CN108602834A

CN108602834A - A kind of bruton's tyrosine kinase inhibitor

Info

Publication number: CN108602834A
Application number: CN201780009992.6A
Authority: CN
Inventors: 孟坤; 张建存; 王永钢; 唐勇; 林庆聪; 王骏; 王宗惠
Original assignee: Beijing Shenogen Pharma Group Ltd
Current assignee: Beijing Shenogen Pharma Group Ltd
Priority date: 2016-02-05
Filing date: 2017-01-26
Publication date: 2018-09-28
Anticipated expiration: 2037-01-26
Also published as: CN108602834B; WO2017133630A1; TW201728587A

Abstract

The present invention relates to a kind of bruton's tyrosine kinase inhibitor, which has the structure as shown in following formula (I), and the inhibitor is to having good selectivity inhibition.

Description

A kind of bruton's tyrosine kinase inhibitor

Technical field

The present invention relates to a kind of bruton's tyrosine kinase inhibitor, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxides, belong to field of medicaments.

Background technique

Bruton's tyrosine kinase (Bruton ' s tyrosine kinase, hereinafter referred to as Btk) is a member for belonging to nonreceptor tyrosine kinase Tec family.It is in all hematopoetic cell types other than T lymphocyte, natural killer cells and thick liquid cell, is the important medium of at least three kinds crucial B cell survival mechanisms if having expression in B cell, mast cell and macrophage.Btk is also responsible for the effect of a part to the B cell signal transmission path via B cell antigen receptor (B-cell antigen receptor, i.e. BCR).This multiple action of Btk can make it that B cell malignant tumour be commanded to enter lymphoid tissue, and tumour cell is enable to contact necessary microenvironment and existence.Experimental data shows that Btk also has effect in the signal path of monocyte, macrophage, neutrophil leucocyte and mast cell.The cytokine release that Btk inhibitor can also inhibit monocyte and macrophage to be mediated by Fc also can inhibit the cell degranulation mediated by FcR.The shortage of Btk, which has shown that, can block B cell antigen receptor signal transduction, therefore the compound of the inhibitory activity with Btk can be used as the related disease for blocking B cell and/or mast cell mediated, such as: effective treatment method " the immune summary (International Reviews of Immunology) in the world " of cancer, autoimmune disease, thrombotic disease and inflammatory disease etc., 2012,31,119-132；" arthritis research and treatment (Arthritis Research& Therapy) ", 2011,13, R115；" clinical immunization (Clin.Exp.Immunol.) " 1993,94,459；Chem. " pharmaceutical chemistry (MedChem.) " 2007,2,58-61.It is that WO2008121742 is disclosed with the Btk inhibitor such as following formula (i) in international publication number:

When the L in formula (i) structure is selected from O, Ar is selected from benzene, and Y is selected from piperidines, and Z is selected from carbonyl, Rⁱ、RⁱⁱAnd RⁱⁱⁱWhen being selected from H, existing compound is obtained according to Shandong for Buddhist nun (Ibrutinib；Trade name: Imbruvica), there is the structure as shown in following formula (ii):

Replacing Buddhist nun according to Shandong is on November 13rd, 2013, and the treatment of U.S. FDA approval is for treating lymphoma mantle cell.In addition to this, very big potentiality are also shown in terms for the treatment of chronic lymphocytic leukemia and Huppert's disease for Buddhist nun according to Shandong.

Another Btk inhibitor is disclosed in Publication No. CN102918040, there is the structure as shown in following formula (iii):

Compound with the structure is other than with Btk selective inhibitory activity, or the compound that metabolic stability is excellent, can avoid hepatotoxicity etc., therefore can be used as that safety is excellent, therapeutic agent with the B cells such as non-Hodgkin lymphoma and/or mast cell associated disease.

The Btk inhibitor with following formula (iv) is disclosed in the patent application of Publication No. WO2015048689:

The compound is also disclosed in text for treating the purposes of the diseases such as active immunity, abnormal immune and cancer.

With the continuous deepening of research, the structure of Btk inhibitor and its indication range also have deeper variation.

Summary of the invention

The purpose of the present invention is to provide a kind of bruton's tyrosine kinase inhibitor, the selective inhibitory effect of the inhibitor, for non-Hodgkin lymphoma, wherein it is preferred that B cell non-Hodgkin lymphoma, Such as: diffusivity large B cell lymphoid tumor, human B lymphocytes' tumor, lymphoma mantle cell, small lymphocyte lymthoma, Walden Si Telun (family name) macroglobulinemia (WM) and B cell chronic lymphocytic leukemia have good inhibitory effect.

One aspect of the present invention provides a kind of with logical formula (I) compound represented, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide:

Wherein: W is nitrogenous saturated heterocyclyl, the benzene methylene (C of 4-6 member₃-C₆) naphthenic base or [3.3-5] miscellaneous loop coil base containing nitrification；

R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；Substituted-phenyl replaces the substituent group on nitrogen-containing hetero phenyl that can be each independently selected from halogen, (C₁-C₄) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄) one of halogenated alkoxy or a variety of；

Any one integer between n=0-4；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optics pair Isomers, the mixture of enantiomter, racemic modification or N- oxide are reflected, as shown in formula (II):

Wherein, W is nitrogenous saturated heterocyclyl, the benzene methylene (C of 4-6 member₃-C₆) naphthenic base or [3.3-5] miscellaneous loop coil base containing nitrification；

R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (III):

Wherein, R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (IV):

Wherein, the R²Selected from phenyl, substituted-phenyl, benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkoxy, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkane Oxygroup andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (V):

Wherein, X, Y are each independently selected from CH or N, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (VI):

Wherein, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (VII):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (VIII):

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (IX):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X):

Wherein, X, Y and Z are each independently selected from CH or N；R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄One or more of) halogenated alkoxy.

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (XI):

Wherein, R¹Selected from phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkoxy, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " be selected from phenyl, substituted-phenyl, One or more of nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (XII):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X III):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X IV):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X V):

Wherein, X, Y and Z are each independently selected from CH or N；R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X VI):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X VII):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X VIII):

Wherein, R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X IX):

Wherein, R²Selected from phenyl, substituted-phenyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

R¹Selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl or (C₁-C₄) halogenated alkoxy；

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X X):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X XI):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X XII)

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X X III):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (X X IV):

Preferably, the compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, wherein the compound is selected from one or more of following compound:

1- (3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- tolylthiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- methyl -4- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- (Phenoxymethyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((2- methyl) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((m- methyl) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((p- methyl) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((2- methoxyl group) phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((3- methoxyl group) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((4- methoxyl group) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- ((2- cyano) Phenoxymethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- ((4- cyano) Phenoxymethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- ((3- cyano) Phenoxymethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- (2- methoxybenzene) oxygen methylthiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- (3- methoxybenzene) oxygen methylthiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- benzene thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (Phenoxymethyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((pyrimidine-4-yl oxygroup) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d]-pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- ((2- methoxyl group -4- methylphenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- ((2- methoxyl group -4- chlorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((2- methoxyl group -4- cyano-benzene oxygen) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((2- chloro-5-methoxyl phenoxy group) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((3- chloro-5-methoxyl phenoxy group) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((3- methoxyl group -4- cyano-benzene oxygen) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (2- methylenedioxy phenoxy methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- toloxyl methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- aminomethyl phenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3,4- 3,5-dimethylphenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- methoxyphenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- methoxyphenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- tolylthiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- tolylthiophene -3- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- trifluoromethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- benzotrifluoride) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3,5- dimethylbenzene) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- tolylthiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (2- fluorophenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- fluorophenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- fluorophenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (3- aminomethyl phenyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (4- methoxyphenyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- methoxyphenyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((2- fluorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((3- fluorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((4- fluorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (phenoxymethyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base)-piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (phenethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base)-piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) trans- 1- (3- (4- amino -3- (2- styrene thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- ((3- methylbenzene) oxygen methyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (anilino- formoxyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (2- pyridine amido formacyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (3- pyridine amido formacyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (4- pyridine amido formacyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (anilino- formoxyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (2- pyridine amido formacyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (3- pyridine amido formacyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (4- pyridine amido formacyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (anilino- formoxyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (pyridine -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (pyridin-3-yl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (pyridine -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (pyridin-3-yl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (6- cyanopyridine -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base)-picolinamide；

(R) -6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base)-Niacinamide；

(R)-N- (6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base) pyridine -2- base) acetamide；

(R)-N- (6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base) pyridin-3-yl) acetamide；

(R) -1- (3- (4- amino -3- (5- (6- picoline -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (5- picoline -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone.

The present invention also provides a kind of such as following formula (X X V) compound represented or the salt of the compound,

Wherein, R¹Or R²Selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene Asia First (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；R⁷Selected from H orWherein R³Selected from one of trifluoromethyl, tert-butoxy and benzyloxy；Work as R⁷When for H, the salt of formula (X X V) compound can be one of hydrochloride, sulfate, acetate and trifluoroacetate.Formula (X X V) compound or its salt is the midbody compound of formula (I)-formula of the present invention (X X IV) compound.

Preferably, wherein the compound is as follows:

Wherein, R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；R⁷Selected from H orWherein R³Selected from trifluoromethyl, one of tert-butoxy and benzyloxy；Work as R⁷When for H, the salt of formula (X X VI) compound can be one of hydrochloride, sulfate, acetate and trifluoroacetate.

Preferably, wherein the compound is as follows:

Wherein, R¹Selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；R⁷Selected from H orWherein R³Selected from trifluoromethyl, one of tert-butoxy and benzyloxy；Work as R⁷When for H, the salt of formula (X X VII) compound can be one of hydrochloride, sulfate, acetate and trifluoroacetate.

The present invention also provides a kind of such as following formula (X X VIII) compounds represented, wherein the compound is as follows:

Wherein, R⁸Selected from H, Br or boronate, R⁹Selected from H, methyl, cyano, amine formyl or acetamido.Formula (X X VIII) compound is the midbody compound of formula (I)-formula (X X IV) compound.

Preferably, wherein shown in the compound such as following formula (X X IX):

Wherein, R⁸Selected from H, Br or boronate, R⁹Selected from H, methyl, cyano, amine formyl or acetamido.

Preferably, wherein shown in the compound such as following formula (X X X):

Preferably, wherein shown in the compound such as following formula (X X XI):

Preferably, wherein shown in the compound such as following formula (X X XII):

The present invention also provides compound of the present invention, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide in preparation for treating and including the purposes in malignant tumour, autoimmune disease and anaphylactia related drugs.

Preferably, wherein the malignant tumour includes one or more of lymthoma, plasmacytoma and leukaemia.

Preferably, wherein the lymthoma includes non-Hodgkin lymphoma, follicular lymphoma, covers carefully One or more of born of the same parents' lymthoma, small lymphocyte lymthoma, mantle cell lymphoma, intravascular maxicell type B cell lymphoma, Burkitt lymphoma, AIDS related lymphoma and marginal zone B-cell lymphoma.

Preferably, the non-Hodgkin lymphoma includes B cell non-Hodgkin lymphoma.

It is highly preferred that the B cell non-Hodgkin lymphoma includes one or more of diffusivity large B cell lymphoid tumor and human B lymphocytes' tumor.

Preferably, wherein the autoimmune disease includes one or more of arthritis, rheumatism, inflammatory enteritis and lupus erythematosus.

Further aspect of the present invention provides a kind of bruton's tyrosine kinase inhibitor combination, including the compound of the present invention, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide.

Figure of description

The disease of growth Fig. 1 shows compound through the invention to rheumatic arthritis mouse action time, mouse improves situation.

Specific embodiment

Following embodiment is only used for illustrating the present invention, is not used in the limitation present invention, modification, change, modification for being made in the scope of the present invention etc. are all within the scope of the present invention.

Unless otherwise stated, there is provided in the terms " bruton's tyrosine kinase inhibitor " includes having formula (I), formula (II), formula (III), formula (IV), formula (V), formula (VI), formula (VII), formula (VIII), formula (IX), formula (X), formula (XI), formula (XII), formula (XIII), formula (XIV), formula (XV), formula (X VI), formula (XVII), formula (XVIII), formula (X IX), formula (X X), formula (X X I), formula (X X II), formula (XXIII), the compound of formula (X X IV) structural formula, every kind of compound includes the different stereoisomers with identical structural formula, stereoisomer therein further includes that optical isomer and geometry are different Structure body, optical isomer are also referred to as enantiomter, and geometric isomer is also referred to as cis-trans-isomer.

Optical enantiomorphs are the optical enantiomorphs with chiral centre mirror image each other.

The mixture of optical enantiomorphs refers to that two kinds of chiral each other optical enantiomorphs are mixed with different molar ratios, obtained mixture.

