CN106397432B - A kind of compound as JAK inhibitor - Google Patents

A kind of compound as JAK inhibitor Download PDF

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CN106397432B
CN106397432B CN201510481323.1A CN201510481323A CN106397432B CN 106397432 B CN106397432 B CN 106397432B CN 201510481323 A CN201510481323 A CN 201510481323A CN 106397432 B CN106397432 B CN 106397432B
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pyrimidine
fluoro
amino
pyrazoles
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CN106397432A (en
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陈亦林
彭红
钱进
陶琳
赵岩
张晓丽
赵银鹰
邹阳
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Nanchang Hongyi Technology Co Ltd
Nanchang Hongyi Pharmaceutical Co Ltd
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Nanchang Hongyi Technology Co Ltd
Nanchang Hongyi Pharmaceutical Co Ltd
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4168—1,3-Diazoles having a nitrogen attached in position 2, e.g. clonidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/08—Solutions
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
    • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
    • C07D233/66—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
    • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
    • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
    • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
    • C07D239/46—Two or more oxygen, sulphur or nitrogen atoms
    • C07D239/48—Two nitrogen atoms

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  • Organic Chemistry (AREA)
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  • Pharmacology & Pharmacy (AREA)
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  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The present invention relates to a kind of noval chemical compound as JAK inhibitor, the compound is compound of formula I and its pharmaceutically acceptable salt, prodrug, metabolin, isotope derivatives and solvate, and the pharmaceutical composition of the compound is included, the disease and illness related to Janus kinases (JAK) in human patient, mammalian subject available for prevention or treatment；And its can as Janus kinases (JAK) inhibitor medical science, pharmacy, biology, physiology, biochemical etc. experiment in application.

Description

A kind of compound as JAK inhibitor
Technical field
The present invention relates to a kind of noval chemical compound as JAK inhibitor, including its pharmaceutically acceptable salt, prodrug, generation Thing, isotope derivatives and solvate are thanked, it can be used for adjusting cytoactive such as signal by regulatory protein kinase activity Transduction, propagation and cytokine secretion.In addition, the present invention relates to the pharmaceutical composition for including the compound, available for preventing Or the JAK inhibitor for the treatment of Janus kinases (JAK) relevant disease, and can be applied as Janus kinases (JAK) inhibitor In the experiment such as medical science, pharmacy, biology, physiology, biochemical.The JAK relevant diseases include inflammatory disease, LADA Disease, proliferative diseases, proliferative disease etc..
Background technology
Protein kinase (PK) is one group of enzyme for regulating and controlling a variety of important biomolecule processes, and the bioprocess especially swashs including cell Enzymatic protein, lipid, sugar, the phosphorylation of nucleosides and other cell metabolites and at all aspects of eukaryotic cell physiology Play a crucial role.Especially, protein kinase and lipid kinase participate in signal transduction event, and the event control is to extracellular instrumentality Or activation, growth, differentiation and the survival of the cell of stimulant (such as growth factor, cell factor or chemotactic factor (CF)) response.
Janus kinases（Janus kinase, JAK) it is a kind of non-transmembrane non-receptor type protein tyrosine kinase family, Played an important role in cytokine signaling transmittance process.Jak kinase can phosphorylation cytokine receptor combined with it, Multiple homologous 2 domains containing specific Src of and can phosphorylation（Src homology 2 domain, SH2) signaling molecule.At present There are mammal JAK family members known to four kinds：JAK1, JAK2, JAK3 and TYK2.They have 7 JAK same in structure Homeodomain（JAK homology domain, JH）, wherein JH1 domains are kinases area, and function is encoded kinases albumen； JH2 domains are "false" kinases areas, and JH1 activity is played regulatory role；JH3-JH7 forms a four-in-one domain, regulation JAK and acceptor combination.JAK3 is distributed in marrow and lymphatic system, and JAK1, JAK2, TYK2 are distributed widely in Various Tissues In cell.Jak kinase participates in many important biological processes such as propagation, differentiation, apoptosis and the immunological regulation of cell.
Signal transducer and transcription activator（signal transducer and activator of Transcription, STAT) be JAK substrate.Signal transduction and transcriptional activation（STATs) protein family includes STAT1, 7 members such as STAT2, STAT3, STAT4, STAT5a, STAT5b and STAT6.Interaction between JAKs and STATs is thin Played an important role in intracellular cytokine receptor signaling pathways（O'SULLIVAN LA,LIONGUE C,LEWIS RS,et al.Cytokine receptor signaling through the Jak Stat pathway in disease[J] .MolImmunol,2007,44{10)：2497-2506.).After cell factor is combined with the specific receptor on its target cell JAK can be activated, then phosphorylation occurs for the tyrosine residue on catalytic receptor, and forms corresponding STAT and receptor complex With reference to " docking site "（docking site).Last jak kinase catalysis stat protein phosphorylation, the STAT of activation are formed together Combined in source or the laggard people's cell core of heterodimer with specific target gene, regulation and control destination protein expression（LVASHKIV LB, HU XY.Signaling by STATs[J].Arthritis Res Ther,2004,6(4):159-168.）, this approach is JAK/STAT signal paths.The generation of a variety of diseases such as the abnormal activation of JAK/STAT signal transduction pathways and tumour, leukaemia, Development is closely related with prognosis.
JAK/STAT signal paths be in recent years newfound one turn with cell factor closely related Intracellular signals Guiding path, many important physiological processes such as propagation, differentiation, apoptosis and the immunological regulation of cell are participated in, to immunity of organism Response, immune cell differentiation development and inflammatory reaction etc. have a major impact, in diseases such as tumour, inflammation and various autoimmunes Occur, played an important role in development.The abnormal activation of JAK/STAT signal paths and kinds of tumors occurrence and development are closely related.
JAK/STAT signal paths are a signal transduction pathways by cytokine profiles receptor for stimulating, these factor bags Include interleukin class (such as IL-2~7, IL-9, IL-10, IL-15, IL-21 etc.）, interferons (including IFN-α, IFN-β, IFN- γ etc.）, hematopoietin (EPO), granulocyte and macrophage colony stimulatory factor（GM-CSF), somatotropin（GH)、 Prolactin（PRL), thrombopoietin（TPO), platelet derived growth factor（) and epithelical cell growth factor PDGF（EGF) Deng it plays a crucial role in immunological regulation, the biological process such as immune cell propagation is participated in（GHORESCHI K, LAURENCE A,O'SHEA JJ.Janus kinases in immune cell signaling [J].Immunol Rev,2009,228 (1)：273-287.）.Isoacceptor can not activate the jak kinase of different subtype, so as to show the biological function of differentiation.
JAK1 gene knockout experiments on mouse model show that the enzyme is adjusting the life of above-mentioned cytokine profiles acceptor Key effect is played in thing effect（KISSELEVA T, BHATTACHARYA S, BRAUNSTEIN J, et al.Signaling through the JAK/STAT pathway,recent advances and future challenges[J].Gene,2002,285 (1-2):1-24.）.
JAK2 is knocked out in mouse model can cause animal dead caused by anaemia（SCHINDLER C, LEVY DE, DECKER T.JAK-STAT signaling：from interferons to cytokines[J].J Biol Chem, 2007,282(28):20059-20063.）.A base mutation JAK2V617F on JAK2 genes in human body, itself and marrow Polycythemia vera in proliferative disease（PV), essential thrombocythemia（ET), idiopathic myelofibrosis （IMF), chronic myelocytic leukemia（) etc. CML generation is closely related（GHORESCHI K, LAURENCE A, O'SHEA JJ.Janus kinases in immune cell signaling [J].Immunol Rev,2009,228(1)：273- 287.）.JAK2 inhibitor has described to be applied to bone marrow proliferative diseases (Santos et al., Blood, 2010,115:1131 ； Barosi G. and Rosti V., Curr.Opin.Hematol., 2009,16:129 ；Atallah E. and Versotvsek S., 2009Exp.Rev.Anticancer Ther.9:663).
JAK3 defects are identified in the people with autosomal recessive severe combined immunodeficient (SCID) first (Macchi etc., 1995.Nature377 (6544)：65-68).JAK3 knock-out mices display that SCID but not shown nonimmune Property defect, show that JAK3 inhibitor will have correlation effect in vivo and therefore as being used for immunosupress as immunodepressant Promising medicine (Papageorgiou and Wikman2004, Trends in Pharmacological Sciences25 (11) ：558-62).The inhibitor of Tyrosine kinase JAK3 be described be suitable for immunodepressant (such as United States Patent (USP) 6, 313,129；Borie et al., Curr.Opin.Investigational Drugs, 2003,4:1297).
