Aryl phosphine oxide compounds and their uses

Abstract

The present invention provides arylphosphine oxides with kinase inhibitory activity, which can effectively inhibit the activities of various types of EGFR resistant mutants (such as EGFR del19 , EGFR del19 / T790M , EGFR del19 / C797S , EGFR T790M / L858R , EGFR L858R / C797S , EGFR del19 / T790M / C797S , EGFR L858R / T790M / C797S ), and also has a significant inhibitory effect on the ALK fusion gene and mutants (such as L1196M), and can be used for the treatment, combination therapy or prevention of various cancers.

Description

[0001] This application is a divisional application of Chinese Patent Application No. 202180004570.6, with an application date of July 2, 2021 and an invention title of "Aryl Phosphoryl Compounds and Their Uses". Technical Field

[0002] The present invention relates to the field of pharmaceuticals. Specifically, it relates to aryl phosphoryl compounds as protein kinase inhibitors. Background Art

[0003] Protein kinases represent a large family of proteins that play important roles in the regulation of various cellular processes and the maintenance and control of cellular functions. They include proliferation, apoptosis, cytoskeletal rearrangement, differentiation, development, immune response, nervous system function and conduction. Additionally, many diseases and / or dysfunctions are associated with the abnormal, aberrant or dysregulated activity of one or more kinases.

[0004] Lung cancer is one of the most common malignancies, generally divided into two major categories: small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC). However, regardless of geographical location, whether in China or globally, lung cancer ranks first. Among them, non-small cell lung cancer (NSCLC) accounts for more than 80% of the total number of lung cancers, seriously threatening human health (Chinese Journal of Lung Cancer [J], Feb 20, 2012; 15(2): 106-111).

[0005] EGFR (epidermal growth factor receptor) is named epidermal growth factor receptor in full. It is a transmembrane glycoprotein with tyrosine kinase activity that is widely distributed on the cell membranes of various tissues in the human body. The mutation and abnormal activation of EGFR are closely related to the occurrence, development, malignancy, and metastasis of various tumors such as non-small cell lung cancer, breast cancer, and esophageal cancer. Most patients with non-small cell lung cancer (NSCLC) have overexpression of EGFR. In the Asian NSCLC population (especially the Chinese population), approximately 40-50% have EGFR mutations. Therefore, the inhibition of EGFR can significantly improve the survival period of NSCLC patients. The common mutations of EGFR can be divided into two categories. One is drug-sensitive mutations, that is, anti-tumor targeted drugs can be used after mutation, such as exon 19 deletion and L858R mutation in exon 21. The other is drug-resistant mutations, that is, resistance to a certain anti-tumor targeted drug after mutation, such as T790M mutation and C797S mutation. The first-generation EGFR small molecule inhibitor drugs Gefitinib, Erlotinib, and Icotinib have achieved significant clinical efficacy in patients carrying EGFR-sensitive mutations and extended the survival period. However, most patients will develop drug resistance after using the drugs for several months. Among them, more than 50% of drug-resistant patients develop drug resistance due to the T790M mutation of EGFR. Although the second-generation EGFR irreversible inhibitor drugs Afatinib and Neratinib have obtained good results in preclinical studies, they lack selectivity for wild-type EGFR (EGFRWT) and have side effects such as large skin toxicity. The third-generation irreversible inhibitor Osimertinib (AZD9291) that overcomes EGFR T790M drug resistance can effectively treat advanced non-small cell lung cancer patients with epidermal growth factor receptor T790M mutation or resistant to other EGFR inhibitors in clinical practice. Although Osimertinib has achieved great success in the clinical treatment of NSCLC with EGFRT790M mutation, some benefited patients have developed drug resistance again after 9-14 months of treatment (Nature Medicine 2015, 21(6), 560-562). It has been found through research that up to 40% of drug-resistant patients develop Osimertinib drug resistance due to the (EGFR)C797S point mutation. Further mechanism studies have shown that the point mutation of (EGFR)C797S converts cysteine at position 797 to serine, resulting in the inability of Osimertinib to form a covalent bond with the target protein, ultimately leading to drug resistance. Currently, there is no drug in clinical practice that can effectively inhibit EGFR for the new mutation (C797S).Therefore, there is an urgent need for new types of highly selective EGFR inhibitors to address issues such as drug resistance caused by the (EGFR)C797S point mutation.

[0006] Anaplastic lymphoma kinase (ALK), also known as ALK tyrosine kinase receptor or CD246, is an active enzyme encoded by the ALK gene in the human body. The fusion genes formed by ALK are closely related to the occurrence and development of various tumors such as non-small cell lung cancer. In patients with non-small cell lung cancer, fusion oncogenes such as EML4-ALK (echinoderm microtubule-associated protein 4 and anaplastic lymphoma kinase fusion gene) account for approximately 3-7%. Therefore, developing protein kinase inhibitors targeting ALK fusion gene-positive has important clinical value. On the other hand, in recent years, with the increase in the number of non-small cell lung cancer patients and the popularization of the application of next-generation sequencing technology (deep sequencing), researchers have continuously found that in some non-small cell lung cancer patients, the phenomenon of simultaneous occurrence of EGFR mutants and ALK gene fusion can occur. Therefore, there is an urgent need for new types of highly selective protein kinase inhibitors to address issues such as drug resistance caused by the (EGFR)C797S point mutation, and at the same time, to address ALK gene fusion and mutations. Summary of the Invention

[0007] One or more embodiments of the present application provide aryl phosphoxy compounds or pharmaceutically acceptable salts thereof shown by the following formula (I) as kinase inhibitors. Such compounds can effectively inhibit the activities of various types of EGFR resistant mutants (such as EGFR del19 、EGFR del19 / T790M 、EGFR del19 / C797S 、EGFR T790M / L858R 、EGFR L858R / C797S 、EGFR del19 / T790M / C797S 、EGFR L858R / T790M / C797S ), and also have significant inhibitory effects on ALK fusion genes and mutants (such as L1196M), and can be used for the treatment, combination treatment, or prevention of various cancers.

[0008]

[0009] Wherein, R1 is selected from C1-C6 alkyl and C3-C6 cycloalkyl, and the C1-C6 alkyl and C3-C6 cycloalkyl are optionally substituted by 0-6 R';

[0010] R2 is selected from hydrogen, amino, C1-C6 alkyl, C3-C6 cycloalkyl, C3-C6 cycloalkenyl, phenyl, 5-6 membered heteroaryl, The amino, C1-C6 alkyl, C3-C6 cycloalkyl, phenyl, 5-6 membered heteroaryl are optionally substituted by 0-3 R aGroup substitution;

[0011] X is selected from CH, S, N or O;

[0012] n = 0, 1 or 2;

[0013] When X is O, Ra is absent;

[0014] ----- bond represents a C-C saturated bond or a C═C alkene bond;

[0015] R3 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, -NR b R c and halogen; the C1-C6 alkyl, C3-C6 cycloalkyl and -NR b R c is optionally substituted by 0-3 R';

[0016] R4 and R6 are each independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl and halogen, and the C1-C6 alkyl and C3-C6 cycloalkyl are optionally substituted by 0-3 R';

[0017] R5 is selected from C1-C6 alkyl and halogen, and the C1-C6 alkyl is optionally substituted by 0-3 R';

[0018] R a R aa are each independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, phenyl, 5-6 membered heteroaryl, halogen, amino, hydroxy, cyano, nitro, -NR b C(O)R c -NR b R c and wherein the C1-C6 alkyl, C3-C6 cycloalkyl, phenyl, 5-6 membered heteroaryl, amino and hydroxy are optionally substituted by 0-3 R';

[0019] Y is selected from N or O, Z is selected from CH or N; when Y is O, R' is absent;

[0020] R b R c are each independently selected from hydrogen, C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, hydroxy and amino; the C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl and C2-C6 alkynyl are optionally substituted by 0-3 R';

[0021] R′ is selected from hydrogen, F, Cl, Br, I, hydroxyl, acyl, carboxyl, amino, nitro, cyano, C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl and C2-C6 alkynyl; the C1-C6 alkyl, C3-C6 cycloalkyl, C2-C6 alkenyl, C2-C6 alkynyl, amino and hydroxyl are optionally substituted with 0-3 hydrogens, hydroxyls, carboxyls, carbonyls, F, Cl, Br, I, amino, nitro, cyano, methyl, trifluoroethyl, difluoromethyl, monofluoromethyl;

[0022] In one or more embodiments of the present application,

[0023] R1 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0024] R2 is selected from hydrogen, C1-C6 alkyl, amino, wherein the amino and C1-C6 alkyl are substituted with 1-3 Ra;

[0025] R a 、R aa are each independently selected from hydrogen, NR b R c 、C1-C6 alkyl or

[0026] R3 is selected from C1-C6 alkyl, C3-C6 cycloalkyl, halogen and -NR b R c ; the C1-C6 alkyl and C3-C6 cycloalkyl are optionally substituted with 0-3 R′;

[0027] R4 is selected from halogen, C1-C6 alkyl and C3-C6 cycloalkyl; the C1-C6 alkyl and C3-C6 cycloalkyl are optionally substituted with 0-3 R′;

[0028] R5 is selected from C1-C6 alkyl and halogen; the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0029] R6 is selected from hydrogen or methyl;

[0030] R b 、R c are each independently selected from hydrogen, C1-C6 alkyl and C2-C6 alkenyl; the C1-C6 alkyl and C2-C6 alkenyl are optionally substituted with 0-3 R′;

[0031] In one or more embodiments of the present application, the compound or its pharmaceutically acceptable salt, wherein

[0032] R3 is selected from C1-C6 alkyl and C3-C6 cycloalkyl; the C1-C6 alkyl and C3-C6 cycloalkyl are optionally substituted with 0-3 R′;

[0033] R4 is selected from halogen and C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R';

[0034] R5 is selected from C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R';

[0035] R6 is hydrogen.

[0036] In one or more embodiments of the present application, the compound or its pharmaceutically acceptable salt, wherein

[0037] R2 is selected from amino and the amino is optionally substituted with 0-3 Ra; wherein, X is selected from CH, N or O;

[0038] R a is selected from C1-C3 alkyl or NR b R c ; wherein Y is selected from N or O, and Z is selected from CH or N;

[0039] R b 、R c are each independently selected from hydrogen, C1-C3 alkyl;

[0040] R3 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R';

[0041] R4 is selected from Cl, Br, C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted with 0-3 R';

[0042] R5 is selected from C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted with 0-3 R';

[0043] R' is selected from hydrogen, C1-C6 alkyl, F, Cl, Br, I, hydroxy and amino.

[0044] In one or more embodiments of the present application, R2 is selected from amino and the amino is optionally substituted with 0-3 Ra; wherein, X is selected from CH, N or O;

[0045] R a is selected from C1-C3 alkyl or NR b R c ;

[0046] wherein Y is selected from N or O, and Z is selected from CH or N;

[0047] R b 、R cEach independently selected from hydrogen, C1-C3 alkyl;

[0048] R3 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R';

[0049] R4 is selected from Cl, Br, C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted with 0-3 R';

[0050] R5 is selected from C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted with 0-3 R'

[0051] R' is selected from F, Cl, Br, I, hydroxy, and amino.

[0052] In one or more embodiments of the present application, R1 is selected from methyl, ethyl, isopropyl, CF2H, CH2CF3;

[0053] R2 is selected from

[0054] R3 is selected from methyl, ethyl, and isopropyl;

[0055] R4 is selected from Cl, Br, CH3, CF3, and CH2CF3;

[0056] R5 is selected from methyl, ethyl, and isopropyl.

[0057] In one or more embodiments of the present application, R1 is selected from methyl, ethyl, isopropyl, CF2H, CH2CF3;

[0058] R2 is selected from

[0059] R3 is selected from methyl and ethyl;

[0060] R4 is selected from Cl, Br, CF3, and CH2CF3;

[0061] R5 is selected from methyl and ethyl.

[0062] In one or more embodiments of the present application, the compound or a pharmaceutically acceptable salt thereof, wherein

[0063] R3 is a halogen;

[0064] R4 is selected from halogen, C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R';

[0065] R5 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R';

[0066] R6 is hydrogen;

[0067] R′ is selected from hydrogen, C1-C6 alkyl, F, Cl, Br, I, hydroxy and amino.

[0068] In one or more embodiments of the present application, R3 is a halogen;

[0069] R4 is selected from halogen, C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0070] R5 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0071] R6 is hydrogen

[0072] R′ is selected from F, Cl, Br, I, hydroxy and amino.

[0073] In one or more embodiments of the present application,

[0074] R1 is selected from methyl, ethyl and isopropyl;

[0075] R2 is selected from

[0076] R3 is selected from Cl and Br;

[0077] R4 is selected from Cl, Br, CF3 and CH2CF3;

[0078] R5 is selected from methyl, ethyl and isopropyl.

[0079] In one or more embodiments of the present application, R1 is selected from methyl, ethyl and isopropyl;

[0080] R2 is selected from

[0081] R3 is selected from Cl and Br;

[0082] R4 is selected from Cl, Br, CF3 and CH2CF3;

[0083] R5 is selected from methyl and ethyl.

[0084] In one or more embodiments of the present application,

[0085] R3 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R′

[0086] R4 is selected from halogen, C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0087] R5 is selected from halogen;

[0088] R6 is hydrogen;

[0089] R′ is selected from hydrogen, C1-C6 alkyl, F, Cl, Br, I, hydroxy, and amino.

[0090] In one or more embodiments of the present application, R3 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R′

[0091] R4 is selected from halogen, C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0092] R5 is selected from halogen;

[0093] R6 is hydrogen;

[0094] R′ is selected from F, Cl, Br, I, hydroxy, and amino.

[0095] In one or more embodiments of the present application,

[0096] R1 is selected from methyl, ethyl, isopropyl;

[0097] R2 is selected from

[0098] R3 is selected from methyl, ethyl, and isopropyl;

[0099] R4 is selected from Cl, Br, CF3, and CH2CF3;

[0100] R5 is selected from F and Cl.

[0101] In one or more embodiments of the present application, R1 is selected from methyl, ethyl, isopropyl;

[0102] R2 is selected from

[0103] R3 is selected from methyl and ethyl;

[0104] R4 is selected from Cl, Br, CF3, and CH2CF3;

[0105] R5 is selected from F and Cl.

[0106] In one or more embodiments of the present application,

[0107] R3 is -NR b R c ;

[0108] R4 is selected from halogen, C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0109] R5 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R′;

[0110] R6 is hydrogen;

[0111] R b 、R c are each independently selected from hydrogen, C1-C6 alkyl, and C2-C6 alkenyl; the C1-C6 alkyl and C2-C6 alkenyl are optionally substituted with 0-3 R';

[0112] R' is selected from hydrogen, C1-C6 alkyl, F, Cl, Br, I, hydroxy, and amino.

[0113] In one or more embodiments of the present application, R3 is -NR b R c ;

[0114] R4 is selected from halogen, C1-C6 alkyl; wherein the C1-C6 alkyl is optionally substituted with 0-3 R';

[0115] R5 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 R';

[0116] R6 is hydrogen;

[0117] R b 、R c are each independently selected from hydrogen, C1-C6 alkyl, and C2-C6 alkenyl; the C1-C6 alkyl and C2-C6 alkenyl are optionally substituted with 0-3 R';

[0118] R' is selected from F, Cl, Br, I, hydroxy, and amino.

[0119] In one or more embodiments of the present application,

[0120] R1 is selected from methyl, ethyl, and isopropyl;

[0121] R2 is selected from

[0122] R3 is NHCH3;

[0123] R4 is selected from Cl, Br, CF3, and CH2CF3;

[0124] R5 is selected from methyl and ethyl.

[0125] In one or more embodiments of the present application,

[0126] R1 is selected from methyl, ethyl, isopropyl, -CF2H;

[0127] R2 is selected from

[0128] R3 is selected from methyl, ethyl, and isopropyl;

[0129] R4 is selected from Br;

[0130] R5 is selected from methyl, ethyl and isopropyl.

[0131] In one or more embodiments of the present application,

[0132] R1 is selected from methyl, ethyl, isopropyl;

[0133] R2 is selected from

[0134] R3 is selected from methyl, ethyl and isopropyl;

[0135] R4 is selected from Br;

[0136] R5 is selected from F, Cl.

[0137] The present invention provides the following compounds or their pharmaceutically acceptable salts:

[0138]

[0139]

[0140]

[0141]

[0142] One or more embodiments of the present application provide a pharmaceutical composition comprising a compound of the present application or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0143] One or more embodiments of the present application provide a pharmaceutical preparation comprising a compound of the present application or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable excipient.

[0144] One or more embodiments of the present application provide the use of a compound of the present application or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical preparation or pharmaceutical composition, in the preparation of a drug for preventing and / or treating diseases related to EGFR or ALK or EGFR and ALK.

[0145] In one or more embodiments, the diseases related to EGFR or ALK or EGFR and ALK are cancers.

[0146] One or more embodiments of the present application provide the use of a compound of the present application or a pharmaceutically acceptable salt thereof, or the pharmaceutical preparation or pharmaceutical composition of the present application, in the preparation of a drug for preventing and / or treating cancer.

[0147] One or more embodiments of the present application provide a compound of the present application or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical preparation or pharmaceutical composition, for use as a medicament.

[0148] One or more embodiments of the present application provide a compound of the present application or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical preparation or pharmaceutical composition, which is used for a method of preventing and / or treating cancer.

[0149] One or more embodiments of the present application provide a compound of the present application or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical preparation or pharmaceutical composition, which is used for preventing and / or treating diseases related to EGFR or ALK or both EGFR and ALK.

[0150] One or more embodiments of the present application provide a compound of the present application or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical preparation or pharmaceutical composition, which is used as an EGFR inhibitor or an ALK inhibitor or an inhibitor of both EGFR and ALK or a protein kinase inhibitor. One or more embodiments of the present application provide the use of a compound of the present application or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical preparation or pharmaceutical composition in the preparation of an EGFR inhibitor.

[0151] One or more embodiments of the present application provide the use of a compound of the present application or a pharmaceutically acceptable salt thereof or the above-mentioned pharmaceutical preparation or pharmaceutical composition in the preparation of an ALK inhibitor.

[0152] One or more embodiments of the present application provide the use of a compound of the present application and its pharmaceutically acceptable salt, or the above-mentioned pharmaceutical preparation or pharmaceutical composition in the preparation of an inhibitor of both EGFR and ALK.

[0153] In one or more embodiments of the present application, the drug for treating cancer is an arylphosphoryl compound having protein kinase inhibitor activity, including: drugs for preventing and / or treating lung cancer, such as drugs for preventing and / or treating multiple myeloma; and drugs for preventing and / or treating lymphoma, such as drugs for preventing and / or treating non-Hodgkin lymphoma, mantle cell lymphoma and / or follicular lymphoma; drugs for preventing and / or treating leukemia; and drugs for preventing and treating mantle cell tumor, breast cancer, liver cancer, colon cancer, cervical cancer, lung cancer, plasmacytoma, lymphoma, ovarian cancer, kidney cancer, gastric cancer, nasopharyngeal cancer, leukemia, melanoma, thyroid cancer, pancreatic cancer, adenocarcinoma or squamous cell carcinoma.

[0154] In one or more embodiments of the present application, the compound of the present application can be used to treat lung cancer, plasmacytoma, mantle cell tumor, multiple myeloma, melanoma, breast cancer, liver cancer, cervical cancer, lymphoma, leukemia, ovarian cancer, kidney cancer, gastric cancer, nasopharyngeal cancer, thyroid cancer, pancreatic cancer, prostate cancer, adenocarcinoma, oral cancer, esophageal cancer, squamous cell carcinoma or colon cancer.

[0155] One or more embodiments of the present application also provide a method for treating and / or preventing diseases associated with EGFR or ALK or both EGFR and ALK, the method comprising administering to a subject in need thereof a compound of the present application or a pharmaceutically acceptable salt thereof, or a pharmaceutical preparation or pharmaceutical composition of the present application.

[0156] One or more embodiments of the present application also provide a method for treating and / or preventing cancer or tumor, the method comprising administering to a subject in need thereof a compound of the present application or a pharmaceutically acceptable salt thereof, or a pharmaceutical preparation or pharmaceutical composition of the present application.

[0157] One or more embodiments of the present application also provide the use of a compound of the present application or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical preparation or pharmaceutical composition in the preparation of a protein kinase inhibitor.

[0158] One or more embodiments of the present application also provide a method for inhibiting an EGFR inhibitor and / or an ALK inhibitor or a protein kinase in vivo or in vitro, the method comprising administering to a subject or a subject in need thereof a compound of the present application or a pharmaceutically acceptable salt thereof, or the above-mentioned pharmaceutical preparation or pharmaceutical composition.

[0159] The following explains the terms used in the technical solutions of the present application. As used in the specification and the appended claims, unless otherwise specifically stated to the contrary, the terms of the present application have the meanings indicated as follows:

[0160] The term "compound" includes all stereoisomers, geometric isomers, and tautomers. The "compound" described herein may be asymmetric, for example, having one or more stereoisomers. Unless otherwise specified, all stereoisomers are included, such as individual enantiomers and diastereomers or other stereoisomeric forms or mixtures thereof. Compounds containing asymmetric carbon atoms herein may be isolated in optically pure form or as a racemate. The optically pure form can be resolved from a racemic mixture or synthesized by using chiral starting materials or chiral reagents. The "compound" described herein also includes geometric isomeric forms, which refer to forms of a compound in which the substituents on a double bond or a ring have different cis-trans isomers without chirality. The "compound" described herein also includes tautomeric forms. Tautomeric forms can result from the exchange of a single bond with an adjacent double bond and the accompanying migration of a proton.