Racemic modification refers to that two kinds of chiral each other optical enantiomorphs are mixed with identical molar ratio, Because intermolecular interaction causes optical activity to be offset, obtained mixture is known as racemic modification.

Cis-trans-isomer refers to: identical atom is located at that carbon-carbon double bond is ipsilateral and two kinds of isomers of heteropleural, and identical atom is located at that carbon-carbon double bond is ipsilateral referred to as cis-, and it is referred to as trans- that same atoms are located at carbon-carbon double bond heteropleural.

Unless otherwise stated, term herein " heterocycle " refers to that one or more carbon atoms for constituting ring of naphthenic base substitute to form heterocycle by the hetero atom other than carbon, and the hetero atom includes but not limited to nitrogen-atoms, oxygen atom and sulphur atom etc.." naphthenic base " includes but not limited to phenyl, hexamethylene etc.." nitrogen heterocycle " refers to that the carbon atom for constituting ring substitutes by one or multiple nitrogen-atoms and to form heterocycle, when the heterocycle of formation is saturated heterocyclyl, referred to as " nitrogenous saturated heterocyclyl "；When the heterocycle of formation is unsaturated heterocycle base, referred to as " nitrogenous unsaturated heterocycle base ".

Unless otherwise stated, term herein " benzene methylene (C₃-C₆) naphthenic base " refer to a hydrogen atom of methyl on benzyl by (C₃-C₆) naphthenic base substitution, the structure of benzene methylene naphthenic base is formed, wherein two groups being connected with naphthenic base include but not limited to adjacent, alternate and opposite position.

Unless otherwise stated, term herein " the miscellaneous loop coil base containing nitrification " refers to that two saturated cyclic alkyls share a carbon atom, forms saturation loop coil.The carbon atom (carbon atom of non-common) on loop coil is saturated by nitrogen-atoms, and it is saturated loop coil group adjacent thereto to constitute containing the miscellaneous loop coil base of nitrification, it is wherein saturated the atom number being related to when the name of miscellaneous loop coil to be determined according to the number of the carbon atom or nitrogen-atoms that form each ring skeleton, without the public carbon atom of two rings.

Unless otherwise stated, term herein " benzene (C₁-C₄) alkyl " refer to a hydrogen atom on phenyl ring by (C₁-C₄) alkyl substitution, the benzene (C of formation₁-C₄) alkyl structure, it include but not limited to benzyl, phenethyl, phenylpropyl, benzene isopropyl and benzene butyl etc..

Unless otherwise stated, term herein " benzene (C₂-C₄) alkynyl " refer to a hydrogen atom on phenyl ring by (C₂-C₄) alkynyl substituted, the benzene (C of formation₂-C₄) alkynyl structure, it include but not limited to phenylacetylene, phenyl-allylene, benzene butine etc..Unless otherwise stated, term herein " substituted-phenyl " is referred to by the hydrogen atom on other atoms or group substituted-phenyl other than hydrogen atom, the obtained phenyl with substituent group.

Unless otherwise stated, term " benzene oxyalkyl " herein refers to that phenyl is connected with alkyl by oxygen, the group of formation, and the group is connected by alkyl with extraneous other groups, includes but not limited to: Phenoxymethyl, benzene oxygen ethyl, benzene oxygen propyl group etc..

Unless otherwise stated, term herein " substituted benzene oxygen alkyl " refer to the phenyl of benzene oxyalkyl by other than hydrogen atom other atoms or group replace, form the benzene oxyalkyl for having substituent group.

Unless otherwise stated, the " (C that amido replaces herein₂-C₄) alkenyl " refer to (C₂-C₄) alkenyl one or more hydrogen atoms replaced by one or more amidos after the obtained (C with amido₂-C₄) alkenyl, " hydrogen atom " therein both include alkenyl on hydrogen atom and also including non-alkenyl such as: the hydrogen atom on methyl, methylene group, " amido " are the organic amino groups formed after the hydrogen atom of ammonia is substituted by alkyl.

Unless otherwise stated, " benzene (C₂-C₄) alkenyl " refer to a hydrogen atom on phenyl ring by (C₂-C₄) alkenyl replace, obtain (the C with phenyl ring₂-C₄) alkenyl, including and be not limited to: styryl, cinnamyl group, benzene isopropenyl, benzene cyclobutenyl etc..

Unless otherwise stated, term " nitrogen-containing hetero phenyl " herein refers to one on phenyl or more than one carbon atom is replaced by nitrogen-atoms, the phenyl for having nitrogen heteroatom is formed, includes but not limited to: pyridyl group, metadiazine base, pyrazine base etc..

Unless otherwise stated, term herein " plasmacytoma " is one group of tumor disease caused by Monoclonal plasmacytosis, including Huppert's disease, primary macroglobulinaemia.

In certain embodiments, study subject of the invention can be mammal, such as dog, cat, ox, sheep, horse or people, preferably people.The required therapeutic dose of drug of the invention is according to the variation of disease specific and can be readily determined by those of ordinary skill in the art.

In some embodiments, one or more compounds of the invention can be used in combination to each other, also it can choose and the compound of the present invention be used in combination with any other active agent, it is used to prepare bruton's tyrosine kinase inhibitor, if using one group of compound, these compounds simultaneously, respectively or in an orderly manner can be administered study subject.

In some embodiments, the compound of the present invention can be used with one or more other anti-cancer agent in conjunction.Can associated with anticancer agent include but is not limited to according to Shandong replace Buddhist nun, lestaurtinib, Tarceva, Lapatinib, linatinib, Lapatinib, Si Dinibu, Axitinib, pazopanib, Sorafenib, VEGF Trap, Rituximab, alemtuzumab, bevacizumab, Victibix, Herceptin, alkylating agent, nitrogen shellfish drug, antifol, purine antagonist, Pyrimidine antagonists, spindle poison, topoisomerase enzyme inhibitor, inducer of apoptosis, angiogenesis inhibitors, podophyllotoxin, nitroso ureas, antimetabolite, protein synthesis inhibitor, kinase inhibitor, antiestrogen, cis-platinum, carboplatin, interferon, asparaginase, Leuprorelin, Flutamide, megestrol acetate, mitomycin, bleomycin, adriamycin, Irinotecan and taxol.In some embodiment, the anticancer agent is antiestrogen, such as tamoxifen and fulvestrant (ICI182,780).

In some embodiments, pharmaceutical composition of the invention can also be used for treatment Animal diseases.Common animal doctor can according to working experience by a kind of compound of the invention or its can be for animals salt its can be for animals solvent or prodrug be administered with suitable acceptable dosage form.Animal doctor can determine to the most suitable administration route of a certain animal.

The compound of the present invention is prepared by following route 1:

In above synthetic route, 5- amido -1H- pyrazoles -4- cyano (compound 1 ') and amitraz hydrochloride as starting material is at 70-90 DEG C, it is preferred that reaction obtains 1H- pyrazoles [3 under the conditions of 80 DEG C, 4-d] pyrimidine -4- amine (compound 2 '), organic solvent (such as, dimethylformamide, acetic acid) in reacted at 80 DEG C with N- N-iodosuccinimide or lodine chloride, thus it carries out, iodide reaction occurs and obtains iodo- 1H- pyrazoles [3, the 4-d] pyrimidine -4- amine (compound 3 ') of 3-.

Reaction in reaction route 1 in step 3 is well known, the iodo- 1H- pyrazoles [3 of 3-, 4-d] pyrimidine -4- amine (compound 3 ') is in organic solvent, as in tetrahydrofuran (THF) with hydroxy compounds 4 ', in the presence of triphenylphosphine and diisopropyl azodiformate, it is reacted at 50-70 DEG C, bimolecular nucleophilic substitution (Mitsunobu substitution) reaction thus occurs, obtains compound 5 '.Compound 4 ' is the raw material with Chirality, it therefore, can be according to the polarimetry nature of target compound during synthesis, select the compound 4 ' of appropriate optical activity as raw material, the reaction intermediate compound obtained from is also provided with the identical polarimetry nature with compound 4 '.Compound 5 ' and compound 6 ', in organic solvent (dimethyl methyl Amide, glycol dimethyl ether, tetrahydrofuran and Isosorbide-5-Nitrae-dioxane (Isosorbide-5-Nitrae-dioxane)) in, in Metal Palladium, such as: including tetrakis triphenylphosphine palladium Pd (PPh₃)₄, double phenyl phosphorus dichloro palladium Pd (Pd (PPh₃)₂Cl₂[1,1 '-bis- (diphenylphosphine) ferrocene] palladium chloride Pd (dppf) Cl₂In under any one or a few catalysis, reacted at 60-80 DEG C, thus carry out coupling reaction occurs to obtain compound 7 '.

The reaction of step 5 is well known, compound 7 ' in reaction route 1, in organic solvent (methylene chloride, tetrahydrofuran); in the presence of trifluoroacetic acid or hydrochloric acid; it at 0 DEG C to reacting at room temperature, thus carries out, carries out deprotection reaction and obtain compound 8 '.

The reaction of step 5 in reaction route 1 is well known, compound 8 ', in organic solvent (methylene chloride or dimethylformamide), with corresponding carboxylic acid, or acyl chlorides at 0 DEG C to reacting at room temperature, thus carry out, occur condensation reaction obtain compound 9 '.

Starting material 5- amino -1H- pyrazoles -4- cyano CAS is purchased from Shanghai Hai Qu Chemical Co., Ltd. for 16617-46-2.

It is 85275-45-2 that compound 4 ', which is 1- tertbutyloxycarbonyl -3- hydroxy piperidine CAS, and (S) -1- tertbutyloxycarbonyl -3- hydroxy piperidine CAS is 143900-44-1 or (R) -1- tertbutyloxycarbonyl -3- hydroxy piperidine CAS is 143900-43-0 purchased from Shanghai Shao Yuan reagent Co., Ltd.

Compound 6 ' is made in accordance with the following methods: the tetrahydrofuran solution of corresponding bromo substituted thiophene compound is added drop-wise in a nitrogen environment in tetrahydrofuran (THF) solution of subzero 70 DEG C of n-BuLi (nBuLi).It is reacted at subzero 70 DEG C 2 hours, methyl borate (B (OMe) is then added₃), the reaction was continued at subzero 70 DEG C 30 minutes, is warmed to room temperature in the hydrochloric acid for importing 1mol/L and stirs 10 minutes.The organic phase being extracted with ethyl acetate is dry, is concentrated under reduced pressure to give target boronic acid compounds crude product, because its is unstable, is directly used in without purification and reacts in next step.

In reaction in the present specification, the reaction of heat tracing is such as common in the art that water-bath or oil bath can be used to carry out.In reaction in the present specification, reaction product can use common means of purification, such as the fraction that normal pressure or vacuum distillation are obtained, and using the high performance liquid chromatography of silica gel, thin-layer chromatography, the method washed etc. is purified.Purifying can carry out in each reaction, can also be operated after multistep reaction.

Embodiment 1

The preparation of 1H- pyrazoles [3,4-d] pyrimidine -4- amine (compound 2 ')

At room temperature to 5- amino -1H- pyrazoles -4- cyano (compound 1 ') (20g, 185.2mmol) Amitraz hydrochloride (16.4g is added in ethyl alcohol (500mL) solution, 203.7mmol), then it is reacted 10 hours at 80 DEG C, it is cooled to room temperature, by the way that solvent is concentrated under reduced pressure, vacuum drying obtains target product (21.3g, 85% yield) after obtained solid product is washed with water, that is 1H- pyrazoles [3,4-d] pyrimidine -4- amine (compound 2 ').

Thin-layer chromatography (TLC) detection: R_f0.5 (methylene chloride: methanol=5: 1, phase is unfolded)

Mass Spectrometer Method: MS (ESI) m/z 136 (M+1)；

Nuclear magnetic resonance: 1HNMR (400MHz, DMSO-d6), 6.96 (br s, 2H), 7.68 (s, 1H), 8.35 (s, 1H), 13.24 (s, 1H).

The preparation of iodo- 1H- pyrazoles [3, the 4-d] pyrimidine -4- amine (compound 3 ') of embodiment 2:3-

At room temperature to 1H- pyrazoles [3,4-d] pyrimidine -4- amine (20g, lodine chloride ICl (24g is added in acetic acid (500mL) solution 148mmol), 148mmol), then it is reacted 10 hours at 80 DEG C, it is cooled to room temperature, by the way that solvent is concentrated under reduced pressure, obtained solid product successively uses Na₂CO₃Saturated solution, Na₂SO₃Vacuum drying obtains iodo- 1H- pyrazoles [3, the 4-d] pyrimidine -4- amine of 3- (compound 3 ', 30.9g, 80% yield) after saturated solution and water washing.Thin-layer chromatography detection: TLC:R_f0.5 (methylene chloride: methanol=5: 1)

Mass Spectrometer Method: MS (ESI) m/z 262 (M+1)；

Magnetic resonance detection: 1HNMR (400MHz, DMSO-d6), 6.98 (br s, 2H), 8.35 (s, 1H), 13.20 (s, 1H).

