CN105399686B

CN105399686B - Pyrimidine derivatives, its preparation method and its application

Info

Publication number: CN105399686B
Application number: CN201410472716.1A
Authority: CN
Inventors: 鲁先平; 余金迪; 杨千姣; 李志斌; 潘德思; 山松; 朱江飞; 王祥辉; 刘湘衡; 宁志强
Original assignee: SHENZHEN WEIXIN BIOLOGICAL SCIENCE AND TECHNOLOGY Co Ltd
Current assignee: Shenzhen microbiology Polytron Technologies Inc
Priority date: 2014-09-16
Filing date: 2014-09-16
Publication date: 2018-05-22
Anticipated expiration: 2034-09-16
Also published as: CN105399686A

Abstract

The present invention provides shown in formula (I) pyrimidine derivatives or its pharmaceutically acceptable salt, its preparation method and its application.Pyrimidine derivatives or its pharmaceutically acceptable salt shown in formula (I) have jak kinase inhibitory activity, especially inhibitory activity selective to JAK3 kinases, higher, it can be used for preparing JAK3 kinase inhibitors, prevention or treatment and the drug of the relevant disease of JAK3 abnormal kinases are used to prepare, so as to prevent or treat and the relevant disease of JAK3 abnormal kinases.

Description

Pyrimidine derivatives, its preparation method and its application

Technical field

The present invention relates to technical field of pharmaceuticals, and in particular to a kind of pyrimidine derivatives, its preparation method and its application.

Background technology

2002, Manning et al. determined that human kinase group includes 518 kinds of protein kinase genes, wherein 218 kinds of enzymes Occurrence and development closely related (Manning G., the et al.2002, Science, 298 of gene and human diseases:1912- 1934).In the drug having now been found that, the drug using enzyme as action target spot accounts for as many as 20%, is especially targeted protein kinase There is special value in clinical practice.

Protein kinase is that a kind of intracellular messenger relies on, is catalyzed specific protein phosphorylation and completes signal transduction process Enzyme, mainly including tyrosine protein kinase, such as JAKs, Src, Abl, EGFR, FGFR, PDGFR；Serine/threonine protein Kinases, such as PKC, MAPK, Rho kinases；Dual specificity protein kinases, such as MAPKK and phosphatidyl inositol kinase, such as PI3K.Protein kinase phosphorylation/dephosphorylation process can adjust the various biological process of different cells, such as metabolism, cell (the Shchemelinin I., et such as differentiation, cell survival, Apoptosis, orga- nogenesis, angiogenesis, immune response al.2006,Folia Biol.,52:81-100)。

JAKs (Janus kinase) is non-receptor in endochylema including four known member JAK3, JAK1, TYK2, JAK2 A Ge little families in tyrosine protein kinase superfamily.JAK3 is distributed in marrow and lymphatic system, and latter three is then divided extensively It is distributed in Various Tissues cell.After different cytokines combination cell surface receptor, the JAKs of activated receptor coupling, and then make Receptor phosphorylation, this is endochylema signal transduction and activating transcription factor stat protein (Signal Transducers and Activators of Transcription, STAT1~4, STAT5a, STAT5b, STAT6) recruitment site is provided, JAKs phosphorylation stat proteins are transferred to controlling gene in nucleus after the latter's dimerization and express, this approach, that is, JAK/STAT Signal path (O ' Shea J.J., et al.2013, N.Engl.J.Med., 368:161-170).

JAK/STAT signal paths are a signal transduction pathways stimulated by cytokine profiles and growth factor receptors, These factors include interleukin class (IL-2~7, IL-9, IL-10, IL-15, IL-21), interferons (IFN-α, IFN-β, IFN-γ), hematopoietin (EPO), granulocyte and giant cell colony stimulating factor (GM-CSF), somatotropin (GH), Prolactin (PRL), thrombopoietin (TPO) etc. are participating in multiplication, the immunological regulation of immunocyte and candidate stem cell Biological process in play a crucial role (Ghoreschi K., et al.2009, Immunol.Rev., 228:273-287).No Isoacceptor can activate the jak kinase of different subtype, so as to fulfill the biological function of differentiation.

JAK3 with the γ in the cytokine receptor complex such as IL-2, IL-4, IL-7, IL-9, IL-15, IL-21 by being total to Chain (γ c) is combined, and adjusts cellular signal transduction.JAK3 or γ c mutation can all cause severe combined immunodeficiency (SCID) (Villa A.,et al.1996,Blood,88:817-823).JAK3 activity Novel presentation largely subtracts for T cell and NK cells Less, B cell function is lost, and seriously affects the natural biological function of immune system etc..Based on its functional characteristics and special group Distribution is knitted, JAK3 becomes for the very attractive drug target of immune system relevant disease, and inhibitor is closed in rheumatoid Section scorching (RA), Crohn disease, systemic loupus erythematosus, multiple sclerosis, Type I diabetes, psoriasis, anaphylactia, heavy breathing Have in terms of asthma, chronic obstructive pulmonary disease, leukaemia, lymthoma, organ transplant and the treatment of the diseases such as other/prevention important Clinical value (Papageorgiou A.C., et al.2004, Trends Pharm.Sci., 2004,25:558-562).

JAK1 can in IL-10, IL-19, IL-20, IL-22, IL-26, IL-28, IFN-α, IFN-γ, gp130 families IL-6 and other receptors etc. of the c containing γ combine (Scott J.R., et al.1998, Cell, 93:373-383).Mouse mould JAK1 gene knockout experiments in type show that the enzyme plays pass in the biological effect for adjusting above-mentioned cytokine profiles receptor Key acts on (Kisseleva T., et al.2002, Gene, 285:1-24).JAK1 be immunized, the disease areas such as inflammation and cancer New target spot.JAK1 inhibitor can be used for treating/prevention of autoimmune diseases, inflammation and tumour (Hornakova T., et al.2010,Blood,115:3287-3295), as leukaemia, lymthoma, melanoma, arthritis, psoriasis, Crohn disease, Lupus erythematosus, Immune Deficiency Syndrome, Behcet's disease (Hou S., et al.2013, Hum.Genet., 132:1049- 1058) etc., but it is not limited to this.

TYK2 is first member in JAK families, can be by IFNs, IL-10, IL-6, IL-12, IL-23, IL-27 etc. A variety of receptor activations.In mouse, TYK2 afunction can cause the signal path of cytokine profiles receptor that defect occurs, into And cause virus infection, antibacterial immunity function reduction and add (Kisseleva T., the et such as possibility of pulmonary infection al.2002,Gene,285:1-24).In addition, the research of Larner A.C. groups shows that TYK2 can help to inhibit breast cancer Growth and transfer (Zhang Q., et al.2011, J.Interferon Cytokine Res., 31:671-677)；The problem Group is it has recently been reported that TYK2 can assist grinding for fat adjusting in mouse and human body by the differentiation of brown adipose tissue (BAT) Study carefully achievement, body can be protected from obesity or even obesity (Derecka M., et al.2012, Cell can be reversed Metab.,2012,16:814-824).This perhaps provides new chance for fat cancer patient's treatment.