TYK2 is the 1st member in JAK families, and it can be by IF-Ns, 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 acceptor that defect occurs, and enter And cause virus infection, antibacterial immunity function reduction and add possibility of pulmonary infection etc.（KISSELEVA T, BHATTACHARYA S,BRAUNSTEIN J,et al.Signaling through the JAK/STAT pathway, recent advances and future challenges[J].Gene,2002,285 (1-2):1-24.）.In addition, The research of Lamer AC groups shows that TYK2 can help to suppress the growth and transfer of breast cancer（ZHANG Q,STURGILL JL, KMIECIAK M et al.The role of Tyk2 in regulation of breast cancer growth[J].J Intetferon Cytokine Res,2011,31(9)：671-677.）
In summary, disabling signal transduction is expected to exploitation and treats or prevents Janus kinases in the level of JAK kinases (JAK) relevant disease, such as immune, inflammation, autoimmunity, proliferative diseases such as cancer, proliferative disease, allergic condition or disease Sick, graft rejection or graft versus host disease(GVH disease), xerophthalmia etc..
Pfizer at present（Pfizer）The JAK inhibitor Tofacitinib energy selective depression JAK3 of company's research and development swashs Enzyme, ratified on November 6th, 2012 by FDA for treat the activities of adults phase and to methotrexate (MTX) reaction it is bad in severe class Rheumatic arthritis（RA）.Tofacitinib major side effects have severe infections rate and low-density lipoprotein white level to improve, Most common adverse reaction is the infection of the upper respiratory tract, headache, diarrhoea, nasal congestion, sore-throat and nasopharyngitis.Except fatty degeneration of liver, Outside peripheral edema, Tofacitinib other most of adverse reactions, monoclonal antibody class medicine is also all present.Tofacitinib conducts Immunodepressant, ratify specification in warning and points for attention and anti-TNF monoclonal antibody medicines it is essentially identical.Because part suppresses Jak2 activity simultaneously disturbs the cell factors such as erythropoietin(EPO) and colony stimulating factor to play effect, thus also has clinical research Report, Tofacitinib can cause the side effects such as anaemia and neutrophilic granulocytopenia.In addition, clinical test is shown, Tofacitinib can't cause T total number of lymphocytes to reduce, but can cause CD8+T Leukopenias and NK（NK Cell）Some uncertain risks also be present when slightly reducing, therefore taking Tofacitinib.[it is used to treat rheumatoid pass The scorching JAK inhibitor of section, Xue Feng, Liu Fei, Wu Gang, You Qidong,《Pharmacy is in progress》2014,38（4）：264-273]
Although a series of jak kinase inhibitor has been disclosed at present, these have listed or have been in conceptual phase Jak kinase inhibitor also has improved space in terms of efficacy and saferry, it is still necessary to develops the new of more preferable drug effect and security Compound.The compound of the present invention shows good activity and security as Janus kinases (JAK) inhibitor.
The content of the invention
The present invention relates to a kind of noval chemical compound as JAK inhibitor, the compound is formula（I）Compound and its pharmaceutically Acceptable salt, prodrug, metabolin, isotope derivatives and solvate, and the pharmaceutical composition of the compound is included, can For preventing or treating disease and illness related to Janus kinases (JAK) in human patient, mammalian subject；And its It can be applied as Janus kinases (JAK) inhibitor in the experiment such as medical science, pharmacy, biology, physiology, biochemical.
A kind of formula（I）Noval chemical compound：
（I）
Including its pharmaceutically acceptable salt, prodrug, metabolin, isotope derivatives and solvate, wherein：
Ring A is C3-7Cycloalkyl, C5-7Fragrant ring group, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11Fragrance is miscellaneous bicyclic Base, C11-15Ternary ring group, its middle ring A is optionally by one or more identical or different R, R1Substituted；
R、R1It is H, halogen, benzene, C3-7Cycloalkyl, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11Fragrance is miscellaneous bicyclic Base, optionally by one or more identical or different R wherein on these rings5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, its Middle C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl is optionally by one or more identical or different R6Substitution；
B1It is H, CH3、CN、NO2、CF3, halogen；
B2It is H, CH3、CN、NO2、CF3, halogen；
X is H, CH3、CN、NO2、CF3、C(O)NH2, halogen；
R2It is H, CH3、CN、NO2、CF3, halogen；
R3It is H, CH3、CN、NO2、CF3, halogen；
R4It is H, CN, NO2、CF3、COOH、COOR7、CONR8R8’、SONR9R9’、COR10、R11OH, halogen；Benzene, C3-7Cycloalkanes Base, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, it is wherein optionally one or more on these rings Identical or different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl optionally quilt One or more identical or different R6Substitution；
R5It is H, CN, NO2、CF3、COOH、COOR7、CONR8R8’、SONR9R9’、COR10、R11OH, halogen；Benzene, C3-7Cycloalkanes Base, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, it is wherein optionally one or more on these rings Identical or different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl optionally quilt One or more identical or different R6Substitution；
R6It is H, CN, NO2、CF3、COOH、COOR7、CONR8R8’、SONR9R9’、COR10、R11OH, halogen；Benzene, C3-7Cycloalkanes Base, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, it is wherein optionally one or more on these rings Identical or different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl optionally quilt One or more identical or different R6Substitution；
R7It is benzene, C3-7Cycloalkyl, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, wherein this Optionally by one or more identical or different R on a little rings5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl is optionally by one or more identical or different R6Substitution；
R8、R8’It is H, CN, NO respectively2、CF3、COOR7、CONR8R8’、COR10、R11OH, halogen；Benzene, C3-7Cycloalkyl, C5-7 Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, wherein on these rings optionally by one or more identical or Different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl optionally by one or Multiple identical or different R6Substitution；
R9、R9’It is H, CN, NO respectively2、CF3、COOH、COOR7、CONR8R8’、COR10、R11OH, halogen；Benzene, C3-7Cycloalkanes Base, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, it is wherein optionally one or more on these rings Identical or different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl optionally quilt One or more identical or different R6Substitution；
R10It is H, CN, NO2、CF3、COOH、COOR7、CONR8R8’、SONR9R9’、COR10、R11OH, halogen；Benzene, C3-7Cycloalkanes Base, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, it is wherein optionally one or more on these rings Identical or different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl optionally quilt One or more identical or different R6Substitution；
R11It is H, benzene, C3-7Cycloalkyl, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, wherein Optionally by one or more identical or different R on these rings5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkane Base, C2-8Alkenyl, C2-8Alkynyl is optionally by one or more identical or different R6Substitution；
Y is（CR12R13）n；
N is 0 or 1；
R12、R13It is R5；
Z1、Z2C can be respectively selected from（R14）Or N（R14）；
R14It is H, CN, NO2、CF3、COOH、COOR7、CONR8R8’、SONR9R9’、COR10、R11OH, halogen；Benzene, C3-7Cycloalkanes Base, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, it is wherein optionally one or more on these rings Identical or different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl optionally quilt One or more identical or different R6Substitution.
In the implication of the present invention, term is used as described below：
" halogen " refers to F, Cl, Br, I, At.
“C3-7Cycloalkyl " refers to the cycloalkyl chain with 3-7 carbon atom, such as cyclopropyl, cyclobutyl, cyclopenta, ring Hexyl, cyclohexenyl group, suberyl.The substituent that each hydrogen of cycloalkyl carbon can be further provided for is replaced.
“C5-7Fragrant heterocyclic radical " refers to the fragrant heterocyclic radical with 5-7 carbon atom, such as imidazoles, thiazole, pyrrole Azoles, pyridine, pyrimidine etc..The substituent that each hydrogen of fragrant heterocyclic radical can be further provided for is replaced.
“C7-11Aromatic bicyclic base " refers to the aromatic bicyclic base with 7-11 carbon atom, such as naphthalene, indenes etc..Fragrance The substituent that each hydrogen of bicyclic group can be further provided for is replaced.
“C7-11The miscellaneous bicyclic group of fragrance " refers to the fragrant miscellaneous bicyclic group with 7-11 carbon atom, such as quinoline, different Quinoline, benzothiazole etc..The substituent that each hydrogen of the miscellaneous bicyclic group of fragrance can be further provided for is replaced.
“Cl-8Alkyl " refers to the alkyl chain with 1-8 carbon atom, such as：Methyl, ethyl, n-propyl, isopropyl, just Butyl, isobutyl group, sec-butyl, the tert-butyl group.Cl-8The substituent that each hydrogen of alkyl carbon can be further provided for is replaced.
