CN104341416B

CN104341416B - Protein tyrosine kinase inhibitor and its application

Info

Publication number: CN104341416B
Application number: CN201310329248.8A
Authority: CN
Inventors: 王勇; 赵立文; 陈宏雁; 王德忠; 刘阳; 毕胜; 高毅平; 张迪; 张仓
Original assignee: Nanjing Sanhome Pharmaceutical Co Ltd
Current assignee: Nanjing Sanhome Pharmaceutical Co Ltd
Priority date: 2013-07-31
Filing date: 2013-07-31
Publication date: 2017-03-29
Anticipated expiration: 2033-07-31
Also published as: CN104341416A; TWI546304B; WO2015014283A1; TW201504239A

Abstract

The invention belongs to chemical medicine, there is provided a class has the compound of protein tyrosine kinase inhibitory activity or its pharmaceutically acceptable salt, isomers, solvate, crystallization or prodrug.The present invention also provides the preparation method of the compounds of this invention, the pharmaceutical composition of the compound containing the present invention or its pharmaceutically acceptable salt, isomers, solvate, crystallization or prodrug, and the compound and composition of the present invention is for the application in the medicine for the treatment of or prevention of tumor.

Description

Protein tyrosine kinase inhibitor and its application

Technical field

The invention belongs to chemical medicine, and in particular to a class has the compound of protein tyrosine kinase inhibitory activity Or its pharmaceutically acceptable salt, isomers, solvate, crystallization or prodrug, and the pharmaceutical composition containing these compounds With the application of these compounds or composition in medicine preparation.

Background technology

Protein tyrosine kinase（PTKs）It is that a class can be catalyzed γ-phosphoric acid on ATP and be transferred on protein-tyrosine residue Kinases, by be catalyzed multiple protein tyrosine residue on phenolic hydroxyl group occur phosphorylation, and then activate functional protein effect Protein enzyme system.Protein tyrosine kinase（PTKs）In occupation of highly important status in signal transduction pathway in the cell, Adjust a series of physiological and biochemical procedures such as cell growth, differentiation, death.The unconventionality expression of protein tyrosine kinase can cause carefully Born of the same parents' propagation regulation gets muddled, and then leads oncogenic generation.Additionally, the unconventionality expression of protein tyrosine kinase also with tumour Invasion and attack and transfer, tumor neovasculature generation, the chemotherapy resistance to the action of a drug of tumour are closely related.Tyrosine kinase inhibitor can conduct Atriphos（ATP）The competitive inhibitor combined with EGFR-TK, competitive binding EGFR-TK block tyrosine The activity of kinases, suppresses cell propagation, has had several tyrosine protein kinase inhibitors to be successfully obtained exploitation.

Chronic myelocytic leukemia（CML）Chromosome translocation in patient forms BCR-ABL fusions, the BCR- of expression ABL protein tyrosine kinases are played an important role in cell signalling and conversion, are acted on by phosphorylation etc., are promoted The unlimited hyperplasia of CML maturation granulocytes.BCR-ABL is not expressed in normal cell, has become the ideal medicament for the treatment of CML Target.

Imatinib is the protein tyrosine kinase inhibitor of first molecular targeted therapy, Reverse transcriptase Adenosine triphosphate Glycosides (ATP) and thymidine kinase（TK）Acceptor（Such as KIT）Binding site, block TK phosphorylations, so as to suppress signal transduction, and can Suppress the KIT of the KIT mutation relevant with kinases and wild type, there is therapeutic effect to various types of cancers.Imatinib exists Inside and outside can suppress BCR-ABL EGFR-TKs on a cellular level, Selective depression BCR-ABL positive cell lines cell with And Philadelphia Chromosome Positive（Ph+）Chronic myelogenous leukemia（CML）With the fresh white blood of ALL patient The propagation of sick cell, induces its apoptosis.Additionally, Imatinib can also suppress platelet derived growth factor（PDGF）Acceptor The receptor tyrosine kinase c-Kit of EGFR-TK and stem cell factor (SCF), so as to suppress by PDGF and stem cell factor The cell behavior of mediation.Clinically it is mainly used in treating chronic myelocytic leukemia (CML) CML-BC, accelerated period or alpha-interferon Chronic phase patient after Endodontic failure, it is unable to surgery excision or the malignant gastrointestinal mesenchymal neoplasm of transfer occurs（GIST）Patient； It is also used for treating the positive patients with gastrointestinal stromal tumors (GIST) of CD117.

The exploitation of Imatinib and Clinical practice open the New Times of tumor cells targeting.But long-term taking she horse replace Buddhist nun, can produce drug resistance, cause disease relapse.With Imatinib extensive application clinically, resistance problems become increasingly conspicuous. The point mutation for being primarily due to BCR-ABL of acquired tolerance causes Imatinib be combined with BCR-ABL and be produced 's.And, it has been found that hundreds of BCR-ABL point mutation is related to imatinib-resistant, wherein 15～20% imatinib resistant There is T315I mutation in patient.

The appearance of imatinib-resistant, has evoked the research and development upsurge of tyrosine kinase inhibitor of new generation, to exploitation Go out more excellent for treating leukaemia, such as new medicine of each phase CML, Ph+ALL of resistance or non-resistance.

The content of the invention

It is an object of the invention to provide a class of formula I has the compound or its medicine of the BCR-ABL inhibitions of wide spectrum Acceptable salt, isomers, solvate, crystallization or prodrug are learned,

Wherein：

R₁Selected from hydrogen, alkyl, alkoxyl, haloalkyl, halogenated alkoxy ,-OH ,-NH₂, halogen and CN；

R₂Selected from hydrogen, alkyl, alkoxyl, haloalkyl, halogenated alkoxy ,-OH ,-NH₂, halogen and CN；

R₃Selected from hydrogen, alkyl, haloalkyl, halogen, aminoacyl, alkyl monosubstituted amino acyl group, double alkylaminoacyls and Cyano group；

X is selected from N and C (R₄), wherein R₄Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl acyl ammonia Base, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, Or R₄Connected carbon atom forms C (=O), described amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl Acylamino-, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl sulphonyl Amino can be replaced by one or more halogens, alkyl, amino, cyano group；

Y is selected from N, NH and C (R₅), wherein R₅Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl acyl Amino, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl sulphonyl ammonia Base, described amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroaryl acyl ammonia Base, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be by one or more halogens, alkane Base, amino, cyano group replace；

Singly-bound or double bond are represented, when X is C (=O),Represent singly-bound；

It is not following compound that condition is the compound：

3- ((2- amino-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- methyl-N- [4- ((4- first Base piperazine -1- bases) methyl) -3- trifluoromethyls] benzamide；

3- ((2- methylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- methyl-N- [4- ((4- Methylpiperazine-1-yl) methyl) -3- trifluoromethyls] benzamide；

3- ((2- dimethylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide.

It is a further object to provide the compounds of formula I or its pharmaceutically acceptable salt of the preparation present invention, The method of isomers, solvate, crystallization or prodrug.

It is also another object of the present invention to provide comprising the present invention compounds of formula I or its pharmaceutically acceptable salt, The composition of isomers, solvate, crystallization or prodrug and pharmaceutically acceptable carrier, and the formula I comprising the present invention Compound or its pharmaceutically acceptable salt, isomers, solvate, crystallization or prodrug and another kind of or multiple protein tyrosine-kinase The composition of enzyme inhibitor.

The present invention's a further object is the compounds of formula I or its pharmaceutically acceptable salt, isomery for providing the present invention The treatment of body, solvate, crystallization or prodrug and/or the method for prevention of tumor, and the compounds of formula I or its medicine of the present invention Learn acceptable salt, isomers, solvate, crystallization or prodrug prepare for treatment and/or prevention of tumor medicine in Using.

For above-mentioned purpose, the present invention provides technical scheme below：

In a first aspect, the present invention provides compounds of formula I or its pharmaceutically acceptable salt, isomers, solvate, knot Brilliant or prodrug,

Wherein：

It is not following compound that condition is the compound：

In the embodiment of some compound of Formula I, X is N, and Y is N,Represent double bond.

In the embodiment of some compound of Formula I, X is N, and Y is C (R₅),Represent double bond, R₅Selected from hydrogen, ammonia Base, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, Alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described amino, alkyl monosubstituted amino, double alkyl aminos, acyl It is amino, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, miscellaneous Arenesulfonyl amino can be replaced by one or more halogens, alkyl, amino, cyano group.

In the embodiment of some compound of Formula I, X is C (R₄), Y is N,Represent double bond, described R₄Choosing From hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulphur Acylamino-, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described amino, alkyl monosubstituted amino, double alkyl Amino, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, arylsulfonyl Amino, heteroaryl ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group.

In the embodiment of some compound of Formula I, X is C (=O), and Y is NH,Represent singly-bound.

In the embodiment of some compound of Formula I, X is C (R₄), Y is C (R₅),Double bond is represented, it is described R₄、R₅Separately selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, Heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described amino, list Alkyl amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulphur Acylamino-, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group.

In the embodiment of some preferred compound of Formula I, R₁Selected from hydrogen, C_1-6Alkyl, C_1-6Alkoxyl, halo C_1-6Alkyl, halo C_1-6Alkoxyl ,-OH ,-NH₂, halogen and CN；R₂Selected from hydrogen, C_1-6Alkyl, C_1-6Alkoxyl, halo C_1-6Alkane Base, halo C_1-6Alkoxyl ,-OH ,-NH₂, halogen and CN；R₃Selected from hydrogen, C_1-6Alkyl, halo C_1-6Alkyl, halogen, aminoacyl, Single C_1-6Alkylaminoacyl, double C_1-6Alkylaminoacyl and cyano group.

In the embodiment of some preferred compound of Formula I, R₁Selected from hydrogen, C_1-4Alkyl, C_1-4Alkoxyl, halo C_1-4Alkyl, halo C_1-4Alkoxyl ,-OH ,-NH₂, halogen and CN；R₂Selected from hydrogen, C_1-4Alkyl, C_1-4Alkoxyl, halo C_1-4Alkane Base, halo C_1-4Alkoxyl ,-OH ,-NH₂, halogen and CN；R₃Selected from hydrogen, C_1-4Alkyl, halo C_1-4Alkyl, halogen, aminoacyl, Single C_1-4Alkylaminoacyl, double C_1-4Alkylaminoacyl and cyano group.