JAK2 plays a significant role during being adjusted including a variety of receptor signals such as EPO, GH, PRL, IL-3, IFN-γ (Kisseleva T.,et al.2002,Gene,285:1-24；Levy D.E.,et al.2002,Nat.Rev.Mol.Cell Biol.,3:651-662；O’Shea J.J.,et al.2002,Cell,109(suppl.):S121-S131).In mouse model Animal dead (Schindler C., et al.2007, J.Biol.Chem., 282 caused by middle knockout JAK2 can cause anaemia: 20059-20063)；A base mutation JAK2V617F on JAK2 genes in human body, and in myeloproliferative disease Polycythemia vera (PV), essential thrombocythemia (ET), idiopathic myelofibrosis (IMF), chronic granulocyte Generation closely related (Ghoreschi K., the et al.2009, Immunol.Rev., 228 of leukaemia (CML) etc.:273- 287).Therefore, JAK2 has become the definite action target spot for the treatment of/prevention of such disease.

In November, 2012, the pan-JAKs inhibitor Xeljanz (Tofacitinib) of Pfizer companies have obtained FDA approvals For treating RA.In October, 2013, the said firm disclose Xeljanz for treating III clinical data of psoriasis, control The double-blind trial of Enbrel (Etanercept), the drug meet Noninferior solution design studies result.However, Xeljanz has Have including causing the side effects such as red blood cell and quantity of leucocyte decline, cholesterol levels rise, this is perhaps with it with high JAK2 Inhibitory activity correlation (Zak M., et al.2012, J.Med.Chem., 55:6176-6193).Therefore, major drugmaker etc. The scientist of mechanism focuses on sight in the research and discovery of selective JAK inhibitor, such as selective JAK 3 restrainer, Selective JAK1 inhibitor etc..

It there is now multiple choices JAK inhibitor and be in different clinical investigation phases, including selective JAK 3 restrainer VX-509, selectivity JAK1 inhibitor GLPG0634 (Feist E., et al.2013, Rheumatology, 52:1352- And INCB39110 (http 1357)://www.incyte.com/research/pipeline) etc., for treating RA, Crow grace The immune systems relevant disease such as disease, psoriasis, myelofibrosis.In addition, the selective depression of a variety of different types of structure The related patents of agent are disclosed：1) selective JAK 3 restrainer, such as pyrrolo- [1,2-b] pyridazine (WO2012125887), pyrazoles And [3,4-d] pyrimidine (WO2011048082, WO2011134861, WO2012022681), diaminopyrimidine (WO2011029807, WO2012015972), pyrrolo- [2,3-b] pyridine (JP2012012332), diamino-pyridine -3- first Amide (WO2010061971, US20120108566), pyrrolo- [2,3-b] pyrazine (WO2011144584, WO2011144585) Deng；2) selectivity JAK1 inhibitor, such as tricyclic compounds (WO2011086053), the pyrazoles of substitution and pyroles (WO2010135650, WO2011112662), anilino- phthalazines class (WO2012037132) etc..In addition, with selectivity JAK2 Inhibitor, selectivity TYK2 inhibitor and simultaneously tool there are two types of hypotype (JAK3/1, JAK1/2 etc.) inhibitor related patents Also it has been reported, details are not described herein.

The content of the invention

One of the objects of the present invention is to provide a kind of pyrimidine derivatives, with selective JAK 3 kinase inhibiting activity；

The second object of the present invention is to provide a kind of preparation method of pyrimidine derivatives；

The third object of the present invention is in the application in offer pyrimidine derivatives in JAK3 kinase inhibitors are prepared.

The fourth object of the present invention is to provide pyrimidine derivatives to prepare treatment relevant with JAK3 abnormal kinases Application in the drug of disease.

The present invention provides the pyrimidine derivatives or its pharmaceutically acceptable salt shown in formula (I)：

Wherein,

A is selected from 6~10 yuan of aromatic rings or 5~10 yuan of hetero-aromatic ring；

X is halogen；

R¹Independently selected from hydrogen, cyano, halogen, amino, hydroxyl, the alkyl of C1~C6 or halogenated alkyl, the alkane of C1~C6 One or more of oxygroup；

R²Independently selected from one or more of hydrogen or halogen；

R³For hydrogen or the alkyl of C1~C6 or substitution alkyl；

N is 1 to 7 integer；

P is 0 to 3 integer.

Present invention additionally comprises the enantiomter of compound, diastereoisomer, hydrate, solvates shown in formula (I) Deng preferably including stereoisomer or hydrate.

In the present invention, A is preferably phenyl ring or pyridine ring selected from 6~10 yuan of aromatic rings or 5~10 yuan of hetero-aromatic ring.

In the present invention, X is halogen, is preferably chlorine.

In the present invention, R¹Independently selected from hydrogen, cyano, halogen, amino, hydroxyl, the alkyl of C1~C6 or halogenated alkyl, One or more of alkoxy of C1~C6 is more preferably selected from the halogenated alkyl of hydrogen, cyano, halogen, amino, C1~C6 One or more, R¹For it is multiple when, may be the same or different, be preferably difference；R¹Number be preferably 1~3, more Preferably 1~2.Wherein, halogenated alkyl is preferably the alkyl of fluorine substitution, and the number of fluoro substituents can be multiple；Alkyl halide The carbon atom of base is preferably 1~4, more preferably 1~3.

In the present invention, R²It is preferably hydrogen or F independently selected from one or more of hydrogen or halogen.

In the present invention, R³It is preferably hydrogen or methyl for hydrogen or the alkyl of C1~C6 or substitution alkyl.

In the present invention, n and p is the number of methylene, and n is 1 to 7 integer, is preferably 2~4, more preferably 2 or 4；p For 0 to 3 integer.

Preferably, in formula (I), A is phenyl ring or pyridine ring；R¹Independently selected from hydrogen, cyano, fluorine, amino, trifluoromethyl One or more of；R²For hydrogen or halogen；R³For hydrogen or methyl；N is 1 to 4 integer；P is 0 to 2 integer.

Preferably, in formula (I), A is phenyl ring or pyridine ring；R¹Independently selected from hydrogen, cyano, fluorine, amino, trifluoromethyl One or more of；R²For hydrogen or fluorine；R³For hydrogen or methyl；N is 1 to 4 integer；P is 0 to 2 integer.

Preferably, in formula (I), A is phenyl ring；R1 is independently selected from one or more of fluorine, amino, trifluoromethyl； R²For fluorine；R³For methyl；N is 4, p 0.

" halogen " of the present invention is fluorine, chlorine, bromine, iodine；

" alkyl " of the present invention, including straight chain, branch or cyclic alkyl, as methyl, ethyl, n-propyl, isopropyl, Normal-butyl, isobutyl group, tertiary butyl, n-pentyl, isopentyl, n-hexyl, isohesyl, cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl Deng.