[0161] The compounds of the present invention, whether intermediates or compounds of formula (I), may also be isotopically labeled by replacing one or more atoms therein with atoms having different atomic masses or mass numbers. Such isotopically labeled (i.e., radiolabeled) compounds are considered to be within the scope of the present invention. Examples of isotopes in the compounds of the present invention include isotopes of carbon, nitrogen, oxygen, sulfur, fluorine, chlorine, and iodine, each having the same number of protons but different mass numbers.

[0162] The term "halogen" refers to fluorine, chlorine, bromine, or iodine.

[0163] The term "amino" refers to -NH2.

[0164] The term "cyano" refers to -CN.

[0165] The term "nitro" refers to -NO2.

[0166] The term "hydroxy" refers to -OH.

[0167] The term "carboxy" refers to -COOH.

[0168] The term "alkyl" refers to a saturated aliphatic hydrocarbon group, and this term includes straight-chain and branched-chain hydrocarbon groups. For example, C1-C 20 alkyl, preferably C1-C6 alkyl. C1-C 20 alkyl refers to an alkyl group having 1 to 20 carbon atoms, such as an alkyl group having 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, 10 carbon atoms, 11 carbon atoms, 12 carbon atoms, 13 carbon atoms, 14 carbon atoms, 15 carbon atoms, 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, 19 carbon atoms, or 20 carbon atoms. Non-limiting examples of alkyl include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, neopentyl, n-hexyl, etc. The alkyl may be unsubstituted or substituted by one or more substituents, and the substituents include but are not limited to alkyl, alkoxy, cyano, hydroxy, carbonyl, carboxy, aryl, heteroaryl, amino, halogen, sulfonyl, sulfinyl, phosphonyl, etc.

[0169] The term "cycloalkyl" refers to a cyclic alkyl having a monocyclic or polycyclic (including fused rings, bridged rings, and spiro ring systems), including a cyclic alkyl composed of 3 to 8 carbon atoms (for example, 3, 4, 5, 6, 7, or 8 carbon atoms) and hydrogen atoms. Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, spiro[3.4]octyl, bicyclo[1.1.1]pentyl, bicyclo[3.1.0]hexyl, etc.

[0170] The term "cycloalkenyl" refers to a cyclic or polycyclic hydrocarbon group having 3 to 13 carbon atoms, preferably 5 to 8 carbon atoms, and containing one or more double bonds. The cycloalkenyl group may be substituted or unsubstituted. The cycloalkenyl group includes, but is not limited to, cyclopentenyl, cyclohexenyl, and cyclooctenyl.

[0171] The term "alkenyl" refers to a hydrocarbon group containing one or more double bonds in a straight-chain or branched-chain hydrocarbon chain. The alkenyl group may be unsubstituted or substituted. The alkenyl group may have 1 to 20 carbon atoms, and a numerical range such as "1 to 20" means each integer within the given range; for example, "1 to 20 carbon atoms" means an alkenyl group that may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, 10 carbon atoms, 11 carbon atoms, 12 carbon atoms, 13 carbon atoms, 14 carbon atoms, 15 carbon atoms, 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, 19 carbon atoms, or 20 carbon atoms.

[0172] The term "alkynyl" refers to a hydrocarbon group containing one or more triple bonds in a straight-chain or branched-chain hydrocarbon chain. The alkynyl group may be unsubstituted or substituted. The alkynyl group may have 1 to 20 carbon atoms, and a numerical range such as "1 to 20" means each integer within the given range; for example, "1 to 20 carbon atoms" means an alkynyl group that may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 8 carbon atoms, 9 carbon atoms, 10 carbon atoms, 11 carbon atoms, 12 carbon atoms, 13 carbon atoms, 14 carbon atoms, 15 carbon atoms, 16 carbon atoms, 17 carbon atoms, 18 carbon atoms, 19 carbon atoms, or 20 carbon atoms.

[0173] The term "heteroaryl" refers to a monocyclic or fused ring having 5 - 12 ring atoms (e.g., 5, 6, 10, 12, 14 ring atoms), containing 1 - 4 (e.g., 1, 2, 3, or 4) heteroatoms selected from N, O, S, with the remaining ring atoms being C, and having a fully conjugated π - electron system, including but not limited to pyrrolyl, furyl, thienyl, imidazolyl, oxazolyl, isoxazolyl, pyrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, quinolinyl, isoquinolinyl, triazolyl, benzimidazole, benzotriazole, etc. The heteroaryl can be unsubstituted or substituted, and the substituents include but are not limited to alkyl, alkoxy, cyano, hydroxy, carbonyl, carboxyl, aryl, aralkyl, amino, halogen, sulfonyl, sulfinyl, phosphonyl. The term "substituted" as used herein means that any group is mono - or polysubstituted by the specified substituents to the extent that such mono - or polysubstitution (including multiple substitutions at the same moiety) is chemically permissible, each substituent can be located at any available position on the group, and can be attached through any available atom on the substituent. "Any available position" means any position on the group that can be chemically obtained by methods known in the art or taught herein and that does not result in an unduly unstable molecule. When there are two or more substituents on any group, each substituent is defined independently of any other substituent and thus can be the same or different.

[0174] The term "acyl" refers to hydrogen, alkyl, alkenyl, alkynyl, or aryl attached as a substituent through a carbonyl group. Examples include formyl, acetyl, propionyl, benzoyl, and acryloyl. The acyl group can be substituted or unsubstituted.

[0175] The term "substituted" as used herein means that any group is mono - or polysubstituted by the specified substituents to the extent that such mono - or polysubstitution (including multiple substitutions at the same moiety) is chemically permissible, each substituent can be located at any available position on the group, and can be attached through any available atom on the substituent. "Any available position" means any position on the group that can be chemically obtained by methods known in the art or taught herein and that does not result in an unduly unstable molecule. When there are two or more substituents on any group, each substituent is defined independently of any other substituent and thus can be the same or different.

[0176] When a group is described as "optionally substituted by 0 - 6 R′", it means that it can be arbitrarily substituted by 1, 2, 3, 4, 5, or 6 R′, or not substituted by R′, i.e., corresponding to 0 R′. When a group is described as "optionally substituted by 0 - 3 R′", it means that it can be arbitrarily substituted by 1, 2, or 3 R′, or not substituted by R′, i.e., corresponding to 0 R′. When a group is described as "optionally substituted by 0 - 3 Ra When substituted, it means that it can be arbitrarily substituted by one, two or three Rs a substituted, or not substituted by R a substituted, that is, corresponding to 0 Rs a .

[0177] When a group is described as "optionally substituted", the group can be unsubstituted or substituted by one or more of the following substituents: hydrogen, fluorine, chlorine, bromine, iodine, nitro, trifluoromethyl, difluoromethyl, trifluoromethoxy, difluoromethoxy, methoxy, acyl, alkoxy, heterocycloalkyl, alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, cycloalkynyl, aryl, heteroaryl, heteroalicyclic, aralkyl, heteroaralkyl, (heteroalicyclic)alkyl, hydroxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halogen, thiocarbonyl, O-carbamoyl, N-carbamoyl, O-thiocarbamoyl, N-thiocarbamoyl, C-acylamino, N-acylamino, S-sulfonylamino, N-sulfonylamino, carboxyl, isocyanato, thiocyanato, isothiocyanato, silyl, hydrocarbonthio, sulfinyl, sulfonyl, haloalkyl, haloalkoxy and amino.

[0178] The term "pharmaceutically acceptable" means that a substance or composition must be chemically and / or toxicologically compatible with the other components of the formulation and / or the mammal being treated therewith.

[0179] The "pharmaceutical preparation" referred to in the present application can be the drug composition described in the application directly, or in combination with other active ingredients, and combined with pharmaceutically acceptable excipients or carriers. The preparations include: tablets, pills, capsules, granules, suspensions and emulsions, etc. The pharmaceutically acceptable excipients or carriers include: binders such as microcrystalline cellulose, tragacanth or gelatin; excipients such as starch or lactose; dispersants such as alginic acid, Primogel or corn starch; lubricants such as magnesium stearate; glidants such as colloidal silica; sweeteners such as sucrose or saccharin; or flavoring agents such as peppermint oil, methyl salicylate or orange flavor; non-aqueous solvents such as dimethyl sulfoxide, alcohols, propylene glycol, polyethylene glycol, vegetable oils such as olive oil and injectable organic esters such as ethyl oleate; aqueous carriers such as mixtures of alcohols and water, buffered media and saline; and preservatives, antibacterial agents, antioxidants, chelating agents, dyes, pigments or fragrances, etc.

[0180] In one or more embodiments of the present application, the cancer specifically is any one or more of plasmacytoma, mantle cell lymphoma, multiple myeloma, melanoma, breast cancer, liver cancer, cervical cancer, lung cancer, lymphoma, leukemia, ovarian cancer, kidney cancer, gastric cancer, nasopharyngeal cancer, thyroid cancer, pancreatic cancer, prostate cancer, adenocarcinoma, oral cancer, esophageal cancer, squamous cell carcinoma or colon cancer.

[0181] Other anti-tumor drugs that can be used together with the compounds of the present application to form a pharmaceutical composition include: cytotoxic drugs, hormone drugs, antimetabolites, tumor-targeted drugs, and adjuvant therapy drugs, etc. Cytotoxic drugs, such as carboplatin, cisplatin, irinotecan, paclitaxel, fluorouracil, cytarabine, lenalidomide, and retinoic acid; Hormone drugs, such as dexamethasone, fulvestrant, tamoxifen, etc.; Antimetabolites, fluorouracil, methotrexate, furanfluorouracil, cytarabine; Molecular targeted drugs, such as imatinib, erlotinib, lapatinib, etc. of tinib drugs, and PARP inhibitor drugs, such as Olaparib, Rubraca, Zejula, etc.; Adjuvant therapy drugs, such as recombinant human granulocyte colony stimulating factor, erythropoietin, pamidronate disodium, zoledronic acid, etc. In addition, it also includes anti-tumor biological drugs such as Keytruda, Opdiv, Tecentriq, Imfinzi, Bavencio, etc.

[0182] Technical Effects

[0183] The compounds in one or more embodiments of the present invention can effectively inhibit various types of EGFR drug-resistant mutations (such as EGFR del19 , EGFR del19 / T790M , EGFR del19 / C797S , EGFR T790M / L858R , EGFR L858R / C797S , EGFR del19 / T790M / C797S , EGFR L858R / T790M / C797S ) activity, and the compounds in one or more embodiments of the present invention have good inhibitory effects on ALK fusion genes and mutations thereof. The compounds in one or more embodiments of the present invention have inhibitory activity on EGFR drug-resistant mutations and ALK fusion genes and mutations thereof.

[0184] In one or more embodiments, for various EGFR drug-resistant mutations characterized by T790M mutation, the IC values ​​of the compounds of the present invention are higher than those of the compounds disclosed in the prior art. 50 Value and / or GI 50 The value was significantly reduced, showing better inhibitory activity; a variety of EGFR resistance mutations with T790M mutation characteristics, including EGFR-T790M / Del19, EGFR-T790M / L858R, EGFR-C797S / T790M / L858R, EGFR-C797S / T790M / Del19, etc.

[0185] In one or more embodiments, for various EGFR drug-resistant mutations characterized by C797S mutation, the IC values ​​of the compounds of the present invention are higher than those of the compounds disclosed in the prior art. 50 Value and / or GI 50The value is significantly reduced, showing better inhibitory activity; various EGFR drug-resistant mutations with the C797S mutation feature, including EGFR-C797S / Del19, EGFR-C797S / L858R, EGFR-C797S / T790M / L858R, EGFR-C797S / T790M / Del19, etc.

[0186] In one or more embodiments, for various EGFR drug-resistant mutations with the L858R mutation feature, the IC 50 value and / or GI 50 value of the compounds of the present invention are significantly reduced compared to the compounds disclosed in the prior art, showing better inhibitory activity. For various EGFR drug-resistant mutations with the Del19 mutation feature, the IC 50 value and / or GI 50 value of the compounds of the present invention are significantly reduced compared to the compounds disclosed in the prior art, showing better inhibitory activity.

[0187] In one or more embodiments, for mutant EGFR and wild-type EGFR, the compounds in one or more embodiments of the present invention also show higher selectivity compared to the compounds disclosed in the prior art.

[0188] In one or more embodiments, for ALK mutant genes, such as EML4-ALK, EML4-ALK-L1196M, etc., the IC 50 value and / or GI 50 value of the compounds in one or more embodiments of the present invention are significantly reduced compared to the compounds disclosed in the prior art, showing better inhibitory activity.

[0189] In one or more embodiments, the compounds of the present application show an unexpected activity effect in the inhibitory EGFR drug-resistant mutation activity test compared to the compounds of the comparative examples, and at the same time also show significant and unexpected inhibitory activity against ALK fusion genes and mutants.

[0190] In one or more embodiments, the compounds of the present application show higher safety and a significantly increased tolerance dose compared to the compounds of the comparative examples.

[0191] The compounds of one or more embodiments of the present application have good physicochemical properties and high stability. Detailed Description of the Invention

[0192] The present application will be further described below in conjunction with embodiments, providing the implementation details of the present application. It should be noted that the embodiments described below are exemplary and are only used to explain the present application, and should not be construed as a limitation to the present application. Modifications or substitutions made by those skilled in the art based on the prior art still fall within the protection scope of the present application. The reagents used in the embodiments of the present application can all be obtained through commercial purchases.

[0193] Example 1

[0194] Compound 1A:

[0195]

[0196] In a 500 mL three-necked flask, 2-iodo-4-methylaniline (10 g, 42.9 mmol), K3PO4 (10.9 g, 51.5 mmol), Xantphos (2.48 g, 4.3 mmol), Pd(OAc)2 (0.96 g, 4.3 mmol), dimethylphosphite (5 g, 64.4 mmol), and 100 mL of DMF were added. The temperature was raised to 120 °C and the reaction was carried out for about 3 h until the reaction was completed as detected by TLC. The reaction solution was cooled to room temperature, filtered by suction, 600 mL of H2O was added, a large amount of yellow solid was precipitated, filtered by suction, the filtrate was extracted with 500 mL × 3 of ethyl acetate, the organic phases were combined, washed with 250 mL × 2 of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain Compound 1A, which was used in the next step without purification. 1 1H NMR (400 MHz, DMSO-d6) δ = 9.21 (brs, 2H), 7.57 (d, J = 13.6, 1H), 7.43 (d, J = 8.4, 1H), 7.26 - 7.23 (m, 1H), 2.33 (s, 3H), 1.85 (s, 3H), 1.82 (s, 3H).

[0197] Compound 1B:

[0198]

[0199] In a 500 mL three-necked flask, the Compound 1A obtained in the previous step, 2,4,5-trichloropyrimidine (11.80 g, 64.3 mmol), K2CO3 (7.76 g, 128.73 mmol), nBu4NHSO4 (1.45 g, 4.29 mmol), and 100 mL of DMF were added. The temperature was raised to 65 °C and the reaction was carried out for about 4.5 h until the reaction was completed as detected by TLC. The reaction was cooled to room temperature, 200 mL of H2O was added, a large amount of yellow solid was precipitated, and after stirring for about 0.5 h, it was filtered by suction. The filter cake was washed with 100 mL of H2O and dried to obtain Compound 1B (8.57 g, total two-step yield 60.5%). 1HNMR (400 MHz, DMSO) δ = 11.62 (s, 1H), 8.40 (s, 1H), 8.27 (dd, J = 8.4, 4.4 Hz, 1H), 7.44 (m, 2H), 2.33 (s, 3H), 1.81 (s, 3H), 1.77 (s, 3H).

[0200] Compound 1C:

[0201]

[0202] In a 250 mL three-necked flask, weigh 3-fluoro-4-methylphenol (10 g, 79.28 mmol), benzyltriethylammonium chloride (1.77 g, 7.93 mmol), add 80 mL of dichloromethane and stir to dissolve. Slowly add nitric acid (14.7 g, 158.57 mmol) dropwise to the above reaction solution at 0 - 10 °C. After the addition, continue to stir the reaction mixture for about 0.5 h, and detect the completion of the reaction by TLC. Then slowly add 120 mL of saturated sodium bicarbonate aqueous solution, separate the layers, and the organic phase is successively washed with 40 mL of H2O and 40 mL of saturated sodium chloride aqueous solution.

[0203] The organic phase is concentrated and purified by column chromatography (PE / EA = 8 / 1) to obtain Compound 1C (8 g, yield 59.2%). 1 H NMR (400 MHz, CDCl3) δ = 10.66 (d, J = 0.8 Hz, 1H), 8.02 (d, J = 7.6 Hz, 1H), 6.83 (d, J = 10.0 Hz, 1H), 2.28 (d, J = 0.8 Hz, 3H).

[0204] Compound 1D:

[0205]

[0206] In a 100 mL three-necked flask, weigh Compound 1C (2 g, 11.68 mmol), K2CO3 (2.42 g, 17.54 mmol), methyl iodide (1.08 mL, 17.54 mmol), add 30 mL of DMF, stir at room temperature overnight, and detect the completion of the reaction by TLC. Then add 60 mL of water to the reaction solution, a large amount of off-white solid precipitates, continue to stir for about 0.5 h and then filter by suction. The filter cake is washed with water and dried to obtain Compound 1D (1.59 g, yield 73.6%). 1 H NMR (400 MHz, CDCl3) δ = 7.85 (d, J = 7.6 Hz, 1H), 6.79 (d, J = 11.2 Hz, 1H), 3.05 (s, 3H), 2.27 (d, J = 1.6 Hz, 3H).

[0207] Compound 1E:

[0208]

[0209] In a 250 mL three-necked flask, weigh in compound 1D (4.5 g, 24.3 mmol), K2CO3 (6.71 g, 48.6 mmol), 1-methyl-4-(4-piperidyl)piperazine (6.67 g, 36.4 mmol), and 67.5 mL of DMF. Heat to 120 °C and stir the reaction for about 4 hours until the reaction is complete as detected by TLC. Then cool to room temperature, add 130 mL of H2O to the reaction solution, a large amount of solid precipitates, continue to stir for about 0.5 hours and then filter by suction. Wash the filter cake with water and dry to obtain compound 1E (4.82 g, yield 56.8%). MS-ESI (m / z): 349.2248 (M+H) + 。

[0210] Compound 1F:

[0211]

[0212] In a 250 mL single-necked flask, weigh in compound 1E (4.5 g, 12.91 mmol), 0.45 g of 5% Pd / C, and 90 mL of methanol. Replace the gas with hydrogen 2 - 3 times, stir at room temperature overnight under a hydrogen atmosphere (atmospheric pressure) until the reaction is complete as detected by TLC. Then filter through diatomaceous earth, concentrate the filtrate under vacuum, and purify by column chromatography (DCM / MeOH = 8 / 1) to obtain compound 1F (4.02 g, yield 97.8%). MS-ESI (m / z): 319.2509 (M+H) + 。 1 1H NMR (400 MHz, CDCl3) δ = 6.57 (d, J = 4.2 Hz, 2H), 3.83 (s, 3H), 3.08 (d, J = 8.4 Hz, 2H), 2.81 - 2.45 (m, 10H), 2.32 (s, 4H), 2.17 (s, 3H), 2.01 - 1.87 (m, 2H), 1.70 - 1.68 (m, 2H). MS-ESI (m / z): 319.2509 (M+H) + 。

[0213] Compound 1:

[0214]

[0215] Compound 1B (500 mg, 1.51 mmol), compound 1F (667 mg, 2.12 mmol), ethanol solution of 15% hydrogen chloride (960 mg, 3.64 mmol), and 7.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 15 mL of saturated aqueous NaHCO3 solution was added, and a large amount of solid was precipitated. After stirring for an additional 0.5 h, filtration was performed. The filter cake was washed with water, and the filter cake was purified by slurrying with 9 mL of EtOH / H2O = 1 / 2 mixed solvent to obtain compound 1 (707 mg, yield 76.3%). 1 1H NMR (400 MHz, CDCl3) δ = 10.57 (s, 1H), 8.45 (dd, J = 8.4, 4.8 Hz, 1H), 8.05 (m, 2H), 7.33 (m, 2H), 7.08 (dd, J = 14.4, 1.2 Hz, 1H), 6.62 (s, 1H), 3.86 (s, 3H), 3.15 (d, J = 11.6 Hz, 2H), 2.81 - 2.59 (m, 6H), 2.51 (m, 2H), 2.37 (s, 3H), 2.31 (s, 3H), 2.18 (s, 3H), 2.12 (s, 3H), 1.96 (m, 2H), 1.85 (s, 3H), 1.82 (s, 3H), 1.72 (m, 2H). MS-ESI (m / z): 612.3098 (M + H) + 。

[0216] Example 2

[0217] Compound 2B:

[0218]

[0219] In a 250 mL three-necked flask, compound 1A (3.00 g, 16.38 mmol), 5-bromo-2,4-dichloropyrimidine (5.6 g, 24.57 mmol), K2CO3 (6.79 g, 49.14 mmol), nBu4NHSO4 (0.56 g, 1.638 mmol), and 60 mL of DMF were added. The temperature was raised to 65 °C and the reaction was carried out for about 4.5 h until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 200 mL of H2O was added, and a large amount of yellow solid was precipitated. After stirring for about 0.5 h, filtration was performed. The filter cake was washed with 100 mL of H2O and dried to obtain compound 2B (4.5 g, yield 73.4%). 11H NMR (400 MHz, CDCl3) δ = 11.19 (s, 1H), 8.43 (dd, J = 8.4, 4.8 Hz, 1H), 8.32 (s, 1H), 7.41 (d, J = 8.8 Hz, 1H), 7.08 (m, 1H), 2.38 (s, 3H), 1.86 (s, 3H), 1.83 (s, 3H). MS-ESI (m / z): 395.9642 (M+Na) + 。

[0220] Compound 2:

[0221]

[0222] Compound 2B (375 mg, 1.00 mmol), compound 1F (414 mg, 1.30 mmol), ethanol solution of 15% hydrogen chloride (730 mg, 3.00 mmol), and 4.0 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 50 mL of 3% K2CO3 aqueous solution and 50 mL of dichloromethane were added. After stirring for 10 minutes, the mixture was allowed to stand for liquid separation. The upper aqueous phase was washed with 50 mL of dichloromethane once more. The dichloromethane phases from the two extractions were combined. Then, the combined dichloromethane phase was washed with 50 mL of saturated brine, dried over anhydrous sodium sulfate, filtered, the filtrate was concentrated, and after purification by column chromatography, compound 2 (360 mg, yield 54.83%) was obtained. 1 1H NMR (400 MHz, CDCl3) δ = 10.29 (s, 1H), 8.32 (dd, J = 8.8, 4.8 Hz, 1H), 8.18 (s, 1H), 8.01 (s, 1H), 7.44 - 7.30 (m, 2H), 7.11 (d, J = 14.0 Hz, 1H), 6.61 (s, 1H), 3.85 (s, 3H), 3.15 (d, J = 11.6 Hz, 2H), 2.80 - 2.52 (m, 11H), 2.38 (s, 3H), 2.35 (s, 3H), 2.14 (s, 3H), 1.96 (d, J = 11.0 Hz, 2H), 1.85 (s, 3H), 1.82 (s, 3H), 1.78 - 1.68 (m, 2H). MS-ESI (m / z): 656.2459 (M+H) + 。

[0223] Example 3

[0224] Compound 3B:

[0225]

[0226] In a 50 mL three-necked flask, add compound 1A (420 mg, 2.29 mmol), 5-trifluoromethyl-2,4-dichloropyrimidine (745 mg, 3.43 mmol), K2CO3 (949 mg, 6.87 mmol), nBu4NHSO4 (78 mg, 0.23 mmol), and 8.5 mL of DMF. Heat the mixture to 65 °C and react for about 4.5 hours until the reaction is completed as detected by TLC. Cool the reaction to room temperature, add 18.5 mL of H2O, and a large amount of yellow solid precipitates. Continue stirring for about 0.5 h and then filter by suction. Wash the filter cake with 20 mL of H2O and dry to obtain compound 3B (466 mg, yield 56.0%). 1 1H NMR (400 MHz, CDCl3) δ = 11.49 (s, 1H), 8.58 (s, 1H), 8.50 - 8.47 (dd, J = 8.4 Hz, 4.4 Hz, 1H), 7.39 (d, J = 8.4 Hz, 1H), 7.08 - 7.04 (dd, J = 14.4 Hz, 1.2 Hz, 1H), 2.37 (s, 3H), 1.87 (s, 3H), 1.84 (s, 3H). MS-ESI (m / z): 364.0593 (M + H) + ; 386.0413 (M + Na) + .

