CN106146511A

CN106146511A - Pyrazolopyrimidine derivative, preparation method, pharmaceutical composition and purposes

Info

Publication number: CN106146511A
Application number: CN201510158304.5A
Authority: CN
Inventors: 洪健; 许忻; 乐小勇
Original assignee: An Run Pharmaceutical Technology (suzhou) Co Ltd
Current assignee: An Run Pharmaceutical Technology (suzhou) Co Ltd
Priority date: 2015-04-03
Filing date: 2015-04-03
Publication date: 2016-11-23

Abstract

The invention discloses Pyrazolopyrimidine derivative, preparation method, pharmaceutical composition and purposes.The invention provides a kind of Pyrazolopyrimidine derivative shown in formula I, its stereoisomer, solvate, pharmaceutically acceptable salt, active metabolite or prodrug.The Pyrazolopyrimidine derivative shown in formula I of the present invention has good inhibitory activity, the especially growth to tumor cell to bruton's tyrosine kinase (Btk) and has good vivo and vitro inhibitory activity, has good market-oriented prospect.

Description

Pyrazolopyrimidine derivative, preparation method, pharmaceutical composition and purposes

Technical field

Present invention relates particularly to Pyrazolopyrimidine derivative, preparation method, pharmaceutical composition and purposes.

Background technology

Bruton's tyrosine kinase (Bruton ' s tyrosine kinase, Btk), a kind of nonreceptor tyrosine kinase Tec family Member, is the key signal expressed in all hematopoetic cell types in addition to T lymphocyte and natural killer cell Enzyme.Btk stimulates thin to the B of response in downstream cellular in connection cell surface B-cell receptor (B-cell receptor, BCR) Born of the same parents' signal transduction path is played the part of vital role.

Btk be B cell grow, activate, signal conduction and survival key regulators (Kurosaki, Curr Op Imm, 2000,276-281；Schaeffer and Schwartzberg, Curr Op Imm 2000,282-288).It addition, Btk exists Other hematopoietic cell signal transduction paths numerous work, such as Toll-like receptor (the Toll in macrophage Likereceptor, TLR) and the TNF-α of cytokine receptor mediation produces, IgE in mastocyte Receptor (Fc ε RI) signal conduction, in B-pedigree lymphoid cell suppress Fas/APO-1 apoptotic signal conduction with And the platelet aggregation of collagen stimulation.See for example (2003) such as C.A.Jeffries, Journal of Biological Chemistry 278:26258-26264；N.J.Horwood etc. (2003), The Journal of Experimental Medicine197: 1603-1611；Iwaki etc. (2005), Journal of Biological Chemistry280 (48): 40261-40270； Vassilev etc. (1999), Journal of Biological Chemistry274 (3): 1646-1656；Quek etc. (1998), Current Biology 8 (20): 1137-1140.

BTK targeted inhibitors has started to for oncotherapy at present, but its inhibitory activity and drug effect are notable not enough, face Need on Chuan use larger dose, common side reaction include thrombocytopenia, diarrhoea, middle granulocytopenia, anemia and Upper respiratory tract infection etc., thus need nonetheless remain for researching and developing further activity or pharmacological characteristics than more preferable compound, as That update, that improve or more efficient BTK acceptor inhibitor such that it is able to gain more insight into such medicine and BTK Relation between target protein and play its Antitumor Mechanism, clinical therapy of tumor is all had very important by these Meaning.

Summary of the invention

The technical problem to be solved is to overcome the biology of bruton's tyrosine kinase inhibitor in prior art Activity and drug effect not substantially, using dosage provide Pyrazolopyrimidine derivative, system compared with the defect such as big, side effect is big Preparation Method, pharmaceutical composition and purposes.The Pyrazolopyrimidine derivative shown in formula I of the present invention is to bruton's tyrosine Kinases (BTK) has good inhibitory activity, the especially growth to tumor cell and has good vivo and vitro suppression work Property, there is good market-oriented prospect.

The present inventor is found by further investigation, and it is irreversible that the Pyrazolopyrimidine derivative of Formulas I structure has effective Btk Inhibitory action.

The invention provides a kind of Pyrazolopyrimidine derivative shown in formula I, its stereoisomer, solvate, medicine Acceptable salt, active metabolite or prodrug on,

Wherein: L be O, S,When L be O, S, Z isWhen L isTime, Z is Wherein, X is halogen atom (such as fluorine atom, chlorine atom or bromine atoms), cyano group, R is C₁～C₄Alkyl (described " C₁～C₄Alkyl " such as methyl, ethyl, propyl group, Isopropyl, butyl, isobutyl group or the tert-butyl group, preferably methyl)；

Ar is substituted or unsubstituted aryl (preferably " substituted or unsubstituted C₅～C₁₀Aryl "；Described " replaces Or unsubstituted C₅～C₁₀Aryl " preferred substituted or unsubstituted phenyl；Described " substituted phenyl " can be quilt The substituted phenyl of one or more fluorine atoms, by one or more methyl substituted phenyl, replaced by one or more cyano group Phenyl, the phenyl replaced by one or more trifluoromethyls, by the phenyl of one or more methoxy substitutions, by one Or the phenyl of multiple vinyl substituted, the phenyl that replaced by one or more acetenyls, or taken by one or more phenyl The phenyl in generation；Described " phenyl replaced by one or more fluorine atoms " such as OrDescribed " by one or more methyl substituted phenyl " such as 4-aminomethyl phenyl；Described " by one Or the multiple substituted phenyl of cyano group " such as 2-cyano-phenyl；Described " phenyl replaced by one or more trifluoromethyls " example Such as 4-trifluoromethyl；Described " by the phenyl of one or more methoxy substitutions " such as 3-methoxyphenyl；Described " by the phenyl of one or more vinyl substituted " such asDescribed " is taken by one or more acetenyls The phenyl in generation " such asDescribed " phenyl being optionally substituted with one or more phenyl groups " such as) or substituted or unsubstituted heteroaryl (described " substituted or unsubstituted heteroaryl " is preferred Hetero atom be nitrogen-atoms, hetero atom number be the substituted or unsubstituted C of 1-2₃～C₉Heteroaryl；Described is " miscellaneous former The substituted or unsubstituted C that son is nitrogen-atoms, hetero atom number is 1-2₃～C₉Heteroaryl " preferably hetero atom is that nitrogen is former Son, hetero atom number are the substituted or unsubstituted C of 1₃～C₅Heteroaryl, it is described that " hetero atom is nitrogen-atoms, miscellaneous Atomic number is the substituted or unsubstituted C of 1₃～C₅Heteroaryl " preferred substituted or unsubstituted pyridine radicals；Described " unsubstituted pyridine radicals " such as)；Described " substituted or unsubstituted aryl or substituted or unsubstituted miscellaneous Aryl " described in " replacement " refer to that (such as fluorine atom, chlorine atom or bromine are former selected from halogen atom by one or more Son), cyano group, substituted or unsubstituted alkyl (the most substituted or unsubstituted C₁～C₄Alkyl, described " does not takes The alkyl C in generation₁～C₄Alkyl " preferably methyl, described " substituted C₁～C₄Alkyl " preferred trifluoromethyl), Alkoxyl (preferably C₁～C₄Alkoxyl, described " C₁～C₄Alkoxyl " preferred methoxyl group), thiazolinyl (preferably C₂～C₄Thiazolinyl, described " C₂～C₄Thiazolinyl " preferred vinyl), alkynyl (preferably C₂～C₄Alkynyl, institute " the C stated₂～C₄Alkynyl " preferred acetenyl) and aryl (preferably C₅～C₁₀Aryl, described " C₅～C₁₀'s Aryl " preferred phenyl) substituent group replaced, described " replacement " described in " substituted or unsubstituted alkyl " Refer to be replaced by one or more halogen atoms (such as fluorine atom, chlorine atom or bromine atoms) that (described is " substituted Alkyl " preferred trifluoromethyl), when there is multiple substituent group, described substituent group can be identical or different；

V is hydrogen or fluorine atom；

Y is substituted or unsubstituted alkylidene (the most substituted or unsubstituted C₁～C₆Alkylidene, described " replace Or unsubstituted C₁～C₆Alkylidene " preferred substituted or unsubstituted methylene, described " substituted methylene " is excellent Select the substituted methylene of pyrrolidinyl；Described " the substituted methylene of pyrrolidinyl " such as)、 Cycloalkyl (preferably C₄～C₇Cycloalkyl, described " C₄～C₇Cycloalkyl " preferably C₅～C₆Cycloalkyl；Institute " the C stated₆Cycloalkyl " such as cyclohexyl) or Heterocyclylalkyl (preferably hetero atom is oxygen, sulfur or nitrogen-atoms, hetero atom The C that number is 1-3₄～C₇Heterocyclylalkyl, described " hetero atom be oxygen, sulfur or nitrogen-atoms, hetero atom number be 1-3 Individual C₄～C₇Heterocyclylalkyl " preferably hetero atom be nitrogen-atoms, hetero atom number be the C of 1-2₄～C₅Heterocyclylalkyl； Described " hetero atom be nitrogen-atoms, hetero atom number be the C of 1-2₄～C₅Heterocyclylalkyl " preferably pyrrolidinyl or Piperidyl；Described " pyrrolidinyl " is preferredDescribed " piperidyl " is preferred)； The described replacement described in " substituted or unsubstituted alkylidene " refers to by " hetero atom is oxygen, sulfur or nitrogen-atoms, miscellaneous Atomic number is the C of 1-3₄～C₇Heterocyclylalkyl " (preferably hetero atom be nitrogen-atoms, hetero atom number be the C of 1-2₄~ C₅Heterocyclylalkyl, described " hetero atom be nitrogen-atoms, hetero atom number be the C of 1-2₄～C₅Heterocyclylalkyl " be Pyrrolidinyl), C₁～C₆Alkoxyl, C₁～C₆Alkylthio group and C₁～C₆Alkylamino in one or more taken Generation.

In the present invention, the preferred O of described L, S,Further preferably O, S and

In the present invention, the preferred substituted or unsubstituted aryl of described Ar, described " substituted or unsubstituted aryl " is excellent Choosing " substituted or unsubstituted C₅～C₁₀Aryl "；Described " substituted or unsubstituted C₅～C₁₀Aryl " preferably Substituted or unsubstituted phenyl；Described " substituted phenyl " preferably by one or more selected from fluorine atom, chlorine atom, Bromine atoms, trifluoromethyl, cyano group, methyl, methoxyl group, vinylAcetenylWith The substituent group of phenyl is replaced；" phenyl replaced by one or more fluorine atoms " such as " by one or more methyl substituted phenyl " such as 4-aminomethyl phenyl；" by one or The substituted phenyl of multiple cyano group " such as 2-cyano-phenyl；" phenyl replaced by one or more trifluoromethyls " such as 4-trifluoro Aminomethyl phenyl；" by the phenyl of one or more methoxy substitutions " such as 3-methoxyphenyl；" by one or more vinyls Substituted phenyl " such as" phenyl replaced by one or more acetenyls " such as " phenyl being optionally substituted with one or more phenyl groups " such as

In the present invention, described V is preferably hydrogen.

In the present invention, described Y preferred substituted or unsubstituted alkylidene (the most substituted or unsubstituted C₁～C₆Asia Alkyl, described " substituted or unsubstituted C₁～C₆Alkylidene " preferred substituted or unsubstituted methylene, described " substituted methylene " preferably substituted methylene of pyrrolidinyl；Described " the substituted methylene of pyrrolidinyl " such as) or Heterocyclylalkyl (preferably hetero atom be oxygen, sulfur or nitrogen-atoms, hetero atom number be 1-3 C₄～C₇Heterocyclylalkyl, described " hetero atom be oxygen, sulfur or nitrogen-atoms, hetero atom number be the C of 1-3₄～C₇'s Heterocyclylalkyl " preferably hetero atom be nitrogen-atoms, hetero atom number be the C of 1-2₄～C₅Heterocyclylalkyl；Described is miscellaneous former The C that son is nitrogen-atoms, hetero atom number is 1-2₄～C₅The preferred pyrrolidinyl of Heterocyclylalkyl or piperidyl；Described " pyrrolidinyl " is preferredDescribed " piperidyl " is preferred)。

In the present invention, when L be O, S,Time, Z is preferably Wherein, X preferred Cl, Br or CN, the preferred pyrrolidinyl of Y or piperidyl；Described " pyrrolidine Base " preferablyDescribed " piperidyl " is preferred)。

In the present invention, when L isTime, Z is preferably Wherein, X preferred Cl, Br or CN, Y is excellent Select pyrrolidinyl or piperidyl；Described " pyrrolidinyl " is preferredDescribed " piperidyl " is excellent Choosing)。

In the present invention, described Pyrazolopyrimidine derivative shown in formula I, preferably such as the compound of Formulas I-A:

Wherein, the definition of Ar, L, Y, Z and V is the most same as above.

In the present invention, described Pyrazolopyrimidine derivative shown in formula I, further preferred following arbitrary compound:

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine third -2-alkene-1-ketone (compound 1)；

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-chlorine third -2-alkene-1-ketone (compound 2)；

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-fluorine third -2-alkene-1-ketone (compound 3)；

(R, E)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-4,4, 4-tri-fluoro-but-2-ene-1-ketone (compound 4)；

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-fluoroform Base acrylate-2-alkene-1-ketone (compound 5)；

(R, Z)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-4-oxo But-2-ene cyanogen (compound 6)；

(R, E)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-4-oxo But-2-ene acid methyl ester (compound 7)；

(R, E)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-3-chlorine third -2-alkene-1-ketone (compound 8)；

(R, Z)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-3-bromine third -2-alkene-1-ketone (compound 9)；

(R, E)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-3-bromine third -2-alkene-1-ketone (compound 10)；

(R, E)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-4-oxo But-2-enamides (compound 11)；

(R)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-2-fluorine acrylate-2-alkene-1-ketone (compound 12)；

(R)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-2-bromine the third 2-alkene-1-ketone (compound 13)；

(R)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-2-chlorine acrylate-2-alkene-1-ketone (compound 14)；

(R, E)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-4,4,4-trifluoro but-2-ene-1-ketone (compound 15)；

(R, E)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-4-oxo but-2-ene acid methyl ester (compound 16)；

(R, Z)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-4-oxo but-2-ene cyanogen (compound 17)；

(S)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-2-fluorine acrylate-2-alkene-1-ketone (compound 18)；

(S)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-2-chlorine acrylate-2-alkene-1-ketone (compound 19)；

(S)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-2-bromine acrylate-2 alkene-1-ketone (compound 20)；

(S, E)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-4,4,4-trifluoro but-2-ene-1-ketone (compound 21)；

(S, E)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-4-oxo but-2-ene acid methyl ester (compound 22)；

(S, Z)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-4-oxo but-2-ene cyanogen (compound 23)；

(S, E)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] nafoxidine-1- Base]-4-(dimethylamino) but-2-ene-1-ketone (compound 24)；

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl] acrylate-2-alkene-1- Ketone (compound 25)；

(R)-[4-[4-amino-1-(1-(vinyl sulfone(RemzaolHuo Xingranliaohuoxingjituan)) piperidines-3-base)-1H-pyrazolo [3,4-d] pyrimidin-3-yl] phenyl] (phenyl) first Ketone (compound 26)；

(R, E)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-4-oxo fourth -2-alkene cyanogen (compound 27)；

(R)-1-[3-[4-amino-3-(4-(difluoro (phenyl) methyl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] acrylate-2-alkene-1-ketone (compound 28)；

(R)-3-[4-(difluoro (phenyl) methyl) phenyl]-1-(1-(vinyl sulfone(RemzaolHuo Xingranliaohuoxingjituan)) piperidines-3-base)-1H-pyrazolo [3,4-d] pyrimidine-4- Ammonia (compound 29)；

[(R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) piperidin-1-yl] ((R)-epoxy second Alkane-2-base) ketone (compound 30)；

[(R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) piperidin-1-yl] ((S)-epoxy second Alkane-2-base) ketone (compound 31)；

(R)-1-[3-[4-amino-3-(4-Phenylsulfanylphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine acrylate-2- Alkene-1-ketone (compound 32)；

(R)-1-[3-[4-amino-3-(4-benzyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine acrylate-2-alkene -1-ketone (compound 33)；

(R)-1-[3-[4-amino-3-(4-(4-fluorophenoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine Acrylate-2-alkene-1-ketone (compound 34)；

[(R)-3-(4-amino-3-(3-fluoro-4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) nafoxidine-1- Base] ((S)-oxirane-2-base) ketone (compound 35)；

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine-1-base]-2-bromine Acrylate-2-alkene-1-ketone (compound 36)；

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine-1-base]-2-chlorine Acrylate-2-alkene-1-ketone (compound 37)；

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine-1-base]-2-fluorine Acrylate-2-alkene-1-ketone (compound 38)；

(R, Z)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine-1-base]-4-oxygen For but-2-ene cyanogen (compound 39)；

[(R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) nafoxidine-1-base] ((R)-ring Oxidative ethane-2-base) ketone (compound 40)；

(S, E)-1-[3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) nafoxidine-1-base]-3-chlorine Acrylate-2-alkene-1-ketone (compound 41)；

(S, Z)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine-1-base]-3- Bromine acrylate-2-alkene-1-ketone (compound 42)；

(R)-1-[3-[4-amino-3-(4-(pyridine-4-epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine Acrylate-2-alkene-1-ketone (compound 43)；

(S)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine acrylate-2- Alkene-1-ketone (compound 44)；

(R)-1-[3-[4-amino-3-(4-is to toloxyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base]-piperidin-1-yl]-2-bromine Acrylate-2-alkene-1-ketone (compound 45)；

(R)-2-[4-[4-amino-1-[1-(2-bromoacryloyl) piperidines-3-base]-1H-pyrazolo [3,4-d] pyrimidin-3-yl] benzene Epoxide] benzene cyanogen (compound 46)；

(R)-1-[-3-[4-amino-3-(4-(4-4-trifluoromethylphenopendant) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base]-2-bromine acrylate-2-alkene-1-ketone (compound 47)；

(R)-1-[-3-[4-amino-3-(4-(4-vinyl benzene epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base]-2-bromine acrylate-2-alkene-1-ketone (compound 48)；

(R)-1-[-3-[4-amino-3-(4-(4-acetylenylbenzene epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base]-2-bromine acrylate-2-alkene-1-ketone (compound 49)；

(R)-1-[3-[4-amino-3-(4-(3-methoxyphenoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines -1-base]-2-bromine acrylate-2-alkene-1-ketone (compound 50)；

(R)-1-[3-[4-amino-3-(4-(biphenyl-4-epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base]-2-bromine acrylate-2-alkene-1-ketone (compound 51)；

(R)-1-[3-[4-amino-3-(4-(3,4-difluorobenzene epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base]-2-bromine acrylate-2-alkene-1-ketone (compound 52)；

(R)-1-[3-[4-amino-3-(4-(2,3-difluorobenzene epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base]-2-bromine acrylate-2-alkene-1-ketone (compound 53)；

(R)-1-[3-[4-amino-3-(4-(2,4 difluorobenzene epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base]-2-bromine acrylate-2-alkene-1-ketone (compound 54)；

Its concrete structure is as follows:

In the present invention, described term " alkyl " is to include having appointment carbon number purpose side chain or the saturated fat of straight chain Fat race alkyl；As at " C₁-C₁₀Alkyl " defined in for be included in straight chain or branched structure have 1,2,3,4,5, 6, the group of 7,8,9 or 10 carbon atoms.Such as, " C₁-C₁₀Alkyl " specifically include methyl, ethyl, n-pro-pyl, Isopropyl, normal-butyl, the tert-butyl group, isobutyl group, amyl group, hexyl, heptyl, octyl group, nonyl and decyl etc..