The preparation of embodiment 3 (compound 4 ')

When ring 2 is piperidines, m=0, compound 4 ' are as follows: 1- tertbutyloxycarbonyl -3- hydroxy piperidine

Compound 5 ' are as follows: tert-butyl -3- (iodo- 1H- pyrazoles [3, the 4-d] pyrimidine -1- base of 4- amino -3-) piperidines -1- formic acid esters

At room temperature to the iodo- 1H- pyrazoles [3 of 3-, 4-d] pyrimidine -4- amine (compound 3 ') (10g, 1- tertbutyloxycarbonyl -3- hydroxy piperidine (compound 4 ') is sequentially added in tetrahydrofuran (150mL) solution 38.3mmol), (9.25g, 46mmol) (according to specific needs, select be suitble to optical activity 1- tertbutyloxycarbonyl -3- hydroxy piperidine), diisopropyl azodiformate (9.30g, 46mmol) and triphenylphosphine (12.06mmol, 46mmol).Then it is reacted 5 hours at 60 DEG C, it is cooled to room temperature, by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure, eluting solvent is methylene chloride: methanol (40: 1- > 10: 1) obtains target product tert-butyl -3- (iodo- 1H- pyrazoles [3, the 4-d] pyrimidine -1- base of 4- amino -3-) piperidines -1- formic acid Ester (compound 5 ', 2.8g, yield: 75%).

Thin-layer chromatography detection: TLC:R_f0.5 (methylene chloride: methanol=10: 1) Mass Spectrometer Method: MS (ESI) m/z 445 (M+1)；Magnetic resonance detection: 1HNMR (400MHz, CDCl₃) 1.67 (s, 9H)；1.70-2.40 (m, 4H), 2.95-4.95 (m, 5H), 6.95 (br s, 2H), 8.31 (s, 1H).Embodiment 4

In product 9 ' in reaction route 1, work as R⁵、R⁶It is H, ring 1 is benzene, L₁For methoxyl group, wherein methyl moiety is connected with thiophene, and oxygroup is connected with benzene, L₁It is connected with 5 of thiophene, 2 of thiophene are connected with 3 of pyrazole ring, m=0, and ring 2 is piperidine ring, and 3 of piperidine ring are connected with 1 nitrogen of pyrazole ring, and 1 nitrogen of piperidine ring is connected with carbonyl, R³For vinyl, prepare as follows

Reaction route 2:

Wherein Et₃N indicates that triethylamine, EDCl indicate 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride.

Work as R⁵、R⁶It is H, ring 1 is benzene, L₁For methoxyl group, wherein methyl moiety is connected with thiophene, and oxygroup is connected with benzene, L₁It is connected with 5 of thiophene, 2 of boron and thiophene are connected, compound 6 ' are as follows: (5- (Phenoxymethyl) thiophene -2- base) boric acid.

(5- (Phenoxymethyl) thiophene -2- base) boric acid of compound 6 ' is to compound 1:1- (3- (4- amino - 3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- The preparation of alkene -1- ketone

I. (5- (Phenoxymethyl) thiophene -2- base) boric acid preparation method

At room temperature to 2- bromothiophene -5- methanol (2g, 10mmol) phenol (1.46g is sequentially added in tetrahydrofuran (50mL) solution of (CAS is that 79387-71-6 is purchased from the smooth Science and Technology Co., Ltd. of Haitai), 15mmol), diisopropyl azodiformate (3.1g, 15mmol) and triphenylphosphine (4g, 15mmol).Then it reacts 10 hours at room temperature, by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure, eluting solvent is ethyl acetate: petroleum ether (1: 20- > 1: 10) obtains 2- bromo- 5- (Phenoxymethyl) thiophene 1.95g, yield: 70%).

Thin-layer chromatography detection: TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 15)

Mass Spectrometer Method: MS (ESI) m/z 269 (M+1)；

Magnetic resonance detection: 1HNMR (400MHz, CDCl₃) 5.20 (s, 2H), 6.50-6.75 (m, 2H), 7.00-7.45 (m, 5H).

2- bromo- 5- (Phenoxymethyl) thiophene (1.90g, tetrahydrofuran (10mL) 7mmol) is dissolved in be added drop-wise in a nitrogen environment in tetrahydrofuran (15mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 5.6mL, 2.5mol/L).It is reacted at -70 DEG C 2 hours, methyl borate (3.67g, 35mmol) then is added, the reaction was continued at -70 DEG C 30 minutes, is warmed to room temperature stirring 10 minutes in the hydrochloric acid (30mL) for importing 1mol/L.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, by being concentrated under reduced pressure to give (5- (Phenoxymethyl) thiophene -2- base) crude boronic acid 1.25g, since product easily decomposes, it is directly used in without purification and reacts in next step.

Thin-layer chromatography detection: TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 5) Mass Spectrometer Method: MS (ESI) m/z 235 (M+1)；

II. tert-butyl -3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] Pyrimidine -1- base) piperidines -1- formic acid esters preparation method

At room temperature to tert-butyl -3- (the iodo- 1H- pyrazoles [3 of 4- amino -3-, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (200mg, (5- (Phenoxymethyl) thiophene -2- base) boric acid (211mg is sequentially added in the solution of glycol dimethyl ether (5mL), water (2mL) 0.45mmol), 0.90mmol), four (triphenyl phosphorus) palladium (26mg, 0.02mmol), Na₂CO₃(143mg, 1.35mmol).Then it is reacted 16 hours at 80 DEG C, is cooled to room temperature, pours into 50mL water, be extracted with ethyl acetate, extract organic liquor anhydrous Na₂SO₄After drying, by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure, eluting solvent is petroleum ether: ethyl acetate (8: 1 → 2: 1) obtains target product tert-butyl 3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- Base) piperidines -1- formic acid esters (160mg, yield 75%).

Thin-layer chromatography: (TLC) detection: R_f0.5 (petroleum ether: ethyl acetate=2: 1 expansion phase)

Mass Spectrometer Method: MS (ESI) m/z 507 (M+1)；

Nuclear magnetic resonance: 1HNMR (400MHz, CDCl₃), 1.68 (s, 9H), 0 (m, 1H), 2.28-2.45 (m, 2H), 2.90-3.25 (m, 1H), 3.40-3.80 (m, 1H), 4.05-4.25 (m, 2H), 4.65-4.90 (m, 2H), 5.31 (s, 2H), 6.30-6.40 (m, 1H), 6.60-6.70 (m, 1H), 7.05-7.15 (m, 2H), 7.20-7.25 (m, 1H), 7.30-7.40 (m, 4H), 8.41 (s, 1H).

III.3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- Base) piperidines preparation method

At 0 DEG C to tert-butyl 3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (160mg, trifluoroacetic acid (2mL) is added in methylene chloride (4mL) solution 0.32mmol), then it is warmed to room temperature reaction 4 hours, obtain 3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3, 4-d] pyrimidine -1- base) piperidines trifluoroacetate, hydrochloric acid can also be added in the solution, sulfuric acid or acetic acid, obtain corresponding hydrochloride, sulfate or acetate.It is dissolved in ethyl acetate (20mL) by the way that the crude product that solvent obtains is concentrated under reduced pressure, uses Na₂CO₃Saturated solution washs and then uses anhydrous Na₂SO₄After drying, 3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3 is obtained by the way that solvent is concentrated under reduced pressure, 4-d] pyrimidine -1- base) piperidines crude product 80mg, it is directly used in the reaction of next step, obtains corresponding hydrochloride, sulfate or acetate.

Thin-layer chromatography detection: TLC:R_f0.5 (methylene chloride: methanol=5: 1)

Mass Spectrometer Method: MS (ESI) m/z 407 (M+1)；

IV.3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- Base) piperidines is to compound 1:1- (3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- Pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone preparation method

To 3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3 at 0 DEG C, 4-d] pyrimidine -1- base) -1- piperidines (80mg, acrylic acid (23mg is sequentially added in methylene chloride (5mL) solution 0.20mmol), 0.30mmol), 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (64mg, 0.30mmol) (CAS 25952-53-8, buy producer: Shanghai Bei Zhuo Biotechnology Co., Ltd), 4- dimethylaminopyridine (DMAP) (13mg, 0.10mmol), then in room temperature reaction 2 hours, it is washed with water and then uses anhydrous Na₂SO₄After drying, by the way that the obtained crude product of solvent is concentrated under reduced pressure, by thin-layer chromatography (ethyl acetate: methanol=40: 1) purify, obtain compound 1: 1- (3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone (54mg, yield 60%).

TLC:R_f0.5 (ethyl acetate: methanol=30: 1) MS (ESI) m/z 461 (M+1)；¹HNMR (400MHz, CDCl₃) (m, 1H), 2.27-2.41 (m, 2H), 2.89-3.24 (m, 1H), 3.40-3.78 (m, 1H), 4.07-4.28 (m, 2H), 4.65-4.91 (m, 2H), 5.31 (s, 2H), 5.70-5.77 (m, 1H), 5.83 (s, 2H), 6.31-6.40 (m, 1H), 6.59-6.67 (m, 1H), 7.04-7.10 (m, 2H), 7.19-7.25 (m, 1H), 7.30-7.32 (m, 2H), 7.33-7.40 (m, 2H), 8.41 (s, 1H).

V. midbody compound and preparation method thereof

Intermediate product chemical combination 6 ' is changed according to method similar to Example 4, available such as following table compound, wherein intermediate compound 6 ' can be selected from following compound:

(4- (Phenoxymethyl) thiophene -2- base) boric acid

(4- ((5- methylphenoxy) methyl) thiophene -2- base) boric acid

(5- (Phenoxymethyl) thiene-3-yl) boric acid

(4- ((o- toloxyl) methyl) thiophene -2- base) boric acid

(4- ((m- toloxyl) methyl) thiophene -2- base) boric acid

(4- ((p- toloxyl) methyl) thiophene -2- base) boric acid

(4- ((o- methoxy phenoxy) methyl) thiophene -2- base) boric acid

(4- ((m- methoxy phenoxy) methyl) thiophene -2- base) boric acid

(4- ((p- methoxy phenoxy) methyl) thiophene -2- base) boric acid

(5- ((o- cyano-benzene oxygen) methyl) thiene-3-yl) boric acid

(5- ((p- cyano-benzene oxygen) methyl) thiene-3-yl) boric acid

(5- ((m- cyano-benzene oxygen) methyl) thiene-3-yl) boric acid

(5- ((o- methoxy phenoxy) methyl) thiene-3-yl) boric acid

(5- ((m- methoxy phenoxy) methyl) thiene-3-yl) boric acid

(5- ((pyrimidine-4-yl oxygroup) methyl) thiene-3-yl) boric acid

(5- ((2- methoxyl group -4- methylphenoxy) methyl) thiene-3-yl) boric acid

(5- ((2- methoxyl group -4- chlorophenoxy) methyl) thiene-3-yl) boric acid

(- 3 base of 5- ((2- methoxyl group -4- cyano-benzene oxygen) methyl) thiophene) boric acid

(- 3 base of 5- ((2- chloro-5-methoxyl phenoxy group) methyl) thiophene) boric acid

(- 3 base of 5- ((3- chloro-5-methoxyl phenoxy group) methyl) thiophene) boric acid

(- 3 base of 5- ((3- methoxyl group -4- cyano-benzene oxygen) methyl) thiophene) boric acid

(5- ((o- toloxyl) methyl) thiene-3-yl) boric acid

(5- ((p- toloxyl) methyl) thiene-3-yl) boric acid

(5- ((ortho-fluorophenyl oxygroup) methyl) thiene-3-yl) boric acid

(5- ((m- fluorophenoxy) methyl) thiene-3-yl) boric acid

(5- ((p- fluorophenoxy) methyl) thiene-3-yl) boric acid

((5- phenoxymethyl) thiene-3-yl) boric acid

The synthetic method of above-mentioned boronic acid compounds is similar to the preparation method that 4 compound 6 ' of embodiment is (5- (Phenoxymethyl) thiophene -2- base) boric acid.