It is highly preferred that in the embodiment of compound of Formula I, R₁Selected from hydrogen, C_1-3Alkyl, C_1-3Alkoxyl, halo C_1-3 Alkyl, halo C_1-3Alkoxyl ,-OH ,-NH₂, halogen and CN；R₂Selected from hydrogen, C_1-3Alkyl, C_1-3Alkoxyl, halo C_1-3Alkyl, halogen For C_1-3Alkoxyl ,-OH ,-NH₂, halogen and CN；R₃Selected from hydrogen, C_1-3Alkyl, halo C_1-3Alkyl, halogen, aminoacyl, list C_1-3 Alkylaminoacyl, double C_1-3Alkylaminoacyl and cyano group.

Preferably, the present invention provides the compound or its pharmaceutically acceptable salt, isomers, solvate, knot of formula Ia Brilliant or prodrug,

Wherein：

Y is selected from N, NH and C (R₅), wherein R₅Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl acyl Amino, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl sulphonyl ammonia Base, described amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroaryl acyl ammonia Base, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be by one or more halogens, alkane Base, amino, cyano group replace；Singly-bound or double bond are represented, when X is C (=O),Represent singly-bound；

When X is N, Y is C (R₅), and the acetenyl are connected by the pyridine ring that condensed with which of the ring containing X, Y and pyridine ring Together constitute [1,2,4]-triazole simultaneously [1,5-a] pyridine -7- ethynyl groups when, R₅Selected from hydrogen, acylamino-, alkyl acyl ammonia Base, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, Described acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, aryl sulphur Acylamino-, heteroaryl ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group.

In the embodiment of some formulas Ia compounds, X is N, and Y is N,Show double bond.

In the embodiment of some formulas Ia compounds, X is N, and Y is C (R₅),Double bond is represented, and contains X, Y The pyridine ring that condensed with which of ring and the acetenyl that is connected of pyridine ring together with constitute [1,2,4]-triazole simultaneously [1,5-a] pyrrole During pyridine -7- ethynyl groups, R₅Selected from hydrogen, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulphonyl ammonia Base, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described acylamino-, alkyl amido, aryl acyl ammonia Base, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino can by one or Multiple halogens, alkyl, amino, cyano group replace.

In the embodiment of some formulas Ia compounds, X is N, and Y is C (R₅),Double bond is represented, and contains X, Y The pyridine ring that condensed with which of ring and the acetenyl that is connected of pyridine ring together with constitute [1,2,4]-triazole simultaneously [1,5-a] pyrrole During pyridine -6- ethynyl groups, R₅Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, aryl Acylamino-, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, it is described Amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulphonyl ammonia Base, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be by one or more halogens, alkyl, amino, cyanogen Base replaces.

In the embodiment of some formulas Ia compounds, X is C (R₄), Y is N,Represent double bond, described R₄Choosing From hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulphur Acylamino-, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described amino, alkyl monosubstituted amino, double alkyl Amino, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, arylsulfonyl Amino, heteroaryl ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group.

In the embodiment of some formulas Ia compounds, X is C (=O), and Y is NH,Represent singly-bound.

In the embodiment of some formulas Ia compounds, X is C (R₄), Y is C (R₅),Double bond is represented, it is described R₄、R₅Separately selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, Heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described amino, list Alkyl amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulphur Acylamino-, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group.

In the embodiment of some preferred formula Ia compounds, R₁Selected from hydrogen, C_1-6Alkyl, C_1-6Alkoxyl, halo C_1-6Alkyl, halo C_1-6Alkoxyl ,-OH ,-NH₂, halogen and CN；R₂Selected from hydrogen, C_1-6Alkyl, C_1-6Alkoxyl, halo C_1-6Alkane Base, halo C_1-6Alkoxyl ,-OH ,-NH₂, halogen and CN；R₃Selected from hydrogen, C_1-6Alkyl, halo C_1-6Alkyl, halogen, aminoacyl, Single C_1-6Alkylaminoacyl, double C_1-6Alkylaminoacyl and cyano group.

In the embodiment of some preferred formula Ia compounds, R₁Selected from hydrogen, C_1-3Alkyl, C_1-3Alkoxyl, halo C_1-3Alkyl, halo C_1-3Alkoxyl ,-OH ,-NH₂, halogen and CN；R₂Selected from hydrogen, C_1-3Alkyl, C_1-3Alkoxyl, halo C_1-3Alkane Base, halo C_1-3Alkoxyl ,-OH ,-NH₂, halogen and CN；R₃Selected from hydrogen, C_1-3Alkyl, halo C_1-3Alkyl, halogen, aminoacyl, Single C_1-3Alkylaminoacyl, double C_1-3Alkylaminoacyl and cyano group.

It is further preferred that the present invention provides the compound or its pharmaceutically acceptable salt, isomers, solvent of formula I-a Compound, crystallization or prodrug,

Wherein：

R₅Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroaryl Base acylamino-, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described amino, monoalkyl Amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkylsulfonylamino group Base, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group；

It is not following compound that condition is the compound：

It is further preferred that the compound or its pharmaceutically acceptable salt, isomers, solvent of this offer formula I-b are closed Thing, crystallization or prodrug,

Wherein：

Y is selected from N and NH；

R₄Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroaryl Base acylamino-, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, or R₄With the carbon being connected Atom forms C (=O), described amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, Heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be by one or many Individual halogen, alkyl, amino, cyano group replace；

Singly-bound or double bond are represented, works as R₄When connected carbon atom forms C (O),Singly-bound is represented, Y is NH。

It is further preferred that the present invention provides the compound or its pharmaceutically acceptable salt, isomers, solvent of formula Ia-a Compound, crystallization or prodrug,

Wherein：

R₅Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroaryl Base acylamino-, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described amino, monoalkyl Amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkylsulfonylamino group Base, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group.

When [1,2,4]-triazole constitute together with the be connected acetenyl of pyridine ring and pyridine ring for being condensed [1,2,4]- Triazole simultaneously [1,5-a] pyridine -7- ethynyl groups when, described R₅Selected from hydrogen, acylamino-, alkyl amido, aryl acyl ammonia Base, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described acyl ammonia Base, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl Ylsulfonylamino can be replaced by one or more halogens, alkyl, amino, cyano group.

It is further preferred that the present invention provides the compound or its pharmaceutically acceptable salt of formula I-a ', isomers, molten Agent compound, crystallization or prodrug,

Wherein：

It is further preferred that the present invention provides formula I-a " compound or its pharmaceutically acceptable salt, isomers, molten Agent compound, crystallization or prodrug,

Wherein：

R₅Selected from hydrogen, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroaryl acyl Amino, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described acylamino-, alkyl acyl ammonia Base, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino Can be replaced by one or more halogens, alkyl, amino, cyano group；

It is not following compound that condition is the compound：

3- ((2- dimethylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide..

It is further preferred that the present invention provides the compound or its pharmaceutically acceptable salt of formula I-b ', isomers, molten Agent compound, crystallization or prodrug,

Wherein：

Y is selected from N and NH；

R₄Selected from hydrogen, amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, acrylamido, heteroaryl Base acylamino-, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, or R₄It is connected with which Carbon atom forms C (=O), described amino, alkyl monosubstituted amino, double alkyl aminos, acylamino-, alkyl amido, aryl acyl ammonia Base, heteroarylamido, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino can by one or Multiple halogens, alkyl, amino, cyano group replace；

Singly-bound or double bond are represented, works as R₄When the carbon atom being connected with which forms C (=O),Represent singly-bound, Y is NH.

It is further preferred that the present invention provides formula I-b " compound or its pharmaceutically acceptable salt, isomers, molten Agent compound, crystallization or prodrug,

Wherein：

Y is selected from N and NH；

It is further preferred that the present invention provide formula Ia-a ' compound or its pharmaceutically acceptable salt, isomers, Solvate, crystallization or prodrug,

Wherein：

R₅Selected from hydrogen, acylamino-, alkyl amido, acrylamido, heteroarylamido, sulfonamido, alkyl sulfonyl Amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino, described acylamino-, alkyl amido, acrylamido, heteroaryl acyl Amino, sulfonamido, alkyl sulfonyl amino, Arenesulfonyl amino, heteroaryl ylsulfonylamino can be by one or more halogens, alkane Base, amino, cyano group replace.

In a preferred embodiment of the present invention, R₁Selected from hydrogen, alkyl, haloalkyl, halogen and CN.

In further preferred embodiment of the present invention, R₁Selected from methyl and trifluoromethyl.

In a preferred embodiment of the present invention, R₂Selected from hydrogen, alkyl, haloalkyl, halogen and CN.

In further preferred embodiment of the present invention, R₂Selected from methyl, ethyl, propyl group and isopropyl.

In a preferred embodiment of the present invention, R₃Selected from hydrogen, C_1-6Alkyl, halo C_1-6Alkyl, halogen, aminoacyl, Single C_1-6Alkylaminoacyl, double C_1-6Alkylaminoacyl and cyano group.

In further preferred embodiment of the present invention, R₃Selected from hydrogen, methyl, ethyl, propyl group, isopropyl, trifluoromethyl, three Fluoro ethyl, halogen and methylamino acyl group.

In a preferred embodiment of the present invention, R₄Selected from amino, list C_1-6Alkyl amino, double C_1-6Alkyl amino, formyl Amino, acetylamino, propionamido, benzoyl amino, trifluoroacetamido, methanesulfonamido, second sulfonamido, benzene fulfonic amide Base, or R₄C (=O) is formed with the carbon atom being connected.

In further preferred embodiment of the present invention, R₄Selected from amino, methylamino, ethylamino, the third amino, isopropylamino, Dimethylamino, lignocaine, dipropyl amino, methylethylamine, methylpropylamino, ethylpropylamino, formamido group, second Acylamino-, propionamido, trifluoroacetamido, methanesulfonamido, second sulfonamido, or R₄Formed with the carbon atom being connected C(=O)。

In a preferred embodiment of the present invention, R₅Selected from amino, list C_1-6Alkyl amino, double C_1-6Alkyl amino, formyl Amino, acetylamino, propionamido, benzoyl amino, trifluoroacetamido, methanesulfonamido, second sulfonamido, benzene fulfonic amide Base.

In further preferred embodiment of the present invention, R₅Selected from amino, methylamino, ethylamino, the third amino, isopropylamino, Dimethylamino, lignocaine, dipropyl amino, methylethylamine, methylpropylamino, ethylpropylamino, formamido group, second Acylamino-, propionamido, trifluoroacetamido, methanesulfonamido, second sulfonamido.