[0227] Compound 3:

[0228]

[0229] Add compound 3B (181 mg, 0.50 mmol), compound 1F (207 mg, 0.65 mmol), ethanol solution of 15% hydrogen chloride (365 mg, 1.50 mmol), and 3.0 mL of ethylene glycol monomethyl ether to the reaction flask. Then, seal the tube and react at 120 °C for 5 - 6 h. After detecting the completion of the reaction by TLC, cool the reaction solution to room temperature. Then, add 50 mL of 3% K2CO3 aqueous solution and 50 mL of dichloromethane. Stir for 10 minutes and then let it stand for liquid separation. Wash the upper aqueous phase with 50 mL of dichloromethane again. Combine the two dichloromethane phases. Then, wash the dichloromethane phase with 50 mL of saturated brine, dry it over anhydrous sodium sulfate, filter, concentrate the filtrate, and purify it by column chromatography to obtain compound 3 (70 mg, yield 21.68%). 11H NMR (400 MHz, CDCl3) δ = 10.51 (s, 1H), 8.30 (s, 2H), 8.10 (s, 1H), 7.50 (s, 1H), 7.22 (d, J = 8.4 Hz, 1H), 7.07 (d, J = 14.0 Hz, 1H), 6.64 (s, 1H), 3.87 (s, 3H), 3.19 (d, J = 10.4 Hz, 2H), 3.08 - 2.12 (m, 20H), 1.99 (d, J = 10.8 Hz, 2H), 1.79 (m, 8H). MS-ESI (m / z): 646.3213 (M+H) + 。

[0230] Example 4

[0231] Compound 4B:

[0232]

[0233] Add the mixed solvent of DMF / DMSO (15 ml / 1.5 ml) to a 50 mL single-necked flask, cool it in an ice bath, add NaH (0.66 g, 16.38 mmol) to the above mixed solvent in batches. After stirring for 5 - 10 min, add dropwise the mixed solution of compound 1A (1.0 g, 5.46 mmol) in DMF / DMSO (9 ml / 1 ml) from Example 1. After the addition is complete, keep stirring at low temperature for 30 min, then add dropwise the mixed solution of 5-methyl-2,4-dichloropyrimidine (1.33 g, 8.16 mmol) in DMF / DMSO (9 ml / 1 ml). After the addition is complete, slowly raise the temperature to room temperature and react overnight. After detecting the reaction by TLC, add 90 mL of H2O, add 25 ml of ethyl acetate for extraction, extract the aqueous phase with 25 ml of ethyl acetate twice, combine the organic phases, then wash with 50 mL of water twice and 50 ml of saturated brine twice, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and purify by column chromatography to obtain compound 4B (1.2 g, yield 70.59%).

[0234] 1 1H NMR (400 MHz, CDCl3) δ = 10.85 (s, 1H), 8.62 (dd, J = 8.8, 4.8 Hz, 1H), 7.99 (s, 1H), 7.39 (d, J = 8.8 Hz, 1H), 7.03 (m, 1H), 2.36 (s, 3H), 2.24 (s, 3H), 1.85 (s, 3H), 1.82 (s, 3H). MS-ESI (m / z): 310.0874 (M+H) + 。

[0235] Compound 4:

[0236]

[0237] Compound 4B (500 mg, 1.61 mmol), compound 1F (461 mg, 1.45 mmol), ethanol solution of 15% hydrogen chloride (1.18 g, 4.83 mmol), and 7.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 100 °C for 5 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 15 mL of saturated aqueous NaHCO3 solution and 10 mL of water were added. 10 mL of dichloromethane was added and stirred for extraction. The aqueous phase was extracted with dichloromethane twice more. The organic phases were combined, washed twice with 15 mL of water and twice with 15 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain an oily substance. After purification by trituration with a mixed solvent of EA / PE = 3 / 2, compound 4 (120 mg, yield 14.0%) was obtained. 1 1H NMR (400 MHz, CDCl3) δ = 10.17 (s, 1H), 8.59 (dd, J = 8.4, 4.8 Hz, 1H), 8.14 (s, 1H), 7.91 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.26 (s, 1H), 7.04 (d, J = 13.4 Hz, 1H), 6.63 (s, 1H), 3.86 (s, 3H), 3.16 (d, J = 12.0 Hz, 2H), 2.67 (m, 9H), 2.36 (m, 8H), 2.20 (d, J = 6.4 Hz, 6H), 1.97 (m, 2H), 1.84 (s, 3H), 1.81 (s, 3H), 1.79 - 1.69 (m, 2H). MS-ESI (m / z): 592.3526 (M+H) + 。

[0238] Example 5

[0239] Compound 5E:

[0240]

[0241] In a 100 mL single-necked flask, compound 1D (1.5 g, 8.10 mmol), K2CO3 (2.24 g, 16.20 mmol), 4-dimethylaminopiperidine (1.56 g, 12.15 mmol), and 30 mL of DMF were weighed. The temperature was raised to 120 °C and stirred for about 4 hours until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 60 mL of H2O was added, and a large amount of solid precipitated. Stirring was continued for about 0.5 hours and then filtered by suction. The filter cake was washed with water and dried to obtain compound 5E (1.46 g, yield 61.3%). MS-ESI (m / z): 294.1825 (M+H) + 。

[0242] Compound 5F:

[0243]

[0244] In a 250 mL single-necked flask, weigh in compound 5E (1.4 g, 4.77 mmol), 0.52 g of 5% Pd / C, and 28 mL of methanol. Replace the air with hydrogen 2 - 3 times. Stir at room temperature overnight under a hydrogen atmosphere (atmospheric pressure) until the reaction is complete as detected by TLC. After filtration through diatomaceous earth, the filtrate is concentrated in vacuo and purified by column chromatography (DCM / MeOH = 8 / 1) to obtain compound 5F (918 mg, yield 73.0%). MS-ESI (m / z): 264.2065 (M + H) + 。

[0245] Compound 5:

[0246]

[0247] Add compound 1B (400 mg, 1.21 mmol), compound 5F (383 mg, 1.45 mmol), ethanol solution of 15% hydrogen chloride (884 mg, 3.63 mmol), and 6.0 mL of ethylene glycol monomethyl ether to the reaction flask. Seal the tube and react at 120 °C for 5 - 6 h until the reaction is complete as detected by TLC. Subsequently, cool the reaction solution to room temperature, add 15 mL of saturated aqueous NaHCO3 and 10 mL of H2O. A large amount of solid precipitates. Continue to stir for 0.5 h and then filter by suction. Wash the filter cake with water and purify it by pulping with 6 mL of a mixed solvent of PE / EA = 5 / 1 to obtain compound 5 (410 mg, yield 61.5%). 1 1H NMR (400 MHz, CDCl3) δ = 10.58 (s, 1H), 8.45 (dd, J = 8.4, 4.8 Hz, 1H), 8.06 (m, 2H), 7.33 (m, 2H), 7.09 (d, J = 14.4 Hz, 1H), 6.64 (s, 1H), 3.86 (s, 3H), 3.15 (d, J = 12.0 Hz, 2H), 2.65 (m, 2H), 2.37 (m, 9H), 2.27 (m, 1H), 2.19 (s, 3H), 1.93 (m, 2H), 1.85 (s, 3H), 1.82 (s, 3H), 1.70 (m, 2H). MS-ESI (m / z): 557.2571 (M + H) + 。

[0248] Example 6

[0249] Compound 6:

[0250]

[0251] Compound 2B (400 mg, 1.07 mmol), compound 5F (422 mg, 1.60 mmol), ethanol solution of 15% hydrogen chloride (779 mg, 3.20 mmol), and 6.0 mL of ethylene glycol monomethyl ether were added to a reaction flask, and the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 15 mL of saturated aqueous NaHCO3 solution and 10 mL of H2O were added, a large amount of solid was precipitated, and the mixture was stirred for another 0.5 h and then filtered. The filter cake was washed with water, dried, and separated by column chromatography (DCM / MeOH = 8 / 1). After purification, it was slurried with 5 mL of a mixed solvent of PE / EA = 4 / 1 to obtain compound 6 (276 mg, yield 43.0%). 1 1H NMR (400 MHz, CDCl3) δ = 10.30 (s, 1H), 8.33 (dd, J = 8.4, 4.4 Hz, 1H), 8.18 (s, 1H), 8.03 (s, 1H), 7.43 - 7.30 (m, 2H), 7.11 (d, J = 14.0 Hz, 1H), 6.62 (s, 1H), 3.86 (s, 3H), 3.16 (d, J = 11.8 Hz, 2H), 2.65 (m, 2H), 2.40 (m, 10H), 2.14 (s, 3H), 2.03 - 1.94 (m, 2H), 1.85 (s, 3H), 1.82 (s, 3H), 1.73 (m, 2H). MS-ESI (m / z): 601.2015 (M+H) + 。

[0252] Example 7

[0253] Compound 7:

[0254]

[0255] Referring to the synthesis method of compound 5, only compound 1B was replaced with compound 3B to obtain compound 7. MS-ESI (m / z): 591.2856 (M+H) + 。 1 1H NMR (400 MHz, CDCl3) δ = 10.55 (s, 1H), 8.36 (s, 2H), 8.12 (s, 1H), 7.52 (s, 1H), 7.25 (d, J = 8.4 Hz, 1H), 7.10 (d, J = 14.0 Hz, 1H), 6.65 (s, 1H), 3.82 (s, 3H), 3.13 (d, J = 12.0 Hz, 2H), 2.65 (m, 2H), 2.38 (m, 10H), 2.17 (s, 3H), 1.95 (m, 2H), 1.86 (s, 3H), 1.83 (s, 3H), 1.71 (m, 2H).

[0256] Example 8

[0257] Compound 8:

[0258]

[0259] Referring to the synthesis method of Reference Compound 5, only Compound 1B therein was replaced with Compound 4B to obtain Compound 8. MS-ESI (m / z): 537.3169 (M+H) + 。 1 H NMR (400 MHz, CDCl3) δ = 10.25 (s, 1H), 8.45 (dd, J = 8.4, 4.8 Hz, 1H), 8.12 (s, 1H), 7.89 (s, 1H), 7.35 (m, 2H), 7.04 (d, J = 14.0 Hz, 1H), 6.60 (s, 1H), 3.83 (s, 3H), 3.15 (d, J = 12.0 Hz, 2H), 2.68 (m, 2H), 2.40 (m, 10H), 2.18 (m, 6H), 1.93 (m, 2H), 1.85 (s, 3H), 1.82 (s, 3H), 1.74 (m, 2H).

[0260] Example 9

[0261] Compound 9A:

[0262]

[0263] In a 100 mL single-necked flask, add 2-iodo-5-methylaniline (2.0 g, 8.58 mmol), K3PO4 (2.91 g, 13.73 mmol), Xantphos (0.546 g, 0.944 mmol), Pd(OAc)2 (0.212 g, 0.944 mmol), dimethylphosphite (1.34 g, 17.16 mmol), 36 mL of DMF and 4.8 mL of water. Replace the nitrogen 3 - 4 times, place it at 120 °C and react for about 4 h - 5 h. Detect the reaction by TLC until it is completed. Then cool the reaction solution to room temperature, filter, add 72 mL of H2O to the filtrate, a large amount of solid precipitates, filter, extract the filtrate with 50 mL × 3 dichloromethane, combine the organic phases, wash the organic phases with 50 mL × 2 saturated sodium chloride aqueous solution, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain Compound 9A (1.0 g, yield 64.0%). 11H NMR (400 MHz, CDCl3) δ = 6.96 (dd, J = 13.6, 8.0 Hz, 1H), 6.52 (d, J = 8.0 Hz, 1H), 6.47 (d, J = 4.0 Hz, 1H), 5.29 (s, 2H), 2.26 (s, 3H), 1.76 (s, 3H), 1.73 (s, 3H).

[0264] Compound 9B:

[0265]

[0266] In a 50 mL three-necked flask, compound 9A (500 mg, 2.73 mmol), 5-bromo-2,4-dichloropyrimidine (808 mg, 3.55 mmol), K2CO3 (1.5 g, 10.92 mmol), nBu4NHSO4 (92.69 mg, 0.273 mmol), and 10 mL of DMF were added. The temperature was raised to 65 °C and the reaction was carried out for about 4.5 hours until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 50 mL of H2O was added, and the mixture was extracted with 20 mL of ethyl acetate. The aqueous phase was extracted with 10 mL of ethyl acetate twice. The organic phases were combined, washed successively with 20 mL of water twice and 20 mL of saturated sodium chloride solution twice, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain compound 9B (466 mg, yield 45.7%).

[0267] Compound 9:

[0268]

[0269] Compound 9B (500 mg, 1.33 mmol), compound 1F (383 mg, 1.2 mmol), ethanol solution of 15% hydrogen chloride (974 mg, 4.0 mmol), and 7.5 mL of ethylene glycol monomethyl ether were added to the reaction flask. The reaction was carried out in a sealed tube at 100 °C for 5 h until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 15 mL of saturated aqueous NaHCO3 solution and 10 mL of H2O were added, and a large amount of solid was precipitated. After stirring for another 0.5 h, the mixture was filtered by suction. The filter cake was washed with water and then purified by pulping with 5 mL of a mixed solvent of PE / EA = 2 / 3 to obtain compound 9 (340 mg, yield 43.15%). 11H NMR (400 MHz, CDCl3) δ = 10.38 (s, 1H), 8.25 (d, J = 4.0 Hz, 1H), 8.20 (s, 1H), 7.99 (s, 1H), 7.32 (s, 1H), 7.21 (dd, J = 14.0, 8.0 Hz, 1H), 6.95 (d, J = 7.8 Hz, 1H), 6.62 (s, 1H), 3.86 (s, 3H), 3.13 (d, J = 12.0 Hz, 2H), 2.81 - 2.42 (m, 9H), 2.34 (s, 3H), 2.30 (s, 3H), 2.07 (s, 3H), 1.95 (d, J = 12.0 Hz, 2H), 1.86 (m, 2H), 1.84 (s, 3H), 1.81 (s, 3H), 1.77 - 1.66 (m, 2H). MS-ESI (m / z): 656.2456 (M + H) + 。

[0270] Example 10

[0271] Compound 10A:

[0272]

[0273] In a 250 mL three-necked flask, 2-iodo-3-methylaniline (1.0 g, 4.29 mmol), K3PO4 (1.09 g, 5.15 mmol), Xantphos (0.25 g, 0.429 mmol), Pd(OAc)2 (0.096 g, 0.429 mmol), dimethylphosphite (0.5 g, 6.44 mmol), and 10 mL of DMF were added. The temperature was raised to 120 °C for reaction, and the reaction was monitored by TLC until completion. Subsequently, the reaction solution was cooled to room temperature, filtered by suction. The filtrate was added with 30 mL of H2O and extracted with 30 mL × 3 of dichloromethane. The organic phases were combined, washed once with 50 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain Compound 10A (440.0 mg, yield 56.0%). MS-ESI (m / z): 184.0921 (M + H) + 。

[0274] Compound 10B:

[0275]

[0276] In a 250 mL three-necked flask, add compound 10A (3.93 g, 21.45 mmol), 5-bromo-2,4-dichloropyrimidine (7.33 g, 32.2 mmol), K2CO3 (8.89 g, 64.35 mmol), nBu4NHSO4 (0.73 g, 2.15 mmol), 20 mL of DMF, heat to 65 °C for reaction, and monitor the reaction completion by TLC. After the reaction is cooled to room temperature, add 50 mL of H2O, and a yellow solid precipitates. Continue stirring for about 0.5 h and then filter by suction. Wash the filter cake with 100 mL of H2O, dry to obtain compound 10B (3.88 g, total yield of two steps 48.5%). MS-ESI (m / z): 373.9770 (M + H) + 。

[0277] Compound 10:

[0278]

[0279] Add compound 1F (500 mg, 1.57 mmol), compound 10B (588.12 mg, 1.57 mmol), ethanol solution of 15% hydrogen chloride (917 mg, 3.77 mmol), 7.5 mL of ethylene glycol monomethyl ether to the reaction flask, seal the tube and react at 120 °C, and monitor the reaction completion by TLC. After the reaction solution is cooled to room temperature, add 50 mL of 3% K2CO3 aqueous solution and 30 mL of dichloromethane, stir for 10 minutes, combine the organic phases, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and purify by column chromatography to obtain compound 10 (551 mg, yield 53.5%). MS-ESI (m / z): 656.2486 (M + H) + 。

[0280] Example 11

[0281] Compound 11A:

[0282]

[0283] In a 100 mL three-necked flask, add 2-iodo-4-fluoroaniline (5.0 g, 21.1 mmol), K3PO4 (5.37 g, 25.32 mmol), Xantphos (1.22 g, 2.11 mmol), Pd(OAc)2 (0.474 g, 2.11 mmol), dimethylphosphite (2.47 g, 31.65 mmol), and 50 mL of DMF. Heat the mixture to 120 °C and react for about 4 h until the reaction is completed as detected by TLC. Subsequently, cool the reaction solution to room temperature, filter it by suction. Add 100 mL of H2O to the filtrate, and a large amount of yellow solid precipitates. Filter it by suction again. Extract the filtrate with 50 mL × 4 of dichloromethane. Combine the organic phases. Wash the combined organic phases with 100 mL × 2 of saturated sodium chloride aqueous solution, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain an oily compound 11A. Subsequently, add 4 mL of ethyl acetate and 8 mL of petroleum ether to precipitate solid 11A (1.2 g, yield 30%). 1 1H NMR (400 MHz, CDCl3) δ = 6.98 (m, 1H), 6.78 (m, 1H), 6.61 (m, 1H), 5.23 (s, 2H), 1.79 (s, 3H), 1.75 (s, 3H). MS-ESI (m / z): 188.0639 (M + H) + 。

[0284] Compound 11B:

[0285]

[0286] In a 50 mL three-necked flask, add compound 11A (500 mg, 2.67 mmol), 5-bromo-2,4-dichloropyrimidine (790.94 mg, 3.47 mmol), K2CO3 (1.11 g, 8.01 mmol), nBu4NHSO4 (0.09 g, 0.267 mmol), and 10 mL of DMF. Heat the mixture to 65 °C and react. Monitor the reaction by TLC until it is completed. Subsequently, cool the reaction solution to room temperature, add 50 mL of H2O, extract with 20 mL of ethyl acetate, and extract the aqueous phase with 10 mL of ethyl acetate two more times. Combine the organic phases. Wash the combined organic phases with water twice and with saturated sodium chloride solution twice, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain compound 11B (700 mg, yield 69.3%).

