CN104583195B - Nitrogenous heteroaromatic ring derivative as tyrosine kinase inhibitor - Google Patents

Nitrogenous heteroaromatic ring derivative as tyrosine kinase inhibitor Download PDF

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CN104583195B
CN104583195B CN201380041846.3A CN201380041846A CN104583195B CN 104583195 B CN104583195 B CN 104583195B CN 201380041846 A CN201380041846 A CN 201380041846A CN 104583195 B CN104583195 B CN 104583195B
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CN104583195A (en
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罗浩贤
张艳
张倩
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SHANDONG HENRY MEDICAL SCIENCE AND TECHNOLOGY Co Ltd
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    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/4025—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil not condensed and containing further heterocyclic rings, e.g. cromakalim
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    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
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    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
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    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
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    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
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    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links

Abstract

The present invention relates to general formula (I) compounds represented as tyrosine kinase inhibitor, the preparation method of these compounds, pharmaceutical composition containing these compounds, application of these compounds in preventing and/or treating individual in the relevant leukemia of B cell, inflammatory and autoimmune disease.

Description

Nitrogenous heteroaromatic ring derivative as tyrosine kinase inhibitor
1, technical field
The invention belongs to pharmaceutical technology fields, and in particular to the nitrogenous hetero-aromatic ring as tyrosine kinase inhibitor derives Object, its pharmaceutically acceptable salt or its stereoisomer, the preparation method of these compounds, the drug containing these compounds Composition prevents using these compounds and/or treats (such as the B cell chronic lymphocytic of the relevant leukemia of B cell in individual Cancer, non-Hodgkin lymphoma), inflammatory and autoimmune disease (such as rheumatoid arthritis, systemic loupus erythematosus Deng) method and these compounds for preventing and/or treating the relevant leukemia of B cell, (such as B cell is chronic preparing Lymphocyte cancer, non-Hodgkin lymphoma), inflammatory and autoimmune disease (such as rheumatoid arthritis, systemic red Yabbi sore etc.) drug in application.
2, background technology
Protein kinase forms one of the maximum family of mankind's enzyme, and is adjusted on phosphate group to protein by adding Save many different signal transduction process (T.Hunter, Cell 1,987 50：823-829).Particularly, tyrosine kinase phosphoric acid Hydroxylic moiety of the change protein in tyrosine residue.Family tyrosine kinase include control cell growth, migration and differentiation at Member.Abnormal kinase activity has been directed to many human diseases, including cancer, autoimmune disease and inflammatory disease.Due to egg White matter kinases belongs to the key regulator of cellular signal transduction, their offers adjust cell function with small molecule kinase inhibitors Target, and therefore, it has become good drug design targets.In addition to the treatment of kinase mediated lysis, kinase activity Selectivity and effective inhibitor can also be used to study cell signaling processes and identify other cells with therapeutic potential Target.
About key effect of the B cell in the pathogenesis of autoimmunity and/or inflammatory disease, there are good evidences. Consume being closed for such as rheumatoid of inflammatory disease caused by autoantibody based on the therapeutic agent of protein such as Rituxan for B cell Section inflammation is effective (Rastetter etc., Annu Rev Med 2,004 55：477).Therefore, it plays a role in B cell activation The inhibitor of protein kinase should be that the disease pathology such as autoantibody mediated for B cell generates useful therapeutic agent.
A series of B cell responses are controlled by the signal transduction of B-cell receptor (BCR), including are proliferated and break up maturation Antibody-producting cell.BCR is the active crucial point of adjustment of B cell and abnormal signal transduction can cause the B of imbalance thin Born of the same parents are proliferated and the formation of pathogenic autoantibodies, lead to various autoimmune disease and/or inflammatory disease.Bu Ludun (Bruton's) tyrosine protein kinase (Btk) is in the film proximal end of BCR and the immediately relevant kinases of non-BCR in downstream.Btk's Shortage, which has shown that, blocks BCR signal transductions, and therefore the inhibition of Btk can be the lysis for blocking B cell to mediate Effective therapy.
Btk is the member of tyrosine kinase Tec families, and shows it is that early stage B cell is formed and mature B cell activates With key regulator (Khan etc., Immunity 1,995 3 of survival：283；Ellmeier etc., J.Exp.Med.2000 192： 1611).The Btk mutation of people cause the chain gamma-globulins of illness X to lack mass formed by blood stasis (XLA) (Lindvall etc. Immunol.Rev.2005203：200).These patients are immunocompromised hosts, and show that impaired B cell is ripe, reduction Immunoglobulin and periphery b cell level, the immune response for not depending on T cell of reduction and in the post-stimulatory decreases of BCR Calciokinesis.
Evidence about effects of the Btk in autoimmune disease and inflammatory disease is via Btk- deficient mice models It provides.In the preclinical mouse model of systemic loupus erythematosus (SLE), Btk deficient mices show significantly changing for progression of disease It is kind.In addition, Btk- deficient mices (Jansson and Holmdahl resistant to Collagen-Induced Arthritis Clin.Exp.Immunol.1993 94：459).Dosage of the verified selectivity Btk inhibitor in arthritis mouse model Dependence effect (Z.Pan etc., Chem.Med Chem.2007 2：58).
Btk also has the cell expression that may relate to lysis other than B cell.Such as Btk is by mast cell-expressed And the mast cell of Btk deficiency derived from bone marrow shows threshing (Iwaki etc. of impaired antigen induction J.Biol.Chem.2005 280：40261).This display Btk can be used for treating pathologic mast cells reaction such as allergy And asthma.In addition, the monocyte from XLA patient for wherein lacking Btk activity shows that the TNF α of reduction after stimulation generates (the J Exp such as Horwood Med 2,003 197：1603).Therefore, the inflammation that TNF α mediates can be by small molecule Btk inhibitor tune Section.Moreover it has been reported that Btk play a role in Apoptosis (IsIam and Smith Immunol.Rev.2000 178： 49), and therefore Btk inhibitor will be effective (Feldhahn etc. for treating certain B cell lymphomas and leukaemia J.Exp.Med.2005 201：1837).
The Dasatinib of listing in 2006 is multiple target point inhibitor, has Btk compared with high inhibition effect, slow for treating Property myelomatosis；In addition it is also multiple target point inhibitor to be in the PCI-32765 that the clinic III phases are studied, to Btk inhibiting effect For irreversibility, for treating lymthoma, leukaemia and autoimmune disease.
Not yet selective Btk inhibitor listing at present, the most fast drug of research are CC-292 (also known as AVL-292), In clinical I phase research, irreversible selective depression Btk, for treating leukaemia and autoimmune disease.
It is thin it can be used to prevent and/or treat B the object of the present invention is to provide excellent highly selective Btk inhibitor The relevant leukemia of born of the same parents, inflammatory and/or autoimmune disease.
3, invention content
The present invention provides the nitrogenous heteroaromatic ring derivative as tyrosine kinase inhibitor, the compound is excellent Btk inhibitor, and can be used in preventing and/or treating the relevant leukemia of B cell, inflammatory and/or autoimmune disease.
Specifically, the present invention provides the following general formula (I) compound represented, its pharmaceutically acceptable salt or it is vertical Body isomers：
Wherein, the expression phenyl of ring A and ring B independently, 3-7 member naphthenic base are miscellaneous containing the heteroatomic 3-7 members of N, O, S Naphthenic base, 4-7 unit's heteroaryls or 6-10 member bicyclic ring structures；
L1And L2Expression covalent bond independently ,-NH- ,-N (C1-3Alkyl)-,-O- ,-S (O)m,-N (C1-3Alkyl) C (O)-,-C (O) N (C1-3Alkyl)-,-N (C1-3Alkyl) S (O)2Or-S (O)2N(C1-3Alkyl)-；
A indicates covalent bond, unsubstituted or by C1-4Alkyl-substituted imino group；
B expression-CO- or-SO2-；
C indicates 1,3- allylidenes that are unsubstituted or being replaced by one or two methyl or trifluoromethyl, 1,1- or 1,2- Ethenylidene, ethynylene or 1,3-butadiene-Isosorbide-5-Nitrae-that is unsubstituted or being replaced by one to four methyl or trifluoromethyl Subunit；
D indicates covalent bond or C1-6Alkylidene；
E indicates hydrogen atom, C1-4Alkoxy, amino, 3-7 member naphthenic base, 6-10 member bicyclic ring structures, C1-4Alkyl amino or Two-(C1-4Alkyl) amino, wherein moieties may be the same or different；
R1And R3Expression hydrogen atom independently, halogen atom, cyano, nitro, C2-4Alkenyl, C2-4Alkynyl or-L3-R4,
L3Indicate covalent bond ,-NH- ,-N (C1-3Alkyl)-,-O- ,-O-C1-3Alkylidene-,-S-C1-3Alkylidene ,-S (O)m,-C (O)-,-NHC (O)-,-N (C1-3Alkyl) C (O)-,-C (O) NH- ,-C (O) N (C1-3Alkyl)-,-NHS (O)2,-N (C1-3Alkyl) S (O)2,-S (O)2NH- ,-S (O)2N(C1-3Alkyl)-,-OC (O)-or-C (O) O-,
R4Indicate hydrogen atom, C1-4Alkyl ,-N (C1-3Alkyl)2,-NHC (O) O- (C1-4Alkyl) ,-OH ,-O (C1-4Alkyl) ,- S(O)2(C1-3Alkyl), 3-7 member naphthenic base, phenyl or 5-6 unit's heteroaryls；
R2Expression-L4-R5,
L4Indicate covalent bond ,-NH- ,-N (C1-3Alkyl)-,-O-C1-3Alkylidene-,-S-C1-3Alkylidene-or-S (O)m,
R5Indicate hydrogen atom, C2-4Alkenyl, C2-4Alkynyl, C1-4Alkyl, amino ,-NH (C1-3Alkyl) ,-N (C1-3Alkyl)2,- OH ,-O (C1-3Alkyl) ,-C (O) (C1-3Alkyl) or 3-7 member naphthenic base,
Wherein L4For covalent bond when, R5Cannot be C1-4Alkyl ,-OH ,-O (C1-3Alkyl) ,-C (O) (C1-3Alkyl),
L4For-O-C1-3Alkylidene-when, R5It cannot be hydrogen atom；
The C1-4Moieties, naphthenic base, heteroaryl can be further by 1 to 4 Q1Substitution,
Q1Indicate halogen atom, C1-3Alkyl, amino, C1-3Alkyl amino, two-(C1-3Alkyl) amino, hydroxyl, C1-3Alcoxyl Base, C1-3Alkoxy carbonyl group, carbamoyl, C1-3Alkyl-carbamoyl, two-(C1-3Alkyl) carbamoyl or 3-6 member cycloalkanes Base, wherein Q1It can be identical or different；
Carbon atom can be by 1-4 identical or different N, NH, N (C in the naphthenic base, bicyclic ring structures1-3Alkyl), O, S (O)m, C (O) replace；
The heteroaryl contains 1-4 hetero atom, is independently selected from N, O or S；
M indicates 0,1 or 2；
The expressions 0,1,2,3 or 4 of p and q independently.
In a preferred embodiment, the present invention provides above-mentioned logical formula (I) compound represented, it pharmaceutically may be used The salt of receiving or its stereoisomer, wherein：
The expression phenyl of ring A and ring B independently, 3-7 member naphthenic base contain the heteroatomic 3-7 circle heterocyclic rings alkane of N, O, S Base, 4-7 unit's heteroaryls or 6-10 member bicyclic ring structures；
L1And L2Expression covalent bond independently ,-NH- ,-N (C1-3Alkyl)-,-O- ,-S (O)m,-N (C1-3Alkyl) C (O)-,-C (O) N (C1-3Alkyl)-,-N (C1-3Alkyl) S (O)2Or-S (O)2N(C1-3Alkyl)-；
A indicates covalent bond, unsubstituted or by C1-4Alkyl-substituted imino group；
B expression-CO- or-SO2-；
1,3- allylidenes, 1, the 1- or 1 that c indicates unsubstituted or replaces by one or two methyl or through trifluoromethyl, 2- ethenylidenes, ethynylene or 1,3-butadiene-that is unsubstituted or replacing by one to four methyl or through trifluoromethyl Isosorbide-5-Nitrae-subunit；
D indicates covalent bond or C1-6Alkylidene；
E indicates hydrogen atom, C1-4Alkoxy, amino, 3-7 member naphthenic base, 6-10 member bicyclic ring structures, C1-4Alkyl amino or Two-(C1-4Alkyl) amino, wherein moieties may be the same or different；
R1And R3Expression hydrogen atom independently, halogen atom, cyano, nitro, C2-4Alkenyl, C2-4Alkynyl or-L3-R4,
L3Indicate covalent bond ,-NH- ,-N (C1-3Alkyl)-,-O- ,-O-C1-3Alkylidene-,-S-C1-3Alkylidene ,-S (O)m,-C (O)-,-NHC (O)-,-N (C1-3Alkyl) C (O)-,-C (O) NH- ,-C (O) N (C1-3Alkyl)-,-NHS (O)2,-N (C1-3Alkyl) S (O)2,-S (O)2NH- ,-S (O)2N(C1-3Alkyl)-,-OC (O)-or-C (O) O-,
R4Indicate hydrogen atom, C1-4Alkyl ,-N (C1-3Alkyl)2,-NHC (O) O- (C1-4Alkyl) ,-OH ,-O (C1-4Alkyl) ,- S(O)2(C1-3Alkyl), 3-7 member naphthenic base, phenyl or 5-6 unit's heteroaryls；
R2Expression-L4-R5,
L4Expression-S-C1-3Alkylidene-or-S (O)m,
R5Indicate hydrogen atom, C2-4Alkenyl, C2-4Alkynyl, C1-4Alkyl, amino ,-NH (C1-3Alkyl) ,-N (C1-3Alkyl)2,- OH ,-O (C1-3Alkyl) ,-C (O) (C1-3Alkyl) or 3-7 member naphthenic base,
M indicates 0,1 or 2.
The C1-4Moieties, naphthenic base, heteroaryl can be further by 1 to 4 Q1Substitution,
Q1Indicate halogen atom, C1-3Alkyl, amino, C1-3Alkyl amino, two-(C1-3Alkyl) amino, hydroxyl, C1-3Alcoxyl Base, C1-3Alkoxy carbonyl group, carbamoyl, C1-3Alkyl-carbamoyl, two-(C1-3Alkyl) carbamoyl or 3-6 member cycloalkanes Base, wherein Q1It can be identical or different；
Carbon atom can be by 1-4 identical or different N, NH, N (C in the naphthenic base, bicyclic ring structures1-3Alkyl), O, S (O)m, C (O) replace；
The heteroaryl contains 1-4 hetero atom, is independently selected from N, O or S；
M indicates 0,1 or 2；
The expressions 0,1,2,3 or 4 of p and q independently.
In another preferred embodiment, the present invention provides above-mentioned logical formula (I) compound represented, its pharmaceutically Acceptable salt or its stereoisomer, wherein：
The expression phenyl of ring A and ring B independently, 5-6 member naphthenic base contain the heteroatomic 5-6 circle heterocyclic rings alkane of N, O, S Base, 5-6 unit's heteroaryls or 8-10 member bicyclic ring structures；
L1And L2Expression covalent bond independently ,-NH- ,-N (CH3)-,-O- ,-S (O)m,-N (CH3) C (O)-,-C (O) N(CH3)-,-N (CH3)S(O)2Or-S (O)2N(CH3)-；
A indicates covalent bond, unsubstituted or by CH3Substituted imino group；
B expression-CO- or-SO2-；
C indicates unsubstituted or by one or two methyl substituted 1,2- ethenylidenes or ethynylene；
D indicates covalent bond or methylene；
E indicates hydrogen atom, methoxyl group, amino, piperidyl, morpholinyl, pyrrolidinyl, piperazinyl, N methyl piperazine base, 6- 9 yuan of spirane structures, 6-8 members and ring structure, 6-8 member caged scaffolds, methylamino or two-(methyl) amino；
R1And R3Expression hydrogen atom independently, halogen atom, nitro or-L3-R4,
L3Indicate covalent bond ,-NH- ,-N (C1-3Alkyl)-,-O- ,-O-C1-3Alkylidene-,-S-C1-3Alkylidene ,-S (O)m,-C (O)-,-NHC (O)-,-C (O) NH- ,-NHS (O)2,-S (O)2NH- ,-OC (O)-or-C (O) O-,
R4Indicate hydrogen atom, C1-4Alkyl ,-N (C1-3Alkyl)2,-NHC (O) O- (C1-4Alkyl) ,-OH ,-O (C1-4Alkyl) ,- S(O)2(C1-3Alkyl), 5-6 member naphthenic base, phenyl or 5-6 unit's heteroaryls；
R2Expression-L4-R5,
L4Expression-S-C1-3Alkylidene-or-S (O)m,
R5Indicate hydrogen atom, C1-4Alkyl, amino ,-NH (C1-3Alkyl) ,-N (C1-3Alkyl)2,-OH ,-O (C1-3Alkyl) ,-C (O)(C1-3Alkyl) or 4-7 member naphthenic base,
The C1-4Moieties, naphthenic base, phenyl, heteroaryl, spirane structure and ring structure, caged scaffold can be into one Step is by 1 to 4 Q1Substitution,
Q1Indicate halogen atom, C1-3Alkyl, amino, C1-3Alkyl amino, two-(C1-3Alkyl) amino, hydroxyl, C1-3Alcoxyl Base, C1-3Alkoxy carbonyl group, carbamoyl, C1-3Alkyl-carbamoyl, two-(C1-3Alkyl) carbamoyl or 5-6 member cycloalkanes Base, wherein Q1It can be identical or different；
Carbon atom can be by 1-4 identical or different N, NH, N (C in the naphthenic base, bicyclic ring structures1-3Alkyl), O, S (O)m, C (O) replace；
The heteroaryl, spirane structure and ring structure, caged scaffold contain 1-4 hetero atom, be independently selected from N, O or S；
M indicates 0,1 or 2；
The expressions 0,1,2,3 or 4 of p and q independently.
