t, J = 6.0Hz, 4H), 3.96 (s, 6H), 3.89 (s,6H),3.70(s,6H),3.37-3.43(m,2H),2.98-3.13(m,4H),2.49-2.52(m,2H),1.27(d,J=7.2Hz,6H).

[0533] 3. Synthesis of compounds D-4 and D-5

[0534]

[0535] Step 1: K2CO3 (339.5 mg, 2.46 mmol) ...) (339.5 mg, 2.46 mmol) (339.5 mg, 2.46 mmol)) (339.5 mg, 2.46 mmol) (339.5 mg, 2.46 mmol)) (339.5 mg, 2.46 mmol) (339.5 mg, 2.46 mmol)) (339.5 mg, 2.46 mmol) (339.5 mg, 2.46 mmol)) (339.5 mg, 2.46 mmol) (339.5 mg, 2.46 mmol)) (339.5 mg, 2.46 mmol) (339.5 mg, 2.46 mmol))

[0536] D-4-int-1:MS m / z(ESI):426.1[M+H] + .

[0537] Step 2: M-29 (71.6 mg, 234.58 umol) and K2CO3 (97.3 mg, 703.73 umol) were added to a solution of D-4-int-1 (100.0 mg, 234.58 umol) in DMF (2.0 mL). The mixture was stirred at 70 ° C for 16 hours. The mixture was filtered and the filtrate was concentrated. The residue was purified by preparative TLC (petroleum ether: ethyl acetate = 1: 1) to give D-4 (67.0 mg, 0.12 mmol, 42.1% yield) as an off-white solid.

[0538] D-4:MS m / z(ESI):673.2[M+Na] + ; 1 H NMR (400MHz, DMSO-d6) δ7.36(s,2H),7.17(s,2H),7.06(s,2H),4.16(t,J=6.2Hz,4H),3.95(s,6H),3.87(s,6H),3.59(s,6H),3.33 –3.27(m,2H),3.10(d,J=5.4Hz,1H),3.06(d,J=5.4Hz,1H),2.99–2.91(m,2H),2.23(dd,J=12.0,6.0Hz,2H),1.17(d,J=7.2Hz,6H).

[0539] Step 3: LiOHH2O (9.7 mg, 230.52 umol) was added to a mixed solution of THF (1.0 mL), MeOH (0.5 mL), H2O (0.5 mL) and DMSO (0.5 mL) containing D-4 (30.0 mg, 46.10 umol). The mixture was stirred at room temperature for 16 hours, and the mixture was concentrated to remove the organic layer, and the residue was extracted with EtOAc (5.0 mL x 2) and water (10.0 mL). The pH value of the aqueous layer was adjusted to 1-2 with HCl (1 N, aq). Filter and dry the filter cake to obtain D-5 (24.9 mg, 40.09 umol, 86.95% yield) as a white solid.

[0540] D-5:MS m / z(ESI):623.3[M+H] + ; 1H NMR (400MHz, DMSO-d6) δ12.05(s,2H),7.35(s,2H),7.18(s,2H),7.06(s,2H),4.16(t,J=6.2Hz,4H),3.96(s,6H),3.87(s,6H),3.27 (d,J=8.2Hz,2H),2.99(dd,J=16.8,5.6Hz,2H),2.87(dd,J=13.2,7.6Hz,2H),2.54(s,4H),2.27-2.17(m,2H),1.16(d,J=7.2Hz,6H).

[0541] 4. Synthesis of D-6 and D-7 compounds

[0542]

[0543] Step 1: At -78 ° C, LiHMDS (1M in THF, 0.88 mL, 0.88 mmol) was added to a THF (10.0 mL) solution containing tert-butyl acetate (102.7 mg, 0.88 mmol), and the mixture was stirred at -78 ° C for 1 hour, and then 5-(benzyloxy)-6-methoxy-1-methyl-1H-indole-2-carboxylic acid ethyl ester (dissolved in 2.0 mL THF, 100.0 mg, 0.29 mmol, 1.00 equivalent) was added dropwise. The reaction mixture was stirred at room temperature for 2 hours. The mixture was carefully treated with aqueous ammonium chloride (50.0 mL). The mixture was extracted with ethyl acetate (20.0 mL × 3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by preparative plate (EA / PE=1 / 3) to obtain D-6-int-1 (120.0 mg, 0.29 mmol, 99.3% yield).

[0544] D-6-int-1:MS m / z(ESI):354.1[M-56+H] + .

[0545] Step 2: Potassium tert-butoxide (164.4 mg, 1.47 mmol) and hexadecyltrimethylammonium bromide (44.5 mg, 0.12 mmol) were added to a solution of tert-butanol (20.0 mL) containing D-6-int-1 (500.0 mg, 1.22 mmol), and the mixture was heated to 80 ° C. Ethyl acrylate (122.3 mg, 1.22 mmol) was then added. The reaction mixture was stirred at 80 ° C for 0.5 hours. The mixture was treated with aqueous ammonium chloride (20.0 mL), extracted with ethyl acetate (20.0 mL×3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column chromatography (EA / PE=1 / 8) to give crude D-6-int-2 (600.0 mg, 1.18 mmol, 96.4% yield, purity 95%).

[0546] D-6-int-2:MS m / z(ESI):454.3[M-56+H] + .

[0547] Step 3: trifluoroacetic acid (1.50 mL, 19.62 mmol) was added to a DCM (5.0 mL) solution containing D-6-int-2 (500.0 mg, 0.98 mmol), and then stirred at room temperature for 16 hours. The reaction mixture was concentrated to give the crude product D-6-int-3 (445.0 mg, 0.98 mmol) without purification.

[0548] D-6-int-3:MS m / z(ESI):454.2[M+H] + .

[0549] Step 4: D-6-int-3 (445 mg, 0.98 mmol) was dissolved in toluene (10.0 mL), heated to 80° C., and stirred for 3 hours. The reaction mixture was concentrated in high vacuum and purified by high performance liquid chromatography (Column: Waters Xbridge 20*150mm 10um, C18, Mobile Phase A: water, B: ACN, 0.5% FA) to give D-6-int-4 (210.0 mg, 0.51 mmol, two-step yield 52.3%).

[0550] D-6-int-4:MS m / z(ESI):410.3[M+H] + ; 1H NMR (400MHz, CDCl3) δ7.46-7.48(m,2H), δ7.31-7.39(m,2H), δ7.15(s,1H),7.06(s,1H),6.75(s,1H),5.17(s,2H),4.11-4. 16(m,2H),4.04(s,3H),3.99(s,3H),2.96(t,J=7.4Hz,2H),2.41(t,J=7.2Hz,2H),2.05-2.10(m,2H),1.25(t,J=7.0Hz,3H).

[0551] Step 5: Pd / C (10% carbon, about 55% water, 16.0 mg, mass ratio 10%) was added to a solution of D-6-int-4 (160.0 mg, 0.39 mmol) in ethanol (5.0 mL). The mixture was stirred at room temperature under hydrogen for 1 hour. The mixture was filtered and the filtrate was concentrated and purified by silica gel column chromatography (PE: EA = 3: 1) to give a white solid D-6-int-5 (110 mg, 0.34 mmol, 88.2% yield).

[0552] D-6-int-5:MS m / z(ESI):320.1[M+H]+; 1 H NMR(400MHz,DMSO-d6)δ8.70(s,1H),δ7.25(s,1H),7.00(s,1H),6.96(s,1H),4.03-4.08(m,2H),3.95(s,3H),3.88 (s,3H),3.42-3.44(m,1H),2.92(t,J=7.2Hz,2H),2.37(t,J=7.4Hz,2H),1.85-1.89(m,2H),1.17(t,J=7.2Hz,3H).

[0553] Step 6: D-6-int-5 (30.0 mg, 93.94 umol) was dissolved in acetonitrile (1.0 mL), and potassium carbonate (39.0 mg, 281.82 umol) and 1,3-dibromopropane (38.0 mg, 187.88 umol) were added. The reaction was stirred at 70 ° C for 16 hours. The reaction mixture was filtered and concentrated to obtain the crude product D-6-int-6 (34.5 mg, 78.28 umol) without purification.

[0554] D-6-int-6:MS m / z(ESI):440.1,442.1[M+H] + .

[0555] Step 7: D-6-int-6 (34.5 mg, 78.28 umol) was dissolved in DMF (1.0 mL), and potassium carbonate (34.0 mg, 234.85 umol) and D-6-int-5 (25.0 mg, 7828 umol) were added. The reactant was stirred at 80 ° C for 16 hours. The reaction mixture was concentrated under high vacuum and purified by thin layer preparative chromatography (dichloromethane / methanol=20 / 1) to give a white solid D-6 (25.0 mg, 36.83 umol, 47.1% yield).

[0556] D-6:MS m / z(ESI):679.2[M+H] + ,1H NMR (400MHz, CDCl3) δ7.17(s,2H),7.12(s,2H),6.73(s,2H),4.29(t,J=6.2Hz,4H),4.14(q,J=7.2Hz,4H),4.0 4(s,6H),3.95(s,6H),2.97(t,J=7.2Hz,4H),2.43(t,J=7.4Hz,6H),2.04-2.11(m,4H),1.26(t,J=7.2Hz,6H).

[0557] Step 8: D-6 (20.0 mg, 29.47 umol) was dissolved in a mixture of methanol (2.0 mL) and water (1.0 mL), and lithium hydroxide monohydrate (5.0 mg, 0.13 mmol) was added, and then stirred at room temperature for 16 hours. The reaction mixture was adjusted to pH <4 with dilute hydrochloric acid, and purified by high performance liquid preparative chromatography (A: 0.1% TFA aqueous solution, B: acetonitrile) to obtain a white solid D-7 (8.0 mg, 0.0128 mmol, yield 43.6%).

[0558] D-7:MS m / z(ESI):623.3[M+H] + ; 1 H NMR (400MHz, CDCl3) δ12.06(br.s,2H),7.31(s,2H),7.17(s,2H),7.06(s,2H),4.15(t,J=6.0Hz,4H),3.9 7(s,6H),3.87(s,6H),2.94(t,J=7.4Hz,4H),2.31(t,J=7.4Hz,4H),2.18-2.25(m,2H),1.81-1.88(m,4H).

[0559] 5. Synthesis of D-8 compound

[0560]

[0561] D-6-int-5 (30.0 mg, 93.94 1.94 umol) prepared in step 52 of Example 1 was dissolved in acetonitrile (1.0 mL), and potassium carbonate (39.0 mg, 281.82 umol) and D-4-int-1 (40.1 mg, 93.94 umol) were added, and the reaction was stirred at 80° C. for 16 hours. The reaction mixture was filtered and concentrated under high vacuum, and purified by thin layer preparative chromatography (dichloromethane / methanol=20 / 1) to give a white solid D-8 (25.0 mg, 37.61 umol, 40.0% yield).

[0562] D-8:MS m / z(ESI):665.4[M+H] + ; 1 H NMR (400MHz, CDCl3) δ7.18 (d, J=4.8Hz, 2H), 7.12 (d, J=

[0563] 0.8Hz,2H),6.72(d,J=3.6Hz,2H),4.29(t,J=6.2Hz,4H),4.14(q,J=7.2Hz,2H),4.02(d,J=7.6Hz,6H),3.94(d,J=1.4Hz,6H),3.71( s,3H),3.41(dd,J=16.4,7.6Hz,1H),3.07-3.16(m,1H),2.93-3.06(m,3H),2.39-2.47(m,4H),2.02-2.14(m,2H),1.22-1.30(m,6H).