[0287] Compound 11:

[0288]

[0289] Compound 11B (300 mg, 0.79 mmol), compound 1F (303 mg, 0.95 mmol), ethanol solution of 15% hydrogen chloride (578 mg, 2.38 mmol), and 4.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. The reaction was carried out under sealed tube conditions at 120 °C for 5 - 6 h until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 4.5 mL of saturated aqueous NaHCO3 solution and 13.5 mL of H2O were added, and a large amount of solid precipitated. After stirring for an additional 0.5 h, filtration was carried out. The filter cake was washed with water, dried, and purified by column chromatography (DCM / MeOH = 15 / 1 to 10 / 1), and then slurried with 6 mL of a PE / EA = 5 / 1 mixed solvent to obtain compound 11 (90 mg, yield 17.2%). 1 1H NMR (400 MHz, CDCl3) δ = 10.25 (s, 1H), 8.46 (m, 1H), 8.20 (s, 1H), 7.95 (s, 1H), 7.34 (s, 1H), 7.21 (m, 1H), 7.10 - 6.92 (m, 1H), 6.62 (s, 1H), 3.86 (s, 3H), 3.16 (d, J = 11.2 Hz, 2H), 2.84 - 2.47 (m, 9H), 2.37 (m, 5H), 2.16 (s, 3H), 1.97 (d, J = 11.4 Hz, 2H), 1.87 (s, 3H), 1.84 (s, 3H), 1.79 - 1.67 (m, 2H). MS-ESI (m / z): 660.2227 (M + H) + 。

[0290] Example 12

[0291] Compound 12B:

[0292]

[0293] In a 250 mL three-necked flask, 1-bromo-2-fluoro-4-methoxy-5-nitrobenzene (2.0 g, 8.0 mmol), Pd(dppf)Cl2 (0.59 g, 0.8 mmol), triphenylphosphine (0.63 g, 2.4 mmol), cuprous bromide (0.34 g, 2.4 mmol), triethylvinyltin (3.8 g, 12.0 mmol), and 80 mL of toluene were added. The reaction was heated to 110 °C under nitrogen protection and reacted for about 4 h until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 80 mL of saturated potassium fluoride aqueous solution was added to quench the reaction, and 80 mL × 3 of ethyl acetate was used for extraction. The organic phases were combined, washed with 80 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by silica gel column chromatography (petroleum ether / ethyl acetate = 10 / 1) to obtain product 12B (1.41 g, yield 89.2%). 11H NMR (400 MHz, DMSO-d6) δ = 8.24 (d, J = 7.9 Hz, 1H), 7.33 (d, J = 12.6 Hz, 1H), 6.75 (dd, J = 17.7, 11.3 Hz, 1H), 5.95 (d, J = 17.7 Hz, 1H), 5.44 (d, J = 11.3 Hz, 1H), 3.94 (s, 3H).

[0294] Compound 12C:

[0295]

[0296] 12B (1.20 g, 6.09 mmol), 1-methyl-4-(piperidin-4-yl)piperazine (1.34 g, 7.30 mmol), potassium carbonate (1.69 g, 12.17 mmol) and 6 mL of DMF were added to a 50 mL round-bottom flask. The temperature was raised to 120 °C and the reaction was carried out for 4 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature, 20 mL of water was added, and a large amount of yellow solid was precipitated. After stirring for another 1 h, filtration was carried out by suction. The filter cake was washed with water and dried to obtain Compound 12C (1.87 g, yield 85.3%).

[0297] Compound 12A:

[0298]

[0299] Compound 12C (1.8 g, 5.0 mmol) and 0.36 g of 5% Pd / C were weighed into a 150 mL single-necked flask, and 36 mL of methanol was added. The flask was purged with hydrogen 2 - 3 times, and then stirred at room temperature overnight under a hydrogen atmosphere (atmospheric pressure) until the reaction was completed as detected by TLC. Subsequently, after filtration through diatomaceous earth, the filtrate was concentrated under vacuum and purified by column chromatography (DCM / MeOH = 20 / 1) to obtain Compound 12A (780 mg, yield 47.0%). 1 1H NMR (400 MHz, CDCl3) δ = 6.62 (s, 1H), 6.58 (s, 1H), 3.82 (s, 3H), 3.04 - 2.99 (m, 2H), 2.78 - 2.42 (m, 12H), 2.31 (s, 4H), 1.97 - 1.85 (m, 2H), 1.69 - 1.64 (m 2H), 1.16 (t, J = 7.5 Hz, 3H).

[0300] Compound 12:

[0301]

[0302] Compound 2B (565 mg, 1.51 mmol), compound 12A (705 mg, 2.12 mmol), ethanol solution of 15% hydrogen chloride (960 mg, 3.64 mmol), and 7.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 15 mL of saturated aqueous NaHCO3 solution was added, and a large amount of solid was precipitated. After stirring for another 0.5 h, filtration was carried out. The filter cake was washed with water and purified by slurrying to obtain compound 12 (673 mg, yield 66.5%). MS-ESI (m / z): 670.2642 (M + H) + 。 1 H NMR (400 MHz, CDCl3) δ = 10.23 (s, 1H), 8.25 (dd, J = 8.4, 4.8 Hz, 1H), 8.19 (s, 1H), 8.06 (s, 1H), 7.44 - 7.29 (m, 2H), 7.11 (d, J = 14.0 Hz, 1H), 6.66 (s, 1H), 3.85 (s, 3H), 3.08 (d, J = 11.6 Hz, 2H), 2.93 - 2.21 (m, 19H), 1.95 (d, J = 12.0 Hz, 2H), 1.82 (d, J = 13.2 Hz, 6H), 1.78 - 1.66 (m, 2H), 1.00 (t, J = 7.6 Hz, 3H).

[0303] Example 13

[0304] Compound 13A:

[0305]

[0306] Referring to the synthesis method of compound 12A, only the triethylvinyltin in it was replaced with 2-(tributylstannyl)propene to obtain compound 13A.

[0307] Compound 13:

[0308]

[0309] Compound 2B (565 mg, 1.51 mmol), compound 13A (735 mg, 2.12 mmol), ethanol solution of 15% hydrogen chloride (960 mg, 3.64 mmol), and 7.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 15 mL of saturated aqueous NaHCO3 solution was added, and a large amount of solid precipitated. After stirring for an additional 0.5 h, the mixture was filtered by suction. The filter cake was washed with water and purified by slurrying to obtain compound 13 (622 mg, yield 60.2%). MS-ESI (m / z): 684.2756 (M + H) + 。

[0310] Example 14

[0311] Compound 14A:

[0312]

[0313] 1-Chloro-2-fluoro-4-methoxy-5-nitrobenzene (500 mg, 2.43 mmol), 1-methyl-4-(4-piperidinyl)piperazine (490 mg, 2.68 mmol), potassium carbonate (507 mg, 3.65 mmol), and 10 mL of DMF were added to a 100 mL round-bottom flask. The temperature was raised to 80 °C and the reaction was carried out for 2.5 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 30 mL of water was added, and a large amount of yellow solid precipitated. After stirring for 1 h, the mixture was filtered by suction. The filter cake was washed with water and dried to obtain compound 14A (740 mg, yield 82.5%).

[0314] Compound 14B:

[0315]

[0316] Compound 14A (740 mg, 2.01 mmol), reduced iron powder (672 mg, 12.04 mmol), ammonium chloride (75 mg, 1.40 mmol), and 67 mL of ethanol / water = 3 / 1 mixed solution were added to a 250 mL round-bottom flask. The temperature was raised to reflux and the reaction was carried out for about 6.5 h until the reaction was completed as detected by TLC. The reaction solution was cooled to room temperature, and the pH of the reaction solution was adjusted to about 8 with 2 mol / L aqueous sodium hydroxide solution. After concentration to dryness, 110 mL of dichloromethane / methanol = 10 / 1 mixed solution was added and stirred at room temperature for about 0.5 h. Then, it was filtered by suction. The filtrate was concentrated and separated by column chromatography (dichloromethane / methanol = 8 / 1) to obtain compound 14B (647 mg, yield 95.2%). MS-ESI (m / z): 339.1928 (M + H) + 。

[0317] Compound 14:

[0318]

[0319] Compound 2B (253 mg, 0.67 mmol), compound 14B (243 mg, 1.01 mmol), ethanol solution of 15% hydrogen chloride (493 mg, 2.02 mmol), and 4 mL of ethylene glycol monomethyl ether were added to a reaction flask. The reaction was carried out in a sealed tube at 120 °C for 5 - 6 h until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 4 mL of saturated aqueous NaHCO3 solution and 12 mL of H2O were added, and a large amount of solid precipitated. After stirring for an additional 0.5 h, filtration was performed. The filter cake was washed with water, dried, and purified by column chromatography (DCM / MeOH = 15 / 1 to 10 / 1), and then slurried with 6 mL of a mixed solvent of PE / EA = 5 / 1 to obtain compound 14 (120 mg, yield 26.3%). 1 1H NMR (400 MHz, CDCl3) δ = 10.38 (s, 1H), 8.39 - 8.24 (m, 2H), 8.19 (s, 1H), 7.47 (d, J = 8.8 Hz, 1H), 7.41 (s, 1H), 7.10 (d, J = 14.4 Hz, 1H), 6.61 (s, 1H), 3.88 (s, 3H), 3.40 (d, J = 11.2 Hz, 2H), 2.83 - 2.46 (m, 9H), 2.38 (s, 3H), 2.33 (s, 3H), 2.16 (s, 2H), 1.95 (d, J = 11.4 Hz, 2H), 1.81 (m, 8H). MS-ESI (m / z): 676.1902 (M + H) + 。

[0320] Example 15

[0321] Compound 15A:

[0322]

[0323] Add 2-chloro-4-fluoro-5-nitrotoluene (10.0 g, 52.4 mmol), cesium carbonate (83.2 g, 262 mmol), and 100 mL of isopropanol into a 250 mL single-necked flask. Place it in a reaction at 60 °C for 4 h, and monitor by TLC until the raw materials disappear. Subsequently, cool the reaction solution to room temperature, add 100 mL of ethyl acetate, stir for 10 min and then filter. Wash the filter cake with 50 mL of ethyl acetate, concentrate it under reduced pressure to dryness. Add 100 mL of water and 150 mL of ethyl acetate to the residue, let it stand for phase separation. Extract the aqueous phase with 100 mL of ethyl acetate once, combine the ethyl acetate phases. Then wash the ethyl acetate phase successively with 100 mL×2 of water and 100 mL×2 of saturated sodium chloride aqueous solution, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain compound 15A (11.45 g, yield 95.4%).

[0324] Compound 15B:

[0325]

[0326] Add compound 15A (9.0 g, 39.2 mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (13.33 g, 43.12 mmol), K2CO3 (16.25 g, 117.6 mmol), 82 mL of DME, and 8 mL of water into a 250 mL single-necked flask. Replace the air with nitrogen three times, then add Pd(dppf)Cl2 (2.86 g, 3.92 mmol), and replace the air with nitrogen three times again. Subsequently, react at 120 °C for 5 h, and monitor by TLC until the raw materials disappear. Cool the reaction solution to room temperature, filter, wash the filter cake with 50 mL of ethyl acetate, concentrate the filtrate under reduced pressure to dryness. Purify the residue by column chromatography (PE∶EA = 20∶1 - 10∶1) to obtain a pale yellow oily compound 15B (8.8 g, yield 60.0%). 1H NMR (400 MHz, CDCl3) δ = 7.62 (s, 1H), 6.79 (s, 1H), 5.62 (s, 1H), 4.63 (m, 1H), 4.06 (d, J = 2.0 Hz, 2H), 3.64 (t, J = 5.6 Hz, 2H), 2.33 (s, 2H), 2.24 (s, 3H), 1.51 (s, 9H), 1.37 (d, J = 6.0 Hz, 6H). MS-ESI (m / z): 399.1882 (M+Na) + 。

[0327] Compound 15C:

[0328]

[0329] Add compound 15B (8.8 g, 23.4 mmol) and 20 mL of ethyl acetate to a 150-mL single-necked flask, stir to dissolve, then add 25 mL of ethyl acetate solution of hydrogen chloride (15%), and stir at room temperature overnight. TLC detection shows that the raw materials disappear. Add 100 mL of water to the reaction solution, stir for 10 min, let it stand for liquid separation, wash the ethyl acetate layer with 40 mL of water again, combine the aqueous phases, then adjust the pH to 8 with saturated sodium bicarbonate solution, extract with 100 mL of ethyl acetate, extract the aqueous phase with 50 mL of ethyl acetate twice more, combine the ethyl acetate phases, then wash the ethyl acetate phase with saturated sodium chloride solution twice, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain compound 15C (4.2 g, yield 65.01%). 1 1H NMR (400 MHz, CDCl3) δ = 7.63 (s, 1H), 6.84 (s, 1H), 5.68 (s, 1H), 4.64 (m, 1H), 3.89 (d, J = 2.4 Hz, 2H), 3.48 (t, J = 6.0 Hz, 2H), 2.71 (m, 2H), 2.29 (s, 3H), 1.38 (d, J = 6.0 Hz, 6H). MS-ESI (m / z): 277.1547 (M+H) + 。

[0330] Compound 15D:

[0331]

[0332] Add compound 15C (1.60 g, 5.79 mmol) and 64 mL of dichloromethane to a 150-mL single-necked flask to dissolve, then add tetrahydro-4H-pyran-4-one (1.65 g, 17.37 mmol), acetic acid (1.32 g, 23.16 mmol), and sodium triacetoxyborohydride (4.66 g, 23.16 mmol), and stir at room temperature overnight. TLC detection shows that the raw materials disappear. Subsequently, add 120 mL of water to the reaction solution, stir for 10 min, let it stand for liquid separation, extract the aqueous phase with 20 mL of dichloromethane, combine the two dichloromethane phases, then wash the dichloromethane phase successively with 40 mL of water and 40 mL of saturated sodium chloride solution, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain oily compound 15D (2.00 g, yield 95.80%).

[0333] Compound 15E:

[0334]

[0335] Add compound 15D (0.99 g, 2.77 mmol) into a 50 mL single-necked flask, dissolve it with 20 mL of ethanol, then add 2.5 mL of hydrochloric acid solution (2N) and iron powder (0.994 g, 17.76 mmol), and react at 60 °C for 3 h. Monitor the reaction by TLC until the raw materials disappear. After the reaction is completed, cool the reaction solution to room temperature, filter it, concentrate the filtrate under reduced pressure, and purify it by column chromatography to obtain an oily substance 15E (779 mg, yield 85.0%). MS-ESI (m / z): 331.2381 (M+H) + 。

[0336] Compound 15:

[0337]

[0338] Add compound 2B (315 mg, 0.84 mmol), compound 15E (417 mg, 1.26 mmol), ethanol solution of 15% hydrochloric acid (613 mg, 2.52 mmol), and 5.0 mL of ethylene glycol monomethyl ether into a reaction flask, then seal the tube and react at 120 °C for 5 - 6 h. After monitoring the reaction by TLC until it is completed, cool the reaction solution to room temperature, then add 50 mL of 3% K2CO3 aqueous solution and 50 mL of dichloromethane, stir for 10 minutes, and then let it stand for liquid separation. The upper aqueous phase is washed with 50 mL of dichloromethane again. Combine the two dichloromethane phases, then wash the dichloromethane phase with 50 mL of saturated brine, dry it over anhydrous sodium sulfate, filter it, concentrate the filtrate, and purify it by column chromatography to obtain compound 15 (310 mg, yield 55.20%). MS-ESI (m / z): 668.2334 (M+H) + 。

[0339] Example 16

[0340] Compound 16D:

[0341]

[0342] Referring to the synthesis method of reference compound 15D, 4.20 g (15.20 mmol) of compound 15C was added into a 250 mL single-necked flask and dissolved in 160 mL of dichloromethane. Then, N-methyl-4-piperidone (5.16 g, 45.60 mmol), acetic acid (3.66 g, 60.80 mmol), and sodium triacetoxyborohydride (12.88 g, 60.80 mmol) were added, and the mixture was stirred overnight at room temperature. TLC detection showed that the raw materials disappeared. Subsequently, 60 mL of water was added to the reaction solution, stirred for 10 min, allowed to stand for phase separation, and the aqueous phase was extracted with 60 mL of dichloromethane again. The two dichloromethane phases were combined, and then the dichloromethane phase was washed successively with 40 mL of water and 40 mL of saturated sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain an oily compound 16D (4.51 g, yield 79.4%). MS-ESI (m / z): 374.2443 (M+H) + 。

[0343] Compound 16E:

[0344]

[0345] Referring to the synthesis method of reference compound 15E, 1.0 g (2.68 mmol) of compound 16D was added into a 50 mL single-necked flask and dissolved in 20 mL of ethanol. Then, 2.5 mL (2N) of hydrochloric acid solution and iron powder (0.960 g, 17.15 mmol) were added, and the reaction was carried out at 60 °C for 3 h. TLC detection showed that the raw materials disappeared. After the reaction was completed, the reaction solution was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure and purified by column chromatography to obtain an oily substance 16E (736 mg, yield 80.0%).

[0346] Compound 16:

[0347]

[0348] Compound 2B (400 mg, 1.07 mmol), compound 16E (333 mg, 0.97 mmol), ethanol solution of 15% hydrogen chloride (708 mg, 2.91 mmol), and 6 mL of ethylene glycol monomethyl ether were added to the reaction flask, and the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h until TLC detection showed that the reaction was completed. Subsequently, the reaction solution was cooled to room temperature, 6 mL of saturated NaHCO3 aqueous solution and 6 mL of H2O were added, and the mixture was extracted with 30 mL×3 of dichloromethane. After the organic phases were combined, they were washed with 30 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography separation (DCM / MeOH = 10 / 1) to obtain compound 16 (120 mg, yield 16.5%). MS-ESI (m / z): 681.2644 (M+H) + 。

[0349] Example 17

[0350] Compound 17A:

[0351]

[0352] In a 100 mL single-necked flask, add compound 15D (1.80 g, 5.00 mmol), 0.27 g of 5% Pd / C, and 30 mL of methanol. Replace the gas with hydrogen 2 - 3 times, stir at room temperature overnight under a hydrogen atmosphere until the reaction is completed as detected by TLC. Then, filter through diatomaceous earth, concentrate the filtrate under vacuum, and purify by column chromatography to obtain compound 17A (1.51 g, yield 90.8%). MS-ESI (m / z): 333.2572 (M + H) + 。

[0353] Compound 17:

[0354]

[0355] Add compound 2B (566 mg, 1.51 mmol), compound 17A (706 mg, 2.12 mmol), ethanol solution of 15% hydrogen chloride (960 mg, 3.64 mmol), and 4.5 mL of ethylene glycol monomethyl ether to the reaction flask. Then, seal the tube and react at 120 °C for 5 - 6 h. After the reaction is completed as detected by TLC, cool the reaction solution to room temperature, then add 15 mL of saturated aqueous NaHCO3 solution to precipitate a solid. Continue to stir for 0.5 h and then filter by suction. Wash the filter cake with water, and purify the filter cake by slurrying with 20 mL of a mixed solvent of EA / PET = 1 / 5 to obtain compound 17 (558 mg, yield 55.2%). MS-ESI (m / z): 670.2554 (M + H) + 。

[0356] Example 18

[0357] Compound 18A:

[0358]

[0359] In a 100 mL single-necked flask, add compound 16D (1.60 g, 4.28 mmol), 1.0 g of 5% Pd / C, 10 mL of methanol, replace the gas with hydrogen 2 - 3 times, stir at room temperature for 48 h under a hydrogen atmosphere, and filter through diatomaceous earth after the reaction is completed as detected by TLC. Then, concentrate the filtrate under vacuum and purify by column chromatography to obtain compound 18A (1.3 g, yield 87.8%).

[0360] Compound 18:

[0361]

[0362] Compound 2B (570 mg, 1.52 mmol), compound 18A (500 mg, 1.45 mmol), ethanol solution of 15% hydrogen chloride (1.06 g, 4.35 mmol), and 7.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 120 °C for 4 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 15 mL of saturated aqueous NaHCO3 was added to precipitate a solid. Stirring was continued for 0.5 h, and then filtration was carried out. The filter cake was dissolved in methanol, concentrated under reduced pressure to obtain an oil. After dissolving the oil in 3 mL of ethyl acetate, 2 mL of petroleum ether was added dropwise and the mixture was slurried for purification to obtain compound 18 (241 mg, yield 23.21%). 1 1H NMR (400 MHz, CDCl3) δ = 10.30 (s, 1H), 8.35 (dd, J = 8.4, 4.4 Hz, 1H), 8.19 (s, 1H), 8.05 (s, 1H), 7.51 (s, 1H), 7.33 (d, J = 8.4 Hz, 1H), 7.12 (d, J = 14.0 Hz, 1H), 6.82 (s, 1H), 4.52 (m, 1H), 3.06 (d, J = 11.2 Hz, 2H), 2.96 (d, J = 11.2 Hz, 2H), 2.71 - 2.60 (m, 1H), 2.38 (s, 3H), 2.34 (m, 2H), 2.29 (s, 3H), 2.16 (s, 3H), 2.01 - 1.91 (m, 4H), 1.84 (m, 8H), 1.72 (m, 5H), 1.37 (s, 3H), 1.35 (s, 3H). MS-ESI (m / z): 683.2800 (M+H) + 。

[0363] Example 19

[0364] Compound 19B:

[0365]

[0366] Add compound 2-nitro-4-methyl-5-bromoanisole (9.65 g, 39.2 mmol), N-Boc-1,2,5,6-tetrahydropyridine-4-boronic acid pinacol ester (13.33 g, 43.12 mmol), K2CO3 (16.25 g, 117.6 mmol), DME 82 ml, and water 8 ml into a 250 mL single-necked flask. Replace the air with nitrogen three times, then add Pd(dppf)Cl2 (2.86 g, 3.92 mmol), and replace the air with nitrogen three more times. Subsequently, react at 120 °C for 5 h and monitor the reaction by TLC until the starting materials disappear. Cool the reaction mixture to room temperature, filter, wash the filter cake with 50 ml of ethyl acetate, concentrate the filtrate under reduced pressure to dryness, and purify the residue by column chromatography to obtain a pale yellow oily compound 19B (7.65 g, yield 56.0%). MS-ESI (m / z): 349.1801 (M+H) + 。

[0367] Compound 19C:

[0368]

[0369] Refer to the synthesis method of compound 15C, and simply replace compound 15B therein with compound 19B to obtain compound 19C.