Embodiment 5

Work as R⁵、R⁶It is H, ring 1 is benzene, L₁For singly-bound, 5 of phenyl ring and thiophene are connected, and 3 of 1H- pyrazoles [3,4-d] pyrimidine -1- base and thiophene are connected, compound 6 ' are as follows: ((5- phenyl) thiene-3-yl) boric acid

5.1Intermediate ((5- phenyl) thiene-3-yl) boric acid preparation method:

(No. CAS is 38071-58-8 to 3- bromo- 5- (phenyl) thiophene, it is purchased from Shanghai Bepharm Science & Technology Co., Ltd.) (2g, tetrahydrofuran (10mL) 8.3mmol) is dissolved in be added drop-wise under ar gas environment in tetrahydrofuran (15mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 6.7mL, 2.5mol/L).It is reacted at subzero 70 DEG C 2 hours, methyl borate (4.35g, 42mmol) then is added, the reaction was continued at subzero 70 DEG C 30 minutes, is warmed to room temperature stirring 10 minutes in the hydrochloric acid (30mL) for importing 1mol/L.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, by being concentrated under reduced pressure to give (5- (phenyl) thiene-3-yl) crude boronic acid 1.30g, since product easily decomposes, it is directly used in without purification and reacts in next step.

Thin-layer chromatography detection: TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 5)

Mass Spectrometer Method: MS (ESI) m/z 205 (M+1)；

5.2 intermediates ((5- phenyl) thiophene -2- base) boric acid and ((4- phenyl) thiophene -2- base) boric acid Preparation method

The synthetic method of ((5- phenyl) thiophene -2- base) boric acid and ((4- phenyl) thiophene -2- base) boric acid is same as the preparation method of (5- (phenyl) thiene-3-yl) boric acid.

The preparation method of the bromo- 4- of 5.32- (m- tolyl) thiophene

Work as R⁵、R⁶It is H, ring 1 is toluene, L₁For singly-bound, 5 of phenyl ring and thiophene are connected, and 3 of 1H- pyrazoles [3,4-d] pyrimidine -1- base and thiophene are connected, compound 6 ' are as follows: ((5- (3- aminomethyl phenyl)) thiene-3-yl) boric acid, the preparation method comprises the following steps:

To 2,4- dibromo thiophene (4g, 16mmol) (is purchased from West Asia reagent, product name 2,4- dibromo thiophene, goods number: CAS 3140-92-9 sequentially adds toluene boric acid (2.25g, 16mmol) in toluene solution (50mL) 1993), (No. CAS: 17933-03-8, product name 3- toluene boric acid, purchased from lark prestige science and technology, product number: 256729) Na₂CO₃(3.5g, 33mmol), Pd (PPh₃)₄, (380mg, 0.3mmol) and H₂O (50mL), then argon gas displacement protection, is warming up to 100 DEG C and reacts 12 hours, and TLC tracking reaction terminates.Reaction is finished, and is down to room temperature and is extracted with ethyl acetate, and organic liquor anhydrous Na is extracted₂SO₄After drying, by being concentrated under reduced pressure to give crude product silica gel column chromatography, eluting solvent is ethyl acetate: petroleum ether (1: 50- > 1: 20) obtains the bromo- 4- of 2- (m- tolyl) thiophene of 2g (yield 50%) white solid.

TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 30)

MS(ESI)m/z 253(M+1)；¹HNMR (400MHz, CDCl₃) 2.38 (s, 3H), 7.15-7.60 (m, 6H).

The preparation method of 5.4 intermediates (4- (3- tolyl) thiophene -2- base) boric acid

The bromo- 4- of 2- (3- tolyl) thiophene (2g, 8mmol) (such as 5.3 methods are made) is dissolved in tetrahydrofuran (10mL) and is added drop-wise in tetrahydrofuran (15mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 6.4mL, 2.5mol/L) in a nitrogen environment.It is reacted 2 hours at subzero 70 DEG C, then trimethylborate (4g is added, 40mmol), (No. CAS: 121-43-7, purchased from bass spy's reagent, product name trimethylborate, goods number: B00269101) the reaction was continued at subzero 70 DEG C 30 minutes, it is warmed to room temperature stirring 10 minutes in the hydrochloric acid (30mL) for importing 1mol/L.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, by being concentrated under reduced pressure to give (4- (3- tolyl) thiophene -2- base) crude boronic acid 1.30g, since product easily decomposes, it is directly used in without purification and reacts in next step.

TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 5)

MS(ESI)m/z 219(M+1)。

5.5 intermediates ((5- (3- methoxyphenyl)) thiene-3-yl) boric acid and ((5- (4- methoxybenzene Base)) thiene-3-yl) boric acid preparation method

The synthetic method of above-mentioned boronic acid compounds is the same as the preparation method of (4- (3- tolyl) thiophene -2- base) boric acid.

The preparation method of 5.6 intermediates (5- (m- tolyl) thiophene -2- base) boric acid

Work as R⁵、R⁶It is H, ring 1 is 3- tolyl, L₁For singly-bound, 5 of 3- tolyl and thiophene are connected, 1H- pyrazoles [3, 4-d] 2 of pyrimidine -1- base and thiophene are connected, compound 6 ' is (5- (m- tolyl) thiophene -2- base) boric acid, the preparation method comprises the following steps: to 2, 5- dibromo thiophene (4g, 16mmol) (it is purchased from Qingdao Tong Yuan Pharmaceuticals Ltd, product name 2, 5- dibromo thiophene, No. CAS: 3141-27-3) a toluene boric acid (2.25g is sequentially added in toluene solution (50mL), 16mmol) (No. CAS: 17933-03-8, purchased from Shanghai Hai Qu Chemical Co., Ltd., toluene boric acid between product name), Na₂CO₃(3.5g, 33mmol), Pd (PPh₃)₄, (380mg, 0.3mmol) and H₂O (50mL), then argon gas displacement protection, is warming up to 100 DEG C and reacts 12 hours, and TLC tracking reaction terminates.Reaction is finished, and is down to room temperature and is extracted with ethyl acetate, and organic liquor anhydrous Na is extracted₂SO₄After drying, by being concentrated under reduced pressure to give crude product silica gel column chromatography, eluting solvent is ethyl acetate: petroleum ether (1: 50- > 1: 20) obtains the bromo- 5- of 2- (m- tolyl) thiophene of 3.1g (yield 75%) white solid.

TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 30)

MS(ESI)m/z 253(M+1)；¹HNMR (400MHz, CDCl₃) 2.33 (s, 3H), 7.10-7.70 (m, 6H).

The bromo- 5- of 2- (m- tolyl) thiophene (2g, tetrahydrofuran (10mL) 8mmol) is dissolved in be added drop-wise in a nitrogen environment in tetrahydrofuran (15mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 6.4mL, 2.5mol/L).It is reacted at subzero 70 DEG C 2 hours, methyl borate (4g, 40mmol) then is added, the reaction was continued at subzero 70 DEG C 30 minutes, is warmed to room temperature stirring 10 minutes in the hydrochloric acid (30mL) for importing 1mol/L.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, by being concentrated under reduced pressure to give (5- (m- tolyl) thiophene -2- base) crude boronic acid 1.20g, since product easily decomposes, it is directly used in without purification and reacts in next step.

TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 5)

MS(ESI)m/z 219(M+1)。

5.7. preparation method similar to 5.6 intermediate

The preparation method of following intermediate is similar to 5.6 intermediate:

(5- (3,4- xylyl) thiophene -2- base) boric acid

(5- (m- anisyl) thiophene -2- base) boric acid

(5- (p- anisyl) thiophene -2- base) boric acid

(5- (4- phenyl) thiophene -2- base) boric acid

(5- (p- fluoroform phenyl) thiophene -2- base) boric acid

(5- (m- fluoroform phenyl) thiophene -2- base) boric acid

(5- (3,5- xylyl) thiophene -2- base) boric acid

(5- phenyl) thiophene -2- base) boric acid

(5- (ortho-fluorophenyl base) thiophene -2- base) boric acid

(5- (m- fluorophenyl) thiophene -2- base) boric acid

(5- (p- fluorophenyl) thiophene -2- base) boric acid

The synthetic method of above-mentioned boronic acid compounds is same as the preparation method of (5- (m- tolyl) thiophene -2- base) boric acid.

The preparation method of 5.8 intermediates (2- (styryl) thiophene -4- base) boric acid

Work as R⁵、R⁶It is H, ring 1 is styryl, L₁For singly-bound, 2 of styryl and thiophene are connected by singly-bound, 1H- pyrazoles [3,4-d] 4 of pyrimidine -1- base and thiophene are connected, m=0, ring 2 is piperidines, compound 6 ' is (2- (styryl) thiophene -4- base) boric acid, the preparation method comprises the following steps: the bromo- 2- thiophene-formaldehyde (4g of 4-, 21mmol) (CAS18791-75-8 is purchased from bass spy reagent Co., Ltd) and benzyl triphenyl phosphonium chloride phosphine (10.9g, 25mmol) are dissolved in 50mL isopropanol, then Lithium hydroxide monohydrate (1.32g, 32mmol) is added.It is warming up to 85 DEG C to react 4 hours, TLC (thin-layer chromatography detection, R_f0.5 (ethyl acetate: petroleum ether=1: 50)) tracking reaction terminate.Reaction is finished, and 50mL ethyl acetate is added, and is washed 2 times with 50mL, and separation organic layer is dry, is depressurized the crude product being spin-dried for and is dissolved in 30mL tetrahydrofuran, iodine (0.78g, 3mmol) is then added, 10h is stirred at room temperature.50mL ethyl acetate is added, successively (50mL) is washed with saturated sodium bicarbonate (50mL) and sodium sulfite, it is dry to separate organic layer, depressurize the liquid chromatogram separation for the crude product silicagel column being spin-dried for, eluting solvent is ethyl acetate: petroleum ether (1: 100- > 1: 20) obtains anti-2- (styryl) thiophene -4- bromine 4.7g, yield: 85%).

Mass Spectrometer Method: MS (ESI) m/z 265 (M+1)；

Magnetic resonance detection: 1HNMR (400MHz, CDCl₃) 6.80-7.00 (m, 2H), 7.08 (s, 1 H), 7.20 (s, 1H), 7.25-7.65 (m, 5H).

Anti- 2- (styryl) thiophene -4- bromine (2g, tetrahydrofuran (10mL) 7.5mmol) is dissolved in be added drop-wise in a nitrogen environment in tetrahydrofuran (15mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 6mL, 2.5mol/L).It is reacted at subzero 70 DEG C 2 hours, methyl borate (3.85g, 38mmol) then is added, the reaction was continued at subzero 70 DEG C 30 minutes, is warmed to room temperature stirring 10 minutes in the hydrochloric acid (30mL) for importing 1mol/L.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, by being concentrated under reduced pressure to give anti-(2- (styryl) thiophene -4- base) crude boronic acid 1.60g, since product easily decomposes, it is directly used in without purification and reacts in next step.

TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 5)

MS(ESI)m/z 231(M+1)。

The preparation method of 5.9 (2- (phenethyl) thiophene -4- base) boric acid

Work as R⁵、R⁶It is H, ring 1 is phenethyl, L₁For singly-bound, 2 of phenethyl and thiophene are connected, and 4 of 1H- pyrazoles [3,4-d] pyrimidine -1- base and thiophene are connected, and m=0, ring 2 is piperidines, and compound 6 ' is (2- (phenethyl) thiophene -4- base) boric acid.

Anti- 2- (styryl) thiophene -4- bromine (2.5g, 9.5mmol) is dissolved in ethyl acetate (30mL), and 10% palladium charcoal (1g) is added and reacts 2h, TLC (thin-layer chromatography detection, R under room temperature hydrogen environment_f0.5 (ethyl acetate: petroleum ether=1: 50)) tracking reaction terminate.Filter out the liquid chromatogram separation that the organic phase that palladium charcoal obtains passes through the crude product silicagel column being concentrated under reduced pressure to give, eluting solvent is ethyl acetate: petroleum ether (1: 100- > 1: 20) obtains 2- (phenethyl) thiophene -4- bromine 2g, yield: 80%.

Mass Spectrometer Method: MS (ESI) m/z 267 (M+1)；

Magnetic resonance detection: 1HNMR (400MHz, CDCl₃) 2.65-2.90 (m, 4H), 6.54 (s, 1H), 6.66 (s, 1H), 7.20-7.45 (m, 5H).

Crude product 1.20g is directly used in without purification and reacts in next step since product is easily decomposed.

Thin-layer chromatography TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 5)

MS(ESI)m/z 267(M+1)。

2- (phenethyl) thiophene -4- bromine (2g, tetrahydrofuran (10mL) 7.5mmol) is dissolved in be added drop-wise in a nitrogen environment in tetrahydrofuran (15mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 6mL, 2.5mol/L).It is reacted at subzero 70 DEG C 2 hours, methyl borate (3.85g, 38mmol) then is added, the reaction was continued at subzero 70 DEG C 30 minutes, is warmed to room temperature and imports 1mol/L Hydrochloric acid (30mL) in stirring 10 minutes.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, by being concentrated under reduced pressure to give (2- (phenethyl) thiophene -4- base) crude boronic acid 1.40g, since product easily decomposes, it is directly used in without purification and reacts in next step.

TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 5) MS (ESI) m/z 233 (M+1).

The compound of the present invention is prepared also by following route 3:

The reaction of step 1 is well known in reaction route 3, and in above synthetic route, the compound 5 ' as starting material is by preparing with the identical method in route 1.Compound 5 ' and compound 10 ', in organic solvent (dimethylformamide, glycol dimethyl ether, tetrahydrofuran and Isosorbide-5-Nitrae-dioxane (Isosorbide-5-Nitrae-doxane)), in Metal Palladium, such as: including tetrakis triphenylphosphine palladium Pd (PPh₃)₄, double phenyl phosphorus dichloro palladium Pd (PPh₃)₂Cl₂[1,1 '-bis- (diphenylphosphine) ferrocene] palladium chloride Pd (dppf) Cl₂In under any one or a few catalysis, reacted at 60-80 DEG C, thus carry out coupling reaction occurs to obtain compound 11 '.

The reaction of step 2 is well known in reaction route 3, and compound 11 ' is in tetrahydrofuran, in the presence of tetrabutyl ammonium fluoride, at 0 DEG C to reacting at room temperature, is thus carried out, and the reaction for carrying out de- t-butyldimethyl silane protecting group obtains compound 12 '.

The reaction of step 3 is well known in reaction route 3, and compound 12 ' sequentially adds phosphate buffer (pH=6.7), 2,2,6,6- tetramethyl piperidines-nitrogen oxides (TEMPO), NaClO in acetonitrile₂(sodium chlorite) and NaClO (sodium hypochlorite), reacts at 35 DEG C, thus carries out, and carries out hydroxyl oxidation reaction and obtains carboxylic acid compound 13 '.

The reaction of step 4 in reaction route 2 is well known, compound 13 ', in organic solvent (methylene chloride or dimethylformamide), in condensing agent 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride (EDCL) (CAS 25952-53-8, purchased from Jinan Fan Nuo Chemical Co., Ltd.), 2- (7- azo benzotriazole)-N, N, N ', N '-tetramethylurea hexafluorophosphoric acid ester (HATU) (CAS 148893-10-1, purchased from the purple chemical reagent work in Shanghai) or O- benzotriazole-N, N, N ', N '-tetramethylurea tetrafluoro boric acid ester (TBTU) (CAS 125700-67-6, purchased from Nanjing Tai Ye Biotechnology Co., Ltd) in the presence of it is anti-at room temperature with compound 14 ' It answers, thus carries out, condensation reaction occurs and obtains compound 15 '.Wherein compound 14 ' can be selected from 2-aminopyridine, 3- aminopyridine, 4-aminopyridine or aniline.

The reaction of step 5 in reaction route 3 is well known identical as the operation of step 5 in reaction route 1.The reaction of step 6 in reaction route 3 is well known identical as the operation of step 6 in reaction route 1, the starting aminopyridine being directed to is purchased from Beijing Victor Chemical Co., Ltd., wherein 3- aminopyridine article number is 462-08-8,2-aminopyridine article number is 504-29-0, and 4-aminopyridine article number is 504-24-5.

Embodiment 6

When compound 10 ' is ((5- tertiary butyl dimethyl Si methylene) thiophene -2- base) boric acid in the step 1 of reaction route 3,5 of tertiary butyl dimethyl Si methylene and thiophene are connected in compound 11 ', 1H- pyrazoles [3,4-d] 2 of pyrimidine -1- base and thiophene are connected, m=0, ring 2 is piperidines, compound 11 ' is tert-butyl -3- (4- amino -3- (5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidines -1- formic acid esters.Preparation method are as follows: at room temperature to (R) tert-butyl -3- (the iodo- 1H- pyrazoles [3 of -4- amino -3-, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (300mg, (5- (tertiary butyl dimethyl Si methylene)) thiophene -2- base is sequentially added in the solution of glycol dimethyl ether (5mL), water (2mL) 0.68mmol)) boric acid (345mg, 1.35mmol), four triphenyl phosphorus palladium (39mg, 0.03mmol), Na₂CO₃(215mg, 2.1mmol).Then it is reacted 16 hours at 80 DEG C, is cooled to room temperature, pours into 50mL water, be extracted with ethyl acetate, extract organic liquor anhydrous Na₂SO₄After drying, by the liquid chromatogram that the crude product silicagel column that solvent obtains is concentrated under reduced pressure Separation, eluting solvent is petroleum ether: ethyl acetate (8: 1 → 2: 1) obtains target product (R) tert-butyl -3- (4- amino -3- (5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidines -1- formic acid esters (275mg, yield 78%).

Thin-layer chromatography: (TLC) detection: R_f0.5 (petroleum ether: ethyl acetate=3: 1 expansion phase)

Mass Spectrometer Method: MS (ESI) m/z 545 (M+1)；

Nuclear magnetic resonance:¹HNMR (400MHz, CDCl₃): 0.20 (s, 3H), 0.21 (s, 3H), 1.59 (s, 9H), 1.69 (s, 9H), 1.60-2.45 (m, 4H), 2.90-4.90 (m, 5H), 5.31 (s, 2H), 6.30-6.40 (m, 1H), 6.60-6.70 (m, 1H), 7.00-7.15 (m, 2H), 8.40 (s, 1H).

The tert-butyl dimethyl-silicon Oxymethylene of compound 11 ' in the step 2 of reaction route 3 is connected with 5 of thiophene, 1H- pyrazoles [3, 4-d] 2 of pyrimidine -1- base and thiophene are connected, m=0, ring 2 is piperidines, compound 11 ' is (R) tert-butyl -3- (4- amino -3- (5- (tert-butyl dimethyl-silicon Oxymethylene) thiophene -2- base) -1H- pyrazoles [3, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters, compound 12 ' is (R) tert-butyl -3- (4- amino -3- (5- (hydroxy methylene) thiophene -2- base) -1H- pyrazoles [3, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters, preparation method are as follows:

At room temperature to compound 11 ', (R) tert-butyl -3- (4- amino -3- (5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidines -1- formic acid esters (250mg, 0.46mmol), tetrahydrofuran (20mL) solution in be added tetrabutyl ammonium fluoride (180mg, 0.61mmol).Then it reacts 3 hours at room temperature, is added saturated ammonium chloride solution (50mL), is extracted with ethyl acetate, extract organic liquor anhydrous Na₂SO₄After drying, by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure, eluting solvent is petroleum ether: ethyl acetate (4: 1 → 1: 1) obtains target product (R) tert-butyl -3- (4- amino -3- (5- (hydroxy methylene) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidines -1- formic acid esters (158mg, yield 80%).

Thin-layer chromatography: (TLC) detection: R_f0.5 (petroleum ether: ethyl acetate=1: 1 expansion phase)

Mass Spectrometer Method: MS (ESI) m/z 431 (M+1)；

Nuclear magnetic resonance:¹HNMR (400MHz, CDCl₃): 1.68 (s, 9H), 1.60-2.45 (m, 4H), 2.90-4.90 (m, 5H), 5.34 (s, 2H), 6.30-6.45 (m, 1H), 6.62-6.70 (m, 1H), 7.00-7.15 (m, 2H), 8.41 (s, 1H).

5 of the hydroxy methylene of compound 12 ' in the step 3 of reaction route 3 and thiophene are connected, 1H- pyrazoles [3,4-d] 2 of pyrimidine -1- base and thiophene are connected, m=0, ring 2 is piperidines, compound 12 ' is (R) tert-butyl -3- (4- amino -3- (5- (hydroxy methylene) thiophene -2- base) -1H- pyrazoles [3,4-d] Pyrimidine -1- base) piperidines -1- formic acid esters, compound 13 ' is (R) tert-butyl -3- (3- (5- formic acid thiophene -2- base) -4- amino -1H- pyrazoles [3, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters, preparation method are as follows: at room temperature to compound 12 ', (R) tert-butyl -3- (4- amino -3- (5- (hydroxy methylene) thiophene -2- base) -1H- pyrazoles [3, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (150mg, phosphate buffer (1.5mL is added in acetonitrile solution (5mL) 0.34mmol), pH=6.7), 2, 2, 6, 6- tetramethyl piperidine nitrogen oxides (5mg, 0.03mmol), NaClO₂(80mg, purity 80% are dissolved in 0.5mL water) and NaClO (0.015mL, 5% aqueous solution).Then it is reacted 5 hours at 35 DEG C, is added phosphate buffer (10mL, pH=3.6), is extracted with ethyl acetate, extract organic liquor anhydrous Na₂SO₄After drying, crude product target product (R) tert-butyl -3- (3- (5- formic acid thiophene -2- the base) -4- amino -1H- pyrazoles [3 obtained by the way that solvent is concentrated under reduced pressure, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (120mg), (compound 13 ') does not have to be further purified to be directly used in react in next step.

Thin-layer chromatography: (TLC) detection: R_f0.5 (methylene chloride: ethyl acetate=1: 2 expansion phases)

Mass Spectrometer Method: MS (ESI) m/z 445 (M+1).

Compound Compound 13 ' in the step 4 of reaction route 3 is (R) tert-butyl -3- (3- (5- formic acid thiophene -2- base) -4- amino -1H- pyrazoles [3; 4-d] pyrimidine -1- base) piperidines -1- formic acid esters; compound 14 ' is (R) -2-aminopyridine; compound 15 ' is (R) -3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3; 4-d] pyrimidine -1- base) piperidines -1- t-butyl formate, preparation method are as follows:

At room temperature to compound 13 ', tert-butyl -3- (3- (5- formic acid thiophene -2- base) -4- amino -1H- pyrazoles [3, 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (120mg, N 0.27mmol), compound 14 ' is added in dinethylformamide (5mL) solution, 2-aminopyridine (76mg, 0.81mmol) (CAS 504-29-0, purchased from AlfaAesar (China) Chemical Co., Ltd.), HATU (2- (7- azo benzotriazole)-N, N, N ', N '-tetramethylurea hexafluorophosphoric acid ester) (154mg, 0.41mmol) (No. CAS: 148893-10-1, purchased from upper sea PengShuo Biotechnology Co., Ltd) and N, N Diisopropylethylamine (172mg, 1.1mmol).Then it reacts 10 hours at room temperature, is added saturated ammonium chloride solution (20mL), is extracted with ethyl acetate, extract organic liquor anhydrous Na₂SO₄After drying; by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure; eluting solvent is petroleum ether: ethyl acetate (4: 1 → 1: 1) obtains target product compound 15 ' (R)-tert-butyl -3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3; 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (98mg, yield 70%).

Mass Spectrometer Method: MS (ESI) m/z 521 (M+1)；

Nuclear magnetic resonance:¹HNMR (400MHz, CDCl₃): 1.66 (s, 9H), 1.60-2.45 (m, 4H), 2.85-4.95 (m, 5H), 6.45-6.65 (m, 2H), 6.70-6.90 (m, 2H), 7.40-8.15 (m, 4H), 8.42 (s, 1H), 9.18 (s, 1H).Compound 16 ' in the step 5 of reaction route 3 is (R) -3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3; 4-d] pyrimidine -1- base) -1- piperidines; preparation method are as follows: at room temperature to compound 15 '; (R) tert-butyl -3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3; 4-d] pyrimidine -1- base) piperidines -1- formic acid esters (90mg, 0.17mmol) methylene chloride (5mL) solution in be added trifluoroacetic acid 2mL.Then in room temperature reaction 4 hours; (obtain (R) -3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3; 4-d] pyrimidine -1- base) -1- piperidines trifluoroacetate; hydrochloric acid, sulfuric acid can also be added in the solution, and perhaps acetic acid obtains corresponding hydrochloride, sulfate or acetate.It is dissolved in ethyl acetate (20mL) by the way that the crude product that solvent obtains is concentrated under reduced pressure, uses Na₂CO₃Saturated solution washs and then uses anhydrous Na₂SO₄After drying; compound (R) -3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3 is obtained by the way that solvent is concentrated under reduced pressure; 4-d] pyrimidine -1- base) -1- piperidines (compound 16 ') crude product 70mg, be directly used in the reaction of next step.

Mass Spectrometer Method: MS (ESI) m/z 421 (M+1).