“C2-8Alkenyl " refers to the alkenylene chain with 2-8 carbon atom, such as：- CH=CH, one CH=CH-CH3,-CH2-CH= CH2,-CH=CH-CH2-CH3,-CH=CH-CH=CH2。C2-8The substituent that each hydrogen of alkenyl carbon can be further provided for is replaced.
“C2-8Alkynyl " refers to the alkynyl chain with 2-8 carbon atom, such as：- C-CH ,-CH. ,-C-CH, CH2- CH2-C tri- CH, CH2-C-C-CH3.C2-6The substituent that each hydrogen of alkynyl carbon can be further provided for is replaced.
Tofacitinib：
Decernotinib：
Filgotinib：
For formula (I) compound, present invention additionally comprises all dynamic isomers and stereoisomer form of all proportions And its mixture, and its pharmaceutically acceptable salt, prodrug, metabolin, isotope derivatives and solvate, and comprising described The pharmaceutical composition of compound.
The pharmaceutically acceptable salt of formula (I) compound, comprising one or more alkalescence or acidic-groups, present invention additionally comprises Its corresponding acceptable salt pharmaceutically or in toxicology, particularly its pharmaceutically available salt.Therefore, comprising acidic-group Formula (I) compound can be used according to the invention, such as alkali metal salt, alkali salt or as ammonium salt.Such salt More accurate example include sodium salt, sylvite, calcium salt, magnesium salts or with ammonia or organic amine such as ethamine, monoethanolamine, triethanolamine or ammonia The salt of base acid.May be present and can according to the present invention in the form of it is with the addition salts of inorganic acid or organic acid use comprising one or Multiple basic groups, formula (I) compound for the group that can be protonated.The example of appropriate acid include hydrochloric acid, sulfuric acid, phosphoric acid, Nitric acid, methanesulfonic acid, lactic acid, malic acid, maleic acid, benzoic acid, tartaric acid, oxalic acid, p-methyl benzenesulfonic acid etc. and art technology Other known acid of personnel.If formula (I) compound in intramolecular simultaneously comprising acid and basic group, present invention additionally comprises except Inner salt or betaine outside the salt form referred to（Amphion）.Each salt of formula (I) can be by those skilled in the art The conventional method known obtains, such as by making these contact acquisition in solvent or dispersant with organic or inorganic acid or alkali, or By carrying out anion exchange or cation exchange acquisition with other salt.Present invention additionally comprises all salt of formula (I) compound, its Because low physiological compatible is not directly applied for medicine, but it can be used for example as the intermediate of chemical reaction or for preparing Pharmaceutically acceptable salt.
In the present invention, term " pharmaceutically acceptable " refers to that corresponding compound, carrier or molecule are suitable for administration to people. Preferably, the term refers to by management organization such as CFDA（China）、EMEA（Europe）, any national management such as FDA (U.S.) Agency qualification is used for the preferred people of mammal.
" prodrug " refer to by with enzyme, hydrochloric acid in gastric juice etc. in physiological conditions in vivo for example by each enzymatically entering The reactions such as capable oxidation, reduction, hydrolysis are converted into the derivative of the compounds of this invention.
" metabolin " refers to all molecules that any compound of the present invention is derived from cell or the preferred people of organism.
" isotope derivatives " refer to that sentencing unnatural proportions with one or more atoms of composition compound contains isotope Described compound.Such as deuterium (2 H or D), carbon -13 (13 C), nitrogen -15 (15 N) etc..
" solvate " refers to the compound form generally combined by solvolysis reaction with solvent physical.This physics knot Conjunction includes Hydrogenbond.Conventional solvents include water, ethanol, methanol, acetic acid etc..Formula（I）Compound can prepare in crystalline form and It can be in solvate form thereof（Such as hydrated form）.Suitable solvent compound includes pharmaceutically acceptable solvate and (such as is hydrated Thing), and further include stoichiometric solvates and non-stoichiometric solvates.In some cases, such as when one Or multiple solvent molecules, when including in the lattice of crystalline solid, solvate can dissociate." solvate " covers solution And solvate can be dissociated.Representative solvents compound includes hydrate, ethanolates and methanol solvate etc..
Formula (I) compound can exist with crystal or amorphous form.In addition, some crystal forms of formula (I) compound can Exist with polymorphic forms, it is included in the scope of the present invention.X- can be included but is not limited to using many conventional analytical techniques Ray powder diffraction (XRPD) figure, infrared (IR) spectrum, Raman spectrum, differential scanning calorimetry (DSC), thermogravimetric analysis (TGA) Characterized with solid-state nuclear magnetic resonance (ssNMR) to distinguish the polymorphic of formula (I) compound.
" pharmaceutical composition " when as medicine, the salt of formula I and compound of formula I, isotope derivatives, Metabolin, prodrug, solvate and the composition with bioactivity and/or without bioactive substance composition press down as JAK Preparation is treating or preventing immune, LADA or allergic conditions, proliferative disease or proliferative diseases, inflammation, allergic disease Disease, graft rejection, it is immune-mediated in application.
The pharmaceutical composition of the present invention can contain one or more pharmaceutically acceptable carriers, can be used as that injection is made With the pharmaceutical preparation and pharmaceutical dosage form of Non-parenteral Delivery Routes.The carrier include pharmaceutical field it is all can be used for injection is made With the pharmaceutical preparation of Non-parenteral Delivery Routes, such as diluent, wetting agent, filler, adhesive, wet and slippery dose, disintegrant, absorption Accelerator, surfactant, retarding agent, adsorbent, suspending agent, flocculant, deflocculant, emulsifying agent, conventional matrix, solubilising Agent, cosolvent, cosolvent, preservative, flavouring, colouring agent, antioxidant, buffer, bacteriostatic agent, isotonic regulator, PH regulations Agent, complexing of metal ion agent, curing agent, thickener, sorbefacient etc..
Formula (I) compound and pharmaceutical composition of the present invention can be made into the pharmaceutical preparation and medicine of injection or Non-parenteral Delivery Routes Agent type.Suitable for hypodermic injection, intramuscular injection, intravenous injection, oral, lung（Nose or oral cavity suction）, rectum, part, stomach Outside, intra-articular, eye, nasal-cavity administration etc., although optimal approach is by dependent on the disease to be treated in the case of any given The property and the order of severity and active component property of diseased state.They easily can be present in single formulation, and by It is prepared by any means known to pharmaceutical field.
The related disease of Janus kinases (JAK) and illness are immune, inflammation, autoimmunity, proliferative diseases in the present invention Such as cancer, proliferative disease, allergic condition or disease, graft rejection or graft versus host disease(GVH disease), xerophthalmia.
Autoimmune disease is at least partly by the immune anti-of body-defence itself component such as protein, lipid or DNA The disease that should trigger.The example of organ specific autoimmune disease disease is the insulin-dependent diabetes mellitus (I for influenceing pancreas Type), influence thyroid Hashimoto thyroiditis and Graves disease, influence the pernicious anaemia of stomach, influence adrenal Cushing disease With Addison's disease, the CAH of influence liver；Stein-Leventhal syndrome (PCOS), chylous diarrhea, psoriasis, inflammatory bowel Sick (IBD) and ankylosing spondylitis.The example of non-organ specific autoimmune disease disease is rheumatoid arthritis, multiple Hardening, systemic loupus erythematosus and myasthenia gravis.
IBD (IBD) is one group of immune-mediated chronic nonspecific bowl inflammatory diseases, main to include bursting Ulcer colitis（) and Crohn disease UC（), CD it is one of important kind of gastrointestinal tract inflammation disease.Crohn disease most often relates to And terminal ileum and colon, and be transmural and discontinuous.On the contrary, in ulcerative colitis, inflammation be it is continuous simultaneously And it is limited to rectum and colon mucosa.Limit to ileum and colon about 10% in the case of, Crohn disease or exedens knot The determination classification of enteritis can not be made, and be referred to as " uncertain colitis ".Two kinds of diseases all include skin, eyes or The parenteral inflammation in joint.The injury of neutrophil leucocyte induction can be prevented by using neutrophil migration inhibitor (Asakura etc., 2007, World J Gastroenterol.13 (15)：2145-9).
Systemic loupus erythematosus (SLE) is the chronic inflammatory disease as caused by B- cell-stimulatings cell-mediated T-, and it is led Cause glomerulonephritis and kidney failure.Features of the people SLE in early stage is the expansion of lasting autoreactivity CD4+ memory cells (D'Cruzetal.,2007, Lancet 369 (9561):587-596)。
Rheumatoid arthritis（RA）It is a kind of chronic, systemic itself to be exempted from as what is mainly showed using symmetry, panarthritis Epidemic disease, lesion mainly involve joint synovial joint and it is abarticular performance it is extensive and changeable, ultimately result in articulation structure and break It is higher that bad, function loses disability rate.