In the present invention, described term " alkoxyl " represent alkyl be connected with oxygen atom after generation group, i.e. " RO-", R is alkyl.

In the present invention, described term " alkylthio group " represent alkyl be connected with sulphur atom after generation group, i.e. " RS-", R is alkyl.

In the present invention, described term " alkylamino " expression " NH₃A hydrogen in " replaced by alkyl after amino.

In the present invention, described term " alkylidene " (when including being used alone and being included in other group) means bag Include the side chain of 1～20 carbon atom and the sub-saturated aliphatic hydrocarbyl of straight chain, preferably 1～10 carbon atom, more preferably 1～8 Individual carbon atom, such as methylene, ethylidene, propylidene, isopropylidene, sub-normal-butyl, the sub-tert-butyl group, isobutylidene, Pentylidene, hexylidene, heptamethylene, octylene, nonamethylene, decylene, Asia (4,4-dimethyl amyl group), sub-(2,2,4- Tri-methyl-amyl), alkylene undecyl, sub-dodecyl, and their various isomers.

In the present invention, described term " cycloalkyl " refers to that full carbon monocycle or polycyclic moiety, preferably 3～20 carbon are formed The cycloalkyl of 1～3 ring, more preferably 3～10 carbon, such as: cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, Suberyl, ring octyl group, cyclodecane or cyclo-dodecyl.

In the present invention, when described term " Heterocyclylalkyl " uses individually or as a part for another group at this, Refer to 4～12 yuan of monocycles or the polycyclic moiety comprising 1～4 hetero atom (such as one or more in nitrogen, oxygen and sulfur), its In each ring can contain one or more double bonds, but neither one ring has the pi-electron system of total conjugated.Additionally, Any heterocycloalkyl ring can condense on cycloalkyl, aryl, heteroaryl or heterocycloalkyl ring.Within the range defined herein Heterocyclylalkyl includes but not limited to: oxazoline, oxygen cyclobutyl, pyranose, THP trtrahydropyranyl, azetidinyl, 1,4- Dialkyl group, hexahydro azatropylidene base, piperazinyl, piperidyl, pyrrolidinyl, morpholinyl, thio-morpholinyl, dihydro furan Mutter base, glyoxalidine base, indolinyl, dihydro isoxazolyl, dihydro isothiazolyl, dihydrooxadiazole base, dihydro Oxazolyl, dihydro pyrazinyl, pyrazoline base, dihydropyridine base, dihydro-pyrimidin base, pyrrolin base, dihydro tetrazolium Base, thiodiazoline base, dihydro-thiazolyl, dihydro-thiophene base, dihydro triazolyl, dihydro azetidinyl, tetrahydrochysene Furyl and tetrahydro-thienyl and N-oxide thereof.Heterocyclylalkyl can be through carbon atom therein or hetero atom and other bases Group is attached.

In the present invention, described term " thiazolinyl " refers to containing specifying number the straight of carbon atom and at least one carbon-carbon double bond Chain, side chain or ring-type non-aromatic alkyl.Preferably exist a carbon-carbon double bond, and can exist up to four non-aromatic Carbon-carbon double bond.Thus, " C₂～C₁₂Thiazolinyl " refer to the thiazolinyl with 2～12 carbon atoms.“C₂～C₄Thiazolinyl " refer to tool There is the thiazolinyl of 2～4 carbon atoms, including vinyl, acrylic, 2-methyl-propenyl, 1-butylene base and crotyl. The straight chain of thiazolinyl, side chain or loop section can contain double bond.

In the present invention, described term " alkynyl " refers to containing specifying number the straight of carbon atom and at least one triple carbon-carbon bonds Chain, side chain or cyclic hydrocarbon group.Wherein can there are up to three triple carbon-carbon bonds.Thus, " C₂～C₁₂Alkynyl " refer to tool There is the alkynyl of 2～12 carbon atoms.“C₂～C₄Alkynyl " refer to the alkynyl with 2～4 carbon atoms, including acetenyl, Propinyl, ethyl acetylene base and 2-butyne base etc..

In the present invention, described term " aryl " refer to any stable monocycle that may be up to 7 atoms in each ring or Person's bicyclic carbocyclic, at least one of which ring is aromatic rings；The example of above-mentioned aryl unit includes phenyl, naphthyl, naphthane Base, indanyl, xenyl, phenanthryl, anthryl or acenaphthenyl (acenaphthyl).It is appreciated that at aryl Substituent group is two ring substituents, and one of them ring is in the case of non-aromatic ring, and connection is carried out by aromatic ring.

In the present invention, described term " heteroaryl " represents stable monocycle or the bicyclo-that may be up to 7 atoms in each ring, At least one of which ring is aromatic rings and contains 1-4 the hetero atom selected from O, N and S；Within the range defined herein Heterocyclic aryl includes but not limited to: acridinyl, carbazyl, cinnolines base, quinoxalinyl, pyrazolyl, indyl, benzo Triazolyl, furyl, thienyl, benzothienyl, benzofuranyl, quinolyl, isoquinolyl, oxazolyl, different Oxazolyl, indyl, pyrazinyl, pyridazinyl, pyridine radicals, pyrimidine radicals, pyrrole radicals, tetrahydroquinoline.As following miscellaneous As the definition of ring, " heterocyclic aryl " it should also be understood that the N-oxide derivative being to include any nitrogen-containing hetero aryl.At it Middle heterocyclic aryl substituent group be two ring substituents and ring be non-aromatic ring or do not comprise heteroatomic in the case of, can To understand, connect and by aromatic ring or carry out by comprising the hetero atom of ring respectively.

In the present invention, the described " C determining carbon number range_x1-_y1" alkyl (x1 and y1 is integer), cycloalkyl, miscellaneous Cycloalkyl, aryl and heteroaryl, such as " C_1-12Alkyl ", all represent the C not comprising substituent group_1-12Alkyl.

In the present invention, described term " halogen " represents fluorine, chlorine, bromine, iodine or astatine.

In the present invention, described term " cyano group " represents

For each variable, the combination in any of above-mentioned group is also among considering herein.It is to be understood that carried herein Substituent group and substitute mode on the compound of confession can be selected by those of ordinary skill in the art, in order to provide chemistry Upper stable and that techniques known in the art and technology set forth herein can be used to synthesize compound.

Use Standard synthetic techniques well known by persons skilled in the art or use synthetic method known in the art with described herein Synthetic method combination, the compound of any one in Pyrazolopyrimidine derivative shown in formula I can be synthesized.It addition, Solvent given herein, temperature and other reaction condition can change according to art technology.As further instructing, Following synthetic method can also be utilized.

In some embodiments, it provided herein that the preparation method of Pyrazolopyrimidine derivative shown in formula I and Using method.In some embodiments, compound described herein can use the scheme of following synthesis to synthesize, it is possible to To use and following similar method, by selecting suitable initiation material to carry out synthesising target compound.

Can be synthesized for synthesizing the initiation material of compound described herein or can obtain from commercial source.Retouch herein The compound stated, and other relevant compound with different substituents can use technology well known by persons skilled in the art And Material synthesis, such as at " Encyclopedia of Reagents for Organic Synthesis " (Wiley of Paquette 2009), " Advanced Organic Chemistry " fourth edition of March, (Wiley1992)；Carey and Sundberg " Advanced Organic Chemistry " fourth edition, A volume and B volume (Plenum 2000,2001)；Green With " Protective Groups in Organic Synthesis " third edition of Wuts, (Wiley1999)；Fieser and Fieser " Reagents for Organic Synthesis " the 1-17 volume (John Wiley and Sons, 1991)；《Organic Reactions " the 1-40 volume (John Wiley and Sons, 1991), and " the Comprehensive Organic of Larock Transformations " (VCH PublishersInc., 1999) (being fully incorporated herein by quoting).Preparation is originally The conventional method of the disclosed compound of literary composition can come from reaction known in the art, and this reaction can be by by this area Reagent and condition that technical staff is thought fit are revised, with synthesising target compound molecule.Following synthetic method is permissible Based on instruct utilize.

After each reaction terminates, product can use routine techniques to carry out post processing (separate and purification), including but not Be limited to filter, distill, crystallize, the method such as chromatograph.These products can use conventional method of analysis to characterize, including Physical constant and spectrum data (m.p, HPLC, LC-MS, NMR and optical rotation etc.).Compound described herein can To use synthetic method described herein to be prepared as individual isomer.

Present invention also offers the preparation method of described Pyrazolopyrimidine derivative shown in formula I, it uses following step Rapid: compound II to be carried out coupling reaction with acid ZOH, obtains Pyrazolopyrimidine derivative shown in formula I；

Wherein, the definition of Ar, L, Y, Z and V is the most same as above；Described coupling reaction can use should in this area The conventional method of class reaction and condition.

In the present invention, the preparation method of described Pyrazolopyrimidine derivative shown in formula I, it farther includes following Step: compound III is carried out sloughing the reaction of protection group with acid, obtains described compound II；

Wherein, the definition of Ar, L, Y and V is the most same as above；PG is blocking group, can be Boc (tertiary butyloxycarbonyl Base), CBz (benzyloxycarbonyl group) or Troc (trichloro-ethoxycarbonyl), preferably Boc (tertbutyloxycarbonyl).Described sloughs The reaction of protection group can use conventional method and the condition of such reaction in this area.

In the present invention, the preparation method of described Pyrazolopyrimidine derivative shown in formula I, it farther includes following Step: under palladium chtalyst, carries out Suzuki coupling reaction by compound IV and compound V, obtains described compound III；

Wherein, the definition of Ar, L, Y, V and PG is the most same as above；R ' and R " the most independent for H or alkyl (example Such as C₁～C₄Alkyl, described " C₁～C₄Alkyl " such as methyl, ethyl, propyl group, isopropyl, butyl, different Butyl or the tert-butyl group；Or R ', R " and oxygen atom, boron atom between them be collectively forming circulus (such as)。

Described Suzuki coupling can use conventional method and the condition of such reaction in this area.

In the present invention, the preparation method of described Pyrazolopyrimidine derivative shown in formula I, it farther includes following Step: compound VI and compound V is carried out Mitsunobu reaction, obtains described compound IV；

Wherein, Y is defined as above described；Described Mitsunobu reaction can use the normal of such reaction in this area Rule method and condition.

In the present invention, the preparation method of described Pyrazolopyrimidine derivative shown in formula I, such as method 1 or method 2,

Method 1 comprises the following steps: 3-iodo-1H-pyrazolo [3,4-d] pyrimidine-4-amine (compound VI-1) passes through Mitsunobu Reaction respectively with (R) or (S)-N-Boc-3-hydroxy piperidine and (R) or (S)-N-Boc-3-hydroxypyrrole coupling, Obtain the midbody compound IV-1 of Boc protection.Then, suitable substituted phenylboric acid carries out porpezite in the basic conditions Belong to catalysed cross coupling to react, generate intermediate compound III-1.With acid deprotect after again with Z acid coupling, thus Complete synthesis and obtain target compound I-1.

Method 2 comprises the following steps: 3-iodo-1H-pyrazolo [3,4-d] pyrimidine-4-amine (compound VI-1) passes through Mitsunobu Reaction and (R) or (S)-N-Boc-dried meat ammonia alcohol coupling, obtain the intermediate compound IV-2 of Boc protection.Then, suitably take The phenylboric acid in generation carries out palladium metal catalysed cross coupling in the basic conditions and reacts, and generates intermediate III-2.With acid After deprotection again with Z acid coupling, thus complete synthesis and obtain target compound I-2.

Use synthetic method described herein, and those methods known in the art, obtain this with good yield and purity Pyrazolopyrimidine derivative shown in formula I disclosed in literary composition.The compound prepared according to method disclosed herein passes through ability Conventional method purification known to territory, such as filtration, recrystallization, chromatograph, distillation and combinations thereof.

Present invention also offers described arbitrary compound shown in formula I or its pharmaceutically acceptable salt or its solid is different Structure body, solvate, active metabolite and the prodrug purposes in preparing medicine, can be individually dosed, or With other therapeutic agent medication.Described medicine be used for treating and/or prevent mammal (including people) with suppression cloth Shandong Dun Shi tyrosine kinase (Btk) is active or benefits from, for treatment, the disease that bruton's tyrosine kinase (Btk) activity suppresses Disease, disease or condition of illness.Described medicine can be individually dosed, or with other therapeutic agent medication.Drug combination bag Include but be not limited to amycin, dactinomycin, bleomycin, vinblastine, cisplatin, acivicin；Aclarubicin；Hydrochloric acid Acodazole；Acronine；Adozelesin；Aldesleukin；Altretamine；Ambomycin；Acetic acid ametantrone； Aminoglutethimide；Amsacrine；Anastrozole；Antramycin；Asparaginase；Asperlin；Azacitidine；A Zha replaces Group；Azotomycin；Batimastat；Benzodepa；Bicalutamide；Bisantrene hydrochloride；Two methanesulfonic acid bisnafides；Ratio Folding comes new；Bleomycin Sulphate；Brequinar sodium；Bropirimine；Busulfan；Actinomycin C；Calusterone；Card vinegar Amine；Carbetimer；Carboplatin；Carmustine；Carubicin hydrochloride；Carzelesin；Cedefingol；Chlorambucil； Cirolemycin；Cladribine；Methanesulfonic acid crisnatol；Cyclophosphamide；Cytosine arabinoside；Dacarbazine；Hydrochloric acid is soft red mould Element；Decitabine；Dexormaplatin；Dezaguanine；Dezaguanine mesilate；Diaziquone；Doxorubicin；How soft hydrochloric acid is Compare star；Droloxifene；Droloxifene citrate；Propanoic acid his androsterone bent；Duazomycin；Edatrexate；Hydrochloric acid is according to fluorine bird Propylhomoserin；Elsamitrucin；Enloplatin；Enpromate；Epipropidine；Erbulozole；Esorubicin hydrochloride；Estramustine； Estramustine phosphate sodium；Etanidazole；Etoposide；Etoposide phosphate；Etoprine；CGS-16949A；Method Bundle draws shore；Fenretinide；Floxuridine；Fludarabine phosphate；Fluorouracil；Flurocitabine；Fosquidone；Fostriecin Sodium；Gemcitabine；Gemcitabine hydrochloride；Hydroxyurea；Idarubicin hydrochloride；Ifosfamide；Ilmofosine；White thin Born of the same parents' interleukin II (including recombinant interleukin II or rIL2), Intederon Alpha-2a；Interferon Alpha-2b；Interferon alfa-n1； Alferon N；Interferon beta-1a；Gamma interferon 1-b；Iproplatin；Irinotecan hydrochloride；Lanreotide acetate；Come Bent azoles；Leuprorelin acetate；Liarozole hydrochloride；Lometrexol sodium；Lomustine；Losoxantrone hydrochloride；Masoprocol； Maytansine；Mustine hydrochlcride；Megestrol acetate；Melengestrol acetate；Melphalan；Menogaril；Mercaptopurine；First ammonia Pterin；Methotrexate sodium；Metoprine；Meturedepa；Mitindomide；Mitocarcin；Mitochromine mitocromine B-35251；Mitogillin； Mitomalcin；Mitomycin；Mitosper；Mitotane；Mitoxantrone hydrochloride；Mycophenolic Acid；Nocodazole；Nola Mycin；Ormaplatin；Oxisuran；Pegaspargase；Peliomycin；Pentamustine；Peplomycin sulfate；Perfosfamide； Pipobroman；Piposulfan；Hydrochloric acid piroxantrone；Plicamycin；Plomestane；Porfimer sodium；Porphyromycin；Sprinkle Nimustine；Procarbazine hydrochloride；Puromycin；Puromycin hydrochloride；Pirazofurin；Riboprine；Rogletimide； Safingol；Hydrochloric acid Safingol；Semustine；Simtrazene；Sparfosate sodium；Sparsomycin；Spirogermanium hydrochloride； Spiromustine；Streptonigrin；Streptozocin；Sulofenur；His sharp mould rope；Tecogalan sodium；Ftorafur；Hydrochloric acid replaces Lip river Anthraquinone；Temoporfin；Teniposide；Teroxirone；Testolactone；ITG, thioguanine, phosphinothioylidynetrisaziridine；Thiazole Furan woods；Tirapazamine；Toremifene Citrate；Trestolone acetate, trestolone acetate；Phosphoric acid triciribine；Front three Qu Sha；Glucuronic acid trimetrexate；Triptorelin；Tubulozole hydrochloride；Uracil mustard；Uredepa；Dai Pu Peptide；Verteporfin；Vinblastine sulfate；Vincristine sulfate；Vindesine；Vindesine sulfate；Sulphuric acid vinepidine； Sulphuric acid vinglycinate；Sulphuric acid vinleucinol；Vinorelbine；Sulphuric acid vinrosidine；Sulphuric acid vinzolidine；Vorozole；Folding Buddhist nun's platinum；Zinostatin；Zorubicin hydrochloride, rice torr anthrone, paclitaxel, procarbazine, match TEPA, angiogenesis press down Preparation, camptothecine, dexamethasone, aspirin, acetyl aminophenol, indomethacin, ibuprofen, ketoprofen, Mei Luo Former times health, corticosteroid and adrenocortical steroid.