In another preferred embodiment, the present invention provides above-mentioned logical formula (I) compound represented, its pharmaceutically Acceptable salt or its stereoisomer, wherein：
The expression phenyl of ring A and ring B independently contains the heteroatomic 5-6 membered heterocycloalkyls of N or 5-6 unit's heteroaryls；
L1And L2Expression-NH- ,-O- or-S (O) independentlym-；
A indicates covalent bond or imino group；
B expressions-CO-；
C indicates 1,2- ethenylidenes；
D indicates covalent bond or methylene；
E indicates hydrogen atom, piperidyl, morpholinyl, pyrrolidinyl, piperazinyl or two-(methyl) amino；
R1Indicate hydrogen atom, halogen atom, unsubstituted or the methyl, the methoxyl group that are replaced by one to three halogen atom, Methylamino or two-(methyl) amino；
R3Indicate hydrogen atom, halogen atom or-L3-R4,
L3Indicate covalent bond ,-NH- ,-N (C1-3Alkyl)-,-O- ,-O-C1-3Alkylidene-,-S-C1-3Alkylidene-,-S (O)m,-C (O)-,-NHC (O)-,-C (O) NH- ,-OC (O)-or-C (O) O-,
R4Indicate hydrogen atom, methyl, ethyl ,-N (C1-3Alkyl)2,-NHC (O) O-CH3,-O (CH3) ,-O (CH2CH3) ,-O (C(CH3)3) ,-S (O)2-CH3, pentamethylene base, cyclohexyl, pyrrolidinyl, tetrahydrofuran base, piperidyl, morpholinyl, piperazine Base, phenyl, pyrrole radicals, imidazole radicals, thiazolyl , oxazolyls, thiadiazolyl group, pyridyl group or pyrimidine radicals；
R2Indicate methylsulfany, ethylsulfanyl, methyl sulphonyl, amino-sulfonyl, methylsulfinyl or amino Asia sulphur Acyl group,
The methylsulfany, ethylsulfanyl, methyl sulphonyl, amino-sulfonyl, methylsulfinyl, amino sulfinyl It can be further by one or two identical or different methoxyl group, pyrrolidinyl, imidazolidine base, piperidyl, morpholinyl, piperazine Base replaces,
The pyrrolidinyl, imidazolidine base, piperidyl, pyridyl group, morpholinyl, piperazinyl, phenyl, pyrrole radicals, imidazoles Base, thiazolyl, oxazolyls, thiadiazolyl group, pyridyl group or pyrimidine radicals can be further by one or two identical or different Q1It takes Generation,
Q1Indicate halogen atom, methyl, amino, methylamino, two-(methyl) amino, hydroxyl, methoxyl group, methoxycarbonyl group, Carbamoyl, methylcarbamoyl or two-(methyl) carbamoyls；
M indicates 0,1 or 2；
The expressions 0,1 or 2 of p and q independently.
In another preferred embodiment, the present invention provides above-mentioned logical formula (I) compound represented, its pharmaceutically Acceptable salt or its stereoisomer, wherein：
The expression phenyl of ring A and ring B independently, pyridyl group, piperidyl, imidazole radicals, pyrazolyl, thiazolyl, thienyl, 1,2,3-triazoles base or 1,2,4- triazolyls；
L1And L2Expression-NH- independently or-O-；
A indicates covalent bond or imino group；
B expressions-CO-；
C indicates 1,2- ethenylidenes；
D indicates covalent bond or methylene；
E indicates hydrogen atom, piperidyl, pyrrolidinyl, morpholinyl, piperazinyl or two-(methyl) amino；
R1Indicate hydrogen atom, fluorine atom, chlorine atom, trifluoromethyl, methoxyl group or trifluoromethoxy；
R3Indicate hydrogen atom, fluorine atom, chlorine atom or-L3-R4,
L3Indicate covalent bond ,-NH- ,-N (C1-3Alkyl)-,-O- ,-O-CH2CH2,-S-CH2CH2Or-S (O)m,
R4Indicate hydrogen atom, methyl, ethyl ,-N (C1-3Alkyl)2,-NHC (O) O-CH3,-O (CH3) ,-O (C (CH3)3) ,-S (O)2-CH3, pyrrolidinyl, tetrahydrofuran base, piperidyl, morpholinyl, phenyl, pyrrole radicals, imidazole radicals, thiazolyl , oxazolyls, thiophene Di azoly or pyridyl group,
The phenyl, pyrrole radicals, imidazole radicals, thiazolyl, oxazolyls, thiadiazolyl group, pyridyl group can further by one to Two identical or different Q1Substitution,
Q1Indicate carbamoyl, methylcarbamoyl or two-(methyl) carbamoyls；
R2Indicate ethylsulfanyl that is unsubstituted or being replaced by methoxyl group, piperidyl, morpholinyl, methylsulfany, methyl sulphur Acyl group, amino-sulfonyl, methylsulfinyl, amino sulfinyl；
M indicates 0,1 or 2；
The expressions 0 or 1 of p and q independently.
In another preferred embodiment, the present invention provides above-mentioned logical formula (I) compound represented, its pharmaceutically Acceptable salt or its stereoisomer, wherein：
The expression phenyl of ring A and ring B independently, pyridyl group, pyrazolyl or piperidyl；
L1And L2Expression-NH- independently；
A indicates imino group；
B expressions-CO-；
C indicates 1,2- ethenylidenes；
D indicates covalent bond or methylene；
E indicates hydrogen atom, pyrrolidinyl, morpholinyl, piperazinyl or two-(methyl) amino；
R1Indicate hydrogen atom；
R3Expression-L3-R4, L3Indicate covalent bond ,-O- or-O-CH2CH2, R4Expression-N (CH3)2,-O (CH3), tetrahydrochysene furan It mutters base, morpholinyl, phenyl or pyridyl group,
The phenyl, pyridyl group can be further by one or two identical or different Q1Substitution, Q1Indicate carbamyl Base, methylcarbamoyl or two-(methyl) carbamoyls；
R2Indicate ethylsulfanyl that is unsubstituted or being replaced by methoxyl group, morpholinyl, methylsulfany, methyl sulphonyl, ammonia Base sulfonyl, methylsulfinyl, amino sulfinyl；
P indicates 1；
Q indicates 1.
In another preferred embodiment, the present invention provides above-mentioned logical formula (I) compound represented, its pharmaceutically Acceptable salt or its stereoisomer, wherein：
Ring A indicates phenyl；Ring B indicates phenyl, pyrazolyl or pyridyl group；L1And L2Expression-NH- independently；
A indicates imino group；B expressions-CO-；C indicates 1,2- ethenylidenes；D indicates covalent bond；
E indicates hydrogen atom；R1Indicate hydrogen atom；
R3Expression-L3-R4, L3Indicate covalent bond ,-NH- ,-N (C1-3Alkyl)-,-O- ,-O-C1-3Alkylidene-,-S-C1-3It is sub- Alkyl-, or-C (O) NH-,
R4Indicate hydrogen atom, methyl, ethyl ,-O (CH3) ,-O (C (CH3)3) or morpholinyl；
R2Expression-L4-R5, L4Expression-S-C1-3Alkylidene-or-S (O)m, R5Indicate hydrogen atom or C1-4Alkyl；
P indicates 1；Q indicates 1.
Detailed description of the invention
" C of the present invention1-6Alkyl " refers to the alkyl of the linear chain or branched chain containing 1-6 carbon atom, including “C1-4Alkyl ", " C1-3Alkyl " etc., the example include but not limited to such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, 2- Methyl-propyl, 1- methyl-propyls, 1,1- dimethyl ethyls etc..Term " C1-4Alkyl ", " C1-3Alkyl " refers to containing in examples detailed above There is the specific example of 1 to 4,1 to 3 carbon atom.
" C of the present invention1-6Alkylidene " refers to that the alkyl of the linear chain or branched chain containing 1-6 carbon atom removes one Structure after hydrogen atom refers to divalent alkyl." alkylidene chain " is polymethylene, i.e. ,-(CH2) x-, wherein x is positive integer, excellent It is selected as 1 to 6,1 to 4,1 to 3,1 to 2 or 2 to 3.It is one or more methylene hydrogen atoms through taking to be substituted alkylidene chain For the polymethylene of base displacement.Including " C1-4Alkylidene ", " C1-3Alkylidene " etc., the example include but not limited to for example sub- Methyl (- CH2), ethylidene (- CH2CH2), propylidene (- CH2CH2CH2), butylidene (- CH2CH2CH2CH2) etc..Term “C1-4Alkylidene ", " C1-3Alkylidene " refers to the specific example containing 1 to 4,1 to 3 carbon atom in examples detailed above.
" C of the present invention2-4Alkenyl " refers to the linear chain or branched chain alkenyl that the carbon atom number containing double bond is 2-4；In fact Example includes but not limited to such as vinyl, 1- acrylic, 2- acrylic, 1- methyl ethylenes, 1- cyclobutenyls, 2- cyclobutenyls, 3- Cyclobutenyl, 1- methyl-1-propylenes base, 2- methyl-1-propylenes base, 1- methyl -2- acrylic, 2- methyl -2- acrylic.
" C of the present invention2-4Alkynyl " refers to the alkynyl for the linear chain or branched chain that the carbon atom number containing three key is 2-4；Its Example includes but not limited to such as acetenyl, 2-propynyl, 2- butynyls, 3- butynyls, 1- methyl -2-propynyl.
" C of the present invention1-4Alkoxy ", " C1-4Alkylamino ", " two (C1-4Alkyl) amino ", " C1-4Alkoxy carbonyl Base ", " C1-4Alkyl sulfenyl ", " C1-4Alkyl sulphonyl ", " C1-4Alkyl sulphinyl ", " C1-4Alkyl amino sulfonyl ", " C1-4 Alkyl amino sulfinyl ", refers respectively to " C1-4Alkyl-O- " groups, " C1-4Alkyl-NH- " groups, " (C1-4Alkyl)2N- " bases Group, " C1-4Alkyl-O-C (O)-" groups, " C1-4Alkyl-S- " groups, " C1-4Alkyl-SO2" group, " C1-4Alkyl-SO- " bases Group, " C1-4Alkyl-SO2-NH-”、“C1-4Alkyl-SO-NH- " groups, wherein " C1-4Alkyl " is as defined hereinabove.
" C of the present invention1-6Alkoxy " refers to term " C1-6The group that alkyl " is connected by oxygen atom with other structures, Such as methoxyl group, ethyoxyl, propoxyl group, isopropoxy, butoxy, isobutoxy, tert-butoxy, sec-butoxy, amoxy, new penta Oxygroup, hexyloxy etc..It is preferred that C1-4Alkoxy, more preferable C1-3Alkoxy.Term " C1-4Alkoxy ", " C1-3Alkoxy " refers to term “C1-4Alkyl ", " C1-3The group that alkyl " is connected by oxygen atom with other structures.
" C of the present invention1-6Alkyl amido " refers to " C1-6The base that alkyl " is connected by acylamino- with other structures Group.
" halogen " of the present invention refers to fluorine atom, chlorine atom, bromine atom or iodine atom etc..
" 3-7 members naphthenic base " of the present invention refers to all carbon atoms of annular atom, is removed derived from a hydrogen atom Cyclic alkyl radical, including such as " 3-6 members naphthenic base ", " 4-6 members naphthenic base " " 5-7 members naphthenic base ", " 5-6 member cycloalkanes Base ", the example include but not limited to：Cyclopropane base, cyclobutane base, pentamethylene base, cyclohexyl, cycloheptyl alkyl, cyclooctane base, Cyclopentanone base, hexamethylene ketone group etc..
" 3-7 membered heterocycloalkyls " of the present invention refer to containing one or more heteroatomic 3-7 cyclic group, it is described " hetero atom " refers to nitrogen-atoms, oxygen atom, sulphur atom etc..It is preferred that 3-6 circle heterocyclic ring bases, more preferable 5-6 circle heterocyclic rings base.Specific example 2,5- dihydrothiophenes, 4,5- pyrazolines base, 3,4- dihydro -2H- pyranoses, 5,6- dihydro -4H-1 are included but are not limited to, 3- oxazinyls, aziridine base, azetidinyl, aza-cyclopentane base, piperidyl, Thietane base, tetrahydrofuran Base, nafoxidine base, imidazolidinyl, pyrazolidinyl, tetrahydrofuran base, Isosorbide-5-Nitrae-dioxane base, 1,3- dioxa hexamethylenes Alkyl, 1,3- dithians base, morpholinyl, piperazinyl,Deng.
" 4-7 unit's heteroaryls " refers to the armaticity constituted containing 4-7 annular atom (hetero atom there are one wherein at least containing) Group, including " 5-7 unit's heteroaryls " " 5-6 unit's heteroaryls ", specific example includes but are not limited to furyl, thienyl, pyrrole Cough up base, thiazolyl, thiadiazolyl group, oxazolyl, oxadiazolyls, imidazole radicals, pyrazolyl, pyridyl group, pyrimidine radicals, Isosorbide-5-Nitrae-dioxane Hexadienyl, 2H-1,2- oxazinyls, 4H-1,2- oxazinyls, 6H-1,2- oxazinyls, 4H-1,3- oxazinyls, 6H-1,3- oxazines Base, 4H-1,4- oxazinyls, pyridazinyl, pyrazinyl, 1,2,3- triazine radicals, 1,2,4- triazine radicals, 1,3,5-triazines base, 1,2,4, 5- tetrazines base, oxepin base, thia cycloheptatriene base, azepine cycloheptatriene base, 1,3- diazas cycloheptatriene base, nitrogen Miscellaneous cyclooctatetraenyl etc..
" 6-10 members bicyclic ring structures ", which refer to the bicyclic group constituted containing 6-10 annular atom, (can not contain or contain One and more than one hetero atom), including " 7-10 members bicyclic ring structures ", " 8-10 members bicyclic ring structures ", " 6-9 member loop coil knots Structure ", " 6-8 members and ring structure ", " 6-8 members caged scaffold " etc..Its specific example includes but not limited to two rings [3.1.0] hexane Base, two rings [4.1.0] heptane base, two rings [2.2.0] hexyl, two rings [3.2.0] heptane base, two rings [4.2.0] octyl, eight It is hydrogen pentalene base, octahydro -1H- indenyls, decahydronaphthalene naphthalene, ten tetrahydrochysene phenanthryl, bicyclic [3.1.0] hex- 2- alkenyls, bicyclic [4.1.0] hept- 3- alkenyls, bicyclic [3.2.0] hept- 3- alkenyls, bicyclic [4.2.0] octyl- 3- alkenyls, 1,2,3,3a- tetrahydrochysenes and ring Pentadienyl, 2,3,3a, 4,7,7a- hexahydro -1H- indenyls, 1,2,3,4,4a, 5,6,8a- octahydro naphthalenes, 1,2,4a, 5,6, 8a- hexahydros naphthalene, 1,2,3,4,5,6,7,8,9,10- decahydro phenanthryl, cyclobutane and nafoxidine base, pentamethylene and tetrahydrochysene pyrrole Cough up base, azetidine and imidazolidinyl, 3- oxabicyclos simultaneously [3.1.0] hexyl, hexahydro furyl [3,4-b] [Isosorbide-5-Nitrae] bis- Evil Ying Ji, hexahydro -2H- pentamethylene simultaneously [b] [Isosorbide-5-Nitrae] bioxin base, And two rings of armaticity Structure, including but not limited to benzofuranyl, benzisoxa furyl, benzothienyl, indyl, benzoxazolyl, benzo miaow Oxazolyl, indazolyl, benzotriazole base, quinolyl, isoquinolyl, acridinyl, phenanthridinyl, benzo pyridazinyl, phthalazinyl, quinazoline Base, quinoxalinyl, phenol piperazine base, pteridine radicals, purine radicals, naphthyridines base, 1,3- dihydro benzo furyls, benzo [d] [1.3] dioxa Cyclopentenyl, isoindoline base, Chromanyl, 1,2,3,4- nafoxidines simultaneously [3,4-c] pyrrole radicals, 5,6- glyoxalidine [1.2- A] pyrazine -7 (8H)-base, -7 (8H)-base of 5,6- dihydros -1,7- naphthyridines, 5H- pyrroles [3.4-b] pyridine -6 (7H)-base, 7,8- bis- Pyridinium hydroxide [4.3-d] pyrimidine -6 (5H)-base, 2,3,6,7- tetrahydrochysene -1H- pyrazoles [4.3-c] pyridine -5 (4H)-bases, 6,7- dihydros Thiazole [5.4-c] pyridine -5 (4H)-base etc..
" 6-9 member loop coils base " of the present invention refers to that a kind of 6-9 members that the formation of an atom is at least shared there are two ring are thick Ring structure.Its specific embodiment includes but are not limited to：Spiral shell [3.3] heptane base, spiral shell [3.4] octyl, spiral shell [3.5] nonyl, Spiral shell [4.4] nonyl, spiral shell [3.4] oct-6-ene base, spiral shell [3.5] nonyl- 6- alkenyls, spiral shell [4.4] nonyl- 6- alkenyls, spiral shell [4.4] nonyl- 2, 7- dialkylenes, 2- oxa- spiroheptanes base, 6- oxaspiros [2.5] octyl, 4- oxa-s -7- amino spiral shell [2.5] octyl, 2- amino spiral shell [3.3] heptane base, 2- oxa-s -6- amino spiral shell [3.3] heptane base, 2- amino spiral shell [3.4] octyl, 6- oxa-s -2- Amino spiral shell [3.4] octyl, 2- oxa-s -6- amino spiral shell [3.4] octyl, 2- oxaspiros [3.4] octyl, 5- oxaspiros [3.5] nonyl, 7- amino spiral shell [3.5] nonyl, 2- amino spiral shell [4.4] nonyl, 2- oxa-s -7- amino spiral shell [4.4] nonane Base, 2- oxaspiros [4.4] nonyl, 1,7- dioxo spiros [4.4] nonyl, Isosorbide-5-Nitrae, 7- trioxas spiral shell [4.4] nonyl, 6- ammonia Base spiral shell [3.4] octyl- 7- alkenyls, 2- oxa-s -6- amino spiral shell [3.4] octyl- 7- alkenyls, 7- amino spiral shell [3.5] nonyl- 5- alkenyls, 2- ammonia Base spiral shell [4.4] nonyl- 7- alkenyls etc..