Multiple sclerosis（Multiple sclerosis, MS) it is central nervous system white matter inflammatory demyelinating itself Immunity disease, myelinoclasis and the lymphocyte of infiltration（CD4+ T) cellular immunity of mediation is relevant.SOCS1 can suppress JAK2 The STAT3 of induction phosphorylation, suppression of the SOCS1 to JAK2-STAT3 can be simulated using JAK2 inhibitor (AG490) and made With.（SOCS1-JAK2-STAT3 signal paths act on machine in C57BL/6 mice with experimental autoimmune encephalomyelitis models System is inquired into, Dong Mei etc., China Immunology Journal, and 2014,30（4）：459~463）.
Type i diabetes are secondary by selective attack of the autoreactive T cell to the beta Cell of islet of excreting insulin. It is based on the observation that in this disease by target of JAK3：The known various kinds of cell by JAK approach conducted signals because Son participates in the autoimmune injury of the T cell mediation of β cells.In fact, JAK3 inhibitor, JANEX-1 is in type i diabetes NOD mouse models in show the development for preventing spontaneous autoimmune diabetes.
It is used to treat or prevent another aspect of the invention is the compound of the present invention or its pharmaceutically acceptable salt and increases The method of raw disease especially cancer.Cancer includes the disease of one group of uncontrolled growth for being characterised by abnormal cell and diffusion.It is logical Often, cancer classification is cancer (e.g., prostate cancer, kidney, liver cancer, cancer of pancreas, stomach cancer, colorectal cancer, the mammary gland of solid tumor The graceful disease of cancer, cervical carcinoma, lung cancer, incidence cancer, thyroid cancer, glioblastoma, Kaposi's sarcoma, Karst Lay, black Plain knurl etc.), (e.g., lymthoma, leukaemia, such as acute lymphoblastic leukemia, acute myelocytic are white for hematologic cancers Blood disease (AML) or Huppert's disease), cutaneum carcinoma（Such as skin T cell lymphoma (CTCL) and skin B cell lymphomas And exemplary skin T cell）, lymthoma（Including Sai Zeli syndromes (Sezary syndrome) and mycosis fungoides） Deng.
Graft rejection（Allograft rejection）Including but not limited to such as kidney, heart, liver, lung, marrow, skin and angle Acute and chronic allo-rojection after the transplanting of film.Specific immune response of the known T cell in allo-rojection In play a crucial role.Super acute, acute and chronic organ-graft refection can treat.A few minutes in transplanting occurs for hyperacute rejection In clock.Acute cellular rejection generally occurs within six to 12 months of transplanting.Super acute and acute cellular rejection be typically it is reversible, its In use immunosuppressant treatment.The chronic rejection for being characterized as the gradual loss of organ dysfunction is that transplanting recipient is persistently concerned about, Because its can be after the transfer any time occur.
Graft versus host disease(GVH disease) (GVDH) is allogenic bone marrow transplantation (BMT) major complications.GVDH is by identification tissue phase Recipient's difference in capacitive complication system and the donor T-cells that it is reacted are caused, which results in significant morbidity Rate and the death rate.JAK3 plays a crucial role in GVHD is induced, and carrying out treatment with JAK3 signatures preparation JANEX-1 shows weakening GVHD seriousness (summarize Cetkovic-Cvrlje and Ucken, 2004).
Xerophthalmia (DES, also referred to as keratoconjunctivitis sicca) is one of most common problem of oculist's treatment.Have When DES be referred to as infull syndrome (Jackson, 2009.Canadian the Journal Ophthalmology44 of Tear function (4), 385-394).DES influences to be up to population of 10% age between 20 to 45 years old, and percentage increases with the age.To the greatest extent Pipe can utilize the artificial tear products of numerous species, but these products only provide the respite of symptom.Therefore, it is necessary to treat dry Preparation, composition and the treatment method of eye.Dry eyes are otherwise referred to as keratoconjunctivitis sicca, and the treatment of xerophthalmia includes improving The specific symptoms of xerophthalmia, such as ophthalmic uncomfortable, dysopia, tear film be unstable, the inflammation of tears hyperosmosis and eyeball surface.
Therefore, another aspect of the invention is the compound of the present invention and its pharmaceutically acceptable salt, prodrug, metabolism Thing, isotope derivatives and solvate, and the pharmaceutical composition of the compound is included, it is immune, scorching for preventing or treating Disease, autoimmunity, proliferative diseases such as cancer, proliferative disease, allergic condition or disease, graft rejection or graft-versus-host The method of disease, xerophthalmia etc..
Beneficial effect of the present invention
Pass through the implementation of technical solution of the present invention, the results showed that compound of the present invention（Formulas I）With good suppression JAK activity processed and pharmacokinetic property, have biological similarities, no cytotoxicity, rat single oral with Tofacitinib Gastric infusion has no overt toxicity.
Embodiment
Embodiment 1：2- (2- { the fluoro- 4- of 5- [(pyrido [2,3-d] pyrimidine -6- methylene)-amine]-pyrimidine -2-base ammonia Base }-imidazoles -1- bases)-ethanol
Compound structure：
Synthetic route：
Synthetic method：
Take a certain amount of 2,4-, bis- chloro- 5-FUs to be placed into three-necked flask, add the diisopropylethylamine of 1~5 times of amount And isopropyl alcohol mixed solvent, stir, pyrimido pyridine -6- methylaminos are dissolved in the solution of 1~10 times of amount isopropanol, so After be slowly added in three-necked flask, after adding, -10 DEG C to 30 DEG C react 1 hour~10 hours, be warmed to room temperature naturally, produce A large amount of precipitations are produced, adds methylene chloride and is all dissolved to solid, then with saturated common salt water washing organic matter.Organic matter is with anhydrous After magnesium sulfate is dried, filter off drier and be evaporated, chromatography over CC obtains white product（Chloro- 5-FU -4- the bases of 2-）- pyrido [ 2,3-d] pyrimidine-6- methyl amines, yield 50%~70%.
Take a certain amount of（Chloro- 5-FU -4- the bases of 2-）- pyrido [2,3-d] pyrimidine -6- methyl amines, are placed into three mouthfuls In flask, add the isopropanol of 1~5 times of amount, stir, the air of reaction system is emptied by double Manifold technology, and be filled with nitrogen Gas；1- ethoxys, 2- aminooimidazoles are dissolved in the solution of 1~10 times of amount isopropanol, are added in reaction system, 50 DEG C to 150 DEG C reaction 5 hours~24 hours, after cooling, add ethyl acetate and saturated sodium bicarbonate mixed solution, after stirring 30 minutes, point Go out organic phase, aqueous phase is extracted with ethyl acetate, saturated common salt water washing organic phase and extraction phase.Anhydrous magnesium sulfate is dried.Filter off Drier is evaporated, column chromatography purify white product 2- (2- the fluoro- 4- of 5- [(pyrido [2,3-d] pyrimidine -6- methylene) - Amine]-pyrimidine -2 --amino }-imidazoles-1- bases)-ethanol, yield 50%~80%.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ7.88(1H), δ8.75(1H), δ 7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ8.78(1H), δ9.26(1H)
Embodiment 2：The fluoro- N2- of 5- (1- methyl isophthalic acid H- imidazoles -2- bases)-N4- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases) - Benzyl]-pyrimidine -2,4- diamines
Compound structure：
Synthetic route：
Synthetic method：
Take a certain amount of 2,4-, bis- chloro- 5-FUs to be placed into three-necked flask, add the diisopropylethylamine of 1~5 times of amount And isopropyl alcohol mixed solvent, stir, by 3-（1-methyl isophthalic acid H- pyrazoles-4- bases）Benzylamine is dissolved in 1~10 times and measures isopropanol Solution, then it is slowly added in three-necked flask, after adding, -10 DEG C to 30 DEG C are reacted 1 hour~10 hours, rise to room naturally Temperature, a large amount of precipitations are produced, add methylene chloride and all dissolved to solid, then with saturated common salt water washing organic matter.It is organic After thing is dried with anhydrous magnesium sulfate, filter off drier and be evaporated, chromatography over CC obtains the white product (fluoro- pyrimidine -4- of the chloro- 5- of 2- Base)-[3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-amine, yield 55%~80%.