In the present invention, described disease, disease or the condition of illness bag of benefiting from bruton's tyrosine kinase (Btk) activity suppression Include but be not limited to cancer, diseases associated with inflammation or morbid state, immunity class disease or morbid state, hyperplasia class disease or Morbid state and degeneration class disease or morbid state.

In the present invention, described diseases associated with inflammation or morbid state, immunity class disease or morbid state include but not limited to roar Asthma, inflammatory bowel (including clone disease and ulcerative colitis), appendicitis, blepharitis, bronchiolitis, Tracheitis, bursitis, cervicitis, cholangitis, cholecystitis, colitis, conjunctivitis, cystitis, dacryoadenitis, dermatitis, Dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fascitis, Fibrositis, gastritis, gastroenteritis, hepatitis, hidradenitis suppurativa, laryngitis, mastitis, meningitis, myelitis, Myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, pancreatitis, parotitis, pericarditis, peritoneum Inflammation, pharyngitis, pleuritis, phlebitis, pneumonia (pneumonitis or pneumonia), proctitis, prostatitis, kidney Nephropyelitis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendinitis, tonsillitis, uveitis, vaginitis, Vasculitis, vulvitis, HIV, adult respiratory distress syndrome, bone absorpting disease, chronic obstructive pulmonary disease, chronic pneumonia, Inflammatory dermatosis, atopic dermatitis, Cystic fibrosis, septic shock, sepsis, endotoxin shock, blood Hydraulic power shock, sepsis syndrome, post ischemia reperfusion damage, psoriasis, fibrotic conditions, malignant disease Matter, the transplant rejection of graft versus host disease, autoimmune disease, heteroimmune condition of illness or disease, radiation damage, heavy breathing Breathe heavily, syndrome is infected in oxygen-enriched property injury of lung, microorganism infection and microorganism.

Described autoimmune disease, include but not limited to rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Chauffard-Still disease, juvenile arthritis, lupus, diabetes, myasthenia gravis, struma lymphomatosa, Ao Deshi first shape Adenitis, Graves disease, sjogren syndrome, multiple sclerosis, Guillain Barre syndrome, acute disseminated marrowbrain Inflammation, Addison's disease, heterphoria spasm syndrome, ankylosing spondylitis, antiphospholipid antibody syndrome, aregeneratory Property anemia, autoimmune hepatitis, celiac disease, Goodpasture's syndrome, idiopathic thrombocytopenic purpura, Optic neuritis, scleroderma, primary biliary cirrhosis, conjunctivo-urethro-synovial syndrome, aortic arch syndrome, temporal arteritis, Warm autoimmune hemolytic anemia, Wei Genashi granulomatosis, psoriasis, whole body hair depigmentation, Bei Qiete Family name's disease, confirmed fatigue, autonomic nerve function exception, endometriosis, interstitial cystitis, neuromyotonia, Scleroderma and vulvodynia.

Described heteroimmune condition of illness or disease, include but not limited to graft versus host disease, transplant, transfuse blood, allergy Reaction, anaphylaxis are (such as, to plant pollen, latex, medicine, food, insect toxins, animal hair, animal skins Bits, dust mite or Blatta seu periplaneta calyx allergy), I type allergy, anaphylaxis conjunctivitis, allergic rhinitis and atopic dermatitis.

In the present invention, described cancer includes but not limited to head cancer, thyroid carcinoma, neck cancer, cancer eye, skin carcinoma, mouth Chamber cancer, laryngocarcinoma, esophageal carcinoma, breast cancer, osteocarcinoma, leukemia, bone marrow cancer, pulmonary carcinoma, colon cancer, carcinomaofsigmoid, straight Intestinal cancer, gastric cancer, carcinoma of prostate, breast carcinoma, ovarian cancer, renal carcinoma, hepatocarcinoma, cancer of pancreas, the brain cancer, intestinal cancer, heart cancer, Adrenal carcinoma, subcutaneous tissue cancer, lymph node cancer, melanoma, glioblastoma, B cell proliferative disease.Institute The B cell proliferative disease stated such as diffusivity large B cell lymphoid tumor, follicular lymphoma, chronic lymphocytic drench Bar tumor, chronic lymphocytic leukemia, B cell PL, lymphoplasmacytic lymphoma/Walden B is thin for Si Telun macroglobulinemia, splenic marginal zone lymphoma, plasma cell myeloma, plasmocytoma, knot outer edge area Born of the same parents' lymphoma, lymphoma nodal marginal zone B cell, lymphoma mantle cell, mediastinum (thymus) large B cell lymphoid tumor, Intravascular large B cell lymphoma, lymphoma primary effusion, Burkitt lymphoma/leukemia or lymphoma sample granulation Swollen disease.

Present invention also offers a kind of pharmaceutical composition, it comprises reactive compound and optionally, one or more pharmacy Upper acceptable carrier or excipient；Described reactive compound be described Pyrazolopyrimidine derivative shown in formula I, One in its pharmaceutically acceptable salt, its stereoisomer, solvate, active metabolite and prodrug or Multiple.

Described pharmaceutical composition can use one or more pharmaceutically acceptable carriers to prepare in a conventional manner, institute State carrier and include excipient and adjuvant, its be conducive to being processed as reactive compound can be medicinal preparation.Suitable preparation Depend on the route of administration selected.Technology, carrier and excipient known to any can suitably be used, and such as ability Territory is understood.

Heretofore described pharmaceutical composition refers to the compound of any one and other chemistry in compound described herein example Component such as carrier, stabilizer, antioxidant, disintegrating agent, diluent, dispersant, filler, correctives, suspension The mixture that agent, fluidizer, solubilizing agent, surfactant, wetting agent, thickening agent and/or excipient are formed.Described Pharmaceutical composition beneficially reactive compound gives organism.In putting into practice treatment provided herein or using method, with medicine The reactive compound described herein of therapeutically effective amount is suffered from disease, obstacle or disease to be treated by compositions Mammal.Preferably, described mammal is people.Therapeutically effective amount can change very big, depends on the serious of disease Degree, the age of curee and relative health, the other factors such as effect of compound of use.Described activity Compound can be used alone or is used in combination with one or more curatives of the component as mixture.

Pharmaceutical composition by include at least one Pyrazolopyrimidine derivative shown in formula I described herein any one Compound, using the form of free acid or free alkali as active component, or becomes using pharmaceutically acceptable salt form as activity Point.It addition, method described herein and pharmaceutical composition include use N-oxide, crystal form (also referred to as polymorphic), And these have the active metabolite of Pyrazolopyrimidine derivative shown in formula I of same type.In some cases, Pyrazolopyrimidine derivative shown in formula I can exist with tautomer.All of tautomer is included in and carries herein In the range of the compound of confession.It addition, Pyrazolopyrimidine derivative shown in formula I described herein can be with non-solvated The form of thing and solvate exists together with pharmaceutically acceptable solvent such as water, ethanol etc..Provided herein such as Formulas I The solvate forms of shown Pyrazolopyrimidine derivative is recognized as being disclosed.

The pharmaceutical composition that the present invention describes can give curee by multiple route of administration, and described route of administration includes But be not limited to be administered orally, parenteral (such as intravenous, subcutaneous, intramuscular), intranasal, buccal, locally, rectum or warp Skin route of administration.The dosage form of the pharmaceutical composition that the present invention describes includes but not limited to that aqueous liquid dispersion, self emulsifying disperse Body, solid solution, liposomal dispersion agent, aerosol, solid dosage forms, powder, immediate release formulations, controlled release preparation, instant system Agent, tablet, capsule, pill, slow releasing preparation, delayed release dosage system, pulsation delivery formulations, many granular preparations and mixing Instant and controlled release preparation.

Pharmaceutical preparations for oral can be by by one or more solid excipients and one or more described hereinization Compound mixes and obtains, and optionally grinds the mixture obtained, if it is desired, mix after adding suitable adjuvant Thing is pelletized, to obtain tablet or lozenge core.Suitably excipient includes such as filler such as saccharide, including lactose, sucrose, Mannitol or sorbitol；Cellulose preparation, such as corn starch, wheaten starch, rice starch, potato starch, gelatin, Tragakanta, methylcellulose, microcrystalline Cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose；Etc., example Such as polyvinylpyrrolidone (PVP or polyvidone) or calcium phosphate.It is possible if desired to addition disintegrating agent, such as, cross-link friendship Connection carmethose, polyvinylpyrrolidone, agar or alginic acid or its salt such as sodium alginate.

The pharmaceutical preparation that can orally use includes the push-in type capsule by Gelatin production and by gelatin and plasticizer such as The soft seal capsule that glycerol or sorbitol manufacture.Described push-in type capsule can comprise active component, and it is mixed in filler such as Lactose, binding agent such as starch and/or lubricant such as Pulvis Talci or magnesium stearate, and optional stabilizer.In soft capsule, Reactive compound can be dissolved or suspended in suitable liquid, such as fatty oil, liquid Paraffin or liquid macrogol. Furthermore it is possible to addition stabilizer.All formulations for oral administration should be the dosage form being suitable for this administration.

In some embodiments, solid dosage forms disclosed herein can in form of tablets (include suspendible tablet, dissolving tablet, Chew disintegrating tablet, fast disintegrating tablet, effervescent tablet or Caplet), pill, powder (include the powder of aseptic packaging, Nonessential powder or effervescent powder), capsule (includes soft capsule or hard capsule, such as by the gelatin system of animal origin The capsule made or the capsule manufactured with the HPMC of plant origin or " decentralized capsule "), solid dispersion, solid solution, Biological erodable dosage form, control delivery formulations, pulsatile release dosage forms, many bead dosage form, piller, granule or aerosol Agent.In other embodiments, described pharmaceutical preparation is powder type.In other embodiment, described medicine system Agent is tablet form, includes but not limited to dissolving tablet.It addition, pharmaceutical preparation described herein can be as single glue Wafer or with the medication of many capsule formulations.In some embodiments, pharmaceutical preparation is with two or three or four capsules Agent or tablet medication.

Pharmaceutical composition will include at least one Pyrazolopyrimidine derivative shown in formula I described herein any one The preparation of compound, is suitable for intramuscular, subcutaneous or intravenous injection, can include physiologically acceptable sterilized water or non-aqueous Solution, dispersion, suspensoid or Emulsion and sterile powder injection, described sterile powder injection is for again preparing aseptic note Penetrate solution or dispersion.The suitably example of aqueous and non-aqueous carrier, diluent, solvent or solvent includes water, second Alcohol, polyhydric alcohol (propylene glycol, Polyethylene Glycol, glycerol, Ke Liemo (cremophor) etc.), its suitable mixture, plant Oil (such as olive oil) and injection organic ester such as ethyl oleate.Suitable mobility can be maintained, such as by using coating Such as lecithin；During dispersion, by the granular size needed for maintaining；With by use surfactant.It is suitable for skin The preparation of hemostasis can also comprise additive such as preservative, wetting agent, emulsifying agent and dispersant.Can be by various anti- Bacteriocin and antifungal guarantee to prevent growth of microorganism, such as p-Hydroxybenzoate, chlorobutanol, phenol, mountain Pears acid, etc..Can also as required, including isotonic agent, such as sugar, sodium chloride etc..Extend injectable drug form Absorption can by use postpone absorbent bring, such as aluminum monostearate and gelatin.

The application further relates in pharmaceutical field known for including the various medicines of the conveying of ophthalmic, intranasal and in ear Compositions.Pharmaceutical formulation includes the aqueous ophthalmic solution of described reactive compound, and it can such as eye drop water-soluble Property form exist, or with gellan gum or hydrogel；Ophthalmic ointment；Ophthalmic suspension, such as microgranule, be suspended in liquid and carry The little polymer particles comprising medicine in body medium, fat-soluble preparation, and microsphere；And ocular inserts.In order to stable Property and comfortableness, these suitable pharmaceutical preparatioies most frequently and are preferably manufactured as aseptic, the isotonic and preparation of buffering. Pharmaceutical composition also includes drop and spray, and it is simulated nasal discharge the most in many aspects and guarantees normal ciliary effect Maintain.As known to those skilled in the art, suitable preparation, the most often with the most isotonic, keeps pH The slight buffering of value 5.5 to 6.5, and most frequently and preferably include antimicrobial preservative and suitable drug stabilizing agent. Pharmaceutical preparation in ear transhipment includes suspension and the ointment in topical application.General for these aural preparationses Logical solvent includes G & W.Can be through oral, parenteral (intravenous, intramuscular, subcutaneous etc.), through pulmonary, office The administering mode such as portion, skin is applied to mammal, such as people.People's dosage of the compounds of this invention can be about 0.1mg To about 1000mg scope.

Without prejudice to the field on the basis of common sense, above-mentioned each optimum condition, can combination in any, obtain the present invention each preferably Example.

Agents useful for same of the present invention and raw material are the most commercially.

In the present invention, room temperature refers to ambient temperature, is 10 DEG C～25 DEG C.

The most progressive effect of the present invention is: the Pyrazolopyrimidine derivative shown in formula I of the present invention is to Bu Ludun cheese Histidine kinase has good inhibitory action, and the growth to tumor cell has good vivo and vitro inhibitory activity, has good Good market-oriented prospect.

Detailed description of the invention

Embodiment and biological example is prepared so that this invention is expanded on further, these embodiments master below by particular compound It is used for specifically describing in more detail being used, and is not to be construed as limiting in any form the present invention.Following enforcement The experimental technique of unreceipted actual conditions in example, conventionally and condition, or selects according to catalogue.

1. compound prepares part:

Embodiment 1:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-2-bromine acrylate-2-alkene-1-ketone (compound 1)

Compound 1

A. the synthesis of (R)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidines

By 4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine (10g, 38mmol), (S)-1-Boc-3-hydroxy piperidine (17g, 85mmol), triphenylphosphine (20g, 76mmol) adds in there-necked flask, adds THF (120ml), is cooled to 0 DEG C, Dropping diisopropyl azodiformate (DIAD) (15.2g, 76mmol) and oxolane (THF) (30ml) mix Closing liquid, about 1h dropping is complete, is slowly increased to room temperature reaction overnight.Being spin-dried for by reactant liquor, add water, ethyl acetate extracts, It is dried, concentrates rear pillar chromatography purification and obtain product (R)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) Piperidines (13.5g, yield 80%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.38 (s, 1H), 6.02 (bs, 2H), 4.82-4.64(m,1H),4.31-4.02(m,2H),3.44-3.20(m,1H),2.95-2.65(m,1H),2.25-2.08(m,2H), 1.95-1.58(m,2H),1.42(s,9H).

B.4-the synthesis of phenoxyphenyl boronic acid

4-bromo biphenyl ether (100g, 0.402mol) is dissolved in anhydrous THF (800mL), under nitrogen protection, is cooled to-78 DEG C, dropping n-BuLi (n-BuLi) (177mL, 0.442mol), temperature keeps less than-65 DEG C, after stirring 1h, Continue to keep temperature-60 DEG C, dropping triisopropyl borate ester (90g, 0.482mol), be slowly increased to 0 DEG C, stir 3h. Add water (300mL) under ice bath to be stirred overnight.Reactant liquor concentrates removes organic facies, is adjusted to 12N concentrated hydrochloric acid PH 1～2, separate out solid, filtration drying obtains product (77g, productivity 90%).¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.0 (s, 2H), 7.80 (d, J=8.4Hz, 2H), 7.41 (t, J=7.6Hz, 2H), 7.16 (t, J=7.2Hz, 1H), 7.03 (d, J=8.4Hz, 2H), 6.94 (d, J=8.4Hz, 2H).