" 6-8 member and ring structure " of the present invention refer to shared each other by two or more cyclic structures two it is adjacent Atom be formed by 6-8 cyclic groups, specific embodiment includes but are not limited to：Two rings [3.1.0] hexyl, two rings It is [4.1.0] heptane base, two rings [2.2.0] hexyl, two rings [3.2.0] heptane base, two rings [4.2.0] octyl, bicyclic [3.1.0] hex- 2- alkenyls, bicyclic [4.1.0] hept- 3- alkenyls, bicyclic [3.2.0] hept- 3- alkenyls, bicyclic [4.2.0] octyl- 3- alkene Base, benzofuranyl, benzisoxa furyl, benzothienyl, indyl, benzoxazolyl, benzimidazolyl, indazolyl, benzene And triazolyl, quinolyl, isoquinolyl, acridinyl, phenanthridinyl, benzo pyridazinyl, phthalazinyl, quinazolyl, quinoxalinyl, thiophene Pheno simultaneously [2,3-b] thienyl, thieno [3,2-b] thienyl, benzo [b] thienyl, benzo [b] thiazolyl, cyclobutane and four Hydrogen pyrrole radicals, pentamethylene and nafoxidine base, azetidine and imidazolidinyl, 3- oxabicyclos simultaneously [3.1.0] hexyl, six Hydrogen furans [3,4-b] [Isosorbide-5-Nitrae] bioxin base, hexahydro -2H- pentamethylene simultaneously [b] [Isosorbide-5-Nitrae] bioxin bases etc..
" 6-8 members caged scaffold " of the present invention refers to sharing two not phases each other by two or more cyclic structures Adjacent atom is formed by 6-8 cyclic groups, and specific embodiment includes but are not limited to： Deng.
Group described above can be further by 1 to 4 Q1Substitution, Q1Indicate halogen atom, C1-3Alkyl, amino, C1-3Alkane Base amino, two-(C1-3Alkyl) amino, hydroxyl, C1-3Alkoxy, C1-3Alkoxy carbonyl group, carbamoyl, C1-3Alkyl carbamoyl Base, two-(C1-3Alkyl) carbamoyl or 3-6 member naphthenic base, wherein Q1It can be identical or different.
Logical formula (I) compound represented specifically preferred according to the invention includes：
Its pharmaceutically acceptable salt or its stereoisomer.
Logical formula (I) compound represented specifically preferred according to the invention further includes：
Its pharmaceutically acceptable salt or its stereoisomer.
In one embodiment, the present invention provides the preparation methods of aforementioned present invention compound.
In one embodiment, the present invention provides the pharmaceutical compositions containing aforementioned present invention compound.
In one embodiment, the present invention provides use aforementioned present invention compound to prevent and/or treat B in individual The relevant leukemia of cell (such as B cell chronic lymphocytic cancer, non-Hodgkin lymphoma), inflammatory and autoimmune disease The method of (such as rheumatoid arthritis, systemic loupus erythematosus etc.).
In one embodiment, the present invention provides aforementioned present invention compounds to prepare for preventing and/or treating B The relevant leukemia of cell (such as B cell chronic lymphocytic cancer, non-Hodgkin lymphoma), inflammatory and autoimmune disease Application in the drug of (such as rheumatoid arthritis, systemic loupus erythematosus etc.).
Method described in following flows and/or those of ordinary skill in the art have may be used in above compound of the present invention Other technologies for knowing synthesize, but are not limited only to following methods.
For convenience, the present invention represents a variety of chemical compounds using well-known abbreviation, including but not limited to
DMF：N,N-Dimethylformamide；THF：Tetrahydrofuran；DIEA：N,N-diisopropylethylamine；
BINAP：2,2 '-bis- diphenyl phosphines-, 1,1 '-dinaphthalene；Xantphos：4,5- bis- diphenyl phosphine -9,9- dimethyl oxa-s Anthracene etc..
Reaction route：
Reaction step：
The preparation of step 1 intermediate 1
5-bromouracil (1 equivalent) is mixed with raw material 1 (at least 5 equivalents), it is solvent-free, or a small amount of polar solvent is added (water, ethyl alcohol, n-butanol etc.) heating (100-150 DEG C) is stirred to react a few hours to 5-bromouracil and disappears.It is cooling, it is precipitated solid Body, filtering, obtains intermediate 1.When raw material 1 is mercaptan, equivalent alkali (such as sodium hydroxide) need to be added in reaction.
The preparation of step 2 intermediate 2
Intermediate 1 (1 equivalent) is mixed with quantity of solvent phosphorus oxychloride (at least 10 equivalents), about two equivalent N, N- diformazans are added Base aniline, heating (90-110 DEG C) are stirred to react a few hours to intermediate 1 and disappear.It is cooling, it pours into ice water, solid, mistake is precipitated Filter, it is dry, obtain intermediate 2.Or concentration reaction system, column chromatography obtain intermediate 2.
The preparation of step 3 intermediate 3
Method 1：Intermediate 2 (1 equivalent) is mixed with raw material 2 (1.1-1.3 equivalents), appropriate solvent (such as tetrahydrochysene furan is added It mutters, n-butanol etc.), heating (80-120 DEG C) is stirred to react a few hours to intermediate 2 and disappears.It is cooling, concentrate reaction system, column layer Analysis, obtains intermediate 3.
Method 2：It is coupled using Buchwald-Hartwig.Intermediate 2 (1 equivalent) and raw material 2 (1-1.3 equivalents) is mixed It closes, appropriate solvent (being usually toluene, dioxane etc.) is added, the palladium catalyst of catalytic amount (is usually Pd2(dba)3, Pd (OAc)2Deng), the ligand (being usually Xantphos, BINAP etc.) of catalytic amount, alkali (Cs2CO3, NaOtBu etc.) and (1.1-1.5 works as Amount), it is sufficiently displaced from inert gas (such as nitrogen, argon gas etc.), heating (70-120 DEG C) is stirred to react a few hours to intermediate 2 and disappears It loses.It is cooling, reaction system is concentrated, column chromatography obtains intermediate 3.
The preparation of step 4 intermediate 4
Method 1：Intermediate 3 (1 equivalent) is mixed with raw material 3 (1-1.3 equivalents), appropriate solvent (such as tert-butyl alcohol, uncle is added Amylalcohol etc.), the acid (such as acetic acid, trifluoroacetic acid etc.) of alkali (such as DIEA, 1-1.5 equivalent) or catalytic amount is added, heats (80- 120 DEG C) or microwave under heating (80-120 DEG C) be stirred to react a few hours to intermediate 3 disappear.It is cooling, reaction system is concentrated, Column chromatography obtains intermediate 4.
Method 2：It is coupled using Buchwald-Hartwig.The method 2 with step 3 is operated, need to only change raw material 2 into raw material 3。
The preparation of step 5 intermediate 5
Intermediate 4 (1 equivalent) is dissolved in dichloromethane, quantity of solvent trifluoroacetic acid is added, or be passed through hydrogen chloride gas, room temperature or It is stirred to react under cooling to intermediate 4 and is disappeared.Concentration, obtains intermediate 5, is directly used in the next step.
The preparation of step 6 formula (I) compound
By intermediate 5 (1 equivalent) be dissolved in solvent appropriate (such as THF, dichloromethane, acetone, DMF, or for mix it is molten Agent), the alkali (such as DIEA) of 2-3 equivalents is added, under cooling (- 20 DEG C to -10 degree DEG C), is slowly dropped into raw material 4 (0.9-1.1 equivalents) It is stirred to react to intermediate 5 and disappears.Reaction is quenched, concentrates, column chromatography or mesolow prepare liquid phase and purify to obtain compound of formula I.
R in reaction route1、R2、R3、L1、L2, a, b, c, d, e, p, q, A and B it is as mentioned before.
In addition, above compound of the present invention or intermediate can also be synthesized using the method described in following flows, But it is not limited only to following methods：
Method 1：Substrate (1 equivalent) is dissolved in first alcohol and water (or in the mixed solvent of dichloromethane, methanol, water), Oxidizer (such as ammonium persulfate-sodium bisulfate, metachloroperbenzoic acid) (1 equivalent) reacts at normal temperatures to LCMS and monitors bottom Object completely disappears, and solution is spin-dried for, and prepares liquid phase purifying or silica gel column purification obtains product.
Method 2：Substrate (1 equivalent) is dissolved in methanol, a few hours of flowing back is open in air, is spin-dried for, prepares liquid-phase pure Change or silica gel column purification obtains product.
Substrate (1 equivalent) is dissolved in first alcohol and water (or in the mixed solvent of dichloromethane, methanol, water), oxygenation Agent (such as ammonium persulfate-sodium bisulfate, metachloroperbenzoic acid) (is more than 2 equivalents), and reaction at normal temperatures is stayed overnight or heating number Hour, until LCMS monitoring substrates completely disappear, solution is spin-dried for, liquid phase purifying is prepared or silica gel column purification obtains product.
R in reaction route1、R2、R3、R5、L1、L2, a, b, c, d, e, p, q, A and B it is as mentioned before.
Formula (I) compound of the present invention, its stereoisomer can in a free form or its pharmaceutically acceptable salt Form uses.The aobvious alkalinity of formula (I) compound of the present invention can form acid salt with inorganic acid or organic acid.Such as hydrochloride, hydrogen fluorine Hydrochlorate, hydrobromate, hydriodate, sulfate, trifluoroacetate, benzene sulfonate, mesylate, fluoroform sulphonate, second sulphur Hydrochlorate, carbonate, nitrate, phosphate, phosphite, maleate, tartrate, citrate, acetate, benzoic acid Salt, fumarate, oxalates, gluconate, hydroxyl acetate, isethionate, lactate, Lactobionate, malate, Succinate, tosilate, glycinate, trimethylglycine salt, arginine salt, ornithine salt, glutamate, asparagus fern Propylhomoserin salt etc..
Formula (I) compound of the present invention or its pharmaceutically acceptable salt, can be with one kind since there are asymmetric carbon atoms Optical isomeric form exists, and therefore, the invention also includes these optical isomers and its mixtures.Structure described herein Quasi- includes all isomeries (such as enantiomerism, diastereo-isomerism and geometrical isomerism (or conformational isomerism)) form of the structure； For example, R and S configurations, Z and E double bond isomers and Z and E rotamers about each asymmetric center.Therefore, this hair (or conformation is different for the single three-dimensional chemical isomer and enantiomter of bright compound, diastereoisomer and geometric isomer Structure body) mixture be within.Unless otherwise designated, otherwise all tautomerism shapes of the compounds of this invention Formula is within.In addition, unless otherwise designated, otherwise structure as described herein is also intended to include that difference only exists In there are the compounds of the atom of one or more isotope enrichments.For example, there is the structure of the present invention but include The compound of carbon displacement of the hydrogen through deuterium or tritium displacement or carbon enriched 13C or 14C is within the scope of the invention.Such compound can Therapeutic agent as probe or the present invention in such as analysis tool, bioanalysis.In some embodiments, include in Formulas I One or more D-atoms.
Formula (I) compound, its pharmaceutically acceptable salt or its stereoisomer of the present invention can be with one or more medicines With vehicle group at pharmaceutical composition.The conventional formulation clinically used can be made in described pharmaceutical composition, can take orally or intestines The modes such as stomach external administration are applied to the patient for needing this treatment.Such as tablet, particle, capsule, powder, injection, inhalant, tongue Lower drug-delivery preparation, syrup, gel, ointment, suppository, lotion, eye drops, nasal cavity drop, spray, preparation capable of permeating skin etc..These Preparation can by conventional method, add pharmaceutical carrier such as excipient, binder, humidizer, disintegrant, thickener etc. prepare and At.
Formula (I) compound, its pharmaceutically acceptable salt or its stereoisomer of the present invention press down with preferable BTK kinases It makes and uses, be that preferably there is excellent antitumor action and the drug for the treatment of autoimmune diseases effect.Formula simultaneously (I) compound, its pharmaceutically acceptable salt or its stereoisomer are preparing treatment B cell relevant leukemia (such as B cell Chronic lymphocytic cancer, non-Hodgkin lymphoma) and autoimmune disease (such as rheumatoid arthritis, systemic red Yabbi sore etc.) in play an important role.
Formula (I) compound, its pharmaceutically acceptable salt or its stereoisomer of the present invention are a kind of kinase inhibitors, special It is not Btk inhibitor.These inhibitor can be used for treating the disease of one or more response kinase inhibitions in mammal, Disease including response Btk inhibition and/or the inhibition of B cell proliferation.It is not intended to be bound by any specific theory, it is believed that this hair The interaction of bright compound and Btk lead to the inhibition of Btk activity, and therefore obtain these compound pharmaceutical applications.Therefore, originally Invention includes the mammal of the disease for treating the inhibition with response Btk activity and/or inhibition B cell proliferation, such as The method of people, this method include：A effective amount of at least one is administered to the mammal with such disease to carry herein The chemical entities of confession.Can be experimentally for example by the haemoconcentration of measurement compound, or theoretically pass through and calculate biology profit Expenditure determines effective concentration.Other than Btk, it is also possible to which affected other kinases include but not limited to other tyrosine Kinases and serine/threonine kinase.
Kinases rises in terms of the control elementary cell process such as signal transduction path of proliferation, differentiation and dead (Apoptosis) Obvious action.Abnormal kinase activity has implied that in various diseases, the disease includes kinds cancer, itself exempts from Epidemic disease and/or inflammatory disease and acute inflammatory response.Versatility effect of the kinases in key cells signal transduction path provides knowledge Not Ba Xiang kinases and signal transduction path novel drugs notable chance.
One embodiment includes treatment with autoimmunity and/or inflammatory disease or response Btk activity and/or B cell The method of the patient of the acute inflammatory response of the inhibition of proliferation.
The autoimmunity and/or inflammatory disease that can be influenced using compound according to the present invention and composition includes but not It is limited to：Psoriasis, allergy, regional enteritis, irritable bowel syndrome, sjogren disease, tissue graft rejection reaction and shifting The hyperacute rejection of organ is planted, asthma, systemic loupus erythematosus (and relevant glomerulonephritis), dermatomyositis is multiple Hardening, chorionitis, vasculitis (the relevant and other vasculitises of ANCA-), autoimmune haemolytic and thrombocytopenic disease Shape, Gourde(G) Paasche syndrome (and relevant glomerulonephritis and empsyxis), atherosclerosis, rheumatoid arthritis, slowly The Idiopathic Thrombocytopenic Purpura (ITP) of property, Addison disease, Parkinson's disease, Alzheimer disease, diabetes, septic Shock and myasthenia gravis.
Include herein is therapy, wherein at least one chemical entities provided herein are combined with anti-inflammatory agent Administration.Anti-inflammatory agent includes but not limited to：NSAID, non-specific and COX-2 specificity cyclooxygenase-2 inhibitors, gold compound, skin Matter steroid, methotrexate (MTX), tumor necrosis factor (TNF) receptor antagonist, immunosuppressor and methotrexate (MTX).
The example of NSAID includes but not limited to brufen, Flurbiprofen, naproxen and naproxen sodium, and Diclofenac is double The combination of chlorine fragrant sour sodium and Misoprostol, sulindac, benzene daybreak propionic acid, Diflunisal, piroxicam, Indomethacin, support degree Acid, fenoprofen calcium, Ketoprofen, Nabumetone sodium, sulfasalazine, tolmetin sodium and hydroxychloroquine.The example of NSAID further includes COX-2 specific inhibitors such as celecoxib, valdecoxib, Lu meter Kao former times and/or etoricoxib.
In some embodiments, anti-inflammatory agent is salicylate or salt.Salicylate or salt include but not limited to acetyl group Salicylic acid or aspirin, sodium salicylate and Choline Salicylate and magnesium salicylate.
Anti-inflammatory agent can also be cortical steroid.For example, cortical steroid can be cortisone, and dexamethasone, first Prednisolone, prednisolone, Inflamase or prednisone.
In a further embodiment, anti-inflammatory agent is gold compound, such as sodium aurothiomalate or Anranofin.
Metabolic poison such as dihydrofolate reductase inhibitor the invention also includes wherein anti-inflammatory agent, such as methotrexate (MTX) or The embodiment of dihydrooratic acid salt dehydrogenase inhibitor such as leflunomide.
It is anti-monoclonal antibody (such as according to library pearl that other embodiments of the present invention, which is related to wherein at least one anti-inflammatory compound, Monoclonal antibody or training gram pearl monoclonal antibody), the combination of TNF antagonists such as Etanercept (entanercept) or infliximab, the English profit Former times monoclonal antibody is a kind of anti-TNF alpha monoclonal antibody.
It is that immunosuppressant compounds are such as selected from that other embodiments of the present invention, which are related to wherein at least one active drug, Methotrexate (MTX), leflunomide, ring born of the same parents are plain, tacrolimus, the immunosuppressant compounds in imuran and mycophenolate mofetil Combination.
The B cell and B cell precursor for expressing Btk have implied that in the pernicious pathology of B cell, B cell is pernicious to include But it is not limited to B cell lymphoma, lymthoma (including Huo Qijin and non-Hodgkin lymphoma), hairy cell lymphoma, multiple bone Myeloma, chronic and acute myelogene leukemia and chronic and acute lymphocytic leukemia.
Have shown that Btk is Fas/APO-1 (CD-95) the death inducement signal conducting composite in B- systems lymphoid cell (DISC) inhibitor.The destiny of leukaemia/lymphoma cell may is that by the anti-of the DISC Caspases activated Cells apoptosis and including the balance between Btk and/or the upstream anti-apoptotic regulation mechanism of its substrate forward (Vassilev etc., J.Biol.Chem.1998,274,1646-1656).
It has also been found that Btk inhibitor may be used as chemical sensitizer, therefore can be used for combining with other chemotherapeutic drugs, institute The medicine of the chemotherapeutic drug stated especially inducing cell apoptosis, such as antitumor agent, immunosuppressor.It can press down with chemical sensitization The example for other chemotherapeutic drugs that formulation compositions use include but are not limited to topoisomerase I inhibitor (such as camptothecine or Hycamtin), Topoisomerase II inhibitors (such as daunomycin and Etoposide), alkylating agent (such as cyclophosphamide, melphalan and Medicament (such as taxol and vincaleukoblastinum) and biological agent that BCNU), tubulin is oriented to (such as antibody such as anti-CD 20 antibodies, IDEC8, immunotoxin and cell factor).