A certain amount of (the fluoro- pyrimidine-4-yls of the chloro- 5- of 2-)-[3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-amine is taken, is placed Into three-necked flask, add the isopropanol of 1~5 times of amount, stir, the air of reaction system is emptied by double Manifold technology, and It is filled with nitrogen；1- methyl, 2- aminooimidazoles are dissolved in the solution of 1~10 times of amount isopropanol, are added in reaction system, 50 DEG C extremely 150 DEG C are reacted 5 hours~24 hours, after cooling, are added ethyl acetate and saturated sodium bicarbonate mixed solution, are stirred 30 minutes Afterwards, organic phase is separated, aqueous phase is extracted with ethyl acetate, saturated common salt water washing organic phase and extraction phase.Anhydrous magnesium sulfate is dried. Filter off drier to be evaporated, column chromatography purifies to obtain the fluoro- N2- of white product 5- (1- methyl isophthalic acid H- imidazoles -2- bases)-N4- [3- (1- first Base -1H- pyrazoles -4- bases)-benzyl]-pyrimidine -2,4- diamines, yield 50%~70%.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ 7.29(1H), δ7.28(1H), δ7.0(1H), δ6.9(1H), δ3.63(3H), δ8.34(1H), δ8.20(1H), δ 3.80(3H)
Embodiment 3：2- (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2- amino } - Imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H),δ4.0(1-NH),δ4.0(1-NH), δ4.32(2H), δ7.02(2H), δ7.20(1H), δ 7.29(1H), δ7.28(1H), δ8.20(1H), δ8.34(1H), δ3.8(3H), δ7.0(1H), δ7.0(1H), δ 3.92(2H), δ4.01(2H), δ2.0(1-OH)
Embodiment 4：2- (2- { the chloro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2- amino } - Imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.97(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ 7.29(1H), δ7.28(1H), δ8.20(1H), δ8.34(1H), δ3.8(3H), δ7.0(1H), δ7.0(1H), δ 3.92(2H), δ4.01(92H), δ2.0(1-OH)
Embodiment 5：2- (2- { 4- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino] -5- trifluoromethvl-Dvrimidins -2- Base amino }-imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.52(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ 7.29(1H), δ7.28(1H), δ8.40(1H), δ8.40(1H), δ13.7(1NH), δ7.0(1H), δ7.0(1H), δ 3.92(2H), δ4.01(2H), δ2.0(1-OH)
Embodiment 6：2- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino] -4- [3- (1- methyl isophthalic acid H- pyrazoles -4- Base)-benzylamino]-pyrimidine -5- nitriles
Synthetic method is with reference to embodiment 2.
MS：415.19
Embodiment 7：2- (2- { the fluoro- 4- of 5- [the fluoro- 5- of 3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2- Base amino }-imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
MS：426.17
Embodiment 8：2- (2- { 4- [3- (1- ethyl-1H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-miaows Azoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
MS：404.21
Embodiment 9：2- (2- { 4- [(1H- benzimidazole-5- methylene)-amine] the fluoro- pyrimidine -2 --aminos of-5- }-imidazoles- 1- yls)-ethanol
Synthetic method is with reference to embodiment 1.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(1H), δ7.50(1H), δ7.58(1H), δ 7.06(1H), δ7.0(1H), δ7.0(1H) ,δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ8.08(1H), δ 5.0(1NH)
Embodiment 10：2- (2- { 4- [(6,9- dihydro purine-1- methylene)-amine] the fluoro- pyrimidine -2 --aminos of-5- }-miaows Azoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 1.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ3.81(92H), δ7.50(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ7.4(1H), δ13.4(1NH)
Embodiment 11：2- (2- the fluoro- 4- of 5- [(9- hydroxymethyl -6,9- dihydro purine -1- methylene)-amine]-pyrimidine - 2- bases amino }-imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 1.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ3.75(92H), δ7.50(1H), δ7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ7.2(1H), δ5.97(2H), δ 2.0(1-OH)
Embodiment 12：2- [2- (the fluoro- 4- of 5- { 1- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-phenyl]-ethylamino- }-pyrimidine - 2- bases amino)-imidazoles -1- bases]-ethanol
Synthetic method is with reference to embodiment 2.
MS：422.20
Embodiment 13：2- (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-anilino-]-pyrimidine -2-base ammonia Base }-imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ 6.42(1H), δ7.07(1H), δ6.84(1H), δ6.68(1H), δ8.34(1H), δ8.20(1H), δ3.80(3H), δ 4.0(1-NH)
Embodiment 14：2- (2- { the fluoro- 4- of 5- [the fluoro- 5- of 3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-anilino-]-pyrimidine -2-bases Amino }-imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0 (1-OH), δ6.13(1H), δ6.55(1H), δ8.34(1H), δ8.20(1H), δ3.80(3H), δ4.0(1-NH)
Embodiment 15：2- (2- { 4- [3- (1- ethyl -1H- pyrazoles -4- bases)-anilino-] fluoro- pyrimidine -2-base ammonia of -5- Base }-imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
MS：408.18
Embodiment 16：2- { 2- [4- (1H- benzimidazole-5- bases amino) the fluoro- pyrimidine -2 --aminos of-5-]-imidazoles-1- Base }-ethanol
Synthetic method is with reference to embodiment 1.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0 (1-OH), δ6.90(1H), δ7.45(1H), δ6.46(1H), δ4.0(1-NH), δ8.08(1H), δ5.0(1-NH)
Embodiment 17：2- { 2- [the fluoro- 4- of 5- (pyrido [2,3-d] pyrimidine-6- bases amino)-pyrimidine -2 --amino]-miaows Azoles -1- bases }-ethanol
Synthetic method is with reference to embodiment 1.
MS：367.13
Embodiment 18：2- { 2- [4- (6,9- dihydro purine-1- bases amino) the fluoro- pyrimidine -2 --aminos of-5-]-imidazoles-1- Base }-ethanol
Synthetic method is with reference to embodiment 1.
MS：358.14
Embodiment 19：2- { 2- [the fluoro- 4- of 5- (9- hydroxymethyl -6,9- dihydro purine -1- bases amino)-pyrimidine -2-base ammonia Base]-imidazoles -1- bases }-ethanol
Synthetic method is with reference to embodiment 1.
MS：388.15
Embodiment 20：2- [2- (the fluoro- 4- of 5- { methyl-[3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-phenyl]-amine }-pyrimidine - 2- bases amino)-imidazoles -1- bases]-ethanol
Synthetic method is with reference to embodiment 2.
MS：408.18
Embodiment 21：2- { 2- [(the fluoro- 4- of 5- { methyl-[3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-amine }-pyrimidine -2- Base)-methyl-amine]-imidazoles -1- bases }-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ7.29(1H), δ7.28(1H), δ 7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1OH), δ8.34(1H), δ8.20(1H), δ 3.80(3H), δ2.47(3H), δ2.47(3H)
Embodiment 22：2- [2- ({ the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base } - Methyl-amine)-imidazoles -1- bases]-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ7.29(1H), δ 7.28(1H), δ7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ8.34(1H), δ 8.20(1H), δ3.80(3H), δ2.47(3H)
Embodiment 23：2- [2- (the fluoro- 4- of 5- { methyl-[3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-amine }-pyrimidine -2- Base amino)-imidazoles -1- bases]-ethanol
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(91H), δ4.0(1-NH), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ7.29(1H), δ 7.28(1H), δ7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ8.34(1H), δ 8.20(1H), δ3.80(3H), δ2.47(3H)
Embodiment 24：1- { 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine -4- Base amino }-methyl)-phenyl]-pyrazol-1-yl }-ethyl ketone
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4,32(2H), δ7.02(1H), δ7.20(1H), δ 7.29(1H), δ7.28(1H), δ7.0(1H), δ7.0(1H), δ3.92(2H), δ4.01(2H), δ2.0(1-OH), δ 8.4(1H), δ8.4(1H), δ2.20(3H)
Embodiment 25：{ 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine-4-yls Amino }-methyl)-phenyl]-pyrazol-1-yl }-methylsulfonamides
Synthetic method is with reference to embodiment 2.
MS：487.16
Embodiment 26：{ 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine-4-yls Amino }-methyl)-phenyl]-pyrazol-1-yl }-acetic acid
Synthetic method is with reference to embodiment 2.
MS：452.17
Embodiment 27：{ 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine-4-yls Amino }-methyl)-phenyl] pyrazol-1-yl }-acetoxymethyl ester
Synthetic method is with reference to embodiment 2.
MS：466.19
Embodiment 28：2- (2- { the fluoro- 4- of 5- [3- (1- hydroxymethyl -1H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2- Base amino }-imidazoles -1- bases)-ethanol
Synthetic method is with reference to embodiment 2.