C. the synthesis of (R)-1-Boc-3-(4-amino-3-(4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidines

(R)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidines (12.8 is added in there-necked flask G, 29mmol), 4-phenoxyphenyl boronic acid (6.8g, 32mmol), PdCl₂(dppf) (0.5g, 0.69mmol), Sodium carbonate (6.1g, 58mmol), Isosorbide-5-Nitrae-dioxane (160ml) and water (40ml), after nitrogen displacement, be warming up to 80 DEG C of reactions are overnight.After some plate confirms that reaction completes, filtering, be spin-dried for, add water, ethyl acetate extracts, and is dried, and concentrates Rear pillar chromatography purification obtains product (8.5g, productivity 60%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.38 (s, 1H), 7.64 (d, J=8.4Hz, 2H), 7.42-7.38 (m, 2H), 7.24-7.04 (m, 5H), 6.18 (br, 2H),4.86-4.81(m,1H),4.18-4.04(m,1H),3.72-3.68(m,1H),3.18-3.01(m, 1H),2.84-2.78(m,1H),2.36-2.18(m,1.5H),2.08-1.84(m,1.5H),1.78-1.68(m, 1H),1.44(s,9H)。

D. the synthesis of (R)-3-(4-phenoxy phenyl)-1-(piperidines-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia

In reaction bulb, (R)-1-Boc-3-(4-amino-3-(4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperazine Pyridine (8g, 16.4mmol) joins in trifluoroacetic acid (40ml) and dichloromethane (40ml) mixed solution, and room temperature is anti- Should be overnight.After some plate (TLC) confirms that reaction completes, use saturated Na₂CO₃Solution is neutralized to after pH is 7-8, with two Chloromethanes (DCM) extract, organic facies merge, anhydrous sodium sulfate is dried, concentrate rear pillar chromatography purification obtain product (5.4g, Yield 85%).¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.25 (s, 1H), 7.67 (d, J=8.4Hz, 2H), 7.44 (t, J=8.4Hz, 2H), 7.17～7.30 (m, 5H), 4.78 (m, 1H), 3.31～3.40 (m, 1H), 3.22～3.30 (m, 1H), 3.17～3.21 (m, 1H), 3.00～3.10 (m, 1H), 2.60～2.75 (m, 1H), 1.99～2.18 (m, 2H), 1.59～1.83 (m,2H)。

(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine third -2-alkene-1-ketone (compound 1)

By (R)-3-(4-phenoxy phenyl)-1-(piperidines-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia (193mg, 0.5mmol), 2-bromopropene acid (88mg, 0.58mmol), I-hydroxybenzotriazole (HOBT) (95mg, 0.7mmol) and 1- Ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (134mg, 0.7mmol), is dissolved in anhydrous DCM (10ml) in, add diisopropylethylamine (DIEA) (271mg, 2.1mmol), under room temperature, stir 8h.Add Washing after diluted ethyl acetate, aqueous phase is extracted with ethyl acetate 2 times again, merges organic facies, and anhydrous sodium sulfate is dried, Evaporating column chromatography purification obtains compound 1 (195mg, productivity 75%).

¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.38 (s, 1H), 7.64 (d, J=8.4Hz, 2H), 7.46-7.38 (m, 2H), 7.22-7.16 (m, 3H), 7.08 (d, J=8.4Hz, 2H), 6.08-6.02 (s, 1H),5.92-5.86(m,1H),4.94-4.98(m,1H),4.66-4.64(m,0.5H),4.48-4.52(m, 0.5H),4.14-4.11(m,0.5H),3.96-3.99(m,0.5H),3.78-3.76(m,0.5H),3.54-3.50 (m,0.5H),3.24-3.21(m,0.5H),2.94-2.90(m,0.5H),2.38-2.34(m,2H), 2.08-2.04(m,1H),1.86-1.82(m,1H)。

Embodiment 2:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-2-chlorine acrylate-2-alkene-1-ketone (compound 2)

With compound 1 prepare similar method, with 2-chloracrylic acid as raw material, compound 2 (72% productivity) can be prepared.¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.28 (s, 1H), 7.62 (d, J=8Hz, 2H), 7.41 (t, J=8Hz, 2H),7.24-7.08(m,5H),5.71-5.64(m,2H),4.98-4.90(m,1H),4.72-4.66(m,0.5 H),4.58-4.52(m,0.5H),4.18-4.12(m,0.5H),3.99-3.95(m,0.5H),3.80-3.78(m,0.5H), 3.57-3.52(m,0.5H),3.33-3.28(m,0.5H),2.96-2.90(m,0.5H),2.42-2.25(m,2H),2.09-2.01 (m,1H),1.85-1.77(m,1H)。

Embodiment 3:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-2-fluorine acrylate-2-alkene-1-ketone (compound 3)

With compound 1 prepare similar method, with 2-perfluoroalkyl acrylate as raw material, compound 3 (68% productivity) can be prepared.¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.28 (s, 1H), 7.62 (d, J=8Hz, 2H), 7.41 (t, J=8Hz, 2H),7.24-7.08(m,5H),5.71-5.64(m,2H),4.98-4.90(m,1H),4.72-4.66(m,0.5H), 4.58-4.52(m,0.5H),4.18-4.12(m,0.5H),3.99-3.95(m,0.5H),3.80-3.78(m,0.5H), 3.57-3.52(m,0.5H),3.33-3.28(m,0.5H),2.96-2.90(m,0.5H),2.42-2.25(m,2H), 2.09-2.01(m,1H),1.85-1.77(m,1H)。

Embodiment 4:(R, E)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-4,4,4-tri-fluoro-but-2-ene-1-ketone (compound 4)

With compound 1 prepare similar method, be raw material with trans-4,4,4-trifluorobutene acid, compound 4 can be prepared (69% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.38 (s, 1H), 7.64 (d, J=8.4Hz, 2H), 7.38-7.34 (m, 2H), 7.21-7.14 (m, 3H), 7.08 (d, J=8.4Hz, 2H), 7.03-6.95 (m,1H),6.75-6.64(m,1H),5.53-5.51(br,2H),4.91-4.78(m,1.5H),4.41-4.36 (m,0.5H),4.14-4.12(m,0.5H),3.96-3.92(m,0.5H),3.74-3.70(m,0.5H), 3.52-3.48(m,0.5H),3.34-3.29(m,0.5H),3.26-3.18(m,0.5H),2.52-2.46(m, 1H),2.32-2.22(m,1H),2.16-2.02(m,2H)。

Embodiment 5:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-2-trifluoromethyl acrylate-2-alkene-1-ketone (compound 5)

With compound 1 prepare similar method, with 2-(trifluoromethyl) acrylic acid as raw material, compound 5 can be prepared (58% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.34 (s, 1H), 7.61 (d, J=8.4Hz, 2H), 7.41-7.39 (m, 2H), 7.20-7.14 (m, 3H), 7.08 (d, J=8.4Hz, 2H), 6.06-6.03 (m,1H),5.73-5.69(m,1H),4.95-4.86(m,1H),4.80-4.61(m,1H),4.12-4.06(m, 0.5H),3.92-3.84(m,0.5H),3.78-3.71(m,0.5H),3.61-3.52(m,0.5H), 3.34-3.26(m,0.5H),2.98-2.88(m,0.5H),2.42-2.26(m,2H),2.08-2.01(m,1H), 1.86-1.72(m,1H)。

Embodiment 6:(R, Z)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-4-oxo but-2-ene cyanogen (compound 6)

A. (R, Z)-4-(3-(and 4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base]-piperidino]-4-oxygen The synthesis of-2-butylene cyanogen

By (R)-3-(4-phenoxy phenyl)-1-(piperidines-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia (193mg, 0.5mmol), Maleamic acid (67mg, 0.58mmol), I-hydroxybenzotriazole (HOBT) (95mg, 0.7mmol) and 1- Ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (134mg, 0.7mmol), is dissolved in anhydrous DCM (10ml) in, add diisopropylethylamine (DIEA) (271mg, 2.1mmol), under room temperature, stir 8h.Add Washing after diluted ethyl acetate, aqueous phase is extracted with ethyl acetate 2 times again, merges organic facies, and anhydrous sodium sulfate is dried, Evaporating column chromatography purification obtains compound 6 (140mg, productivity 60%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.34 (s, 1H), 7.61 (d, J=8.0Hz, 2H), 7.41-7.37 (m, 2H), 7.20-7.14 (m, 3H), 7.08 (d, J=8.0Hz, 2H), 6.99-6.93 (t, 1H, J=11.6Hz), 5.77 (d, J₁=11.6Hz, 0.5H),5.70(d,J₁=11.6Hz, 0.5H), 4.93-4.89 (m, 1.5H), 4.46-4.42 (m, 0.5H), 4.00-3.86(m,1H),3.52-3.51(m,0.5H),3.50-3.48(m,0.5H),3.33-3.31(m, 0.5H),3.18-3.12(m,0.5H),2.48-2.36(m,2H),2.12-2.07(m,1H),1.86-1.78(m, 1H)。ESI-MS(m/z):466.2[M+H]⁺。

Embodiment 7:(R, E)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-4-oxo but-2-ene acid methyl ester (compound 7)

With compound 1 prepare similar method, with monomethyl fumarate as raw material, (66% produces can to prepare compound 7 Rate).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.38 (s, 1H), 7.62 (d, J=8.4Hz, 2H), 7.41-7.24 (m, 3H), 7.21-7.17 (m, 3H), 7.09 (d, J=8.4Hz, 2H), 6.95 (d, J=15.6 Hz, 0.5H), 6.75 (d, J=15.6Hz, 0.5H), 4.92-4.79 (m, 1.5H), 4.43-4.37 (m, 0.5H),4.14-4.10(m,0.5H),4.03-3.95(m,0.5H),3.92-3.88(m,0.5H),3.81(s, 3H),3.49-3.46(m,0.5H),3.27-3.24(m,0.5H),3.11-3.06(m,0.5H),2.42-2.21 (m,2H),2.10-2.02(m,1H),1.81-1.68(m,1H)。

Embodiment 8:(R, E)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-3-chlorine acrylate-2-alkene-1-ketone (compound 8)

With compound 1 prepare similar method, with trans-chlorallylene acid as raw material, compound 8 (67% can be prepared Productivity).¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.42 (s, 1H), 7.63 (d, J=8.4Hz, 2H), 7.42-7.36 (m, 2H), 7.29-7.12 (m, 4H), 7.08 (d, J=8.4Hz, 2H), 6.76-6.66 (m, 1H),5.92-5.88(br,2H),4.87-4.84(m,1.5H),4.48-4.45(m,0.5H),4.12-4.08(m, 0.5H),4.02-3.98(m,0.5H),3.87-3.84(m,0.5H),3.42-3.38(m,0.5H), 3.28-3.12(m,0.5H),3.06-2.98(m,0.5H),2.42-2.33(m,2H),2.12-1.98(m, 2H)。

Embodiment 9:(R, Z)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-3-bromine acrylate-2-alkene-1-ketone (compound 9)

With compound 1 prepare similar method, with cis-3-bromopropene acid as raw material, compound 9 (63% can be prepared Productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.39 (s, 1H), 7.64 (d, J=8.4Hz, 2H), 7.42-7.36 (m, 2H), 7.20-7.16 (m, 3H), 7.06 (d, J=8.4Hz, 2H), 6.84-6.78 (m, 1H), 6.60 (d, J=8.0Hz, 0.5H), 6.53 (d, J=8.0Hz, 0.5H), 5.61-5.59 (br, 2H), 4.95-4.92(m,1.5H),4.89-4.85(m,0.5H),4.18-4.06(m,0.5H),3.96-3.82(m, 1H),3.48-3.42(m,0.5H),3.24-3.18(m,0.5H),2.98-2.94(m,0.5H), 2.42-2.22(m,2H),2.06-1.98(m,2H)。

Embodiment 10:(R, E)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-3-bromine acrylate-2-alkene-1-ketone (compound 10)

With compound 1 prepare similar method, with trans-3-bromopropene acid as raw material, compound 10 (62% can be prepared Productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.40 (s, 1H), 7.64 (d, J=8.4Hz, 2H), 7.58-7.54 (m, 1H), 7.46-7.38 (m, 2H), 7.19-7.14 (m, 3H), 7.08 (d, J=8.4Hz, 2H),7.02-6.90(m,1H),5.52-5.51(br,2H),4.87-4.68(m,1.5H),4.45-4.41(m, 0.5H),4.06-4.02(m,0.5H),3.97-3.93(m,0.5H),3.87-3.84(m,0.5H), 3.46-3.42(m,0.5H),3.32-3.21(m,0.5H),3.18-3.06(m,0.5H),2.44-2.41(m, 1H),2.36-2.26(m,1H),2.03-1.96(m,2H)。

Embodiment 11:(R, E)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-4-oxo but-2-enamides (compound 11)

A. the synthesis of (E)-4-amino-4 oxos-2-butylene acid methyl ester

Monomethyl fumarate (15g, 115mmol) is dissolved in toluene 20ml, adds thionyl chloride (20ml), add Hot reflux 4 hours, concentrates.Under ice bath, by NH₃THF solution (80ml) be added drop-wise in the acyl chlorides of above-mentioned preparation, Drip complete, room temperature reaction 1h.With aqueous citric acid solution washing to pH equal to 6, ethyl acetate extracts, and organic facies is again Use saturated NaHCO₃Washing, is dried, is concentrated to give (E)-4-amino-4 oxos-2-butylene acid methyl ester (8.9g, productivity 60%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 6.95 (d, J=15.2Hz, 1H), 6.84 (d, J=15.6Hz, 1H),5.88-5.84(m,1H),5.83-5.81(m,1H),3.82(s,3H)。

A. the synthesis of (E)-4-amino-4 oxos-2-butylene acid

(E)-4-amino-4 oxos-2-butylene acid methyl ester (4g, 31mmol) is dissolved in methanol (40ml), adds LiOH (1.4 G, 62mmol) and water (2ml), 4h is stirred at room temperature.Concentrate, add 2M HCl regulation pH and be equal to 3, ethyl acetate Extract twice, merge organic facies, be dried, concentrate, obtain (E)-4-amino-4 oxos-2-butylene acid (2.7g, productivity 75%). ESI-MS(m/z):116.1[M+H]⁺。

C. (R, E)-4-(3-(and 4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base]-piperidino]-4-oxygen The synthesis of generation-2-butylene amide

With compound 1 prepare similar method, with (E)-4-amino-4 oxos-2-butylene acid as raw material, chemical combination can be prepared Thing 11 (60% productivity).¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.36 (s, 1H), 7.62 (d, J=8.4 Hz, 2H), 7.48-7.36 (m, 3H), 7.18-7.12 (m, 3H), 7.07 (d, J=8.4Hz, 2H), 7.02-6.88(m,1H),5.83-5.81(br,2H),4.87-4.81(m,1.5H),4.51-4.48(m,0.5H), 4.21-4.19(m,0.5H),4.17-4.15(m,0.5H),3.87-3.84(m,0.5H),3.38-3.34(m, 0.5H),3.26-3.19(m,0.5H),2.98-2.96(m,0.5H),2.42-2.38(m,1H),2.34-2.28 (m,1H),2.06-1.98(m,1H),1.78-1.72(m,1H)。ESI-MS(m/z):484.7[M+H]⁺。

Embodiment 12:(R)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] tetrahydrochysene Pyrroles's-1-base] synthesis of-2-fluorine acrylate-2-alkene-1-ketone (compound 12)

A. the synthesis of (R)-1-Boc-2-((4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) methyl) nafoxidine

4-amino-3-iodo-1H-azoles is coughed up also [3,4-D] pyrimidine (10g, 38mmol), and Boc-D-dried meat ammonia alcohol (17.1g, 85 Mmol), triphenylphosphine (20g, 76mmol) adds in there-necked flask, adds THF (120ml), is cooled to 0 DEG C, Dropping DIAD (15.2g, 76mmol) and THF (30ml) mixed liquor, about 1h dropping is complete, is slowly increased to room Temperature reaction is overnight.Being spin-dried for by reactant liquor, add water, ethyl acetate extracts, and is dried, and concentrates rear pillar chromatography purification and obtains product (R)-1-Boc-2-((4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) methyl) nafoxidine (11g, yield 65%). ESI-MS(m/z):445.2[M+H]⁺。

B. (R)-1-Boc-2-((4-amino-3-(4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) methyl) tetrahydrochysene The synthesis of pyrroles

(R)-1-Boc-2-((4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) methyl) four is added in there-necked flask Hydrogen pyrroles (12.9g, 29mmol), 4-phenoxyphenyl boronic acid (6.8g, 32mmol), Pd (dppf) Cl₂(0.5g, 0.69mmol), sodium carbonate (6.1g, 58mmol), Isosorbide-5-Nitrae-dioxane (160ml) and water (40ml), nitrogen is put After changing, it is warming up to 80 DEG C of reactions overnight.After some plate confirms that reaction completes, filtering, be spin-dried for, add water, ethyl acetate extracts, It is dried, concentrates rear pillar chromatography purification and obtain product (9.2g, productivity 65%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.34 (s, 1H), 7.64 (d, J=8.4Hz, 2H), 7.41-7.37 (m, 2H), 7.17-7.13 (m, 3H), 7.09-7.07(m,2H),4.61-4.34(m,3H),3.40-3.26(m,2H),1.84-1.78(m,4H), 1.42(s,9H)。