The present invention also provides the preparation method of above-mentioned pyrimidine derivatives, including：

Logical formula (VI) compound is formed general formula under organic solvent and catalyst action with logical formula (VII) compound is (I) compound；

Wherein,

R²Independently selected from one or more of hydrogen or halogen；

R³For hydrogen or the alkyl of C1~C6 or substitution alkyl；

N is 1 to 7 integer；

P is 0 to 3 integer.

Logical formula (VI) compound obtains the pyrimidine derivatives shown in formula (I), reaction process with logical formula (VII) compound condensation It is as follows：

The condensation reaction is preferably using peptide condensing agent as catalyst, such as 1- ethyls -3- (3- dimethylamine propyls) carbodiimide (EDC), N, N '-dicyclohexylcarbodiimide (DCC), N, N '-phosphinylidyne diimidazole (CDI) etc..Reaction temperature is preferably 0~60 DEG C, when the reaction time is preferably 2~72 small.Reaction solvent for use is common solvent, such as benzene, toluene, tetrahydrofuran, dioxy six Ring, dichloromethane, chloroform, N,N-dimethylformamide etc..If necessary, alkali such as sodium hydroxide, triethylamine or pyrrole can also be added in Pyridine.

In the present invention, leading to formula (VII) compound directly can commercially obtain, and it is preferred to lead to formula (VI) compound It prepares in accordance with the following methods：

Logical formula (IV) compound is formed general formula under the action of organic solvent and catalyst with logical formula (V) compound is (VI) compound；

With logical formula (V) compound substitution reaction occurs for logical formula (IV) compound, obtains logical formula (VI) compound, reaction process It is as follows：

In the substitution reaction, the catalyst is selected from trifluoroacetic acid, hydrochloric acid or methanesulfonic acid, is preferably trifluoroacetic acid；Reaction Temperature is reflux, when the reaction time is 8-16 small.Common solvent used in reaction is isopropanol, n-butanol etc..

Logical formula (IV) compound can be prepared in accordance with the following methods：

With logical formula (III) compound substitution reaction occurs for logical formula (II) compound, obtains logical formula (IV) compound, reacted Journey is as follows：

Wherein, logical formula (II) compound and logical formula (III) compound are the raw material that market can be directly commercially available, Substitution reaction occurs in the presence of base.Reaction temperature is reflux, when the reaction time is 8-16 small.Common solvent is used in reaction Ethyl alcohol, methanol, n-butanol etc..Alkali is triethylamine, diisopropyl ethyl amine etc..

Logical formula (V) compound can be prepared in accordance with the following methods：

Logical formula (VIII) compound is condensed to yield logical formula (IX) chemical combination with acryloyl chloride in the presence of diisopropyl ethyl amine Object obtains logical formula (V) compound after carrying out reduction.

Logical formula (VIII) compound directly can be obtained commercially, with acryloyl chloride in diisopropyl ethyl amine In the presence of be condensed to yield logical formula (IX) compound, logical formula (IX) compound obtains logical formula (V) compound through iron powder reducing nitro, reacts Process is as follows：

After logical formula (VI) compound obtains the pyrimidine derivatives shown in formula (I) with logical formula (VII) compound condensation, it can adopt Purified with common separation method, such as extraction, recrystallization, column chromatography, the present invention have no this specifically limited.

Specifically, the pyrimidine derivatives shown in formula (I) can be following compound：

1 particular compound of the present invention of table

The present invention also provides a kind of pharmaceutical preparation, including the pyrimidine derivatives described in above-mentioned technical proposal or its pharmaceutically may be used The salt of receiving and pharmaceutically acceptable auxiliary material.

The pharmaceutically acceptable auxiliary material includes carrier, diluent, filler, disintegrant etc., can be according to dosage form Corresponding auxiliary material is selected, including《Pharmaceutical excipient handbook》Carrier filler listed by (American Pharmaceutical Association, in October, 1986), but It is not limited to these carrier fillers.

The drug directly can be the abilities such as tablet, capsule, pulvis, syrup, liquor, suspending agent, injection or paste Dosage form known to field technique personnel, the present invention are not particularly limited its processing method.In preparation, the general formula as active component (I) content of compound is preferably 0.5~70wt%, and preferable content is 1~20wt%.The dosage form specification of the preparation is preferred For 0.0001~200mg.

Pyrimidine derivatives or its pharmaceutically acceptable salt shown in formula (I) have jak kinase inhibitory activity, especially right The selective inhibitory activity of JAK3 kinases can be used for preparing selective JAK 3 kinase inhibitor, be used to prepare prevention or Treatment and the drug of the relevant disease of JAK3 abnormal kinases, it is relevant with JAK3 abnormal kinases so as to prevent or treat Disease, such as autoimmune disease, diseases associated with inflammation, cancer, myeloproliferative disease, bone-resorbing disease or organ transplant Repel disease etc., especially rheumatoid arthritis, Crohn disease, systemic loupus erythematosus, multiple sclerosis, I type glycosuria Disease, psoriasis, anaphylactia, asthma, chronic obstructive pulmonary disease, leukaemia or lymthoma.

Pyrimidine derivatives or its pharmaceutically acceptable salt shown in formula (I) of the present invention, can clinically lead to It crosses oral or injection system and medication is carried out to mammal (including people), wherein especially optimal with oral way.Dosage is every Day 0.0001~200mg/kg weight, preferable dosage are daily 0.01~100mg/kg weight, and optimal dosage is every Day 0.1~50mg/kg weight, meanwhile, depending on optimal dose regards individual, usually beginning when dosage it is smaller, be then gradually increased use Amount.

The experimental results showed that pyrimidine derivatives provided by the invention have higher inhibitory activity to JAK3 kinases, and it is right JAK2 kinases and JAK1 kinase inhibitory activities be not notable.

Based on the above, selective JAK 3 restrainer disclosed herein is used to treating/preventing RA, psoriasis, Crow It is grace disease, systemic loupus erythematosus, multiple sclerosis, Type I diabetes, anaphylactia, chronic obstructive pulmonary disease, asthma, white The immune systems relevant diseases such as blood disease, lymthoma, but it is not limited to this.At the same time, it is desirable to these compounds or its active ingredient Pharmaceutical composition etc. the clinical efficacy of these diseases can be reached to maximization and other purposes in safe treatment window.

Description of the drawings

Fig. 1 is kinase activity testing principle schematic diagram of the embodiment of the present invention.

Specific embodiment

With reference to the example content that the present invention is furture elucidated, but protection scope of the present invention is not limited only to this A little examples.Percentage of the present invention is weight percentage in addition to especially indicating.Numerical value model described in this description It encloses, such as measurement unit, reaction condition, compound physical state or percentage, is for providing undoubted desk reference. Those skilled in the art when putting into practice this patent, using outside this scope or be different from the temperature of single number, concentration, Quantity, carbon number etc. still can obtain expected result.