Compound 17 ' in the step 6 of reaction route 3 is (R) -1- (3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3; 4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone, preparation method are as follows:

To (R) -3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3 at 0 DEG C; 4-d] pyrimidine -1- base) -1- piperidines (compound 16 ') (70mg; acrylic acid (23mg is sequentially added in methylene chloride (5mL) solution 0.17mmol); 0.30mmol); 1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride (64mg; 0.30mmol); 4- dimethylaminopyridine (13mg; 0.10mmol); then in room temperature reaction 2 hours, it is washed with water and then uses anhydrous Na₂SO₄After drying; the crude product obtained by the way that solvent is concentrated under reduced pressure; pass through thin-layer chromatography (ethyl acetate: methanol=40: 1) purifying; obtain compound 17 ' (R) -1- (3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3; 4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone (51mg, yield 63%).

TLC:R_f0.5 (ethyl acetate: methanol=30: 1)

MS(ESI)m/z 475(M+1)；¹HNMR (400MHz, CDCl₃)

1.60-2.45 (m, 4H), 2.89-4.95 (m, 5H), 5.25-5.65 (m, 3H), 6.55-6.95 (m, 4H), 7.30-7.32 (m, 2H), 7.30-8.15 (m, 4H), 8.42 (s, 1H), 9.14 (s, 1H).

Compound 10 ' is changed according to method similar to Example 6, available compound 51-62 as shown in table 1 below, wherein compound 10 ' can be selected from following compound:

(5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) boric acid；

(4- (tertiary butyl dimethyl Si methylene) thiophene -2- base) boric acid；

(2- (tertiary butyl dimethyl Si methylene) thiophene -4- base) boric acid

When tertiary butyl dimethyl Si methylene is connected with 5, thiophene, 2, thiophene connected with 1H- pyrazoles [3,4-d] pyrimidine radicals 3, and compound 10 ' is (5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) boric acid.

(5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) boric acid of compound 10 ' and its synthetic method Are as follows:

At room temperature to 2- bromothiophene -5- methanol (4g, 20mmol) the N of (CAS is that 79387-71-6 is purchased from the smooth Science and Technology Co., Ltd. of Haitai), successively add imidazoles (2g in dinethylformamide (50mL) solution, 30mmol) and t butyldimethylsilyl chloride (3.9g, 25mmol) (CAS 18162-48-6 is purchased from Haimen Best Fine Chemical Co., Ltd.).Then it reacts 2 hours at room temperature, 200mL water is added, then it is extracted with ethyl acetate, separation organic layer is washed 2 times with water (100mL), then organic phase is dry with anhydrous sodium sulfate, by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure, eluting solvent is ethyl acetate: petroleum ether (1: 30- > 1: 15) obtains bromo- (the 5- tertiary butyl dimethyl Si methylene) thiophene of 2-) (5.5g, yield: 90%).

Thin-layer chromatography detection: TLC:R_f0.5 (ethyl acetate: petroleum ether=1: 10) Mass Spectrometer Method: MS (ESI) m/z 307 (M+1)；

Magnetic resonance detection:¹HNMR (400MHz, CDCl₃): 0.20 (s, 3H), 0.21 (s, 3H), 1.60 (s, 9H), 5.21 (s, 2H), 6.55-6.75 (m, 2H).

Bromo- (the 5- tertiary butyl dimethyl Si methylene) thiophene (1.8g of 2-, tetrahydrofuran (15mL) 5.9mmol) is dissolved in be added drop-wise in a nitrogen environment in tetrahydrofuran (10mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 4.8mL, 2.5mol/L).It is reacted at subzero 70 DEG C 2 hours, methyl borate (3.15g, 30mmol) then is added, the reaction was continued at subzero 70 DEG C 30 minutes, is warmed to room temperature stirring 10 minutes in the hydrochloric acid (30mL) for importing 1mol/L.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, do not had to by being concentrated under reduced pressure to give compound 10 ' (5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) crude boronic acid 1.20g since product easily decomposes Purification is directly used in reacts in next step.

Mass Spectrometer Method: MS (ESI) m/z 273 (M+1)；

(4- (tertiary butyl dimethyl Si methylene) thiophene -2- base) boric acid

(2- (tertiary butyl dimethyl Si methylene) thiophene -4- base) boric acid

The synthetic method of above-mentioned boronic acid compounds is same as the preparation method of compound 10 ' (5- (tertiary butyl dimethyl Si methylene) thiophene -2- base) boric acid.

Embodiment 7

Work as R⁵、R⁶It is H, ring 1 is 2- pyridyl group, L₁For singly-bound, 5 of 2- pyridyl group and thiophene are connected, 1H- pyrazoles [3, 4-d] 2 of-pyrimidine -1- base and thiophene are connected, m=0, ring 2 is piperidines, compound 6 ' is (5- (2- pyridyl group) thiophene -2- base) boric acid, the preparation method comprises the following steps: at room temperature to 2- bromopyridine (3g, 19.1mmol) (No. CAS: 109-04-6, purchased from bass spy's reagent, article number B012654) glycol dimethyl ether (60mL) solution in sequentially add water (30mL), (thiophene -2- base) boric acid (3.4g, 26.7mmol), four triphenyl phosphorus palladium (454mg, 3.8mmol) and Na₂CO₃(6.1g, 57.3mmol).Then argon gas reacts 6 hours at 80 DEG C after replacing 3 times, is cooled to room temperature, pours into 50mL water, be extracted with ethyl acetate, and extracts organic liquor anhydrous Na₂SO₄After drying, by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure, eluting solvent is petroleum ether: ethyl acetate (100: 1 → 60: 1) obtains target product 2- (2- pyridyl group) thiophene (2.5g, yield 83%).

Thin-layer chromatography: (TLC) detection: R_f0.5 (petroleum ether: ethyl acetate=8: 1 expansion phase)；Mass Spectrometer Method: MS (ESI) m/z 162 (M+1)；Nuclear magnetic resonance:¹HNMR (400MHz, CDCl₃): 7.10-7.45 (m, 2H), 7.60-7.95 (m, 4H), 8.50-8.60 (m, 1H).

7.1 The preparation method of the bromo- 5- of 2- (2- pyridyl group) thiophene:

To the methylene chloride (100mL) of 2- (2- pyridyl group) thiophene (2.5g, 15.5mmol), methylene chloride (50mL) solution of middle dropwise addition bromine (2.50g, 15.6mmol) at 0 DEG C.It is warmed to room temperature stirring 2 hours after being added dropwise within 1 hour, pours into saturation Na₂CO₃In (200mL), organic phase is separated, anhydrous Na is used₂SO₄After drying, by the liquid chromatogram separation that the crude product silicagel column that solvent obtains is concentrated under reduced pressure, eluting solvent is petroleum ether: ethyl acetate (100: 1 → 80: 1) obtains the bromo- 5- of target product 2- (2- pyridyl group) thiophene (3g, yield 81%).

Thin-layer chromatography: (TLC) detection: R_f0.5 (petroleum ether: ethyl acetate=8: 1 expansion phase)； Mass Spectrometer Method: MS (ESI) m/z 240 (M+1)；Nuclear magnetic resonance: 1HNMR (400MHz, CDCl3): 7.15-7.50 (m, 3H), 7.65-7.95 (m, 2H), 8.55-8.60 (m, 1H).

The preparation method of (5- (2- pyridyl group) thiophene -2- base) boric acid: the bromo- 5- of 2- (2- pyridyl group) thiophene (2g, tetrahydrofuran (15mL) 8.3mmol) is dissolved in be added drop-wise in a nitrogen environment in tetrahydrofuran (15mL) solution of subzero 70 DEG C of n-BuLi (hexane solution of 6.6mL, 2.5mol/L).After 30 minutes are added dropwise, the reaction was continued at subzero 70 DEG C 2 hours, and trimethylborate (5.2g, 50mmol) then is added, the reaction was continued at subzero 70 DEG C 30 minutes, is warmed to room temperature stirring 10 minutes in the hydrochloric acid (20mL) for importing 1mol/L.It is extracted with ethyl acetate, extracts organic liquor anhydrous Na₂SO₄After drying, by being concentrated under reduced pressure to give (5- (2- pyridyl group) thiophene -2- base) crude boronic acid 1.50g, since product easily decomposes, it is directly used in without purification and reacts in next step.1 step 4-6 of following overlapping route.

(the ethyl acetate: petroleum ether=1: 5) of TLC:Rf 0.5

MS(ESI)m/z 206(M+1)。

The route 4 for preparing above-mentioned intermediate is as follows:

In the above route, R₇Selected from methyl, cyano, formamido group or acetylamino, R₇5 or 6 of pyridyl group can be located at.The thienyl boric acid replaced by selection different location, selects different location to replace bromopyridine base, the pyridyl group and thienyl of available desired link position.

7.2. preparation method similar to 7.1 intermediate

(5- (3- pyridyl group) thiophene -2- base) boric acid

5- (2- (6- picolyl)) thiophene -2- base) boric acid

5- (2- (5- picolyl)) thiophene -2- base) boric acid

(4- (2- pyridyl group) thiophene -2- base) boric acid

(4- (3- pyridyl group) thiophene -2- base) boric acid

The synthetic method of above-mentioned boronic acid compounds is similar to the preparation method of compound 6 ' (5- (2- pyridyl group) thiophene -2- base) boric acid.

7.3 work as R⁵、R⁶It is H, ring 1 is 2- (6- is cyanopyridine-based), L₁For singly-bound, 2- (6- cyano) 5 of pyridyl group and thiophene are connected, 1H- pyrazoles [3,4-d] 2 of pyrimidine -1- base and thiophene are connected, m=0, ring 2 is piperidines, compound 6 ' is (5- (6- cyanopyridine -2- base) thiophene -2- base) boric acid, the preparation method comprises the following steps: the preparation method of 5- bromo- (- 2 base of (6- cyanopyridine -2- base) thiophene) is similar to the preparation method of the bromo- 5- of 2- (2- pyridyl group) thiophene.At room temperature to bromo- ((6- cyanopyridine -2- base) thiophene -2- the base) (3g of 5-, water (30mL), bis- (pinacol combined) two boron (2.88g are sequentially added in glycol dimethyl ether (60mL) solution 11.3mmol), 11.3mmol), 1, bis- (diphenylphosphine) ferrocene palladium chloride dichloromethane complex (920mg of 1-, 1.1mmol) and NaOAc (3.1g, 22.6mmol).Then argon gas reacts 12 hours at 80 DEG C after replacing 3 times, is cooled to room temperature, pours into 50mL water, be extracted with ethyl acetate, and extracts organic liquor anhydrous Na₂SO₄After drying, 5h is stirred at room temperature in the hydrochloric acid 10mL that 30mL ethyl acetate addition 0.1M is dissolved in by the way that the crude product that solvent obtains is concentrated under reduced pressure, with saturation Na₂CO₃PH=6 or so is modulated, is extracted with ethyl acetate, organic liquor anhydrous Na is extracted₂SO₄After drying, crude product target product (5- (6- cyanopyridine -2- base) thiophene -2- base) boric acid obtained by the way that solvent is concentrated under reduced pressure is directly used in without purifying and is reacted in next step due to unstable products.

Thin-layer chromatography: (TLC) detection: Rf 0.5 (petroleum ether: ethyl acetate=6: 1 expansion phase)；Mass Spectrometer Method: MS (ESI) m/z 231 (M+1).

7.4 preparation methods similar to 7.3 intermediate

5- (2- (5- is cyanopyridine-based) thiophene -2- base) boric acid

5- (2- (5- formamido group pyridyl group) thiophene -2- base) boric acid

5- (2- (6- formamido group pyridyl group) thiophene -2- base) boric acid

5- (2- (5- (glycyl) pyridyl group) thiophene -2- base) boric acid

5- (2- (6- (glycyl) pyridyl group) thiophene -2- base) boric acid

Synthetic route of the invention is combined according to the preparation method of intermediate disclosed in embodiment 5, compound 1-50 as shown in table 1 below is obtained, synthetic route of the invention is combined according to the preparation method of intermediate disclosed in embodiment 6, has obtained compound 51-62 as shown in table 1 below:

Table 1

Embodiment 8

BTK inhibitory activity and measurement to Btk selectivity

Experimental material:

1. Bu Ludun kinase inhibitor (BTK)

Hero company, article number: PR5442A (Invitrogen-PR5442A)

2. detection kit

Sai Si biotech firm (Cisbio), article number 62TK0PEJ

3. detection plate

Platinum Elmer Co., Ltd, article number: 6007299

(PerkinElmer-6007299)

4. fluorescence plate reader/appearance and general microplate reader

Platinum Elmer Co., Ltd, article number: 2104 (PerkinElmer-2104)

Experimental procedure:

1. diluted chemical compound: untested compound and positive compound are diluted for 3 times for Buddhist nun (Ibrutinib) with dimethyl sulfoxide (DMSO) according to Shandong, totally 11 concentration, and final system concentration is from 10 μM to 0.17nM.