C. the synthesis of (R)-3-(4-phenoxy phenyl)-1-(nafoxidine-2-methyl base)-1H-pyrazolo [3,4-D] pyrimidine-4-amino

In reaction bulb, (R)-1-Boc-2-((4-amino-3-(4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) Methyl) nafoxidine (7.98g, 16.4mmol) joins trifluoroacetic acid (40ml) and dichloromethane (40ml) mixes molten In liquid, room temperature reaction is overnight.After some plate confirms that reaction completes, use saturated Na₂CO₃Solution is neutralized to after pH is 7-8, Extracting with DCM, organic facies merges, and anhydrous sodium sulfate is dried, and concentrates rear pillar chromatography purification and obtains product (5.1g, yield 80%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.26 (s, 1H), 7.58 (d, J=8.4Hz, 2H), 7.36(m,2H),7.16(m,1H),7.06(m,4H),4.86-4.77(m,2H),4.32-4.30(m,1H), 3.52-3.46(m,1H),3.36-3.32(m,1H),2.22-2.18(m,1H),2.06-1.96(m,2H), 1.86-1.78(m,1H)。

D. (R)-1-(2-((4-amino-3-(4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) methyl) nafoxidine -1-base) synthesis of-2-fluoropropene-1-ketone

By (R)-3-(4-phenoxy phenyl)-1-(nafoxidine-2-methyl base)-1H-pyrazolo [3,4-D] pyrimidine-4-amino (193mg, 0.5mmol), 2-perfluoroalkyl acrylate (52mg, 0.58mmol), HOBT (95mg, 0.7mmol) and 1-ethyl-(3- Dimethylaminopropyl) carbodiimide hydrochloride (EDCI) (134mg, 0.7mmol), it is dissolved in anhydrous DCM (10ml) In, add DIEA (271mg, 2.1mmol), under room temperature, stir 8h.Wash after adding diluted ethyl acetate, aqueous phase Being extracted with ethyl acetate 2 times again, merge organic facies, anhydrous sodium sulfate is dried, and evaporating column chromatography purification obtains compound 12 (160mg, productivity 70%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.36 (s, 1H), 7.62 (d, J= 8.4Hz, 2H), 7.46-7.42 (m, 2H), 7.28-7.22 (m, 3H), 7.16 (d, J=8.4Hz, 2H), 5.46(dd,J₁=46.4Hz, J₂=3.2Hz, 1H), 5.12 (dd, J₁=15.6Hz, J₂=2.8Hz, 1H), 4.75-4.71(m,3H),3.67-3.65(m,1H),3.58-3.56(m,1H),1.86-1.82(m,2H), 1.78-1.68(m,2H)。

Embodiment 13:(R)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] tetrahydrochysene Pyrroles's-1-base] synthesis of-2-bromine the third 2-alkene-1-ketone (compound 13)

With compound 12 prepare similar method, with the acid of 2-bromopropene as raw material, (58% produces can to prepare compound 13 Rate).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.30 (s, 1H), 7.57 (d, J=8.4Hz, 2H), 7.40-7.38 (m, 2H), 7.20-7.13 (m, 3H), 7.08 (d, J=8.4Hz, 2H), 5.98 (d, J=2 Hz, 1H), 5.81 (d, J=2Hz, 1H), 4.72-4.68 (m, 3H), 3.68-3.62 (m, 1H), 3.52-3.48(m,1H),2.02-1.94(m,2H),1.92-1.86(m,2H)。

Embodiment 14:(R)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] tetrahydrochysene Pyrroles's-1-base] synthesis of-2-chlorine acrylate-2-alkene-1-ketone (compound 14)

With compound 14 prepare similar method, with 2-chloracrylic acid as raw material, (52% produces can to prepare compound 13 Rate).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.31 (s, 1H), 7.61 (d, J=8.4Hz, 2H), 7.39 (t, J= 8Hz, 2H), 7.22-7.08 (m, 5H), 5.68 (d, J=2Hz, 1H), 5.63 (d, J=2Hz, 1H), 4.77-4.66 (m, 3 H),3.58-3.57(m,1H),3.45-3.43(m,1H),2.11-2.04(m,1H),1.92-1.88(m,3H)。

Embodiment 15:(R, E)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] Nafoxidine-1-base] synthesis of-4,4,4-trifluoro but-2-ene-1-ketone (compound 15)

With compound 12 prepare similar method, be raw material with trans-4,4,4-trifluorobutene acid, compound can be prepared 15 (67% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.38 (s, 1H), 7.66 (d, J=8.4Hz, 2H), 7.38-7.32 (m, 2H), 7.19-7.15 (m, 3H), 7.08 (d, J=8.4Hz, 2H), 6.82-6.78 (m,1H),6.69-6.67(m,1H),5.52-5.50(br,2H),4.78-4.67(m,2H),4.64-4.59(m, 1H),3.76-3.69(m,1H),3.56-3.48(m,1H),2.10-2.05(m,2H),1.78-1.66(m, 2H)。

Embodiment 16:(R, E)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] Nafoxidine-1-base] synthesis of-4-oxo but-2-ene acid methyl ester (compound 16)

With compound 12 prepare similar method, with monomethyl fumarate as raw material, compound 16 (70% can be prepared Productivity).¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.28 (s, 1H), 7.60 (d, J=8.4Hz, 2H), 7.40-7.36 (m, 2H), 7.22-7.08 (m, 5H), 6.98 (d, J=15.2Hz, 0.5H), 6.83 (d, J= 15.2Hz, 1H), 6.60 (d, J=15.2Hz, 0.5H), 4.71-4.68 (m, 2H), 4.39-4.36 (m, 1H), 3.79(s,3H),3.72-3.67(m,1H),3.59-3.56(m,1H),2.16-2.04(m,2H), 2.01-1.94(m,2H)。

Embodiment 17:(R, Z)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] Nafoxidine-1-base] synthesis of-4-oxo but-2-ene cyanogen (compound 17)

With compound 12 prepare similar method, with maleamic acid as raw material, (65% produces can to prepare compound 17 Rate).¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.36 (s, 1H), 7.62 (d, J=8.4Hz, 2H), 7.42-7.38 (m, 2H), 7.20-7.16 (m, 5H), 6.95 (d, J=11.6Hz, 0.5H), 6.82 (d, J= 11.2Hz, 0.5H), 5.76 (d, J=11.2Hz, 0.5H), 5.62-5.69 (m, 2.5H), 4.81-4.78 (m, 1H),4.77-4.68(m,1H),4.56-4.52(m,1H),3.78-3.72(m,1H),3.48-3.42(m, 1H),2.02-1.98(m,2H),1.86-1.78(m,2H)。

Embodiment 18:(S)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] tetrahydrochysene Pyrroles's-1-base] synthesis of-2-fluorine acrylate-2-alkene-1-ketone (compound 18)

With compound 12 prepare similar method, with Boc-L-dried meat ammonia alcohol and 2-perfluoroalkyl acrylate as raw material, can preparing Compound 18 (68% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.42 (s, 1H), 7.62 (d, J=8.4 Hz, 2H), 7.44-7.38 (m, 2H), 7.18-7.12 (m, 3H), 7.06 (d, J=8.4Hz, 2H), 5.48 (d, J=48Hz, 1H), 5.14 (d, J=2.4Hz, 1H), 4.75-4.67 (m, 2.5H), 4.52-4.48 (m, 0.5H),3.65-3.60(m,1H),3.50-3.48(m,1H),1.96-1.88(m,2H),1.86-1.68(m, 2H)。

Embodiment 19:(S)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] tetrahydrochysene Pyrroles's-1-base] synthesis of-2-chlorine acrylate-2-alkene-1-ketone (compound 19)

With compound 18 prepare similar method, with 2-chloracrylic acid as raw material, (64% produces can to prepare compound 19 Rate).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.34 (s, 1H), 7.62 (d, J=8.4Hz, 2H), 7.44-7.38 (m, 2H), 7.28-7.20 (m, 3H), 7.10 (d, J=8.4Hz, 2H), 5.68 (d, J=2.4 Hz, 1H), 5.64 (d, J=2.4Hz, 1H), 4.82-4.68 (m, 3H), 3.58-3.52 (m, 1H), 3.48-3.42(m,1H),2.14-2.10(m,1H),1.95-1.78(m,3H)。

Embodiment 20:(S)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] tetrahydrochysene Pyrroles's-1-base] synthesis of-2-bromine acrylate-2 alkene-1-ketone (compound 20)

With compound 18 prepare similar method, with the acid of 2-bromopropene as raw material, (64% produces can to prepare compound 20 Rate).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.29 (s, 1H), 7.59 (d, J=8.4Hz, 2H), 7.44-7.40 (m, 2H), 7.23-7.19 (m, 3H), 7.09 (d, J=8.4Hz, 2H), 6.00 (d, J=2 Hz, 1H), 5.83 (d, J=2Hz, 1H), 4.77-4.64 (m, 3H), 3.58-3.56 (m, 1H), 3.50-3.46 (m,1H),2.12-2.04(m,1H),1.94-1.86(m,3H)。

Embodiment 21:(S, E)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] Nafoxidine-1-base] synthesis of-4,4,4-trifluoro but-2-ene-1-ketone (compound 21)

With compound 18 prepare similar method, be raw material with trans-4,4,4-trifluorobutene acid, compound can be prepared 21 (64% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.41 (s, 1H), 7.68 (d, J=8.4Hz, 2H),7.46-7.38(m,2H),7.24-7.09(m,5H),6.88-6.82(m,1H),6.78-6.67(m, 1H),5.54-5.52(br,2H),4.78-4.68(m,2H),4.44-4.42(m,1H),3.76-3.67(m, 1H),3.52-3.45(m,1H),2.16-2.08(m,2H),1.78-1.62(m,2H)。

Embodiment 22:(S, E)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] Nafoxidine-1-base] synthesis of-4-oxo but-2-ene acid methyl ester (compound 22)

With compound 18 prepare similar method, with monomethyl fumarate as raw material, compound 22 (70% can be prepared Productivity).¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.23 (s, 1H), 7.64 (dd, J1=8.4Hz, J2 =22.4Hz, 2H), 7.46-7.42 (m, 2H), 7.21-7.12 (m, 5H), 6.69-6.63 (m, 1H), 6.11 (d, J=15.2Hz, 1H), 4.63-4.61 (m, 1H), 4.47-4.43 (m, 1H), 4.29-4.24 (m, 1H), 3.58(s,3H),3.52-3.48(m,2H),2.06-1.98(m,3H),1.86-1.82(m,1H)。

Embodiment 23:(S, Z)-4-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] Nafoxidine-1-base] synthesis of-4-oxo but-2-ene cyanogen (compound 23)

With compound 18 prepare similar method, with maleamic acid as raw material, (64% produces can to prepare compound 23 Rate).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.32 (s, 1H), 7.61 (d, J=6.4Hz, 2H), 7.39-7.36 (m, 2H), 7.15-7.06 (m, 5H), 6.92 (d, J=11.2Hz, 0.5H), 6.82 (d, J= 11.2Hz, 0.5H), 5.76 (d, J=11.2Hz, 0.5H), 5.59 (d, J=11.2Hz, 0.5H), 4.77-4.72(m,1H),4.66-4.50(m,1H),4.42-4.38(m,1H),3.72-3.64(m,1H), 3.48-3.44(m,1H),2.15-1.94(m,4H)。

Embodiment 24:(S, E)-1-[2-[(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) methyl] Nafoxidine-1-base] synthesis of-4-(dimethylamino) but-2-ene-1-ketone (compound 24)

A.4-the synthesis of dimethylamino cronate hydrochlorate

Compound Methyl crotonate (20g, 200mmol) is added, with carbon tetrachloride (200ml) in the single port bottle of 500ml Dissolve, under stirring, add azodiisobutyronitrile (AIBN) (66mg, 0.4mmol) and N-bromo-succinimide (NBS) (39.1g, 220mmol), reactant liquor reacts 6h in being heated to reflux, and cooling, solids removed by filtration, filtrate water washes (20 Ml × 3), anhydrous Na₂SO₄After drying, filtration is spin-dried for obtaining grease 4-bromocrotonic acid methyl ester (34g) of yellow, this change Compound is directly used in next step reaction without purification.

In the single port bottle of 500ml, add compound 4-bromocrotonic acid methyl ester (34g, 190mmol), dissolve with THF, Lower hydrochlorate (18.5g, 230mmol) and the Et adding dimethylamine of stirring₃N (74.2ml, 530mmol), stirs under room temperature Mixing reaction overnight, after 4-bromocrotonic acid methyl ester reacts completely, solvent is evaporated off, remaining yellow oil adds isopropanol (200 Ml) in, HCl/dioxane solution regulation pH≤2 of dropping 4M, separate out white solid, filter, filter cake THF washes Wash, be dried to obtain white solid 4-dimethylamino Methyl crotonate hydrochlorate (10g, 30% productivity).¹H-NMR (d-DMSO, 400MHz, δ ppm): 6.89-6.92 (m, 1H), 6.30 (d, J=16Hz, 1H), 3.91 (d, J=8Hz, 2H), 3.70 (s, 3H),2.72(s,6H)。

In the single port bottle of 250ml, add compound 4-dimethylamino Methyl crotonate hydrochlorate (10g, 56mmol), add The aqueous hydrochloric acid solution (100ml) of 6N, back flow reaction 6h, reactant liquor is spin-dried for obtaining white solid 4-dimethylamino cronate Hydrochlorate (9.0g, 98% productivity).¹H-NMR (d-DMSO, 400MHz, δ ppm): 6.75-6.81 (m, 1H), 6.18 (d, J=16Hz, 1H), 3.89 (d, J=8Hz, 2H), 2.74 (s, 6H).

B. (S, E)-1-(2-((4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) methyl) tetrahydrochysene pyrrole Cough up-1-base) synthesis of-4-(dimethylamino)-2-butylene-1-ketone

With compound 18 prepare similar method, with 4-dimethylamino cronate hydrochlorate as raw material, chemical combination can be prepared Thing 24 (64% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.32 (s, 1H), 7.78-7.67 (m, 2H), 7.36-7.28(m,2H),7.18-7.05(m,5H),6.83-6.77(m,1H),6.55-6.32(m,1H), 4.64-4.58(m,2H),4.38-4.32(m,1H),3.64-3.52(m,2H),3.32-3.18(m,1H), 3.02-2.98(m,1H),2.48(s,6H),2.02-1.74(m,4H)。

Embodiment 25:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of acrylate-2-alkene-1-ketone (compound 25)

A.2-the synthesis of (4-bromobenzene)-2-benzene-1,3-dithiocyclohexane

By 4-Bromophenacyl benzene (10g, 38mmol), dimercaptopropane (5g, 46mmol), p-methyl benzenesulfonic acid (1.1g, 6.4mmol) Be dissolved in toluene (50ml), back flow reaction 12h, be spin-dried for organic solvent, add petroleum ether making beating, filter product (11g, 82% productivity).ESI-MS(m/z):350.9[M+H]⁺。

The synthesis of the most bromo-4-(difluoro (benzene) methyl) benzene

By 2-(4-bromobenzene)-2-benzene-1,3-dithiocyclohexane (5g, 14mmol) is dissolved in anhydrous methylene chloride (50ml) In, room temperature dropping DAST (4ml, 32mmol), drip the reaction of complete stirred overnight, TLC monitors, after reaction terminates, Adding saturated nacl aqueous solution, anhydrous sodium sulfate is dried organic facies, is spin-dried for, and column chromatography purification obtains product (2.8g, 71%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 7.55 (d, J=8.8Hz, 2H), 7.48-7.46 (m, 2H), 7.44-7.41 (m, 3H), 7.37 (d, J=8.8Hz, 2H).¹⁹F-NMR:(CDCl₃,400MHz, δ ppm) :-88.9.

C.4-the synthesis of benzoyl phenylboric acid

Bromo-for 1-4-(difluoro (benzene) methyl) benzene (2g, 7.1mmol) is dissolved in anhydrous THF (20mL), nitrogen Under gas shielded, be cooled to-78 DEG C, dropping n-BuLi (n-BuLi) (3.1mL, 7.8mmol), temperature keep-65 DEG C with Under, after stirring 1h, continue to keep temperature-60 DEG C, dropping triisopropyl borate ester (1.5g, 7.8mmol), slowly rise To 0 DEG C, stir 3h.Add water (10mL) under ice bath to be stirred overnight.Reactant liquor concentrates removes organic facies, uses 12N Concentrated hydrochloric acid PH is adjusted to 1～2, separates out solid, and filtration drying obtains product (1.5g, productivity 95%).ESI-MS(m/z):227.1 [M+H]⁺。

D. (R)-1-(3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) nafoxidine-1-base)-2- The synthesis of propylene-1-ketone

With compound 1 prepare similar method, with 4-benzoyl phenylboric acid and acrylic acid as raw material, can preparing Compound 25 (62% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.40 (s, 1H), 7.98 (d, J= 8.4Hz,2H),7.84-7.80(m,4H),7.65-7.61(m,1H),7.54-7.51(m,2H), 6.62-6.58(m,1H),6.34-6.28(m,1H),5.68-5.62(m,3H),4.96-4.94(m,1H), 4.86-4.84(m,0.5H),4.68-4.64(m,0.5H),4.24-4.20(m,0.5H),4.12-4.08(m, 0.5H),3.74-3.71(m,0.5H),3.41-3.38(m,0.5H),3.22-3.19(m,0.5H), 2.98-2.96(m,0.5H),2.42-2.38(m,2H),2.18-2.02(m,2H)。ESI-MS(m/z):453.2 [M+H]⁺。

Embodiment 26:(R)-[4-[4-amino-1-(1-(vinyl sulfone(RemzaolHuo Xingranliaohuoxingjituan)) piperidines-3-base)-1H-pyrazolo [3,4-d] pyrimidin-3-yl] phenyl] The synthesis of (phenyl) ketone (compound 26)

With compound 25 prepare similar method, with vinylsulfonyl chloride as raw material, compound 26 (53% can be prepared Productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.40 (s, 1H), 7.98 (d, J=8.4Hz, 2H), 7.91-7.84(m,4H),7.64-7.60(m,1H),7.54-7.50(m,2H),6.51-6.44(m,1H), 6.28-6.24 (d, J=16.8Hz, 1H), 6.08-6.03 (m, 1H), 5.72 (br, 2H), 5.06-5.04 (m, 1H),4.03-4.00(m,1H),3.84-3.81(m,1H),3.34-3.31(m,1H),2.72-2.68(m, 1H),2.28-2.22(m,2H),2.20-2.18(m,2H)。

Embodiment 27:(R, E)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-4-oxo but-2-ene cyanogen (compound 27)

A. the synthesis of (E)-methyl 2-cyanoacrylate

(E)-4-amino-4 oxos-2-butylene acid methyl ester (3.18g, 25mmol) is dissolved in pyridine (27ml), under ice bath Dropping phosphorus oxychloride (3.48ml, 36mmol), after stirring 1h, recovers to 1h is stirred at room temperature, and TLC display has been reacted Bi Hou, adds ice cube, and stirring 0.5h, DCM extract 3 times, and organic facies 2M HCl washs, and finally uses NaHCO₃ Washing, organic facies concentrates, and column chromatography purification obtains product (1g, 36% productivity).¹H-NMR(CDCl₃,400MHz,δ : 6.71 (d, J=16.4Hz, 1H), ppm) 6.48 (d, J=16.4Hz, 1H), 3.83 (s, 3H).