The preparation of the fluoro- N- methyl-5-nitros aniline of 1 2- of embodiment

The fluoro- 5- nitroanilines (20.0g, 128.2mmol) of 2- and paraformaldehyde (16.0g, 533.3mmol) are dissolved in 500mL It is stirred at room temperature in methanol, the methanol solution 100mL of NaOMe (3.4g, 63mmol) is then added dropwise, the reaction solution is at room temperature Stir 16 it is small when after be divided to two deciles add in NaBH₄(9.7g, 255.2mmol) is stirred 15 minutes, LCMS tracking reactions.Reaction knot It is poured into after beam in the aqueous solution of the KOH of 1M, solid is precipitated in stirring, filters to obtain the fluoro- N- methyl-5-nitros aniline of yellow solid 2- (19.0g, 87%yield).LC-MS(m/z)171(M+1).

The preparation of 2 N- of embodiment (the fluoro- 5- nitrobenzophenones of 2-)-N methacrylamide

The fluoro- N- methyl-5-nitros aniline (11.0g, 64.7mmol) of 2- and DIPEA (23mL, 129.4mmol) are dissolved in It is stirred at room temperature in 100mL THF, acryloyl chloride (11mL, 129.4mmol) is then added dropwise, which is stirred at room temperature 1 it is small when after evaporate most of solvent, add in the dilution of 100mL ethyl acetate, saturated common salt water washing is dry, filters, vacuum distillation Obtain yellow oil N- (the fluoro- 5- nitrobenzophenones of 2-)-N methacrylamide (12.0g, 83%yield).LC-MS(m/z)225 (M+1)。

The preparation of 3 N- of embodiment (5- amino -2- fluorophenyls)-N methacrylamide

Iron powder (20.0g, 357mmol) and NH₄Cl (20.0g, 374mmol), which is dissolved in 200mL water, is heated to 80 DEG C of stirrings Then half an hour adds in the ethyl acetate solution of N- (the fluoro- 5- nitrobenzophenones of 2-)-N methacrylamide (12g, 53.6mmol) 20mL.The reaction solution stirred at 80 DEG C 1 it is small when after add in NaHCO₃Aqueous solution alkali tune filters iron cement, filtrate ethyl acetate Extraction, merge it is organic subtract each other pressure distill brown oil N- (5- amino -2- fluorophenyls)-N methacrylamide (5.6g, 54%yield).LC-MS(m/z)195(M+1).

The preparation of 4 3- of embodiment (2,5- dichloro pyrimidine base -4- amino) propionic acid

By 2,4,5- trichloropyrimidines (1.0g, 5.6mmol), alanine (500mg, 5.6mmol) and triethylamine (0.2mL, 1.4mmol) be dissolved in 25mL ethyl alcohol, the reaction solution be heated to 75 DEG C of stirrings 3 it is small when after react and terminate.Ether is used after solvent evaporated Washing, filters to obtain white solid 3- (2,5- dichloro pyrimidine base -4- amino) propionic acid (1.1g, 83%yield).LC-MS(m/z) 236(M+1)。

5 3- of embodiment (the chloro- 2- of 5- (the fluoro- 3- of 4- (N methacrylamide base) phenyl amino) pyrimidine radicals -4- amino) third The preparation of acid

3- (2,5- dichloro pyrimidine base -4- amino) propionic acid (1.1g, 4.7mmol), N- (5- amino -2- fluorophenyls)-N- first Base acrylamide (1.0g, 5.2mmol) and trifluoroacetic acid (0.5mL, 3.5mmol) are dissolved in 16mL water and 4mL isopropanols.This is anti- Answer liquid be heated to 95 DEG C stirring 4 it is small when after add in NaHCO₃Aqueous solution tune PH to 6-7 has solid precipitation.Solid use is obtained by filtration Ethyl alcohol recrystallization obtains gray solid 3- (the chloro- 2- of 5- (the fluoro- 3- of 4- (N methacrylamide base) phenyl amino) pyrimidine radicals -4- ammonia Base) propionic acid (350mg, 18%yield).

1H NMR (DMSO-d6) δ 2.56 (t, J=7.1Hz, 2H, CH2), 3.59-3.62 (m, 2H, CH2), 3.12 (s, 3H, CH3), 5.61 (d, J=10.2Hz, 1H, CH), 6.03-6.09 (m, 1H, CH), 6.16-6.21 (m, 1H, CH), 7.21 (s, 1H, Ar-H), 7.24-7.28 (m, 1H, Ar-H), 7.71 (t, J=3.4Hz, 1H, miazine-NH), 7.79-7.82 (m, 1H, Ar-H),7.99(s,1H,miazine-H),9.47(s,1H,benzene-NH),12.30(s,1H,COOH).LC-MS(m/z) 394(M+1)。

6 N- of embodiment (5- (4- (3- (4- cyanobenzyls amino) -3- oxopropyls amino) -5- chlorine pyrimidine radicals -2- ammonia Base) -2- fluorophenyls)-N methacrylamide preparation

By 3- (the chloro- 2- of 5- (the fluoro- 3- of 4- (N methacrylamide base) phenyl amino) pyrimidine radicals -4- amino) propionic acid (200mg, 0.51mmol), 4- (amine methyl) cyanophenyl hydrochloride (110mg, 0.65mmol) are dissolved in 10mLDMF, are then added in DIPEA (0.3mL, 1.7mmol), I-hydroxybenzotriazole (103mg, 0.76mmol) and 1- ethyls-(3- dimethylaminos third Base) phosphinylidyne diimmonium salt hydrochlorate (200mg, 1.05mmol), mixture be stirred at room temperature 20 it is small when, then use NaHCO₃It is water-soluble Liquid tune pH value is to 8-10.There is solid precipitation, gray solid N- (5- (4- (3- (4- cyanobenzyls amino) -3- oxos are obtained by filtration Propylcarbamic) -5- chlorine pyrimidine radicals -2- amino) -2- fluorophenyls)-N methacrylamide (220mg, 79%yield).1HNMR (DMSO-d6) δ 2.54 (t, J=6.6Hz, 2H, CH2), 3.18 (s, 3H, CH3), 3.64-3.66 (m, 2H, CH2), 4.36 (d, J =5.7Hz, 2H, CH2), 5.60 (d, J=10.1Hz, 1H, CH), 6.04-6.10 (m, 1H, CH), 6.18-6.22 (m, 1H, ), CH 7.18-7.22 (m, 1H, Ar-H), 7.21 (t, J=10.0Hz, 1H, NH), 7.41 (d, J=8.0Hz, 2H, Ar-H), 7.71 (s, 1H, Ar-H), 7.74 (d, J=8.1Hz, 2H, Ar-H), 7.85 (d, J=5.4Hz, 1H, Ar-H), 7.97 (s, 1H, ), miazine-H 8.52 (t, J=5.7Hz, 1H, miazine-NH), 9.44 (s, 1H, benzene-NH) .LC-MS (m/z) 508 (M+1)。