2. being 50mM4- hydroxyethyl piperazineethanesulfonic acid (Hepes) (pH7.5), 5mM MgCl in buffer₂, 0.01mM Na₃VO₄In the 10L reaction system of 1% bovine serum albumin (BSA), including 1nM bruton's tyrosine kinase (Btk), 1M biotinyl polypeptide (biotin-TK peptide), 20M ATP, it is 50mM 4- hydroxyethyl piperazineethanesulfonic acid (Hepes) (pH7.5) in buffer, dimethyl sulfoxide (DMSO) dilute solution of tested compounds is added in 96 hole test boards and is incubated for 90 minutes at 23 DEG C.Then 10 μ l are added and contain 20mM ethylenediamine tetra-acetic acid (EDTA), 6.7nM thymidine Btk deficiency antibody (TK) antibody, 62.5nM stop bath (SA-XL665, purchased from Shanghai Bai Li Biotechnology Co., Ltd (cisbio)), it is incubated for 60 minutes at 23 DEG C.

3. measuring fluorescence intensity of each hole at 445nm and 520nm using fluorescence plate reader/appearance and general microplate reader.The ratio of phosphorylation specification according to appended by kit, by being determined at 445nm (cumarin colour developing) relative to the colour developing ratio at 520nm (fluorescein colour developing).

4. the data that instrument is read to be calculated to the inhibiting rate of compound, IC is then calculated₅₀Value.(with mode 205 in the XLFIT5 of IDBS)

The inhibiting rate (%) of tested person compound is calculated using following formula:

Inhibition of phosphorylation rate (%)=1- { (A_C-A_X)/(A_C-A_B)}X 100

A_C: only phosphoric acid rate when addition dimethyl sulfoxide (control)

A_X: phosphoric acid rate when addition tested person compound

A_B: phosphoric acid rate when addition ATP (blank)

By the suppression curve of the inhibiting rate under each concentration based on tested person compound, tested person chemical combination is calculated Value (the IC of 50% inhibiting rate of object₅₀Value).

The measurement of the inhibitory activity of other tyrosine kinase groups such as Lck replaces Btk using various kinases, same as the above method to operate.

Inhibiting effect of the representative compound of the present invention of table 2 to tyrosine kinase

Serial number	Compound	Btk IC₅₀(nM)	Lck IC₅₀(nM)
1	Buddhist nun is replaced according to Shandong	2
2	Compound 1	28	2595
3	Compound 2	20	10000
4	Compound 3	17	3333
5	Compound 4	100	10000
6	Compound 5	11	977
7	Compound 6	54	10000
8	Compound 7	19	2924
9	Compound 8	21	3333
10	Compound 9	14	3333
11	Compound 10	21	3333
12	Compound 11	55	10000
13	Compound 12	35	10000
14	Compound 13	150	10000
15	Compound 14	55	3333
16	Compound 15	14	3200
17	Compound 16	21	10000
18	Compound 17	10	10000
19	Compound 18	4	10000
20	Compound 19	62	5095
21	Compound 20	210	5477
22	Compound 21	86	6510
23	Compound 22	157	10000
24	Compound 23	78	10000
25	Compound 24	180	7341
26	Compound 25	60	3104
27	Compound 26	68	5069
28	Compound 27	21	3333
29	Compound 28	88	4209
30	Compound 29	70	2924
31	Compound 30	64	10000
32	Compound 31	62	4095
33	Compound 32	13	5477
34	Compound 33	14	6510
35	Compound 34	92	10000
36	Compound 35	110	10000
37	Compound 36	58	3342
38	Compound 37	157	10000

39	Compound 38	73	6090
40	Compound 39	45	8090
41	Compound 40	33	10000
42	Compound 41	29	10000
43	Compound 42	30	10000
44	Compound 43	35	10000
45	Compound 44	10	10000
46	Compound 45	23	1341
47	Compound 46	8	3104
48	Compound 47	51	5069
49	Compound 48	10	7098
50	Compound 49	37	5545
51	Compound 50	13	10000
52	Compound 51	13	10000
53	Compound 52	30	5090
54	Compound 53	65	6490
55	Compound 54	80	7510
56	Compound 55	15	10000
57	Compound 56	35	10000
58	Compound 57	90	3342
59	Compound 58	112	4304
60	Compound 59	20	6090
61	Compound 60	20	8090
62	Compound 61	103	10000
63	Compound 62	4	3230
64	Compound 63	20	10000
65	Compound 64	35	> 10,000
66	Compound 65	2	2,312
67	Compound 66	10	1,613
68	Compound 67	23	1,341
69	Compound 68	14	6,510
70	Compound 69	88	4,209
71	Compound 70	8	3,104
72	Compound 71	51	5,069
73	Compound 72	75	4,289
74	Compound 73	10	7,098
75	Compound 74	37	5,545

It is above the result shows that the compound of the present invention has good selectivity inhibiting effect to Btk.

Embodiment 9

This active measurement of cell (Ramos cell) Btk specific signals conduction path of human lymphoma Rameau

1. test material

Ramos cell

2. test procedure

IgM EC₈₀Detection: collecting cell, and cell is resuspended with 1640 culture mediums containing 0.1%FBS (fetal calf serum) and adjusts concentration to 5x10⁶/mL.The hole 20L/ cell suspension is added in cell plates, is added 40L Fluo-4 carried dye (loading dye), 37 DEG C are incubated for 50 minutes.3 times of gradient dilutions IgM, final concentration of 10g/mL to 0.0046g/mL using the IgM of the horizontal screening system of high-flux cell (FLIPR) transfer 10L into cell plates, and read fluorescent value.Calculate the drug concentration (EC of IgM₈₀)。

Compound IC₅₀Detection: collecting cell, and cell is resuspended with 1640 culture mediums containing 0.1%FBS and adjusts concentration to 5x10⁶/mL.The hole 20L/ cell suspension is added in cell plates.3 times of gradient dilution untested compounds and positive compound replace Buddhist nun (Ibrutinib) according to Shandong, final concentration of 10M to 0.0046M, shift 10L compound into cell plates, 37 DEG C incubation 60 minutes.It is added 40L fluoroscopic load dyestuff (Fluo-4loading dye), 37 DEG C of incubation 50min.8 × EC of 10L is shifted using high-flux cell transfer screening system (FLIPR)₈₀IgM reads fluorescent value into cell plates.Use mapping software (Prism) GraphPad Software) production inhibiting rate curve graph, calculate compound IC₅₀。

Inhibiting effect of the representative compound of the present invention of table 3 to Ramos cell Btk specific signals conduction path

It is above the result shows that the compounds of this invention having better than the inhibiting effect for replacing Buddhist nun according to Shandong to Ramos cell Btk specific signals conduction path.

Embodiment 10

The measurement of non-Hodgkin's (Non-Hodgkin) lymphoma cell line proliferation activity

1. test material

Ramos (people's Hugh Burkitt (Burkitts) lymphoma cell)

HBL-1 (the big B lymphoma cell of people's diffusivity)

Daudi (people's Hugh Burkitt (Burkitts) lymphoma cell)

DOHH-2 (people's follicular lymphoma cell)

JeKo-1 (people's lymphoma mantle cell cell)

OCI-LY-19 (Hugh Burkitt (Burkitts) lymphoma cell)

Z-138 (people's lymphoma mantle cell cell)

SU-DHL-4 (the big B lymphoma cell of people's diffusivity)

SU-DHL-10 (the big B lymphoma cell of people's diffusivity)

WSU-DLCL2 (people's follicular lymphoma cell)

Microplate reader Molecular Devices Spectra MAX I3

Cell culture medium (RPMI1640) Sai Mofei company (Gibco) #C11875500BT

Fetal calf serum FBS hero company (Invitrogen) #10099-141

Cell Proliferation and activity detection kit (CCK-8) colleague chemical industry (Dojindo) #CK04B

384- hole plate (384- orifice plate) Corning Incorporated (Corning) #3701

2. test procedure

Cell is collected, cell is resuspended with 1640 culture mediums containing 10%FBS (fetal calf serum) and adjusts concentration to 3x10⁴/mL.50 hole μ L/ cell suspensions are added in cell plates.3 times of gradient dilution untested compounds and positive compound replace Buddhist nun (Ibrutinib) according to Shandong, shift 5 μ L compound solutions into cell plates, Make final concentration from 50 μM or 1 μM to 0.128nM or 0.0026nM, 37 DEG C are incubated for 72 hours.2- (2- methoxyl group -4- nitrobenzophenone) -3- (4- nitrobenzophenone) -5- (2 is added, 4- disulfonic acid benzene) -2H- tetrazolium monosodium salt solution, i.e. CCK-8 solution (is purchased from Shanghai previous existence Biotechnology Co., Ltd, product name CCK-8 kit, article number 40203ES60) 5 μ L, 37 DEG C are incubated for 3 hours.Fluorescent value is read using microplate reader.Cell Proliferation curve graph is made using mapping software Prism5.0 (GraphPad Software), calculates compound IC₅₀。

Inhibitory rate of cell growth (%)=1- { (AC-AX)/(AC-AB) } X 100

AC: only negative control absorbance value when addition dimethyl sulfoxide (control)

AX: the absorbance value of addition tested person compound well

AB: the absorbance value of blank control (blank)

Inhibiting effect of the representative compound of the present invention of table 4 to non-Hodgkin lymphoma cell Proliferation

Serial number	Compound	Romas IC₅₀(M)	HBL-1IC₅₀(M)	Daudi IC₅₀(M)
1	Buddhist nun is replaced according to Shandong	1.62	15.4	4
2	Compound 1	11.3	10.5	20
3	Compound 3	20	32.7	32
4	Compound 17	0.69	1.45	0.51
5	Compound 18	4.90	11	20
6	Compound 30	6.34	0.22	3.80
7	Compound 32	8	5.57	11
8	Compound 33	3.57	0.43	3.28
9	Compound 44	4.45	8.25	62
10	Compound 45	4.70	0.70	1.86
11	Compound 38	0.47	0.60	2.10
12	Compound 39	5.45	0.07	1.50
13	Compound 40	6.80	0.14	3.80
14	Compound 51	30	7.6	12.15
15	Compound 54	0.65	0.79	0.80
16	Compound 63	0.21	0.005	0.25
17	Compound 64	0.30	0.02	0.15
18	Compound 65	1.03	1.25	10.15
19	Compound 66	0.65	0.79	0.80
20	Compound 67	0.21	0.005	0.25
21	Compound 68	0.30	0.02	0.15
22	Compound 69	1.03	1.25	10.15
23	Compound 70	1.03	1.25	10.15
24	Compound 71	1.03	1.25	10.15
25	Compound 73	0.65	0.79	0.80

Table 5

It is above the result shows that the compound of the present invention has apparent inhibiting effect to non-Hodgkin's (Non-Hodgkin) lymphoma cell proliferation, and part of compounds activity is substantially better than according to Shandong for Buddhist nun.

Embodiment 11

The compounds of this invention induces the pharmacodynamic study in arthritis mouse model in II Collagen Type VI

Collagen-induced Arthritis is that have the immune rear experimental animal model induced of species specificity collagen I I type.Become the animal model that research rheumatoid arthritis is ideal at present because its genetic background and immunopathology change and clinical rheumatoid arthritis is very much like.

Model production method: DBA/1J mouse, 7 week old, weight 18-22g, male, (being purchased from Jinan Ao Nuo bioengineering Co., Ltd, product name DBA/1J mouse, model: DBA/1J).Take appropriate ox II Collagen Type VI, (4mg collagen/ml) is dissolved in 0.01mol/L acetic acid, it is fully emulsified under ice bath environment with equivalent complete Freund's adjuvant, every mouse is with 0.1ml (200 μ g containing collagen) emulsion, make intracutaneous injection in root of the tail portion, it is emulsified again with equivalent collagen through incomplete Freund's adjuvant within 21st day, booster immunization 1 time.

Material and method: test-compound 17, compound 63, compound 64 are dissolved in polyethylene glycol 400 with concentration 200mg/ml: the ethoxylate (KolliphorRH40)=8: 2 of castor oil.(the ethoxylate CAS 61788-85-0 of castor oil is purchased from BASF Aktiengesellschaft) feeds II Collagen Type VI by the dosage of 25mg/kg by oral administration once a day and induces arthritic DBA/1J mouse.Mouse is divided into four test-compound 17, compound 63, compound 64 and solvent groups, and wherein solvent group passes through ethylene glycol 400: ethoxylate=8 of castor oil: 2 ratio, is used once a day to mouse with the dosage of 100mg/kg Medicine, continuous use 14 days.