The invention provides compound in detail below：

On the other hand, the present invention provides the preparation method of the general formula compound of the present invention.The preparation side of compounds of formula I Method comprises the steps：

(1) preparation of the intermediate of formula 5：

A) compound of formula 1 reacts the intermediate for obtaining formula 3 with the compound of formula 2；

B) intermediate of formula 3 reacts the intermediate for obtaining formula 4 with trimethylsilyl acetylene；

C) intermediate of formula 4 sloughs the intermediate that TMS obtains formula 5；

(2) preparation of compounds of formula I：

D) intermediate of formula 5 obtains compounds of formula I with the intermediate reaction of formula 6.

Wherein, R₁、R₂、R₃、R₅, X, Y there is implication in formula I, M is selected from chlorine, bromine, iodine.

Similarly, those skilled in the art can refer to the preparation method of compounds of formula I of the present invention, prepare the present invention logical Formulas I, formula Ia, formula I-a, formula I-b, formula Ia-a, formula I-a ', formula I-a ", formula I-b ', formula I-b " and formula The compound of Ia-a '.

Specifically, the preparation method of the compound of formula I-a comprises the steps：

（1）The preparation of the intermediate of formula 9：

E) compound of formula 7 reacts the intermediate for obtaining formula 8 with different sulphur cyanato- Ethyl formate；

F) intermediate of formula 8 reacts the intermediate for obtaining formula 9 with hydroxylamine hydrochloride；

（2）The synthesis of the intermediate of formula 10

G) intermediate of formula 9 is obtained formula 10 through conventional alkylated reaction, acylation reaction or sulfonylating reaction Intermediate；

（3）The preparation of the compound of formula I-a：

Or

H) intermediate of the intermediate of formula 9 or formula 10 obtains formula I-a compounds with the intermediate reaction of formula 5；

Wherein, R₁、R₂、R₃And R₅With the implication in formula I-a, M is selected from chlorine, bromine, iodine.

Specifically, wherein R₄Connected carbon atom forms C (=O), preparation sides of the Y for the compound of the formula I-b of NH Method comprises the steps：

（1）The preparation of the intermediate of formula 13

I) intermediate of formula 11 and hydration hydrazine reaction obtain the intermediate of formula 12；

J) intermediate of formula 12 reacts the intermediate for obtaining formula 13 with triphosgene；

（2）The preparation of the target compound of Formulas I-b

K) intermediate of formula 13 obtains wherein R with the intermediate reaction of formula 5₄Connected carbon atom forms C (=O), Y For the compound of the formula I-b of NH；

Wherein, R₁、R₂、R₃With the implication in formula I-b, M is selected from chlorine, bromine, iodine.

The third aspect, the present invention provide pharmaceutical composition, its include the present invention compound or its pharmaceutically acceptable salt, Isomers, solvate, crystallization or prodrug.

In some embodiments, the present invention provides pharmaceutical composition, and which includes the compound of the present invention, isomers, molten Agent compound, crystallization or prodrug, also comprising one or more selected from following composition：Tyrosine protein enzyme inhibitor, EGFR suppress Agent, VEGFR inhibitor, BCR-ABL inhibitor, c-kit inhibitor, c-Met inhibitor, Raf inhibitor, mek inhibitor, group egg White deacetylase inhibitor, VEGF antibody, EGF antibody, HIV kinases inhibitors, HMG-CoA reductase inhibitor etc..

Can by the compound of the present invention, isomers, solvate, crystallization or prodrug and pharmaceutically acceptable carrier, Diluent or excipient are prepared by mixing into pharmaceutical preparation, to be suitable for oral or parenteral.Medication includes, but does not limit In intracutaneous, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal and peroral route.The preparation can be applied by any approach, example Such as by being transfused or injecting, applied by the approach that transepithelial or mucocutaneous (such as oral mucosa or rectum etc.) absorb.Give Medicine can be whole body or local.The example of oral administration preparation includes solid or liquid dosage form, specifically, including piece Agent, pill, granula, pulvis, capsule, syrup, emulsion, supensoid agent etc..The preparation can pass through methods known in the art system It is standby, and include the conventional use of carrier of field of pharmaceutical preparations, diluent or excipient.

Fourth aspect, the present invention provide compound, isomers, solvate, crystallization or prodrug or the present invention of the present invention Medicine composite for curing or prevention of tumor method and prepare prevention or tumor in application, including to tumour Easily send out crowd or tumor patient applies compound, isomers, solvate, crystallization or the prodrug of the present invention or comprising the present invention Compound, isomers, solvate, the pharmaceutical composition of crystallization or prodrug, effectively to reduce Tumor incidence, extend tumour Patient vitals.

Term explanation

" alkyl " of the present invention refers to the saturated hydrocarbyl of straight or branched, preferably C_1-6Alkyl, more preferably C_1-3 Alkyl, suitable C_1-3Alkyl is methyl, ethyl, propyl group, isopropyl.

" alkoxyl " of the present invention refers to alkyl-O-, preferably C_1-6Alkyl-O-, more preferably C_1-3Alkyl-O-, Suitable C_1-3Alkyl-O- is methoxyl group, ethyoxyl, propoxyl group, isopropoxy.

" halogen " of the present invention refers to fluorine, chlorine, bromine, iodine, preferably fluorine, chlorine.

" haloalkyl " of the present invention is referred at least by the alkyl of a halogen substiuted, preferably halo C_1-6Alkyl, enters one Step is preferably halo C_1-3Alkyl, suitable halo C_1-3Alkyl is chloromethyl, methyl fluoride, dichloromethyl, difluoromethyl, three chloromethanes Base, trifluoromethyl, chloroethyl, fluoro ethyl, Dichloroethyl, two fluoro ethyls, trichloroethyl, trifluoroethyl.

" halogenated alkoxy " of the present invention is referred at least by the alkoxyl of a halogen substiuted, preferably at least by a halogen The C that element replaces_1-6Alkoxyl, more preferably halo C_1-3Alkoxyl, suitable halo C_1-3Alkoxyl is chloromethane epoxide, fluorine Methoxyl group, dichloro methoxyl group, difluoro-methoxy, trichloromethoxy, trifluoromethoxy；Two chloroethoxies, difluoroethoxy, trichlorine Ethyoxyl, trifluoro ethoxy.

" acylamino- " of the present invention refers to HC (O)-NH-.

" alkyl amido " of the present invention refers to alkyl-C (O)-NH-, preferably C_1-6Alkyl-C (O)-NH-, it is further excellent Elect C as_1-3Alkyl-C (O)-NH-.

" acrylamido " of the present invention refers to aryl-C (O)-NH-.

" heteroarylamido " of the present invention refers to heteroaryl-C (O)-NH-.

" sulfonamido " of the present invention refers to HS (O)₂-NH-。

" the alkyl sulfonyl amino " of the present invention refers to alkyl-S (O)₂- NH-, preferably C_1-6Alkyl-S (O)₂- NH-, enters one Step is preferably C_1-3Alkyl-S (O)₂-NH-。

" Arenesulfonyl amino " of the present invention refers to aryl-S (O)₂-NH-。

" the heteroaryl ylsulfonylamino " of the present invention refers to heteroaryl-S (O)₂-NH-。

" aryl " of the present invention refers to the aromatic hydrocarbon group containing one or more phenyl ring.Suitable aryl includes phenyl, naphthalene Base.

" heteroaryl " of the present invention refers to the aromatic radical that at least one carbon atom is substituted by hetero atom in aryl.Institute The hetero atom stated is O, S, N.

" solvate " of the present invention refers to solute in a conventional sense（Such as reactive compound, the salt of reactive compound）With Solvent（Such as water）The compound that combination is formed.Solvent refers to the solvent of known to those of skill in the art or easy determination.Such as Fruit is water, then solvate is commonly referred to as hydrate, such as monohydrate, dihydrate, trihydrate etc..

" crystallization " of the present invention refers to the various solid forms that compound of the present invention is formed, including crystal formation, nothing are determined Shape.

" isomers " of the present invention refers to the isomers of the cis or trans configuration of compound.Therefore, the present invention includes often Plant cis-isomer or transisomer and be substantially free of the mixture of other isomers and these isomers.

" prodrug " of the present invention refer under the physiological condition of organism, due to the reaction such as enzyme, hydrochloric acid in gastric juice and conversion cost The compound of the compound of invention, i.e., the compound of the compound invented by conversion costs such as the oxidation of enzyme, reduction, hydrolysis And/or the compound of the compound of the conversion cost such as the hydrolysis by hydrochloric acid in gastric juice etc. invention.

" the pharmaceutically acceptable salt " of the present invention refers to the pharmaceutically acceptable salt that the compound of the present invention is formed with acid, Described acid include but is not limited to phosphoric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, citric acid, maleic acid, malonic acid, mandelic acid, butanedioic acid, Fumaric acid, acetic acid, lactic acid, nitric acid etc..

The present invention " pharmaceutical composition " refer to comprising any compound as herein described, including isomers, prodrug, Solvate, pharmaceutically acceptable salt or its chemical forms of protection, and one or more pharmaceutically acceptable carrier is mixed Compound.

The present invention " pharmaceutically acceptable carrier " refer to organism is not caused obvious irritation and do not disturb to Give the carrier of the biologically active and property of compound, comprising solvent, diluent or other excipient, dispersant, surfactant, Isotonic agent, thickener or emulsifying agent, preservative, solid binder, lubricant etc..Unless any conventional carrier medium and the present invention Compound is incompatible.Carbohydrate, such as lactose, Portugal can be included, but are not limited to as some examples of pharmaceutically acceptable carrier Grape sugar and sucrose；Starch, such as cornstarch and farina；Cellulose and its derivates, such as sodium carboxymethylcellulose and Cellulose and cellulose acetate；Malt, gelatin etc..

" excipient " of the present invention refers to and is added in Pharmaceutical composition further to promote to give the inert substance of compound. Excipient can include calcium carbonate, calcium phosphate, various saccharides and polytype starch, cellulose derivative, gelatin, plant Oil, polyethylene glycol.

" application in the medicine for treatment or prevention of tumor is prepared " of the present invention refers to the life that can suppress tumour Long, development and/or transfer, mainly give the compound for controlling the present invention for giving treatment effective dose to press down to required human or animal System, the growth for slowing down or reversing subject's tumour, development or expanding.