Btk activity is white with bcr-abl fusions caused by some transpositions of expression by chromosome dyad 9 and 22 Blood disease is related.This exception is usually observed in chronic myelogenous leukemia.Btk is substantially by bcr-abl kinases phosphorus Acidification, this initiation prevented in bcr-abl cells Apoptosis downstream survival signaling (N.Feldhahn etc., J.Exp.Med.2005,201 (11), 1837-1852).
The compounds of this invention has the following advantages compared with the immediate prior art：
(1) formula (I) compound of the present invention or its pharmaceutically acceptable salt have preferable BTK kinase inhibitory activities, peace Quan Xinggao, security window is big and Small side effects；
(2) formula (I) compound of the present invention or its pharmaceutically acceptable salt show good biological stability, when drug effect Journey is long, persistent, and bioavilability is high；
(3) the compounds of this invention preparation process is simple, and drug purity height, stable quality are easy to carry out large-scale industry life Production.
Below by way of pharmacological evaluation the present invention is further explained compound advantageous effect, but this should not be interpreted as to the present invention Compound only has following advantageous effect.
The pharmacological activity test of test example the compounds of this invention
External anti-bruton's tyrosine kinase (BTK) determination of activity of I the compounds of this invention
Test sample：
Control compound：CC-292 is made according to WO2009158571A1.
The compounds of this invention：Its chemical name and structural formula are shown in the preparation embodiment of each compound with preparation method.
Experimental method：
Representative meaning of abridging in following experiments is as follows：
ATP：Atriphos；BTK：Bruton's tyrosine kinase；mg：Milligram；mL：Milliliter；μg：Microgram；μL：Microlitre； mM：MM every liter；EDTA：Ethylenediamine tetra-acetic acid；DMSO：Dimethyl sulfoxide (DMSO).
1. test material
HTRFR KinEASETM-TK：Purchased from Cisbio, lot number 62TK0PEB；BTK：Purchased from Carna, Cat.No.08- 080；ATP：Purchased from Sigma, Cat.No.A7699, CAS No.34369-07-8；MgCl2：Purchased from Sigma, CAS No.7786- 30-3, Lot.No.101M8701V；DMSO：Purchased from Sigma, CAS No.67-68-5, Lot.No.STBC0365V；96 orifice plates： Purchased from Thermo, Cat.No.249944, Lot.No.1057825；384 orifice plates：Purchased from Greiner, Cat.No.784075, Lot.No.E1112Φ6Y。
2. experiment preparation of reagents
1 × kinase buffer liquid 1. (Kinase buffer) (5mM MgCl2, 1Mm DTT, 50nM SEB)；2. DTT is by DTT It is spare as storing solution that stoste uses sterilized water for injection to be diluted to 100mM；3. ATP prepares the deposit of 5mM with sterilized water for injection Liquid is spare；4. 10mM compound solutions：The compound stock solution that compound dissolving is configured to 10mM using 100%DMSO is standby With.
3. the enzyme reaction stage
1. by the compound solution of 10mM with 100%DMSO dilute 20 times, then carried out after diluting 2 times a series of 3 times it is dilute It releases, totally 10 concentration gradients, then uses 1 × kinase buffer that the solution of each concentration is diluted 100 times as experiment chemical combination Object concentration, 4 holes μ L/.2. preparing 5 × enzyme solutions：1 × kinase buffer, 2 holes μ L/ are added in enzyme.3. being incubated under the conditions of 25 DEG C Educate 30min.4. preparing 5 × TK Substrate-biotin：1 × kinase is added in TK Substrate-biotin In buffer, the holes 2mL/；5. preparing 5 × ATP：ATP is added in 1 × kinase base buffer, the holes 2mL/；6. at 25 DEG C Under the conditions of be incubated 40min.
4. detecting the stage of reaction
1. preparing 4 × Streptavidin-XL665：Streptavidin-XL665 is addedDetection In buffer, 5 holes μ L/.2. adding 5mL TK Antibody-Cryptate per hole.3. being incubated 60min under the conditions of 25 DEG C.
5. digital independent
After the completion of detecting the stage of reaction, fluorescent value of the detection sample at 615nm and 665nm is detected respectively with microplate reader.
6. curve matching obtains IC50
It is carried out curve fitting using 5.0 softwares of GraphPad, fit equation is Y=Bottom+ (Top-Bottom)/(1+ 10^ ((LogIC50-X) * HillSlope)), obtain IC50Value.
Experimental result：In said determination, the compounds of this invention and control compound CC-292 swash bruton's tyrosine The IC of the cell in vitro inhibitory activity of enzyme (BTK)50Value is listed in Table 1：
Table 1
Experiment conclusion：
In conclusion the compounds of this invention is significantly stronger than control compound CC-292 to the inhibitory activity of BTK kinases.
The external anti-cell determination of activity of II the compounds of this invention
The compounds of this invention：Its chemical name and structural formula are shown in preparation embodiment with preparation method.
1. laboratory apparatus
2104 plate readers of Envision, PerkinElmer, (U.S.)；CO2Incubator, SANYO. (Japan)；It is inverted aobvious Micro mirror, XDS-1B, Chongqing broadcasting and TV (Chongqing, China)；PH is counted, Mettler Toledo Five easy. (China)；MACS points From device (Miltenyi, the U.S.)；FACSCalibur (BD, the U.S.).
2. cell
In 37 DEG C, 5%CO2 incubators, with the fetal calf serum containing 10% ultralow immunoglobulin, penicillin/chain Mycin, 5mM HEPES, the RPMI1640 medium culture Balb/c mouse primary splenic B cells of 50 μM of beta -mercaptoethanols.Culture Base is purchased from U.S. GIBCO.
3. mouse
Male, 6-8 weeks Balb/c mouse.
4. reagent is prepared with compound
(1) CellTiter-Glo (CTG) (article No.：G7572, Promega), CTG buffer solutions and CTG substrates are stored in- 20 DEG C, recommend to prepare CTG reagents with the following method：
It using preceding thawing CTG buffer solutions and balances to room temperature, for the sake of convenient, CTG buffer solutions can be melted well simultaneously using preceding It is stored at room temperature by 48 hours or more.The CTG substrates of freeze-drying are equilibrated into room temperature, take the buffer solution of 100ml to equipped with substrate In amber bottle, CTG reagents have just been obtained.Gently mixing should be in one minute completely until obtaining uniform solution, substrate Melt, dispense and preserves CTG reagents for a long time in -20 DEG C of refrigerators.
(2)B cell separating kit (the number of ordering：130-090-862, Miltenyi)
(3) PE antibiotins (anti-biotin) antibody (article No.：409003, Biolegend)
(4) PE/cy7 exempts from anti-mouse CD45R/B220 antibody (article No.s：103221, Biolegend)
(5) cell strain (article No.：352340, BD Falcon)
(6)AffiniPure F(ab′)2Fragment goat anti mouse IgM, μ chain specific (goods Number：115-006-020, Jackson)
(7) test compound is prepared
·Prepare test Compound Stock solution：Compound powder is dissolved in DMSO, 10mM concentration is arrived.
·Prepare test compound gradient dilution solution：First, the test Compound Stock solution DMSO 3 of 10mM is taken to connect again Continue gradient dilution, totally 10 concentration.The diluted compounds of DMSO of 10 μ L are taken to be added to 90 μ L Compound Dilution Buffers respectively again In, then the diluted compound containing 10%DMSO of 10 μ L is taken to be added in 90 μ L Compound Dilution Buffers respectively, compound highest A concentration of 10 μM, DMSO a concentration of 0.1%, totally 10 concentration gradients.
5. experimental method
(1) Balb/c mouse B cells are detached：
Balb/c mouse spleens are taken, are smashed to pieces in MACS buffer solutions, list is obtained with 40 μm of Nylon cell screen filtration Cell suspension.
4 DEG C, obtained cell suspension is centrifuged five minutes in 400g, removes supernatant, the red blood cell that 1ml is added at room temperature is split Simultaneously cell mass has lightly been resuspended in solution liquid.After two minutes, the MACS buffer solutions of precooling are added.It is thin with 40 μm of cell screen clothes filtering In born of the same parents' suspension to a new centrifuge tube.4 DEG C, 400g centrifuges 5 minutes to collect cell.
Before magnetic bead is added, cell density is adjusted to 10 with MACS buffer solutions7The μ l of a cell/40.Every 107A cell The middle biotinylated antibody mixture that 10 μ l are added.It is incubated on ice after mixing 20 minutes, every 10730 μ l are added in a cell MACS buffer solutions and 20 μ l antibiotin magnetic bead and on ice be incubated 20 minutes.It is buffered with the MACS of 500 μ l after centrifugation Cell is resuspended in liquid.The MACS sorting columns of precooling are placed in MACS sorters, cell suspension is added in MACS sorting columns.It collects The cell of the unbonded antibody flowed down.
By flow cytometry PE anti-biotin antibody and CD45R (B220) antibody test detection sorting before and Cell after sorting.
(2)IC50It measures
The mouse B cell that fresh separated is counted with blood counting chamber expects that blue decoration method detects Cell viability and answers by a word used in place name 98% or more.
Cell density is adjusted to every milliliter 3.89 × 10 with culture medium5A cell takes 90 μ l thin with multichannel pipettor In born of the same parents' suspension to 96 orifice plates, it is 3.5 × 10 to obtain final cell density4A cell per well.
Storing liquid is formed with DMSO dissolved dilutions test compound and positive compound, a series of of 10 μ l preparations are added In compound solution to 96 orifice plates (each concentration of each compound does 3 points of repetitions).At 37 DEG C, 5%CO2It is incubated in incubator It educates 30 minutes, then adds 50 μ l B cells stimulation mixed liquor, it is 10 μ g/ to stimulate the final concentration of anti-Igm in mixed liquor ml。
By cell plates at 37 DEG C, 5%CO2Continue to be detected with the method for CTG after being incubated 72 hours in incubator.
Melt CTG reagents and balance to room temperature, is transferred in 96 orifice plates with multichannel pipettor, 50 μ l CTG reagents/hole, It is placed in the dark 10 minutes after being shaken 2 minutes on the quick oscillator of microwell plate, the readings that shines is detected with Envision.
6. data analysis
Obtained data can be analyzed with Excel 2007 and 5.0 softwares of GraphPad Prism, in order to calculate IC50, It will be returned using non-linear S curve and obtain a dose-effect curve, 5.0 software meetings of GraphPad Prism come fitting data Automatically IC is provided50Value.
Cell survival rate is calculated with following formula：VSample/V2Solvent control× 100%, VSampleIt is the reading in compound processing hole Value, V2Solvent controlIt is the average value of solvent control hole (V2) readings.
7. experimental result：
IC of the compounds of this invention (compound 1-106) to the inhibitory activity of Balb/c mouse B cell cell in vitro50＜ 0.5μM.The IC of part of compounds50It is as follows：
2 part of compounds of the present invention of table is to Balb/c mouse B cell cell in vitro inhibitory activity
Wherein, ++++represent IC50(μM)≤0.01μM；+++ represent 0.01 μM of ＜ IC50(μM)≤0.1μM；++ represent 0.1 ＜ IC50(μM)≤0.5μM。
Experiment conclusion：In conclusion as can be seen from Table 2, the compounds of this invention has relatively by force Balb/c mouse B cells proliferation Inhibiting effect.
III the compounds of this invention is to rat spleen cells BTK enzyme occupation rate determination experiments
In order to measure the amount for not occupied BTK in cell or tissue lysate by compound, ELISA schemes, profit are used The biotinylated probe compound of BTK is not occupied by compound with a kind of combination, evaluation compound is under various concentration, in spleen To the occupation rate situation of BTK enzymes in cell, %BTK occupation rates (BTK Occupancy) are calculated.
1, experiment material
The anti-BTK antibody of mouse (Becton Dickinson)；Goat anti-mouse HRP antibody (Becton Dickinson)； Cell pyrolysis liquid (Cell Signaling)；Bruton's tyrosine kinase (BTK) (Carna)；Coated 96 hole of Streptavidin Plate (Thermo)；(LTS 1083PK, the Tianjin oceans Hao biological products science and technology Limited Liability are public for rat lymphocyte separating kit Department)；Microplate reader (victor4, PE)；Centrifuge (5804R, Eppendorf)；Microscope (CX31RTSF, Olympus)；MACS Sorter (midiMACS separation unit, MACS).
2, experimental procedure
(1) preparation of reagents
Probe compound solution (Probe)：Weigh sample compound 1mg, compound concentration is the storing solution of 1mM, when use It is diluted with sample diluting liquid；Sample diluting liquid (Sample diluents)：Containing 1% bovine serum albumin(BSA) and 0.1%Tween-20 PBS；Cleaning solution (Washing solution)：PBS containing 0.05%Tween-20.
(2) the unicellular preparation of Rats Spleen
Spleen is rinsed with the PBS buffer solution of 1mL and (osculum is cut in one end of spleen, with syringe in the spleen other end 1ml precoolings PBS is injected to rinse), it is then transferred to the sterile strainer of 200 mesh, is shredded, then ground with syringe, is paid attention to operating scissors Add the PBS buffer solution of precooling to rinse in grinding, amounts to and rinsed with the PBS buffer solution of 5ml.4 DEG C, 400g centrifuges 3min, goes Clearly, 20ml cell washing solutions PBS is added, 4 DEG C, 600Rpm, centrifuges 10min, washs 3 times.
(3) compound and cytosis
After cell count, cell concentration is diluted to 3 × 107Cells/ml, 90 holes μ l/ with PBS.Compound 10 is added The holes μ l/, 37 DEG C of incubation 1h.20 DEG C, 400g centrifuges 20min, discards supernatant.
(4) lytic cell
Protease inhibitors PMSF is added in cell pyrolysis liquid (cell lysis buffer) and (notices that PMSF is being split It solves liquid and uses preceding addition).Mixing in the cell of often pipe enrichment is added in lysate, is operated according to lysate specification, is split on ice 5min is solved, 14000g centrifuges 10min.100 μ l of supernatant are taken to be added in 96 orifice plates.
(5) BTK determination experiments step
100ul standard items are added per hole or sample is mixed with 10 μ l probe compounds solution (final concentration of 1 μM), 28 DEG C of shakes It swings and is incubated 1h.After incubation, 100 μ l is taken to be added to (Streptavidin-coatd on the coated elisa plate of Streptavidin ELISA plate), 28 DEG C of concussions are incubated 1h.With cleaning solution (Washing sol ution) board-washing 5 times.It is added into every hole The anti-human BTK antibody of mouse (Purified mouse anti-human antibody) (1: 1000 times of dilution) of 100 μ l purifying.28 DEG C concussion be incubated 1h.With cleaning solution board-washing 5 times.Sheep anti-mouse antibody (the HRP goat of 100 μ l HRP labels are added into every hole Anti-mouse Ig) (1: 1000 dilution).28 DEG C of concussions are incubated 1h.With 5 backward 100 bottoms μ l of addition per hole of cleaning solution board-washing Object TMB solution, 28 DEG C of incubation 15min.50 μ l 1M H2SO4 are added per hole and terminate reaction.
3, Testing index and detection method
After reaction terminating, the OD values at 450nm are detected.According to OD values, calculated using 4 parameter logistic curves of A in microplate reader The amount of BTK in each sample.
4, data processing and result
* %BTK occupation rates=(control group BTK amounts-compound group BTK amounts)/control group BTK amount × 100%
The * coefficient of variation
Experimental result is as shown in table 3：
Table 3：Compound is to rat spleen cells BTK occupation rate experimental results
Experiment conclusion：By table 3 as it can be seen that BTK occupation rate of the compounds of this invention in rat spleen cells is higher, embody compared with Good drug effect.
IV the compounds of this invention plasma stabilities are tested
Experimental method
1, appraisement system
(1) compound and corresponding mother liquor information are tested：Compound 5mg or so is taken, the deposit of 10mM is first configured to DMSO Liquid, then with DMSO: water=1: 1 is diluted to the test compound working solution of 50uM, for use.(DMSO contents in reaction system≤ 0.5%)
(2) tested media
2. sample preparation methods
(1) by the blood plasma of frost set in 37 DEG C of thermostat water baths preincubate thaw it is spare.
(2) 10 μ L untested compounds (50 μM) are taken to be added in each kind blood plasma of 490 μ L, 1 μM of determinand final concentration, two multiple Sample.
(3) vortex mixing is placed in 37 DEG C of thermostat water baths and incubates.
(4) 50 μ L are extracted reaction solution at corresponding time point (T=0h, 2h, 4h), is added the terminate liquid of 300 μ L, after mixing - 80 DEG C are set to freeze.
(5) after the completion of waiting for incubation assays, thaw each time point sample cell, is centrifuged to 12000rpm in centrifuge after mixing 5min。
(6) it takes clean 96 hole sample panel that the water of 150 μ L is added, takes in 50 μ L to sample well of supernatant, sample is submitted after mixing Sample introduction is analyzed for plate progress.
Sample introduction is analyzed by 3.LC-MS/MS
4. data analysis
Stability of the compound in blood plasma is evaluated using reservation percentage of the compound after each time point is incubated, sample The ratio between peak area and internal standard peak area of the concentration test compound of product indicate.Calculation formula is as follows：
Surplus (%)=CTn/CT0
Wherein, CTnFor whole solution measured concentration of the compound after each incubation time point, n=2h, 4h；
CT0Whole solution measured concentration when being incubated is being originated for compound.
Experimental result
(T=2h) surplus (%) after different blood plasma medium temperatures are incubated of table 4 compares
(T=4h) surplus (%) after different blood plasma medium temperatures are incubated of table 5 compares
In conclusion the compounds of this invention 23, compared with comparison medicine CC-292, the stability in blood plasma is more preferable.
V patch clamp methods detect the inhibiting effect to hERG potassium-channels of the compounds of this invention
Experiment material：As follows：
* Amitriptyline Hydrochloride, international standard hERG channel blockers.
The compounds of this invention (compound 1-106)：Its chemical name and structural formula are shown in the system of each compound with preparation method Standby embodiment.Control compound：CC-292 is synthesized according to patent WO2009158571A1.