7 3- of embodiment (2- (1H- benzos [d] [1,2,3] triazole -1- oxygroups) -5- chlorine pyrimidine radicals -4- amino)-N- The preparation of (tri- fluorin benzyl amines of 2,3,6-) propionamide

3- (2,5- dichloro pyrimidine -4- amino) propionic acid (0.5g, 2.1mmol) and 2,3,6- tri- fluorin benzyl amines (0.34g, It 2.1mmol) is dissolved in 10mL DMF, EDC (0.8g, 4.2mmol), HOBt (0.35g, 2.5mmol) is sequentially added, in room temperature Under be stirred overnight.It is diluted with water, solid is precipitated, filters to obtain yellow solid 3- (2- (1H- benzos [d] [1,2,3] triazole -1- oxygen Base) -5- chlorine pyrimidine radicals -4- amino)-N- (tri- fluorin benzyl amines of 2,3,6-) propionamide (0.16g, 16%yield).LC-MS(m/z) 478(M+1)。

8 N- of embodiment (5- (4- (3- (2,3,6- trifluoro-benzyls amino) -3- oxopropyls amino) -5- chlorine pyrimidine radicals -2- Amino) -2- fluorophenyls)-N methacrylamide preparation

By 3- (2- (1H- benzos [d] [1,2,3] triazole -1- oxygroups) -5- chlorine pyrimidine radicals -4- amino)-N- (2,3,6- Three fluorin benzyl amines) propionamide (0.16g, 0.34mmol), N- (5- amino -2- fluorophenyls)-N methacrylamide (0.07g, 0.34mmol) it is dissolved in trifluoroacetic acid (40 μ L, 0.5mmol) in 5mL isopropanols.The reaction solution be heated to 95 DEG C stirring 4 it is small when Afterwards, cool down, add in NaHCO₃Aqueous solution tune PH to 6-7, EA are extracted, and organic phase is washed with water, and anhydrous sodium sulfate drying is spin-dried for molten Agent obtains crude product.Crude product chromatographic column cross column purify faint yellow color solid N- (5- (4- (3- (2,3,6- trifluoro-benzyls amino)- 3- oxopropyls amino) -5- chlorine pyrimidine radicals -2- amino) -2- fluorophenyls)-N methacrylamide (72.7mg, 39.9% Yield, PE:EA=1：1).1H NMR (DMSO-d6) δ 2.44 (t, J=6.7Hz, 2H, CH2), 3.18 (s, 3H, CH3), 3.56-3.62 (m, 2H, CH2), 4.34 (d, J=4.8Hz, 2H, CH2), 5.59 (d, J=9.6Hz, 1H, CH), 6.02-6.09 (m, 1H, CH), 6.17-6.21 (m, 1H, CH), 7.06-7.11 (m, 2H, Ar-H), 7.19 (t, J=9.3Hz, 1H, NH), 7.37-7.45(m,1H,Ar-H),7.72-7.73(m,1H,Ar-H),7.78-7.80(m,1H,Ar-H),7.94(s,1H, ), miazine-H 8.38 (m, J=4.9Hz, 1H, miazine-NH), 9.41 (s, 1H, benzene-NH) .LC-MS (m/z) 537 (M+1)。

The preparation of embodiment 9N- (3- nitrobenzophenones) acrylamide

According to the method for embodiment 2, by 3- nitroanilines (5.0g, 36.2mmol) and acryloyl chloride (6mL, 73.3mmol) Yellow solid N- (3- nitrobenzophenones) acrylamide (4.3g, 62.3%yield) is prepared.LC-MS(m/z)193(M+1).

The preparation of 10 N- of embodiment (3- aminophenyls) acrylamide

According to the method for embodiment 3, brown is prepared by N- (3- nitrobenzophenones) acrylamides (4.0g, 20.8mmol) Solid N- (3- aminophenyls) acrylamide (2.4g, 71.1%yield).LC-MS(m/z)163(M+1).

The preparation of 11 3- of embodiment (the chloro- pyrimidine radicals -4- amino of 2- (3- acrylamidos phenyl amino) -5-) propionic acid

According to method disclosed in embodiment 5, by 3- (2,5- dichloro pyrimidine base -4- amino) propionic acid (0.23g, 1mmol) and Gray solid 3- (2- (3- acrylamido phenylaminos are prepared in N- (3- aminophenyls) acrylamides (0.18g, 1.1mmol) Base) the chloro- pyrimidine radicals -4- amino of -5-) propionic acid (0.23g, 65%yield).LC-MS(m/z)362(M+1).

12 N- of embodiment (3- (the chloro- 4- of 5- (3- oxygen -3- (pyridyl group -4- methylaminos) propylcarbamic) pyrimidine radicals -2- ammonia Base) phenyl) acrylamide preparation

According to method disclosed in embodiment 6, by the 3- (chloro- pyrimidine radicals -4- ammonia of 2- (3- acrylamidos phenyl amino) -5- Base) propionic acid (36mg 0.1mmol) and 4- aminomethyl-pyridines (43mg, 0.4mmol) be prepared white solid N- ((5- be chloro- by 3- 4- (3- oxygen -3- (pyridyl group -4- methylaminos) propylcarbamic) pyrimidine radicals -2- amino) phenyl) acrylamide (26mg, 58% yield).1H NMR (DMSO-d6) δ 2.58 (t, J=6.8Hz, 2H, CH2), 3.72-3.74 (m, 2H, CH2), 4.31 (d, J= 5.6Hz, 2H, CH2), 5.66 (d, J=10.2Hz, 1H, CH), 6.22 (d, J=16.8Hz, 1H, CH), 6.45-6.50 (m, 1H, ), CH 7.14 (t, J=8.2Hz, 1H, Ar-H), 7.16-7.20 (m, 1H, NH), 7.22 (d, J=4.7Hz, 2H, Ar-H), 7.35 (d, J=8.0Hz, 2H, Ar-H), 7.95 (s, 1H, miazine-H), 8.04 (s, 1H, Ar-H), 8.46 (d, J=4.8Hz, 2H, ), Ar-H 8.64 (t, J=5.8Hz, 1H, miazine-NH), 9.27 (s, 1H, benzene-NH), 10.03 (s, 1H, benzene-NH).LC-MS(m/z)452(M+1)。

13 N- of embodiment (5- (the chloro- 4- of 5- (3- oxygen -3- (pyridyl group -4- methylaminos) propylcarbamic) pyrimidine radicals -2- ammonia Base) -2- fluorophenyls)-N methacrylamide preparation

According to method disclosed in embodiment 6, by 3- (the chloro- 2- of 5- (the fluoro- 3- of 4- (N methacrylamide base) phenyl amino) Pyrimidine radicals -4- amino) that white is prepared is solid for propionic acid (30mg, 0.08mmol) and 4- aminomethyl-pyridines (13mg, 0.12mmol) Body N- (5- (the chloro- 4- of 5- (3- oxygen -3- (pyridyl group -4- methylaminos) propylcarbamic) pyrimidine radicals -2- amino) -2- fluorophenyls) - N methacrylamide (7mg, 16.4%yield).1H NMR(DMSO-d6)δ2.63-2.65(m,2H,CH2),3.19(s, 3H, CH3), 3.51-3.56 (m, 2H, CH2), 4.39-4.41 (m, 2H, CH2), 5.62 (d, J=3.29Hz, 1H, CH), 6.08- 6.21(m,2H,CH),7.22-7.24(m,4H,Ar-H),7.71-7.72(m,1H,Ar-H),7.76-7.86(m,1H,Ar-H), 7.91-7.98 (m, 1H, Ar-H), 8.47 (s, 1H, miazine-H), 8.56-8.58 (m, 2H, miazine-NH, NH), 9.49 (s,1H,benzene-NH).LC-MS(m/z)484(M+1).