The compound of the present invention refers to all of general formula compound of the invention, including formula I, formula Ia, formula I-a, logical Change described in Formulas I-b, formula Ia-a, formula I-a ', formula I-a ", formula I-b ', formula I-b " and the arbitrary formulas of formula Ia-a ' Compound and particular compound.

Specific embodiment

Representational embodiment is in order to the present invention is better described, not for the protection model for limiting the present invention below Enclose.

Embodiment 13- ((2- methylamino -7- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- Methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of step 11- (4- methyl -5- bromopyridine -2- bases) -3- ethoxycarbonylthioureas

In 50mL round-bottomed flasks, under condition of ice bath, addition 2- amino -4- methyl -5- bromopyridines (1.87g, 10mmol), dichloromethane (DCM, 20mL) is added, different sulphur cyanato- Ethyl formate is added（EtOC (O) NCS, 1.4g, 10.6mmol), 5 DEG C of reaction 15min, under normal temperature, then react 2h.40 DEG C of concentrations, add 50ml petroleum ethers（PE）, have substantial amounts of solid Body is separated out, and is filtered, and filter cake PE is washed 3 times, is dried, is obtained title compound.

ESI-MS m/z:[M+H]⁺=317.9。

The preparation of step 22- amino -6- bromine-7-methyls-[1,2,4] triazole simultaneously [1,5-a] pyridine

In 100mL eggplant-shape bottles, addition 1- (4- methyl -5- bromopyridine -2- bases) -3- ethoxycarbonylthioureas (1.21mg, 4mmol), hydroxylamine hydrochloride (1.38g, 20mmol) and DIPEA（DIEA, 1.65mL), add 40ml methyl alcohol and The mixed solution of ethanol（V:V=1:1）, 1h is stirred at room temperature, 70 DEG C of reaction 4h are warming up to.After reaction 4h, stop heating, be spin-dried for, plus Enter 20ml frozen water, filter, be dried, obtain title compound.

1H NMR(300MHz,DMSO-d6)δ:8.88(s,1H),7.36(s,1H),6.04(s,2H),2.37(s,3H)。

ESI-MS m/z:[M+H]⁺=227.1。

The preparation of -6 bromine-7-methyl of step 32- methylamino-[1,2,4] triazole simultaneously [1,5-a] pyridine

In dry two mouthfuls of flasks of 100ml, compound 2- amino -6- bromine-7-methyls-[1,2,4] triazole is added simultaneously [1,5-a] pyridine (200mg, 0.88mmol), paraformaldehyde (264mg, 8.8mmol), sodium methoxide（190mg, 3.52mmol）With Methyl alcohol（5ml）, nitrogen protection it is lower 80 DEG C reaction 2h, be cooled to room temperature, by sodium borohydride（167.2mg,4.4mmol）It is in batches careful It is added in reaction system, is heated to after 80 DEG C of stirring 2h under condition of ice bath, adds acetone that excessive sodium borohydride is quenched, it is dense Contracting.Add water（20ml）, ethyl acetate（EA, 50ml）Extraction three times, merges organic phase concentration, is dried, filter, column chromatography purifying Obtain title compound.

ESI-MS m/z:[M+H]⁺=241.1。

Step 4：The iodo- 4- methyl-N- of 3- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide Preparation

4- (4- methylpiperazine-1-yl methyl) -3- 5-trifluoromethylanilines are added in the reactor（2.27g, 8.3mmol）、 The iodo- 4- methyl benzoyl chlorides of 3-（10mmol）, 15ml tetrahydrofurans, 10ml triethylamines, be stirred at room temperature 4 hours.Reaction terminates, and uses Saturation NaHCO₃Solution is washed, ethyl acetate/water extraction, the washing of saturation NaCl solution, anhydrous Na₂SO₄It is dried, vacuum distillation is removed Solvent, silica column purification is gone to obtain title compound.

¹H NMR(500MHz,CDCl₃)δ:8.39(s,1H,N-H),8.29(s,1H,Ar-H),7.88(d,1H,Ar-H), 7.86(s,1H,Ar-H),7.75(d,1H,Ar-H),7.73(d,1H,Ar-H),7.28(d,1H,Ar-H),3.62(s,2H, PhCH₂),2.60(b,8H,4×-CH₂),2.47(s,3H,-CH₃),2.31(s,3H,-CH₃)。

Step 5：3- Trimethylsilanylethynyl -4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoros Aminomethyl phenyl] benzamide preparation

By 3- iodo- 4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide （3.1g, 6.1mmol）、Pd(PPh₃)₂Cl₂（426mg, 0.61mmol）、CuI（231mg, 1.21mmol）It is placed in reactor, plus Enter toluene 30ml and make solvent, triethylamine 1ml maintains alkaline environment.Under inert gas shielding, trimethyl is added in the mixture Silane ethyl-acetylene（3.0g, 30.3mmol）, 58 DEG C are stirred 24 hours.Reaction terminates, and in reactant mixture adds ethyl acetate Extracted with water, merge organic layer, washed with saturation NaCl solution, added anhydrous Na₂SO₄It is dried.Reduced pressure concentration, residue Jing silica column purifications obtain title compound.

¹H NMR(500MHz,CDCl₃)δ:8.30(s,1H,N-H),7.86(s,1H,Ar-H),7.83(d,1H,Ar-H), 7.72(s,1H,Ar-H),7.55(d,1H,Ar-H),7.41(d,1H,Ar-H),7.24(d,1H,Ar-H),3.60(s,2H, PhCH₂),2.48(b,8H,4×-CH₂),2.45(s,3H,-CH₃),2.28(s,3H,-CH₃),0.26(s,9H,3×-CH₃)。

Step 6：3- acetenyl -4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzene The preparation of formamide

Step 5 is reacted into gains (1.59g, 3.3mmol), potassium carbonate (1.82g, 13.2mmol), the mixing of 20ml methyl alcohol In reactor, under inert gas shielding, 3 hours are stirred at room temperature.Reaction terminates, and methyl alcohol is removed on Rotary Evaporators, adds second Acetoacetic ester and water are extracted, and are merged organic layer, are washed with saturation NaCl solution, add anhydrous Na₂SO₄It is dried.Then this is had Machine solution is concentrated on a rotary evaporator, and residue Jing silica column purifications obtain title compound.

¹H NMR(500MHz,CDCl₃)δ:10.47(s,1H,N-H),8.19(s,1H,Ar-H),8.08(s,1H,Ar-H), 8.04(d,1H,Ar-H),7.91(d,1H,Ar-H),7.70(d,1H,Ar-H),7.47(d,1H,Ar-H),4.50(s,1H,≡ CH),3.56(s,2H,PhCH₂),2.50(s,3H,-CH₃),2.36(b,8H,-4×CH₂),2.15(s,3H,-CH₃)。

Step 73- ((2- methylamino -7- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

Weigh 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzoyl Amine（126mg, 0.3mmol）, 2- methylamino -6- bromine-7-methyls-[1,2,4] triazole simultaneously [1,5-a] pyridine（60mg, 0.3mmol）, Pd (Pph₃)₂Cl₂（22mg, 0.03mmol）, three hexamethylene phosphines（Pcy3,16mg, 0.06mmol）, CuI（6mg, 0.03mmol）, cesium carbonate（100mg, 0.3mmol）, in adding tube sealing.Then in tube sealing 6 are added to drip diisopropyl ethyl amine （DIPEA）And 6mL DMF.Logical argon gas deoxygenation was sealed after 5 minutes, and 80 DEG C are stirred overnight.During each reactant adds 10mL tube sealings, Plus DMF is to half volume, lead to argon gas deoxygenation 5 minutes, sealing, 80 DEG C are stirred overnight.Next day reactant liquor ammonia scrubbing 2 times, second Acetoacetic ester is extracted 3 times, uses saturated common salt water washing several times after merging organic layer, adds anhydrous sodium sulfate drying, purifying to obtain Target compound.

1H NMR(300MHz,DMSO-d6)δ:10.54(s,1H),8.92(s,1H),8.20(s,1H),8.14(s,1H), 8.05(d,1H),7.92(d,1H),7.70(d,1H),7.52(d,1H),7.34(s,1H),6.58(q,1H),3.57(s,2H), 2.82(s,3H),2.57(s,3H),2.53(s,3H),2.38(m,8H),2.16(s,3H)。

ESI-MS m/z:[M+H]⁺=576.3。

Embodiment 23- ((2- amino -5- trifluoromethyls-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- amino -5- trifluoromethyl -6- simultaneously [1,5-a] pyridine

With the bromo- 6- trifluoromethyl pyridines of 2- amino -5- as raw material, title compound is prepared by 1 step 1 of embodiment and step 2 Thing.

1H NMR(300MHz,DMSO-d6)δ:7.77(d,1H),7.59(d,1H),6.48(s,2H)。

Step 23- ((2- amino -5- trifluoromethyls-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- The preparation of methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- amino -5- trifluoromethyl -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl - N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the side of 1 step 7 of embodiment Legal system obtains target compound.

1H NMR(300MHz,DMSO-d6)δ:10.52(s,1H),8.20(s,1H),8.16(s,1H),8.07(d,1H), 8.04(d,1H),7.76(d,1H),7.74(d,1H),7.71(d,1H),7.54(d,1H),6.61(s,2H),3.57(s,2H), 2.73(s,3H),2.40(m,8H),2.18(s,3H)。

ESI-MS m/z:[M+H]⁺=616.4。

Embodiment 33- ((2- amino -7- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of step 12- amino -6- bromine-7-methyls-[1,2,4] triazole simultaneously [1,5-a] pyridine

With 2- amino -4- methyl -5- bromopyridines as raw material, title compound is obtained by 1 step 1 of embodiment and step 2.

1H NMR(300MHz,DMSO-d6)δ:8.88(s,1H),7.36(s,1H),6.04(s,2H),2.37(s,3H)。

ESI-MS m/z:[M+H]⁺=227.1。

Step 23- ((2- amino -7- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With 2- amino -6- bromine-7-methyls-[1,2,4] triazole simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepared target compound.

