CN109422753A

CN109422753A - One kind has inhibition and the active compound of tyrosine protein kinase JAK1 or JAK2 of degrading

Info

Publication number: CN109422753A
Application number: CN201710783082.5A
Authority: CN
Inventors: 舒永志
Original assignee: Shanghai Zhi Zhi Medical Science And Technology Co Ltd
Current assignee: Shanghai Zhi Zhi Medical Science And Technology Co Ltd
Priority date: 2017-09-03
Filing date: 2017-09-03
Publication date: 2019-03-05
Anticipated expiration: 2037-09-03
Also published as: CN109937203B; CN109937203A; WO2019042442A1; CN109422753B

Abstract

The present invention provides one kind to have inhibition and the active compound of tyrosine protein kinase JAK1 or JAK2 of degrading, and specifically, the present invention provides a kind of such as following formula I compound represented；Wherein, the definition of each group is as noted in the discussion.The compound of the present invention has good JAK1 or JAK2 inhibition and degrading activity, can be used for preparing the drug of the relevant disease of JAK1 or JAK2 between treatment.

Description

One kind has inhibition and tyrosine protein kinase JAK1 or JAK2 activeization of degrading Close object

Technical field

The invention belongs to field of medicaments, have more particularly to one kind inhibit and degrade tyrosine protein kinase JAK1 or The active compound of JAK2 and its preparation and application.

Background technique

Janus kinases (JAKs) is to be present in intracellular non-receptor tyrosine protein kinase, to T cell growth, activation It is particularly significant with the multiple functions such as steady-state adjustment, it plays a crucial role in adjusting lymphohematological cell function.Mammal In, which contains 4 principal home's family members, i.e. JAK1, JAK2, JAK3 and Tyk2.Wherein, JAK1, JAK2 and Tyk2 The wide expression in various histocytes, the main high efficient expression in hematopoietic tissue of JAK3, the bone-marrow-derived lymphocyte, T lymph of such as activation Cell, bone marrow cell and thymocyte etc..

JAK1 can be with IL-10, IL-19, IL-20, IL-22, IL-26, IL-28, IFN-α, IFN-γ, in gp130 family IL-6 and other receptors etc. of the c containing γ combine.JAK1 gene knockout experiment on mouse model shows the enzyme in adjusting It states in the biological effect of cytokine profiles receptor and plays key effect.JAK1 has become the diseases such as immune, inflammation and cancer The novel target spot in field.

JAK2 is including EPO, GH, PRL, the IL-3 in IFN-γ and β c family, a variety of receptors letters such as IL-5, GM-CSF It plays a significant role in number adjustment process.JAK2 is knocked out in mouse model can lead to animal dead caused by anaemia.In human body JAK2 gene on a base mutation JAK2V617F, with the polycythemia vera in myeloproliferative disease (PV), the hair of essential thrombocythemia (ET), idiopathic myelofibrosis (IMF), chronic myelocytic leukemia (CML) etc. Life is closely related.Therefore, JAK2 has become the definite action target spot of such disease treatment and prevention.

It is organ-graft refection, heterograft, lupus erythematosus, more due to the extensive adjustment effect of JAK-STAT signal path Hair property hardening, rheumatoid arthritis, psoriasis (psoriasis), cancer, asthma, atopic dermatitis, type-1 diabetes mellitus and diabetes Complication, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer disease, leukaemia, lymph Many diseases such as tumor, Huppert's disease, alopecia areata, leucoderma also relate to JAK1/2, and JAK1/2 is transmitted as signal Member plays a crucial role in disease generation, becomes the drug targets that this kind of disease is treated in exploitation in medicine research and development.

Therefore, those skilled in the art, which are dedicated to developing, is able to suppress the active compound of JAK1 and/or JAK2.

Summary of the invention

It is an object of the invention to provide a kind of compounds being able to suppress and degrade JAK1 and/or JAK2, and its preparation And application.