[0370] Compound 19D:

[0371]

[0372] Refer to the synthesis method of compound 15D, and simply replace compound 15C therein with compound 19C to obtain compound 19D. MS-ESI (m / z): 333.1875 (M+H) + 。

[0373] Compound 19E:

[0374]

[0375] Refer to the synthesis method of compound 17A, and simply replace compound 15D therein with compound 19D to obtain compound 19E. MS-ESI (m / z): 305.2285 (M+H) + 。

[0376] Compound 19:

[0377]

[0378] Refer to the synthesis method of compound 17, and simply replace compound 17A therein with compound 19E to obtain compound 19. MS-ESI (m / z): 642.2173 (M+H) +。

[0379] Example 20

[0380] Compound 20D:

[0381]

[0382] Referring to the synthesis method of reference compound 16D, only replace compound 15C therein with compound 19C to obtain compound 20D.

[0383] Compound 20E:

[0384]

[0385] Referring to the synthesis method of reference compound 17A, only replace compound 15D therein with compound 20D to obtain compound 20E. MS-ESI (m / z): 318.2512 (M+H) + 。

[0386] Compound 20:

[0387]

[0388] Referring to the synthesis method of reference compound 17, only replace compound 17A therein with compound 20E to obtain compound 20. MS-ESI (m / z): 655.2553 (M+H) + 。

[0389] Example 21

[0390] Compound 21A:

[0391]

[0392] Add compound 15C (4.20 g, 15.20 mmol) into a 250 mL single-necked flask, dissolve it with 160 ml of dichloromethane, then add paraformaldehyde (1.37 g, 45.60 mmol), acetic acid (3.66 g, 60.80 mmol), and sodium triacetoxyborohydride (12.88 g, 60.80 mmol), and stir overnight at room temperature. TLC detection shows that the raw materials disappear. Subsequently, add 60 ml of water to the reaction solution, stir for 10 min, let it stand for phase separation, extract the aqueous phase with 60 ml of dichloromethane again, combine the two dichloromethane phases, and then wash the dichloromethane phase successively with 40 ml of water and 40 ml of saturated sodium chloride solution, dry it over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain oily compound 21A (3.58 g, yield 81.2%).

[0393] Compound 21B:

[0394]

[0395] Referring to the synthesis method of reference compound 17A, only replace compound 15D therein with compound 21A to obtain compound 21B. MS-ESI (m / z): 263.2182 (M+H) + 。

[0396] Compound 21:

[0397]

[0398] Referring to the synthesis method of reference compound 17, only replace compound 17A therein with compound 21B to obtain compound 21. MS-ESI (m / z): 600.2165 (M+H) + 。

[0399] Example 22

[0400] Compound 22A:

[0401]

[0402] In a 100 mL single-necked flask, add compound 15C (1.38 g, 5.00 mmol), 0.21 g of 5% Pd / C, and 30 mL of methanol. Replace with hydrogen 2-3 times, stir at room temperature overnight under a hydrogen atmosphere until the reaction is complete as detected by TLC. Then, filter through diatomaceous earth, concentrate the filtrate under vacuum, and purify by column chromatography to obtain compound 22A (1.07 g, yield 85.8%). MS-ESI (m / z): 249.1987 (M+H) + 。

[0403] Compound 22:

[0404]

[0405] Referring to the synthesis method of reference compound 17, only replace compound 17A therein with compound 22A to obtain compound 22. MS-ESI (m / z): 586.1975 (M+H) + 。

[0406] Example 23

[0407] Compound 23A:

[0408]

[0409] In a 100 mL three-necked flask, add 5-fluoro-4-methyl-2-nitrophenol (2.57 g, 15.00 mmol), 60% sodium hydride (2.40 g, 60.00 mmol), and 30 mL of DMF. After stirring at 0 °C for 30 minutes, add 2-iodo-1,1,1-trifluoroethane (4.73, 22.50 mmol) in portions. Then, restore to room temperature and stir overnight. Monitor the reaction by TLC until completion. Subsequently, add 60 mL of water and 100 mL of ethyl acetate to the reaction mixture, stir for 10 minutes, and let it stand for liquid separation. Wash the aqueous phase with 50 mL of ethyl acetate once more. Combine the two ethyl acetate phases. Then, wash the organic phase with saturated brine once, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and obtain compound 23A (2.68 g, yield 70.5%) after column chromatography.

[0410] Compound 23B:

[0411]

[0412] In a 250 mL three-necked flask, add compound 23A (2.53 g, 10.00 mmol), K2CO3 (4.14 g, 30.00 mmol), 1-methyl-4-(4-piperidinyl)piperazine (2.75 g, 15.00 mmol), and 30 mL of DMF. Heat to 120 °C and stir the reaction for about 4 hours until the reaction is complete as monitored by TLC. Then, cool the reaction mixture to room temperature, add 60 mL of water and 100 mL of ethyl acetate, stir for 10 minutes, and let it stand for liquid separation. Wash the aqueous phase with 50 mL of ethyl acetate once more. Combine the two ethyl acetate phases. Then, wash the organic phase with saturated brine once, dry over anhydrous sodium sulfate, filter, concentrate the filtrate, and obtain compound 23B (2.47 g, yield 59.2%). MS-ESI (m / z): 417.2175 (M+H) + 。

[0413] Compound 23C:

[0414]

[0415] In a 100 mL single-necked flask, add compound 23B (2.47 g, 5.92 mmol), 0.25 g of 5% Pd / C, and 30 mL of methanol. Replace the air with hydrogen 2 - 3 times, and stir at room temperature overnight in a hydrogen atmosphere until the reaction is complete as monitored by TLC. Then, filter through diatomaceous earth, concentrate the filtrate under vacuum, and purify by column chromatography to obtain compound 23C (1.93 g, yield 84.5%).

[0416] Compound 23:

[0417]

[0418] Referring to the synthesis method of reference compound 17, only compound 17A therein was replaced with compound 23C to obtain compound 23. MS-ESI (m / z): 724.2315 (M+H) + 。 1 H NMR (400 MHz, CDCl3) δ = 10.56 (s, 1H), 8.42 (dd, J = 8.8, 4.8 Hz, 1H), 8.23 (s, 1H), 8.10 (s, 1H), 7.42 (m, 2H), 7.16 (d, J = 14.4 Hz, 1H), 6.82 (s, 1H), 4.85 (m, 2H), 3.10 (d, J = 11.6 Hz, 2H), 2.80 - 2.48 (m, 11H), 2.36 (s, 3H), 2.32 (s, 3H), 2.15 (s, 3H), 2.05 (d, J = 11.0 Hz, 2H), 1.88 (s, 3H), 1.84 (s, 3H), 1.78 - 1.64 (m, 2H).

[0419] Example 24

[0420] Compound 24D:

[0421]

[0422] Into a 50 mL single-necked flask, add compound 1C (2.00 g, 11.69 mmol), cesium carbonate (5.74 g, 17.52 mmol), 14 mL of DMF, and 0.34 mL of H2O. After stirring evenly at room temperature, add sodium difluorochloroacetate (3.56 g, 23.38 mmol) in portions. After addition, heat the reaction mixture to 100 °C and stir for about 2 hours until the reaction is complete as detected by TLC. Subsequently, cool the reaction mixture to room temperature, add 56 mL of H2O, extract with 28 mL × 2 of petroleum ether, combine the organic phases, wash with 20 mL of saturated brine, dry over anhydrous sodium sulfate, filter, and concentrate the filtrate to obtain compound 24D (1.49 g, yield 57.64%).

[0423] Compound 24E:

[0424]

[0425] In a 50 mL single-necked flask, add compound 24D (1.21 g, 5.45 mmol), add 22.35 mL of DMF and stir to dissolve. Subsequently, add K2CO3 (1.51 g, 10.91 mmol) and 1-methyl-4(4-piperidyl)-piperazine (1.30 g, 7.09 mmol). Heat the temperature to 120 °C and react for about 2 h until the reaction is completed by TLC detection. Subsequently, cool the reaction solution to room temperature, add 40 mL of water, and extract with 120 mL × 2 of EA. Combine the organic phases. The organic phases are washed successively with 30 mL × 3 of H2O and 30 mL of saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, and the filtrate is concentrated to obtain compound 24E (1.91 g, yield 91.3%).

[0426] Compound 24F:

[0427]

[0428] In a 100 mL single-necked flask, add compound 24E (1.91 g, 4.97 mmol), 0.24 g of 5% Pd / C, 48 mL of methanol, and replace with hydrogen 2 - 3 times. Stir at room temperature overnight under a hydrogen atmosphere (atmospheric pressure) until the reaction is completed by TLC detection. After filtration through diatomaceous earth, the filtrate is concentrated in vacuo and purified by column chromatography to obtain compound 24F (1.68 g, yield 95.5%). MS-ESI (m / z): 355.2302 (M + H) + 。

[0429] Compound 24:

[0430]

[0431] Referring to the synthesis method of compound 2, only replace compound 1F therein with compound 24F to obtain compound 24. MS-ESI (m / z): 692.2234 (M + H) + 。 1 1H NMR (400 MHz, CDCl3) δ = 10.50 (s, 1H), 8.42 (m, 1H), 8.18 (s, 1H), 8.06 (s, 1H), 7.50 - 7.36 (m, 2H), 7.12 (d, J = 14.0 Hz, 1H), 6.80 (s, 1H), 6.66 (s, 0.25H), 6.52 (s, 0.5H), 6.40 (s, 0.25H), 3.16 (d, J = 11.6 Hz, 2H), 2.82 - 2.42 (m, 11H), 2.36 (s, 3H), 2.34 (s, 3H), 2.16 (s, 3H), 1.96 (d, J = 11.0 Hz, 2H), 2.12 (s, 3H), 1.96 (s, 3H), 1.78 - 1.66 (m, 2H).

[0432] Example 25

[0433] Compound 25A:

[0434]

[0435] Compound 1B (2.13 g, 6.46 mmol), 4-fluoro-2-methoxy-5-nitroaniline (1.44 g, 7.75 mmol), ethanol solution of 15% hydrogen chloride (4.71 g, 19.37 mmol), and 30 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the temperature was raised to 120 °C and the reaction was carried out under sealed tube conditions for about 5 hours until the reaction was completed as detected by TLC. The reaction solution was cooled to room temperature, 90 mL of about 5% aqueous sodium bicarbonate solution was added, and a large amount of brownish-yellow solid was precipitated. After stirring for about 0.5 hours, filtration was carried out, washed with water, and the filter cake was purified by slurrying to obtain Compound 25A (2.09 g, yield 67.4%).

[0436] Compound 25B:

[0437]

[0438] In a 150 mL three-necked flask, Compound 25A (1.57 g, 3.27 mmol), K2CO3 (0.91 g, 6.53 mmol), 1-methyl-4-(4-piperidyl)piperazine (0.72 g, 3.92 mmol), and 30 mL of DMF were added. The temperature was raised to 120 °C and stirred for about 3 hours until the reaction was completed as detected by TLC. The reaction solution was cooled to room temperature, 60 mL of H2O was added, and a large amount of solid was precipitated. After stirring for about 0.5 hours, filtration was carried out, washed with water, and the filter cake was dried to obtain Compound 25B (1.69 g, yield 80.5%). MS-ESI (m / z): 643.2758 (M+H) + 。

[0439] Compound 25C:

[0440]

[0441] In a 100 mL single-necked flask, add compound 25B (1.03 g, 1.61 mmol), reduced iron powder (0.54 g, 9.64 mmol), and ammonium chloride (0.06 g, 1.12 mmol). After adding 90 mL of a mixed solvent of EtOH / H2O = 3 / 1 and stirring evenly, reflux and react for about 2 h until the reaction is completed as detected by TLC. Cool the reaction solution to room temperature. After concentrating the reaction solution, add 330 mL of a mixed solvent of DCM / MeOH = 10 / 1 and stir for about 15 minutes, then filter. The filtrate is concentrated to obtain compound 25C (0.88 g, yield 89.7%). MS-ESI (m / z): 613.2968 (M+H) + 。

[0442] Compound 25:

[0443]

[0444] In a 50 mL single-necked flask, add compound 25C (514 mg, 0.84 mmol) and DIPEA (120 mg, 0.93 mmol), and dissolve them with 7.5 mL of dichloromethane and cool to 0 - 10 °C; weigh acryloyl chloride (92 mg, 1.01 mmol), dilute it with 2.5 mL of DCM, and slowly add it dropwise to the above reaction solution. After the addition is complete, continue to react at 0 - 10 °C for about 3 h until the reaction is completed as detected by TLC. Subsequently, add 20 mL of dichloromethane and 20 mL of H2O to the reaction solution, separate the layers, wash the dichloromethane layer with 20 mL of saturated sodium chloride aqueous solution, dry it over anhydrous sodium sulfate, filter, and concentrate the filtrate, then purify it by column chromatography to obtain compound 25 (189 mg, yield 33.9%). MS-ESI (m / z): 667.3085 (M+H) + 。

[0445] Example 26

[0446] Compound 26B:

[0447]

[0448] Referring to the synthesis method of compound 25B, only replace compound 1-methyl-4-(4-piperidinyl)piperazine with compound 4-dimethylaminopiperidine to obtain compound 26B. MS-ESI (m / z): 588.2283 (M+H) + 。

[0449] Compound 26C:

[0450]

[0451] Referring to the synthesis method of reference compound 25C, compound 26C was obtained. MS-ESI (m / z): 558.2412 (M+H) + .

[0452] Compound 26:

[0453]

[0454] Referring to the synthesis method of reference compound 25, only replacing compound 25C therein with compound 26C, compound 26 was obtained. MS-ESI (m / z): 612.2684 (M+H) + .

[0455] Example 27

[0456] Compound 27B:

[0457]

[0458] Referring to the synthesis method of reference compound 25B, only replacing 1-methyl-4-(4-piperidinyl)piperazine with compound N,N,N'-trimethylethylenediamine, compound 27B was obtained. MS-ESI (m / z): 562.2127 (M+H) + .

[0459] Compound 27C:

[0460]

[0461] Referring to the synthesis method of reference compound 25C, compound 27C was obtained. MS-ESI (m / z): 532.2322 (M+H) + .

[0462] Compound 27:

[0463]

[0464] Referring to the synthesis method of reference compound 25, only replacing compound 25C therein with compound 27C, compound 27 was obtained. MS-ESI (m / z): 586.2465 (M+H) + .

[0465] Example 28

[0466] Compound 28A:

[0467]

[0468] In a 250 mL three-necked flask, weigh in compound 1D (4.5 g, 24.3 mmol), K2CO3 (6.71 g, 48.6 mmol), morpholine (3.17 g, 36.4 mmol), 67.5 mL of DMF. Heat up to 120 °C and stir the reaction for about 4 hours until the reaction is completed as detected by TLC. Then cool to room temperature, add 130 mL of H2O to the reaction solution, a large amount of solid precipitates, continue to stir for about 0.5 hours and then filter by suction. Wash the filter cake with water and dry to obtain compound 28A (3.26 g, yield 53.1%). MS-ESI (m / z): 253.1148 (M+H) + 。

[0469] Compound 28B:

[0470]

[0471] In a 250 mL single-necked flask, weigh in compound 28A (3.26 g, 12.94 mmol), 0.49 g of 5% Pd / C, 60 mL of methanol, displace with hydrogen 2 - 3 times, stir at room temperature overnight under a hydrogen atmosphere (atmospheric pressure) until the reaction is completed as detected by TLC. Then filter through diatomaceous earth, concentrate the filtrate under vacuum, and purify by column chromatography to obtain compound 28B (2.67 g, yield 92.8%). MS-ESI (m / z): 223.1475 (M+H) + 。

[0472] Compound 28:

[0473]

[0474] Add compound 2B (566 mg, 1.51 mmol), compound 28B (471 mg, 2.12 mmol), ethanol solution of 15% hydrogen chloride (960 mg, 3.64 mmol), and 4.5 mL of ethylene glycol monomethyl ether to the reaction flask. Then seal the tube and react at 120 °C for 5 - 6 h. After the reaction is completed as detected by TLC, cool the reaction solution to room temperature, then add 15 mL of saturated NaHCO3 aqueous solution, a large amount of solid precipitates, continue to stir for 0.5 hours and then filter by suction. Wash the filter cake with water, and purify the filter cake by slurrying with 20 mL of EA / PET = 1 / 5 mixed solvent to obtain compound 28 (387 mg, yield 45.7%). MS-ESI (m / z): 560.1402 (M+H) + 。

[0475] Example 29

[0476] Compound 29A:

[0477]

[0478] In a 250 mL three-necked flask, weigh in compound 1D (4.5 g, 24.3 mmol), K2CO3 (6.71 g, 48.6 mmol), N-methylpiperazine (3.64 g, 36.4 mmol), and 67.5 mL of DMF. Heat up to 120 °C and stir the reaction for about 4 hours until the reaction is completed as detected by TLC. Then cool to room temperature, add 130 mL of H2O to the reaction solution, a large amount of solid precipitates. Continue to stir for about 0.5 hours and then filter by suction. Wash the filter cake with water and dry to obtain compound 29A (3.38 g, yield 52.5%). MS-ESI (m / z): 266.1547 (M+H) + 。

[0479] Compound 29B:

[0480]

[0481] In a 250 mL single-necked flask, weigh in compound 29A (3.38 g, 12.71 mmol), 0.51 g of 5% Pd / C, and 60 mL of methanol. Replace the gas with hydrogen 2 - 3 times, and stir at room temperature overnight under a hydrogen atmosphere (atmospheric pressure) until the reaction is completed as detected by TLC. Then filter through diatomaceous earth, concentrate the filtrate under vacuum, and purify by column chromatography to obtain compound 29B (2.80 g, yield 93.4%). MS-ESI (m / z): 236.1791 (M+H) + 。

[0482] Compound 29:

[0483]

[0484] Add compound 2B (566 mg, 1.51 mmol), compound 29B (500 mg, 2.12 mmol), 15% hydrochloric acid in ethanol solution (960 mg, 3.64 mmol), and 4.5 mL of ethylene glycol monomethyl ether to the reaction flask. Then seal the tube and react at 120 °C for 5 - 6 h. After the reaction is completed as detected by TLC, cool the reaction solution to room temperature. Then add 15 mL of saturated NaHCO3 aqueous solution, a large amount of solid precipitates. Continue to stir for 0.5 hours and then filter by suction. Wash the filter cake with water, and purify the filter cake by slurrying with 20 mL of EA / PET = 1 / 5 mixed solvent to obtain compound 29 (350 mg, yield 40.5%). MS-ESI (m / z): 573.1728 (M+H) + 。 11H NMR (400 MHz, CDCl3) δ = 10.31 (s, 1H), 8.34 (dd, J = 8.6, 4.6 Hz, 1H), 8.19 (s, 1H), 8.05 (s, 1H), 7.39 - 7.32 (m, 2H), 7.15 - 7.08 (m, 1H), 6.67 (s, 1H), 3.87 (s, 3H), 2.95 (t, J = 4.8 Hz, 4H), 2.63 (s, 4H), 2.40 (d, J = 5.0 Hz, 6H), 2.17 (s, 3H), 1.84 (d, J = 13.1 Hz, 6H). MS-ESI (m / z): 573.1728 (M+H) + 。

[0485] Example 30

[0486] Compound 30A:

[0487]

[0488] In a 250 mL three-necked flask, weigh in compound 1D (4.5 g, 24.3 mmol), K2CO3 (6.71 g, 48.6 mmol), 4-(4-piperidyl)morpholine (6.20 g, 36.4 mmol), and 67.5 mL of DMF. Heat to 120 °C and stir the reaction for about 4 hours until the reaction is complete as detected by TLC. Then cool to room temperature, add 130 mL of H2O to the reaction solution, a large amount of solid precipitates, continue to stir for about 0.5 hours and then filter by suction. Wash the filter cake with water and dry to obtain compound 30A (5.54 g, yield 67.8%). MS-ESI (m / z): 336.1944 (M+H) + 。

[0489] Compound 30B:

[0490]

[0491] In a 250 mL single-necked flask, weigh in compound 30A (5.54 g, 16.48 mmol), 0.83 g of 5% Pd / C, and 60 mL of methanol. Replace the gas with hydrogen 2 - 3 times, stir at room temperature overnight under a hydrogen atmosphere (atmospheric pressure) until the reaction is complete as detected by TLC. Then filter through diatomaceous earth, concentrate the filtrate under vacuum, and purify by column chromatography to obtain compound 30B (4.67 g, yield 92.7%). MS-ESI (m / z): 306.2152 (M+H) + 。

[0492] Compound 30:

[0493]

[0494] Compound 2B (566 mg, 1.51 mmol), compound 30B (648 mg, 2.12 mmol), ethanol solution of 15% hydrogen chloride (960 mg, 3.64 mmol), and 4.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature. Then, 15 mL of saturated aqueous NaHCO3 solution was added, and a large amount of solid precipitated. After stirring for an additional 0.5 h, filtration was performed. The filter cake was washed with water and purified by slurrying with 20 mL of a mixed solvent of EA / PET = 1 / 5 to obtain compound 30 (455 mg, yield 46.9%). MS-ESI (m / z): 643.2160 (M + H) + 。 1 1H NMR (400 MHz, CDCl3) δ = 10.38 (s, 1H), 8.40 (dd, J = 8.8, 4.8 Hz, 1H), 8.22 (s, 1H), 8.08 (s, 1H), 7.48 - 7.32 (m, 2H), 7.18 (d, J = 14.0 Hz, 1H), 6.66 (s, 1H), 3.95 - 3.78 (m, 7H), 3.20 (m, 2H), 2.88 - 2.49 (m, 7H), 2.38 (s, 3H), 2.35 (s, 3H), 2.05 (d, J = 11.0 Hz, 2H), 1.90 (s, 3H), 1.86 (s, 3H), 1.88 - 1.76 (m, 2H).