[0564] 6. Synthesis of D-9 compound

[0565]

[0566] Step 1: M-29 (200.0 mg, 0.66 mmol) prepared in part of step 18 of Example 1 was dissolved in acetonitrile (5.0 mL) at room temperature, and potassium carbonate (452.7 mg, 3.28 mmol) and 1,4-dibromobutane (707.2 mg, 3.28 mmol) were added, and the reaction mixture was stirred at 80 ° C for 16 hours. After the reaction was completed, the mixture was diluted with water (20.0 mL) and extracted with ethyl acetate (20.0 mL×3). It was then dried over anhydrous sodium sulfate and concentrated under reduced pressure, and purified by silica gel column chromatography to obtain D-9-int-1 (70.0 mg, 0.16 mmol, yield 5.2%) as a yellow oil.

[0567] D-9-int-1:MS m / z(ESI):440.3,442.3[M+H]+ .

[0568] Step 2: D-9-int-1 (70.0 mg, 0.16 mmol) was dissolved in acetonitrile (5.0 mL) solution at room temperature and potassium carbonate (109.85 mg, 0.79 mmol, 5.00 eq) and M-29 (48.54 mg, 0.16 mmol) prepared in step 18 of Example 1 were added, and the reaction mixture was stirred at 80 ° C for 16 hours. After the reaction was completed, the reaction solution was diluted with water (10.0 mL) and ethyl acetate (10.0 mL × 3). It was then dried over anhydrous sodium sulfate and concentrated under reduced pressure, and purified by silica gel column chromatography (petroleum ether: ethyl acetate = 1: 1) to obtain D-9 (15.0 mg, 0.02 mmol, yield 14.2%) as a white solid.

[0569] D-9:MS m / z(ESI):665.6[M+H] + ; 1 H NMR (400MHz, CDCl3) δ7.19(s,2H),7.07(s,2H),6.71(s,2H),4.14–4.17(m,4H),4.02(s,6H),3.94(s,6H),3.71(s,6H),3.41( dd,J=16.4,7.6Hz,2H),3.12(dd,J=13.6,6.4Hz,2H),3.01(dd,J=16.4,6.0Hz,2H),2.10-2.13(m,4H),1.27(d,J=6.8Hz,6H).

[0570] 7. Synthesis of D-10 and D-11 compounds

[0571]

[0572] Step 1: M-29 (1.0 g, 3.28 mmol) and 2-(chloromethyl)oxirane (1.52 g, 16.38 mmol) prepared in part in step 18 of Example 1 were dissolved in acetone (15.0 mL), followed by the addition of potassium carbonate (905.3 mg, 6.55 mmol) and tetrabutylammonium bromide (527.91 mg, 1.64 mmol). The mixture was stirred at 60 ° C for 16 hours. The mixture was diluted with water (50.0 mL) and extracted with ethyl acetate (50.0 mL×3). The organic layer was washed with brine, dried over anhydrous Na2SO4 and concentrated. The residue was purified by silica gel column (petroleum ether / ethyl acetate=10:1) to give D-10-int-1 (802.0 mg, 2.22 mmol, 67.76% yield) as a white solid.

[0573] D-10-int-1:MS m / z(ESI):362.3[M+H] + .

[0574] Step 2: D-10-int-1 (850.0 mg, 2.35 mmol) was dissolved in a dry N-methylpyrrolidone (15.0 mL) solution, and M-29 (718.14 mg, 2.35 mmol) and potassium carbonate (650.13 mg, 4.70 mmol) prepared in step 18 of Example 1 were added. The reaction was stirred at 150 ° C for 2 hours using microwave heating. The mixture was filtered and concentrated, and purified by HPLC (C18, 0.1% formic acid in water, acetonitrile) to give a white solid D-10 (760.0 mg, 1.14 mmol, 48.47% yield).

[0575] D-10:MS m / z(ESI):667.6[M+H] + .

[0576] Step 3: D-10 (760.0 mg, 1.14 mmol) was dissolved in dichloromethane (15.0 mL) and triethylamine (346.1 mg, 3.42 mmol) and methanesulfonyl chloride (195.87 mg, 1.71 mmol) were added. The mixture was stirred at room temperature for 3 hours. LCMS found the desired MS. The mixture was treated with aqueous ammonium chloride and extracted with dichloromethane, dried over anhydrous sodium sulfate, filtered and concentrated, and purified by silica gel column (dichloromethane / methanol=30 / 1) to give an off-white solid D-11-int-1 (624.0 mg, 837.80 umol, 73.50% yield).

[0577] D-11-int-1:MS m / z(ESI):745.6[M+H] + .

[0578] Step 4: D-11-int-1 (620.0 mg, 832.43 umol) was dissolved in a solution of N, N-dimethylformamide (10.0 mL) and sodium azide (432.9 mg, 6.66 mmol) was added. The mixture was stirred at 80 ° C for 48 hours. The mixture was treated with water (50.0 mL) and extracted with ethyl acetate (20.0 mL×3), dried over anhydrous sodium sulfate, filtered, concentrated and purified by silica gel column (dichloromethane / methanol=30 / 1) to obtain an off-white solid D-11-int-2 (440.0 mg, 636.09 umol, yield 76.41%).

[0579] D-11-int-2:MS m / z(ESI):692.4[M+H] +.

[0580] Step 5: Pd / C (45.0 mg, 10% C, moistened with about 55% water) was added to a solution of D-11-int-2 (440 mg, 636.09 umol) dissolved in tetrahydrofuran (10.0 mL). The mixture was stirred at room temperature under a hydrogen balloon for 6 hours. The mixture was filtered and concentrated. The residue was purified by silica gel column (dichloromethane / methanol=10 / 1) to give D-11 (350.0 mg, 525.74 umol, yield 82.65%) as a white solid.

[0581] D-11:MS m / z(ESI):666.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.35(s,2H),7.18(s,2H),7.06(s,2H),4.06(dd,J=9.6,5.6Hz,2H),3.99(t,J=4.8Hz,2H),3.95(s,6H),3.8 7(s,6H),3.59(s,6H),3.48-3.41(m,1H),3.31-3.28(m,2H),3.08(dd,J=17.2,5.2Hz,2H),3.01-2.90(m,2H),1.17(d,J=7.2Hz,6H).

[0582] 8. Synthesis of D-12 compound

[0583]

[0584] Step 1: D-11 (73.6 mg, 0.11 mmol) prepared in part 7 of this example was dissolved in 1.0 mL of tetrahydrofuran, Ac2O (33.8 mg, 0.33 mmol) and DIPEA (21.4 mg, 0.17 mmol) were added to the solution, and the mixture was reacted at 25°C for 2 hours. After evaporation of the solvent, the crude product was purified by flash column chromatography to obtain a white product D-12-int-1 (61.5 mg, 86.9 umol) with a yield of 79%.

[0585] Step 2: D-12-int-1 (28.4 mg, 40 umol) was dissolved in 0.5 mL DMF, 0.4 mL tert-butyl alcohol and 0.2 mL water were added to the solution, and then lithium hydroxide (2.9 mg, 0.12 mmol) was added. The reaction was stirred at 25 ° C for 6 hours. After the reaction was completed, 0.12 mL HCl (1.0 N) was added to neutralize the reaction solution to pH 7.0. 3 mL of water was added, and 5 mL of ethyl acetate was extracted three times. After the organic layer was washed with saturated brine, it was concentrated to obtain D-12 with a yield of 87%.

[0586] D-12:MS m / z(ESI):680.6[M+H] + ; HRMS m / z:680.3183[M+H] + ; 1 H NMR (400 MHz, DMSO-d6)δ

[0587] 8.22(s,1H),8.22(d,J=7.5Hz,1H),7.34(d,J=3.1Hz,2H),7.34(d,J=3.1Hz,2H),7.24(d, J=2.7Hz,2H),7.24(d,J=2.7Hz,2H),7.07(s,2H),7.07(s,2H),4.13(d,J=5.6Hz,4H),3.9 6(s,6H),3.86(s,6H),3.41–3.21(m,22H),3.07–2.93(m,2H),2.88(dd,J=8.1,4.8Hz,2H) ,2.50(dt,J=3.6,1.8Hz,25H),1.90(s,3H),1.24(d,J=8.4Hz,5H),1.16(d,J=7.2Hz,6H).

[0588] 9. Synthesis of D-13 compound

[0589]

[0590] Dissolve D-11 (11.2 mg, 16.8 μmol) prepared in Part 7 of this Example in 0.2 mL of tert-butanol and 0.1 ml of water. Add lithium hydroxide (1.2 mg, 0.05 mmol). Stir the reaction at 25°C for 6 hours. After the reaction is completed, add 0.05 ml of HCl (1.0 N) to neutralize the reaction solution to pH = 7.0. Add 3 ml of water, extract three times with 5 ml of ethyl acetate, wash the organic layer with saturated brine, and concentrate to obtain a white solid D-13 (7.8 mg, 12.2 μmol, yield 73%).

[0591] D-13:MS m / z(ESI):638.56[M+H] + ; HRMS m / z:638.2714[M+H] + ; 1H NMR (400MHz, DMSO-d6) δ12.12(s,2H),8.38(s,3H),7.41(s,2H),7.30(s,2H),7.13(s,2H),4.29(ddd,J=16.3,10.5,5.2Hz,4H),3.97(s,7H),3.89 (s,6H),3.29(d,J=8.4Hz,2H),3.00(dd,J=16.9,5.4Hz,2H),2.93–2.82(m,2H),2.00(dd,J=14.7,7.2Hz,1H),1.23(s,2H),1.17(d,J=7.2Hz,6H).

[0592] 10. Synthesis of D-14 compound

[0593]

[0594] Step 1: Dissolve D-11 (148.1 mg, 0.22 mmol) prepared in part 7 of this example in 1.0 mL of tetrahydrofuran, add Boc2O (72.8 mg, 0.33 mmol), and stir at 25°C for 2 hours. Evaporation and concentration, flash column chromatography, and obtain white solid D-14-int-1 (113 mg, 0.15 mmol) with a yield of 66%.

[0595] Step 2: Dissolve D-14-int-1 (24.5 mg, 32 μmol) in 0.9 mL tert-butanol and 0.3 mL water. Add lithium hydroxide (0.3 mg, 0.1 mmol). Stir and react at 25°C for 6 hours. After the reaction, add 0.05 mL HCl (1.0 N) to neutralize the reaction solution to pH 7.0. Add 3 mL water and extract three times with 5 mL ethyl acetate. Wash the organic layer with saturated brine and concentrate to obtain the crude product D-14-int-2.

[0596] Step 3: Dissolve the crude product D-14-int-2 obtained in step 2 in 1.0 mL DMF, add HATU (18.2 mg, 0.05 mmol), 3-amino-1-propanol (9.6 mg, 0.13 mmol), and DIPEA (16.5 mg, 0.13 mmol), stir at 25 °C for 4 hours, and evaporate and concentrate the reaction solution to obtain the crude product D-14-int-3.

[0597] Step 4: Add 0.6 ml of 5% TFA / DCM to the crude product D-14-int-3 obtained in step 3 and stir for 2 hours. After the reaction is completed, the reaction solution is concentrated and purified by flash column chromatography to obtain a white solid D-14 (13.4 μmol).