Observation index and analysis: arthritis index scores.Make arthritis index scoring by the arthritis score standard of WoodShi.0 point, normally；1 point, redness is related to 1 finger joint；2 points, redness is related to 2 or more articulations digitorum manus or entire sufficient pawl mild redness；3 points, sufficient pawl redness is heavier；4 points, sufficient pawl severe is red and swollen, and anchylosis lacks flexibility.The damage of every pawl is all divided into the total mark that 0-4 calculates four limbs in 4 sufficient pawls, it is expressed as a percentage that arthritic limbs number occurs for every group of mouse, and compare the integral (arthritis index) of different time, arthritic disease incidence and arthritic onset time are also recorded simultaneously.See Fig. 1, by Fig. 1 as it can be seen that as administration time increases, the arthritic conditions of mouse make moderate progress.

Growth in vitro inhibitory action of 12 compound of embodiment to chronic leukemia K562 cell

The external CellTiter- of chronic leukemia K562 cellATP luminescent cell viability examination

K562 leukemic cells maintain 37 DEG C from ATCC, 5%CO₂Atmosphere and in Dulbecco culture medium (IMDM) and 10% fetal calf serum.Cell is with 6*10³The density in a/hole is seeded on 96 orifice plates, test compound is dissolved in DMSO, and it is 0 μM, 0.3 μM, 0.5 μM, 1 μM, 2 μM, 3 μM, 5 μM, 10 μM, 20 μM, 30 μM, 50 μM and 100 μM with concentration and acts on K562 cell 72 hours, then it is detected with CellTiter-Glo luminescence method cell viability detection kit by the cell after compound effects, and records luminous value.

Part of compounds In Leukemic Cells In Vitro K562 cell viability of the present invention, which influences result, to be enumerated in the following table 6.

Table 6

Claims

It is a kind of with logical formula (I) compound represented, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide:

Wherein: W is nitrogenous saturated heterocyclyl, the benzene methylene (C of 4-6 member₃-C₆) naphthenic base or [3.3-5] miscellaneous loop coil base containing nitrification；

R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；Substituted-phenyl replaces the substituent group on nitrogen-containing hetero phenyl that can be each independently selected from halogen, (C₁-C₄) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄) one of halogenated alkoxy or a variety of；

Any one integer between n=0-4；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 1, such as formula (II) institute Show:

Wherein, W is nitrogenous saturated heterocyclyl, the benzene methylene (C of 4-6 member₃-C₆) naphthenic base or [3.3-5] miscellaneous loop coil base containing nitrification；

R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 2, as shown in formula (III):

Wherein, R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 1 or 3, as shown in formula (IV):

Wherein, the R²Selected from phenyl, substituted-phenyl, benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkoxy, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkane Base, benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 4, as shown in formula (V):

Wherein, X, Y are each independently selected from CH or N, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 5, as shown in formula (VI):

Wherein, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 5, as shown in formula (VII):

Wherein, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 5, as shown in formula (VIII):

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 4, as shown in formula (IX):

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 4, such as formula (X) It is shown:

Wherein, X, Y and Z are each independently selected from CH or N；R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄One or more of) halogenated alkoxy.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 1 or 3, as shown in formula (XI):

Wherein, R¹Selected from phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkoxy, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero benzene The base, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 11, as shown in formula (XII):

Wherein, X, Y are each independently selected from CH or N, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 11, as shown in formula (XIII):

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 11, as shown in formula (XIV):

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
The mixing of compound, its cis-trans-isomer, cis-trans-isomer according to claim 11 Object, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (XV):

Wherein, X, Y and Z are each independently selected from CH or N；R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 12, as shown in formula (XVI):

Wherein, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 12, as shown in formula (XVII):

Wherein, R⁴Selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl, amine formyl, acetamido and (C₁-C₄One of) halogenated alkoxy.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 1, as shown in formula (XVIII):

Wherein, R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, substituted benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

Any one integer between n=0-4；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 18, as shown in formula (XIX):

Wherein, R²Selected from phenyl, substituted-phenyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and takes For one or more of nitrogen-containing hetero phenyl；

R¹Selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl or (C₁-C₄) halogenated alkoxy；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
According to compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or the N- oxide described in claim 19, as shown in formula (XX):

Wherein, R²Selected from phenyl, substituted-phenyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；

R³Selected from (C₂-C₄) alkenyl, (C₂-C₄) alkynyl, amine acrylic, N, N- bis- replaces amine acrylic and (C₄-C₇) one or more of the acrylic that replaces of nitrogenous saturated heterocyclic, wherein the substituent group replaced on amine includes (C₁-C₄) one or more of alkyl and hydroxyl.
The mixing of compound, its cis-trans-isomer, cis-trans-isomer according to claim 20 Object, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide, as shown in formula (XXI):

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 14, as shown in formula (X XII)

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 18, as shown in formula (XXIII):

Wherein, R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄) one of halogenated alkoxy or a variety of.
Compound, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 18, as shown in formula (XXIV):

Wherein, X, Y and Z are each independently selected from CH or N；R⁵、R⁶It is each independently selected from H, halogen, (C₁-C₃) alkoxy, (C₁-C₄) alkyl, cyano, (C₁-C₄) halogenated alkyl and (C₁-C₄One or more of) halogenated alkoxy.
Compound described in any one of -24, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide according to claim 1, wherein the compound is selected from one or more of following compound:

1- (3- (4- amino -3- (5- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- tolylthiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- methyl -4- (Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- (Phenoxymethyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((2- methyl) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- Base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((m- methyl) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((p- methyl) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((2- methoxyl group) phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((3- methoxyl group) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (4- ((4- methoxyl group) Phenoxymethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- ((2- cyano) Phenoxymethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- ((4- cyano) Phenoxymethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- ((3- cyano) Phenoxymethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- (2- methoxybenzene) oxygen methylthiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (2- (3- methoxybenzene) oxygen methylthiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- benzene thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (Phenoxymethyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((pyrimidine-4-yl oxygroup) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- ((2- methoxyl group -4- methylphenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (4- amino -3- (5- ((2- methoxyl group -4- chlorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] Pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((2- methoxyl group -4- cyano-benzene oxygen) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((2- chloro-5-methoxyl phenoxy group) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((3- chloro-5-methoxyl phenoxy group) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((3- methoxyl group -4- cyano-benzene oxygen) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (2- methylenedioxy phenoxy methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- toloxyl methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- aminomethyl phenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3,4- 3,5-dimethylphenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- methoxyphenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- methoxyphenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- tolylthiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- tolylthiophene -3- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- trifluoromethyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- benzotrifluoride) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3,5- dimethylbenzene) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperazine Pyridine -1- base) propyl- 2- alkene -1- ketone；

(S) -1- (3- (4- amino -3- (5- tolylthiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (2- fluorophenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- fluorophenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- fluorophenyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (3- aminomethyl phenyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (4- methoxyphenyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- methoxyphenyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((2- fluorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((3- fluorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- ((4- fluorophenoxy) methyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(S) -1- (3- (4- amino -3- (5- (phenoxymethyl) thiene-3-yl) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (phenethyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) trans- 1- (3- (4- amino -3- (2- styrene thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

1- (3- (1- (4- amino -3- (2- (3- methylbenzene) oxygen methyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone

(R) -1- (3- (4- amino -3- (5- (2- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine - 1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (3- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (4- pyridine amine formyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (anilino- formoxyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (2- pyridine amido formacyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (3- pyridine amido formacyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (4- pyridine amido formacyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (2- (anilino- formoxyl) thiophene -4- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (2- pyridine amido formacyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (3- pyridine amido formacyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (4- pyridine amido formacyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (anilino- formoxyl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (pyridine -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (pyridin-3-yl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (pyridine -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (4- (pyridin-3-yl) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperazine Pyridine -1- base) propyl- 2- alkene -1- ketone；

(S) -1- (3- (4- amino -3- (5- (pyridine -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (6- cyanopyridine -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base)-picolinamide；

(R) -6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base)-Niacinamide；

(R)-N- (6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base) pyridine -2- base) acetamide；

(R)-N- (6- (5- (1- (1- Antiepilepsirin -3- base) -4- amino -1H- pyrazoles [3,4-d] pyrimidin-3-yl) thiophene -2- base) pyridin-3-yl) acetamide；

(R) -1- (3- (4- amino -3- (5- (6- picoline -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone；

(R) -1- (3- (4- amino -3- (5- (5- picoline -2- base) thiophene -2- base) -1H- pyrazoles [3,4-d] pyrimidine -1- base) piperidin-1-yl) propyl- 2- alkene -1- ketone.
A kind of such as salt of following formula (XXV) compound represented or the compound,

Wherein, R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkene Base, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；R⁷Selected from H orWherein R³Selected from one of trifluoromethyl, tert-butoxy and benzyloxy；Work as R⁷When for H, the salt of formula (XXV) compound can be one of hydrochloride, sulfate, acetate and trifluoroacetate.
According to claim 26 compound represented, wherein the compound is as follows:

Wherein, R¹Or R²It is each independently selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；R⁷Selected from H orWherein R³Selected from trifluoromethyl, one of tert-butoxy and benzyloxy；Work as R⁷When for H, the salt of formula (XXVI) compound can be one of hydrochloride, sulfate, acetate and trifluoroacetate.
According to claim 26 compound represented, wherein the compound is as follows:

Wherein, R₁Selected from H, (C₁-C₄) alkyl, (C₁-C₄) alkoxy, (C₁-C₄) halogenated alkyl, (C₁-C₄) halogenated alkoxy, phenyl, substituted-phenyl, benzene (C₂-C₄) alkynyl, benzene (C₁-C₄) alkyl, benzene methylene (C₃-C₆) naphthenic base, substituted benzene (C₁-C₄) alkyl, benzene oxyalkyl, substituted benzene oxygen alkyl, benzene (C₁-C₄) alkoxy, substituted benzene (C₁-C₄) alkoxy, benzene (C₂-C₄) alkenyl, substituted benzene (C₂-C₄) alkenyl, nitrogen-containing hetero the phenyl, (C for replacing nitrogen-containing hetero phenyl, nitrogen-containing hetero phenyl to replace₁-C₄) alkyl, nitrogen-containing hetero phenyl replace (C₁-C₄) alkoxy andOne or more of；Wherein R " is selected from one or more of phenyl, substituted-phenyl, nitrogen-containing hetero phenyl and substitution nitrogen-containing hetero phenyl；R⁷Selected from H orWherein R³Selected from one of trifluoromethyl, tert-butoxy and benzyloxy；Work as R⁷When for H, the salt of formula (XXVII) compound can be one of its hydrochloride, sulfate, acetate and trifluoroacetate.
A kind of such as following formula (XXVIII) compound represented, wherein the compound is as follows:

Wherein, R⁸Selected from H, Br or boronate, R⁹Selected from H, methyl, cyano, amine formyl or acetamido.
Compound according to claim 29, wherein shown in the compound such as following formula (XXIX):

Wherein, R⁸Selected from H, Br or boronate, R⁹Selected from H, methyl, cyano, amine formyl or acetamido.
Compound according to claim 29, wherein shown in the compound such as following formula (XXX):

Wherein, R⁸Selected from H, Br or boronate, R⁹Selected from H, methyl, cyano, amine formyl or acetamido.
Compound according to claim 29, wherein shown in the compound such as following formula (XXXI):

Wherein, R⁸Selected from H, Br or boronate, R⁹Selected from H, methyl, cyano, amine formyl or acetamido.
Compound according to claim 29, wherein shown in the compound such as following formula (XXXII):

Wherein, R⁸Selected from H, Br or boronate, R⁹Selected from H, methyl, cyano, amine formyl or acetamido.
Compound described in any one of claim 1-25, its cis-trans-isomer, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide are in preparation for treating and including the purposes in malignant tumour, autoimmune disease and anaphylactia related drugs.
Purposes according to claim 34, wherein the malignant tumour includes one or more of lymthoma, plasmacytoma and leukaemia.
Purposes according to claim 36, wherein the lymthoma includes one or more of non-Hodgkin lymphoma, follicular lymphoma, lymphoma mantle cell, small lymphocyte lymthoma, mantle cell lymphoma, intravascular maxicell type B cell lymphoma, Burkitt lymphoma, AIDS related lymphoma and marginal zone B-cell lymphoma；Preferably, the non-Hodgkin lymphoma includes B cell non-Hodgkin lymphoma；It is highly preferred that the B cell non-Hodgkin lymphoma includes one or more of diffusivity large B cell lymphoid tumor and human B lymphocytes' tumor.
Purposes according to claim 34, wherein the autoimmune disease includes one or more of arthritis, rheumatism, inflammatory enteritis and lupus erythematosus.
A kind of bruton's tyrosine kinase inhibitor combination, including compound, its cis-trans-isomer described in any one of the claims in the present invention 1-25, the mixture of cis-trans-isomer, optical enantiomorphs, the mixture of enantiomter, racemic modification or N- oxide.