[0495] Example 31

[0496] Compound 31A:

[0497]

[0498] 260 ml of DMSO was added to a 1.0 L single-necked flask. NaH (9.09 g, 226 mmol, 60%) was added portionwise under stirring, and then stirred for 10 min. Then, dimethyl malonate (26 ml, 226 mmol) was slowly added dropwise. After the addition was completed, the mixture was stirred at room temperature for 20 min, then placed at 100 °C for heat preservation reaction for 30 min - 60 min. 5-Fluoro-4-methyl-2-nitroanisole (14.0 g, 75.4 mmol) was added portionwise, and the reaction was carried out at 100 °C for heat preservation reaction for 60 min, and the reaction progress was monitored by TLC. After the reaction was completed, the reaction solution was cooled to room temperature. Then, 550 ml of water was added dropwise. After the addition was completed, the mixture was stirred at room temperature for 2 h, filtered, and the filter cake was collected and dried in a blast dryer at 50 °C to obtain compound 31A (10.5 g, yield 46.8%). MS-ESI (m / z): 298.0956 (M + H) + 。

[0499] Compound 31B:

[0500]

[0501] Dissolve Compound 31A (10.5 g, 35.32 mmol) in 300 ml of a mixed solvent (EtOH / THF = 1 / 1). After stirring for 5 min, add 150 ml of NaOH solution (2N) dropwise at room temperature. After the addition is complete, maintain the reaction at room temperature for 1 - 2 h and monitor by TLC until the reaction raw materials disappear. Then concentrate the reaction solution under reduced pressure until half of the original volume remains. Dropwise add 6 mol / L hydrochloric acid solution until the pH of the reaction solution is 2 - 3. After stirring for 10 min, add 180 ml of ethyl acetate for extraction. Extract the aqueous phase again with 100 ml of ethyl acetate. Combine the two ethyl acetate phases, wash with 100 ml of water, and then wash twice with 100 ml of saturated sodium chloride solution. Dry over anhydrous sodium sulfate, concentrate under reduced pressure, and then purify by slurry. Filter, wash the filter cake with a mixed solvent of petroleum ether∶ethyl acetate = 5∶1, and dry to obtain Compound 31B (6.4 g, yield 80.5%). MS-ESI (m / z): 226.0744 (M+H) + .

[0502] Compound 31C:

[0503]

[0504] Suspend Compound 31B (5.3 g, 23.71 mmol) in 100 ml of dichloromethane. Then cool it to 0 °C in an ice bath, add HOBT (3.53 g, 26.1 mmol), EDCI (5.0 g, 26.1 mmol), and stir at low temperature for 60 min. Then add N-methylpiperazine (2.61 g, 26.1 mmol), and then dropwise add DIPEA (3.67 g, 28.4 mmol). After the addition is complete, maintain the reaction at low temperature for 2 h, and then react at room temperature for 1 h. Monitor by TLC until the reaction raw materials disappear. After the reaction is completed, add 100 ml of water to the reaction solution, stir for 10 min, and then let it stand for phase separation. Extract the aqueous phase once again with 50 ml of dichloromethane. Combine the two dichloromethane phases, and then wash the dichloromethane phase successively with 100 ml of water, 100 ml of 3% K2CO3 aqueous solution, and 150 ml of saturated sodium chloride solution. Dry the organic phase over anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure to an oily substance, and then purify by column chromatography to obtain Compound 31C (5.9 g, yield 80.57%). MS-ESI (m / z): 308.1637 (M+H) + .

[0505] Compound 31D:

[0506]

[0507] Compound 31C (4.7 g, 15.33 mmol) was dissolved in 45 ml of anhydrous THF under nitrogen protection. After cooling to 0 °C in an ice bath, a solution of BH3·THF (45 ml, 46.02 mmol) was added dropwise. After the addition, the ice bath was removed and the mixture was stirred at room temperature overnight. The reaction was monitored by TLC until the starting materials disappeared. After the reaction was completed, 120 ml of methanol was added dropwise to quench the reaction, and then the mixture was concentrated under reduced pressure. 120 ml of methanol and 100 ml of 2N HCl solution were added to the resulting solid, and the mixture was refluxed at 70 °C for 2 h with stirring, and then concentrated under reduced pressure to about 120 ml. 2N NaOH solution was added dropwise to adjust the pH of the mixture to 8 - 9. 100 ml of ethyl acetate was added for extraction, and the aqueous phase was extracted once more with 50 ml of ethyl acetate. The two ethyl acetate phases were combined, and then the ethyl acetate phase was washed successively with 80 ml of water and 80 ml of saturated sodium chloride solution. After drying over anhydrous sodium sulfate, filtration, and concentration of the filtrate under reduced pressure to an oil, purification by column chromatography gave compound 31D (3.22 g, yield 71.50%). MS-ESI (m / z): 294.1815 (M + H) + 。

[0508] Compound 31E:

[0509]

[0510] Compound 31D (3.15 g, 10.74 mol) was dissolved in 30 ml of methanol, and 0.30 g of Pd / C (5%) was added. The mixture was purged with hydrogen 2 - 3 times and then stirred at room temperature overnight under a hydrogen atmosphere (atmospheric pressure). The reaction was monitored by TLC until the starting materials disappeared. After the reaction was completed, it was filtered through diatomaceous earth, the filtrate was concentrated, and purification by column chromatography gave compound 31E (2.55 g, yield 90.1%). MS-ESI (m / z): 264.2104 (M + H) + 。

[0511] Compound 31:

[0512]

[0513] Referring to the synthesis method of compound 2, only compound 1F therein was replaced with compound 31E to obtain compound 31. MS-ESI (m / z): 601.2084 (M + H) + 。

[0514] Example 32

[0515] Compound 32A:

[0516]

[0517] Add 1-bromo-2-fluoro-4-methoxy-5-nitrobenzene (608 mg, 2.43 mmol), 1-methyl-4-(4-piperidyl)piperazine (490 mg, 2.68 mmol), potassium carbonate (507 mg, 3.65 mmol), and 10 mL of DMF to a 100 mL round-bottom flask. Heat the mixture to 80 °C and react for 2.5 h. After detecting the completion of the reaction by TLC, cool the reaction solution to room temperature, add 30 mL of water, and a large amount of yellow solid will precipitate. Continue to stir for 1 hour and then filter by suction. Wash the filter cake with water and dry it to obtain compound 32A (743 mg, yield 74.0%).

[0518] Compound 32B:

[0519]

[0520] Add compound 32A (740 mg, 1.79 mmol), reduced iron powder (600 mg, 10.74 mmol), ammonium chloride (75 mg, 1.40 mmol), and a 67 mL ethanol / water = 3 / 1 mixed solution to a 250 mL round-bottom flask. Heat the mixture to reflux and react for about 6.5 hours until the reaction is detected to be complete by TLC. Cool the reaction solution to room temperature, adjust the pH of the reaction solution to about 8 with 2 mol / L sodium hydroxide aqueous solution, concentrate to dryness, add 110 mL of a dichloromethane / methanol = 10 / 1 mixed solution, stir at room temperature for about 0.5 hour, then filter by suction. Concentrate the filtrate and separate it by column chromatography (dichloromethane / methanol = 8 / 1) to obtain compound 32B (622 mg, yield 90.7%). MS-ESI (m / z): 383.1427 (M+H) + 。

[0521] Compound 32:

[0522]

[0523] Add compound 2B (253 mg, 0.67 mmol), compound 32B (387 mg, 1.01 mmol), a 15% hydrogen chloride ethanol solution (493 mg, 2.02 mmol), and 4 mL of ethylene glycol monomethyl ether to a reaction flask. Seal the flask and react at 100 °C for 5 - 6 h until the reaction is detected to be complete by TLC. Then cool the reaction solution to room temperature, add 4 mL of saturated NaHCO3 aqueous solution and 12 mL of H2O, and a large amount of solid will precipitate. Continue to stir for 0.5 hour and then filter by suction. Wash the filter cake with water and purify it by column chromatography (DCM / MeOH = 15 / 1 to 10 / 1), and then slurry it with 6 mL of a PE / EA = 5 / 1 mixed solvent to obtain compound 32 (173 mg, yield 35.7%). 11H NMR (400 MHz, CDCl3) δ = 10.47 (s, 1H), 8.42 - 8.26 (m, 2H), 8.21 (s, 1H), 7.53 (d, J = 8.8 Hz, 1H), 7.47 (s, 1H), 7.15 (m, 1H), 6.68 (s, 1H), 3.95 (s, 3H), 3.52 (d, J = 11.2 Hz, 2H), 2.89 - 2.51 (m, 9H), 2.41 (s, 3H), 2.39 (s, 3H), 2.21 (s, 2H), 2.05 (d, J = 11.4 Hz, 2H), 1.91 (m, 8H). MS-ESI (m / z): 722.1459 (M+H) + 。

[0524] Example 33

[0525] Compound 33A:

[0526]

[0527] Referring to the synthesis method of Reference Compound 11A, only the 2-iodo-4-fluoroaniline therein was replaced with 2-iodo-4-chloroaniline to obtain Compound 33A. MS-ESI (m / z): 204.0367 (M+H) + 。

[0528] Compound 33B:

[0529]

[0530] Referring to the synthesis method of Reference Compound 11B, only the Compound 11A therein was replaced with Compound 33A to obtain Compound 33B. MS-ESI (m / z): 393.9295 (M+H) + 。

[0531] Compound 33:

[0532]

[0533] Referring to the synthesis method of Reference Compound 11, only the Compound 11B therein was replaced with Compound 33B to obtain Compound 33. 11H NMR (400 MHz, CDCl3) δ = 10.38 (s, 1H), 8.57 - 8.32 (m, 2H), 8.05 (s, 1H), 7.47 (s, 1H), 7.32 (m, 1H), 7.18 - 6.95 (m, 1H), 6.69 (s, 1H), 3.93 (s, 3H), 3.25 (m, 2H), 2.89 - 2.53 (m, 9H), 2.42 (m, 5H), 2.21 (s, 3H), 2.05 (d, J = 11.4 Hz, 2H), 1.92 (s, 3H), 1.86 (s, 3H), 1.81 - 1.69 (m, 2H). MS-ESI (m / z): 676.1964 (M+H) + 。

[0534] Example 34

[0535] Compound 34:

[0536]

[0537] In a 25 mL round-bottom flask, weigh in compound 25C (305 mg, 0.50 mmol), K2CO3 (138 mg, 1.00 mmol), methyl iodide (142 mg, 1.00 mmol), 10 mL of DMF, stir at room temperature overnight. After the reaction is completed, purify by column chromatography to obtain compound 34 (65 mg, yield 20.5%). MS-ESI (m / z): 627.3125 (M+H) + 。

[0538] Example 35

[0539] Compound 35:

[0540]

[0541] Add compound 14B (403.27 mg, 1.19 mmol), compound 11B (500 mg, 1.32 mmol), ethanol solution of 15% hydrogen chloride (963 mg, 3.96 mmol), and 7.5 mL of ethylene glycol monomethyl ether into the reaction flask, then seal the tube and react at 100 °C for 4 h. After detecting the completion of the reaction by TLC, cool the reaction solution to room temperature, then add 15 mL of water and saturated aqueous NaHCO3 solution to adjust the pH to about 8.0, a large amount of solid precipitates, continue to stir for 0.5 h and then filter by suction. Wash the filter cake with water and dry it. The crude product is purified by slurrying with 6 mL of EA to obtain compound 35 (380 mg, yield 38%). 11H NMR (400 MHz, CDCl3) δ = 10.26 (s, 1H), 8.38 (m, 1H), 8.22 (d, J = 10.8 Hz, 2H), 7.46 - 7.32 (m, 2H), 7.03 (m, 1H), 6.62 (s, 1H), 3.88 (s, 3H), 3.41 (d, J = 11.6 Hz, 2H), 2.81 - 2.22 (m, 14H), 1.95 - 1.79 (m, 10H). MS-ESI (m / z): 680.1686 (M+H) + 。

[0542] Example 36

[0543] Compound 36A:

[0544]

[0545] In a 50 mL three-necked flask, 2-iodo-4-isopropylaniline (3.0 g, 11.5 mmol), K2CO3 (2.23 g, 16.1 mmol), Xantphos (0.174 g, 0.46 mmol), Pd(OAc)2 (0.103 g, 0.46 mmol), dimethylphosphite (1.08 g, 13.8 mmol), and 18 mL of DMF were added. The temperature was raised to 100 °C and the reaction was carried out for about 3 h until the raw materials were detected to have reacted completely by TLC. The reaction solution was cooled to room temperature, filtered by suction, 54 mL of H2O was added, a large amount of yellow solid was precipitated, filtered by suction, the filtrate was extracted with 25 mL × 3 of dichloromethane, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to obtain an oily compound 36A (2.60 g), which was directly used in the next step without purification.

[0546] Compound 36B:

[0547]

[0548] In a 50 mL single-necked flask, compound 36A (2.60 g), 5-bromo-2,4-dichloropyrimidine (2.88 g, 12.6 mmol), K2CO3 (1.91 g, 13.8 mmol), and 15 mL of DMSO were added. The temperature was raised to 60 °C and the reaction was carried out for about 3 hours until compound 36A was detected to have reacted completely by TLC. The reaction was cooled to room temperature, 45 mL of H2O was added, and it was extracted twice with 50 mL of ethyl acetate. The organic phases were combined, washed twice with 50 mL of water, washed twice with 50 mL of saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure to obtain a pink solid, and 30 mL of a mixed solvent of PE:EA = 3:1 was added for pulping. The solid obtained by filtration was dried to obtain compound 36B (2.5 g, 54%).

[0549] Compound 36:

[0550]

[0551] Compound 36B (500 mg, 1.24 mmol), compound 1F hydrochloride (484.11 mg, 1.36 mmol), and 4 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out at 100 °C for 5 - 6 h. After TLC detection showed that the reaction of compound 36B was complete, the reaction solution was cooled to room temperature. Then, 4 mL of water was added, and the pH was adjusted to about 8.0 with saturated aqueous NaHCO3. A solid was precipitated, and after stirring for an additional 0.5 h, filtration was carried out. The filter cake was washed with water and purified by column chromatography to obtain compound 36 (400 mg, yield 47.11%). 1 HNMR (400 MHz, CDCl3) δ = 10.39 (s, 1H), 8.40 (dd, J = 8.6, 4.6 Hz, 1H), 8.18 (s, 1H), 8.05 (s, 1H), 7.47 - 7.33 (m, 2H), 7.12 (dd, J = 14.4, 1.6 Hz, 1H), 6.62 (s, 1H), 3.86 (s, 3H), 3.15 (d, J = 11.6 Hz, 2H), 2.93 (m, 1H), 2.78 - 2.13 (m, 17H), 1.96 (m, 2H), 1.85 (d, J = 13.2 Hz, 6H), 1.73 (m, 2H), 1.28 (d, J = 6.8 Hz, 6H). MS-ESI (m / z): 706.2601 (M+Na) + 。

[0552] Example 37

[0553] Compound 37:

[0554]

[0555] Compound 36B (440 mg, 1.09 mmol), compound 12A (400 mg, 1.20 mmol), ethanol solution of 15% hydrogen chloride (798 mg, 3.28 mmol), and 6.0 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 100 °C for 5 - 6 h. After TLC detection showed that the reaction was complete, the reaction solution was cooled to room temperature. Then, 6 mL of saturated aqueous NaHCO3 and 12 mL of H2O were added, and a large amount of solid was precipitated. After stirring for an additional 0.5 h, filtration was carried out. The filter cake was washed with water and separated and purified to obtain compound 37 (228 mg, yield 29.8%). 11H NMR (400 MHz, CDCl3) δ = 10.34 (s, 1H), 8.35 (dd, J = 8.8, 4.8 Hz, 1H), 8.19 (s, 1H), 8.08 (s, 1H), 7.37 (m, 2H), 7.12 (d, J = 14.4 Hz, 1H), 6.67 (s, 1H), 3.86 (s, 3H), 3.08 (m, 2H), 2.92 (m, 1H), 2.80 - 2.44 (m, 11H), 2.42 - 2.26 (m, 5H), 1.95 (d, J = 11.2 Hz, 2H), 1.84 (d, J = 13.2 Hz, 6H), 1.72 (m, 2H), 1.28 (d, J = 6.8 Hz, 6H), 1.03 (t, J = 7.4 Hz, 3H). MS-ESI (m / z): 720.2757 (M+Na) + 。

[0556] Example 38

[0557] Compound 38:

[0558]

[0559] Compound 14B (309 mg, 0.91 mmol), compound 33B (300 mg, 0.76 mmol), ethanol solution of 15% hydrogen chloride (554 mg, 2.28 mmol), and 4.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out under sealed tube conditions at 100 °C for 5 - 6 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature, then 5 mL of saturated NaHCO3 aqueous solution and 10 mL of H2O were added. It was extracted with 3 × 30 mL of dichloromethane, and the organic phases were combined, washed with 30 mL of saturated NaCl aqueous solution, and the organic phase was concentrated to dryness. After separation and purification, compound 38 (154 mg, yield 29.1%) was obtained. MS-ESI (m / z): 696.1377 (M+H) + 。 1 1H NMR (400 MHz, CDCl3) δ = 10.30 (s, 1H), 8.35 (m, 1H), 8.26 (m, 2H), 7.48 - 7.30 (m, 2H), 7.05 (m, 1H), 6.68 (s, 1H), 3.89 (s, 3H), 3.38 (d, J = 11.6 Hz, 2H), 2.86 - 2.28 (m, 14H), 2.05 - 1.72 (m, 10H).

[0560] Comparative Example 1: Preparation of Control Compound 1

[0561] Control Compound 1B (intermediate):

[0562]

[0563] Referring to the synthesis method of reference compound 10B, only replace the compound 5-bromo-2,4-dichloropyrimidine therein with the compound 2,4,5-trichloropyrimidine to obtain control compound 1B.

[0564] Control compound 1:

[0565]

[0566] Referring to the synthesis method of reference compound 10, only replace the compound 10B therein with control compound 1B to obtain control compound 1. 1 H NMR (400 MHz, methanol-d4) δ = 8.02 (s, 1H), 7.93 (dd, J = 8.0, 4.0 Hz, 1H), 7.64 (s, 1H), 7.42 (t, J = 8.0 Hz, 1H), 7.15 (dd, J = 7.6, 3.6 Hz, 1H), 6.69 (s, 1H), 3.84 (s, 3H), 3.11 (d, J = 11.6 Hz, 2H), 3.02 - 2.41 (m, 13H), 2.32 (m, 4H), 2.04 (s, 3H), 2.00 (d, J = 12.6 Hz, 2H), 1.94 (s, 3H), 1.90 (s, 3H), 1.68 (m, 2H). MS-ESI (m / z): 612.2981 (M + H)+.

[0567] Comparative Example 2: Preparation of Control Compound 2

[0568] Control compound 2A (intermediate):

[0569]

[0570] Into a 100 mL three-necked flask, add (2-aminophenyl)-dimethylphosphine oxide (2.5 g, 15.00 mmol), 2,4,5-trichloropyrimidine (2.70 g, 15.00 mmol), K2CO3 (2.45 g, 67.75 mmol), nBu4NHSO4 (0.5 g, 1.50 mmol), 50 mL of DMF, heat to 65 °C and react for about 4.5 hours until the reaction is completed as detected by TLC. Subsequently, cool the reaction solution to room temperature, add 200 mL of H2O, a large amount of yellow solid precipitates, continue to stir for about 0.5 h and then filter by suction. Wash the filter cake with 100 mL of H2O and dry to obtain control compound 2A (2.84 g, yield 60.0%). MS-ESI (m / z): 316.0178 (M + H) + 。

[0571] Control compound 2:

[0572]

[0573] Compound 2A (335 mg, 1.06 mmol), compound 1F (406 mg, 1.27 mmol), ethanol solution of 15% hydrogen chloride (774 mg, 3.18 mmol), and 4.5 mL of ethylene glycol monomethyl ether were added to a reaction flask. Subsequently, the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h. After the reaction was completed as detected by TLC, the reaction solution was cooled to room temperature, then 15 mL of saturated aqueous NaHCO₃ solution was added, and a large amount of solid was precipitated. After stirring for another 0.5 h, filtration was carried out. The filter cake was washed with water, and the filter cake was purified by pulping with 9 mL of a mixed solvent of EtOH / H₂O = 1 / 2 to obtain compound 2 (320 mg, yield 50.5%). 1 ¹H NMR (400 MHz, CDCl₃) δ = 10.80 (s, 1H), 8.63 (dd, J = 8.4, 4.4 Hz, 1H), 8.10 (s, 1H), 8.04 (s, 1H), 7.51 (m, 1H), 7.38 - 7.26 (m, 2H), 7.13 (m, 1H), 6.63 (s, 1H), 3.86 (s, 3H), 3.16 (d, J = 12.0 Hz, 2H), 2.61 (m, 9H), 2.32 (s, 3H), 2.18 (s, 3H), 2.06 (m, 2H), 1.96 (m, 2H), 1.87 (s, 3H), 1.83 (s, 3H), 1.72 (m, 2H). MS-ESI (m / z): 598.2826 (M + H) + 。

[0574] Comparative Example 3: Preparation of Control Compound 3

[0575] Control Compound 3A (intermediate):

[0576]

[0577] 5-Fluoro-2-nitroanisole (2.0 g, 11.69 mmol), 1-methyl-4-(4-piperidyl)piperazine (2.57 g, 14.02 mmol), potassium carbonate (3.25 g, 23.37 mmol), and 30 mL of DMF were added to a 100 mL round-bottom flask. The temperature was raised to 120 °C and the reaction was carried out for about 3 h until the reaction was completed as detected by TLC. The reaction solution was cooled to room temperature, 30 mL of water was added, and extraction was carried out with 30 mL × 3 of ethyl acetate. The organic phases were combined, the organic phase was washed with 30 mL of saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain control compound 3A (3.57 g, yield 91.3%).