14 N- of embodiment (5- (the chloro- 4- of 5- (3- oxygen -3- (pyridyl group -4- ethyls amido) propylcarbamic) pyrimidine radicals -2- ammonia Base) -2- fluorophenyls)-N methacrylamide preparation

According to method disclosed in embodiment 6, by 3- (the chloro- 2- of 5- (the fluoro- 3- of 4- (N methacrylamide base) phenyl amino) Pyrimidine radicals -4- amino) that white is prepared is solid for propionic acid (30mg, 0.08mmol) and 4- aminoethyls pyridine (15mg, 0.12mmol) Body N- (5- (the chloro- 4- of 5- (3- oxygen -3- (pyridyl group -4- ethyls amido) propylcarbamic) pyrimidine radicals -2- amino) -2- fluorophenyls) - N methacrylamide (5mg, 12.6%yield).1H NMR (DMSO-d6) δ 2.39 (t, J=6.6Hz, 2H, CH2), 2.72 (t, J=7.0Hz, 2H, CH2), 3.18 (s, 3H, CH3), 3.28-3.34 (m, 2H, CH2), 3.55-3.60 (m, 2H, CH2), 5.60 (d, J=10.4Hz, 1H, CH), 6.05-6.09 (m, 1H, CH), 6.16-6.20 (m, 1H, CH), 7.16-7.18 (m, 1H, ), NH 7.20 (d, J=5.6Hz, 2H, Ar-H), 7.24-7.27 (m, 1H, Ar-H), 7.66-7.71 (m, 1H, Ar-H), 7.84- 7.86 (m, 1H, Ar-H), 7.94 (s, 1H, miazine-H), 7.96-7.98 (m, 1H, miazine-NH), 8.43 (d, J= 5.5Hz,2H,Ar-H),9.42(s,1H,benzene-NH).LC-MS(m/z)498(M+1)。

The preparation of 15 5- of embodiment (2,5- dichloro pyrimidine base -4- amino) valeric acid

According to method disclosed in embodiment 4, by 2,4,5- trichloropyrimidines (156mg, 0.85mmol) and 5- aminovaleric acids (100mg, 0.85mmol) be prepared white solid 5- (2,5- dichloro pyrimidine base -4- amino) valeric acid (200mg, 80.5% yield)。LC-MS(m/z)264(M+1)。

The preparation of 16 5- of embodiment (the chloro- pyrimidine radicals -4- amino of 2- (3- acrylamidos phenyl amino) -5-) valeric acid

According to method disclosed in embodiment 5, by 5- (2,5- dichloro pyrimidine base -4- amino) valeric acid (100mg, 0.38mmol) White solid 5- (2- (3- acrylamido phenyl is prepared with N- (3- aminophenyls) acrylamides (62mg, 0.38mmol) Amino) the chloro- pyrimidine radicals -4- amino of -5-) valeric acid (60mg, 40.5%yield).LC-MS(m/z)390(M+1).

17 5- of embodiment (2- (3- acrylamides phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (pyridyl group -4- first Base) pentanamide preparation

According to method disclosed in embodiment 6, by the 5- (chloro- pyrimidine radicals -4- ammonia of 2- (3- acrylamidos phenyl amino) -5- Base) white solid 5- (2- (3- third are prepared in valeric acid (30mg, 0.08mmol) and 4- aminomethyl-pyridines (12mg, 0.12mmol) Acrylamide phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (pyridyl group -4- methyl) pentanamide (19mg, 51.5%yield). 1H NMR(DMSO-d6)δ1.55-1.60(m,4H,2×CH2),2.19-2.21(m,2H,CH2),3.42-3.44(m,2H, ), CH2 4.26 (d, J=5.91Hz, 2H, CH2), 5.69-5.72 (m, 1H, CH), 6.21-6.25 (m, 1H, CH), 6.42-6.49 (m, 1H, CH), 7.09-7.28 (m, 5H, miazine-NH, Ar-H), 7.45 (d, J=7.88Hz, 1H, Ar-H), 7.91 (s, 1H, miazine-H), 8.06 (s, 1H, Ar-H), 8.39 (t, J=5.88Hz, 1H, NH), 8.45 (d, J=5.84Hz, 2H, Ar- H),9.19(s,1H,benzene-NH),10.04(s,1H,benzene-NH).LC-MS(m/z)480(M+1)。

18 5- of embodiment (2- (3- acrylamides phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (2- aminophenyls) The preparation of pentanamide

According to method disclosed in embodiment 6, by the 5- (chloro- pyrimidine radicals -4- ammonia of 2- (3- acrylamidos phenyl amino) -5- Base) white solid 5- (2- (3- acrylamides are prepared in valeric acid (39mg 0.1mmol) and o-phenylenediamine (43mg, 0.4mmol) Phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (2- aminophenyls) pentanamide (30mg, 62%yield).1H NMR (DMSO-d6)δ1.60-1.64(m,4H,2×CH2),2.31-2.35(m,2H,CH2),3.43-3.47(m,2H,CH2),4.80 (s, 2H, NH2), 5.70 (d, J=9.48Hz, 1H, CH), 6.23 (d, J=16.84Hz, 1H, CH), 6.41-6.45 (m, 1H, CH),6.49-6.51(m,1H,Ar-H),6.69-6.71(m,1H,Ar-H),6.87-6.93(m,1H,Ar-H),7.13-7.45 (m,4H,Ar-H),7.44-7.46(m,1H,miazine-NH),7.91(s,1H,miazine-H),7.99(s,1H,Ar-H), 9.07(s,1H,benzene-NH),9.19(s,1H,benzene-NH),10.01(s,1H,benzene-NH).LC-MS(m/z) 480(M+1)。

19 5- of embodiment (2- (3- acrylamides phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (2- amino -4- fluorine Phenyl) pentanamide preparation