[0598] D-14:MS m / z(ESI):752.61[M+H] + ; 1 H NMR(400MHz,DMSO-d6)δ7.84(t,J=5.6Hz,2H),7.35(s,2H),7.25(s,2H),7.09(s,2H ),4.38(s,2H),4.16(ddd,J=27.2,9.9,5.5Hz,4H),3.96(s,6H),3.88(s,6H),3.75–3 .67(m,1H),3.38(dd,J=10.3,6.0Hz,4H),3.25(td,J=9.9,4.9Hz,2H),3.07(q,J=6. 7Hz, 4H), 2.85 (dt, J = 10.9, 6.1Hz, 4H), 1.52 (p, J = 6.6Hz, 4H), 1.08 (d, J = 6.6Hz, 6H).

[0599] 11. Synthesis of compounds D-15, D-16 and D-17

[0600]

[0601] After chiral separation of D-4 prepared in part 3 of this example, chiral pure compounds D-15, D-16 and D-17 were obtained. Chiral separation conditions: System Waters UPCC; Column name DAICEL OJ; Columnsize 100*3mm 3μm; Mobile Phase A Supercritical CO2; Mobile Phase B MeOH(0.1%DEA):ACN=1:1; Wavelength 214nm; Flow 1.5mL / min; Column temp 35℃; Back Pressure (psi) 1800psi.

[0602]

[0603]

[0604] 12. Synthesis of compounds D-18, D-19 and D-20

[0605]

[0606] D-15 (26 mg, 0.04 mmol) prepared in part 10 of this example was dissolved in a mixed solvent of tetrahydrofuran / dimethyl sulfoxide / dichloromethane / methanol / water (0.20 mL / 0.40 mL / 0.20 mL / 0.20 mL / 0.20 mL), and lithium hydroxide (3.8 mg, 0.16 mmol) was added under stirring, and the system was stirred at room temperature for 16 hours. After completion, 1.0 M dilute hydrochloric acid was added to the system, the pH was adjusted to 2-3, and dichloromethane (40 mL×3) was added for extraction, the organic phases were combined, dried over anhydrous sodium sulfate, filtered, rinsed with acetone (10 mL×3), the filtrate was concentrated, the remaining solvent was drained with a high vacuum oil pump, and then the product residue was slurried and purified with n-hexane / dichlorohexane as a solvent to obtain most of the target product; filtered, the mother liquor was concentrated, and further purified by preparative liquid chromatography to obtain a small amount of the target product. The two products were combined and dried in a freeze dryer to obtain an off-white or light yellow solid D-18.

[0607] This reaction is the same as the reaction operation for synthesizing D-18 above, using D-17 prepared in Part 10 of this Example as the raw material, and the reaction yields a light yellow solid D-19.

[0608] This reaction is the same as the reaction operation for synthesizing D-18 above, using D-16 prepared in Part 10 of this Example as the raw material, and the reaction yields a light yellow solid D-20.

[0609]

[0610] Chiral analysis conditions: Column IC-3(IC30CE-PH006); Column size 0.46cmI.D. mg / ml in MeOH50%EtOH 50%; HPLC Shimadzu LC 20A QA&QC-HPLC-12.

[0611] 13. Synthesis of D-21 and D-22 compounds

[0612]

[0613] Step 1: Dissolve 3.1g 5-(benzyloxy)-6-methoxy-1-methyl-1H-indole-2-carboxylic acid (10mmol), 4.6g HATU (12mmol) and 2.6g DIPEA (20mmol) in 310mL THF and react at room temperature for 1h. Add 2.4g 3-(methylamino)propionic acid methyl ester (20mmol) to the reaction solution, heat to 60°C, and react for 12h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 2.5g D-21-int-1.

[0614] Step 2: Dissolve 2.1 g of D-21-int-1 (5.1 mmol) in 40 mL of methanol, add 0.2 g of 10% Pd / C (10% wt.), heat to 50°C, and react under a hydrogen balloon for 12 h. After the reaction is complete, filter, concentrate the filtrate, and perform column chromatography to obtain 1.3 g of D-21-int-2.

[0615] Step 3: Dissolve 640 mg D-21-int-2 (2.0 mmol) and 2.0 g 1,3-dibromopropane (10.0 mmol) in 20 mL acetonitrile, add 828 mg potassium carbonate (6.0 mmol) in batches, raise the temperature to 70°C, and react for 16 hours. After the reaction is complete, filter, concentrate the filtrate, and perform column chromatography to obtain 441 mg D-21-int-3.

[0616] Step 4: Dissolve 440 mg D-21-int-3 (1.0 mmol), 640 mg intermediate D-21-int-3 (2.0 mmol) and 414 mg potassium carbonate (3.0 mmol) in 30.0 mL DMF, heat to 70 ° C, and react for 16 hours. After the reaction is completed, dilute with water, extract with ethyl acetate, combine the organic layers, dry, concentrate, and obtain 347 mg D-21 by column chromatography.

[0617] D-21: HRMS (ESI) m / z: [M+Na] + Calcd for C 35 H 44 N4NaO 10 703.2955;Found703.2954; 1H NMR (600MHz, DMSO-d6) δ7.12(s,2H),7.04(s,2H),6.52(s,2H),4.13(t,J=6.2Hz,4H),3.83(s,6H),3. 76–3.71(m,4H),3.69(s,6H),3.59(s,6H),3.04(s,6H),2.66(t,J=7.1Hz,2H),2.18(p,J=6.3Hz,2H).

[0618] Step 5: 136 mg D-21 (0.2 mmol) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 24 mg LiOH (1.0 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated, and the residue was poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography to obtain 56 mg D-22.

[0619] D-22: HRMS (ESI) m / z: [M+H] + Calcd for C 33 H 41 N4O 10 Found 653.2823; 1 HNMR(600MHz,DMSO-d6)δ7.11(s,2H),7.04(s,2H),6.49(s,2H),4.14(t,J=6.2Hz,4H),3.8 3(s,6H),3.69(s,6H),3.68(s,4H),3.03(s,6H),2.55–2.50(m,4H),2.17(p,J=6.2Hz,1H).

[0620] 14. Synthesis of D-23 and D-24 compounds

[0621]

[0622] Step 1: Dissolve 3.1 g 5-(benzyloxy)-6-methoxy-1-methyl-1H-indole-2-carboxylic acid (10 mmol), 4.6 g HATU (12 mmol) and 5.2 g DIPEA (40 mmol) in 310 mL THF and react at room temperature for 1 h. Add 2.48 g 3-aminopropionic acid methyl ester hydrochloride (20 mmol) to the reaction solution, heat to 60°C, and react for 12 h. After the reaction is completed, cool the reaction solution to room temperature, pour into water, extract with ethyl acetate, combine the organic layers, and wash the organic layers with 1N HCl aqueous solution. Dry the organic layer, concentrate, and separate by column chromatography to obtain 2.8 g D-23-int-1.

[0623] Step 2: Dissolve 2.0 g of intermediate D-23-int-1 (5.1 mmol) in 40 mL of methanol, add 0.2 g of 10% Pd / C (10% wt.), heat to 50°C, and react under a hydrogen balloon for 12 h. After the reaction is complete, filter, concentrate the filtrate, and perform column chromatography to obtain 1.3 g of D-23-int-2.

[0624] Step 3: Dissolve 612 mg of D-23-int-2 (2.0 mmol) and 2.0 g of 1,3-dibromopropane (10.0 mmol) in 20 mL of acetonitrile, add 828 mg of potassium carbonate (6.0 mmol) in batches, raise the temperature to 70°C, and react for 16 hours. After the reaction is complete, filter, concentrate the filtrate, and perform column chromatography to obtain 556 mg of D-23-int-3.

[0625] Step 4: Dissolve 426 mg of intermediate D-23-int-3 (1.0 mmol), 612 mg of intermediate D-23-int-2 (2.0 mmol) and 414 mg of potassium carbonate (3.0 mmol) in 30.0 mL of DMF, heat to 70°C, and react for 16 h. After the reaction is complete, dilute with water, extract with ethyl acetate, combine the organic layers, dry and filter, and obtain 196 mg of D-23 by column chromatography.

[0626] D-23:purity:94%; HRMS(ESI)m / z:[M+H] + Calcd for C 33 H 41 N4O 10 Found 653.2823; 1HNMR(600MHz,DMSO-d6)δ8.37(t,J=5.7Hz,2H),7.13(s,2H),7.03(s,2H),6.91(s,2H),4.13(t,J =6.2Hz,4H),3.92(s,6H),3.82(s,3H),3.60(s,6H),2.58(t,J=7.0Hz,4H),2.18(p,J=6.2Hz,2H).

[0627] Step 5: 130 mg D-23 (0.2 mmol) was dissolved in a mixed solvent of 3.0 mL THF, 3.0 mL MeOH and 1.0 mL H2O, 24 mg LiOH (1.0 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography to obtain 62 mg D-24.

[0628] D-24:purity:97%; HRMS(ESI)m / z:[M+H] + Calcd for C 31 H 37 N4O 10 625.2510;Found 625.2508; 1 HNMR(600MHz,DMSO-d6)δ12.11(s,2H),8.34(t,J=5.6Hz,2H),7.14(s,2H),7.04(s,2H),6.93(s,2 H), 4.14 (t, J = 6.3Hz, 4H), 3.94 (s, 6H), 3.83 (s, 6H), 3.43 (q, J = 6.9Hz, 4H), 2.50 (p, J = 6.3Hz, 2H).

[0629] 15. Synthesis of D-25 and D-26 compounds

[0630]

[0631] Step 1: Dissolve 440 mg of the intermediate D-21-int-3 (1.0 mmol) prepared in step 13 of this example 2, 610 mg of M-29 (2.0 mmol) prepared in step 18 of example 1, and 414 mg of potassium carbonate (3.0 mmol) in 30.0 mL of DMF, heat to 70°C, and react for 16 hours. After the reaction is complete, cool to room temperature, dilute with water, extract with ethyl acetate, combine the organic layers, dry, concentrate, and obtain 228 mg of D-25 by column chromatography.

[0632] D-25:purity:99%; HRMS(ESI)m / z:[M+H] + Calcd for C 35 H 44 N3O 10 Found 666.3027; 1 HNMR(600MHz,DMSO-d6)δ7.36(s,1H),7.17(s,1H),7.13(s,1H),7.05(s,2H),6 .52(s,1H),4.17–4.12(m,4H),3.95(s,3H),3.86(s,3H),3.84(s,3H),3.74(br s,3H),3.70(s,3H),3.60(s,3H),3.59(s,3H),3.34–3.29(m,2H),3.08(dd,J=17.0,5.4Hz,1H), 3.05(s,2H),2.98–2.91(m,1H),2.66(t,J=7.1Hz,2H),2.23–2.17(m,2H),1.17(d,J=7.1Hz,3H).

[0633] Step 2: 133 mg D-25 (0.2 mmol) was dissolved in a mixed solvent of 6.0 mL THF, 6.0 mL MeOH and 2.0 mL H2O, 24 mg LiOH (1.0 mmol) was added in batches, the temperature was raised to 50°C, and the reaction was carried out for 12 h. After the reaction was completed, the reaction solution was cooled to room temperature, concentrated, and the residue was poured into water, the pH was adjusted to 1-2 with 1N HCl aqueous solution, extracted with ethyl acetate, the organic layers were combined, dried, concentrated, and separated by column chromatography to obtain 45 mg D-26.