[0578] Control Compound 3B (intermediate):

[0579]

[0580] Compound 3A (3.57 g, 10.68 mmol), 0.36 g of 5% Pd / C, 72 mL of methanol were purged with hydrogen 2 - 3 times, and stirred overnight at room temperature under a hydrogen atmosphere (atmospheric pressure) until the reaction was completed as detected by TLC. Subsequently, after filtration through diatomaceous earth, the filtrate was concentrated in vacuo, and purified by column chromatography to obtain Compound 3B (2.17 g, yield 66.8%). MS-ESI (m / z): 305.2315 (M + H) + 。

[0581] Compound 3:

[0582]

[0583] Compound 1B (400 mg, 1.21 mmol), Compound 3B (443 mg, 1.45 mmol), ethanol solution of 15% hydrogen chloride (884 mg, 3.63 mmol), and 6 mL of ethylene glycol monomethyl ether were added to a reaction flask, and the reaction was carried out in a sealed tube at 120 °C for 5 - 6 h until the reaction was completed as detected by TLC. Subsequently, the reaction solution was cooled to room temperature, 6 mL of saturated aqueous NaHCO3 solution and 6 mL of H2O were added, extracted with 30 mL × 3 of dichloromethane, the combined organic phases were washed with 20 mL of saturated aqueous sodium chloride solution, dried over anhydrous sodium sulfate, filtered, concentrated, and purified by column chromatography (DCM / MeOH = 8 / 1). After slurrying with 5 mL of a mixed solvent of PE / EA = 4 / 1, Compound 3 (257 mg, yield 35.4%) was obtained. 1 1H NMR (400 MHz, CDCl3) δ = 10.63 (s, 1H), 8.49 (dd, J = 8.8, 4.8 Hz, 1H), 8.19 - 7.99 (m, 2H), 7.31 (d, J = 8.8 Hz, 1H), 7.25 (s, 1H), 7.07 (d, J = 14.0 Hz, 1H), 6.61 - 6.44 (m, 2H), 3.87 (s, 3H), 3.66 (d, J = 12.0 Hz, 2H), 2.66 (m, 9H), 2.38 (s, 3H), 2.32 (s, 3H), 2.21 (s, 2H), 1.97 (m, 2H), 1.85 (s, 3H), 1.81 (s, 3H), 1.73 (m, 2H). MS-ESI (m / z): 598.2804 (M + H) + 。

[0584] Comparative Example 4: Preparation of Compound 4

[0585] Compound 4A (intermediate):

[0586]

[0587] Into a 1 L three-necked flask, add 2-iodo-4-bromoaniline (30 g, 100.7 mmol), K2CO3 (22.27 g, 161.1 mmol), Xantphos (5.83 g, 10.1 mmol), Pd(OAc)2 (1.12 g, 4.9 mmol), dimethylphosphite (11.78 g, 150.9 mmol), 450 mL of DMF. Under N2 protection, heat to 100 °C and react for about 3 h until the reaction is completed as detected by TLC. Subsequently, cool the reaction to room temperature, add 1350 mL of H2O dropwise, stir and then filter by suction. Wash the filter cake with water. The filtrate is extracted with 500 mL of dichloromethane three times. Combine the organic phases. Wash the combined organic phases with 500 mL of saturated sodium chloride aqueous solution twice. Dry over anhydrous sodium sulfate, filter and concentrate to obtain reference compound 4A (14 g, yield 56%).

[0588] Reference compound 4B (intermediate):

[0589]

[0590] Into a 500 mL three-necked flask, add compound 4A (12 g, 48.4 mmol), cyclopropylboronic acid (8.31 g, 96.7 mmol), K2CO3 (13.36 g, 96.7 mmol), Pd(dppf)Cl2 (1.77 g, 2.42 mmol), 1,4-dioxane (120 mL), H2O (12 mL). Under N2 protection, heat to 100 °C and react for about 8 h until the reaction is completed as detected by TLC. Subsequently, cool the reaction to room temperature, add 150 mL of H2O, stir and then filter by suction. Wash the filter cake with dichloromethane. Let it stand for layering. Extract the aqueous phase with 150 mL of dichloromethane three times. Combine the organic phases. Dry over anhydrous sodium sulfate, filter and concentrate, and then purify by column chromatography to obtain reference compound 4B (7.5 g, yield 74%).

[0591] Reference compound 4C (intermediate):

[0592]

[0593] Into a 100 mL reaction flask, add compound 4B (7.5 g, 35.8 mmol), K2CO3 (5.95 g, 43.1 mmol), 5-bromo-2,4-dichloropyrimidine (8.99 g, 39.5 mmol), DMSO (45 mL). Heat the mixture to 60 °C and react for about 2.5 h until the reaction is completed as detected by TLC. Subsequently, cool the reaction to room temperature, add dropwise 135 mL of H2O, a large amount of brownish-yellow solid precipitates. Continue stirring for 1 h and then filter. Wash the filter cake with 100 mL of H2O, and dry the filter cake to obtain the control compound 4C (7.4 g, yield 51.6%).

[0594] Control compound 4:

[0595]

[0596] Into a 100 mL reaction flask, add compound 4C (1.7 g, 4.24 mmol), compound 12A (1.55 g, 4.66 mmol), trifluoroacetic acid (1.21 g, 10.6 mmol), ethylene glycol monomethyl ether (27 mL). Heat the mixture to 100 °C and react for about 7 h until the reaction is completed as detected by TLC. Subsequently, cool the reaction to room temperature, add dropwise 54 mL of H2O, and then add dropwise 27 mL of saturated NaHCO3 aqueous solution. A large amount of solid precipitates. Continue stirring for 1 h and then perform suction filtration. Wash the filter cake with water. Pulverize and purify the filter cake with 15 mL of ethyl acetate, and then filter and dry to obtain the control compound 4 (1.68 g, yield 56.8%). 1 1H NMR (400 MHz, CD3OD) δ = 8.13 (s, 1H), 7.88 (dd, J = 8.8, 4.8 Hz, 1H), 7.66 (s, 1H), 7.43 (dd, J = 14.0, 2.2 Hz, 1H), 7.19 (m, 1H), 6.76 (s, 1H), 3.85 (s, 3H), 3.04 (m, 2H), 2.32 - 2.75 (m, 16H), 2.01 (m, 3H), 1.83 (m, 6H), 1.74 - 1.65 (m, 2H), 1.06 (m, 2H), 0.96 (t, J = 7.6 Hz, 3H), 0.76 (m, 2H). MS-ESI (m / z): 696.2711 (M + H) + 。

[0597] Experimental Example 1 Enzyme Activity Experiment 1

[0598] 1. Experimental Purpose: To detect the effect of the test compound on EGFR kinase activity

[0599] 2. Experimental Information:

[0600] To evaluate the effect of the compound on the activities of kinase EGFR wild-type and mutants, the ADP-Glo (Promega) kit was used to measure the ATP consumption in the enzymatic reaction to detect the kinase activity. The test sample was dissolved in DMSO and serially diluted. In a microplate, EGFR kinase, reaction buffer (containing Tris-HCl pH 7.5, MgCl2, DTT, and BSA), kinase substrate Poly(Glu4,Tyr1), and the sample (total volume per well 20 μL) were added to each well, and a blank control (without enzyme and sample) and a negative control (without sample) were set up simultaneously; incubated at 23 °C for 15 min; 5 μL of ATP was added, and the reaction was carried out at 23 °C for 60 min; ADP-Glo Reagent was added, and the reaction was continued at room temperature for 40 min to inactivate the excess ATP; then kinase detection reagent was added, and after reacting at room temperature for 30 min, the chemiluminescence intensity L of each well was measured. The inhibition rate of the compound was calculated according to the chemiluminescence intensity L value, and the inhibition rate = [1 - (L 样品 - L 空白 ) / (L 阴性 - L 空白 )] × 100%. According to the above calculation, the 4Parameter Logistic Model in XLfit software was used to calculate the IC 50 value.

[0601] 3. Test results: The IC 50 values of the compounds of the examples of the present invention and the compounds of the comparative examples for the enzymatic activities of EGFR-Del19, EGFR-C797S / Del19, EGFR-T790M / Del19, EGFR-T790M / L858R, EGFR-C797S / T790M / L858R, EGFR-C797S / T790M / Del19, and EGFR-WT are shown in Table 1 and Table 2.

[0602] 4. Conclusion: As can be seen from Table 1 and Table 2, the compounds of the examples of the present invention have good selectivity for the enzymatic activity of EGFR wild-type (EGFR-WT), and have good inhibitory effects on the enzymatic activities of EGFR single mutation, double mutation, and triple mutation (such as EGFR-Del19, EGFR-C797S / Del19, EGFR-T790M / Del19, EGFR-T790M / L858R, EGFR-C797S / T790M / L858R, EGFR-C797S / T790M / Del19), all of which are much better than the compounds of the comparative examples.

[0603] Table 1

[0604]

[0605] Table 2

[0606]

[0607] Experimental Example 2 Enzyme Activity Experiment 2

[0608] 1. Experimental Purpose: To detect the effect of the test compound on ALK kinase activity

[0609] 2. Experimental Information:

[0610] To evaluate the effect of the compound on the kinase EML4-ALK activity, the ADP-Glo (Promega) kit was used to measure the ATP consumption in the enzymatic reaction to detect the kinase activity. The test sample was dissolved in DMSO and serially diluted. In the microplate, each well was added with EML4-ALK kinase, reaction buffer (containing Tris-HCl pH 7.5, MgCl2, DTT and BSA), kinase substrate IGF1 and sample (total volume per well 20 μL), and at the same time, a blank control (without enzyme and sample) and a negative control (without sample) were set up; 5 μL of ATP was added and reacted at 23 °C for 60 min; ADP-Glo Reagent was added and the reaction was continued at room temperature for 40 min to inactivate the excess ATP; then the kinase detection reagent was added, and after reacting at room temperature for 30 min, the chemiluminescence intensity L of each well was measured. According to the chemiluminescence intensity L value, the inhibition rate of the compound was calculated, and the inhibition rate = [1 - (L 样品 - L 空白 ) / (L 阴性 - L 空白 )] × 100%. According to the above calculation, the 4Parameter Logistic Model in XLfit software was used to calculate the IC 50 value.

[0611] 3. Test Results: The IC 50 values of the compounds of the examples of the present invention and the compounds of the comparative examples for ALK kinase activity are shown in Table 3.

[0612] 4. Conclusion: As can be seen from Table 3, the compounds of the examples of the present invention have a good inhibitory effect on the enzyme activity of ALK kinase, and are all superior to the compounds of the comparative examples.

[0613] Table 3

[0614]

[0615]

[0616] Experimental Example 3 Cell Anti-Proliferation Experiment 1

[0617] 1. Experimental purpose: To detect the effect of the test compound on the proliferation of BaF3 cell lines with EGFR mutations

[0618] 2. Experimental information:

[0619] 2.1. Cell information: The BaF3 cell line with EGFR mutations is from WuXi AppTec:

[0620] BaF3 cell line expressing EGFR mutations

[0621] EGFR-T790M / Del19

[0622] EGFR-C797S / Del19

[0623] EGFR-T790M / L858R

[0624] EGFR-C797S / L858R

[0625] EGFR-C797S / T790M / L858R

[0626] EGFR-C797S / T790M / Del19

[0627] EGFR-WT wild-type cell line

[0628] 2.2. Experimental method

[0629] Day 0: Seeding plates

[0630] a. Turn on the ultraviolet lamp of the biosafety cabinet and set a 30-minute countdown.

[0631] b. Preheat the culture medium in a 37°C water bath.

[0632] c. After the ultraviolet irradiation is completed, turn on the biosafety cabinet. Wipe the preheated culture medium, PBs, etc. with alcohol and place them in the biosafety cabinet.

[0633] d. Take out the cells from the incubator, pipette them into a uniform cell suspension in the biosafety cabinet, and then count.

[0634] e. According to the cell counting results, adjust the cell suspension density to 3000 cells per well, and seed 50 μL per well into a 384-well plate.

[0635] f. Incubate the seeded cells in an incubator at 37°C and 5% CO2 for 3 hours.

[0636] g. Use Tecan (liquid workstation) to add the compound to the cell plate.

[0637] Day 3: Equilibrate the cell plates with added compounds and CTG (CellTiter-Glo luminescent cell viability assay reagent) at room temperature. Then add 25 μl of CTG to each well, centrifuge at 1000 rpm for 1 min, and shake for 1 - 2 min. Centrifuge again at 1000 rpm for 1 min, let stand for 10 min, and then detect the signal value in Envision. Use XL-Fit analysis software to calculate the IC 50 (The concentration of the compound corresponding to the 50% inhibition rate is the IC of the compound's inhibition of cell viability 50 ). Inhibition rate % = (reading of the control group without drug - reading of the sample) / reading of the control group without drug × 100

[0638] 3. Test results: The IC of the compounds in the examples of the present invention, the compounds in the comparative examples, and the main drugs brigatinib and osimertinib for treating non-small cell lung cancer on the market for inhibiting the activity of Ba / F3 cells expressing EGFR-T790M / Del19, EGFR-C797S / Del19, EGFR-T790M / L858R, EGFR-C797S / L858R, EGFR-C797S / T790M / L858R, EGFR-C797S / T790M / Del19, and EGFR-WT 50 values are shown in Tables 4 and 5.

[0639] 4. Conclusion: From Tables 4 and 5, we can see that the compounds in the examples of the present invention have good selectivity for the activity of Ba / F3 cells expressing EGFR wild type (EGFR-WT), and have good inhibitory effects on the proliferation of Ba / F3 cells expressing EGFR double mutations and triple mutations (such as EGFR-T790M / Del19, EGFR-C797S / Del19, EGFR-T790M / L858R, EGFR-C797S / L858R, EGFR-C797S / T790M / L858R, EGFR-C797S / T790M / Del19). In particular, the inhibitory activities of compounds 1, 2, 4, 5, 6, 9, 11, 12, 14, 23, 24, 25, 28, 29, 30, 32, 33, 35, 36, 37, 38 in the examples on Ba / F3 cell lines expressing EGFR double mutations and triple mutations are much better than those of control compounds 1, 2, 3 and the marketed drugs brigatinib and osimertinib.

[0640] Table 4

[0641]

[0642] Table 5

[0643]

[0644] Experimental Example 4 Cell Anti - proliferation Experiment 2

[0645] 1. Experimental Purpose: To detect the effect of the test compound on the proliferation of PC9 cell line with EGFR mutation

[0646] 2. Experimental Information:

[0647] 2.1. Cell Information: PC9 cells carry EGFR Del19 mutation, and the remaining mutant cells are constructed from PC9 cells according to the common stable cell line construction method.

[0648] PC9 - cell line expressing EGFR mutation

[0649] EGFR - Del19

[0650] EGFR - T790M / Del19

[0651] EGFR - C797S / T790M / Del19

[0652] 2.2. Experimental Method:

[0653] The test compound was used as the starting concentration for the experiment at an appropriate concentration, diluted 5 - fold each time, and a total of 6 concentration gradients were diluted. Brigatinib (purchased from Selleck) was used as the starting concentration for the experiment at 5 μM, diluted 3 - fold each time, and a total of 6 concentration gradients were diluted. The test compound and brigatinib were respectively added to the above - mentioned cells, incubated at 37 °C and 5% CO2 for 72 hours. The SRB (Sulforhodamine B) detection method was used, and the optical density value of each well was read at a wavelength of 490 nm with an enzyme - linked immunosorbent assay reader.

[0654] The optical density value of the cells at the time of 0 drug action was set as the Tz value, representing the value of the cells when the drug was added. The optical density value of the cells treated with the solvent control DMSO for 72 hours was set as the C value; the optical density value of the cells treated with the test compound for 72 hours was set as the Ti value. According to the method proposed by the US NIH - NCI (National Institutes of Health - National Cancer Institute), the response of the cells to the drug was calculated as follows: when Ti ≥ Tz, it was [(Ti - Tz) / (C - Tz)]×100; when Ti < Tz, it was [(Ti - Tz) / Tz]×100. According to the above calculations, the GI50 value (the concentration of the test compound required when the cell growth inhibition rate is 50%) was calculated using the 4 - Parameter Logistic Model in XLfit software.

[0655] 3. Test Results: The GI values of the compounds of the embodiments of the present invention, the compounds of the comparative embodiments, and the main drug brigatinib for treating non-small cell lung cancer on the market against PC9 cells expressing EGFR-Del19, EGFR-T790M / Del19, and EGFR-C797S / T790M / Del19 are shown in Table 6. 50 The values are shown in Table 6.

[0656] 4. Conclusion: As can be seen from Table 6, the compounds of the embodiments of the present invention have a good inhibitory effect on the proliferation activity of PC9 cells expressing EGFR single mutation, double mutation, and triple mutation (such as EGFR-Del19, EGFR-T790M / Del19, and EGFR-C797S / T790M / Del19), and the inhibitory activity is much better than that of control compounds 1, 2, 3, and the marketed drug brigatinib.

[0657] Table 6

[0658]

[0659] Experimental Example 5 Cell Anti-Proliferation Experiment 3

[0660] 1. Experimental Purpose: To detect the effect of the test compound on the proliferation of BaF3 cell lines containing ALK mutations

[0661] 2. Experimental Information:

[0662] 2.1. Cell Information: The BaF3 cell lines containing ALK mutations are from WuXi AppTec.

[0663] Ba / F3 - cell lines expressing ALK gene fusion and mutations

[0664] Ba / F3-EML-4-ALK-WT

[0665] Ba / F3-EML-4-ALK-L1196M

[0666] 2.2. Experimental Method

[0667] Day 0: Seeding plates

[0668] a. Turn on the ultraviolet lamp of the biosafety cabinet and count down for 30 min.

[0669] b. Preheat the culture medium in a 37-degree water bath.

[0670] c. After the ultraviolet irradiation is completed, turn on the biosafety cabinet. Wipe the preheated culture medium, PBS, etc. with alcohol and put them into the biosafety cabinet.

[0671] d. Take out the cells from the incubator, pipette them into a uniform cell suspension in the biosafety cabinet and then count.

[0672] e. According to the cell counting results, adjust the cell suspension density to 3000 cells per well, and seed 50 μL per well into a 384-well plate.

[0673] f. Incubate the seeded cells in an incubator at 37 °C and 5% CO2 for 3 hours.

[0674] g. Use Tecan (liquid workstation) to add the compound to the cell plate.

[0675] Day 3: Equilibrate the cell plate with the added compound and CTG at room temperature. Then add 25 μL of CTG to each well, centrifuge at 1000 rpm for 1 min, and shake for 1 - 2 min. Centrifuge again at 1000 rpm for 1 min, let stand for 10 min, and then detect the signal value in Envision. Use XL-Fit analysis software to calculate the IC 50 (The concentration of the compound corresponding to the 50% inhibition rate is the IC of the compound's inhibition of cell activity 50 ). Inhibition rate % = (reading of the control group without drug - reading of the sample) / reading of the control group without drug × 100

[0676] 3. Test results: The IC 50 values of the compounds in the examples of the present invention and the compounds in the comparative examples for BaF3 cells of EML-4-ALK-WT and EML-4-ALK-L1196M are shown in Table 7.

[0677] 4. Conclusion: As can be seen from Table 7, the compounds in the examples of the present invention have very good inhibitory effects on the proliferation of BaF3 cells expressing EML-4-ALK-WT and EML-4-ALK-L1196M, and the inhibitory activity is superior to that of control compounds 1, 2, and 3.

[0678] Table 7

[0679]

[0680] Experimental Example 6: In vivo pharmacodynamic study on an engineered BaF3 EGFR-DTC (C797S / T790M / Del19) cell nude mouse xenograft model

[0681] The model used in this experiment was an engineered subcutaneous xenograft tumor model of BaF3 EGFR-DTC (C797S / T790M / Del19) cells in BALB / c nude mice. The engineered BaF3 cells were cultured in suspension in vitro. The culture conditions for BaF3 EGFR-DTC (C797S / T790M / Del19) were RPMi-Compound 140 Medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomycin, 10 μg / ml Blastcidin, and cultured in an incubator at 37 °C with 5% CO2. Passage was performed twice a week. When the cell number reached the requirement, the cells were harvested, counted, and inoculated. 0.2 ml (10 6 cells) (added with Matrigel, volume ratio 1:1) were subcutaneously inoculated into the right back of each mouse. When the average tumor volume reached 109 mm 3 , grouping and drug administration were started, with 6 mice in each group.