1H NMR(300MHz,DMSO-d6)δ:10.54(s,1H),8.86(s,1H),8.21(s,1H),8.15(s,1H), 8.06(d,1H),7.92(d,1H),7.71(d,1H),7.52(d,1H),7.33(s,1H),6.13(s,2H),3.58(s,2H), 2.58(s,6H),2.40(m,8H),2.19(s,3H)。

ESI-MS m/z:[M+H]⁺=562.4。

Embodiment 43- ((2- amino -8- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of the bromo- 8- methyl of step 12- amino -6--[1,2,4] triazole simultaneously [1,5-a] pyridine

With 2- amino -3- methyl -5- bromopyridines as raw material, title compound is obtained by 1 step 1 of embodiment and step 2.

Step 23- ((2- amino -8- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With the bromo- 8- methyl of 2- amino -6--[1,2,4] triazole simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

¹H NMR(300MHz,DMSO-d₆)δ:10.52(s,1H),8.76(s,1H),8.21(s,1H),8.15(s,1H), 8.06(d,1H),7.91(d,1H),7.70(d,1H),7.51(d,1H),7.42(s,1H),6.16(s,2H),3.58(s,2H), 3.26(s,6H),2.42(m,8H),2.20(s,3H)。

ESI-MS m/z:[M+H]⁺=562.3。

Embodiment 53- ((2- amino -5- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- amino -5- methyl -6- simultaneously [1,5-a] pyridine

With 2- amino -6- methyl -5- bromopyridines as raw material, title compound is obtained by 1 step 1 of embodiment and step 2.

Step 23- ((2- amino -5- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- amino -5- methyl -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.52(s,1H),8.21(s,1H),8.17(s,1H),8.06(d,1H), 7.91(d,1H),7.71(d,1H),7.58(d,1H),7.52(s,1H),7.29(s,1H),6.21(s,2H),3.58(s,2H), 2.85(s,3H),2.58(s,3H),2.40(m,8H),2.19(s,3H)。

ESI-MS m/z:[M+H]⁺=562.5。

Embodiment 63- ((2- dimethylamino -5- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With 2- amino -6- methyl -5- bromopyridines as raw material, it is obtained by the method for 1 step 1 of embodiment and step 2 titled Compound.

The preparation of bromo- [1,2,4] triazoles of step 22- dimethylamino -5- methyl -6- simultaneously [1,5-a] pyridine

In 50mL eggplant-shape bottles, under condition of ice bath, add bromo- [1,2, the 4] triazoles of 2- amino -5- methyl -6- simultaneously [1, 5-a] pyridine (1.06mg, 5mmol), be dried THF20ml, add sodium hydride（240mg, 10mmol）, 15min is stirred, then Add 2ml iodomethane, normal-temperature reaction 4h.It is spin-dried for, adds 50ml water, ethyl acetate extraction（30ml*3）, column chromatography purifying obtain Title compound.

Step 33- ((2- dimethylamino -5- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- The preparation of methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- dimethylamino -5- methyl -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl - N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the side of 1 step 7 of embodiment Method prepares target compound.

1H NMR(300MHz,DMSO-d6)δ:10.53(s,1H),8.21(s,1H),8.17(s,1H),8.06(d,1H), 7.92(d,1H),7.71(d,1H),7.64(d,1H),7.53(d,1H),7.37(s,1H),3.57(s,2H),3.31(s,3H), 3.08(s,3H),2.88(s,3H),2.58(s,3H),2.39(m,8H),2.18(s,3H)。

ESI-MS m/z:[M+H]⁺=590.3。

Embodiment 73- ((2- dimethylamino -8- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- dimethylamino -8- methyl -6- simultaneously [1,5-a] pyridine

With 2- amino -3- methyl -5- bromopyridines as raw material, title is prepared according to the method for 6 step 1 of embodiment and step 2 Compound.Step 23- ((2- dimethylamino -8- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- The preparation of methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With -6 bromo- [1,2,4] triazole of 2- dimethylamino -8- methyl simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl - N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the side of 1 step 7 of embodiment Method prepares target compound.

1H NMR(300MHz,DMSO-d6)δ:10.52(s,1H),8.87(s,1H),8.22(s,1H),8.16(s,1H), 8.07(d,1H),7.92(d,1H),7.72(d,1H),7.53(d,1H),7.48(s,1H),3.58(s,2H),3.31(s,3H), 3.08(s,3H),2.88(s,3H),2.58(s,3H),2.37(m,8H),2.18(s,3H)。

ESI-MS m/z:[M+H]⁺=590.3。

Embodiment 83- ((2- methylamino -8- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- Methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- methylamino -8- methyl -6- simultaneously [1,5-a] pyridine

With 2- amino -3- methyl -5- bromopyridines as raw material, title compound is prepared according to the method for 1 step 1-3 of embodiment Thing.

¹H NMR(300MHz,DMSO-d₆)δ:8.83-8.79(m,1H),7.43(s,1H),6.53-6.52(m,1H), 3.32(s,3H),2.39(s,3H)。

ESI-MS m/z:[M+H]⁺=241.0。

Step 23- ((2- methylamino -8- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- methylamino -8- methyl -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.52(s,1H),8.82(s,1H),8.21(s,1H),8.14(s,1H), 8.06(d,1H),7.91(d,1H),7.70(d,1H),7.51(d,1H),7.44(s,1H),6.62(q,1H),3.57(s,2H), 3.26(s,3H),2.84(d,3H),2.56(d,3H),2.41(m,8H),2.18(s,3H)。

ESI-MS m/z:[M+H]⁺=576.3。

Embodiment 93- ((fluoro- [1,2,4] triazoles of 2- amino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of fluoro- [1,2,4] triazoles of the bromo- 8- of step 12- amino -6- simultaneously [1,5-a] pyridine

With the fluoro- 5- bromopyridines of 2- amino -3- as raw material, title is prepared according to the method for 1 step 1 of embodiment and step 2 Compound.

¹H NMR(300MHz,DMSO-d₆)δ:8.30(s,1H),7.27(dd,1H),4.58(m,2H)。

Step 23- ((fluoro- [1,2,4] triazoles of 2- amino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- methyl - The preparation of N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With fluoro- [1,2,4] triazoles of the bromo- 8- of 2- amino -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, according to the method system of 1 step 7 of embodiment It is standby go out target compound.

1H NMR(300MHz,DMSO-d6)δ:10.56(s,1H),8.88(s,1H),8.19(s,1H),8.14(s,1H), 8.05(d,1H),7.91(d,1H),7.69(d,1H),7.64(d,1H),7.51(d,1H),6.44(s,2H),3.57(s,2H), 2.73(s,3H),2.40(m,8H),2.25(s,3H)。

ESI-MS m/z:[M+H]⁺=566.4。

Embodiment 103- ((2- methylamino -5- trifluoromethyls-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetylene Base) -4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- methylamino -5- trifluoromethyl -6- simultaneously [1,5-a] pyridine

With the bromo- 6- trifluoromethyl pyridines of 2- amino -5- as raw material, title is prepared according to the method for 1 step 1-3 of embodiment Product.

1H NMR(500MHz,DMSO-d6)δ：7.79-7.77（m,1H),7.63-7.61(m,1H),6.91(s,1H), 2.85(s,3H)。

ESI-MS m/z:[M+H]⁺=296.0。

Step 23- ((2- methylamino -5- trifluoromethyls-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) - The preparation of 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With 2- methylamino -5- trifluoromethyls -6- bromines [1,2,4] triazole simultaneously [1,5-a] pyridine and 3- acetenyl -4- first Base-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to 1 step 7 of embodiment Method prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.54(s,1H),8.20(d,1H),8.16(d,1H),8.05(d,1H), 7.96(dd,1H),7.77(dd,2H),7.71(d,1H),7.54(d,1H),7.02(q,1H),3.58(s,2H),2.87(d, 3H),2.55(s,3H),2.41(m,8H),2.21(s,3H)。

ESI-MS m/z:[M+H]⁺=630.3。

Embodiment 113- ((2- amino -8- methyl-carbamoyls [1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetylene Base) -4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- 8- methyl-carbamoyls [1,2,4] triazoles of step 12- amino -6- simultaneously [1,5-a] pyridine

With the bromo- N- methylnicotinamides of 2- amino -5- as raw material, prepare according to the method for 1 step 1 of embodiment and step 2 Title compound.

Step 23- ((2- amino -8- methyl-carbamoyls [1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetylene Base) -4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide preparation

With bromo- 8- methyl-carbamoyls [1,2,4] triazoles of 2- amino -6- simultaneously [1,5-a] pyridine] pyridine and 3- acetylene Base -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to embodiment The method of 1 step 7 prepares target compound.

1H NMR(300MHz,DMSO-d6)δ:10.51(s,1H),9.17(d,2H),8.20(s,2H),8.14(s,1H), 8.06(d,1H),7.93(d,1H),7.71(d,1H),7.52(d,1H),6.63(s,2H),3.57(s,2H),2.95(d,3H), 2.58(s,3H),2.40(m,8H),2.17(s,3H)。

ESI-MS m/z:[M+H]⁺=605.5。

Embodiment 123- ((chloro- [1,2,4] triazoles of 2- amino -7- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of chloro- [1,2,4] triazoles of the bromo- 7- of step 12- amino -6- simultaneously [1,5-a] pyridine

With the chloro- 5- bromines of 2- amino -4- as raw material, title compound is prepared according to the method for 1 step 1 of embodiment and step 2 Thing.

1H NMR(300MHz,DMSO-d6)δ:7.25(t,2H),7.17(d,2H)。

Step 23- ((chloro- [1,2,4] triazoles of 2- amino -7- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- methyl - The preparation of N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With chloro- [1,2,4] triazoles of the bromo- 7- of 2- amino -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, according to the method system of 1 step 7 of embodiment It is standby go out target compound.

1H NMR(300MHz,DMSO-d6)δ:10.55(s,1H),9.09(s,1H),8.20(d,1H),8.16(s,1H), 8.06(d,1H),7.94(d,1H),7.72(s,1H),7.71(s,1H),7.53(d,1H),6.36(s,2H),3.57(s,2H), 2.58(s,3H),2.35(s,3H),2.31(m,8H)。

ESI-MS m/z:[M+H]⁺=582.2。

Embodiment 133- ((2- methylamino -5- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- methylamino -5- methyl -6- simultaneously [1,5-a] pyridine

With the bromo- 6- methyl of 2- amino -5- as raw material, according to the method synthesis title product of 1 step 1-3 of embodiment.