[0634] D-26:purity:96%;HRMS(ESI)m / z:[M+Na] + Calcd for C 33 H 39 N3NaO 10 660.2533;Found 660.2515; 1HNMR(600MHz,DMSO-d6)δ12.23(s,2H),7.36(s,1H),7.18(s,1H),7.14(s,1H),7.06(s,1H), 7.06(s,1H),6.52(s,1H),4.17–4.12(m,4H),3.97(s,3H),3.87(s,3H),3.84(s,3H),3.71(br s,3H),3.71(s,3H),3.34–3.29(m,1H),3.05(br s,2H),2.99(dd,J=16.8,5.5Hz,1H),2.92–2.85(m,1H),2.58(t,J=7.2Hz,2H),2.21(p,J=6.3Hz,2H),1.17(d,J=7.2Hz,3H).

[0635] 16. Synthesis of D-27 and D-28 compounds

[0636]

[0637] The D-26 prepared in step 15 of this example 2 was subjected to chiral column separation to obtain D-27 (ee value 98%) and D-28 (ee value 98%). Separation conditions: Column: CHIRALCEL OZ-H (OZHOCE-BO005); Column size: 0.46 cm I.D. × 25 cm L; Mobile phase: MeOH / FAc = 100 / 0.1 (V / V); Flow rate: 1.0 ml / min; Wavelength: UV 365 nm; Temperature: 35°C; Shimadzu LC-20AT CP-HPLC-09.

[0638] 17. Synthesis of D-29 compound

[0639]

[0640] D-5 (17.2 mg, 27.6 μmol) prepared in part 3 of this example was dissolved in 1.0 mL of DMF, HATU (42 mg, 0.11 mmol), 3-amino-1-propanol (8.3 mg, 0.11 mmol) and DIPEA (17.8 mg, 0.14 mmol) were added to the solution, and the mixture was reacted at 25° C. for 4 hours. After the reaction solution was concentrated, the crude product was purified by flash column chromatography to obtain white solid D-29 (9.4 mg, 12.8 μmol, 47% yield).

[0641] D-29:MS m / z(ESI):737.67[M+H]+ ; 1 H NMR (400MHz, DMSO-d6) δ7.83 (t, J=5.6Hz, 2H), 7.32 (s,

[0642] 2H),7.18(s,2H),7.05(s,2H),4.38(t,J=5.3Hz,2H),4.16(t,J=6.2Hz,4H),3.95(s,6H),3.87(s,6H),3.38(dd,J=11.6,6.2Hz,4H), 3.23(d,J=8.2Hz,2H),3.07(q,J=6.4Hz,4H),2.84(dt,J=6.0,5.4Hz,4H),2.22(s,2H),1.52(t,J=6.6Hz,4H),1.07(d,J=6.6Hz,6H).

[0643] 18. Synthesis of D-30 compound

[0644]

[0645] D-5 (23.4 mg, 37.6 μmol) prepared in part 3 of this example was dissolved in 12 mL of DMF, HATU (14.3 mg, 37.6 μmol), 3-amino-1-propanol (2.8 mg, 37.6 μmol), and diisopropylamine (4.8 mg, 37.6 μmol) were added to the solution, and the mixture was reacted at 25° C. for 4 hours. After the reaction solution was concentrated, the crude product was purified by flash column chromatography to obtain white solid D-30 (4.8 mg, 7.1 μmol, 19% yield).

[0646] D-30:MS m / z(ESI):680.6[M+H] + ; 1 H NMR (600MHz, DMSO-d6) δ12.11 (s, 1H), 7.83 (t, J = 5.7Hz,

[0647] 1H),7.35(s,1H),7.32(s,1H),7.18(d,J=1.7Hz,3H),7.05(d,J=5.6Hz,2H),4.16(s,4H ),3.95(d,J=5.2Hz,6H),3.87(d,J=1.5Hz,6H),3.38(d,J=6.3Hz,2H),3.23(dd,J=15.2, 6.9Hz,2H),3.07(td,J=7.0,2.3Hz,2H),2.90–2.79(m,2H),2.23(dd,J=12.3,6.1Hz,2H) ,1.99–1.95(m,2H),1.52(p,J=6.6Hz,2H),1.16(d,J=7.2Hz,3H),1.07(d,J=6.7Hz,3H).

[0648] 19. Synthesis of D-31 compound

[0649]

[0650] D-5 (40 mg, 64.3 umol) prepared in part 3 of this example, N, N'-dicyclohexylcarbodiimide (39.8 mg, 192.9 umol), 4-dimethylaminopyridine (1.6 mg, 12.8 umol), benzyl alcohol (27.8 mg, 257.2 umol) were dissolved in dichloromethane (1 mL). After stirring the reaction solution for 12 hours, the pH was adjusted to about 3 with dilute hydrochloric acid, and then crudely purified with a silica gel column, and further purified with the pre-preparation to obtain D-31 (42 mg, 52.3 umol, 81% yield).

[0651] D-31: 1 H NMR (400MHz, CDCl3) δ7.34–7.27(m,10H),7.17(s,2H),7.11(s,2H),6.71(s,2H),5.13(q,J=12.4Hz,4H),4.30(t,J=6.2Hz,4H),3.99(s,6H ),3.94(s,6H),3.42(dd,J=16.4,7.8Hz,2H),3.27–3.12(m,2H),3.01(dd,J=16.5,5.9Hz,2H),2.43(p,J=6.2Hz,2H),1.29(d,J=7.1Hz,6H); 13C NMR (101MHz, CDCl3) δ190.36,175.85,151.36,145.56,136.11,133.51,128.47,128.06,1 28.02,118.73,111.63,105.13,92.12,66.43,66.36,56.13,42.65,35.54,32.40,17.22.

[0652] 20. Synthesis of D-32 compound

[0653]

[0654] D-5 (40 mg, 64.3 umol) prepared in part 3 of this example, N, N'-dicyclohexylcarbodiimide (39.8 mg, 192.9 umol), 4-dimethylaminopyridine (1.6 mg, 12.8 umol), isopropanol (15.5 mg, 257.2 umol) were dissolved in dichloromethane (1 mL). After stirring the reaction solution for 12 hours, the pH was adjusted to about 3 with dilute hydrochloric acid, and then crudely purified with a silica gel column, and further purified with the pre-prepared solution to obtain D-32 (14 mg, 19.3 umol, 30% yield).

[0655] D-32: 1 H NMR (600MHz, CDCl3) δ7.19 (s, 2H), 7.12 (s, 2H), 6.72 (s, 2H), 5.01 (hept, J = 6.2Hz, 2H),

[0656] 4.30(t,J=6.2Hz,4H),4.02(s,6H),3.94(s,6H),3.38(dd,J=16.4,7.7Hz,2H),3.10–3.02(m,2H), 2.97(dd,J=16.4,6.1Hz,2H),2.43(p,J=6.1Hz,2H),1.26(d,J=7.1Hz,6H),1.22(t,J=6.6Hz,12H); 13 CNMR (151MHz, CDCl3) δ190.60,175.54,151.30,145.54,136.07,133.60,118.73,111.56 ,105.15,92.14,67.69,66.43,56.12,42.68,35.70,32.40,29.31,21.76,21.74,17.24.

[0657] 21. Synthesis of D-33 compound

[0658]

[0659] D-5 (40 mg, 64.3 umol) prepared in part 3 of this example, N, N'-dicyclohexylcarbodiimide (39.8 mg, 192.9 umol), 4-dimethylaminopyridine (1.6 mg, 12.8 umol), isobutyl alcohol (19.0 mg, 257.2 umol) were dissolved in dichloromethane (1 mL). After stirring the reaction solution for 12 hours, the pH was adjusted to about 3 with dilute hydrochloric acid, and then crudely purified with a silica gel column, and further purified with the pre-preparation to obtain D-33 (12 mg, 16.1 umol, 25% yield).

[0660] D-33: 1 H NMR (600MHz, CDCl3) δ7.19 (s, 2H), 7.12 (s, 2H), 6.72 (s, 2H), 4.30 (hept, J = 6.2Hz, 4H),

[0661] 4.02(s,6H),3.94(s,6H),3.88(t,J=6.2Hz,4H),3.41(dd,J=16.4,7.7Hz,2H),3.17–3.10(m,2H),2.99(dd ,J=16.4,6.1Hz,2H),2.48–2.40(m,2H),1.98–1.88(m,2H),1.28(d,J=7.1Hz,6H),0.91(t,J=6.6Hz,12H); 1 H NMR (600MHz, CDCl3) δ7.19,7.12,6.72,4.30,4.30,4.30,4.28,4.01,3.94,3.91,3.90,3.90,3.89,3.88,3.87,3.87,3.86,3.43,3.42,3.4 0,3.39,3.15,3.15,3.14,3.13,3.11,3.10,3.01,3.00,2.98,2.97,2 .44,2.43,2.41,1.95,1.94,1.92,1.90,1.29,1.28,0.91,0.91,0.91.

[0662] 22. Synthesis of positive control compound D-34 and positive control compound D-35

[0663]

[0664] Step 1: To a solution of 2-fluoro-4,5-dimethoxybenzaldehyde (5.0 g, 27.15 mmol) in DMF (100.0 mL), ethyl mercaptoacetate (3.5 g, 32.97 mmol) and K2CO3 (13.4 g, 97.10 mmol) were added and stirred at 60° C. for 16 hours. The mixture was extracted with EtOAc (100.0 mL x 3) and H2O (200.0 mL), the organic phase was dried and concentrated, and purified by column chromatography (petroleum ether: ethyl acetate = 5:1) to give a yellow solid D-34-int-1 (5.6 g, 22.20 mmol, yield 81.8%).

[0665] D-34-int-1:MS m / z(ESI):253.1[M+H]+.

[0666] Step 2: D-34-int-1 (5.6 g, 22.20 mmol) was dissolved in THF (50.0 mL), and a solution of LiOH (4.7 g, 196.24 mmol) in H2O (25.0 mL) was added dropwise at 0 ° C. The reactant was warmed to room temperature from 0 ° C and stirred for 16 h. The reaction was quenched with ice water (100.0 mL), the pH was adjusted to 1-2 with HCl (1 N) at 0 ° C, the mixture was extracted with EtOAc (50.0 mL x 3), and the organic layer was dried and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 1: 1) to give D-34-int-2 (3.2 g, 13.43 mmol, 69.5% yield) as a white solid.

[0667] D-34-int-2: 1 H NMR (400MHz, DMSO-d6) δ7.94(s,1H),7.58(s,1H),7.48(s,1H),3.85(s,3H),3.82(s,3H).

[0668] Step 3: Cu2O (7.7 g, 53.72 mmol) was added to a solution of D-34-int-2 (3.2 g, 13.43 mmol) in DMF (60.0 mL), and the reactant was stirred at 140 ° C for 16 hours. After cooling to room temperature, the mixture was extracted with EtOAc (100.0 mL x 3) and H2O (200.0 mL), and the EtOAc layer was dried and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 30: 1) to give D-34-int-3 (2.4 g, 12.36 mmol, 92.0% yield) as a white solid.

[0669] D-34-int-3:MS m / z(ESI):195.2[M+H] + .