In the first aspect of the present invention, provides one kind such as following formula I compound represented or its is pharmaceutically acceptable Salt:

Wherein:

Indicate singly-bound；

Indicate singly-bound or double bond；

A missing is selected from C (=O), C (=O) X1, SOX1, SO₂The C of X1, with or without substituent group_1-8Alkyl, band Have or without substituent group C_1-8The C of cyclic hydrocarbon radical and with or without substituent group_1-8Heterocyclic hydrocarbyl；Wherein X1 missing or choosing From (CR₁₂R₁₃)_fO、(CR₁₂R₁₃)_fS and NR₁₄；Wherein R₁₂、R₁₃、R₁₄It is independent for H, with or without substituent group C_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical and with or without substituent group C_1-8Heterocyclic hydrocarbon Base, f are the integer (such as 1,2 or 3) between 0 to 3；

W missing is selected from O, NR₁₇,-X2C (=O) X3 ,-X2S (=O)_gX3；Wherein R₁₇For H, with or without substituted The C of base_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbon Base；Wherein X2, X3 are independent lacks or is selected from O, S, NR₁₈；The wherein integer that g is 0 to 2；Wherein R₁₈For H, have or not C with substituent group_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8 Heterocyclic hydrocarbyl；

Y is (CR₂₂R₂₃)_h、CHX4(CR₂₂R₂₃)_h, CX4=CH (CR₂₂R₂₃)_hOr (CR₂₂R₂₃)_h；Wherein h is between 0 to 30 Integer；Wherein R₂₂、R₂₃It is independently selected from the C of H, cyano, hydroxyl, amino, with or without substituent group_1-8Hydrocarbon The C of base, with or without substituent group_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl, have or not C with substituent group_1-8Oxyl；Wherein X4 is selected from H, halogen, cyano, nitro, hydroxyl, with or without substituent group C_1-8 Oxyl, with or without substituent group C_1-8Hydrocarbon carbonyl oxygen, with or without substituent group C_1-8Amido, with or without take For base C_1-8Ester group, with or without substituent group C_1-8Amino-carbonyl, with or without substituent group C_1-8Alkyl has or not With substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl；

Z is (CR₂₄R₂₅)_i、CHX5(CR₂₄R₂₅)_i, CX5=CH (CR₂₄R₂₅)_iOr C ≡ C (CR₂₄R₂₅)_i；Wherein i is 0 to 30 Between integer；Wherein R₂₄、R₂₅It is independently selected from the C of H, cyano, hydroxyl, amino, with or without substituent group_1-8's Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl, with or without take For base C_1-8Oxyl；Wherein X5 is selected from H, halogen, cyano, nitro, hydroxyl, with or without substituent group C_1-8Oxyl, With or without substituent group C_1-8Hydrocarbon carbonyl oxygen, with or without substituent group C_1-8Amido, with or without substituent group C_1-8 Ester group, with or without substituent group C_1-8Amino-carbonyl, with or without substituent group C_1-8Alkyl, with or without substituted Base C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl；

B missing is selected from O, C=O, S, NR₁₅、-NR₁₅C (=O)-,-C (=O) NR₁₅,-C (=O) O-, OC (=O) O-、-NR₁₅C (=O) O- ,-OC (=O) NR₁₅-、-NR₁₅C (=O) NR₁₆, with or without substituent group C_1-12Alkyl, The C of with or without substituent group_1-12Cyclic hydrocarbon radical and with or without substituent group C_1-12Heterocyclic hydrocarbyl；Wherein R₁₅、 R₁₆It is independently selected from the C of H, with or without substituent group_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon The C of base and with or without substituent group_1-8Heterocyclic hydrocarbyl；

X is selected from CR₁₉R₂₀, C (=O), S (=O), SO₂、NR₂₁；Wherein R₁₉、R₂₀Be independently selected from H, cyano, hydroxyl, The C of amino, with or without substituent group_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without The C of substituted base_1-8Heterocyclic hydrocarbyl, with or without substituent group C_1-8Oxyl；Wherein R₂₁Selected from H, have or not C with substituent group_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8 Heterocyclic hydrocarbyl；

R₁、R₈It is each independently selected from the C of H, with or without substituent group_1-8Alkyl, with or without substituent group Cyclic hydrocarbon radical, the heterocyclic hydrocarbyl of with or without substituent group, with or without substituent group C_1-6Acyl group；

R₂、R₅It is independently selected from: hydrogen, OR₃₃、NR₃₄R₃₅, cyano, halogen, with or without substituent group C_1~8Hydrocarbon Base, the cyclic hydrocarbon radical of with or without substituent group, the heterocyclic hydrocarbyl of with or without substituent group, with or without substituent group C_1-6The amide groups of acyl group, with or without substituent group；Wherein R₃₃、R₃₄、R₃₅Be independently selected from H, with or without The C of substituted base_1~8Alkyl, with or without substituent group cyclic hydrocarbon radical, the heterocyclic hydrocarbyl of with or without substituent group；