ESI-MS m/z:[M+H]⁺=241.6。

Step 23- ((2- methylamino -5- methyl-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- methylamino -5- methyl -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.56(s,1H),8.21(s,1H),8.18(s,1H),8.07(d,1H), 7.92(d,1H),7.71(d,1H),7.61(d,1H),7.53(d,1H),7.33(d,1H),6.69(m,1H),3.58(s,2H), 2.87(m,6H),2.58(s,3H),2.42(m,8H),2.22(s,3H)。

ESI-MS m/z:[M+H]⁺=576.3。

Embodiment 143- ((chloro- [1,2,4] triazoles of 2- amino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of chloro- [1,2,4] triazoles of the bromo- 8- of step 12- amino -6- simultaneously [1,5-a] pyridine

With the chloro- 5- bromopyridines of 2- amino -3- as raw material, title is prepared according to the method for 1 step 1 of embodiment and step 2 Compound.

1H NMR(300MHz,DMSO-d6)δ:8.38(s,1H),7.59(s,1H),4.67(m,2H)。

Step 23- ((chloro- [1,2,4] triazoles of 2- amino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- methyl - The preparation of N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With chloro- [1,2,4] triazoles of the bromo- 8- of 2- amino -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, according to the method system of 1 step 7 of embodiment It is standby go out target compound.

1H NMR(300MHz,DMSO-d6)δ:10.54(s,1H),8.96(s,1H),8.21(s,1H),8.16(d,1H), 8.08(d,1H),7.93(d,1H),7.84(d,1H),7.71(d,1H),7.52(d,1H),6.45(s,2H),3.62(s,2H), 2.57(s,3H),2.45(m,8H),1.75(s,3H)。

ESI-MS m/z:[M+H]⁺=582.1。

Embodiment 153- ((fluoro- [1,2,4] triazoles of 2- dimethylamino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With the fluoro- 5- bromopyridines of 2- amino -3- as raw material, title compound is prepared according to the method for 1-2 the step of embodiment 1 Thing.

The preparation of fluoro- [1,2,4] triazoles of the bromo- 8- of step 22- dimethylamino -6- simultaneously [1,5-a] pyridine

In dry two mouthfuls of flasks of 100ml, add fluoro- [1,2, the 4] triazoles of the bromo- 8- of compound 2- amino -6- simultaneously [1, 5-a] pyridine (160mg, 0.69mmol), sodium hydride（138mg,3.47mmol）And tetrahydrofuran（10mL）.Reaction system is in nitrogen 0 DEG C under the conditions of gas shielded, react 30 minutes.Then, iodomethane（199mg,1.38mmol）It is dissolved in tetrahydrofuran（10mL）In batches It is added drop-wise in above-mentioned reactant liquor, 12h is stirred at room temperature.TLC is detected.Under condition of ice bath, add methyl alcohol that excessive sodium hydride is quenched, it is dense Contracting.Add water（20ml）, pH to 7, ethyl acetate are adjusted with 1N watery hydrochloric acid（50ml）Three times, concentration is dried, and filters, column chromatography Purifying obtains title compound.

¹H NMR(500MHz,DMSO-d₆)δ8.95（s,1H),7.78(d,1H),3.04(s,6H).

Step 33- ((fluoro- [1,2,4] triazoles of 2- dimethylamino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With fluoro- [1,2,4] triazoles of the bromo- 8- of 2- dimethylamino -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.52(s,1H),8.97(s,1H),8.20(s,1H),8.15(d,1H), 8.05(d,1H),7.93(d,1H),7.70(d,1H),7.68(d,1H),7.52(d,1H),3.57(s,2H),3.25(s,3H), 3.08(s,3H),2.56(s,3H),2.40(m,8H),2.18(s,3H)。

ESI-MS m/z:[M+H]⁺=594.3。

Embodiment 163- ((2- methylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- methyl - N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- methylamino -6- simultaneously [1,5-a] pyridine

With 2- amino -5- bromopyridines as raw material, title compound is prepared according to the method for 1 step 1-3 of embodiment.

1H NMR(300MHz,DMSO-d6)δ:8.95(d,1H),7.56(dd,1H),7.34(d,1H),6.54(d,1H), 2.81(d,3H)。

ESI-MS m/z:[M+H]⁺=227.2。

Step 23- ((2- methylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- methyl-N- The preparation of [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- methylamino -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- Methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, prepare according to the method for 1 step 7 of embodiment Target compound.

1H NMR(300MHz,DMSO-d6)δ:10.52(s,1H),8.99(s,1H),8.21(s,1H),8.16(d,1H), 8.06(d,1H),7.92(d,1H),7.71(d,1H),7.60(d,1H),7.52(d,1H),7.42(d,1H),6.64(m,1H), 3.57(s,2H),2.86(dd,3H),2.56(s,3H),2.37(m,8H),2.16(s,3H)。

ESI-MS m/z:[M+H]⁺=562.2。

Embodiment 173- ((chloro- [1,2,4] triazoles of 2- methylamino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- Methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of chloro- [1,2,4] triazoles of the bromo- 8- of step 12- methylamino -6- simultaneously [1,5-a] pyridine

With the chloro- 5- bromopyridines of 2- amino -3- as raw material, title compound is prepared according to the method for 1 step 3 of embodiment.

1H NMR(300MHz,DMSO-d6)δ:9.00(s,1H),7.88(s,1H),6.81(s,1H),2.82(d,3H)。

ESI-MS m/z:[M+H]⁺=260.9。

Step 23- ((chloro- [1,2,4] triazoles of 2- methylamino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With chloro- [1,2,4] triazoles of the bromo- 8- of 2- methylamino -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.54(s,1H),8.20(s,1H),8.15(s,1H),8.06(d,1H), 7.93(d,1H),7.73(d,1H),7.68(d,1H),7.62(d,1H),7.52(d,1H),6.94(m,1H),3.57(s,2H), 2.86(s,3H),2.73(s,3H),2.37(m,8H),2.18(s,3H)。

ESI-MS m/z:[M+H]⁺=596.2。

Embodiment 183- ((fluoro- [1,2,4] triazoles of 2- methylamino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- Methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of fluoro- [1,2,4] triazoles of the bromo- 8- of step 12- methylamino -6- simultaneously [1,5-a] pyridine

With the fluoro- 5- bromopyridines of 2- amino -3- as raw material, title compound is prepared according to the method for implementing 1 step 1-3.

1H NMR(300MHz,DMSO-d6)δ:8.34 (s, 1H), 7.37-7.35 (m, 1H), 3.49 (s, 1H), 3.06 (s, 3H)。

ESI-MS m/z:[M+H]⁺=245.0。

Step 23- ((fluoro- [1,2,4] triazoles of 2- methylamino -8- simultaneously [1,5-a] pyridine -6- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With fluoro- [1,2,4] triazoles of the bromo- 8- of 2- methylamino -6- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.54(s,1H),8.94(s,1H),8.20(s,1H),8.16(d,1H), 8.06(d,1H),7.93(d,1H),7.71(d,1H),7.66(d,1H),7.52(d,1H),6.87(m,1H),3.57(s,2H), 2.85(d,3H),2.56(s,3H),2.40(m,8H),2.19(s,3H)。

ESI-MS m/z:[M+H]⁺=580.2。

Embodiment 193- ((2- isopropylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- amino -7- simultaneously [1,5-a] pyridine

With 2- amino -4- bromopyridines as raw material, title compound is prepared according to the method for implementing 1 step 1 and step 2.

The preparation of bromo- [1,2,4] triazoles of step 22- isopropylamino -7- simultaneously [1,5-a] pyridine

In dry two mouthfuls of flasks of 50ml, bromo- [1,2, the 4] triazoles of 2- amino -7- simultaneously [1,5-a] pyridine is added (300mg, 1.4mmol), sodium hydrogen（280mg, 7.0mmol, mass fraction 60%）And DMF（5ml）.Reaction system is protected in nitrogen Under the conditions of 10 DEG C, react 1 hour.Then, isopropyl bromide（512mg,4.2mmol）It is added drop-wise in above-mentioned reactant liquor in batches, room temperature Stirring 2h.TLC is detected.Reactant liquor is poured in frozen water, ethyl acetate extraction（50ml）Three times, concentration is dried, and filters, post layer Analysis purifying obtains title compound.

¹H NMR(300MHz,DMSO-d₆)δ:8.53(d,1H),7.67(d,1H),7.01(dd,1H),6.53(d,1H), 3.82-3.73(m,1H),1.16(d,6H)。

ESI-MS m/z:[M+H]⁺=255.0。

Step 33- ((2- isopropylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- methyl - The preparation of N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- isopropylamino -7- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, according to the method system of 1 step 7 of embodiment It is standby go out target compound.

1H NMR(300MHz,DMSO-d6)δ:10.54(s,1H),8.63(d,1H),8.20(s,2H),8.06(d,1H), 7.94(d,1H),7.71(d,1H),7.62(s,1H),7.53(d,1H),7.01(d,1H),6.57(d,1H),3.85(m,1H), 3.57(s,2H),2.57(s,3H),2.39(m,8H),2.17(s,3H),1.18(s,6H)。

ESI-MS m/z:[M+H]⁺=590.2。

Embodiment 203- ((2- methylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridine -5- bases) acetenyl) -4- methyl - N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- methylamino -5- simultaneously [1,5-a] pyridine

With 2- amino -6- bromopyridines as raw material, title compound is prepared according to the method for implementing 1 step 1-3.

ESI-MS m/z:[M+H]⁺=227.0。

Step 23- ((2- methylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridine -5- bases) acetenyl) -4- methyl-N- The preparation of [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- methylamino -5- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- Methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, prepare according to the method for 1 step 7 of embodiment Target compound.

1H NMR(300MHz,DMSO-d6)δ:10.58(s,1H),8.25(s,1H),8.21(d,1H),8.06(d,1H), 8.02(d,1H),7.72(d,1H),7.58(d,1H),7.48(d,2H),7.27(d,1H),6.61(m,1H),3.58(s,2H), 2.88(dd,3H),2.68(s,3H),2.40(m,8H),2.19(s,3H)。

ESI-MS m/z:[M+H]⁺=562.3。

Embodiment 213- ((2- methylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridine -8- bases) acetenyl) -4- methyl - N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 12- methylamino -8- simultaneously [1,5-a] pyridine

With 2- amino -3- bromopyridines as raw material, title compound is prepared according to the method for implementing 1 step 1 to step 3.