[0670] Step 4: At 0 ° C, 3,3-dimethyltetrahydrofuran-2,5-dione (1.7 g, 13.38 mmol) was added dropwise to a DCM (25.0 mL) solution containing D-34-int-3 (1.3 g, 6.69 mmol) and aluminum chloride (1.8 g, 13.38 mmol). The reactant was stirred from 0 ° C to room temperature for 16 h. The reaction was quenched with ice water (50.0 mL) and extracted with DCM (20 mL x 3). The DCM layer was dried and concentrated to give a light yellow solid D-34-int-4 (1.2 g, 3.72 mmol, 55.6% yield).

[0671] D-34-int-4:MS m / z(ESI):323.1[M+H] + .

[0672] Step 5: To D-34-int-4 (0.8 g, 2.54 mmol), potassium carbonate (703.1 mg, 5.09 mmol, 2.00 eq) in DMF (2.0 mL) was added iodomethane (1.1 g, 7.63 mmol). The reactant was stirred at room temperature for 16 hours. The mixture was diluted in H2O (40.0 mL), extracted with EtOAc (20.0 mL x 3), and the EtOAc layer was dried and concentrated. The residue was purified by column chromatography (petroleum ether: ethyl acetate = 5: 1) to give a yellow solid D-34-int-5 (560.0 mg, 1.66 mmol, 65.5% yield).

[0673] D-34-int-5:MS m / z(ESI):337.1[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.18(s,1H),7.59(s,1H),7.45(s,1H),3.86(s,3H),3.84(s,3H),3.57(s,3H),3.36(s,2H),1.24(s,6H).

[0674] Step 6: Aluminum chloride (4.0 g, 29.73 mmol) was added in batches to a solution of D-34-int-5 (1.0 g, 2.97 mmol) in DCM (20.0 mL) at room temperature, and the reaction was stirred at room temperature for 16 hours. The mixture was diluted in H2O (40.0 mL), extracted with DCM (20.0 mL x 3), and the DCM layer was dried and concentrated. The residue was purified by SFC to obtain D-34-int-6 (158.0 mg, 490.11 umol, 16.5% yield) as a white solid.

[0675] D-34-int-6:MS m / z(ESI):323.1[M+H]+; 1 H NMR (400MHz, DMSO-d6) δ9.40(s,1H),8.14(s,1H),7.54(s,1H),7.31(s,1H),3.87(s,3H),3.57(s,3H),3.34(s,2H),1.24(s,6H).

[0676] Step 7: 1,3-dibromopropane (93.9 mg, 465.30 umol) and K2CO3 (42.9 mg, 310.20 umol) were added to a solution of D-34-int-6 (50.0 mg, 155.10 umol) in MeCN (2.0 mL) and stirred for 16 hours at 60 ° C. After cooling to room temperature, the mixture was concentrated. The residue was purified by preparative TLC (petroleum ether: ethyl acetate = 3: 1) to give D-34-int-7 (67.0 mg, 151.12 umol, 97.4% yield) as a white solid.

[0677] D-34-int-7:MS m / z(ESI):445.1[M+H]+.

[0678] Step 8: D-34-int-7 (57.0 mg, 128.57 umol) was added to a solution of D-34-int-6 (41.5 mg, 128.5 7 umol) and K2CO3 (35.5 mg, 257.13 umol) in MeCN (2.0 mL), and stirred at 60 ° C for 16 hours. After cooling to room temperature, the mixture was concentrated. The residue was purified by preparative TLC (petroleum ether: ethyl acetate = 3: 1) to give a white solid D-34 (60.0 mg, 87.61 umol, 68.2% yield).

[0679] D-34:MS m / z(ESI):685.2[M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ8.14(s,2H),7.61(s,2H),7.51(s,2H),4.23(t,J=6.0H z,4H),3.86(s,6H),3.57(s,6H),3.35(s,4H),2.36-2.21(m,2H),1.24(s,12H).

[0680] Step 9: D-34 (55.0 mg, 80.31 umol) is dissolved in THF (2.0 mL), MeOH (1.0 mL), and LiOH (9.6 mg, 401.57 umol) is added dropwise in H2O (1.0 mL) solution. The mixture is stirred at room temperature for 16 h. The concentrated mixture is diluted with ice water (5.0 mL), pH is adjusted to 1-2 with HCl (1 N), the mixture is extracted with EtOAc (5.0 mL x 3), and the EtOAc layer is dried and concentrated. The residue is purified by preparative HPLC to obtain D-35 (1.8 mg, 2.80 umol, 3.5% yield) as a white solid.

[0681] D-35:MS m / z(ESI):655.0[MH] - ; 1 H NMR(400MHz,DMSO-d6)δ12.05(s,2H),8.13(s,2H),7.60(s,2H),7.52(s,2H), 4.23(s,4H),3.86(s,6H),3.29(s,4H),2.31(d,J=15.8Hz,2H),1.23(s,12H).

[0682] 23. Synthesis of D-36 and D-37 compounds

[0683]

[0684] Step 1: Dissolve intermediate M-29 (3.05 g, 10 mmol), potassium carbonate (3.45 g, 25 mmol), and benzyl bromide (1.43 mL, 12 mmol) in acetonitrile (20 mL). Stir the reaction solution for 12 hours and purify it with a silica gel column to obtain D-36-int-1 (3.6 g, 9.11 mmol, yield 91%).

[0685] Step 2: D-36-int-1 (3.6 g, 9.11 mmol) and lithium hydroxide (875 mg, 36.4 mmol) were dissolved in THF / MeOH / H2O (50 mL / 25 mL / 25 mL) and stirred for 12 hours. After the reaction solution was concentrated, it was directly purified by flash column chromatography (300-400 mesh silica gel, dichloromethane / methanol = 20 / 1) to obtain D-36-int-1 (3.2 g, 8.4 mmol, yield 92%).

[0686] Step 3: D-36-int-1 (200 mg, 525 umol), 1-hydroxybenzotriazole (85.3 mg, 631 umol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (118.1 mg, 616 umol) were dissolved in dichloromethane (1.6 mL) and activated for 10 minutes. Then ammonium chloride (41.7 mg, 780 umol) and triethylamine (108 uL, 777 umol) were added to the above reaction solution. After stirring the reaction solution for 12 hours, D-36-int-2 (131 mg, 345 umol, yield 66%) was obtained by crude purification with a silica gel column.

[0687] Step 4: Add a mixed solution of D-36-int-2 (131 mg, 345 umol) and methanol / dichloromethane (5.0 mL / 0.5 mL) to a vial, add a drop of water under stirring, and then add palladium / carbon (13.1 mg, 10% wt in 55% H2O). Insert a hydrogen balloon into the reaction apparatus, and stir the system at 23°C for 2.0-2.5 hours. The reaction mixture is directly filtered with a sand core funnel containing diatomaceous earth, rinsed with dichloromethane, and the filtrate is concentrated and directly subjected to rapid column chromatography (300-400 mesh silica gel, methanol / dichloromethane = 10 / 90) to obtain D-36-int-3 (78.3 mg, 270 umol, yield 78%).

[0688] Step 5: D-36-int-3 (78.3 mg, 270 umol), anhydrous N,N'-dimethylformamide (0.5 mL) and anhydrous potassium carbonate (111.8 mg, 810 umol) were added to a vial. The reaction system was stirred at 65 ° C for 1.0 hour, and then an anhydrous N, N'-dimethylformamide solution (0.5 mL) of D-1-int-1 (172 mg, 405 umol) obtained in step 1 was added thereto, and stirring was continued at 65 ° C for 16 hours, diluted with 10 mL of ethyl acetate, quenched by adding 10 mL of water dropwise, and then adjusted to pH = 4-5 with 1.0 M dilute hydrochloric acid, extracted with ethyl acetate (30 mL×3), combined organic phases, washed with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, concentrated, and directly purified by flash column chromatography (300-400 mesh silica gel, methanol / dichloromethane = 5 / 95) to obtain D-36 (78.3 mg, 123 umol, 46% yield).

[0689] D-36: 1 H NMR (400MHz, CDCl3) δ7.20 (s, 1H), 7.18 (s, 1H), 7.11 (s, 2H), 6.71 (d, J = 3.5Hz, 2H), 5.86

[0690] (brs,1H),5.31(brs,1H),4.29(t,J=6.0Hz,4H),4.02(s,6H),3.94(s,6H),3.71(s,3H),3.49–3.35(m,2H),3.11(dq, J=14.4,7.2Hz,1H),3.02(qd,J=12.5,4.9Hz,3H),2.43(p,J=6.2Hz,2H),1.28(d,J=1.4Hz,3H),1.27(d,J=1.9Hz,3H); 13 C NMR (101MHz, CDCl3) δ191.11,190.43,178.35,176.61,151.45,151.35,145.59,145.55,136.21,136.11,133.43,133.38,118.7 5,118.72,112.09,111.62,105.08,92.12,92.05,66.37,56.11,51.91,43.27,42.70,35.75,35.29,32.41,29.28,17.93,17.25.

[0691] Step 6: D-36 (20 mg, 31.5 umol) and lithium hydroxide (3 mg, 125 umol) were dissolved in THF / MeOH / H2O (0.5 mL / 0.25 mL / 0.25 mL) and stirred for 12 hours. After the reaction solution was concentrated, it was directly subjected to rapid column chromatography (300-400 mesh silica gel, dichloromethane / methanol = 20 / 1) to obtain D-37.

[0692] 24. Synthesis of D-38 and D-39 compounds

[0693]

[0694] Step 1: Dissolve D-36-int-1 (100 mg, 253 umol), pyridine (28.8 uL, 356 umol), and hydroxylamine hydrochloride (46.2 mg, 655 umol) obtained in step 23 in ethanol (1.14 mL). Stir the reaction solution at 60 ° C for 12 hours and then purify it with a silica gel column to obtain D-38-int-1 (70 mg, 171 umol, 67% yield).

[0695] Step 2: D-38-int-1 (70 mg, 171 umol), N,N'-dicyclohexylcarbodiimide (52.8 mg, 256 umol), 4-dimethylaminopyridine (2 mg, 16 umol), and acetic acid (14.6 uL, 255 umol) were dissolved in dichloromethane (2 mL). The reaction solution was stirred for 12 hours and crudely purified by silica gel column to obtain D-38-int-2 (64 mg, 142 umol, yield 68%).

[0696] Step 3: Add a mixed solution of D-38-int-2 (64 mg, 142 umol) and methanol / dichloromethane (5.0 mL / 0.5 mL) to a vial, add a drop of water under stirring, and then add palladium / carbon (6.4 mg, 10% wt in 55% H2O). Insert a hydrogen balloon into the reaction device, stir the system at 23°C for 2.0-2.5 hours, and directly filter the reaction mixture with a sand core funnel containing diatomaceous earth, rinse with dichloromethane, concentrate the filtrate, and directly perform rapid column chromatography (300-400 mesh silica gel, petroleum ether / acetone = 4 / 1-3 / 1) to obtain D-38-int-3 (16.9 mg, 46.7 umol, 33% yield).