[0682] Route and frequency of drug administration: Oral gavage, drug administration volume 10 ml / kg, and the model group was given the same volume of solvent. Administer the drug once a day for 14 consecutive days. The doses of Compound 2 and Brigatinib were 25 mg / kg and 50 mg / kg, and the dose of Compound 33 was 25 mg / kg.

[0683] The general conditions of the mice such as spirit, activity, and food intake were observed daily, and the body weight was measured 3 times a week; the short diameter (a) and long diameter (b) of the tumors of each mouse were measured 3 times a week with a vernier caliper, and the tumor volume was calculated according to the formula (a 2 ×b) / 2. The relative tumor volume (RTV) was calculated based on the measured tumor volume, RTV = V t / V0. Where V0 is the tumor volume at random grouping (i.e., d0), and V t is the tumor volume at each measurement (i.e., d n ). The relative tumor proliferation rate, an evaluation index of antitumor activity, was calculated according to the following formula: Relative tumor proliferation rate T / C (%)

[0684]

[0685] (Note: T RTV : treatment group RTV ; C RTV : RTV of the model control group. According to the efficacy evaluation criteria in the "Technical Guidelines for Non-Clinical Research of Cytotoxic Antitumor Drugs" issued by the China National Medical Products Administration, T / C% ≤ 40% is considered effective).

[0686] The results of the effects of each test sample on the tumor situation of the BaF3 EGFR-DTC (C797S / T790M / Del19) cell nude mouse xenograft tumor model are shown in Table 8-9;

[0687] Table 8: Average tumor volume (mm 3 )

[0688]

[0689] Note: a. Days after administration;

[0690] ▲ : In the model control group, 3 mice died on the 10th day, 2 mice died on the 12th day, and 1 mouse died on the 13th day after administration. Autopsy showed that the death was due to tumor metastasis;

[0691] Table 9: Relative tumor proliferation rate T / C (%) of BaF3 EGFR-DTC (C797S / T790M / Del19) nude mouse xenograft tumor model

[0692]

[0693] ★★ : Compared with the original RTV data of the same-dose group of Compound 2 at the same time point, p < 0.01;

[0694] ☆☆ : Compared with the original RTV data of the 33 (25 mg / kg) dose group of Compound 3 at the same time point, p < 0.01;

[0695] ▲ : Since all the animals in the model control group died on the 13th day after administration, the calculation of T / C (%) was up to the 12th day after administration.

[0696] The above test results show that: in this experiment, Compound 2 at doses of 25 mg / kg and 50 mg / kg, and Compound 33 at a dose of 25 mg / kg could significantly inhibit the tumor growth of BaF3 EGFR-DTC (C797S / T790M / Del19) nude mouse xenograft tumors; after 12 days of administration, the T / C% of Compound 2 (25 mg / kg) and Compound 2 (50 mg / kg) were 3.12% and 1.91% respectively, and the T / C% of Compound 33 (25 mg / kg) was 2.56%; while Brigatinib showed no obvious inhibitory effect on the tumor growth of BaF3 EGFR-DTC (C797S / T790M / Del19) nude mouse xenograft tumors. After 12 days of administration, the T / C% of Brigatinib (25 mg / kg) and Brigatinib (50 mg / kg) were 84.64% and 41.67% respectively.

[0697] It can be seen that the inhibitory effect of Compound 2 on tumor growth is significantly stronger than that of Brigatinib. On the 12th day of administration, the T / C% of Compound 2 and Brigatinib (25 mg / kg: 3.12% vs. 84.64%; 50 mg / kg: 1.91% vs. 41.67%) showed significant differences (p < 0.01). In addition, the inhibitory effect of Compound 33 (25 mg / kg) on tumor growth was also significantly stronger than that of Brigatinib, and on the 12th day of administration, the difference in T / C% between Compound 33 and Brigatinib was significant (p < 0.01).

[0698] The above results suggest that the inhibitory effects of Compound 2 and Compound 33 on the growth of xenograft tumors in BaF3 EGFR-DTC (C797S / T790M / Del19) nude mice are significantly stronger than that of Brigatinib.

[0699] Experimental Example 7: In Vivo Pharmacodynamic Study on the Xenograft Tumor Model of Engineered BaF3 EML-4-ALK-L1196M Cells in Nude Mice

[0700] The model used in this experiment was the subcutaneous xenograft tumor model of engineered BaF3 EML-4-ALK-L1196M cells in BALB / c nude mice. The engineered BaF3 cells were cultured in suspension in vitro. The culture conditions for BaF3 EML-4-ALK-L1196M were RPMI-Compound 140 Medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, 100 μg / ml streptomycin, 10 μg / ml Blastcidin, and cultured in an incubator at 37°C with 5% CO2. Passage was performed twice a week. When the cell number reached the required amount, the cells were harvested, counted, and inoculated. 0.2 ml (10 6 cells) (added with Matrigel, volume ratio 1:1) were subcutaneously inoculated into the right back of each mouse. When the average tumor volume reached 128 mm 3 , grouping and administration began, with 6 mice in each group.

[0701] Administration method and frequency: Oral gavage, administration volume 10 ml / kg, and the model group was given the same volume of solvent. Administered once a day for 14 consecutive days. The dose of Compound 2 was 50 mg / kg, the dose of Compound 6 was 60 mg / kg, and the dose of Compound 14 was 80 mg / kg.

[0702] The general conditions of the mice such as spirit, activity, and food intake were observed every day, and the body weight was measured 3 times a week; the short diameter (a) and long diameter (b) of the tumors of each mouse were measured 3 times a week with a vernier caliper, and the tumor volume was calculated according to the formula (a 2 ×b) / 2. The relative tumor volume (RTV) was calculated based on the measured and calculated tumor volume, RTV = V t / V0. Where V0 is the tumor volume at random grouping (i.e., d0), and V t is the tumor volume at each measurement (i.e., d n ). Calculate the relative tumor proliferation rate, an evaluation index of anti-tumor activity, according to the following formula: Relative Tumor Proliferation Rate T / C (%)

[0703]

[0704] (Note: T RTV : Treatment group RTV ; C RTV : Model control group RTV. According to the evaluation criteria for efficacy in the "Technical Guidelines for Non-Clinical Research of Cytotoxic Anti-Tumor Drugs" issued by the China National Medical Products Administration, T / C% ≤ 40% is considered effective).

[0705] The results of the effects of each test sample on the tumor situation of the BaF3 EML-4-ALK-L1196M cell nude mouse xenograft model are shown in Table 10-11;

[0706] Table 10: Average tumor volume (mm 3 ) of each group at each time point in the BaF3 EML-4-ALK-L1196M nude mouse xenograft model

[0707]

[0708] Note: a Days after administration.

[0709] Table 11: Relative Tumor Proliferation Rate T / C (%) in the BaF3 EML-4-ALK-L1196M Nude Mouse Xenograft Model

[0710]

[0711] The above test results show that: In this experiment, Compound 2 (50 mg / kg), Compound 6 (60 mg / kg), and Compound 14 (80 mg / kg) can all significantly inhibit the tumor growth of the BaF3 EML-4-ALK-L1196M nude mouse xenograft.

[0712] Experimental Example 8: Single-dose Toxicity Test Study of Compound 2 and Brigatinib Administered Orally by Gavage to SD Rats

[0713] SD rats at 8 weeks old, 10 rats in each group, with 5 males and 5 females. The animals were purchased from Beijing Spevogen. The animals were acclimated in this laboratory for 3 days before the experiment. Oral gavage was performed, and the administration volume was 10 ml / kg. The rats were fasted for 17 h before administration without water restriction. The control group rats were orally gavaged with an equal volume of vehicle. Food was restored about 2 hours after the administration ended.

[0714] Observe twice a day after administration. The day of the first administration is defined as day 1 of the trial, observe continuously for 14 days, and the experiment ends on day 15.

[0715] The experimental doses, groups, and the death situation of animals on the 15th day after administration are shown in Table 12:

[0716] Table 12: Death situation of animals in the single-dose toxicity test of compound 2 and Brigatinib administered orally to SD rats

[0717]

[0718] The above test results show that: in the single-dose toxicity test of compound 2 and Brigatinib administered orally to SD rats, the maximum tolerated dose (MTD) of compound 2 is 125 mg / kg, while the MTD of Brigatinib < 75 mg / kg; it is suggested that under the conditions of this test, the single-dose toxicity of compound 2 in SD rats is much lower than that of Brigatinib, and it has better safety.

[0719] Experimental Example 9: In vivo pharmacodynamic study on a BALB / c nude mouse model with subcutaneous xenograft tumors of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cells subcutaneously inoculated on both sides

[0720] The model used in this test is a BALB / c nude mouse subcutaneous xenograft tumor model with NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cells subcutaneously inoculated on both sides of the same BALB / c nude mouse (note: NCI-H3122 is an EML4-ALK fusion gene positive cell line).

[0721] The culture conditions of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) are both in RPMI-1640 medium supplemented with 10% fetal bovine serum, 100 U / ml penicillin, and 100 μg / ml streptomycin, and cultured in a 5% CO2 incubator at 37 °C. Passage is carried out twice a week. When the cell number reaches the requirement, the cells are harvested, counted, and inoculated.

[0722] When inoculating, 0.2 ml (5x10 6 cells) of NCI-H3122 cells (added with Matrigel, volume ratio 1:1) are subcutaneously inoculated on the left lower abdomen of each mouse, and 0.2 ml (5 × 10 6One NCI-H1975 EGFR DTC (C797S / T790M / Del19) cell (mixed with Matrigel at a volume ratio of 1:1) was subcutaneously inoculated into the right lower abdomen of each mouse. When the average volume of the NCI-H3122 subcutaneous xenograft tumor reached approximately 116 mm 3 , the average volume of the NCI-H1975 EGFR DTC (C797S / T790M / Del19) subcutaneous xenograft tumor reached approximately 110 mm 3 , grouping and drug administration were started, with 8 mice in each group.

[0723] Route and frequency of drug administration: Oral gavage, drug administration volume 10 mL / kg, and the model group was given the same volume of solvent. Administer the drug once a day for 16 consecutive days. The doses of Compound 2 were 20 mg / kg and 40 mg / kg, and the dose of Crizotinib was 50 mg / kg.

[0724] The general conditions of the mice such as spirit, activity, and food intake were observed daily, and the body weight was measured 3 times a week; the short diameter (a) and long diameter (b) of the tumors of each mouse were measured with a vernier caliper 3 times a week, and the tumor volume was calculated according to the formula (a 2 ×b) / 2. The relative tumor volume (RTV) was calculated based on the measured tumor volume, RTV = V t / V0. Where V0 is the tumor volume at random grouping (i.e., d0), and Vt is the tumor volume at each measurement (i.e., d n ). Calculate the relative tumor proliferation rate, an evaluation index of anti-tumor activity, according to the following formula: Relative tumor proliferation rate T / C (%)

[0725]

[0726] Note: T RTV : Treatment group RTV ; C RTV : RTV of the model control group. According to the evaluation criteria for efficacy in the "Technical Guidelines for Non-Clinical Research of Cytotoxic Antitumor Drugs" issued by the China National Medical Products Administration, T / C% ≤ 40% is considered effective.

[0727] The results of the effects of each test sample on the tumor conditions of the BALB / c nude mouse model with subcutaneous xenograft tumors of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cells subcutaneously inoculated on both sides of the same BALB / c nude mouse are shown in Tables 13 and 14.

[0728] Table 13: Average tumor volume of each group of BALB / c nude mice models with subcutaneous transplanted tumors inoculated subcutaneously with NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cells on both sides at different time points

[0729]

[0730] Note: a Days after administration

[0731] Table 14: Relative tumor proliferation rate T / C (%) of BALB / c nude mice models with subcutaneous transplanted tumors inoculated subcutaneously with NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cells on both sides

[0732]

[0733] The above test results show that: in this test, at doses of 20 mg / kg and 40 mg / kg, compound 2 can significantly inhibit the tumor growth of subcutaneous transplanted tumors of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cells in nude mice, and there is a dose-effect relationship; 16 days after administration, the T / C% of compound 2 (20 mg / kg) for subcutaneous transplanted tumors of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) are 8.16% and 37.95% respectively, and the T / C% are both < 40%, reaching the evaluation criteria for efficacy in the "Technical Guidelines for Non-Clinical Research of Cytotoxic Antitumor Drugs" issued by the China National Medical Products Administration; while the T / C% of compound 2 (40 mg / kg) for subcutaneous transplanted tumors of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) are 3.6% and 14.36% respectively, with a stronger effect on inhibiting tumor growth and a clear dose-effect relationship;

[0734] However, Crizotinib (50 mg / kg) did not show an obvious effect on inhibiting the tumor growth of subcutaneous transplanted tumors of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cells in nude mice in this test. 16 days after administration, the T / C% of subcutaneous transplanted tumors of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) are 49.35% and 95.67% respectively, and neither reached the effective standard.

[0735] The above results suggest that compound 2 can significantly inhibit the tumor growth of NCI-H3122 and NCI-H1975 EGFR DTC (C797S / T790M / Del19) cell xenografts in nude mice, and there is a clear dose-effect relationship.

[0736] Experimental Example 10 Cell Anti-Proliferation Experiment 4

[0737] 1. Experimental Purpose: To detect the effect of the test compound on the proliferation of BaF3 cells with ALK mutations

[0738] 2. Experimental Materials

[0739] BaF3 EML-4-ALK-L1196M cell line, brand: Kangyuan Bochuang, catalog number: KC-0102

[0740] Cell Counting Kit-8 (CCK-8), brand: Targetmol, catalog number: C0005

[0741] Multifunctional microplate reader POLARstar Omega, brand: BMG LABTECH

[0742] 3. Experimental Method

[0743] Day 0: Seeding

[0744] a. Take out the cells from the incubator, pipette them into a uniform cell suspension in the laminar flow hood and count.

[0745] b. According to the cell counting results, adjust the cell suspension density and inoculate 110 μL per well (containing 8000 cells) into a 96-well plate.

[0746] c. Add 10 μL of the gradient-diluted compound to the cell plate to make the final concentration of the compound 500, 166.67, 55.56, 18.52, 6.17, 2.06, 0.69, 0.23, 0.08 nM.

[0747] d. Incubate the cell plate in an incubator at 37 °C and 5% CO2 for 72 h.

[0748] Day 3: After equilibrating the CCK8 to room temperature, add 10 μL of CCK8 to each well. Incubate the cell plate in an incubator at 37 °C and 5% CO2 for 2 h, and then detect the signal value OD in the multifunctional microplate reader POLARstar Omega. 450 . Use GraphPadprism analysis software to calculate the IC of each compound by computer fitting 50 (The concentration of the compound corresponding to 50% inhibition rate is the IC of the compound's inhibition of cell activity50 )。Inhibition rate % = (Reading of the control group without drug - Reading of the sample) / (Reading of the control group without drug - Reading of the control group with only culture medium) × 100

[0749] 4. Test results: The IC 50 values of the compounds in the examples of the present invention and the compounds in the comparative examples for BaF3 cells expressing EML-4-ALK-L1196M are shown in Table 15.

[0750] Conclusion: As can be seen from Table 15, the compounds in the examples of the present invention have a very good inhibitory effect on the proliferation of BaF3 cells expressing EML-4-ALK-L1196M, and the inhibitory activity is superior to that of control compound 4.

[0751] Table 15: Inhibitory effects of compound 2 and control example 4 on BaF3 EML-4-ALK-L1196M cells

[0752] Compound <![CDATA[First test IC 50 (nM)]]> <![CDATA[Second test IC 50 (nM)]]> Compound 2 15.14 10.38 Control Compound 4 81.45 74.61

[0753] Experimental example 11 Pharmacokinetic experiment

[0754] Six SD rats, all male, about 8 - 9 weeks old, with a body weight of about 430 g / rat, were fasted for 18 hours without water deprivation.

[0755] They were divided into two groups, with 3 rats in each group. The rats in group 1 were administered with compound 2, and the rats in group 2 were administered with control compound 4, both by oral gavage, with a dosing dose of 20 mg / kg. The gastric lavage solution was prepared using 70 mM citric acid buffer (pH 3.0), and the dosing volume was 5 ml / kg.

[0756] Blood samples of 0.6 ml (anticoagulated with EDTA-2K) were taken from the rat orbital cavities at 0.5, 1.5, 3, 4.5, 8, 12, 24, 36, 48, and 72 h after dosing respectively. The plasma was separated and stored at -70°C. The plasma samples were analyzed using liquid chromatography tandem mass spectrometry (LC-MS / MS). The non-compartmental model analysis module of DAS 3.3.1 software was used to analyze the plasma concentration-time data of individual animals, and the pharmacokinetic parameters of the test compounds were calculated. The pharmacokinetic characteristics of the compounds in rats are shown in Table 16.

[0757] Table 16: Pharmacokinetic characteristics of the compounds

[0758]

[0759] As can be seen from Table 16, at the same single oral dose in rats, the systemic exposure of compound 2 is higher than that of control compound 4.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: Among them, R1 is selected from C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted with 0-6 substituents selected from F, Cl, and Br; R2 is selected from X is selected from CH, N, or O; n is 1; When X is O, it does not contain R a ; R3 is selected from C1-C6 alkyl; R4 is selected from C1-C6 alkyl and halogen; R6 is selected from hydrogen; R5 is selected from C1-C6 alkyl; R a selected from C1-C6 alkyl, amino and wherein the amino group is optionally substituted with 0-3 C1-C6 alkyl groups; Y is selected from N or O, where R' is C1-C6 alkyl; when Y is O, R' is absent; Z is selected from N.

2. The compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein R1 is selected from C1-C6 alkyl; the C1-C6 alkyl is optionally substituted with 0-3 F atoms; R2 is selected from R a selected from C1-C6 alkyl or R3 is selected from C1-C6 alkyl; R4 is selected from halogen and C1-C6 alkyl; R5 is selected from C1-C6 alkyl; R6 is hydrogen.

3. The compound or a pharmaceutically acceptable salt thereof according to claim 2, wherein R3 is selected from C1-C6 alkyl; R4 is selected from Cl, Br, and C1-C6 alkyl; R5 is C1-C6 alkyl; R6 is hydrogen.

4. The compound or a pharmaceutically acceptable salt thereof according to claim 3, wherein R2 is selected from R a selected from C1-C3 alkyl; wherein Y is selected from N or O, Z is selected from N; when Y is O, R' is absent; R3 is selected from C1-C6 alkyl; R4 is selected from Cl, Br, C1-C6 alkyl; R5 is selected from C1-C6 alkyl; R' is selected from C1-C6 alkyl.

5. The compound or a pharmaceutically acceptable salt thereof according to claim 4, wherein R1 is selected from methyl, ethyl, isopropyl, CF2H, CH2CF3; R2 is selected from R3 is selected from methyl, ethyl, and isopropyl; R4 is selected from Cl, Br, CH3; R5 is selected from methyl, ethyl, and isopropyl.

6. The compound or a pharmaceutically acceptable salt thereof according to claim 5, wherein R1 is selected from methyl, ethyl, isopropyl, -CF2H; R2 is selected from R3 is selected from methyl, ethyl, and isopropyl; R4 is selected from Br; R5 is selected from methyl, ethyl, and isopropyl.

7. A compound or a pharmaceutically acceptable salt thereof, selected from 8. A pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, and a pharmaceutically acceptable carrier.

9. A pharmaceutical composition comprising the compound according to any one of claims 1-7 and other anti-cancer drugs.

10. A pharmaceutical composition comprising the compound according to any one of claims 1-7 and other anti-tumor drugs.

11. The pharmaceutical composition according to claim 9, wherein the anti-cancer drug is one or more of cytotoxic drugs, hormonal drugs, antimetabolic drugs, tumor-targeted drugs, PARP inhibitor drugs, adjuvant therapy drugs, or anti-tumor biologics.

12. The pharmaceutical composition according to claim 10, wherein the anti-tumor drug is one or more of cytotoxic drugs, hormonal drugs, antimetabolic drugs, tumor-targeted drugs, PARP inhibitor drugs, adjuvant therapy drugs, or anti-tumor biologics.

13. The pharmaceutical composition according to claim 11 or 12, wherein the cytotoxic drug is one or more of carboplatin, cisplatin, irinotecan, paclitaxel, fluorouracil, cytarabine, lenalidomide, retinoic acid; the hormonal drug is one or more of dexamethasone, fulvestrant, tamoxifen; the antimetabolic drug is one or more of fluorouracil, methotrexate, tegafur, cytarabine; the tumor-targeted drug is one or more of imatinib, erlotinib, lapatinib; the PARP inhibitor drug is one or more of Olaparib, Rubraca, Zejula; the adjuvant therapy drug is one or more of recombinant human granulocyte colony-stimulating factor, erythropoietin, pamidronate disodium, zoledronic acid; the anti-tumor biologic drug is one or more of Keytruda, Opdiv, Tecentriq, Imfinzi, Bavencio.

14. A pharmaceutical preparation, comprising the compound according to any one of claims 1-7 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

15. Use of the compound according to any one of claims 1-7 or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to any one of claims 8-13, or the pharmaceutical preparation according to claim 14 in the preparation of an EGFR inhibitor or an ALK inhibitor or an EGFR and ALK inhibitor.

16. The use according to claim 15, wherein the EGFR inhibitor or ALK inhibitor or EGFR and ALK inhibitor is applied to plasmacytoma, mantle cell lymphoma, multiple myeloma, melanoma, breast cancer, liver cancer, cervical cancer, lung cancer, lymphoma, leukemia, ovarian cancer, kidney cancer, gastric cancer, nasopharyngeal cancer, thyroid cancer, pancreatic cancer, prostate cancer, adenocarcinoma, oral cancer, esophageal cancer, squamous cell carcinoma or colon cancer.

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