According to method disclosed in embodiment 6, by the 5- (chloro- pyrimidine radicals -4- ammonia of 2- (3- acrylamidos phenyl amino) -5- Base) yellow solid 5- (2- (3- propylene is prepared in valeric acid (39mg 0.1mmol) and 4- fluorine o-phenylenediamine (50mg, 0.4mmol) Amide phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (2- amino -4- fluorophenyls) pentanamide (25mg, 50%yield).1H NMR(DMSO-d6)δ1.60-1.64(m,4H,2×CH2),2.31-2.35(m,2H,CH2),3.44-3.48(m,2H,CH2), 5.13 (s, 2H, NH2), 5.72 (d, J=9.76Hz, 1H, CH), 6.25 (d, J=16.64Hz, 1H, CH), 6.27-6.28 (m, 1H, Ar-H), 6.43-6.50 (m, 2H, CH, Ar-H), 7.03-7.20 (m, 4H, Ar-H), 7.46-7.48 (m, 1H, miazine- NH),7.92(s,1H,miazine-H),8.02(s,1H,Ar-H),9.03(s,1H,benzene-NH),9.21(s,1H, benzene-NH),10.04(s,1H,benzene-NH).LC-MS(m/z)498(M+1)。

20 5- of embodiment (2- (3- acrylamides phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (2- amino -4- three Trifluoromethylphenyl) pentanamide preparation

According to method disclosed in embodiment 6, by the 5- (chloro- pyrimidine radicals -4- ammonia of 2- (3- acrylamidos phenyl amino) -5- Base) black solid 5- (2- are prepared in valeric acid (39mg 0.1mmol) and 4- trifluoromethyls o-phenylenediamine (70mg, 0.4mmol) (3- acrylamides phenyl amino) -5- chlorine pyrimidine radicals -4- amino)-N- (2- amino -4- trifluoromethyls) pentanamide (32mg, 69%yield).1H NMR(DMSO-d6)δ1.60-1.64(m,4H,2×CH2),3.38-3.40(m,2H,CH2),3.45- 3.47 (m, 2H, CH2), 5.63 (s, 2H, NH2), 5.69 (d, J=10.1Hz, 1H, CH), 6.23 (d, J=16Hz, 1H, CH), 6.44-6.50 (m, 1H, CH), 6.8 (d, J=8.4Hz, 1H, Ar-H), 7.14-7.15 (m, 1H, Ar-H), 7.16 (s, 1H, Ar- ), H 7.18-7.20 (m, 1H, Ar-H), 7.20-7.22 (m, 1H, Ar-H), 7.45 (d, J=7.7Hz, 1H, Ar-H), 7.63- 7.65(m,1H,miazine-NH),7.90(s,1H,miazine-H),8.02(s,1H,Ar-H),9.18(s,1H,benzene- NH),9.23(s,1H,benzene-NH),10.06(s,1H,benzene-NH).LC-MS(m/z)548(M+1).

The preparation of 21 N- of embodiment (the fluoro- 5- nitrobenzophenones of 2-) acrylamide

According to method disclosed in embodiment 2, by the fluoro- 5- nitroanilines (0.3g, 2.0mmol) of 2- and acryloyl chloride Yellow solid N- (the fluoro- 5- nitrobenzophenones of 2-) acrylamide (0.3g, 71.4%yield) is prepared in (0.27g, 3mmol). LC-MS(m/z)211(M+1)。

The preparation of 22 N- of embodiment (5- amino -2- fluorophenyls) acrylamide

According to method disclosed in embodiment 3, made by N- (the fluoro- 5- nitrobenzophenones of 2-) acrylamide (0.25g, 4.7mmol) It is standby to obtain brown solid N- (5- amino -2- fluorophenyls) acrylamide (0.1g, 46.7%yield).LC-MS(m/z)181(M+ 1)。

The system of 23 5- of embodiment (2- (3- acrylamido -4- Fluorophenylaminos) -5- chlorine pyrimidine radicals -4- amino) valeric acid It is standby

According to method disclosed in embodiment 5, by 5- (2,5- dichloro pyrimidine base -4- amino) valeric acids (53mg, 0.2mmol) and N- (5- amino -2- fluorophenyls) acrylamide (40mg, 0.4mmol) be prepared gray solid 5- (2- (3- acrylamidos - 4- Fluorophenylaminos) -5- chlorine pyrimidine radicals -4- amino) valeric acid (40mg, 50%yield).LC-MS(m/z)408(M+1).

24 5- of embodiment (2- (3- acrylamide -4- Fluorophenylaminos) -5- chlorine pyrimidine radicals -4- amino)-N- (2- amino - 4- fluorophenyls) pentanamide preparation

According to method disclosed in embodiment 6, by 5- (2- (3- acrylamido -4- Fluorophenylaminos) -5- chlorine pyrimidine radicals - 4- amino) white solid 5- (2- (3- are prepared in valeric acid (40mg, 0.1mmol) and 4- fluorine o-phenylenediamine (50mg, 0.4mmol) Acrylamide -4- Fluorophenylaminos) -5- chlorine pyrimidine radicals -4- amino)-N- (2- amino -4- fluorophenyls) pentanamide (30mg, 60% yield)。1H NMR(DMSO-d6)δ1.49-1.61(m,4H,2×CH2),2.36-2.38(m,2H,CH2),3.36-3.38 (m, 2H, CH2), 5.26 (s, 2H, NH2), 5.73 (d, J=9.9Hz, 1H, CH), 6.26 (d, J=15.6Hz, 2H, Ar-H), 6.48 (d, J=10.6Hz, 1H, CH), 6.49-6.64 (m, 1H, CH), 7.12-7.14 (m, 1H, Ar-H), 7.15-7.16 (m, 1H,Ar-H),7.20-7.22(m,1H,Ar-H),7.48-7.49(m,1H,miazine-NH),7.94(s,1H,miazine- H),8.27(s,1H,Ar-H),9.24(s,1H,benzene-NH),9.30(s,1H,benzene-NH),9.95(s,1H, benzene-NH).LC-MS(m/z)516(M+1)。

25 the enzyme activity of embodiment detects

Test cardinal principle

The basic principle of external zymetology Activity determination is the specific substrate using fluorescent marker, is carried out under zymogenesis Phosphorylation, phosphorylated substrate and phosphorylated substrate different wave length can generate fluorescence signal of different strengths and weaknesses (445nm with 520nm).When adding in different test compounds, the degree of substrate phosphorylation is presented as to the inhibition of kinase activity, so as to table Reveal different fluorescence signals, and inhibitory activity of the compound to kinases is calculated with this.Basic testing principle as shown in Figure 1, Fig. 1 is kinase activity testing principle schematic diagram of the embodiment of the present invention.

The detection of zymetology inhibitory activity using the people of GST marks recombinate JAKs kinases JAK1/PV4774, JAK2/PV4210, JAK3/PV3855, TYK2/PV4790 and its corresponding specific substrate Tyr6 (Kinase Assay Kit- Tyrosine6Peptide, JAK1/PV4122), Tyr4 (Kinase Assay Kit- Tyrosine4Peptide, JAK2/PV3193), Tyr4 (Kinase Assay Kit- Tyrosine4Peptide, JAK3/PV3193), Tyr3 (Kinase Assay Kit- Tyrosine4Peptide, TYK2/PV3192).Detection reagent be color developing agent A (Development reagent A, PV3297).Above-mentioned all material is bought from Invitrogen companies.