The synthesis of b (E)-3-cyano-propenoic acid

(E)-methyl 2-cyanoacrylate (1.4g, 13mmol) is dissolved in THF (5ml), adds methanol (6ml), LiOH (2.28g, 95mmol) and water (2ml).After 10 degrees Celsius of stirring 3h, react complete, add 2M HCl Regulation pH is equal to 3, and ethyl acetate extracts, and organic facies is dried, and is concentrated to give product (945mg, productivity 75%).¹H-NMR (CDCl₃, 400MHz, δ ppm): 9.29 (s, 1H), 6.72 (d, J=16.4Hz, 1H), 6.56 (d, J= 16.4Hz,1H)。

C. (R, E)-4-(3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) nafoxidine-1-

Base) synthesis of-4-oxo-2-butylene cyanogen

With compound 1 prepare similar method, with (E)-3-cyano-propenoic acid as raw material, compound 27 (56% can be prepared Productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.39 (s, 1H), 7.61 (d, J=8.4Hz, 2H), 7.41-7.37 (m, 2H), 7.31-7.19 (dd, J1=16Hz, J2=42Hz, 1H), 7.17-7.14 (m, 3H), 7.08 (d, J=8.4Hz, 2H), 6.49 (d, J=16Hz, 0.5H), 6.43 (d, J=16Hz, 0.5H),5.79-5.72(br,2H),4.90-4.86(m,1H),4.82-4.78(m,0.5H),4.38-4.35(m, 0.5H),4.12-4.06(m,0.5H),3.93-3.89(m,1H),3.52-3.46(m,0.5H),3.32-3.28 (m,0.5H),3.18-3.12(m,0.5H),2.52-2.38(m,1H),2.32-2.22(m,1H), 2.12-2.01(m,1H),1.82-1.68(m,1H)。ESI-MS(m/z):466.6[M+H]⁺。

Embodiment 28:(R)-1-[3-[4-amino-3-(4-(difluoro (phenyl) methyl) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1- Base] piperidin-1-yl] synthesis of acrylate-2-alkene-1-ketone (compound 28)

A.2-the synthesis of (4-(difluoro (phenyl) methyl) phenyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborinate

Bromo-for 1-4-(difluoro (benzene) methyl) benzene (3.76g, 13mmol) is dissolved in dimethoxy-ethane (DME) (100 Ml) and in dimethyl sulfoxide (DMSO) (5ml), under nitrogen protection, [1,1 '-bis-(diphenylphosphine) ferrocene] two is added Palladous chloride. (II) (Pd (dppf) Cl₂) (960mg, 1.3mmol), reacting under the conditions of 80 DEG C overnight, TLC monitors, instead After should terminating, add dichloromethane and water extraction, organic facies anhydrous sodium sulfate is dried, column chromatography purification obtain product (2.1g, Productivity 47.9%).¹⁹F-NMR(400MHz,CDCl₃, δ ppm) :-89.67.¹H-NMR(CDCl₃,400MHz, δ ppm): 7.85 (d, J=8.0Hz, 2H), 7.52-7.47 (m, 4H), 7.43-7.39 (m, 3H), 1.34 (s, 12H)。

B. (R)-1-(3-(4-amino-3-(4-(difluoro (phenyl) methyl) phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidines -1-base) synthesis of-2-propylene-1-ketone

With compound 1 prepare similar method, with 2-(4-(difluoro (phenyl) methyl) phenyl)-4,4,5,5-tetramethyls Base-1,3,2-dioxaborinate and acrylic acid are raw material, can prepare compound 28 (61% productivity).

¹⁹F-NMR(400MHz,CDCl₃, δ ppm) :-89.3.¹H-NMR(CDCl₃,400MHz,δ Ppm): 8.38 (s, 1H), 7.78-7.68 (m, 4H), 7.58-7.54 (m, 2H), 7.45-7.39 (m, 3H), 6.68-6.50(m,1H),6.36-6.28(m,1H),5.73-5.53(m,3H),4.88-4.85(m,1.5H), 4.61-4.52(m,0.5H),4.28-4.22(m,0.5H),4.08-3.96(m,0.5H),3.76-3.68(m, 0.5H),3.36-3.32(m,0.5H),3.21-3.18(m,0.5H),2.94-2.88(m,0.5H), 2.41-2.28(m,2H),2.04-1.92(m,1H),1.88-1.78(m,1H)。ESI-MS(m/z): 475.5[M+H]⁺。

Embodiment 29:(R)-3-[4-(difluoro (phenyl) methyl) phenyl]-1-(1-(vinyl sulfone(RemzaolHuo Xingranliaohuoxingjituan)) piperidines-3-base)-1H-pyrazolo The synthesis of [3,4-d] pyrimidine-4-ammonia (compound 29)

With compound 28 prepare similar method, with vinylsulfonyl chloride as raw material, compound 29 (53% can be prepared Productivity).¹⁹F-NMR(400MHz,CDCl₃, δ ppm) :-89.41.¹H-NMR(CDCl₃,400MHz,δ Ppm): 8.38 (s, 1H), 7.70 (dd, J₁=8.0Hz, J₂=20.4Hz, 4H), 7.54-7.52 (m, 2H), 7.46-7.44(m,3H),6.45-6.42(dd,J₁=16.8Hz, J₂=10.0Hz, 1H), 6.24 (d, J= 16.8Hz, 1H), 6.03 (d, J=10.0Hz, 1H), 5.52-5.49 (br, 2H), 5.04-5.01 (m, 1H), 4.00-3.96(m,1H),3.82-3.79(m,1H),3.28-3.23(m,1H),2.75-2.69(m,1H), 2.23-2.19(m,2H),1.98-1.88(m,2H)。ESI-MS(m/z):511.5[M+H]⁺。

Embodiment 30:[(R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) piperidines-1- Base] synthesis of ((R)-oxirane-2-base) ketone (compound 30)

A. the synthesis of (R)-oxirane-2-carboxylic acid potassium

(R)-glycidic acid methyl ester (1.0g, 9.8mmol) is dissolved in ethanol (8ml) and H₂O (1ml), adds KOH(0.66g,12mmol).Stirring 2 hours under room temperature, completely, solution is directly spin-dried for faint yellow in some board raw material reaction Solid, is directly used in next step.

B. ((R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidin-1-yl) ((R)-ring Oxidative ethane-2-base) synthesis of ketone (compound 30)

By (R)-3-(4-phenoxy phenyl)-1-(piperidines-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia (307mg, 0.79mmol) (R)-oxirane-2-carboxylic acid potassium (200mg, 1.6mmol), 1-ethyl-(3-dimethylaminopropyl) carbodiimide hydrochloride Salt (EDCI) (167mg, 0.87mmol) and I-hydroxybenzotriazole (HOBT) (118mg, 0.87mmol) are dissolved in In DMF (5ml) solvent, it is subsequently adding DIPEA (DIPEA) (307mg, 2.38mmol).42℃ Lower reaction 2 hours, some board raw material disappears.Washing after cooling, ethyl acetate extraction (5ml × 3), organic facies is dried After be spin-dried for, column chromatography purification obtains compound 30 (190mg, productivity 52%).¹H-NMR(CDCl₃,400MHz,δ Ppm): 8.37 (s, 1H), 7.64 (d, J=8.4Hz, 2H), 7.41-7.37 (m, 2H), 7.17-7.14 (m, 3H), 7.08 (d, J=8.4Hz, 2H), 5.50-5.48 (br, 2H), 4.89-4.71 (m, 1H), 4.53-4.37 (m,0.5H),4.34-4.30(m,0.5H),4.22-4.16(m,0.5H),3.90-3.80(m,0.5H), 3.68-3.64(m,1H),3.47-3.41(m,0.5H),3.31-3.20(m,0.5H),3.02-2.98(m,2H), 2.91-2.86(m,1H),2.47-2.34(m,1H),2.28-2.25(m,1H),2.02-1.98(m,1H), 1.80-1.74(m,1H)。

Embodiment 31:[(R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) piperidines-1- Base] synthesis of ((S)-oxirane-2-base) ketone (compound 31)

A. the synthesis of (S)-2-bromo-3-hydracrylic acid

D-Ser (1.0g, 9.5mmol) and KBr (4.0g, 33mmol) stir to the most molten in aqueous solution (8ml), (4.1g, 21mmol, mass concentration is the aqueous solution of 47%, and described mass concentration refers to hydrogen bromide to add HBr under room temperature Quality account for the percentage ratio of aqueous solution of hydrogen bromide gross mass), solution is cooled to-12 DEG C, then by NaNO₂(0.72g,11 Mmol) being slowly added in batches, add solution every time and become sepia, stirring reaction adds again to fading, and goes through 2 hours Adding, recover to 0 DEG C, stirring reaction is overnight.Reactant liquor extracts (5ml × 3) with ether, and organic facies is spin-dried for after drying Obtain oily compound (S)-2-bromo-3-hydracrylic acid (1.0g, productivity 63%).¹H-NMR(CDCl₃,400MHz,δ Ppm): 4.24 (dd, J₁=5.6Hz, J₂=8.0Hz, 1H), 3.80 (dd, J₁=8.0Hz, J₂=11.2Hz, 1H),3.67(dd,J₁=5.6Hz, J₂=11.2Hz, 1H).

B. the synthesis of (S)-oxirane-2-carboxylic acid potassium

(S)-2-bromo-3-hydracrylic acid (1.0g, 5.9mmol) is dissolved in dehydrated alcohol (8ml), and reactant liquor drops to-20 DEG C, Being slowly added dropwise the ethanol solution of KOH (0.63g, 11.3mmol) under nitrogen protection, reactant liquor is at-20 DEG C-0 DEG C Lower reaction is overnight.Rotation is evaporated off most of ethanol, and adding absolute ether (8ml) has solid to separate out, and filters, organic facies Standing has solid to separate out, and again filters, and collects white solid and obtains product (0.3g, productivity 40%).

C. ((R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidin-1-yl) ((S)-epoxy Ethane-2-base) synthesis of ketone

With compound 30 prepare similar method, with (S)-oxirane-2-carboxylic acid potassium as raw material, compound can be prepared 31 (36% productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.37 (s, 1H), 7.66-7.63 (m, 2H), 7.41-7.37(m,2H),7.18-7.14(m,3H),7.09-7.07(m,2H),5.57-5.54(br,2H), 4.89-4.78(m,1.5H),4.58-4.55(m,0.5H),4.05-4.00(m,0.5H),3.88-3.84(m, 0.5H),3.74-3.68(m,1H),3.32-3.26(m,0.5H),3.22-3.15(m,0.5H),2.81-2.72 (m,0.5H),2.44-2.40(m,0.5H),2.38-2.22(m,2H),2.00-1.92(m,1H), 1.78-1.72(m,3H)。

Embodiment 32:(R)-1-[3-[4-amino-3-(4-Phenylsulfanylphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-2-bromine acrylate-2-alkene-1-ketone (compound 32)

A. the synthesis of (4-bromophenyl) (phenyl) sulfane

To bromo thiophenol (5.0g, 26mmol) and iodobenzene (2.7g, 13mmol), tetrabutyl ammonium bromide (TBAB) (4.3g, 13mmol), adds CuI (0.05g, 0.26mmol) in water (50ml) solution of KOH (1.1g, 20mmol), React overnight at 85 DEG C.Iodobenzene fundamental reaction is complete, washing, and ethyl acetate extraction (30ml × 3), organic facies is dried After be spin-dried for, column chromatography purification obtains product (2.8g, productivity 80%).ESI-MS(m/z):264.9[M+H]⁺。

B.4-the synthesis of (thiophenyl) phenylboric acid

Under nitrogen protection, by anhydrous THF (10ml) solution of (4-bromophenyl) (phenyl) sulfane (1.0g, 3.8mmol) Dropping to-70 DEG C, be slowly added dropwise n-BuLi (2.1ml, 4.2mmol), after dripping, stirring reaction 15min, adds boric acid different Propyl ester (0.86g, 4.6mmol), removes the dry ice bath, reacts 30min, solution turned cloudy, have white solid to analyse at 0 DEG C Going out, add water (6ml) after reaction 3h, reaction is overnight.Again add water (8ml), adjust pH=1-2 with dense HCl, EA (8ml × 3) extract, organic facies is spin-dried for after drying, add petroleum ether, be collected by filtration product (0.75g, Productivity 86%).

¹H-NMR (d-DMSO, 400MHz, δ ppm): 8.11 (s, 2H), 7.74 (d, J=8.0Hz, 2H), 7.39-7.35 (m, 4H), 7.23 (d, J=8.0Hz, 2H), 6.56 (s, 1H).

C.1-[(3R)-3-[4-amino-3-(4-Phenylsulfanylphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base]-piperidino]-2-bromine third The synthesis of alkene-1-ketone

With compound 1 prepare similar method, with 4-(thiophenyl) phenylboric acid as raw material, compound 32 (45% can be prepared Productivity).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.37 (s, 1H), 7.60 (d, J=8.4Hz, 2H), 7.48-7.46 (m,2H),7.42-7.34(m,5H),6.04-6.01(s,1H),5.85-5.78(m,1H),5.46-5.38(br,2H), 4.96-4.91(m,1H),4.68-4.62(m,0.5H),4.53-4.47(m,0.5H),4.17-4.11(m,0.5H),3.99-3.94 (m,0.5H),3.79-3.71(m,0.5H),3.52-3.45(m,0.5H),3.22-3.17(m,0.5H),2.93-2.87(m,0.5 H),2.35-2.22(m,2H),2.02-1.98(m,1H),1.80-1.75(m,1H)。

Embodiment 33:(R)-1-[3-[4-amino-3-(4-benzyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2- The synthesis of bromine acrylate-2-alkene-1-ketone (compound 33)

A.4-the synthesis of benzyl phenylboric acid frequency alcohol ester

In 50ml single port flask, addition 4-bromine diphenyl methane (2.5g, 10mmol), connection boric acid pinacol ester (3.885g, 15mmol), KOAc (3.1g, 31mmol), DME (65ml), DMSO (3.3ml).N₂Replace 3 times, add Catalyst Pd (dppf) Cl₂(740mg, 0.5mmol).N₂Replace 3 times, be heated to 80 DEG C, reflux overnight, raw material React.Reactant liquor crosses kieselguhr, adds water, and EA extracts, and organic facies is dried, and concentrates, column chromatography purified product (2.87g, Productivity 97%).ESI-MS(m/z):295.2.

B. the synthesis of (R)-1-Boc-3-(4-amino-3-(4-benzyl phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidines

In 500ml single port bottle, add (R)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidines (2.6g, 5.9mmol), 4-benzyl phenylboric acid frequency alcohol ester (1.2g, 5.9mmol), Na₂CO₃(2.31g, 31mmol), Dimethoxy-ethane (DME) (150ml), H₂O(30ml)。N₂Replace 3 times, add catalyst four triphenylphosphine Palladium (Pd (PPh₃)₄) (0.23g, 0.3mmol), N₂Replace 3 times.Being heated to 80 DEG C, overnight, raw material is anti-in backflow Should be complete.Reaction system is orange settled solution.Rotation, except major part solvent, adds water, and DCM extracts, and organic facies is dried, dense Contracting, column chromatography purification obtains product (2.1g, productivity 75%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.28 (s,1H),7.62-7.57(m,2H),7.54-7.52(m,2H),7.30-7.22(m,2H),7.19-7.14(m,3H), 5.66-5.55(br,2H),4.78-4.73(m,1H),4.32-4.05(m,2H),3.98-4.01(m,2H),3.40-3.26 (m,1H),2.79-2.71(m,1H),2.18-2.11(m,2H),1.82-1.81(m,1H),1.63-1.60(m,1H),1.36 (s,9H)。

C. (R)-1-[3-[4-amino-3-(4-benzyl phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidin-1-yl]-2-bromine acrylate-2- The synthesis of alkene-1-ketone (compound 33)

With compound 1 prepare similar method, with (R)-1-Boc-3-(4-amino-3-(4-benzyl phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) piperidines is raw material, can prepare compound 33 (40% productivity).