¹H NMR(300MHz,DMSO-d₆)δ:8.62(d,1H),7.73(d,1H),6.82-6.77(m,1H),6.69- 6.67(m,1H),2.82(d,3H)。

ESI-MS m/z:[M+H]⁺=227.0。

Step 23- ((2- methylaminos-[1,2,4] triazole simultaneously [1,5-a] pyridine -8- bases) acetenyl) -4- methyl-N- The preparation of [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- methylamino -8- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- Methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, prepare according to the method for 1 step 7 of embodiment Target compound.

1H NMR(300MHz,DMSO-d6)δ:10.56(s,1H),8.68(d,1H),8.21(s,1H),8.18(d,1H), 8.06(d,1H),7.94(d,1H),7.73(s,1H),7.70(s,1H),7.54(d,1H),6.93(d,1H),6.68(m,1H), 3.57(s,2H),2.85(d,3H),2.61(s,3H),2.40(m,8H),2.19(s,3H)。

ESI-MS m/z:[M+H]⁺=562.3。

Embodiment 223- ((2- (2,2,2- trifluoroacetamidos) imidazo [1,2-a] pyridin-7-yl) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of step 12- tolysulfonyl amino -4- bromopyridines

2- amino -4- bromopyridines (1.72g, 10mmol), pyridine (5mL), ice bath are added in a 50mL round-bottomed flask Under be slowly added dropwise paratoluensulfonyl chloride（2.29g,12mmol）, 90 DEG C are warming up to, 6h is reacted, water, ethyl acetate extraction, silicon is added Plastic column chromatography obtains title compound.

The preparation of step 22- (bromo- -1 (the 2H)-yls of 2 tolysulfonyl imines yl pyridines of 4-) acetamide

2- tolysulfonyl amino -4- bromopyridines (327mg, 1.0mmol), DMF are added in 25mL round-bottomed flasks (5Ml), 2- acetbromamides（152mg,1.1mmol）And diisopropyl ethyl amine（142mg,1.1mmol）, room temperature reaction 24h. Water, ethyl acetate is added to extract, anhydrous sodium sulfate drying, silica gel column chromatography obtain title compound.

Step 32- (2,2,2- trifluoroacetamidos) -7- bromo- imidazo [1,2-a] pyridines

2- is added in 25mL round-bottomed flasks（Bromo- -1 (the 2H)-bases of 2 tolysulfonyl imines yl pyridines of 4-）Acetamide (385mg, 1.0mmol), TFAA (2.10g, 10mmol), and dichloromethane（5mL）, flow back 2h, TLC monitoring reactions Finish, under ice bath, add water, dichloromethane to extract 3 times, merge organic phase, saturated sodium bicarbonate solution and saturated common salt washing Wash, anhydrous sodium sulfate drying, silica gel column chromatography obtain title compound.

1H NMR(300MHz,DMSO-d6)δ:12.51(s,1H),8.57(d,1H),8.28(s,1H),7.85(s,1H), 7.14(dd,1H)。

ESI-MS m/z:[M+H]⁺=329.9。

Step 43- ((2- (2,2,2- trifluoroacetamide amino) imidazo [1,2-a] pyrazine -7- bases) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With the 2- bromo- imidazos of (2,2,2- trifluoroacetamidos) -7- [1,2-a] pyridines and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide is raw material, according to the method for 1 step 7 of embodiment Prepare target compound.

1H NMR(300MHz,DMSO-d6)δ:10.50(s,1H),8.63(d,1H),8.31(s,1H),8.17(s,2H), 8.03(d,1H),7.91(d,1H),7.78(d,1H),7.69(s,1H),7.67(s,1H),7.50(d,1H),7.07(d,1H), 3.54(s,2H),2.55(s,3H),2.36(m,8H),2.16(s,3H)。

ESI-MS m/z:[M+H]⁺=643.3。

Embodiment 233- ((2- acetylaminohydroxyphenylarsonic acids [1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With 2- amino -4- bromopyridines as raw material, title compound is prepared according to the method for 1 step 1 of embodiment and step 2 Thing.

The preparation of bromo- [1,2,4] triazoles of step 22- acetylaminohydroxyphenylarsonic acid 7- simultaneously [1,5-a] pyridine

In dry two mouthfuls of flasks of 50ml, bromo- [1,2, the 4] triazoles of compound 2- amino -7- simultaneously [1,5-a] pyrrole is added Pyridine (213mg, 1mmol) and acetic anhydride（10ml）, it is subsequently added into pyridine (118mg, 1.5mmol).85 DEG C of backflows of reaction system, instead Should overnight.TLC detects that reactant liquor concentration, ethyl acetate/water extraction, organic layer are dried, and column chromatography purifying obtains title compound Thing.

ESI-MS m/z:[M+H]⁺=255.0。

Step 33- ((2- acetylaminohydroxyphenylarsonic acids [1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- methyl-N- The preparation of [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- acetylaminohydroxyphenylarsonic acid 7- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, according to the method system of 1 step 7 of embodiment It is standby go out target compound.

1H NMR(300MHz,DMSO-d6)δ:10.84(s,1H),10.54(s,1H),8.90(d,1H),8.22(d, 1H),8.21(d,1H),8.07(d,1H),7.98(d,1H),7.97(s,1H),7.71(d,1H),7.55(s,1H),7.27 (dd,1H),3.58(s,2H),2.60(s,3H),2.41(m,8H),2.19(s,3H),2.16(s,3H)。

ESI-MS m/z:[M+H]⁺=590.3。

Embodiment 243- ((2- methanesulfonamidos-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- first Base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazoles of step 22- methanesulfonamido -7- simultaneously [1,5-a] pyridine

In dry two mouthfuls of flasks of 100mL, be initially charged THF (5mL), be subsequently adding compound 2- amino -7- it is bromo- [1, 2,4] triazole simultaneously [1,5-a] pyridine (213mg, 100mmol), mesyl chloride（3ml）, pyridine（5mL）.Room temperature reaction 24 is little When.TLC detection raw materials disappear.Add water（10ml）, dichloromethane（20ml）Extraction three times, concentration are dried, and filter, column chromatography Purify to obtain title compound.

1H NMR(300MHz,DMSO-d6)δ:9.01(d,1H),8.35(s,1H),7.58(dd,1H),3.68(s,3H)。

Step 33- ((2- methanesulfonamidos-[1,2,4] triazole simultaneously [1,5-a] pyridin-7-yl) acetenyl) -4- methyl - The preparation of N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 2- methanesulfonamido -7- simultaneously [1,5-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, according to the method system of 1 step 7 of embodiment It is standby go out target compound.

1H NMR(300MHz,DMSO-d6)δ:10.53(s,1H),8.81(d,1H),8.21(d,2H),8.06(d,1H), 7.95(d,1H),7.88(s,1H),7.71(d,1H),7.54(d,1H),7.21(d,1H),3.58(s,2H),3.26(s,3H), 2.96(b,1H),2.59(s,3H),2.42(m,8H),2.22(s,3H)。

ESI-MS m/z:[M+H]⁺=626.2。

Embodiment 253- ((3- oxo -2,3- dihydros-[1,2,4] triazole simultaneously [4,3-a] pyridine -8- bases) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of step 12- diazanyl -3- bromopyridines

Add the fluoro- 3- bromopyridines (3.52g, 20mmol) of 2- in 250ml round-bottomed flask, hydrazine hydrate (200mmol) and THF (25mL), it is 30% 6h. to be stirred at room temperature and is concentrated into volume, and cooling crystallization under ice bath is filtered, and collects crystal, and vacuum drying is obtained To title compound.

The preparation of bromo- [1,2,4] triazole of step 28- simultaneously [4,3-a] pyridine -3 (2H) -one

In dry two mouthfuls of flasks of 100ml, triphosgene (888mg, 300mmol), tetrahydrofuran solution 100mL. are added It is dividedly in some parts compound 2- diazanyl -3- bromopyridines（188mg, 100mmol）, reacting 3 hours, TLC points board raw material disappears.In ice bath Under be slowly added to 100mL water.Vacuum distillation removes 70% solvent.Then stand slowly precipitation solid filtration to drain,

Forced air drying obtains title compound in 8 hours.

Step 33- ((3- oxo -2,3- dihydros-[1,2,4] triazole simultaneously [4,3-a] pyridine -8- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 8- simultaneously -3 (2H) -one of [4,3-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- Methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, prepare according to the method for 1 step 7 of embodiment Target compound.

1H NMR(300MHz,DMSO-d6)δ:12.67(s,1H),10.58(s,1H),8.21(s,1H),8.18(s, 1H),8.07(d,1H),7.95(d,1H),7.91(d,1H),7.71(d,1H),7.55(d,1H),7.54(s,1H),6.65(t, 1H),3.58(s,2H),2.59(s,3H),2.42(m,8H),2.22(s,3H)。

ESI-MS m/z:[M+H]⁺=549.3。

Embodiment 263- ((3- oxo -2,3- dihydros-[1,2,4] triazole simultaneously [4,3-a] pyridin-7-yl) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazole of step 17- simultaneously [4,3-a] pyridine -3 (2H) -one

With the fluoro- 4- bromopyridines of 2- as raw material, title compound is prepared with the method for 25 step 1 of embodiment and step 2.

ESI-MS m/z:[M+H]⁺=214.0。

Step 23- ((3- oxo -2,3- dihydros-[1,2,4] triazole simultaneously [4,3-a] pyridin-7-yl) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 7- simultaneously -3 (2H) -one of [4,3-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- Methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, prepare according to the method for 1 step 7 of embodiment Target compound.

1H NMR(300MHz,DMSO-d6)δ:12.66(s,1H),10.52(s,1H),8.19(d,2H),8.06(d, 1H),7.95(d,1H),7.85(d,1H),7.71(d,1H),7.54(t,2H),6.62(d,1H),3.58(s,2H),2.56(s, 3H),2.42(m,8H),2.36(s,3H)。

ESI-MS m/z:[M+H]⁺=549.2。

Embodiment 273- ((3- oxo -2,3- dihydros-[1,2,4] triazole simultaneously [4,3-a] pyridine -5- bases) acetenyl) - 4- methyl-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

The preparation of bromo- [1,2,4] triazole of step 15- simultaneously [4,3-a] pyridine -3 (2H) -one

With the fluoro- 6- bromopyridines of 2- as raw material, title compound is prepared with the method for 25 step 1 of embodiment and step 2.