R₃、R₆、R₇、R₉、R₁₀It is independently selected from: H, OR₂₇、NR₂₈R₂₉, cyano, halogen, nitro, with or without The C of substituent group_1-8Alkyl, the cyclic hydrocarbon radical of with or without substituent group, the heterocyclic hydrocarbyl of with or without substituent group, X6S (=O)_kR₃₀, X6C (=O) R₃₁；Wherein k is the integer between 0 to 2；Wherein R₂₇, R₂₈, R₂₉, R₃₀, R₃₁It is independently selected from H, the C of with or without substituent group_1~8Alkyl, cyclic hydrocarbon radical, heterocyclic hydrocarbyl；Wherein X6 lacks or is selected from O, S, NR₃₂；Wherein R₃₂ For H, the C of with or without substituent group_1-8Alkyl, with or without substituent group cyclic hydrocarbon radical and with or without take The heterocyclic hydrocarbyl of Dai Ji；

R₄Selected from H, cyano, carboxyl, with or without substituent group C_1-8Alkyl, with or without substituent group Hydrocarbon carbonyl oxygen；

A is the integer (such as 1,2,3,4,5) between 0 to 5；

B is the integer (such as 1,2,3) between 0 to 3；

C is the integer (such as 1,2,3,4,5,6,7,8,9) between 0 to 30；

D is the integer (such as 1,2,3,4,5) between 0 to 5；

E is the integer (such as 0,1,2) between 0 to 2.

In another preferred example, A is lacked；W is X2C (=O) X3, wherein X2 NR₁₈And X3 missing or X3 are NR₁₈And X2 is lacked；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from H, hydroxyl, with or without substituent group C_1-4Alkyl, h be 1 to 6 between integer；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from H, hydroxyl, has Or the C without substituent group_1-4Alkyl, i be 1 to 6 between integer；C is 0.

In another preferred example, A is lacked；W missing is O；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from H, the C of hydroxyl, with or without substituent group_1-4Alkyl, h be 0 to 3 between integer；B is O；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4Alkyl, i be 0 to 3 between integer；c For the integer between 1 to 6.

In another preferred example, A is C (=O) X1, and wherein X1 lacks or be selected from (CR₁₂R₁₃)_fAnd (CR O,₁₂R₁₃)_fS, Middle f is the integer between 0 to 2, R₁₂、R₁₃、R₁₄Independent is hydrogen or C_1-4Alkyl；W is NR₁₇, wherein R₁₇For H or band Have or without substituent group C_1-4Alkyl；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃Be independently selected from H, hydroxyl, have or Without the C of substituent group_1-4Alkyl, h be 0 to 3 between integer；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅Choosing independent From H, hydroxyl, with or without substituent group C_1-4Alkyl, i be 0 to 3 between integer；B is O；C is whole between 1 to 4 Number.

In another preferred example, A SO₂X1, wherein X1 lacks or is selected from O and S；W is O；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4Alkyl, h be 1 to 6 between integer；Z For (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4Alkyl, i 0 Integer between to 3；C is 0.

In another preferred example, A is lacked；W is NR₁₇, wherein R₁₇For the H or C of with or without substituent group_1-4Hydrocarbon Base；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4Alkyl, h For the integer between 0 to 3；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from H, hydroxyl, with or without substituted The C of base_1-4Alkyl, i be 0 to 4 between integer；B is O；C is the integer between 1 to 6.

In another preferred example, A is lacked；W missing；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from H, hydroxyl The C of base, with or without substituent group_1-4Alkyl, h be 0 to 3 between integer；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅Respectively The independent C for being selected from H, hydroxyl, with or without substituent group_1-4Alkyl, i be 0 to 3 between integer；B is O；C be 1 to Integer between 10 (between preferably 1 to 6).

In another preferred example, any substituent group is selected from the group: halogen, unsubstituted or halogenated C1-C6 alkyl, Unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or halogenated C2-C6 alkoxyalkyl, unsubstituted or halogenated C3-C8 ring Alkyl, unsubstituted or halogenated C2-C6 alkyl-carbonyl, unsubstituted or halogenated C1-C6 alkylidene-hydroxyl, unsubstituted or C1-C6 Alkyl-substituted amido.

In another preferred example, in Formulas I,Indicate singly-bound.

In another preferred example, the X is C (=O).

In another preferred example, R₁It is selected from: the C of H and with or without substituent group_1-4Alkyl.

In another preferred example, R₂、R₅It is independently selected from: the C of hydrogen and with or without substituent group_1-4Alkyl.