[0697] Step 4: Add D-38-int-3 (16.9 mg, 46.7 umol), anhydrous N,N'-dimethylformamide (0.5 mL) and anhydrous potassium carbonate (19.3 mg, 140 umol) to a vial. The reaction system was stirred at 65 ° C for 1.0 hour, and then an anhydrous N, N'-dimethylformamide solution (0.5 mL) of D-1-int-1 (30 mg, 70.6 umol) obtained in step 1 was added thereto, and stirring was continued at 65 ° C for 16 hours, diluted with 10 mL of ethyl acetate, quenched by adding 10 mL of water dropwise, and then adjusted to pH = 4-5 with 1.0 M dilute hydrochloric acid, extracted with ethyl acetate (30 mL×3), combined organic phases, washed with water and saturated brine in turn, dried over anhydrous sodium sulfate, filtered, concentrated, and directly purified by flash column chromatography (300-400 mesh silica gel, petroleum ether / acetone = 3 / 1-2 / 1-1 / 1) to obtain D-38 (5.9 mg, 8.3 umol, yield 18%).

[0698] D-38: 1 H NMR (400MHz, CDCl3) δ7.07 (s, 1H), 6.99 (d, J = 2.0Hz, 2H), 6.67 (s, 1H), 6.64 (s, 1H), 6.59

[0699] (s,1H),4.17(t,J=6.2Hz,4H),3.89(s,6H),3.82(s,6H),3.58(s,3H),3.48(s,3H),3.28(dd,J=16.4,7.7Hz,1H),3. 09–2.85(m,4H),2.83–2.72(m,1H),2.29(p,J=6.2Hz,2H),2.14(s,3H),1.27(d,J=7.1Hz,3H),1.22(d,J=7.0Hz,3H).

[0700] Step 5: D-38 (5.4 mg, 7.6 umol) and lithium hydroxide (1.8 mg, 76 umol) were dissolved in THF / MeOH / H2O (0.5 mL / 0.25 mL / 0.25 mL) and stirred for 12 hours. After the reaction solution was concentrated, it was directly purified by flash column chromatography (300-400 mesh silica gel, dichloromethane / methanol = 20 / 1) or by preparative liquid chromatography to obtain D-39 (1.8 mg, 2.8 umol, yield 37%).

[0701] D-39: 1 H NMR (400MHz, CD3CN) δ7.17 (s, 1H), 7.05 (s, 1H), 7.02 (s, 1H), 6.82 (d, J = 1.5Hz, 2H),

[0702] 6.52(s,1H),4.11(q,J=6.4Hz,4H),3.89(s,3H),3.81(s,3H),3.78(s,3H),3.76(s,3H),3.25(dd,J=16.8,8.3Hz,1H),2. 96(t,J=4.2Hz,1H),2.89(ddd,J=21.3,11.8,7.6Hz,4H),2.83–2.72(m,2H),1.14(d,J=7.1Hz,3H),1.03(d,J=6.9Hz,3H).

[0703] Test Example 1: THP-1 cell screening experiment of compounds activating interferon gene stimulator protein (STING)

[0704] THP1-Dual cells are reporter cell lines transformed from human THP-1 cells, and their IRF_Luc reporter system induces secretion of luciferase after activation of the interferon regulatory factor (IRF) pathway. By providing the corresponding substrate and enzyme reaction, the generated fluorescence (Luminescence) signal is detected, and the secretion of luciferase can be determined, further reflecting the intensity of the activation of the IRF pathway. Therefore, when the compound activates the interferon gene stimulating protein (STING) of the cell, its downstream IRF3 pathway is activated, inducing cell luciferase secretion, and the activity of the compound activating STING can be analyzed by detecting the fluorescence intensity. Based on this, the activation effect of the compound prepared in Example 1 on STING protein was detected by THP1-Dual cells, and the specific test method is as follows:

[0705] 1. Use DMSO to dilute the monomer compound prepared in Example 1 or the dimer compound prepared in Example 2, and then use the echo acoustic pipetting system of Labcyte to add a certain amount (10-400 nL) of the compound to each well of the 384-well cell culture plate, so that the highest final concentration of the compound in the culture system (20 μL or 40 μL) is 100 μM or 50 μM, and dilute three times. Ten concentrations are set for each compound, and 3 replicate wells are set for each concentration. The positive control compounds are MSA-2 (purchased from MedChemExpress), diABZI (purchased from MCE and selleckchem, with the specific structural formula as shown below Figure 1 The same volume of DMSO was added to the control group without drug addition (DMSO group). The concentration range of highly active compounds was adjusted according to the situation to calculate EC 50 , three parallel groups in each group.

[0706] 2. Centrifuge at 400 × g for 5 minutes to collect THP1-Dual cells, resuspend in fresh medium, and adjust the concentration to 5 × 10 5 cells / mL. 20μL or 40μL of cells were added to each well, and a blank control group was set up, in which only the same volume of medium without cells was added (Blank group). After incubation in the incubator for 20 hours, the supernatant was centrifuged and the luminescence fluorescence signal was detected.

[0707] 3. After centrifugation at 400 × g for 5 minutes, take 5 μL of supernatant from each well into a new 384-well plate and add 7 μL of substrate Quanti-Luc TMGold (purchased from Invivogen). After incubation at room temperature for 1.5 to 2 hours, the fluorescence signal of each well was detected by a multifunctional fluorescence microplate reader (Biotek Synergy Neo2).

[0708] 4. Calculate the signal increase factor of each compound at each concentration. Fluorescence signal increase factor = (signal value of compound test group - signal value of blank control group) / (signal value of DMSO group - signal value of blank control group). Use GraphPadPrism 8 software to perform four-parameter nonlinear fitting (log (agonist) vs. response) on the log value of concentration and the corresponding fluorescence signal increase factor to generate the half effective concentration EC 50 For specific abbreviation results, please refer to Table 1 (monomer compound prepared in Example 1) and Table 2 (dimer compound prepared in Example 2).

[0709] Among them, the structural formula of MSA-2 in this test example is

[0710] Table 1: EC values ​​of each compound 50 result

[0711] Compound Name <![CDATA[EC 50 (μM)]]> Compound Name <![CDATA[EC 50 ]]> MSA-2 (purchased from MedChemExpress) D M-40 C M-2 C M-41 C M-3 C M-42 C M-10 C M-43 C M-13 C M-44 C M-16 B M-45 C M-17 B M-54 C M-18 C M-66 C M-19 C M-68 B M-24 C M-70 B M-32 C M-71 C M-33 C M-72 B M-34 C M-79 C M-39 C

[0712] A stands for EC 50 <0.1 μM, B indicates EC 50 0.1-0.85μM (excluding 0.85μM), C represents EC 50 0.85-9.9μM (excluding 9.9μM), D represents EC 50 ≥9.9μM.

[0713] Table 2: EC values ​​of each compound 50 result

[0714] Compound Name <![CDATA[EC 50 ]]> Compound Name <![CDATA[EC 50 ]]> diABZI (positive control compound) B D-15 A+ D-34 (positive control compound) A D-16 A+ D-35 (positive control compound) A D-17 A+ D-4 A+ D-18 A+ D-5 A+ D-21 A+ D-7 A+ D-23 A+ D-8 A+ D-25 A+ D-9 A+ D-28 A+ D-10 A+

[0715] A+ means EC 50 <0.01μM (10nM), A represents EC 50 0.01-0.1μM (excluding 0.1μM, 10-100nM), B represents EC 50 0.1-0.85 μM (excluding 0.85 μM).

[0716] Test Example 2: Animal in vivo experiments of compounds

[0717] 1. Anti-tumor effect test of compounds in animals

[0718] 7-week-old female C57BL / 6 mice were subcutaneously inoculated with 1×10 6MC38 cells, wait until the tumor is formed and reaches about 50 mm 3 After the size was measured, the mice were randomly divided into groups of 8. Compound D-18 (prepared in Part 8 of Example 2) was administered via the tail vein and oral administration. The doses of the tail vein administration were 0.01 mg / kg, 0.04 mg / kg and 0.07 mg / kg. Vehicle was a solvent control group (5% DMSO + 5% solutol (polyethylene glycol-15 hydroxystearate) + 90% saline (normal saline), 200 μL / kg). The mice were administered 3 times on the 1st, 4th and 8th days. The mice were observed for about 4 weeks and the size of the tumors was recorded. The results are shown in Figure 2 . 0.5 hours after the last administration, blood was collected from the submandibular vein. Each blood sample collected from the submandibular vein was placed in a tube containing sodium heparin anticoagulant, and then centrifuged at 5000 rpm for 10 minutes at 4°C to separate the plasma. The obtained plasma samples were transferred to centrifuge tubes and stored at -80°C. The blood drug concentration was tested. The results are shown in Figure 3 4 hours after the last administration, blood was collected from the submandibular vein to detect the cytokine IFN-β in the serum sample. The results were as follows Figure 4 The results showed that the compound had a dose-dependent effect and could significantly inhibit tumor growth.

[0719] 2. Cytotoxicity or animal toxicity test of compounds

[0720] Eight 7-week-old female Balb / c mice were administered compound D-18 (prepared in Part 8 of Example 2) via the tail vein. The dosages were 0.1 mg / kg, 0.05 mg / kg, and 0.02 mg / kg, respectively. The mice were administered three times on the 1st, 4th, and 8th days. The mice were observed for 2 weeks to detect changes in body weight and the state of the mice. Blood was collected through the submandibular vein at 2, 4, 8, and 24 hours after the first administration. Each blood sample collected through the submandibular vein was placed in a test tube containing sodium heparin anticoagulant, and then centrifuged at 5000 rpm for 10 minutes at 4°C to separate the plasma. The obtained plasma samples were transferred to centrifuge tubes and stored at -80°C. The weight changes of mice after administration are shown in the figure. Figure 5 The results show that the mice did not significantly lose weight after administration, indicating that compound D-18 had no significant toxicity at the given dose. The results of detecting cytokine IFN-β in serum samples are shown in Figure 6 As shown, the results showed that compound D-18 could activate immune response and promote the release of IFN-β, among which the high dose of 0.1 mg / kg had the most obvious effect.

[0721] 3. Pharmacokinetic data of compounds

[0722] Eight 7-week-old male CD1 mice were injected with a single dose of compound D-18 at 0.07 mg / kg and 0.15 mg / kg via the tail vein. Blood was collected from the mice administered the tail vein through the submandibular vein at 5 minutes, 0.25, 0.5, 1, 2, 4, 8, and 24 hours after administration. Each blood sample collected through the submandibular vein was placed in a test tube containing sodium heparin anticoagulant and then centrifuged at 5000 rpm for 10 minutes at 4°C to separate the plasma. The obtained plasma samples were transferred to centrifuge tubes and stored at -80°C for LC-MS / MS analysis. The average drug concentration-time curve of compound D-18 in mouse blood is shown in Figure 2. Figure 7 shown.

[0723] 4. Compound accumulation in tumors

[0724] 7-week-old female C57BL / 6 mice were subcutaneously inoculated with 1×10 6 MC38 cells, wait until the tumor is formed and reaches about 500mm 3 After size measurement, the mice were randomly divided into groups of 3 each. A single injection of compound D-18 0.1 mg / kg was given through the tail vein. The mice were killed 0.5, 4, 7, and 24 hours after administration, and the tumors were removed. After the tumor tissue was lysed with a lysis buffer, it was centrifuged at 10,000 rpm for 10 minutes at 4°C, and the supernatant was extracted. The obtained supernatant sample was transferred to a centrifuge tube for LC-MS / MS analysis. The average drug concentration-time curve of compound D-18 in mouse tumors is shown in Figure 2. Figure 8 shown.