MS：424.18
Embodiment 29：(2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-imidazoles -1- bases)-methylsulfonamides
Synthetic method is with reference to embodiment 2.
MS：457.14
Embodiment 30：(2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-imidazoles -1- bases)-acetic acid
Synthetic method is with reference to embodiment 2.
MS：422.16
Embodiment 31：(2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-imidazoles -1- bases)-acetoxymethyl ester
Synthetic method is with reference to embodiment 2.
MS：436.18
Embodiment 32：(2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-imidazoles -1- bases)-methanol
Synthetic method is with reference to embodiment 2.
MS：394.17
Embodiment 33：The fluoro- N4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-N2- (4- methyl -4H- [1,2,4] three Azoles -3- bases)-pyrimidine -2,4- diamines
Synthetic method is with reference to embodiment 2.
MS：379.17
Embodiment 34：(3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazole-4-yl)-methylsulfonamides
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ 7.29(1H), δ7.28(1H), δ7.0(1H), δ6.9(1H), δ8.34(1H), δ8.20(1H), δ3.80(3H), δ 5.85(2H), δ2.0(2-NH)
Embodiment 35：(3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazole-4-yl)-acetic acid
Synthetic method is with reference to embodiment 2.
1H-NMR(400MHZ,CDCl3, TMS, ppm):
δ7.44(1H), δ4.0(1-NH), δ4.0(1-NH), δ4.32(2H), δ7.02(1H), δ7.20(1H), δ 7.29(1H), δ7.28(1H), δ7.0(1H), δ6.9(1H), δ8.34(1H), δ8.20(1H), δ3.80(3H), δ 4.67(2H), δ11.0(1-OH)
Embodiment 36：(3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazole-4-yl)-acetoxymethyl ester
Synthetic method is with reference to embodiment 2.
MS：437.17
Embodiment 37：(3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazole-4-yl)-methanol
Synthetic method is with reference to embodiment 2.
MS：395.16
Embodiment 38：The fluoro- N4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-N2- (2- methyl -2H- [1,2,4] three Azoles -3- bases)-pyrimidine -2,4- diamines
Synthetic method is with reference to embodiment 2.
MS：379.17
Embodiment 39：(5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazol-1-yl)-methylsulfonamides
Synthetic method is with reference to embodiment 2.
MS：458.14
Embodiment 40：(5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazol-1-yl)-acetic acid
Synthetic method is with reference to embodiment 2.
MS：423.16
Embodiment 41：(5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazol-1-yl)-acetoxymethyl ester
Synthetic method is with reference to embodiment 2.
MS：437.17
Embodiment 42：(5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base ammonia Base }-[1,2,4] triazol-1-yl)-methanol
Synthetic method is with reference to embodiment 2.
MS：395.16
Embodiment 43：With the binding tests of JAK3 targets
Compound listed by the embodiment of the present invention 1~42, Tofacitinib and Decernotinib are entered with JAK3 respectively Row molecular docking, observe the combination situation of itself and JAK3 targets.It the results are shown in Table 1.
The Binding experiment result of table 1 and JAK3 targets
Note：1.（A）80 ＜ and JAK3 targets conjugation ＜ 100；
2.（B）100 ＜ and JAK3 targets conjugation ＜ 120；
3.（C）120 ＜ and JAK3 targets conjugation ＜ 140；
4.（D）With the conjugation ＞ 140 of JAK3 targets.
Embodiment 44：With the binding tests of STAT3 targets
By the compound listed by the embodiment of the present invention 1~42, Tofacitinib, Filgotinib and Decernotinib Molecular docking is carried out with STAT3 respectively, observes the combination situation of itself and STAT3 targets.It the results are shown in Table 2.
The Binding experiment result of table 2 and STAT3 targets
Note：1.（A）80 ＜ and STAT3 targets conjugation ＜ 100；
2.（B）100 ＜ and STAT3 targets conjugation ＜ 120；
3.（C）120 ＜ and STAT3 targets conjugation ＜ 140；
4.（D）With the conjugation ＞ 140 of STAT3 targets.
Embodiment 45：To JAK inhibitory action
Influence of the compound to the restructuring JAK activity of purifying is studied, is the suppression from the horizontal research compound of zymetology to JAK Activity.Its experimental principle is to use a kind of luminescence method kinase assay, for detecting JAK and substrate Poly (4:1 Glu, Tyr) ADP contents caused by reactive polypeptide：After ADP is converted into ATP, ATP can be used as Ultra-Glo luciferase catalytic reactions Substrate, produce optical signal.Luminous signal and ADP amount and kinase activity positive correlation.Therefore, by observing compound to JAK With substrate reactions caused by luminous signal determine its JAK to restructuring inhibition, use IC50Represent.
Experimental method：The compound of 10 various concentrations respectively 37 DEG C with JAK1, JAK2 and JAK3 be incubated 60 minutes, so Afterwards add substrate and ATP mixing, 37 DEG C reaction 50 minutes after add 25 μ lADP-Glo mix 2 minutes, react at room temperature 50 points Clock.Add 50 μ l detection reagents to mix 2 minutes, be incubated at room temperature 50 minutes, detected with Chemiluminescence Apparatus.It the results are shown in Table 3.
Inhibitory action experimental result of the table 3 to JAK
Note：1.（A）20nM or smaller；
2.（B）>20nM to 100nM；
3.（C）> 100nM
Embodiment 46：To p-STAT5 inhibitory action
JAK-STAT signal transmission paths are occurred mainly in leucocyte, therefore participate in immunological regulation.IL-3 activation is thin Cause JAK2 that autophosphorylation occurs after acceptor on after birth and activate.Stat protein is incorporated on the acceptor of phosphorylation and quilt JAK phosphorylations.The STAT of phosphorylation and the stat protein of another phosphorylation combine to form dimer and to cell nuclear transfers. In nucleus, STAT is combined with DNA and is promoted genetic transcription, causes immune response.Therefore, IL-3 is situated between by observing compound The STAT5 led（It is as shown in the table）Phosphorylation determine its inhibition to JAK2.
Experimental method：Compound is diluted according to 11 various concentrations with DMSO, takes the compound after 200 μ l dilutions to add 24 orifice plates；By TF-1 cells（ATCC TIB-202）With 0.5% serum starvation overnight, 2 × 10 are then adjusted to6/ ml, Take 200 μ l cells be added to it is above-mentioned have been added in 24 orifice plates of compound, after mixing, 37 DEG C of cell culture incubators are incubated 60 points Clock；After 60 minutes, 12ng IL-3 is added per hole, 37 DEG C of cell culture incubators stimulate 60 minutes again；After stimulating 60 minutes, 2%PFA Room temperature fixes 30 minutes；Cell after fixation is added in BD streaming pipes, 4 DEG C of horizontal centrifugals, 1500rpm, 5 minutes；Add 500 μ l methanol, 4 DEG C of incubations wear film in 60 minutes；4 DEG C of horizontal centrifugals, 1500rpm, 5 minutes；Add 2mlFACS buffer, 4 DEG C Horizontal centrifugal, 1500rpm, 5 minutes；Addition 2mlFACS buffer, 4 DEG C of horizontal centrifugals, 1500rpm, 5 minutes；Often pipe adds Enter 5 μ l STAT5（pY694）Antibody（562077）, room temperature lucifuge incubation 60 minutes；Add 2mlFACS buffer, 4 DEG C of levels Centrifugation, 1500rpm, 5 minutes；Often pipe adds 300 μ lFACS buffer and is resuspended, and with flow cytomery, observes compound pair P-STAT5 inhibitory action, uses IC50Represent.It the results are shown in Table 4.
Inhibitory action experimental result of the table 4 to p-STAT5
Note：1.（A）200nM or smaller；
2.（B）>200nM to 2000nM；
Embodiment 47：To p-STAT6 inhibitory action
IL-4 inductions STAT6 phosphorylation is the key for detecting inhibitor in the cellular level activity of JAK1-JAK3 paths Experiment.