¹H-NMR(CDCl₃, 400MHz, δ ppm): 8.36 (s, 1H), 7.60 (d, J=7.6Hz 2H), 7.37-7.31 (m, 4H),7.23-7.22(m,3H),6.03-6.03(m,1H),5.84-5.78(m,1H),5.56-5.49(br,2H),4.93-4.91 (m,1H),4.69-4.65(m,0.5H),4.52-4.49(m,0.5H),4.17-4.11(m,0.5H),4.05(s,2H), 4.01-3.94(m,0.5H),3.79-3.70(m,0.5H),3.48-3.45(m,0.5H),3.18-3.18(m,0.5H), 2.89-2.85(m,0.5H),2.35-2.23(m,2H),1.80-1.59(m,2H)。

Embodiment 34:(R)-1-[3-[4-amino-3-(4-(4-fluorophenoxy) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines -1-base] synthesis of-2-bromine acrylate-2-alkene-1-ketone (compound 34)

A.1-the synthesis of (4-bromobenzene epoxide)-4-fluorobenzene

4-fluorine diphenyl ether (30g, 0.16mol) and several iodine are dissolved in CS₂(300ml), in, it is slowly added dropwise under ice bath Bromine (10ml, 0.19mol), is stirred at room temperature reaction overnight, concentrates, and column chromatography purification obtains product (28g, 64%).¹H-NMR(CDCl₃, 400MHz, δ ppm): 7.46-7.42 (m, 2H), 7.10-6.97 (m, 4H), 6.88-6.84 (m, 2H)。

B.4-the synthesis of (4-fluorophenoxy) phenylboric acid

Being dissolved in anhydrous THF (80mL) by 1-(4-bromobenzene epoxide)-4-fluorobenzene (10g, 37mmol), nitrogen is protected Under, it is cooled to-78 DEG C, dropping n-BuLi (16.4mL, 41mmol), temperature keeps less than-65 DEG C, after stirring 1h, Continue to keep temperature-60 DEG C, dropping triisopropyl borate ester (8.3g, 44.4mmol), be slowly increased to 0 DEG C, stir 3h. Add water (30mL) under ice bath to be stirred overnight.Reactant liquor concentrates removes organic facies, is adjusted to 1～2 with 12N concentrated hydrochloric acid PH, Separating out solid, filtration drying obtains product (6.4g, productivity 75%).ESI-MS(m/z):233.1[M+H]⁺。

C. (R)-1-(3-(4-amino-3-(4-(4-fluorophenoxy) phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base)-piperidino)-2- The synthesis of bromopropene-1-ketone

With compound 1 prepare similar method, with 4-phenoxyphenyl boronic acid as raw material, compound 34 (41% can be prepared Productivity).ESI-MS(m/z):537.1[M+H]⁺。

Embodiment 35:[(R)-3-(4-amino-3-(3-fluoro-4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) tetrahydrochysene pyrrole Cough up-1-base] synthesis of ((S)-oxirane-2-base) ketone (compound 35)

A. the synthesis of (R)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) nafoxidine

By 4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine (10g, 38mmol), (R)-1-Boc-3-hydroxypyrrole (16g, 85mmol), triphenylphosphine (20g, 76mmol) adds in there-necked flask, adds THF (120ml), is cooled to 0 DEG C, Dropping diisopropyl azodiformate (DIAD) (15.2g, 76mmol) and THF (30ml) mixed liquor, about 1h Drip complete, be slowly increased to room temperature reaction overnight.Being spin-dried for by reactant liquor, add water, ethyl acetate extracts, and is dried, and concentrates Rear pillar chromatography purification obtains product (13.8g, yield 84%).ESI-MS(m/z):431.1[M+H]⁺。

The synthesis of the most fluoro-4-phenoxyphenyl boronic acid

With compound 34 prepare similar method, with the bromo-2-of 4-fluoro-1-phenoxy group benzene as raw material, the fluoro-4-of 3-can be prepared Phenoxyphenyl boronic acid.ESI-MS(m/z):233.1[M+H]⁺。

B. (3R)-1-Boc-3-(4-amino-3-(3-fluoro-4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) tetrahydrochysene pyrrole The synthesis coughed up

(R)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) nafoxidine is added in there-necked flask (12.5g, 29mmol), 3-fluoro-4-phenoxyphenyl boronic acid (7.4g, 32mmol), Pd (dppf) Cl₂(0.5g, 0.69 Mmol), sodium carbonate (6.1g, 58mmol), Isosorbide-5-Nitrae-dioxane (160ml) and water (40ml), after nitrogen displacement, It is warming up to 80 DEG C of reactions overnight.After some plate confirms that reaction completes, filtering, be spin-dried for, add water, ethyl acetate extracts, and is dried, Concentrate rear pillar chromatography purification and obtain product (8.8g, productivity 62%).ESI-MS(m/z):491.2[M+H]⁺。

C. the synthesis of (R)-3-(3-fluoro-4-phenoxy phenyl)-1-(pyrroles's-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia

In reaction bulb, (R)-1-Boc-3-(4-amino-3-(3-fluoro-4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1- Base) nafoxidine (7.3g, 14.8mmol) joins trifluoroacetic acid (40ml) and dichloromethane (40ml) mixed solution In, room temperature reaction is overnight.After some plate confirms that reaction completes, use saturated Na₂CO₃Solution is neutralized to after pH is 7-8, use DCM extracts, and organic facies merges, and anhydrous sodium sulfate is dried, and concentrates rear pillar chromatography purification and obtains product (4.96g, yield 86%). ESI-MS(m/z):391.2[M+H]⁺。

D. ((R)-3-(4-amino-3-(3-fluoro-4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base)-1-nafoxidine Base) synthesis of ((S)-oxirane-2-base) ketone (compound 35)

With compound 31 prepare similar method, with (R)-3-(3-fluoro-4-phenoxy phenyl)-1-(pyrroles's-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia is raw material, can prepare compound 35 (43% productivity).ESI-MS(m/z):461.2[M+H]⁺。

Embodiment 36:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine -1-base] synthesis of-2-bromine acrylate-2-alkene-1-ketone (compound 36)

A. the conjunction of (R)-1-Boc-3-(4-amino-3-(4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) nafoxidine Become

(R)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) nafoxidine is added in there-necked flask (12.5g, 29mmol), 4-phenoxyphenyl boronic acid (6.8g, 32mmol), Pd (dppf) Cl₂(0.5g, 0.69mmol), Sodium carbonate (6.1g, 58mmol), Isosorbide-5-Nitrae-dioxane (160ml) and water (40ml), after nitrogen displacement, be warming up to 80 DEG C of reactions are overnight.After some plate confirms that reaction completes, filtering, be spin-dried for, add water, ethyl acetate extracts, and is dried, and concentrates Rear pillar chromatography purification obtains product (8.6g, productivity 63%).ESI-MS(m/z):473.2[M+H]⁺。

B. the synthesis of (R)-3-(4-phenoxy phenyl)-1-(pyrroles's-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia

In reaction bulb, (R)-1-Boc-3-(4-amino-3-(4-phenoxy phenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base) four Hydrogen pyrroles (7g, 14.8mmol) joins in trifluoroacetic acid (40ml) and dichloromethane (40ml) mixed solution, room Temperature reaction is overnight.After some plate confirms that reaction completes, use saturated Na₂CO₃Solution is neutralized to after pH is 7-8, use DCM Extraction, organic facies merges, and anhydrous sodium sulfate is dried, and concentrates rear pillar chromatography purification and obtains product (4.7g, yield 86%).ESI-MS (m/z):373.2[M+H]⁺。

C. (R)-1-(3-(and 4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base]-1-nafoxidine base]-2- The synthesis of bromopropene-1-ketone (compound 36)

By (R)-3-(4-phenoxy phenyl)-1-(pyrroles's-3-base)-1H-pyrazolo [3,4-D] pyrimidine-4-ammonia (184mg, 0.5mmol), 2-bromopropene acid (88mg, 0.58mmol), HOBT (95mg, 0.7mmol) and EDCI (134mg, 0.7mmol), It is dissolved in anhydrous DCM (10ml), adds DIEA (271mg, 2.1mmol), under room temperature, stir 8h.Add second Washing after acetoacetic ester dilution, aqueous phase is extracted with ethyl acetate 2 times again, merges organic facies, and anhydrous sodium sulfate is dried, dense Contracting column chromatography purification obtains product (177mg, productivity 71%).ESI-MS(m/z):505.1[M+H]⁺。

Embodiment 37:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine -1-base] synthesis of-2-chlorine acrylate-2-alkene-1-ketone (compound 37)

With compound 36 prepare similar method, with 2-chloracrylic acid as raw material, (73% produces can to prepare compound 37 Rate).ESI-MS(m/z):461.1[M+H]⁺。

Embodiment 38:(R)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine -1-base] synthesis of-2-fluorine acrylate-2-alkene-1-ketone (compound 38)

With compound 36 prepare similar method, with 2-perfluoroalkyl acrylate as raw material, (73% produces can to prepare compound 38 Rate).ESI-MS(m/z):445.2[M+H]⁺。

Embodiment 39:(R, Z)-4-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] nafoxidine -1-base] synthesis of-4-oxo but-2-ene cyanogen (compound 39)

With compound 36 prepare similar method, with maleamic acid as raw material, (73% produces can to prepare compound 39 Rate).ESI-MS(m/z):452.2[M+H]⁺。

Embodiment 40:[(R)-3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) nafoxidine-1- Base] synthesis of ((R)-oxirane-2-base) ketone (compound 40)

With compound 36 prepare similar method, with (R)-oxirane-2-carboxylic acid potassium as raw material, compound can be prepared 40 (72% productivity).ESI-MS(m/z):443.2[M+H]⁺。

Embodiment 41:(S, E)-1-[3-(4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base) nafoxidine -1-base] synthesis of-3-chlorine acrylate-2-alkene-1-ketone (compound 41)

A. the synthesis of (S)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine-1-base) pyrroles

By 4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine (12g, 46mmol), (S)-1-Boc-3-hydroxypyrrole (19g, 101mmol), triphenylphosphine (24g, 92mmol) adds in there-necked flask, adds THF (120ml), is cooled to 0 DEG C, Dropping DIAD (18.6g, 76mmol) and THF (30ml) mixed liquor, about 1h dropping is complete, is slowly increased to room Temperature reaction is overnight.Being spin-dried for by reactant liquor, add water, ethyl acetate extracts, and is dried, and concentrates rear pillar chromatography purification and obtains product (16 G, yield 81%).ESI-MS(m/z):431.1[M+H]⁺。

B. (S, E)-1-(3-(and 4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-D] pyrimidine-1-base]-1-nafoxidine base]-3- The synthesis of chloropropene-1-ketone

With compound 36 prepare similar method, with (S)-1-Boc-3-(4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine -1-base) pyrroles and the acid of (E)-chlorallylene is raw material, can prepare compound 41 (75% productivity).ESI-MS(m/z): 461.2[M+H]⁺。

Embodiment 42:(S, Z)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] tetrahydrochysene pyrrole Cough up-1-base] synthesis of-3-bromine acrylate-2-alkene-1-ketone (compound 42)

With compound 41 prepare similar method, with (Z)-3-bromopropene acid as raw material, compound 42 can be prepared (71% productivity).ESI-MS(m/z):505.2[M+H]⁺。

Embodiment 43:(R)-1-[3-[4-amino-3-(4-(pyridine-4-epoxide) phenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines -1-base] synthesis of-2-bromine acrylate-2-alkene-1-ketone (compound 43)

A.4-the synthesis of (4-(4,4,5,5-tetramethyl-1,3,2-dioxy boron penta ring-2-base) phenoxy group) pyridine

In 50ml single port flask, add 4-(4-bromobenzene epoxide) pyridine (2.5g, 10mmol), join boric acid pinacol ester (3.9 G, 15mmol), KOAc (3.1g, 31mmol), DME (65ml), DMSO (3.3ml).N₂Replace 3 times, Add catalyst Pd (dppf) Cl₂(740mg, 0.5mmol).N₂Replace 3 times, be heated to 80 DEG C, reflux overnight, Raw material reaction is complete.Reactant liquor crosses kieselguhr, adds water, and EA extracts, and organic facies is dried, and concentrates, column chromatography purified product (2.4 G, productivity 80%).ESI-MS(m/z):298.2.

With compound 1 prepare similar method, with 4-amino-3-iodo-1H-pyrazolo [3,4-D] pyrimidine, 4-(4-(4,4,5,5- Tetramethyl-1,3,2-dioxy boron penta ring-2-bases) phenoxy group) pyridine and 2-bromopropene acid is raw material, can prepare compound 43 (66% productivity).ESI-MS(m/z):520.1[M+H]⁺。

Embodiment 44:(S)-1-[3-[4-amino-3-(4-Phenoxyphenyl)-1H-pyrazolo [3,4-d] pyrimidine-1-base] piperidines-1- Base] synthesis of-2-bromine acrylate-2-alkene-1-ketone (compound 44)

With compound 1 prepare similar method, with 4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine, (R)-1-Boc-3- Hydroxy piperidine and 2-bromopropene acid are raw material, can prepare compound 43 (70% productivity).ESI-MS(m/z): 519.1[M+H]⁺。

Embodiment 45-54: the synthesis of compound 45-54

With compound 1 prepare similar method, with 4-amino-3-iodo-1H-pyrazolo [3,4-d] pyrimidine, (S)-1-Boc-3- Hydroxy piperidine and 2-bromopropene acid are raw material, can prepare compound 45-54.

2. biological activity test part:

Biological Examples 1

Series compound is different growth of tumour cell inhibition tests (CCK8 detection)

One, cell strain:

L1210: mouse lymphotactin leukemia cell line, DMEM+10%FBS；

WSU-DLCL2: human B lymphocyte tumor cell strain, RPMI1640+10%FBS；

The former leukemia cell line of the chronic marrow of K562: people, IMDM+10%FBS；

HL-60: people in loop strain, IMDM+20%FBS；

Two, reagent and consumptive material:

CCK8 test kit；Antitumoral compounds；DMSO.

Three, test method:

1, cell is cultivated

Collect exponential phase cell, counting, with complete medium Eddy diffusion cell,

Adjustment cell concentration, to suitable concn, inoculates 96 orifice plates, and 100 μ l cell suspension are inoculated in every hole.

Cell at 37 DEG C, 100% relative humidity, 5%CO₂Incubator is hatched 24 hours.

2, relative inhibition experiment

Collect exponential phase cell, counting, with complete medium Eddy diffusion cell, adjust cell concentration to the denseest Degree (determining according to cell density optimization Test result), inoculates 96 orifice plates, and every hole adds 100 μ l cell suspension.Cell exists 37 DEG C, 100% relative humidity, 5%CO₂Incubator is hatched 24 hours.

By culture medium, testing compound is diluted to set respective action concentration, adds cell by 25 μ l/ holes.Compound Effect final concentration from 10 μMs to 0 μMs, 4 times of gradient dilutions, totally 10 concentration point.

Cell is placed in 37 DEG C, 100% relative humidity, 5%CO₂Incubator is hatched 72 hours.

Culture medium is abandoned in suction, and the addition complete medium containing 10%CCK-8 is placed in 37 DEG C of incubators hatches 2-4 hour.

On SpectraMax M5Microplate Reader, the absorbance at 450nm wavelength is measured gently after concussion, with At 650nm, absorbance is as reference, calculates suppression ratio.

Data process and result

Be calculated as follows drug on tumor cell growth suppression ratio: growth of tumour cell suppression ratio %= [(Ac-As)/(Ac-Ab)] × 100%

The OA (cell+CCK-8+ testing compound) of As: sample

The OA (cell+CCK-8+DMSO) of Ac: negative control

The OA (culture medium+CCK-8+DMSO) of Ab: positive control

Use software Graphpad Prism 5 and use computing formula log (inhibitor) vs.response to carry out IC₅₀Curve Matching also calculates IC₅₀Value.

In table 1 embodiment, part of compounds is to different growth of tumour cell inhibitory activity (IC₅₀)

Biological Examples 2

Following experiment can be used to measure compound of the present invention, Bruton ' s tyrosine kinase (BTK) enzyme to be lived The inhibitory action of property.

The vitro kinase assay HTRF kinEASE TK kit of Cisbio company, operating procedure reference reagent box description, The method detects the testing compound inhibitory action to BTK enzymatic activity in vitro.Concrete operation step is as follows:

DMSO solution (the DMSO excessive concentration of 2.5% is prepared respectively first by the 1X kinase buffer configured Can on reaction produce impact, control DMSO final concentration of 1%), then with enzyme corresponding 2.5% DMSO solution Dilution testing compound is to corresponding test concentrations (4X).In addition to control wells, in reacting hole used, add the dilute of 4 μ L The testing compound solution released, add in control wells BTK enzyme that 4 μ L had previously prepared corresponding 2.5% DMSO Solution.

The TK-biotin substrate of the BTK enzyme correspondence concentration of substrate that 2 μ L had previously prepared is added in all reacting holes Solution (during enzyme screening, the consumption of substrate is shown in Table 1).

Enzymatic solution (the consumption of enzyme of the corresponding concentration that 2 μ L had previously prepared is added in all reacting holes in addition to negative hole It is shown in Table 1), negative hole supplies volume with 2 μ L enzyme correspondence 1Xkinase buffer.With shrouding film shrouding, after mixing, room temperature is incubated Educate 10 minutes, allow compound and enzyme fully act on combination.

The ATP solution of 2 μ LBTK enzyme corresponding concentration is added to start kinase reaction, the enzyme of BTK in all reacting holes Response time is 25 minutes (ATP concentration corresponding during enzyme screening and response time are shown in Table 1).