Experimental method
(1) preparation of solution and compound
Extracellular fluid (mM)：N-2- hydroxyethylpiperazins-N ' -2-ethanesulfonic acid (HEPES) 10, NaCl145, KCl 4, CaCl22、MgCl21,10 Glucose adjust pH to 7.4 with 1N sodium hydroxides；Osmotic pressure is adjusted to 290-300mOsm；4 after filtering DEG C preserve.
Liquid (in mM) in electrode：KCl 120、KOH31.25、CaCl25.374、MgCl21.75, (the beta-amino of ethylene glycol-two Ethylether)-N, N, N ', N '-tetraacethyls (EGTA) 10, HEPES 10, Na2- ATP 4 adjusts pH to 7.2 with 1N potassium hydroxide； Osmotic pressure is adjusted to 280-290mOsm；- 20 DEG C of preservations after filtering.
The preparation of compound：Positive control drug Amitriptyline Hydrochloride and 2 sample CC-292 and the compounds of this invention (chemical combination Object 1-106) it is first dissolved in 100%DMSO (Sigma-Aldrich, 34869), it is configured to the stock solution of 30mM.It is used before experiment Above-mentioned stock solution is diluted to the solution of 1000 times of each experimental concentration by DMSO, and then diluting 1000 times with extracellular fluid again arrives Required concentration.DMSO ultimate densities are 0.1% in extracellular fluid.
(2) electro physiology is tested.
HERG electric currents are recorded using whole-cell patch-clamp recording technique.It takes cell suspension to be added in the culture dish of 35mm, is placed in down It sets on microscope carrier.After cell is adherent, with extracellular fluid perfusion, flow velocity 1-2mL/min.Glass microelectrode is by micro- electricity Pole draws two step of instrument and draws, and it is 2-5M Ω to enter water power resistance value.After establishing whole-cell recording technique, holding command potential is -80mV.It gives It is depolarized to+60mV when giving voltage stimulation, then repolarization to -50mV draws hERG tail currents.All records are steady in electric current It is carried out after fixed.Since low concentration, each concentration 5-10min to electric current stablizes, then gives next concentration for extracellular perfusion administration.
(3) cell strain
Stable cell line CHO-hERG is purchased from AVIVA companies.For quality control, minimum sealing-in resistance is not less than 1G Ω, and hERG electric currents are not less than 0.4nA.
This time experiment includes the following aspects：
HERG electric currents are recorded on the CHO-K1 cell strains for stablizing the channels expression hERG using manual patch clamp technique；According to HERG tail currents calculate the inhibiting rate of each concentration；Each compound tests 5 concentration, calculates IC50Value；Each concentration determination 2 A cell；One positive control medicine.
(4) data acquisition and processing (DAP)
By Digidara 1440 (Molecular Devices) and pCLAMP softwares (10.2 editions, Molecular Devices) A/D-D/A digital-to-analogue conversions carry out stimulation granting and signal acquisition；Patch-clamp amplification (Multiclamp 700B, Molecular Devices) amplified signal, it is filtered into 1KHz.It uses Clampfit (10.2 editions, Molecular Devices) Data analysis and curve matching are further carried out with Prism.Data are indicated with means standard deviation.IC50Numerical value by Logistic equations are fitted gained：
y：Suppression percentage；max：It is 100%；min：It is 0%；[drug]：Tester concentration；nH：Slope；IC50：Test The maximum half-inhibition concentration of object.
Experimental result
This experiment utilizes whole-cell patch-clamp recording technique, the having detected on the CHO-Kl cell strains for stablizing the channels expression hERG Close blocking effect of the object to hERG electric currents.Half-inhibition concentration (the IC of each test compound50) best by Logistic equations Fitting obtains.The drug concentration that we select within the scope of the detectable concentration of this experiment ranging from institute of International Pharmaceutical research and development company is most Common hERG lane testing ranges, because this concentration range has covered common dosage.The overwhelming majority is to hERG Channel has the sample of inhibiting effect that can generate inhibiting effect to hERG electric currents at these concentrations.Resistance of the compound to hERG Disconnected effect is shown in Table 5.
The IC for the compounds on hERG currents that table 6 is recorded on CHO-K1 stable cell lines50Numerical value
Positive control medicine Amitriptyline is one of the most widely used blocking hERG electric current tool drugs, The IC that hERG electric currents are inhibited in this research50It it is 3.15 μM, this result is consistent with the result of document report.This shows this Secondary experiment the result is that believable.
The compounds on hERG currents of this research institute detection inhibits in highest test concentrations (30.00 μM) to hERG electric currents Inhibiting effect reaches far away IC50, to illustrate that the compounds of this invention (compound 1-106) is not apparent to the channels hERG Inhibiting effect.And comparison medicine CC-292 is to the IC of hERG electric currents50Value is 21.50 μM, has inhibiting effect to the channels hERG.Therefore, The compounds of this invention (compound 1-106) is compared with comparison medicine, safety higher.
Bibliography：Blockade of the HERG human cardiac K+channel by the Antidepressant drug amitriptyline.British Journal of Pharmacology, (2000) 129： 1474-1480.
The safety evaluatio of VI the compounds of this invention is tested
The compounds of this invention (compound 1-106)：Its chemical name and structural formula are shown in the system of each compound with preparation method Standby embodiment.
Experimental method
Observation SD rat oral gavages give test-compound (compound 1-106) repeat administration 14 days toxicity, understand tested Object toxic effect and its target organ determine nontoxic crude protein under this experimental condition；And whether tested material is observed by convalescence There are delayed toxicity and the toxic reaction of appearance whether reversible.
Master trip group is respectively vehicle control group, tested material (compound 1-106) low dose group, middle dose group, high dose Group, each 10 of animal, half male and half female；Wherein middle dose group and vehicle control group separately set each 10 of convalescence animal, half male and half female.
By 10ml/kg capacity gastric infusions, 1 time a day, successive administration 14 days.The appearance of animal is observed during experiment daily Change with general behavior, and carries out weight, food ration, hematology, coagulation function, serum biochemistry inspection；After administration phase Gross anatomy is carried out to all euthanasia animals, main organs are weighed, and internal organs carry out histopathologic examination.
Experimental result
The tolerance of the compounds of this invention (compound 1-106) is good, safe.
4, specific implementation mode
The specific implementation mode of form by the following examples makees further specifically the above of the present invention It is bright.But the range that this should not be interpreted as to the above-mentioned theme of the present invention is only limitted to following embodiment.It is all to be based on the above of the present invention The technology realized all belongs to the scope of the present invention.
1 4- of embodiment (4- (4- (3- acrylamidos phenylamino)-5- (methyl mercapto) pyrimidine -2 --amino) phenoxy group)- The preparation of N- picoline -2- formamides (compound 1)
(1) preparation of 5- (methyl mercapto) pyrimidine -2,4 (1H, 3H)-diketone
The aqueous solution (10mL) of 5-bromouracil (3g, 15.7mmol) and sodium methyl mercaptide 20% is mixed, is heated to 100 DEG C 5h is reacted, is cooled to room temperature, filters, obtains solid 1.5g, yield：60.5%.
The preparation of (2) 2,4- bis- chloro- 5- (methyl mercapto) pyrimidine
By 5- (methyl mercapto) pyrimidine -2,4 (1H, 3H)-diketone (1.5g, 9.5mmol), n,N-Dimethylaniline (2.3g, It 19mmol) mixes, phosphorus oxychloride (15g, 97.8mmol) is added, 4h is reacted at 100 DEG C, it is cooling, (100mL) is poured into ice water, Solid is precipitated, filters, it is dry, obtain solid 1.1g, yield：59.4%.
(3) preparation of 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate
By 2,4- bis- chloro- 5- (methyl mercapto) pyrimidine (0.5g, 2.56mmol), 3- aminophenyiamino t-butyl formates (0.68g, 3.26mmol) is dissolved in THF (20mL), and DIPEA (0.42g, 3.25mmol) is added, is heated to reflux 7h, is cooled to room temperature, Concentration, column chromatography (PE: EA=4: 1) obtain product 0.4g, yield：43%.
(4) 3- (2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) -5- (methyl mercapto) pyrimidines -4- Base amino) phenylcarbamate preparation
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.3g, 0.82mmol), 4- (4- amino-benzene oxygens)-N- picoline -2- formamides (0.24g, 0.99mmol) are dissolved in the tert-butyl alcohol (5mL), and DIPEA is added (0.127g, 0.98mmol) is heated to 90 DEG C of reaction 8h, is cooled to room temperature, filters, dry, obtains solid 0.4g, yield：85%.
(5) 4- (4- (4- (3- amino phenyl aminos)-5- (methyl mercapto) pyrimidine -2 --amino) phenoxy group)-N- picolines- The preparation of 2- formamides
By 3- (2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) -5- (methyl mercapto) pyrimidines -4- Base amino) phenylcarbamate (0.4g, 0.697mmol) is dissolved in DCM (10mL), CF is added3COOH (2mL) room temperature is stirred 2h is mixed, is spin-dried for, the next step is directly used in.
(6) 4- (4- (4- (3- acrylamidos phenylamino)-5- (methyl mercapto) pyrimidine -2 --amino) phenoxy group)-N- first The preparation of yl pyridines -2- formamides
4- (4- (4- (3- amino phenyl aminos)-5- (methyl mercapto) pyrimidine -2 --amino) phenoxy group)-N- that upper step is obtained Picoline -2- formamides, DIPEA (0.18g, 1.39mmol) are dissolved in THF (5mL), are cooled to -20 DEG C, in 20min slowly The 2mL THF solutions of acryloyl chloride (0.63g 0.696mmol) are added dropwise into system.It directly concentrates, prepares chromatogram purification (H2O∶ MeOH=100: 50) product white solid 90mg, two step yields 24.5% are obtained.
Molecular formula：C27H25N7O3S molecular weight：527.17 mass spectrum (m/z)：528.2(M+H)+.
1H-NMR(d6- DMSO, 400MHz) δ (ppm)：10.12 (1H, s), 9.51 (1H, s), 8.80-8.72 (2H, m), 8.47 (1H, d), 8.23 (1H, s), 7.87 (1H, m), 7.75 (2H, d), 7.42 (1H, d), 7.37-7.26 (3H, m), 7.07 (1H, dd), 6.95 (2H, d), 6.34 (1H, dd), 6.14 (1H, dd), 5.59 (1H, d), 2.77 (3H, d), 2.31 (3H, s)
2 4- of embodiment (4- (4- (3- acrylamidos phenylamino)-5- (mesyl) pyrimidine -2 --amino) benzene oxygen Base)-N- picoline -2- formamides (compound 8) preparation
(1) 3- (2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) -5- (mesyl) pyrimidines - 4- bases amino) phenylcarbamate preparation
By 3- (2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) -5- (methyl mercapto) pyrimidines -4- Base amino) phenylcarbamate (0.32g, 0.56mmol) is dissolved in 15mL methanol and 5mL water, then plus sulfuric peroxide Potassium hydrogen phthalate complex salt (oxone) (0.857g, 1.39mmol), system is stirred overnight at normal temperatures, is extracted with 50mL dichloromethane, Twice, organic phase is spin-dried for obtaining 0.34g pale yellow oils 200mL water washings, is directly used in next step.
(2) 4- (4- (4- (3- amino phenyl aminos)-5- (mesyl) pyrimidine -2 --amino) phenoxy group)-N- methyl pyrroles The preparation of pyridine -2- formamides
3- (2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) -5- (methylsulphurs that upper step is obtained Acyl group) pyrimidine-4-yl amino) phenylcarbamate 0.34g is dissolved in the DCM of 10mL, HCl gas reactions are led under ice bath System is spin-dried for by 1h, obtains product crude product 0.36g, is directly used in next step.
(3) 4- (4- (4- (3- acrylamidos phenylamino)-5- (mesyl) pyrimidine -2 --amino) phenoxy group)-N- The preparation of picoline -2- formamides
Upper step product crude product 0.36g is dissolved in the THF of 10mL, DIPEA is added dropwise, until solution ph is 7, at -15 DEG C The THF solution 1.5mL containing acryloyl chloride (0.035g, 0.39mmol) is added dropwise, finishes and reacts 2.5h at a temperature of this.5 drop first are added dropwise System is spin-dried for by alcohol, is prepared liquid phase purifying (methanol: water=57%) and is obtained white solid 13mg.Three step yields：4.1%.
Molecular formula：C27H25N7O5S molecular weight：559.16 mass spectrum (m/z)：559.8(M+H)+.
1H-NMR(d6- DMSO, 400MHz) δ (ppm)：10.19 (2H, s), 8.93 (1H, s), 8.76 (1H, q), 8.50- 8.47 (2H, m), 7.83-7.60 (3H, m), 7.45 (1H, d), 7.38-7.30 (2H, m), 7.23-7.16 (1H, m), 7.12- 6.90 (3H, m), 6.40-6.27 (1H, m), 6.22-6.08 (1H, m), 5.67-5.57 (1H, m), 3.38 (3H, s), 2.77 (3H, d)
3 4- of embodiment (4- (4- (3- acrylamidos phenylamino)-5- (2- methoxyl groups ethylmercapto group) pyrimidine -2 --amino) Phenoxy group)-N- picoline -2- formamides (compound 10) preparation
(1) preparation of S-2- methoxy ethyls thiacetate
By the chloro- 2- Ethyl Methyl Ethers (10.00g, 106mmol) of 1-, potassium carbonate (17.82g, 129mmol), thioacetic acid potassium (14.50g, 127mmol) is added in 50mL DMA, and 5h is stirred at room temperature, and is filtered to remove solid, is poured into 200mL water, acetic acid second Ester extracts, and organic phase is spin-dried for obtaining colorless oil compound, is directly used in next step.
(2) preparation of 2- methoxyl groups ethyl mercaptan
The product that upper step obtains is dissolved in 50mL acetone and 50mL water, sodium hydroxide (5.08g, 12,7mmol) is added, After room temperature reaction stirring 5h, acetone is removed under reduced pressure at 30 DEG C DEG C, obtained aqueous solution is directly used in next step.
(3) preparation of 5- (2- methoxyl groups ethylmercapto group) pyrimidine -2,4 (1H, 3H)-diketone
5-bromouracil (10.00g, 52.4mmol) is added in above-mentioned aqueous solution, back flow reaction 16h.Solution is cold But to -5 DEG C, with concentrated hydrochloric acid tune pH value to 7, solid is precipitated, filters, is dried to obtain faint yellow solid 3.50g, yield 33%.
The preparation of bis- chloro- 5- of (4) 2,4- (2- methoxyl groups ethylmercapto group) pyrimidine
5- (2- methoxyl groups ethylmercapto group) pyrimidine -2,4 (1H, 3H)-diketone (3.00g, 14.8mmol) is added to 50mL POCl3In, the n,N-Dimethylaniline of 2mL is added dropwise, finishes 110 DEG C of reaction 4h.Revolving removes solvent, and 20mL toluene is added again It is spin-dried for system, grease is obtained and pours into ice water, use Na2CO3Aqueous solution is neutralized to neutrality, ethyl acetate extraction, and concentration is organic Phase crosses column (PE/EA=1: 4) and obtains brown solid 620mg, yield 17.5%.
(5) preparation of 3- (the chloro- 5- of 2- (2- methoxyl groups ethylmercapto group) pyrimidine-4-yl amino) phenylcarbamate
By 2,4-, bis- chloro- 5- (2- methoxyl groups ethylmercapto group) pyrimidine (0.60g, 2.51mmol), DIPEA (0.356g, It 2.76mmol) is added in the n-butanol of 10mL, adds 3- aminophenyiaminos t-butyl formate (0.523g, 2.51mmol), System stirs 6h at 100 DEG C.Concentration crosses column (PE: EA=10: 1-1: 2) and obtains white solid 0.50g, yield 48.6%.
(6) 3- (5- (2- methoxyl groups ethylmercapto group) -2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylaminos Base) pyrimidine-4-yl amino) phenylcarbamate preparation
By 3- (the chloro- 5- of 2- (2- methoxyl groups ethylmercapto group) pyrimidine-4-yl amino) phenylcarbamate (0.50g, It 1.22mmol) is dissolved in the tert-pentyl alcohol of 10mL, adds 4- (4- amino-benzene oxygens)-N- picoline -2- formamides (0.296mg, 1.22mmol), acetic acid 2 drip, and 1h is stirred in reaction at 85 DEG C.Concentration, column chromatography (PE: EA=10: 1-2: 1) obtain White solid 0.25g, yield 33.2%.
(7) 4- (4- (4- (3- amino phenyl aminos)-5- (2- methoxyl groups ethylmercapto group) pyrimidine -2 --amino) phenoxy group)-N- The preparation of picoline -2- formamides
By 3- (5- (2- methoxyl groups ethylmercapto group) -2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) Pyrimidine-4-yl amino) phenylcarbamate (0.15g, 0.24mmol) is dissolved in the DCM of 10mL, HCl gas is led under ice bath Precursor reactant 1h, concentration, obtains product crude product 160mg, is directly used in next step.
(8) 4- (4- (4- (3- acrylamidos phenylamino)-5- (2- methoxyl groups ethylmercapto group) pyrimidine -2 --amino) benzene oxygen Base)-N- picoline -2- formamides preparation
The crude product 160mg that upper step obtains is dissolved in the THF of 10mL, DIPEA is added dropwise, until solution ph is 7, -15 DEG C It is lower that the THF solution 0.5mL containing acryloyl chloride (0.022g, 0.24mmol) is added dropwise, it finishes and reacts 15min at a temperature of this.5 drops are added dropwise System is spin-dried for by methanol, is prepared liquid phase purifying (methanol: water=62%) and is obtained white solid 17mg.Two step yields：12.5%.
Molecular formula：C29H29N7O4S molecular weight：571.20 mass spectrum (m/z)：572.2(M+H)+.