Test main process

The flow that experimentation is required according to detection reagent operation instructions carries out (Invitrogen).Flow is as follows：

(1) Preparatory work of experiment：Kinase reaction buffer solution (working solution) is prepared as requested, will be tested with kinase reaction buffer solution Diluted chemical compound (detects JAK1, JAK2, compound maximum concentration is 10 μM into various concentration gradient；JAK3 is detected, is changed It is 1 μM to close object maximum concentration).

(2) 10 μ L of zymetology reaction system, including 2.5 μ L tests compound, 5 μ L kinase reactions buffer solutions and 2.5 μ L ATP Solution (kit offer) after mixing, reacts at room temperature 1h.

(3) detection reaction is provided with control reaction simultaneously, and 0 including being not added with test compound inhibits control, is not added with The 0 phosphorylation control of ATP and the positive control of addition phosphorylated substrate.All detections are using multiple holes.

(4) zymetology after reaction, adds in the chromogenic reaction buffer solution that 5 μ L are prepared in advance, reacts at room temperature 1h.Then add in 5 μ L stop buffers terminate reaction.

(5) detected using fluorescence detector (Ascent Fluoroskan FL reader, Thermo Labsystems) Fluorescence signal per hole, excitation wavelength 400nm, the launch wavelength of detection is respectively 445nm and 520nm.Substrate phosphorylation Ratio is with reference to fluorescence signal intensity C445/F520.

(6) the zymetology inhibiting rate calculation formula of compound is tested：Inhibiting rate (%)=1- detection holes substrate phosphorylation/0 presses down Control wells phosphoric acid rate processed.

The positive control of selection is CP690550, and structural formula is as follows：

According to above-mentioned experimental method, external zymetology Activity determination, experimental result are carried out to compound of Formula I of the present invention 2 are shown in Table, table 2 provides compound to the external inhibitory activity result of jak kinase for the embodiment of the present invention.

2 compound of Formula I of the present invention of table is to the external inhibitory activity result of jak kinase

Embodiment	JAK3	JAK2	JAK1
				CP-690550	1	1	1
6	0.88	0.03	0.50
				8	0.88	0	0.11
12	1.06	0	0.11
				13	0.69	0	0.26
14	1.03	0.04	0.27
				17	0.70	0	0.24
18	1.08	0	0.88
				19	1.22	0.09	0.87
20	0.65	0	0.48
				24	1.12	0.07	0.7

In table 1, the inhibitory activity of embodiment compound is the inhibitory activity compared with positive control.

In table 1, to JAK3, JAK2, test compound concentration is 30nM；JAK1 is detected, test compound concentration is 300nM。

As shown in Table 1, compared with full inhibitor C P-690550, compound provided by the invention has selection to jak kinase Property inhibitory action, it is more notable to JAK3 kinase inhibitory activities, and it is not notable to JAK2 kinases and JAK1 kinase inhibiting activities, it can For use as selective JAK 3 kinases inhibitor.

The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications also should It is considered as protection scope of the present invention.

Claims

1. pyrimidine derivatives or its pharmaceutically acceptable salt shown in formula (I)：

Wherein,

A is phenyl ring or pyridine ring；

X is halogen；

R¹Independently selected from hydrogen, cyano, halogen, amino, hydroxyl, the alkyl of C1~C6 or halogenated alkyl, C1~C6 alkoxy in One or more；

R²Independently selected from one or more of hydrogen or halogen；

R³Selected from hydrogen or methyl；

N is 1 to 7 integer；

P is 0 to 3 integer.

2. pyrimidine derivatives according to claim 1, which is characterized in that R¹Independently selected from hydrogen, cyano, fluorine, amino, three One or more of methyl fluoride, R²Selected from hydrogen or fluorine, R³Selected from hydrogen or methyl.

3. pyrimidine derivatives according to claim 1, which is characterized in that A is phenyl ring；R¹Independently selected from fluorine, amino, three One or more of methyl fluoride；R²For fluorine；R³For methyl；N is 4, p 0.

4. pyrimidine derivatives according to claim 1, which is characterized in that there is formula (1)~formula (10) structure：

5. a kind of preparation method of the pyrimidine derivatives shown in formula (I), including：

Logical formula (VI) compound and logical formula (VII) compound are formed into general formula as (I) under organic solvent and catalyst action Compound；

Wherein,

A is phenyl ring or pyridine ring；

R²Independently selected from one or more of hydrogen or halogen；

R³Selected from hydrogen or methyl；

N is 1 to 7 integer；

P is 0 to 3 integer.

6. preparation method according to claim 5, which is characterized in that the catalyst is selected from 1- ethyls -3- (3- dimethylamine Propyl) carbodiimide, N, N '-dicyclohexylcarbodiimide or N, N '-phosphinylidyne diimidazole.

7. preparation method according to claim 5, which is characterized in that the organic solvent is selected from benzene, toluene, tetrahydrochysene furan It mutters, Isosorbide-5-Nitrae-dioxane, dichloromethane, chloroform or n,N-Dimethylformamide.

8. preparation method according to claim 5, which is characterized in that the logical formula (VI) compound is made in accordance with the following methods It is standby：

Logical formula (IV) compound and logical formula (V) compound are formed into general formula as (VI) under the action of organic solvent and catalyst Compound；

9. preparation method according to claim 8, which is characterized in that the catalyst is selected from trifluoroacetic acid, hydrochloric acid or first Sulfonic acid.

10. preparation method according to claim 8, which is characterized in that the organic solvent is selected from isopropanol or n-butanol.

11. a kind of pharmaceutical preparation, including the pyrimidine derivatives described in Claims 1 to 4 any one or its is pharmaceutically acceptable Salt and pharmaceutically acceptable auxiliary material.

12. pharmaceutical preparation according to claim 11, which is characterized in that the preparation is tablet, capsule, pulvis, sugar Slurry, liquor, suspending agent, injection or paste.

13. pyrimidine derivatives or its pharmaceutically acceptable salt or claim 11 described in Claims 1 to 4 any one or Application of the pharmaceutical preparation in JAK3 kinase inhibitors are prepared described in 12.

14. pyrimidine derivatives or its pharmaceutically acceptable salt or claim 11 described in Claims 1 to 4 any one or Application of the pharmaceutical preparation in the drug of prevention or treatment with the relevant disease of JAK3 abnormal kinases is prepared described in 12.

15. application according to claim 14, which is characterized in that the disease is autoimmune disease, inflammatory disease Disease, cancer, myeloproliferative disease, bone-resorbing disease or organ-graft refection's disease.

16. application according to claim 15, which is characterized in that the disease for rheumatoid arthritis, Crohn disease, Systemic loupus erythematosus, multiple sclerosis, Type I diabetes, psoriasis, anaphylactia, asthma, chronic obstructive pulmonary disease, Leukaemia or lymthoma.