ESI-MS m/z:[M+H]⁺=214.0。

Step 23- ((3- oxo -2,3- dihydros-[1,2,4] triazole simultaneously [4,3-a] pyridine -5- bases) acetenyl) -4- first The preparation of base-N- [4- ((4- methylpiperazine-1-yls) methyl) -3- trifluoromethyls] benzamide

With bromo- [1,2,4] triazoles of 5- simultaneously -3 (2H) -one of [4,3-a] pyridine and 3- acetenyl -4- methyl-N- [4- (4- Methylpiperazine-1-yl methyl) -3- trifluoromethyls] benzamide be raw material, prepare according to the method for 1 step 7 of embodiment Target compound.

1H NMR(300MHz,DMSO-d6)δ:12.68(s,1H),10.59(s,1H),8.22(s,1H),8.18(s, 1H),8.08(d,1H),7.96(d,1H),7.91(d,1H),7.71(d,1H),7.55(d,1H),7.54(d,1H),6.65(t, 1H),3.61(s,2H),2.59(s,3H),2.42(m,8H),2.22(s,3H)。

ESI-MS m/z:[M+H]⁺=549.3。

Embodiment 283- ((2- methylamino imidazo [1,2-a] pyridin-7-yls) acetenyl) -4- methyl-N- [4- ((4- first Base piperazine -1- bases) methyl) -3- trifluoromethyls] benzamide

The preparation of step 12- methylamino -7- bromo- imidazo [1,2-a] pyridines

2- (2,2,2- trifluoroacetamidos) -7- bromo- imidazo [1,2-a] pyridines are added in 100mL round-bottomed flasks (308mg, 1.0mmol), iodomethane (170mg, 1.2mmol), potassium carbonate (414mg, 3.0mmol) and acetone (5mL), room temperature Stirring 14h.Water is added in system（5mL), flow back 2h, and removal of solvent under reduced pressure adds water, is extracted with ethyl acetate, anhydrous sulphur Sour sodium is dried, and silica gel column chromatography obtains target product.

1H NMR(300MHz,DMSO-d6)δ:8.71(d,1H),8.34(s,1H),8.30(s,1H),7.54(dd,1H), 3.76(s,3H),2.67(b,1H)。

ESI-MS m/z:[M+H]⁺=226.0。

Step 23- ((2- methylamino imidazo [1,2-a] pyridin-7-yls) acetenyl) -4- methyl-N- [4- ((4- methyl Piperazine -1- bases) methyl) -3- trifluoromethyls] and benzamide preparation

With bromo- imidazo [1,2-a] pyridines of 2- methylamino -7- and 3- acetenyl -4- methyl-N- [4- (4- methyl piperazines - 1- ylmethyls) -3- trifluoromethyls] benzamide be raw material, prepare target chemical combination according to the method for 1 step 7 of embodiment Thing.

1H NMR(300MHz,DMSO-d6)δ:10.56(s,1H),8.82(d,1H),8.39(s,1H),8.23(d,2H), 8.22(s,1H),8.07(d,1H),7.98(d,1H),7.72(d,1H),7.57(d,1H),7.49(d,1H),4.03(q,1H), 3.83(s,3H),3.58(s,2H),2.62(s,3H),2.40(m,8H),2.17(s,3H)。

ESI-MS m/z:[M+H]⁺=561.4。

The Compound ira vitro cytoactive of 1 present invention of experimental example is evaluated

1. test material

1.1 compound

The compound of the present invention prepared by above example, each compound DMSO is dissolved to after 10mM, with training completely Foster base is diluted to 50 μM, then with containing 0.1%DMSO complete medium be diluted to 10 μM after, successively 10 times dilution, totally 10 it is dense Degree.

1.2 cell

K562 leukaemia：Purchased from ATCC companies of the U.S.

1.3 reagent

Dimethyl sulfoxide (DMSO)（Dimethyl sulfoxide, DMSO）, it is purchased from Sigma Co., USA；

Luminescence method cell viability detection kit Luminescent Cell Viability Assay Kit）, it is purchased from Promega companies of the U.S.；

IMEM culture mediums（IMEM medium）, it is purchased from Gibco companies of the U.S.；

Penicillin/streptomycin（Pen/Strep）, it is purchased from Gibco companies of the U.S.；

Hyclone（Fatal bovine serun, FBS）, it is purchased from Gibco companies of the U.S.；

0.25% pancreatin containing EDTA（0.25%Trypsin-EDTA）, it is purchased from Gibco companies of the U.S.；

10cm Tissue Culture Dish（10cm cell culture dish）, it is purchased from Corning companies of the U.S.；

50mL centrifuge tubes（50mL centrifuge tube）, it is purchased from Corning companies of the U.S.；

The flat printing opacity blank in 384 holes（384Well Flat Clear Bottom White）, it is purchased from U.S. Corning public Department；

Phosphate buffer（Phosphate Buffered Saline, PBS）, prepare weekly.

1.4 instrument

Automatic confocal fluorescence multi-function microplate reader（PHERAstar Plus）, it is purchased from German BMG Labtech companies.

2. experimental technique：

1) logarithmic phase cell, adjustment concentration of cell suspension to 1 × 10 are collected⁵Individual/ml, 384 orifice plates add 40 μ L thin per hole Born of the same parents' suspension, is 4 × 10 per hole cell number³Individual/hole.Edge hole is filled with aseptic PBS；

2) add the compound of the present invention of the concentrations above gradient of 10 μ L.Each compound each concentration is in triplicate. Blank adds the DMSO of 10 μ L comparable sodiums；

3) cell is in 37 DEG C/5%CO₂It is incubated in incubator；

4) 30 μ L luminescence method cell viability detection reagent mixed liquors are added after dosing 72h；

5)37℃/5%CO₂10min is incubated in incubator；Determine chemical on PHERAstar ELIASAs after slow-speed of revolution centrifugation Luminous value；

6) cell viability（Cell Viability）=（RLU_Sample/RLU_{It is negative}）× 100%, wherein RLU_SampleFor medicine feeding hole RLU （Relative light unit）Value, RLU_{It is negative}For not medicine feeding hole RLU values（That is the cell controls of comparable sodium DMSO process）, adopt Four parameter logistic fit module of Graphpad Prism4.0 data processing softwares carries out processing data and calculates IC₅₀。IC₅₀Value is represented With not plus compared with compound treatment group, compound suppresses the corresponding compound concentration of 50% cell growth.IC₅₀The results are shown in Table 1.

Table 1

"-" is represented does not test.

IC of the compound of the present invention to leukaemia be can be seen that from above experimental result₅₀Value has in nM levels Strong inhibitory activity.Jia H-Y etc.（Jia H-Y,et al.ZD6474inhibits Src kinase leading to apoptosis of imatinib-resistant K562cells.Leuk Res(2009),doi:10.1016/ j.leukres.2009.03.033）Research shows IC of the Imatinib to K562 cells₅₀It is worth for 280nM, H Luo etc.（H Luo, et al.HH-GV-678,a novel selective inhibitor of Bcr-Abl,outperforms imatinib and effectively overrides imatinib resistance.Leukemia (2010)24,1807–1809; doi:10.1038/leu.2010.169;Published online12August2010）Experiment proves Imatinib to K562 The IC of cell₅₀It is worth for 296nM.As can be seen here, for the leukaemia of not resistance, compound and the Imatinib of the present invention have Have quite, even more excellent effect.

Experimental example 2ABL1 (T315I) tyrosine kinase activity evaluation

The suppression of compound prepared by this experiment test embodiment of the present invention to ABL (T315I) kinase activity, using her horse For Buddhist nun as control.Imatinib is obtained with reference to the method described in Chinese patent CN1043531C and is reflected by hydrogen spectrum and mass spectrum It is fixed.

Using commercially available people source ABL T315I mutant enzymes（Human ABL1 (T315I), active, catalog number (Cat.No.) #14-522, Millipore companies, the U.S.）Test ABL (T315I) tyrosine kinase activity.Kinase activity measure is carried out by manufacturers instruction. Peptide substrates（Peptide substrate）For Abltide (EAIYAAPFAKKK), Millipore companies of the U.S. are purchased from.Ion is handed over Change chromatography filter paper P81（ion exchange filter paper）It is purchased from Whatman companies of Britain.[γ -33P] ATP is purchased from Perkin Elmer companies.

The compound and Imatinib of the present invention starts respectively 3 times of serial dilutions from 1 μM, totally 10 concentration (50.8pM, 152.0pM, 457.0pM, 1.37nM, 4.12nM, 12.3nM, 37.0nM, 111.0nM, 333.0nM and 1.0 μM).Add per hole 5.0 μM of Abltide, are subsequently adding people source T315I mutant enzymes.Add under room temperature [γ-³³P] ATP, final concentration of 1.0 μM, reaction 120 minutes.20 μ l aliquots are transferred on P81 ion-exchange chromatography paper.Then chromatographic paper is abundant with 0.75% phosphoric acid solution Washing 3 times, then washed once with acetone.Finally, carry out γ-³³P radioactivity determinations.Experimental result is shown in Table 2.

Table 2

Above test result indicate that, the IC of the compound to T315I mutant enzymes of the present invention₅₀Imatinib is significantly better than, is had There is the ability of potent suppression T315I mutant enzymes.

Can draw from the experimental result of the present invention, the compound of the present invention is not only to having the leukaemia being mutated tool There is extraordinary effect, and T315I mutant enzymes can be significantly inhibited, therefore be the BCR-ABL inhibitor of wide spectrum.For to junket Histidine kinase inhibitor（TKI）Treatment resistance or the tumor patients in heilongjiang of opposing, such as chronic myelocytic leukemia (CML) are chronic Phase, CML-BC, accelerated period patient and Philadelphia Chromosome Positive（Ph+）Chronic myelocytic leukemia and acute lymphoblastic it is white The prospect that blood patient should have.

Although being below described in detail to the present invention, however it is understood by skilled practitioners that without departing from this Various modifications and changes can be carried out to the present invention on the premise of bright spirit and scope.The interest field of the present invention is not limited to The detailed description made above, and claims should be belonged to.

Claims

1. a kind of compound or its pharmaceutically acceptable salt, wherein the compound is selected from following compound：

2. a kind of pharmaceutical composition, which includes the compound described in claim 1 or its pharmaceutically acceptable salt and pharmaceutically acceptable load Body.

3. the pharmaceutical composition described in compound described in claim 1 or its pharmaceutically acceptable salt or claim 2 is in system The application being ready for use in the medicine for the treatment of or prevention of tumor.