1H-NMR(CDCl3, 400MHz) and δ (ppm)：8.40-8.36 (2H, m), 8.25 (1H, s), 8.21 (1H, br s), 8.07 (1H, d), 8.00 (1H, s), 7.69 (1H, d), 7.57 (2H, d), 7.52 (1H, s), 7.38-7.30 (1H, m), 7.29- 7.21 (2H, m), 7.02-6.94 (3H, m), 6.38 (1H, d), 6.18 (1H, dd), 5.63 (1H, d), 3.53 (2H, t), 3.39 (3H, s), 3.03 (3H, d), 2.85 (2H, t)
4 4- of embodiment (4- (4- (3- acrylamidos phenylamino)-5- (2- morpholinyls ethylmercapto group) pyrimidine -2 --amino) Phenoxy group)-N- picoline -2- formamides (compound 11) preparation
(1) preparation of 2- morpholines -4- bases ethylimino thiocarbamate ester
N- (2- chloroethyls) morpholine hydrochloride (9.3g, 0.05mol), thiocarbamide (3.81g, 0.05mol) are dissolved in 95% It in ethyl alcohol (50mL), is heated to reflux 24 hours, cooling, concentration obtains white solid and is directly used in next step.
The preparation of (2) 4,4 '-[dithiobis (ethane -2,1- diyl)] dimorpholines
Crude product (about 0.05mol) obtained by upper step is dissolved in the aqueous solution (30mL) of sodium hydroxide (2g, 0.05mol), It is heated to reflux 2h, cooling, with dilute hydrochloric acid tune pH to 5, ethyl acetate extraction, anhydrous sodium sulfate drying is concentrated to give product 6.02g, Yield 82.4%.
(3) preparation of 2- morpholinyls ethyl mercaptan
4,4 '-[dithiobis (ethane -2,1- diyl)] dimorpholines (6.02g, 20.6mmol) are dissolved in absolute ethyl alcohol In (40mL), sodium borohydride (3.8g, 0.1mol) is added and reacts 12 hours at room temperature, water is added, reduced pressure is gone most of molten Agent, ethyl acetate extraction, anhydrous sodium sulfate drying are spin-dried for, and are obtained oily crude product and are directly used in next step.
(4) preparation of 5- (2- morpholinyls ethylmercapto group) pyrimidine -2,4 (1H, 3H)-diketone
By crude product (about 41.2mol), sodium hydroxide (1.6g, 0.04mol) obtained by upper step and 5-bromouracil (6.36g, It 33.3mmol) is dissolved in heating reflux reaction 5h in water (50mL), it is cooling, add dilute hydrochloric acid to be adjusted to neutrality, white solid is precipitated, takes out Filter, it is dry, obtain product 3.7g, yield 43.2%.
(5) preparation of 4- (2- (2,4- dichloro pyrimidine -5- bases sulfenyl) ethyl) morpholine
5- (2- morpholinyls ethylmercapto group) pyrimidine -2,4 (1H, 3H)-diketone (3.7g, 14.4mol) is dissolved in phosphorus oxychloride In (50mL), back flow reaction 8h is cooling, and after most of solvent is concentrated under reduced pressure, residue is poured into ice water, ethyl acetate extraction It takes, anhydrous sodium sulfate drying, silica gel column chromatography after concentration (petroleum ether: ethyl acetate=5: 1) obtaining faint yellow solid 1.72g, Yield 40.6%.
(6) preparation of 3- (the chloro- 5- of 2- (2- morpholinyls ethylmercapto group) pyrimidine-4-yl amino) phenylcarbamate
By 4- (2- (2,4- dichloro pyrimidine -5- bases sulfenyl) ethyl) morpholine (1.72g, 5.85mmol), 3- aminophenyl ammonia Base t-butyl formate (1.22g, 5.85mmol) and n,N-diisopropylethylamine (908mg, 7.03mmol) are dissolved in tert-pentyl alcohol, are returned 12h is reacted under stream mode, cooling, silica gel column chromatography (petroleum ether: ethyl acetate=3: 1) obtains faint yellow solid after concentration 1.48g, yield 54.4%.
(7) 3- (2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) -5- (2- morpholinyl second sulphur Base) pyrimidine-4-yl amino) phenylcarbamate preparation
By 3- (the chloro- 5- of 2- (2- morpholinyls ethylmercapto group) pyrimidine-4-yl amino) phenylcarbamate (1.48g, 3.18mmol), 4- (4- amino-benzene oxygens)-N- picoline -2- formamides (774mg, 3.18mmol) and two drop glacial acetic acids are molten In n-butanol, 12h is reacted under reflux state, cooling, silica gel column chromatography (petroleum ether: ethyl acetate=1: 1) obtains after concentration White solid 820mg, yield 38.4%.
(8) 4- (4- (4- (3- acrylamidos phenylamino)-5- (2- morpholinyls ethylmercapto group) pyrimidine -2 --amino) benzene oxygen Base)-N- picoline -2- formamides
By 3- (2- (4- (2- (methylcarbamoyl) pyridin-4-yls oxygroup) phenylamino) -5- (2- morpholinyls ethylmercapto group) Pyrimidine-4-yl amino) phenylcarbamate (820g, 1.22mmol) is dissolved in dichloromethane (60mL), slowly it is added dropwise three Fluoroacetic acid (695mg, 6.10mmol) after stirring 6h at room temperature, is dissolved in anhydrous tetrahydro furan (30mL), N is added after reduced pressure, Acryloyl chloride (110mg, 1.22mmol) is slowly added dropwise at -20 DEG C thereto for N- diisopropylethylamine (1.26g, 9.75mmol), Stirring 2 hours is added absolute methanol and is quenched, concentrates, and silica gel column chromatography (petroleum ether: ethyl acetate=2: 1), obtains white solid 177mg, yield 23.1%.
Molecular formula：C32H34N8O4S molecular weight：626.24 mass spectrum (m/z)：627.3(M+H)+
1H-NMR(CDCl3, 400MHz) and δ (ppm)：8.35 (1H, d), 8.04 (1H, q), 7.77 (1H, s), 7.72-7.64 (2H, m), 7.53 (2H, d), 7.15 (1H, s), 7.05 (1H, t), 7.01-6.88 (4H, m), 6.68 (1H, d), 6.41 (1H, Dd), 6.32 (1H, d), 6.24 (1H, dd), 5.72 (1H, d), 4.02 (1H, br s), 3.93-3.87 (2H, m), 3.84 (2H, T), 3.75 (2H, t), 3.65 (2H, t), 3.26 (2H, t), 3.01 (3H, d), 2.95-2.89 (2H, m)
5 N- of embodiment (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methyl mercapto) pyrimidine-4-yls amino) benzene Base) acrylamide (compound 13) preparation
(1) 3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl amino first The preparation of tert-butyl acrylate
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.25g, 0.68mmol) It is dissolved in the tert-pentyl alcohol of 10mL, adds the drop of acetic acid 2,4- (2- methoxy ethoxies) aniline (0.119g, 0.71mmol), reaction 1h is stirred at 85 DEG C.System is spin-dried for, column chromatography (PE: EA=8: 1-2: 1) obtains white solid 0.15g, yield 44.1%.
(2)N4(3- aminophenyls)-N2(4- (2- methoxy ethoxies) phenyl) -5- (methyl mercapto) pyrimidine -2,4- diamines Preparation
By 3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl amino first Tert-butyl acrylate (0.14g, 0.28mmol) is dissolved in the DCM of 10mL, and HCl gas reaction 1h are led under ice bath, system is spin-dried for, is obtained Crude product 0.14g is directly used in next step.
(3) N- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) third The preparation of acrylamide
Upper step product crude product 0.14g is dissolved in the THF of 10mL, DIPEA is added dropwise, until solution ph is 7, at -15 DEG C The THF solution 0.5mL containing acryloyl chloride (0.026g, 0.29mmol) is added dropwise, finishes and reacts 15min at a temperature of this.5 drop first are added dropwise System is spin-dried for by alcohol, is prepared liquid phase purifying (methanol/water=65%) and is obtained white solid 11mg.Two step yields：8.7%.
Molecular formula：C23H25N5O3S molecular weight：451.17 mass spectrum (m/z)：452.2(M+H)+.
1H-NMR(CDCl3, 400MHz) and δ (ppm)：8.24 (1H, s), 8.11 (1H, s), 8.03 (1H, s), 7.55-7.47 (1H, m), 7.45 (2H, d), 7.32-7.27 (1H, m), 7.25-7.20 (1H, m), 7.16-7.04 (2H, m), 6.93 (2H, d), 6.46 (1H, dd), 6.26 (1H, dd), 5.80 (1H, dd), 4.12 (2H, t), 3.76 (2H, t), 3.46 (3H, s), 2.28 (3H, s).
6 N- of embodiment (3- (2- (6- (methoxypyridine -3- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) benzene Base) acrylamide (compound 15) preparation
(1) 3- (2- (6- methoxypyridine -3- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamic acid The preparation of the tert-butyl ester
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.25g, 0.68mmol) It is dissolved in the tert-pentyl alcohol of 5mL, adds the drop of acetic acid 2,6- methoxypyridine -3- amine (0.088g, 0.71mmol) is reacted at 85 DEG C Stir 1h.Reaction is cooled to room temperature, and 10mL ether is added, and filtering obtains pale solid 0.21g, yield 67.6%.
(2)N4(3- aminophenyls)-N2The system of (6- methoxypyridine -3- bases) -5- (methyl mercapto) pyrimidine -2,4- diamines It is standby
By 3- (2- (6- methoxypyridine -3- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamic acid uncle Butyl ester (0.21g, 0.46mmol) is dissolved in the DCM of 10mL, and HCl gas reaction 1h are led under ice bath, system is spin-dried for, product is obtained Crude product 0.22g is directly used in next step.
(3) N- (3- (2- (6- methoxypyridine -3- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) propylene The preparation of amide
Upper step product crude product 0.22g is dissolved in the THF of 10mL, DIPEA is added dropwise, until solution ph is 7, at -15 DEG C The THF solution 0.5mL containing acryloyl chloride (0.042g, 0.46mmol) is added dropwise, finishes and reacts 15min at a temperature of this.5 drop first are added dropwise System is spin-dried for by alcohol, is prepared liquid phase purifying (methanol: water=67%) and is obtained white solid 27mg.Two step yields：14.3%.
Molecular formula：C20H20N6O2S molecular weight：408.14 mass spectrum (m/z)：409.0(M+H)+.
1H-NMR (DMSO-d6,400MHz) δ (ppm)：10.11 (1H, s), 9.25 (1H, s), 8.69 (1H, s), 8.26 (1H, s), 8.18 (1H, s), 7.96 (1H, dd), 7.87 (1H, s), 7.45-7.32 (2H, m), 7.26 (1H, t), 6.58 (1H, D), 6.43 (1H, dd), 6.23 (1H, dd), 5.74 (1H, dd), 3.75 (3H, s), 2.29 (3H, s)
7 N- of embodiment (3- (5- (methyl mercapto) -2- (4- morpholines phenylamino) pyrimidine-4-yl amino) phenyl) acrylamide The preparation of (compound 16)
(1) preparation of 4- (4- nitrobenzophenones) morpholine
P-fluoronitrobenzene (5.00g, 35.4mmol) and morpholine (9.25g, 106mmol) are dissolved in toluene solution, reacted System reflux 5h, is spin-dried for obtaining yellow solid 7.06g, yield 95.8%.
(2) preparation of 4- morpholines aniline
4- (4- nitrobenzophenones) morpholines (2.20g, 10.6mmol) are dissolved in 12mL methanol, nitrogen is replaced, 30mg is added 10% palladium charcoal, three times, reaction stirring 17h, filters off palladium charcoal, is spin-dried for filtrate and obtains pale yellow oil system replacing hydrogen under room temperature 1.65g yield：87.4%.
(3) system of 3- (5- (methyl mercapto) -2- (4- morpholines phenylamino) pyrimidine-4-yl amino) phenylcarbamate It is standby
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.25g, 0.68mmol), It is dissolved in the tert-pentyl alcohol of 5mL, adds the drop of acetic acid 2,4- morpholines aniline (0.121g, 0.68mmol), 1h is stirred in reaction at 85 DEG C. Reaction is cooled to room temperature, and 10mL ether is added, and filtering obtains pale solid 0.27g, yield 77.9%.
(4)N4(3- aminophenyls) -5- (methyl mercapto)-N2The preparation of (4- morpholinyl phenyls) pyrimidine -2,4- diamines
By 3- (5- (methyl mercapto) -2- (4- morpholines phenylamino) pyrimidine-4-yl amino) phenylcarbamate (0.27g, 0.53mmol) is dissolved in the DCM of 10mL, and HCl gas reaction 1h are led under ice bath, system is spin-dried for, crude product is obtained 0.28g is directly used in next step.
(5) preparation of N- (3- (5- (methyl mercapto) -2- (4- morpholines phenylamino) pyrimidine-4-yl amino) phenyl) acrylamide
By N4(3- aminophenyls) -5- (methyl mercapto)-N2(4- morpholinyl phenyls) pyrimidine -2,4- diamines crude product 0.28g is dissolved in In the THF of 10mL, DIPEA is added dropwise, until solution ph is 7, (0.048g, 0.53mmol) containing acryloyl chloride is added dropwise at -15 DEG C FHF solution 0.5mL, finish and react 15min at a temperature of this.5 drop methanol are added dropwise, system is spin-dried for, prepares liquid phase and purifies (methanol : water=66%) obtain white solid 50mg.Two step yields：20.4%.
Molecular formula：C24H26N6O2S molecular weight：462.18 mass spectrum (m/z)：463.3(M+H)+.
1H-NMR (DMSO-d6,400MHz) δ (ppm)：10.14 (1H, s), 9.11 (1H, s), 8.65 (1H, s), 8.16 (1H, s), 7.84 (1H, s), 7.50-7.44 (3H, m), 7.42-7.33 (1H, m), 7.28 (1H, t), 6.72 (2H, d), 6.44 (1H, dd), 6.24 (1H, dd), 5.75 (1H, dd), 3.70 (4H, t), 2.95 (4H, t), 2.28 (3H, s)
8 N- of embodiment (3- (2- (6- (methoxy ethoxy) pyridin-3-yls amino) -5- (methyl mercapto) pyrimidine-4-yl ammonia Base) phenyl) acrylamide (compound 17) preparation
(1) preparation of 2- (2- methoxy ethoxies) -5- nitros
2- chloro-5-nitropyridines (3.00g, 18.9mmol) and 2-methyl cellosolve (2.16g, 28.4mmol) are dissolved in In 10mL DMF, potassium tert-butoxide (2.55g, 22.7mmol) is then added portionwise, system stirs 2h at normal temperatures, then by solution It is added drop-wise in 100mL water, solid, filtering, dry 2.30g brown solids, yield is precipitated：61.4%.
(2) preparation of 6- (2- methoxy ethoxies) pyridine -3- amine
2- (2- methoxy ethoxies) -5- nitros (1.53g, 7.72mmol) are dissolved in 10mL methanol, nitrogen is replaced, adds Enter the palladium charcoal of 35mg 10%, three times, reaction stirring 17h, filters off palladium charcoal, be spin-dried for filtrate and obtain brown system replacing hydrogen under room temperature Grease 1.05g, yield：80.8%.
(3) 3- (2- (6- (2- methoxy ethoxies) pyridin-3-yl amino) -5- (methyl mercapto) pyrimidine-4-yls amino) benzene The preparation of carbamate
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.22g, 0.60mmol) It is dissolved in the tert-pentyl alcohol of 5mL, adds acetic acid 2 and drip, 6- (2- methoxy ethoxies) pyridine -3- amine (0.106g, 0.63mmol), 30min is stirred in reaction at 85 DEG C.Reaction is cooled to room temperature, and system is spin-dried for, and column chromatography (PE: AE=8: 1-2: l) obtains white Solid 0.17g, yield 56.7%.
(4)N4(3- aminophenyls)-N2(6- (2- methoxy ethoxies) pyridin-3-yl) -5- (methyl mercapto) pyrimidine -2, The preparation of 4- diamines
By 3- (2- (6- (2- methoxy ethoxies) pyridin-3-yl amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl T-butyl carbamate (0.17g, 0.34mmol) is dissolved in the DCM of 10mL, and HCl gas reaction 1h are led under ice bath, system is revolved It is dry, product crude product 0.17g is obtained, is directly used in next step.
(5) N- (3- (2- (6- (2- methoxy ethoxies) pyridin-3-yl amino) -5- (methyl mercapto) pyrimidine-4-yls amino) Phenyl) acrylamide preparation
Upper step product crude product 0.17g is dissolved in the THF of 10mL, DIPEA is added dropwise, until solution ph is 7, at -15 DEG C The THF solution 1mL containing acryloyl chloride (0.031g, 0.34mmol) is added dropwise, finishes, reacts 25min at a temperature of this.5 drop first are added dropwise System is spin-dried for by alcohol, is prepared liquid phase purifying (methanol: water=70%) and is obtained white solid 87mg.Two step yields：55.9%.
Molecular formula：C22H24N6O3S molecular weight：452.16 mass spectrum (m/z)：452.9(M+H)+.
1H-NMR (DMSO-d6,400MHz) δ (ppm)：10.12 (1H, s), 9.25 (1H, s), 8.71 (1H, s), 8.25 (1H, s), 8.19 (1H, s), 7.95 (1H, dd), 7.86 (1H, s), 7.44-7.32 (2H, m), 7.26 (1H, t), 6.59 (1H, D), 6.43 (1H, dd), 6.23 (1H, dd), 5.74 (1H, dd), 4.26 (2H, t), 3.60 (2H, t), 3.27 (3H, s), 2.29 (3H, s)
9 N- of embodiment (3- (2- (6- (dimethylamino) pyridin-3-yls amino) -5- (methyl mercapto) pyrimidines -4Base amino) Phenyl) acrylamide (compound 18) preparation
(1) preparation of N, N- dimethyl -5- nitropyridine -2- amine
2- chloro-5-nitropyridines (2.50g, 15.8mmol) and dimethylamine hydrochloride (1.93g, 23.7mmol) are dissolved in In 10mL DMF, potassium tert-butoxide (3.78g, 33.7mmol) is then added portionwise, system stirs 4h at normal temperatures, then by solution It is added drop-wise in 100mL water, solid is precipitated, filter, it is dry, obtain 2.10g light tan solids, yield：79.7%.