In another preferred example, R₃It is selected from: the C of hydrogen and with or without substituent group_1-4Alkyl.

In another preferred example, R₆It is selected from: the C of hydrogen, with or without substituent group_1-4Alkyl, OR₂₉Wherein R₂₉Selected from H, The C of with or without substituent group_1-6Alkyl.

In another preferred example, R₇It is selected from: the C of hydrogen, with or without substituent group_1-4Alkyl and NR₂₈R₂₉, wherein R₂₈, R₂₉It is independently selected from: the C of H or with or without substituent group_1-4Alkyl.

In another preferred example, R₈It is selected from: the C of hydrogen, halogen and with or without substituent group_1-4Alkyl.

In another preferred example, R₉It is selected from: the C of hydrogen, halogen and with or without substituent group_1-4Alkyl.

In another preferred example, R₁₀Be selected from: hydrogen, halogen, cyano, nitro and with or without substituent group C_1-4Alkane Base.

In another preferred example, R₄Selected from H, cyano, with or without substituent group C_1-6Alkyl.

The second aspect of the present invention, provides a kind of pharmaceutical composition, and the composition contains change described in first aspect Close object or its pharmaceutically acceptable salt, prodrug and pharmaceutically acceptable carrier.

In another preferred example, the effective quantity refers to therapeutically effective amount or inhibits effective quantity, preferably 0.01~ 99.99%.

In another preferred example, described pharmaceutical composition also includes another or a variety of antitumor agents.

In another preferred example, the pharmaceutical composition inhibits or degrades JAK1 and/or JAK2.

In another preferred example, the pharmaceutical composition is related for treating JAK1 and/or JAK2 activity or expression quantity Disease.

The third aspect of the present invention provides the purposes of compound as described in the first aspect of the invention, is used for:

(a) drug of preparation treatment disease relevant to JAK1 and/or JAK2 activity or expression quantity；

(b) agent of JAK1 and/or JAK2 targeted inhibition or degradation agent are prepared；

(c) inhibit to external non-therapeutic or the JAK1 and/or JAK2 that degrades；

(d) inhibit tumor cell proliferation to external non-therapeutic；And/or

(e) treatment disease relevant to JAK1 and/or JAK2 activity or expression quantity.

In another preferred example, it is described to JAK1 and/or JAK2 activity or the relevant disease of expression quantity be tumour or itself Immunological diseases.

In another preferred example, the disease relevant to JAK1 and/or JAK2 activity or expression quantity is selected from the group: organ Graft rejection, xenograft rejection, lupus erythematosus, multiple sclerosis, rheumatoid arthritis, psoriasis (psoriasis), cancer, Asthma, atopic dermatitis, type-1 diabetes mellitus and diabetic complication, autoimmune thyroid disorders, ulcerative colitis, gram Engler sieve disease, Alzheimer disease, leukaemia, lymthoma, Huppert's disease, alopecia areata and leucoderma etc..

The fourth aspect of the present invention provides a kind of preparation method of compound of formula I as described in the first aspect of the invention, Comprising steps of

(a) it in atent solvent, is reacted with formula IV compound and Formula II compound, obtains compound of formula I；

In the above formulas, each group is as defined above, M₁For leaving group.

In another preferred example, the method also includes steps:

(b) it in atent solvent, is reacted with formula III compound and Formula V compound, obtains Formula II compound, M₁、M₂For from Remove group.

The fifth aspect of the present invention provides a kind of inhibition or the method for the JAK1 and/or JAK2 that degrades, comprising steps of right Effective object applies a effective amount of compound of formula I or its pharmaceutically acceptable salt or right as described in the first aspect of the invention Object application is inhibited to inhibit a effective amount of pharmaceutical composition as described in the fourth aspect of the present invention.

In another preferred example, the inhibition or degradation are external non-therapeutics.

In another preferred example, when applying a effective amount of Formulas I chemical combination as described in the first aspect of the invention to effective object When object or its pharmaceutically acceptable salt, the inhibition effective quantity is 0.001-500nmol/L, preferably 0.01- 200nmol/L。

The sixth aspect of the present invention provides that a kind for the treatment of and JAK1 and/or JAK2 are active or the relevant disease of expression quantity Method, which comprises to treatment object application therapeutically effective amount Formulas I chemical combination as described in the first aspect of the invention Object, or the pharmaceutical composition as described in the 4th invention of the invention.

In another preferred example, the object is mammal；Preferably, the mammal is behaved.