[0725] Test Example 3: Detection of the binding ability of compounds to STING protein

[0726] The cisbio brand commercial STING binding kit was used to detect the binding activity of the compound and human STING wild-type (WT) protein (Revvity, Catalog No. 64BDSTGPEG), human STING AQ mutant protein (Revvity, Catalog No. 64BDSTGQPEG), human STING H232 mutant protein (Revvity, Catalog No. 64BDSTGHPEG) and mouse STING protein (Cat. No. 64BDSTGMPEG). The principle of this kit is fluorescence resonance energy transfer (FRET). The kit mainly contains three components, namely, STING protein with 6 histone (6his) tags, terbium-labeled specific anti-6His antibody as energy donor and d2-labeled STING ligand as energy acceptor. When the donor and the acceptor are close to each other, energy is transferred from the donor to the acceptor, causing the latter to emit fluorescence, and the microplate reader reads and detects a high signal value. When the compound is added, the STING ligand labeled with d2 is replaced, energy transfer cannot occur, the fluorescence intensity is weak, and the detection signal value is low.

[0727] Carry out the following specific experimental operations according to the product instructions:

[0728] 1. Dilute 5× Dilution Buffer and 5× Detection Buffer to 1× using ultrapure water.

[0729] 2. Add 2.5 μL of 1× Dilution Buffer to each well of a white 384-well shallow plate.

[0730] 3. On a 384PP plate or 384LDV plate, use DMSO to dilute the monomer compound prepared in Example 1 or the dimer compound prepared in Example 2 by 2 to 5 times gradient, 6-12 different concentration points. According to the final concentration required by the experiment, use the echo acoustic pipetting system of Labcyte to add a certain amount (2.5-250nL) of compound to each well of the white 384 shallow well plate, so that the DMSO content in the 10μL reaction system does not exceed 2.5%. The positive control compound is diABZI (purchased from MCE and selleckchem, with the specific structural formula as shown below) Figure 1 shown).

[0731] 4. Dilute 50× 6His STING protein to 1× using 1× detection buffer and add 2.5 μL 6HisSTING protein to each well.

[0732] 5. Use 1× detection buffer to dilute 50×6His antibody to 1×, and use 1× detection buffer to dilute d2-labeled STING ligand to 1×. After mixing the two in a 1:1 ratio, add 5 μL of the mixture to each well.

[0733] 6. After sealing the plate with plastic film, incubate at room temperature for 3 hours or overnight. Detect the fluorescence signals at 665nm and 620nm with a multifunctional microplate reader. Calculate the ratio = (665nm signal value / 620nm signal value) × 10 4 , X-axis is the log value of concentration, Y-axis is the ratio, GraphPad Prism8 fitting IC 50 The test results are shown in Table 3.

[0734] Table 3: Competitive binding IC of each compound to different STING proteins 50 result

[0735]

[0736]

[0737] A+ indicates IC 50 <0.01μM (10nM), A represents IC 500.01-0.1μM (excluding 0.1μM, 10-100nM), B represents IC 50 0.1-0.85μM (excluding 0.85μM), C represents IC 50 0.85-9.9μM (excluding 9.9μM), D represents IC 50 ≥9.9μM, N / A means not tested.

[0738] Test Example 4: Tumor target cell and PBMC co-culture system to detect the killing ability of compounds on tumor cells

[0739] STING agonists have strong anti-tumor activity by activating the immune system. In in vitro experiments, by co-culturing with human peripheral blood mononuclear cells (PBMC), it can be detected that STING agonists activate PBMC to kill tumor cells. The specific experimental steps are as follows:

[0740] 1. One day in advance, cryopreserved healthy human PBMC cells (purchased from Shanghai Aoneng Biotechnology Co., Ltd.) were revived and cultured overnight in a RPMI-1640 complete medium, 5% CO2, 37°C cell culture incubator.

[0741] 2. On a 384PP plate or a 384LDV plate, use DMSO to dilute the monomer compound prepared in Example 1 or the dimer compound prepared in Example 2 in a 5-fold gradient. Use the echo acoustic liquid transfer system of Labcyte to add 40 nL of compound to each well of a white 384 shallow well plate at 9 different concentration points (40 μL system final concentration 1000 nM to 0.00256 nM); add 40 nL DMSO to the experimental control group. The positive control compound is diABZI (purchased from MCE and selleckchem, with the specific structural formula as shown below) Figure 1 shown).

[0742] 3. In addition to the compound, 20 μL of 2000 tumor cells were plated in each well of the 384-well plate, and 20 μL of five times the amount of PBMC (10,000 cells / well) was added, and the cells were incubated in a cell culture incubator for 48 hours. At the same time, the "experimental group DMSO control" was set up: containing 40nLDMSO, 20 μLPBMC, 20 μL tumor cells (such as bladder cancer cell T24, bladder cancer cell RT-4, bladder cancer cell UM-UC-3, bladder cancer cell 5637, breast cancer cell SK-BR-3, breast cancer cell MDA-MB-231, breast cancer cell HCC1954, colon cancer cell HT-29, sarcoma cell HT-1080, ovarian cancer cell SK-OV-3, gastric cancer cell SNU-5, lung cancer cell HCC827, rectal cancer, thyroid cancer cell SW579, brain tumor cell SF126, neuroblastoma cell SH-SY5Y, glioma cell U251, head and neck cancer cell Detroit 562, oral squamous cell carcinoma HSC-2, renal carcinoma cell 769-P, liver cancer HuH-7, bile duct carcinoma cell HCCC-9810, leukemia cell K562, lymphoma cell Raji, myeloma cell MM1.S, prostate cancer cell PC-3, pancreatic cancer cell AsPC-1, melanoma cell A-375, cervical cancer cell Hela, endometrial AN3-CA, skin cancer cell A2058, esophageal cancer cell TE-1, rhabdomyosarcoma cell A-673); set up "PBMC group": containing 40nL compound, 20μL PBMC, 20μL cell culture medium, without tumor cells; set up "PBMC group DMSO control": 40nL DMSO, 20μL PBMC, 20μL cell culture medium, without tumor cells.

[0743] 4. Add 20 μL CTG (purchased from Promega) to each well, and detect the fluorescence signal with a multifunctional microplate reader. The tumor cell survival rate corresponding to the same concentration of compound % = (signal value of experimental group - signal value of PBMC group) / (signal value of DMSO control in experimental group - signal value of DMSO control in PBMC group) × 100%, with the X-axis being the log value of the concentration and the Y-axis being the tumor cell survival rate. GraphPadPrism 8 was used to fit the IC 50 The killing ability of the compounds on tumor cells was evaluated. Some of the results are shown in Table 4.

[0744] Table 4: IC of each compound in activating PBMC to kill tumor cells 50 result

[0745]

[0746]

[0747] A+ indicates IC50 <0.01μM (10nM), A represents IC 50 0.01-0.1μM (excluding 0.1μM, 10-100nM), B represents IC 50 0.1-0.85μM (excluding 0.85μM), C represents IC 50 0.85-9.9μM (excluding 9.9μM), D represents IC 50 ≥9.9μM, N / A means not tested.

[0748] In the description of this specification, the description with reference to the terms "one embodiment", "some embodiments", "example", "specific example", or "some examples" etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representations of the above terms do not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described may be combined in any one or more embodiments or examples in a suitable manner. In addition, those skilled in the art may combine and combine the different embodiments or examples described in this specification and the features of the different embodiments or examples, without contradiction.

[0749] Although the embodiments of the present invention have been shown and described above, it is to be understood that the above embodiments are exemplary and are not to be construed as limitations of the present invention. A person skilled in the art may change, modify, replace and vary the above embodiments within the scope of the present invention.

Claims

1. A compound, which is a compound represented by formula (I) or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the compound represented by formula (I): in, R7 and R8 are each independently selected from H, halogen, -CN, -OR 14 、-NHR 14 or -R 14 ; or, R7 and R8 are linked together with the atoms to form a 4-7 membered heterocyclic ring, wherein the 4-7 membered heterocyclic ring contains at least one heteroatom selected from O, N and S; or one of R7 and R8 is linked to A and connected; R7' and R8' are each independently selected from H, halogen, -CN, -OR 14 、-NHR 14 or -R 14 ; A is empty or -C 1～6 Alkylene-, wherein the -C 1～6 Alkylene - optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NHC(O)-C 1～3 Alkyl, -NO2 or -CN substitution; R1 and R1' are each independently empty, optionally substituted with one or more R9 -C 1～6 Alkylene-, -C(O)-C optionally substituted with one or more R9 1～6 Alkylene-, -C(NOH)-C optionally substituted with one or more R9 2～5 alkylene, -C(O)-C optionally substituted by one or more R9 1～6 Alkyleneoxy-, -C(O)-C optionally substituted by one or more R9 1～6 Cycloalkylene-, -C(O)-C optionally substituted by one or more R9 2～6 Alkenylene-, -C(O)-C optionally substituted with one or more R9 2～6 Alkynylidene- or -C(O)-NR 10 R 11 -; R2 and R2' are each independently -CN, -C(O)-, -C(O)R 12 、-C(O)N(R 12 ) a 2- or 4- to 7-membered heterocyclic ring, wherein the 4- to 7-membered heterocyclic ring contains at least one heteroatom selected from O, N and S; R3, R3', R4 and R4' are each independently H, halogen, optionally substituted with one or more R 13 Replaced-C 1～6 Alkyl, optionally with one or more R 13 Replaced-C 1～6 Alkoxy, optionally with one or more R 13 Replaced-C 2～6 alkenyl, optionally substituted with one or more R 13 Replaced-C 2～6 Alkynyl, optionally substituted with one or more R 13 Substituted -C(O)-C 1～6 Alkyl, optionally with one or more R 13 Substituted -C(O)-C 1～6 Alkoxy, optionally with one or more R 13 Substituted -C(O)-C 2～6 alkenyl, optionally substituted with one or more R 13 Substituted -C(O)-C 2～6 Alkynyl, optionally substituted with one or more R 13 Replaced-C 0～6 Alkyl-C(O)OH, optionally with one or more R 13 Replaced-C 1～6 Alkoxy-C(O)OH, optionally with one or more R 13 Replaced-C 2～6 Alkenyl -C(O)OH or optionally substituted by one or more R 13 Replaced-C 2～6 Alkynyl-C(O)OH; R5, R5', R6 and R6' are each independently H, halogen, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 1～6 Cycloalkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl; Each R9 is independently selected from halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl or -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 10 Independently selected from H, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OR 15 , -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 11 independently selected from H, halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution; Each R 12 Independently selected from H, -OR 15 , -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, -C 5～7 Aryl or 4 to 7 membered ring, wherein the -OR 15 , -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 5～7 Aryl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)-, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution; Each R 13 independently selected from halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, -C 1～6 Alkyl and -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 14 are independently selected from H, -Bn, halogen, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkenyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 15 Independently selected from H or -C 1～3 alkyl; X1 and X2 are each independently C or N, and X1 and X2 are not C at the same time; X3, X4 and X5 are each independently C or N; or one of X1 and X2, X3 and R1 together with the connected atoms are linked to form a 4-7 membered heterocyclic ring, and the 4-7 membered heterocyclic ring contains N or Se.