(2)N2, N2The preparation of lutidines -2,5- diamines
By N, N- dimethyl -5- nitropyridine -2- amine (1.90g, 11.4mmol) is dissolved in 10mL methanol, replaces nitrogen, Be added 25mg 10% palladium charcoal, system replacing hydrogen three times, under room temperature reaction stirring 17h, filter off palladium charcoal, be spin-dried for filtrate obtain it is brown Color grease 1.25g, yield：79.9%.
(3) 3- (2- (6- (dimethylamino) pyridin-3-yls amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl amino The preparation of t-butyl formate
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.20g, 0.55mmol) It is dissolved in the tert-pentyl alcohol of 5mL, adds the drop of acetic acid 2, N2, N2Lutidines -2,5- diamines (0.079g, 0.58mmol), instead 30min should be stirred at 85 DEG C.Reaction is cooled to room temperature, and 15mL ether is added, and solid is precipitated, and filtering obtains white solid 0.17g, Yield 65.5%.
(4)N4(3- aminophenyls)-N2(6- (dimethylamino) pyridin-3-yl) -5- (methyl mercapto) pyrimidine -2,4- diamines Preparation
By 3- (2- (6- (dimethylamino) pyridin-3-yls amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl amino first Tert-butyl acrylate (0.17g, 0.36mmol) is dissolved in the DCM of 10mL, and HCl gas reaction 1h are led under ice bath, system is spin-dried for, is obtained Product crude product 0.17g is directly used in next step.
(5) N- (3- (2- (6- (dimethylamino) pyridin-3-yls amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) The preparation of acrylamide
Upper step product crude product 0.17g is dissolved in the THF of 10mL, DIPEA is added dropwise, until solution ph is 7, at -15 DEG C The FHF solution 0.5mL containing acryloyl chloride (0.035g, 0.39mmol) are added dropwise, finish, react 15min at a temperature of this.5 drops are added dropwise System is spin-dried for by methanol, is prepared liquid phase purifying (methanol: water=68%) and is obtained white solid 34mg.Two step yields：22.5%.
Molecular formula：C21H23N7OS molecular weight：421.17 mass spectrum (m/z)：422.2(M+H)+.
1H-NMR (DMSO-d6,400MHz) δ (ppm)：10.10 (1H, s), 8.99 (1H, s), 8.60 (1H, s), 8.14 (2H, s), 7.84 (1H, t), 7.79 (1H, dd), 7.40 (2H, d), 7.24 (1H, t), 6.49-6.38 (2H, m), 6.24 (1H, Dd), 5.74 (1H, dd), 2.92 (6H, s), 2.27 (3H, s)
10 N- of embodiment (3- (5- (methylsulfinyl) -2- (4- morpholine phenylaminos）Pyrimidine-4-yl amino) phenyl) third The preparation of acrylamide (compound 19)
By N- (3- (5- (methyl mercapto) -2- (4- morpholines phenylamino) pyrimidine-4-yl amino) phenyl) acrylamide (0.140g, 0.303mmol) is dissolved in 15mL methanol and 5mL water, then plus ammonium persulfate-sodium bisulfate (oxone) (0.186g, 0.303mmol), 25min is reacted at normal temperatures, solution is spin-dried for, and is prepared liquid phase purifying (methanol/water=58%) and is obtained white admittedly Body 45mg.Yield：31%.
Molecular formula：C24H26N6O3S molecular weight：478.18 mass spectrum (m/z)：478.9(M+H)+.
1H-NMR (DMSO-d6,400MHz) δ (ppm)：10.18 (1H, s), 9.54 (2H, s), 8.20 (1H, s), 7.72 (1H, s), 7.55-7.38 (4H, m), 7.29 (1H, t), 6.79 (2H, d), 6.43 (1H, dd), 6.25 (1H, dd), 5.76 (1H, Dd), 3.71 (4H, t), 2.99 (4H, t), 2.95 (3H, s)
11 N- of embodiment (3- (2- (6- methoxypyridine -3- bases amino) -5- (methylsulfinyl) pyrimidine-4-yl ammonia Base) phenyl) acrylamide (compound 20) preparation
By compound N-(3- (2- (6- methoxypyridine -3- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) Acrylamide (0.150g, 0.367mmol) is dissolved in 15mL methanol and 5mL water, then adds ammonium persulfate-sodium bisulfate (oxone) (0.226g, 0.368mmol), reacts 25min at normal temperatures, and solution is spin-dried for, and prepares liquid phase purifying (methanol/water=61%) and obtains To white solid 25mg.Yield：16%.
Molecular formula：C20H20N6O3S molecular weight：424.13 mass spectrum (m/z)：424.9(M)+.
1H-NMR (DMSO-d6,400MHz) δ (ppm)：10.15 (1H, s), 9.68-9.55 (2H, m), 8.33-8.25 (1H, m), 8.22 (1H, s), 7.95 (1H, d), 7.74 (1H, s), 7.41-7.23 (3H, m), 6.73-6.60 (1H, m), 6.42 (1H, dd), 6.23 (1H, dd), 5.75 (1H, dd), 3.78 (3H, s), 2.95 (3H, s)
12 N- of embodiment (3- (2- (6- (dimethylamino) pyridin-3-yls amino) -5- (methylsulfinyl) pyrimidines -4- Base amino) phenyl) acrylamide (compound 21) preparation
By N- (3- (2- (6- (dimethylamino) pyridin-3-yls amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) third Acrylamide (0.150g, 0.356mmol) is dissolved in 15mL methanol and 5mL water, then adds ammonium persulfate-sodium bisulfate (oxone) (0.220g, 0.358mmol) reacts 25min under room temperature, solution is spin-dried for, and prepares liquid phase purifying (methanol/water=51%) and obtains White solid 35mg.Yield：22.5%.
Molecular formula：C21H23N7O2S molecular weight：437.16 mass spectrum (m/z)：437.9(M+H)+.
1H-NMR (DMSO-d6,400MHz) δ (ppm)：10.13 (1H, s), 9.56 (1H, s), 9.50-9.32 (1H, m), 8.17 (2H, s), 7.76 (1H, dd), 7.72-7.66 (1H, dd), 7.43-7.15 (3H, m), 6.60-6.46 (1H, m), 6.41 (1H, dd), 6.24 (1H, dd), 5.75 (1H, dd), 2.96 (6H, s), 2.94 (3H, s)
13 4- of embodiment (4- (4- (3- acrylamidos phenylamino)-5- (methylsulfinyl) pyrimidine -2 --amino) Phenoxy group)-N- picoline -2- formamides (compound 22) preparation
By 4- (4- (4- (3- acrylamidos phenylamino)-5- (methyl mercapto) pyrimidine -2 --amino) phenoxy group)-N- methyl Pyridine-2-carboxamide (0.120g, 0.227mmol) is dissolved in 15mL methanol and 5mL water, and potassium acid sulfate complex salt is then added (oxone) (0.142g, 0.231mmol), reacts 25min at normal temperatures, and solution is spin-dried for, the purifying of preparation liquid phase (methanol/water= 58%) white solid 45mg, yield are obtained：36.5%.
Molecular formula：C27H25N7O4S molecular weight：543.17 mass spectrum (m/z)：543.9(M+H)+.
1H-NMR(d6- DMSO, 400MHz) δ (ppm)：10.18 (1H, s), 9.88 (1H, s), 9.58 (1H, s), 8.77 (1H, q), 8.48 (1H, d), 8.28 (1H, s), 7.82-7.72 (3H, m), 7.45-7.25 (4H, m), 7.15-6.98 (3H, m), 6.35 (1H, dd), 6.17 (1H, d), 5.63 (1H, d), 2.98 (3H, s), 2.77 (3H, d)
14 N- of embodiment (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methylsulfinyl) pyrimidine-4-yls Amino) phenyl) acrylamide (compound 23) preparation
By N- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) third Acrylamide (0.045g, 0.1mmol) is dissolved in open ports reflux 3h in methanol, is spin-dried for, the purifying of preparation solution phase reverse phase (methanol/water= 57%) white solid 9mg is obtained.Yield：19.3%.
Molecular formula：C23H25N5O4S molecular weight：467.16 mass spectrum (m/z)：468.2(M+H)+.
1H-NMR(DMSO-d6, 400MHz) and δ (ppm)：10.17 (1H, s), 9.54 (2H, s), 8.20 (1H, s), 7.73 (1H, s), 7.55-7.45 (3H, m), 7.39 (1H, d), 7.29 (1H, t), 6.82-6.73 (2H, m), 6.42 (1H, dd), 6.24 (1H, dd), 5.75 (1H, dd), 4.00 (2H, t), 3.62 (2H, t), 3.29 (3H, s), 2.95 (3H, s)
15 N- of embodiment (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) -4- (piperazine -1- bases) but-2-enamides (compound 2) preparation
(1) 3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yls amino) phenylamino The preparation of base t-butyl formate
By 3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl amino first Tert-butyl acrylate (5.7g, 11.47mmol) be dissolved in the mixed solution of 50mL dichloromethane and methanol (methylene chloride/methanol=15: 1), hydrosulfuric acid potassium complex salt (7.05g, 11.47mmol) is added portionwise in the solution, TLC monitors raw material and disappears then 100mL dichloromethane and 100mL water, extraction is added, organic phase drying is spin-dried for, crosses silicagel column (methylene chloride/methanol=15: 1) Obtain white solid 5.1g, yield：86.6%.
(2)N4(3- aminophenyls)-N2(4- (2- methoxy ethoxies) phenyl) -5- (methanesulfinyl) pyrimidine -2,4- The preparation of diamines
By 3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yls amino) phenylamino Base t-butyl formate (2.3g, 4.48mmol) is dissolved in 100mL dichloromethane, then be added trifluoracetic acid (2.56g, 22.55mmol) and then 4h is stirred, is spin-dried for, is directly used in next step.
(3) prepared by 4- bromines but-2-ene acyl chlorides
4- bromocrotonic acids (0.718g, 4.35mmol) are dissolved in 20mL dichloromethane, 2 drop DMF are added, then at 0 DEG C When be added 2mL oxalyl chlorides.Acquired solution stirs 2h, is spin-dried for being directly used in next step.
(4) the bromo- N- of 4- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) but-2-enamides preparation
By N4(3- aminophenyls)-N2(4- (2- methoxy ethoxies) phenyl) -5- (methanesulfinyl) pyrimidine -2,4- Two, amine (1.8g, 4.35mmol) are dissolved in the tetrahydrofuran of 20mL dryings, and 4mL DIPEA are added and make pH value to 10, then will 4- bromine but-2-enes acyl chlorides (crude product, about 4.35mmol) is dissolved in 10mL dichloromethane, is slowly added drop-wise in the solution, reaction 2h is stirred at 0 DEG C, is placed spare.
(5) 4- (4- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yls amino) Phenylamino) -4- oxo but-2-enes base) piperazine -1- t-butyl formates preparation
By the bromo- N- of 4- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) but-2-enamides (0.812g, 1.45mmol) and piperazine -1- t-butyl formates (0.323g, 1.74mmol) be dissolved in In the tetrahydrofuran of 10mL dryings, DIPEA (2mL) is added, at 0 DEG C, stirs 0.5h, waits for that raw material disappears, is spin-dried for, cross silica gel Column (methanol=10 DCM/: 1) obtains 0.14g white solids, yield：14.5%.
(6) N- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yls amino) benzene Base) -4- (piperazine -1- bases) but-2-enamides preparation
By 4- (4- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yls amino) Phenylamino) -4- oxo but-2-enes base) piperazine -1- t-butyl formates (0.14g, 0.21mmol), it is dissolved in 10mL dichloromethane, It is slowly added into 0.24g trifluoracetic acids to be spin-dried for after 4h is stirred in reaction, adds the bicarbonate of 100mL dichloromethane and 50mL saturations After being spin-dried for, 2mL methanol is added in sodium solution, extraction, organic phase drying, and recrystallization obtains white solid 0.078g, yield： 65.7%.
Molecular formula：C28H35N7O4S molecular weight：565.2 mass spectrums (m/z)：283.7(M/2+1)+.
1H-NMR(d6- DMSO, 400MHz) δ (ppm)：10.18 (1H, s), 9.58,9.53 (2H, two are unimodal), 8.19 (1H, s), 7.80 (1H, s), 7.65-7.33 (5H, m), 7.26 (1H, t), 6.88-6.65 (3H, m), 6.28 (1H, d), 4.05- 3.95 (2H, m), 3.62 (2H, t), 3.29 (3H, s), 3.05 (2H, d), 2.95 (3H, s), 3.75-3.65 (4H, m), 2.50- 2.22 (4H, m)
16 N- of embodiment (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) -4- morpholinyls but-2-enamides (compound 3) preparation
By the bromo- N- of 4- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) but-2-enamides (0.812g, 1.45mmol), morpholine (0.189g, 2.17mmol) be dissolved in 10mL drying tetrahydrochysene In furans, 2mL DIPEA are added, at 0 DEG C, stir 0.5h, waits for that raw material disappears, is spin-dried for, cross silicagel column (methanol=10 DCM/ : 1) obtain 0.070g white solids, yield：8.5%.
Molecular formula：C28H34N6O5S molecular weight：566.2 mass spectrums (m/z)：284.2(M/2+H)+.
1H-NMR(d6- DMSO, 400MHz) δ (ppm)：10.08 (1H, s), 9.53 (2H, s), 8.20 (1H, s), 7.74 (1H, s), 7.55-7.45 (3H, m), 7.36 (1H, d), 7.27 (1H, t), 6.82-6.65 (3H, m), 6.27 (1H, d), 4.05- 3.95 (2H, m), 3.66-3.54 (6H, m), 3.29 (3H, s), 3.10 (2H, d), 2.95 (3H, s), 2.41-2.34 (4H, m)
Embodiment 17 4- (dimethylamino)-N- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (first sulfenyls Base) pyrimidine-4-yl amino) phenyl) and but-2-enamides (compound 4) preparation
The aqueous solution of the 15mL dimethylamine dichloromethane of 10mL is extracted, the organic phase drying of gained is spare.
By the bromo- N- of 4- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) dichloromethane solution of but-2-enamides (0.812g, 1.45mmol) and 10mL dimethylamine is dissolved in 10mL dryings In tetrahydrofuran, DIPEA (2mL) is added, at 0 DEG C, stirs 0.5h, waits for that raw material disappears, is spin-dried for, cross silicagel column (DCM/ methanol =10: 1) obtaining 0.030g white solids, yield：3.9%.
Molecular formula：C26H32N6O4S molecular weight：524.2 mass spectrums (m/z)：263.1(M/2+H)+.
1H-NMR(d6- DMSO, 400MHz) δ (ppm)：10.19 (1H, s), 9.65-7.50 (2H, m), 8.20 (1H, s), 7.77 (1H, s), 7.60-7.35 (4H, m), 7.28 (1H, t), 6.85-6.68 (3H, m), 6.31 (1H, d), 4.00 (2H, t), 3.62 (2H, t), 3.30-3.15 (5H, m), 2.95 (3H, s), 2.32 (6H, s)
18 N- of embodiment (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) -4- (pyrrolidin-1-yl) but-2-enamides (compound 5) preparation
(1) preparation of 4- (pyrrolidin-1-yl) but-2-ene acid methyl esters
By 4- bromocrotonic acids methyl esters (1.775g, 9.93mmol), it is dissolved in 20mL dichloromethane, then at 0 DEG C into solution Pyrrolidines (1.408g, 19.86mmol) is added, reaction stirring 2h is spin-dried for, is directly used in next step.
(2) preparation of 4- (pyrrolidin-1-yl) but-2-ene acid
4- (pyrrolidin-1-yl) but-2-ene acid methyl esters (1.67g, 9.9mmol) is dissolved in 20mL methanol, is then added The solution of NaOH (0.792g, 19.8mmol), gained stir 2h at 0 DEG C, and ethyl acetate 50mL and water 50mL, extraction is then added It takes, the water phase dilute hydrochloric acid of 2N is adjusted into pH value to 6.0, ethyl acetate 100mL is then added, extracts in three times, gained is organic It is mutually dry, it is spin-dried for, obtains 0.52g pale red grease yield 33.9%.
(3) preparation of 4- (pyrrolidin-1-yl) but-2-ene acyl chlorides
4- (pyrrolidin-1-yl) but-2-ene sour (0.50g, 3.23mmol) is dissolved in 50mL dichloromethane, 2 drops are added DMF, is then slowly added to oxalyl chloride (1.64g, 12.9mmol) at 0 DEG C, and acquired solution stirs 2h, is spin-dried for, is directly used in down One step.
(4) N- (3- (2- (4- (2- methoxy ethoxies) phenylamino) -5- (methanesulfinyl) pyrimidine-4-yls amino) benzene Base) -4- (pyrrolidin-1-yl) but-2-enamides preparation
By N4(3- aminophenyls)-N2(4- (2- methoxy ethoxies) phenyl) -5- (methanesulfinyl) pyrimidine -2,4- Diamines (0.89g, 2.15mmol) is dissolved in the tetrahydrofuran of 10mL dryings, and DIPEA (0.554g, 4.3mmol) is added, and then will 4- (pyrrolidin-1-yl) but-2-ene acyl chlorides (0.56g, 3.23mmol) is dissolved in 10mL dichloromethane, and it is molten to be slowly added drop-wise to this In liquid, 2h is stirred in reaction at 0 DEG C, and LC-MS monitors raw material and disappears, 1mL methanol is then added, is spin-dried for, and crosses silicagel column (DCM/ methanol =10: 1) obtaining 35mg white solids.
Molecular formula：C28H34N6O4S molecular weight：550.2 mass spectrums (m/z)：276.1(M/2+1)+.
1H-NMR(d6- DMSO+HCl, 400MHz) δ (ppm)：11.52 (1H, s), 11.14 (1H, s), 10.93 (1H, s), 10.51 (1H, s), 8.42-8.25 (1H, m), 7.95-7.80 (1H, m), 7.68-7.50 (1H, m), 7.42-7.18 (4H, m), 6.95-6.73 (3H, m), 6.62 (1H, d), 4.12-3.88 (4H, m), 3.68-3.58 (2H, m), 3.50-3.37 (2H, m), 3.28 (3H, s), 3.09-2.93 (5H, m), 2.05-1.80 (4H, m)
19 N- of embodiment (3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) acrylamide (compound 24) preparation
(1) 3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl amino first The preparation of tert-butyl acrylate
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.60g, 1.64mmol), It is dissolved in the tert-pentyl alcohol of 5mL, adds the drop of acetic acid 2,1- methyl-1 H- pyrazoles -4- amine (0.163g, 1.68mmol) is reacted 90 DEG C stirring 13h.It is spin-dried for, crosses silicagel column (PE: EA=2: 1) and obtain 0.52g light red solids, yield：74.4%.