Within 5 minutes before kinase reaction terminates, start to prepare BTK and detect liquid.The detection buffer in test kit is used to divide Pei Zhi Streptavidin-XL665 and TK antibody europium cryptate (1:100) of two kinds of enzyme corresponding concentration Detection liquid (detectable concentration corresponding during enzyme screening is shown in Table 1).

After kinase reaction terminates, in all reacting holes, add what 5 μ L had diluted Streptavidin-XL665, mixed Add the TK antibody europium cryptate diluted after even immediately and detect liquid.

Shrouding mixes, and after room temperature reaction 1h, detects fluorescence signal (320nm with ENVISION (Perkinelmer) instrument Stimulating, 665nm, 615nm launch).The suppression ratio in each hole, multiple hole is calculated by full activity hole and background signal hole Average, analyze software PRISM 5.0 with the picture of specialty simultaneously and each testing compound is carried out half inhibitory activity (IC₅₀) matching.

Table 2, the series compound IC to BTK enzyme₅₀Inhibitory activity is tested

Biological Examples 3

Internal pharmacodynamic evaluation

SCID Beige mice, female, 5～6 week old, body weight 18g ± 2g, have purchased from Beijing dimension tonneau China laboratory animal Limit company, SPF level environment is raised.

Taking the logarithm the WSU-DLCL2 cell of trophophase, 1000rpm is centrifuged 5min, and serum-free medium is resuspended, takes 10 μ L Carrying out Trypan Blue living cell counting number (cell survival rate > 90%), adjusting cell density is 1x107/0.2mL. Take 4 5-6 week old SCID mice, in the subcutaneous each injection WSU-DLCL2 cell 1x107 of left and right axil under aseptic condition Individual/side/0.2mL, about two weeks i.e. it can be seen that significantly subcutaneous tumors is formed.When tumor size grows into 300-500mm3, Mice euthanasia, aseptically peels off tumor, is placed in 1640 culture medium, and the tumor mass being cut into 1-2mm3 is (every Individual subcutaneous tumors is about divided into 20-30 tumor mass).Use No. 12 trocars that tumor mass is inoculated into 80 5-6 week old SCID The right axil of mice is subcutaneous.

When tumor-bearing mice tumor grows to measurable size, use SPSS 17.0 software according to mean tumor volume homeostatic principle Mice is randomly divided into 8 groups.COMPOUNDS EXAMPLE 1 and COMPOUNDS EXAMPLE 2 are with 90,30,10mg/kg, and every day fills Stomach is administered；Positive control drug according to Shandong for Buddhist nun with 30mg/kg, every day gastric infusion, administration volume is 0.1ml/10g.Every day Being administered once, successive administration 23 days, negative control group, to equivalent solvent (normal saline solution of 1%DMSO), is administered the phase During recovering, measure weekly Mouse Weight and tumor footpath 2～3 times.Gross tumor volume and Relative tumor is calculated according to measurement data Volume, the computing formula of gross tumor volume (Tumor Volume, TV) is: TV=1/2 × a × b2, wherein a, b represent respectively Tumor major diameter and minor axis.Calculate relative tumour volume (Relative tumor volume, RTV) according to measurement result, calculate Formula is: RTV=Vt/V0, and wherein V0 is gross tumor volume during on-test, and Vt is the gross tumor volume every time measured. The evaluation index of anti-tumor activity is Relative tumor appreciation rate T/C (%), and computing formula is as follows: T/C (%)=TRTV/CRTV × 100%, TRTV is treatment group RTV；CRTV is negative control group RTV, the most swollen Tumor growth inhibition ratio (tumour inhibiting rate) %=(1-T/C) × 100%, result is as shown in table 3.

Table 3: the compounds of this invention therapeutical effect to WSU-DLCL2 Nude Mice

Claims

1. a Pyrazolopyrimidine derivative shown in formula I, its stereoisomer, solvate, pharmaceutically acceptable Salt, active metabolite or prodrug,

Wherein: L be O, S,OrWhen L be O, S,OrTime, Z isOrWhen L isOrTime, Z isOrWherein, X is halogen atom, cyano group,OrR is C₁~ C₄Alkyl；

Ar is substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl；Described " substituted or unsubstituted virtue Base or substituted or unsubstituted heteroaryl " described in " replacement " refer to by one or more selected from halogen atom, cyanogen The substituent group of base, substituted or unsubstituted alkyl, alkoxyl, thiazolinyl, alkynyl and aryl is replaced, and described " replaces Or unsubstituted alkyl " described in " replacement " refer to be replaced by one or more halogen atoms, when there is multiple taking During for base, described substituent group is identical or different；

V is hydrogen atom or fluorine atom；

Y is substituted or unsubstituted alkylidene, cycloalkyl or Heterocyclylalkyl；Described " substituted or unsubstituted alkylidene " Described in replacement refer to by hetero atom be oxygen, sulfur or nitrogen-atoms, hetero atom number be the C of 1-3₄～C₇Heterocyclylalkyl, C₁～C₆Alkoxyl, C₁～C₆Alkylthio group and C₁～C₆Alkylamino in one or more replaced.

2. Pyrazolopyrimidine derivative the most shown in formula I as claimed in claim 1, its stereoisomer, solvate, Pharmaceutically acceptable salt, active metabolite or prodrug, it is characterised in that:

V is hydrogen atom；

And/or,

When X is halogen atom, described halogen atom is fluorine atom, chlorine atom or bromine atoms；

And/or,

When R is C₁～C₄Alkyl time, described " C₁～C₄Alkyl " be methyl, ethyl, propyl group, isopropyl, Butyl, isobutyl group or the tert-butyl group；

And/or,

When the replacement described in " the substituted or unsubstituted alkylidene " described in described Y refer to by hetero atom be oxygen, Sulfur or nitrogen-atoms, hetero atom number are the C of 1-3₄～C₇Heterocyclylalkyl replaced time, described " hetero atom be oxygen, Sulfur or nitrogen-atoms, hetero atom number are the C of 1-3₄～C₇Heterocyclylalkyl " be hetero atom be that nitrogen-atoms, hetero atom number are The C of 1-2₄～C₅Heterocyclylalkyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by When one or more halogen atoms are replaced, described " halogen atom " is fluorine atom, chlorine atom or bromine atoms；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by When one or more substituted or unsubstituted alkyl are replaced, described " substituted or unsubstituted alkyl " for replace or not Substituted C₁～C₄Alkyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by One or more substituted or unsubstituted alkyl are replaced, and taking described in described " substituted or unsubstituted alkyl " When generation refers to be replaced by halogen atom, described halogen atom is fluorine atom, chlorine atom or bromine atoms；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by When one or more alkoxyls are replaced, described " alkoxyl " is C₁～C₄Alkoxyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by When one or more thiazolinyls are replaced, described " thiazolinyl " is C₂～C₄Thiazolinyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by When one or more alkynyls are replaced, described " alkynyl " is C₂～C₄Alkynyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by When one or more aryl are replaced, described " aryl " is C₅～C₁₀Aryl；

And/or,

When described Ar is substituted or unsubstituted aryl, described " substituted or unsubstituted aryl " be " replace or Unsubstituted C₅～C₁₀Aryl "；

And/or,

When described Ar is substituted or unsubstituted heteroaryl, described " substituted or unsubstituted heteroaryl " is miscellaneous former The substituted or unsubstituted C that son is nitrogen-atoms, hetero atom number is 1-2₃～C₉Heteroaryl；

When Y is substituted or unsubstituted alkylidene, described " substituted or unsubstituted alkylidene " is for replacing or not taking The C in generation₁～C₆Alkylidene；

And/or,

When Y is cycloalkyl, described cycloalkyl is C₄～C₇Cycloalkyl；

And/or,

When Y is Heterocyclylalkyl, described Heterocyclylalkyl be hetero atom be oxygen, sulfur or nitrogen-atoms, hetero atom number be 1-3 Individual C₄～C₇Heterocyclylalkyl.

3. Pyrazolopyrimidine derivative the most shown in formula I as claimed in claim 2, its stereoisomer, solvate, Pharmaceutically acceptable salt, active metabolite or prodrug, it is characterised in that:

When the replacement described in " the substituted or unsubstituted alkylidene " described in described Y refers to by hetero atom be that nitrogen is former Son, hetero atom number are the C of 1-2₄～C₅Heterocyclylalkyl replaced time, it is described that " hetero atom is nitrogen-atoms, miscellaneous former Subnumber is the C of 1-2₄～C₅Heterocyclylalkyl " be pyrrolidinyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by One or more substituted or unsubstituted C₁～C₄Alkyl replaced time, described " substituted C₁～C₄Alkyl " For trifluoromethyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by One or more substituted or unsubstituted C₁～C₄Alkyl replaced time, described " unsubstituted C₁～C₄Alkyl " For methyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by One or more C₁～C₄Alkoxyl replaced time, described " C₁～C₄Alkoxyl " be methoxyl group；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by One or more C₂～C₄Thiazolinyl replaced time, described " C₂～C₄Thiazolinyl " be vinyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by One or more C₂～C₄Alkynyl replaced time, described " C₂～C₄Alkynyl " be acetenyl；

And/or,

When the replacement described in described " substituted or unsubstituted aryl or substituted or unsubstituted heteroaryl " refer to by One or more C₅～C₁₀Aryl replaced time, described " C₅～C₁₀Aryl " be phenyl；

And/or,

When described Ar is substituted or unsubstituted C₅～C₁₀Aryl time, described " substituted or unsubstituted C₅～C₁₀ Aryl " be substituted or unsubstituted phenyl；

And/or,

When described Ar be hetero atom be nitrogen-atoms, hetero atom number be the substituted or unsubstituted C of 1-2₃～C₉Miscellaneous During aryl, described " hetero atom be nitrogen-atoms, hetero atom number be the substituted or unsubstituted C of 1-2₃～C₉Heteroaryl Base " be hetero atom be nitrogen-atoms, hetero atom number be the substituted or unsubstituted C of 1₃～C₅Heteroaryl；

And/or,

When Y is substituted or unsubstituted C₁～C₆Alkylidene time, described " substituted or unsubstituted C₁～C₆Asia Alkyl " it is substituted or unsubstituted methylene；

When Y is C₄～C₇Cycloalkyl time, described " C₄～C₇Cycloalkyl " be C₅～C₆Cycloalkyl；

And/or,

When Y be hetero atom be oxygen, sulfur or nitrogen-atoms, hetero atom number be the C of 1-3₄～C₇Heterocyclylalkyl time, described " hetero atom be oxygen, sulfur or nitrogen-atoms, hetero atom number be the C of 1-3₄～C₇Heterocyclylalkyl " be hetero atom be nitrogen Atom, hetero atom number are the C of 1-2₄～C₅Heterocyclylalkyl.

4. Pyrazolopyrimidine derivative the most shown in formula I as claimed in claim 3, its stereoisomer, solvate, Pharmaceutically acceptable salt, active metabolite or prodrug, it is characterised in that:

When described Ar is substituted or unsubstituted phenyl, described " substituted phenyl " is former by one or more fluorine Son substituted phenyl, the phenyl by one or more methyl substituted phenyl, replaced by one or more cyano group, by one Or the multiple substituted phenyl of trifluoromethyl, by the phenyl of one or more methoxy substitutions, taken by one or more vinyls The phenyl in generation, the phenyl replaced by one or more acetenyls or the phenyl being optionally substituted with one or more phenyl groups；

And/or,

When described Ar be for hetero atom be nitrogen-atoms, hetero atom number be the substituted or unsubstituted C of 1₃～C₅Miscellaneous During aryl, described " hetero atom be nitrogen-atoms, hetero atom number be the substituted or unsubstituted C of 1₃～C₅Heteroaryl " For substituted or unsubstituted pyridine radicals；

And/or,

When Y is substituted or unsubstituted methylene, described " substituted methylene " is the substituted methylene of pyrrolidinyl Base；

When Y is C₅～C₆Cycloalkyl time, described " C₆Cycloalkyl " be cyclohexyl；

And/or,

When Y be hetero atom be nitrogen-atoms, hetero atom number be the C of 1-2₄～C₅Heterocyclylalkyl time, described is " miscellaneous former The C that son is nitrogen-atoms, hetero atom number is 1-2₄～C₅Heterocyclylalkyl " be pyrrolidinyl or piperidyl.

5. Pyrazolopyrimidine derivative the most shown in formula I as claimed in claim 4, its stereoisomer, solvate, Pharmaceutically acceptable salt, active metabolite or prodrug, it is characterised in that:

When the phenyl that described Ar is replaced by one or more fluorine atoms, described " is taken by one or more fluorine atoms The phenyl in generation " beOr

And/or,

When described Ar is by one or more methyl substituted phenyl, described " by one or more methyl substituted Phenyl " it is 4-aminomethyl phenyl；

And/or,

When the phenyl that described Ar is replaced by one or more cyano group, described " is replaced by one or more cyano group Phenyl " it is 2-cyano-phenyl；

And/or,

When the phenyl that described Ar is replaced by one or more trifluoromethyls, described " by one or more fluoroforms The substituted phenyl of base " it is 4-trifluoromethyl；

And/or,

When described Ar is by the phenyl of one or more methoxy substitutions, described " is taken by one or more methoxyl groups The phenyl in generation " it is 3-methoxyphenyl；

And/or,

When described Ar is by the phenyl of one or more vinyl substituted, described " is taken by one or more vinyls The phenyl in generation " be

And/or,

When the phenyl that described Ar is replaced by one or more acetenyls, described " is taken by one or more acetenyls The phenyl in generation " be

And/or,

When described Ar is the phenyl being optionally substituted with one or more phenyl groups, described " is optionally substituted with one or more phenyl groups Phenyl " be

And/or,

When described Ar is substituted or unsubstituted pyridine radicals, described " unsubstituted pyridine radicals " is

And/or,

When Y is the substituted methylene of pyrrolidinyl, described " the substituted methylene of pyrrolidinyl " isOr

And/or,

When Y is pyrrolidinyl, described " pyrrolidinyl " isOr

And/or,

When Y is piperidyl, described " piperidyl " isOr

6. Pyrazolopyrimidine derivative the most shown in formula I as claimed in claim 1, its stereoisomer, solvate, Pharmaceutically acceptable salt, active metabolite or prodrug, it is characterised in that:

When L be O, S,OrTime, Z isOr Wherein, X is Cl, Br or CN, and Y is pyrrolidinyl or piperidyl；

And/or,

When L isOrTime, Z is OrWherein, X is Cl, Br or CN, and Y is pyrrolidinyl or piperidyl.

7. Pyrazolopyrimidine derivative the most shown in formula I as claimed in claim 1, its stereoisomer, solvate, Pharmaceutically acceptable salt, active metabolite or prodrug, it is characterised in that:

Described Pyrazolopyrimidine derivative shown in formula I, the compound for as shown in Formulas I-A:

Wherein, the definition of Ar as described in any one of Claims 1 to 5, the definition of L as described in claim 1 or 6, Y Definition as described in any one of claim 1～6, the definition of Z as described in any one of claim 1,2 or 6, V's Definition is as claimed in claim 1 or 2.

8. Pyrazolopyrimidine derivative the most shown in formula I as claimed in claim 7, its stereoisomer, solvate, Pharmaceutically acceptable salt, active metabolite or prodrug, it is characterised in that:

In the present invention, described Pyrazolopyrimidine derivative shown in formula I, for following arbitrary compound:

9. the preparation method of the Pyrazolopyrimidine derivative shown in formula I as described in any one of claim 1～8, its It is characterised by that it comprises the following steps: compound II is carried out coupling reaction with acid ZOH, obtains pyrazoles shown in formula I And pyrimidine derivatives；

10. the arbitrary compound shown in formula I as described in any one of claim 1～8 or its pharmaceutically acceptable salt Or the purposes that its stereoisomer, solvate, active metabolite and prodrug are in preparing medicine, it is individually dosed, Or with other therapeutic agent medication.

11. arbitrary compounds shown in formula I as described in any one of claim 1～8 or its pharmaceutically acceptable salt Or the purposes that its stereoisomer, solvate, active metabolite and prodrug are in preparing medicine, described medicine For treat and/or prevent mammal with compound for suppressing bruton's tyrosine kinase activity or for treating Benefit from the purposes of disease, disease or the condition of illness of the suppression of bruton's tyrosine kinase activity.

Purposes described in 12. claim 10～11, its described suppression bruton's tyrosine kinase activity is used for treating cancer Disease, includes but not limited to entity tumor and hematologic cancers.

Purposes described in 13. claim 10～11, its described suppression bruton's tyrosine kinase activity is used for treating inflammation Disease property disease or morbid state, immunity class disease or morbid state, hyperplasia class disease or morbid state and degeneration class Disease or morbid state.

14. 1 kinds of pharmaceutical compositions, it comprises reactive compound and one or more pharmaceutically acceptable carrier or tax Shape agent；Described reactive compound is the Pyrazolopyrimidine derivative shown in formula I described in any one of claim 1～8, One or many in its stereoisomer, solvate, pharmaceutically acceptable salt, active metabolite and prodrug Kind.

15. pharmaceutical compositions as claimed in claim 14, it is characterised in that: the dosage form of described pharmaceutical composition includes But be not limited to aqueous liquid dispersion, self emulsifying dispersion, solid solution, liposomal dispersion agent, aerosol, solid dosage forms, Powder, immediate release formulations, controlled release preparation, dissolution formulation, tablet, capsule, pill, slow releasing preparation, delayed release dosage system, Instant and the controlled release preparation of pulsation delivery formulations, many granular preparations and mixing.