2. The compound according to claim 1, characterized in that R1 and R1' are each independently -C optionally substituted with one or more R9 1～6 Alkylene-, -C(O)-C optionally substituted with one or more R9 1～6 alkylene, -C(O)-C optionally substituted by one or more R9 1～6 Cycloalkylene or -C(O)-NR 10 R 11 -; or, R1 and R1' are each independently -C(O)-NR 10 R 11 -, one of X1 and X2, X3 and the N of R1 are linked together with the atoms connected thereto to form a 4- to 7-membered heterocyclic ring.

3. The compound according to claim 2, characterized in that Each R9 is independently selected from halogen, -C 1～3 Alkyl and -C 1～3 Alkoxy, wherein the -C 1～3 Alkyl and -C 1～3 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution.

4. The compound according to claim 2, characterized in that Each R9 is independently selected from halogen, -C 1～3 Alkyl or -C 1～3 Alkoxy.

5. The compound according to claim 2, characterized in that Each R 10 Independently selected from -C 1～4 Alkyl, wherein the -C 1～4 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2, -CN or -C 1～2 Alkyl substitution.

6. The compound according to claim 2, characterized in that Each R 11 are independently selected from H, halogen, -C 1～4 Alkyl, wherein the -C 1～4 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

7. The compound according to claim 1, characterized in that R2 is -C(O)R 12 、-C(O)N(R 12 ) a 2- or 4- to 7-membered heterocyclic ring, wherein the 4- to 7-membered heterocyclic ring contains at least one heteroatom selected from O, N and S.

8. The compound according to claim 7, characterized in that Each R 12 Independently selected from H, -OR 15 or -C 1～3 Alkyl, wherein the -OR 15 or -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN or phenyl.

9. The compound according to claim 8, characterized in that Each R 15 Independently selected from H or -C 1～3 alkyl.

10. The compound according to claim 1, characterized in that R3 and R4 are each independently H, halogen, -C optionally substituted with one or more -OH 1～3 Alkyl or -C 0～2 Alkyl-C(O)OH.

11. The compound according to claim 1, characterized in that R5 and R6 are each independently H, halogen or -C 1～3 alkyl.

12. The compound according to claim 1, characterized in that R7 and R8 are each independently -OR 14 ; Or, R7 and R8 are linked together with the atoms to which they are connected to form a 4- to 7-membered heterocyclic ring containing at least one heteroatom selected from O, N and S.

13. The compound according to claim 12, characterized in that Each R 14 are independently selected from H, -C optionally substituted with one or more halogens 1～3 Alkyl or -C optionally substituted by one or more halogen 2～3 Alkenyl.

14. The compound according to claim 1, characterized in that X1 is C, X2 is N or Se, or X1 is N or Se, and X2 is C.

15. The compound according to claim 1, characterized in that X3, X4 and X5 are C.

16. The compound according to claim 1, characterized in that The compound is a compound represented by formula (Ia) or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the compound represented by formula (Ia): Wherein, R1 is -C(OH)-C 1～5 Alkylene, -C(O)-C 2～5 Alkylene, -C(O)-C 2～4 Cycloalkylene, -C(O)-NR 10 R 11 -or-C(NOH)-C 2～5 Alkylene; R 10 is H or -CH3; R 11 For -C 2～4 alkyl; R2 is -C(O)OH, -C(O)OCH3, -C(O)OCH2CH3, -C(O)N(H)OCH3, -C(O)N(H)(CH2)2OH or R3 is H, -CH3, -CH2CH3, -C(O)CH3, -C 1～5 Alkylene -C(O)OH or -C(O)OH; R4, R5 and R6 are each independently H or halogen; R7 and R8 are each independently -OH, -OCH3 optionally substituted by halogen, or -OCH2CH=CH2.

17. The compound according to any one of claims 1 to 16, characterized in that -R1-R2 has the following structure: and -C(O)OH.

18. The compound according to claim 1, characterized in that The compound is a stereoisomer, a tautomer, a solvate, or a pharmaceutically acceptable salt of the compound represented by formula (VI): Among them, R 1a is hydrogen or -CH3; R 1b and R 1c are each independently H or halogen; R 1d is -OH, -OCH3 or -OCH2CH3; R 1f -CH3 or -CH2-CH ＝ CH2; R 1g for Among them, R 1f for When R 1d Not -OH.

19. A compound, which is a compound represented by the following structure or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the compound represented by the following structure:

20. A dimer compound, which is a dimer compound represented by formula (II), formula (III) or formula (IV), or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the dimer compound represented by formula (II), formula (III) or formula (IV): in, A is empty or -C 1～6 Alkylene-, wherein the -C 1～6 Alkylene - optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NHC(O)-C 1～3 Alkyl, -NO2 or -CN substitution; R1 and R1' are each independently empty, optionally substituted with one or more R9 -C(O)-C 1～6 Alkylene-, -C(NOH)-C optionally substituted with one or more R9 2～5 alkylene, -C(O)-C optionally substituted by one or more R9 1～6 Alkyleneoxy-, -C(O)-C optionally substituted by one or more R9 1～6 Cycloalkylene-, -C(O)-C optionally substituted by one or more R9 2～6 Alkenylene-, -C(O)-C optionally substituted with one or more R9 2～6 Alkynylidene- or -C(O)-NR 10 R 11 -; R2 and R2' are each independently -CN, -C(O)-, -C(O)R 12 、-C(O)N(R 12 ) a 2- or 4- to 7-membered heterocyclic ring, wherein the 4- to 7-membered heterocyclic ring contains at least one heteroatom selected from O, N and S; R3, R3', R4 and R4' are each independently H, halogen, optionally substituted with one or more R 13 Replaced-C 1～6 Alkyl, optionally with one or more R 13 Replaced-C 1～6 Alkoxy, optionally with one or more R 13 Replaced-C 2～6 alkenyl, optionally substituted with one or more R 13 Replaced-C 2～6 Alkynyl, optionally substituted with one or more R 13 Substituted -C(O)-C 1～6 Alkyl, optionally with one or more R 13 Substituted -C(O)-C 1～6 Alkoxy, optionally with one or more R 13 Substituted -C(O)-C 2～6 alkenyl, optionally substituted with one or more R 13 Substituted -C(O)-C 2～6 Alkynyl, optionally substituted with one or more R 13 Replaced-C 0～6 Alkyl-C(O)OH, optionally with one or more R 13 Replaced-C 1～6 Alkoxy-C(O)OH, optionally with one or more R 13 Replaced-C 2～6 Alkenyl -C(O)OH or optionally substituted by one or more R 13 Replaced-C 2～6 Alkynyl-C(O)OH; R5, R5', R6 and R6' are each independently H, halogen, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 1～6 Cycloalkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl; R7, R7', R8 and R8' are each independently H, halogen, -CN, -OR 14 、-NHR 14 or -R 14 ; Each R9 is independently selected from H, halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl or -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 10 Independently selected from H, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OR 15 , -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 11 independently selected from H, halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 1～6 Alkoxy, -C 2～6 Alkenyl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution; Each R 12 Independently selected from H, -OR 15 , -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 2～6 Alkynyl, -C 5～7 Aryl or 4 to 7 membered ring, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl, -C 5～7 Aryl or -C 2～6 The alkynyl group is optionally substituted with one or more halogen, -OH, -C(O)-, -C(O)OH, -SH, -NH2, -NO2, -CN, phenyl, -C 1～3 Alkyl and -C 1～3 Alkoxy substitution; Each R 13 independently selected from halogen, -OH, -C(O)OH, -SH, -NH2, -NO2, -CN, -C 1～6 Alkyl and -C 1～6 Alkoxy, wherein the -C 1～6 Alkyl and -C 1～6 The alkoxy group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 14 are independently selected from H, -Bn, halogen, -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 Alkynyl, wherein the -C 1～6 Alkyl, -C 2～6 Alkenyl or -C 2～6 The alkenyl group is optionally substituted with one or more halogen, -OH, -C(O)OH, -C(O)-, -SH, -NH2, -NO2, -CN, -C 1～3 Alkyl or -C 1～3 Alkoxy substitution; Each R 15 Independently selected from H or -C 1～3 alkyl; X1 and X2 are each independently C, N, S or Se, and X1 and X2 are not C at the same time; X3, X4 and X5 are each independently C or N.

21. The dimer compound according to claim 20, characterized in that R1 and R1' are each independently -C(O)-C optionally substituted with one or more R9 1～6 Alkylene-, optionally substituted with one or more R9--C(NOH)-C 1～6 Alkylene or -C(O)-NR 10 R 11 -.

22. The dimer compound according to claim 21, characterized in that Each R9 is independently selected from halogen and -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

23. The dimer compound according to claim 21, characterized in that Each R 10 Independently selected from -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

24. The dimer compound according to claim 21, characterized in that Each R 11 Independently selected from H or -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

25. The dimer compound according to claim 20, characterized in that R2 and R2' are each independently -C(O)R 12 or -C(O)N(R 12 )2.

26. The dimer compound according to claim 25, characterized in that Each R 12 Independently selected from H, -OR 15 or -C 1～3 Alkyl, wherein the -C 1～3 The alkyl group is optionally substituted with one or more halogen, -OH, -SH, -NH2, -NO2 or -CN.

27. The dimer compound according to claim 25, characterized in that Each R 15 Independently selected from H or -C 1～3 alkyl.

28. The dimer compound according to claim 20, characterized in that R3, R3', R4, R4', R5, R5', R6 and R6' are each independently H, halogen or -C 1～3 alkyl.

29. The dimer compound according to claim 20, characterized in that R7, R7', R8 and R8' are each independently -C 1～3 Alkoxy.

30. The dimer compound according to claim 20, characterized in that A is empty or -C 1～4 Alkylene, wherein the -C 1～4 Alkylene - optionally substituted with one or more halogen, -OH, -SH, -NH2, -NHC(O)-C 1～3 Alkyl, -NO2 or -CN substituted.

31. The dimer compound according to claim 20, characterized in that The dimer compound is a dimer compound represented by formula (V), or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt of the dimer compound represented by formula (V): Among them, A is R 2a and R 2b Each independently And R 2a and R 2b Not containing N at the same time; R 2c and R 2d Each is independently -OH, -NH2, -OCH3, -OCH2CH3, -OCH2(CH3)2, -OCH2CH2(CH3)2 or C 5～7 Aryloxy.

32. A dimer compound, which is a dimer compound shown in the following structure or a stereoisomer, a tautomer, a solvate, or a pharmaceutically acceptable salt of the dimer compound shown in the following structure:

33. Use of the compound according to any one of claims 1 to 19 or the dimer compound according to any one of claims 20 to 32 as a STING agonist.

34. A STING agonist, characterized in that The invention comprises the compound according to any one of claims 1 to 19 or the dimer compound according to any one of claims 20 to 32, and optionally a pharmaceutically acceptable carrier, excipient, or vehicle.

35. A pharmaceutical composition, characterized in that It comprises the compound according to any one of claims 1 to 19, the dimer compound according to any one of claims 20 to 32, or the STING agonist according to claim 34, and optionally a pharmaceutically acceptable carrier, excipient, or vehicle.

36. Use of the compound of any one of claims 1 to 19, the dimer compound of any one of claims 20 to 32, the STING agonist of claim 34, or the pharmaceutical composition of claim 35 in the preparation of a medicament for preventing or treating a cell proliferative disease; Optionally, the cell proliferative disorder is a tumor or cancer.

Images