Experimental method：Compound is diluted according to 11 various concentrations with DMSO, takes the compound after 200 μ l dilutions to add 24 orifice plates；By THP1 cells（ATCC TIB-202）It is adjusted to 2 × 106/ ml, 200 μ l cells are taken to be added to above-mentioned have been added to In 24 orifice plates of compound, after mixing, 37 DEG C of cell culture incubators are incubated 60 minutes；After 60 minutes, 8ng IL-4 is added per hole, 37 DEG C of cell culture incubators stimulate 60 minutes again；After stimulating 60 minutes, 30 minutes are fixed with 2%PFA room temperatures；By the cell after fixation It is added in BD streaming pipes, 4 DEG C of horizontal centrifugals, 1500rpm, 5 minutes；500 μ l methanol are added, 4 DEG C of incubations wear film in 60 minutes；4 DEG C horizontal centrifugal, 1500rpm, 5 minutes；Addition 2mlFACS buffer, 4 DEG C of horizontal centrifugals, 1500rpm, 5 minutes；Add 2mlFACS buffer, 4 DEG C of horizontal centrifugals, 1500rpm, 5 minutes；Often pipe adds 5 μ lSTAT6（pY641）Antibody （562078）, room temperature lucifuge incubation 60 minutes；Addition 2mlFACS buffer, 4 DEG C of horizontal centrifugals, 1500rpm, 5 minutes；Often Pipe adds 300 μ lFACS buffer and is resuspended, and with flow cytomery, observation compound is used p-STAT6 inhibitory action IC50Represent.It the results are shown in Table 5.
Inhibitory action experimental result of the table 5 to p-STAT6
Note：1.（A）200nM or smaller；
2.（B）>200nM to 2000nM；
Embodiment 48：The influence of the mouse rheumatoid arthritis induced collagen
In the mouse rheumatoid arthritis that collagen induces（CIA）In model, the suppression for studying the compound of selection is lived Property.TypeⅡ Collagen（CII）It is present in mostly in articular cartilage, is a kind of protein with immune system isolation, but in disease It can be showed under the conditions of reason as a kind of autoantibody, 50% rheumatoid arthritis（RA）Have anti-CII's in patients serum Autoantibody, this shows that CII can induce the autoimmune response for producing arthritis property.
Secondary immunity, initial immunity are carried out with 10 week old male DBA-1 mouse（On 0th）：Cotton ball soaked in alcohol sterilizes mouse Root of the tail portion skin, the μ l of collagen 100 emulsified are subcutaneously injected at the 2-3cm of mouse tail root（CII and CFA volume ratios are 1:1）（Containing the μ g of CII 150 and the μ g of heat-inactivated mycobacteria 50）；Secondary immunity（On 21st）：Cotton ball soaked in alcohol sterilizes mouse tail Root skin, the μ l of collagen 50 emulsified are subcutaneously injected at the 2-3cm of mouse tail root（CII is 1 with IFA volume ratios:1） （Containing the μ g of CII 75）.Mouse is grouped at random after secondary immunity, is divided into control group (cosolvent group) and test group.Test group is by chemical combination Thing is suspended in the water slurry of 1% methylcellulose, starts to be administered to the administration of the 41st end of day on 23rd, drug dose is 30mg/kg, each μ l of gavage 200,2 times a day；Control group gives cosolvent with method.Daily test and appraisal clinical inflammatory symptom score. Standards of grading are as follows：0 point：Without erythema and oedema；1 point：Two sufficient little toe arthritic erythema oedema；2 points：Whole toe joints or front foot Palman erythema oedema；3 points：Ankle-joint extends below toe joint erythema oedema；4 points：Ankle-joint is to full pawl is red and swollen or joint deformity. Every mouse four limbs scoring is added general comment and is divided into mouse arthritis index, and total score is 16 points.Control group and test group are observed respectively Mouse inflammatory symptom, clinical inflammatory symptom score is calculated, using bilateral, unpaired t-test method comparative test group and control group Clinical inflammatory symptom score, the influence for the mouse rheumatoid arthritis that evaluation compound is induced collagen, is represented with P values. It the results are shown in Table 6.
The influence for the mouse rheumatoid arthritis that table 6 is induced collagen
Note：1.P ＜ 0.05, test group have aobvious with control group mice clinical inflammatory symptom score statistical analysis comparing difference Work property；
2.P ＜ 0.01, test group have extremely aobvious with control group mice clinical inflammatory symptom score statistical analysis comparing difference Work property.
Embodiment 49：To the inhibitory action of human tumor cells in-vitro multiplication
Take gastric carcinoma cells SNU-5, liver cancer cells Hep-G2, lung carcinoma cell EBC-1, stomach cancer cell BGC-823, brain star Glioblastoma U87MG, cervical cancer cell Hela, breast carcinoma Bcap-37, compound is configured to 20mM's with DMSO Solution, by series compound and taxol（Liquid storage 0.2mM）With 3 times of gradient dilutions of DMSO（10 concentration）；Take 5 μ l terraced respectively Spend the compound solution diluted and taxol is added to 495 μ l and contained in 10% FBS culture medium, be configured to 2X test compounds Thing.
100 μ l testing compounds containing 2X, taxol are taken to be added in 96 orifice plate respective apertures, carbon dioxide cell incubator culture 72 hours.
Culture medium is removed, XTT working solutions are added per hole（0.3mg/ml XTT；0.00265mg/ml PMS）150 μ l, dioxy Change and placed 2 hours in carbon incubator, micropore plate oscillator is shaken 5 minutes, and ELIASA 450nm reads light absorption value, calculates compound To the inhibiting rate of human tumor cells（%）, try to achieve IC50Value（μM）.It the results are shown in Table 7.
Inhibitory action of the table 7 to human tumor cells in-vitro multiplication（IC50, μM）
Embodiment 50：Pharmacokinetics and bioavailability study
Compound is taken, healthy mice single oral gastric infusion, dosage 30mg/kg, takes mice serum, determines Related Drug For kinetic parameter, and compared with Tofacitinib, relative bioavailability F is calculated.It the results are shown in Table 8.
Compound is taken, the administration of healthy mice single intravenous injection, dosage is 5 mg/kg, takes mice serum, determines Related Drug For kinetic parameter, and compared with Tofacitinib, relative bioavailability F is calculated.It the results are shown in Table 9.
Tested in embodiment 48 and mice serum is taken within 1 hour after closing day last time is administered, measure Related Drug is for power Parameter is learned, and compared with Tofacitinib, calculates relative bioavailability F.It the results are shown in Table 10.
The healthy mice single oral gastric infusion relative bioavailability measurement result of table 8
Note：1.（A）Relative bioavailability is 90%~100% compared with Tofacitinib；
2.（B）Relative bioavailability is 100%~120% compared with Tofacitinib.
Relative bioavailability measurement result is administered in the healthy mice single intravenous injection of table 9
Note：1.（A）Relative bioavailability is 90%~100% compared with Tofacitinib；
2.（B）Relative bioavailability is 100%~120% compared with Tofacitinib.
The rheumatoid arthritis mouse relative bioavailability measurement result that the collagen of table 10 induces
Note：1.（A）Relative bioavailability is 90%~100% compared with Tofacitinib；
2.（B）Relative bioavailability is 100%~120% compared with Tofacitinib.
Embodiment 51：Cell toxicity test
Experimental principle：Wst-8 in CCK8, can be by Intramitochondrial dehydrogenation in the presence of electronics coupled reagent The orange-yellow formazan product of enzyme reduction generation high water soluble（formazan）.The depth of color and being proliferated into just for cell Than being inversely proportional with cytotoxicity.OD values are determined at 450nm wavelength using ELIASA, reflects the quantity of living cells indirectly, is used for Determine the cytotoxicity of compound.
Experimental method：According to flow cytometer in experimentation and microscopical morphologic observation, each toxicity of compound is very It is small.In order to further quantitatively confirm toxicity of compound, the propagation of HELA cells is have detected with CCK-8 method, method detailed is such as Under：Compound is diluted according to 10 various concentrations with DMSO, takes the compound after 100ml dilutions to add 96 orifice plates；By HELA Cell is adjusted to 2 × 105/ ml, take 100ml cells be added to it is above-mentioned have been added in 96 orifice plates of compound, after mixing, 37 DEG C Cell culture incubator is incubated 24 hours；20 μ l CCK solution are added to every hole；Culture plate is incubated 4 hours in incubator；Use enzyme Mark absorbance of the instrument measure at 450nm.
Experimental result：Each compound changes cell quantity with not having concentration dependent, shows these compounds to HELA Cytotoxic.It the results are shown in Table 11.
The cytotoxicity experiment result of table 11
Embodiment 52：Rat acute toxicity test
Compound is taken, observes rat single oral gastric infusion acute toxicity, calculates LD50.It the results are shown in Table 12.
The rat single oral gastric infusion acute toxicity testing result of table 12
Note：1.（A）LD50＜ 10mg/kg；
2.（B）10mg/kg ＜ LD50＜ 50mg/kg；
3.（C）50mg/kg ＜ LD50＜ 100mg/kg；
4.（D）100mg/kg ＜ LD50＜ 500mg/kg；
5.（E）500mg/kg ＜ LD50＜ 1000mg/kg；
6.（F）LD50＞ 1000mg/kg.