(2)N4(3- aminophenyls)-N2(1- methyl-1 H- pyrazoles -4- bases) -5- (methyl mercapto) pyrimidine -2,4- diamines It prepares
By 3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamic acid The tert-butyl ester (0.52g, 1.22mmol) is dissolved in 20mL dichloromethane, and 2mL trifluoracetic acids are added, and reacts 2h, system is spin-dried for, is obtained To crude product 0.39g, it is directly used in next step.
(3) N- (3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) third The preparation of acrylamide
By N4(3- aminophenyls)-N2(1- methyl-1 H- pyrazoles -4- bases) -5- (methyl mercapto) pyrimidine -2,4- diamines crude product 0.16g is dissolved in the THF of 10mL, and DIPEA (0.126g, 0.98mmol) is added dropwise, until solution ph is 9, is added dropwise at -15 DEG C and is contained third The THF solution 0.5mL of alkene acyl chlorides (0.048g, 0.53mmol) reacts 15min at a temperature of this.5 drop methanol are added dropwise, system is revolved It is dry, it is directly used in next step.
(4) N- (3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methanesulfinyl) pyrimidine-4-yls amino) benzene Base) acrylamide preparation
Previous step crude product is dissolved in 15mL methanol and 5mL water, ammonium persulfate-sodium bisulfate (oxone) is then slowly added into (0.301g, 0.49mmol), reacts 25min at normal temperatures, and solution is spin-dried for, and 30mL dichloromethane and 30mL water is added, and extracts, Organic phase is dried with anhydrous sodium sulfate, is filtered, and filtrate is spin-dried for, and is crossed silicagel column and (it is solid 1) methanol=20: DCM: to be obtained 0.040g whites Body.
Molecular formula：C18H19N7O2S molecular weight：397.1 mass spectrums (m/z)：398.1(M+1)+.
1H-NMR(d6- DMSO+HCl, 400MHz) δ (ppm)：10.28-10.12 (1H, m), 9.68-9.35 (2H, m), 8.20 (1H, s), 7.91-7.15 (6H, m), 6.42 (1H, dd), 6.24 (1H, d), 5.75 (1H, dd), 3.78,3.59 (3H, two It is a unimodal), 2.94 (3H, s)
((2- (1- (2- methoxy ethyls) -1H- pyrazoles -4- bases amino) -5- (methanesulfinyl) is phonetic by 3- by 20 N- of embodiment Pyridine -4- bases amino) phenyl) acrylamide (compound 103) preparation
(1) preparation of 1- (2- methoxy ethyls) -4- nitro -1H- pyrazoles
By 4- nitropyrazoles (2.0g, 17.68mmol), 2- chloroethyl methyl ethers (2.01g, 21.26mmol) and potassium carbonate (4.88g, 35.31mmol) is dissolved in acquired solution reflux 18h in 40mL acetonitriles, is spin-dried for, and crosses silicagel column (PE/EA=4: 1) and obtains 1.5g faint yellow solids, yield：49.5%.
(2) preparation of 1- (2- methoxy ethyls) -1H- pyrazoles -4- amine
1- (2- methoxy ethyls) -4- nitro -1H- pyrazoles (1.2g, 7.01mmol) is dissolved in 25mL methanol, is added The Pd/C of 0.08g 10% three times with hydrogen displacement is stirred overnight under an atmosphere of hydrogen, is filtered after the reaction was complete, methanol washing Filter cake is spin-dried for filtrate and obtains pale red grease 0.80g, yield：80.9%.
(3) 3- (2- (1- (2- methoxy ethyls) -1H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) The preparation of phenylcarbamate：
By 3- (2- chloro- 5- (methyl mercapto) pyrimidine-4-yls amino) phenylcarbamate (0.560g, 1.53mmol), it is dissolved in the tert-pentyl alcohol of 5mL, addition 1- (2- methoxy ethyls) -1H- pyrazoles -4- amine (0.234g, 1.66mmol) with DIPEA (0.292g, 2.26mmol), 18h is stirred at 110 DEG C.It is spin-dried for, crosses silicagel column (PE: EA=2: 1) and obtain 0.42g white solids, yield：58.2%.
(4)N4(3- aminophenyls)-N2(1- (2- methoxy ethyls) -1H- pyrazoles -4- bases) -5- (methyl mercapto) pyrimidine - The preparation of 2,4- diamines trifluoroacetates
By 3- (2- (1- (2- methoxy ethyls) -1H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) benzene Carbamate (0.15g, 0.318mmol) is dissolved in 20mL dichloromethane, and 2mL trifluoracetic acids are added, and reacts 2h, will System is spin-dried for, and obtains crude product 0.12g, is directly used in next step.
(5) N- (3- (2- (1- (2- methoxy ethyls) -1H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yl ammonia Base) phenyl) acrylamide preparation
Upper step crude product 0.118g is dissolved in the THF of 10mL, DIPEA (0.082g, 0.635mmol) is added dropwise, until solution PH value is 9, and the THF solution 0.5mL containing acryloyl chloride (0.029g, 0.320mmol) is added dropwise at -15 DEG C, is finished anti-at a temperature of this Answer 15min.2mL methanol is added dropwise, system is spin-dried for, brownish black grease 0.21g is obtained, is directly used in next step.
(6) N- (3- (2- (1- (2- methoxy ethyls) -1H- pyrazoles -4- bases amino) -5- (methanesulfinyl) pyrimidines -4- Base amino) phenyl) acrylamide preparation
Previous step crude product 0.21g is dissolved in 20mL methanol and 2mL water, the persulfuric acid for being dissolved in 2mL water is then slowly added into Hydrogen potassium complex salt (oxone) (0.301g, 0.49mmol) reacts 25min at normal temperatures, and the sodium thiosulfate of 10mL saturations is added Aqueous solution is quenched, and adds 50mL dichloromethane and 30mL water, and extraction, organic phase is dried with anhydrous sodium sulfate, filters, filtrate rotation It is dry, it crosses silicagel column and (DCM: methanol=20: 1) obtains 0.096g white solids.
Molecular formula：C20H23N7O3S molecular weight：441.2 mass spectrums (m/z)：442.2(M+1)+.
1H-NMR(d6- DMSO+HCl, 400MHz) δ (ppm)：10.25-10.12 (1H, m), 9.68-9.35 (2H, m), 8.20 (1H, s), 7.95-7.17 (6H, m), 6.42 (1H, dd), 6.24 (1H, d), 5.75 (1H, dd), 4.24-3.92 (2H, M), 3.68-3.47 (2H, m), 3.21,3.11 (3H, two unimodal), 2.94 (3H, s)
21 N- of embodiment (3- (2- (1- methyl-IH- pyrazoles -4- bases amino) -5- (methanesulfinyl) pyrimidine-4-yl ammonia Base) phenyl) -4- (pyrrolidin-1-yl) but-2-enamides (compound 104) preparation
(1) the bromo- N- of 4- (3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) benzene Base) but-2-enamides preparation：
4- bromocrotonic acids (0.106g, 0.64mmol) are dissolved in 10mL dichloromethane, 2 drop DMF are added, then at 0 DEG C When be added 2mL oxalyl chlorides, stir 2h, be spin-dried for, it is spare.
By N4(3- aminophenyls)-N2(1- methyl-1 H- pyrazoles -4- bases) -5- (methyl mercapto) pyrimidine -2,4- diamines (0.21g, 0.64mmol) is dissolved in the tetrahydrofuran of 20mL dryings, and DIPEA (0.248g, 1.92mmol) is added so that pH value arrives 10, then 4- bromine crotonyl chlorides are dissolved in 5mL dichloromethane, are slowly added drop-wise in system, 2h is stirred in reaction at 0 DEG C, is put It purchases use.
(2) N- (3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) -4- The preparation of (pyrrolidin-1-yl) but-2-enamides：
0.5mL DIPEA are added in the solution obtained one step up, pyrrolidines (0.055g, 0.77mmol) is stirred at 0 DEG C 0.5h is mixed, 30mL dichloromethane and 30mL water, extraction is added, organic phase is dried with anhydrous sodium sulfate, is filtered, and filtrate is spin-dried for, mistake Silicagel column (DCM: methanol=40: 1) obtains 0.12g white solids, two step yields：40.2%.
(3) N- (3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methanesulfinyl) pyrimidine-4-yls amino) benzene Base) -4- (pyrrolidin-1-yl) but-2-enamides preparation
By N- (3- (2- (1- methyl-1 H- pyrazoles -4- bases amino) -5- (methyl mercapto) pyrimidine-4-yls amino) phenyl) -4- (pyrrolidin-1-yl) but-2-enamides (0.10g, 0.215mmol) are dissolved in 15mL methanol, are then slowly added into and are dissolved in 2mL water Ammonium persulfate-sodium bisulfate (oxone) (0.146g, 0.237mmol), react 25min at normal temperatures, it is full that 10mL be then added It is quenched with hypo solution, then adds 50mL dichloromethane and 30mL water, extracted, organic phase is dry with anhydrous sodium sulfate It is dry, it filters, filtrate is spin-dried for, and crosses silicagel column and (DCM: methanol=20: 1) obtains 0.070g white solids, yield：67.9%.
Molecular weight：480.2 mass spectrums (m/z)：241.2(M/2+1)+, 481.2 (M+1)+.
1H-NMR(d6- DMSO+HCl, 400MHz) δ (ppm)：10.58-10.40 (1H, m), 9.72-9.48 (2H, m), 8.20 (1H, s), 8.02-7.15 (6H, m), 6.85-6.38 (2H, m), 3.86 (2H, s), 3.77 (3H, s), 3.20-3.05 (4H, m), 2.93 (3H, s), 1.90 (4H, m)
22 N- of embodiment (3- (2- (4- (2- methoxy ethoxies) phenyl amino) -5- (methanesulfinyl) pyrimidine-4-yls Amino) phenyl) the amyl- 2- acrylamides (compound 105) of -4- methyl preparation
(1) preparation of the amyl- 2- alkene acyl chlorides of 4- methyl：
It is dissolved in 4- methyl -2- valeric acids (0.50g, 4.38mmol) in 20mL dichloromethane, adds 1 drop DMF, it is cooling To 0 DEG C, oxalyl chloride (1.11g, 8.75mmol) is then slowly added dropwise into the solution, after being added dropwise, stirs 1h at normal temperatures, It is spin-dried for, obtains crude product 0.58g.
(2) N- (3- (2- (4- (2- methoxy ethoxies) phenyl amino) -5- (methyl mercapto) pyrimidine-4-yls amino) benzene Base) the amyl- 2- acrylamides of -4- methyl preparation
N4(3- aminophenyls)-M2(4- (2- methoxy ethoxies) phenyl) -5- (methyl mercapto) pyrimidine -2,4- diamines (0.20g, 0.503mmol) is dissolved in the THF of 15mL, and DIPEA (0.194g, 1.50mmol) is added dropwise, until solution ph is 9 ,- The THF solution 0.5mL containing the amyl- 2- alkene acyl chlorides (0.070g, 0.528mmol) of 4- methyl is added dropwise at 15 DEG C, at a temperature of finishing this React 15min.5 drop methanol are added dropwise, system is spin-dried for, 0.425g brownish black crude oils are obtained, is directly used in next step.
(3) N- (3- (2- (4- (2- methoxy ethoxies) phenyl amino) -5- (methanesulfinyl) pyrimidine-4-yls amino) Phenyl) the amyl- 2- acrylamides of -4- methyl preparation
Previous step crude product 0.425g is dissolved in 10mL methanol and 2mL water, the persulfuric acid for being dissolved in 2mL water is then slowly added into Hydrogen potassium complex salt (oxone) (0.309g, 0.503mmol) reacts 10min at normal temperatures, and the thiosulfuric acid of 15mL saturations is added Sodium water solution is quenched, and adds 50mL dichloromethane and 30mL water, and extraction, organic phase is dried with anhydrous sodium sulfate, filters, filtrate It is spin-dried for, crosses silicagel column and (DCM: methanol=20: 1) obtain 0.060g white solids, two step yields：23.5%.
Molecular formula：C26H31N5O4S molecular weight：509.2 mass spectrums (m/z)：510.1(M+1)+.
1H-NMR(d6- DMSO+HCl, 400MHz) δ (ppm)：10.02 (1H, s), 9.48 (2H, s), 8.19 (1H, s), 7.73 (1H, s), 7.53-7.22 (5H, m), 6.84-6.72 (3H, m), 6.05 (1H, dd), 4.04-3.96 (2H, m), 3.65- The CH of 3.59 (2H, m), 3.29 (3H, s), 2.95 (3H, s), 1.03 (6H, d) allyls are lived by DMSO peak pressures.
With reference to above-mentioned preparation method, it is also prepared for following compound：

Claims (11)

1. general formula (I) compound represented or its pharmaceutically acceptable salt：
Wherein, ring A indicates phenyl；
Ring B indicates phenyl, pyridyl group or pyrazolyl；
L1And L2Expression-NH- independently；
A indicates imino group；
B expressions-CO；
C indicates 1,2- ethenylidenes；
D indicates covalent bond；
E indicates hydrogen atom；
R1Indicate hydrogen atom；
R3Selected from-OC1-4Alkyl ,-O-C1-3Alkylidene-OCH3,-NH-C1-4Alkyl ,-N (C1-3Alkyl)2, C1-4Alkyl or morpholine Base；
R2Expression-L4-R5,
L4Expression-S-C1-3Alkylidene-or-S (O)m,
R5Indicate hydrogen atom, C1-4Alkyl；
M indicates 1 or 2；
The expressions 1 of p and q independently.
2. general formula (I) compound represented or its pharmaceutically acceptable salt：
Wherein, ring A indicates phenyl；
Ring B indicates phenyl, pyridyl group or pyrazolyl；
L1And L2Expression-NH- independently；
A indicates imino group；
B expressions-CO-；
C indicates 1,2- ethenylidenes；
D indicates covalent bond；
E indicates hydrogen atom；
R1Indicate hydrogen atom；
R3Selected from-O (CH3) ,-O (CH2CH3) ,-O-C1-3Alkylidene-OCH3,-NH-CH3,-NH-CH2CH3,-N (C1-3Alkyl) CH3,-N (C1-3Alkyl) CH2CH3, methyl, ethyl or morpholinyl；
R2Expression methylsulfany, ethylsulfanyl, methyl sulphonyl, methylsulfinyl,
The expressions 1 of p and q independently.
3. compound as claimed in claim 2 or its pharmaceutically acceptable salt：
Wherein, ring A indicates phenyl；
Ring B indicates phenyl, pyridyl group or pyrazolyl；
L1And L2Expression-NH- independently；
A indicates imino group；
B expressions-CO-；
C indicates 1,2- ethenylidenes；
D indicates covalent bond；
E indicates hydrogen atom；
R1Indicate hydrogen atom；
R3Selected from-O (CH3) ,-O (CH2CH3) ,-O-CH2CH2-OCH3,-NH-CH3,-NH-CH2CH3, methyl, ethyl or morpholinyl；
R2Indicate unsubstituted ethylsulfanyl, methylsulfany, methyl sulphonyl, methylsulfinyl；
The expressions 1 of p and q independently.
4. compound as claimed in claim 3 or its pharmaceutically acceptable salt：
Wherein, ring A indicates phenyl；
Ring B indicates phenyl or pyridyl group；
L1And L2Expression-NH- independently；
A indicates imino group；
B expressions-CO-；
C indicates 1,2- ethenylidenes；
D indicates covalent bond；
E indicates hydrogen atom；
R1Indicate hydrogen atom；
R3Selected from-O (CH3) ,-O-CH2CH2-OCH3Or morpholinyl；
R2Indicate unsubstituted methylsulfany, methyl sulphonyl, methylsulfinyl；
P indicates 1；
Q indicates 1.
5. compound as described in claim 1 or its pharmaceutically acceptable salt：
Its middle ring A indicates that phenyl, ring B indicate phenyl or pyridyl group；
L1And L2Expression-NH- independently；
A indicates imino group；
B expressions-CO-；
C indicates 1,2- ethenylidenes；
D indicates covalent bond；
E indicates hydrogen atom；
R1Indicate hydrogen atom；
R3Selected from-O (CH3) ,-O (CH2CH3) ,-O-C1-3Alkylidene-OCH3,-NH-CH3,-NH-CH2CH3,-N (C1-3Alkyl) CH3,-N (C1-3Alkyl) CH2CH3, methyl, ethyl or morpholinyl；
R2Expression-L4-R5,
L4Expression-S-C1-3Alkylidene-or-S (O)m,
R5Indicate hydrogen atom or C1-4Alkyl,
M indicates 1 or 2；
P indicates 1,
Q indicates 1.
6. compound or its pharmaceutically acceptable salt, wherein compound are selected from：
7. the preparation method of claim 1-5 any one of them compound or its pharmaceutically acceptable salt, the method packet Include following steps：
Its middle ring A, ring B, L1、L2、R1、R2、R3, any one of a, b, c, d, e, p and q such as claim 1-5 defined.
8. pharmaceutical composition, the composition contains claim 1-6 any one of them compound or its is pharmaceutically acceptable Salt and pharmaceutical acceptable carrier.
9. the pharmaceutical composition of claim 8, wherein the composition also include selected from antitumor agent, immunosuppressor and/or The second therapeutic agent of anti-inflammatory agent.
10. claim 8-9 any one of them pharmaceutical compositions, wherein described pharmaceutical composition are pharmaceutically acceptable One dosage type low temperature.
11. compound as claimed in any one of claims 1 to 6 or its pharmaceutically acceptable salt or such as claim 8-10 Any one of them pharmaceutical composition is being prepared for preventing and/or treating the relevant leukemia of B cell, inflammatory and/or itself exempt from Application in the drug of epidemic disease disease.
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