Described above is only the preferred embodiment of the present invention, for those skilled in the art, Without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should be regarded as this hair Bright protection domain.

Claims (4)

  1. A kind of 1. compound of formula (I)：
    Or its pharmaceutically acceptable salt, wherein：
    Ring A is C3-7Cycloalkyl, C5-7Fragrant ring group, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, Its middle ring A is optionally by one or more identical or different R, R1Substituted；
    R、R1It is H, halogen, C3-7Cycloalkyl, C5-7Fragrant ring group, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11Fragrance is miscellaneous Bicyclic group, optionally by one or more identical or different R wherein on these rings5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynes Base, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl is optionally by one or more identical or different R6Substitution；
    B1It is H, CH3、CN、NO2、CF3, halogen；
    B2It is H, CH3、CN、NO2、CF3, halogen；
    X is H, CH3、CN、NO2、CF3、C(O)NH2, halogen；
    R2It is H, CH3、CN、NO2、CF3, halogen；
    R3It is H, CH3、CN、NO2、CF3, halogen；
    R4It is H, CN, NO2、CF3、COOH、COOR7, halogen；C3-7Cycloalkyl, C5-7Fragrant ring group, C5-7Fragrant heterocyclic radical, C7-11Virtue Fragrant bicyclic group, C7-11The miscellaneous bicyclic group of fragrance, optionally by one or more identical or different R wherein on these rings5Substitution；C1-8 Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl is optionally by one or more identical or different R6Substitution；
    R5It is H, CN, NO2、CF3、COOH、COOR7, halogen；
    R6It is H, CN, NO2、CF3、COOH、COOR7, halogen；
    R7It is C3-7Cycloalkyl, C5-7Fragrant ring group, C5-7Fragrant heterocyclic radical, C7-11Aromatic bicyclic base, C7-11The miscellaneous bicyclic group of fragrance, its In on these rings optionally by one or more identical or different R5Substitution；C1-8Alkyl, C2-8Alkenyl, C2-8Alkynyl, wherein C1-8 Alkyl, C2-8Alkenyl, C2-8Alkynyl is optionally by one or more identical or different R6Substitution；
    Y is (CR12R13)n；
    N is 0 or 1；
    R12、R13It is R5；
    Z1、Z2C (R can be respectively selected from14) or N (R14)；
    R14It is H, CN, NO2、CF3、COOH、COOR7, halogen.
  2. 2. following compound：
    2- (2- { the fluoro- 4- of 5- [(pyrido [2,3-d] pyrimidine-6- methylene)-amine]-pyrimidine -2 --amino }-imidazoles-1- bases)- Ethanol；
    The fluoro- N2- of 5- (1- methyl isophthalic acid H- imidazoles -2- bases)-N4- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-pyrimidine -2,4- two Amine；
    2- (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2- amino }-imidazoles -1- bases) - Ethanol；
    2- (2- { the chloro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2- amino }-imidazoles -1- bases) - Ethanol；
    2- (2- { 4- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino] -5- trifluoromethvl-Dvrimidin -2- bases amino }-miaows Azoles -1- bases)-ethanol；
    2- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino] -4- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl ammonia Base]-pyrimidine -5- nitriles；
    2- (2- { the fluoro- 4- of 5- [the fluoro- 5- of 3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-imidazoles- 1- yls)-ethanol；
    2- (2- { 4- [3- (1- ethyl-1H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-imidazoles-1- bases)-second Alcohol；
    2- (2- { 4- [(1H- benzimidazole-5- methylene)-amine] the fluoro- pyrimidine -2 --aminos of-5- }-imidazoles-1- bases)-ethanol；
    2- (2- { 4- [(6,9- dihydro purine-1- methylene)-amine] the fluoro- pyrimidine -2 --aminos of-5- }-imidazoles-1- bases)-second Alcohol；
    2- (2- { the fluoro- 4- of 5- [(9- hydroxymethyl-6,9- dihydro purine-1- methylene)-amine]-pyrimidine -2 --amino }-miaows Azoles -1- bases)-ethanol；
    2- [2- (the fluoro- 4- of 5- { 1- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-phenyl]-ethylamino- }-pyrimidine -2 --amino)-miaows Azoles -1- bases]-ethanol；
    2- (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-anilino-]-pyrimidine -2 --amino }-imidazoles-1- bases)- Ethanol；
    2- (2- { the fluoro- 4- of 5- [the fluoro- 5- of 3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-anilino-]-pyrimidine -2 --amino }-imidazoles-1- Base)-ethanol；
    2- (2- { 4- [3- (1- ethyl-1H- pyrazoles-4- bases)-anilino-] the fluoro- pyrimidine -2 --aminos of-5- }-imidazoles-1- bases)- Ethanol；
    2- { 2- [4- (1H- benzimidazole-5- bases amino) the fluoro- pyrimidine -2 --aminos of-5-]-imidazoles-1- bases }-ethanol；
    2- { 2- [the fluoro- 4- of 5- (pyrido [2,3-d] pyrimidine-6- bases amino)-pyrimidine -2 --amino]-imidazoles-1- bases }-ethanol；
    2- { 2- [4- (6,9- dihydro purine-1- bases amino) the fluoro- pyrimidine -2 --aminos of-5-]-imidazoles-1- bases }-ethanol；
    2- { 2- [the fluoro- 4- of 5- (9- hydroxymethyl-6,9- dihydro purine-1- bases amino)-pyrimidine -2 --amino]-imidazoles-1- Base }-ethanol；
    2- [2- (the fluoro- 4- of 5- { methyl-[3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-phenyl]-amine }-pyrimidine -2 --amino)-miaows Azoles -1- bases]-ethanol；
    2- 2- [(the fluoro- 4- of 5- { methyl-[3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-amine }-pyrimidine -2-base)-methyl-amine] - Imidazoles -1- bases }-ethanol；
    2- [2- ({ the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzylamino]-pyrimidine -2-base }-methyl-amine)-miaows Azoles -1- bases]-ethanol；
    2- [2- (the fluoro- 4- of 5- { methyl-[3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzyl]-amine }-pyrimidine -2 --amino)-imidazoles- 1- yls]-ethanol；
    1- 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine-4-yl amino }-methyl) - Phenyl]-pyrazol-1-yl }-ethyl ketone；
    { 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine-4-yl amino }-methyl)-benzene Base]-pyrazol-1-yl }-methylsulfonamides；
    { 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine-4-yl amino }-methyl)-benzene Base]-pyrazol-1-yl }-acetic acid；
    { 4- [3- ({ the fluoro- 2- of 5- [1- (2- hydroxy-ethyls) -1H- imidazoles -2- bases amino]-pyrimidine-4-yl amino }-methyl)-benzene Base] pyrazol-1-yl }-acetoxymethyl ester；
    2- (2- { the fluoro- 4- of 5- [3- (1- hydroxymethyl-1H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-imidazoles- 1- yls)-ethanol；
    (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-imidazoles-1- bases)- Methylsulfonamides；
    (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-imidazoles-1- bases)- Acetic acid；
    (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-imidazoles-1- bases)- Acetoxymethyl ester；
    (2- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-imidazoles-1- bases)- Methanol；
    The fluoro- N4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-N2- (4- methyl -4H- [1,2,4] triazole -3- bases)-phonetic Pyridine -2,4- diamines；
    (3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 4- yls)-methylsulfonamides；
    (3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 4- yls)-acetic acid；
    (3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 4- yls)-acetoxymethyl ester；
    (3- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 4- yls)-methanol；
    The fluoro- N4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles -4- bases)-benzyl]-N2- (2- methyl -2H- [1,2,4] triazole -3- bases)-phonetic Pyridine -2,4- diamines；
    (5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 1- yls)-methylsulfonamides；
    (5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 1- yls)-acetic acid；
    (5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 1- yls)-acetoxymethyl ester；
    (5- { the fluoro- 4- of 5- [3- (1- methyl isophthalic acid H- pyrazoles-4- bases)-benzylamino]-pyrimidine -2 --amino }-[1,2,4] triazole- 1- yls)-methanol.
  3. 3. compound as claimed in claim 1 or 2 is prevented in human patient is prepared or treated and Janus kinases (JAK) phase Applied in the disease of pass and the medicine of illness.
  4. 4. compound as claimed in claim 1 or 2 is prevented in mammalian subject is prepared or treated and Janus kinases (JAK) applied in the medicine of related disease and illness.
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