It should be understood that above-mentioned each technical characteristic of the invention and having in below (eg embodiment) within the scope of the present invention It can be combined with each other between each technical characteristic of body description, to form a new or preferred technical solution.As space is limited, exist This no longer tires out one by one states.

Specific embodiment

The present inventor after extensive and in-depth study, is prepared for a kind of compound with structure shown in Formulas I, and find It inhibits with JAK1 and/or JAK2 and degrading activity.And the compound is at much lower concentrations, can to JAK1 and/or JAK2 generates inhibition and degradation, and activity is quite excellent, thus can be used for treatment and JAK1 and/or JAK2 activity or express Measure relevant disease such as tumour.The present invention is completed on this basis.

The present invention provides the purposes of a kind of noval chemical compound and its degrade tyrosine protein kinase JAK1 and/or JAK2.This The compound of invention is able to suppress or degrades JAK1 and/or JAK2, and can be used as JAK1 and/or JAK2 inhibitor or degradation agent And wave immunoregulation effect, and rheumatoid arthritis for treating mammal, psoriasis, psoriasis arthropathica, The diseases such as ankylosing spondylitis, atopic dermatitis, keratoconjunctivitis sicca and Crohn disease.

Term

In the present invention, term " C_1-8Alkyl " refers to the functional group for containing only two kinds of carbon, hydrogen atoms, wherein of carbon atom Number is 1~8.Alkyl can be regarded as corresponding hydrocarbon and lose remaining free radical after a hydrogen atom, can be alkyl, cycloalkanes Base, alkenyl or alkynyl etc.；Its structure can be straight chain, branch or ring-type；It can be aliphatic, be also possible to aromatic.

Term " C_1-6Alkyl " refers to the linear or branched alkyl group with 1~6 carbon atom, such as methyl, ethyl, propyl, different Propyl, butyl, isobutyl group, sec-butyl, tert-butyl, or similar group.

The term as used herein " alkoxy " includes O- alkyl, and " alkyl " therein is as defined above.

Term used herein " halogenated " unless otherwise directed, including fluoro, chloro, bromo or iodo.

The compound of the present invention can contain double bond.When containing this kind of double bond, the compound of the present invention is with cis-, trans- Or mixtures thereof exist.

Halogen described herein includes fluorine, chlorine, bromine and iodine.

Unless otherwise directed, the moieties of alkyl and alkoxy referred to herein can be straight chain, branch or ring-type 's.

In the present invention, term " cyclic hydrocarbon radical " refers to the functional group containing two kinds of carbon, hydrogen atoms.Including naphthenic base, cycloalkenyl (at least containing a carbon-carbon double bond) and aryl.They can be monocycle, bicyclic and polycyclic.They can be loop coil, can also be with It is condensed ring.

In the present invention, term " heterocyclic hydrocarbyl " refers to containing carbon, hydrogen and at least one heteroatomic function in addition to carbon, hydrogen Group.Including Heterocyclylalkyl, heterocycloalkenyl (at least containing a carbon-carbon double bond) and heteroaryl.One or more cyclization in ring is former Son is hetero atom.Hetero atom can be O, N and S atom and their various combinations.They can be monocycle, bicyclic and more Ring.They can be loop coil, be also possible to condensed ring.

In the present invention, term " substituent group " includes but is not limited to fluorine, chlorine, bromine, cyano, hydroxyl, amino, C_1-6Oxyl, C_1-6Halohydrocarbyl, C_1-6Acyl group, C_1-6Sulfonyl.

The term as used herein " oxyl " refers to O- alkyl, and " alkyl " therein is as defined above.

The term as used herein " hydrocarbon carbonyl oxygen " refers to C (=O) O- alkyl, and " alkyl " therein is as defined above.

The term as used herein " amido " refers to N (H or alkyl 1) (H or alkyl 2), and " alkyl " therein is as above to determine Justice.

The term as used herein " amino-carbonyl " refers to C (=O)-amido, and " amido " therein is as defined above.

The term as used herein " amide groups " refers to N (H or alkyl)-C (=O)-alkyl, and " alkyl " therein is institute as above Definition.

In the present invention, term " containing ", "comprising" or " comprising " indicate that various composition can be applied to of the invention mix together It closes in object or composition.Therefore, term " mainly by ... form " and " consist of " were included in term " containing ".

In the present invention, term " pharmaceutically acceptable " ingredient refers to suitable for people and/or animal and without excessive bad pair It reacts (such as toxicity, stimulation and allergy), that is, has the substance of reasonable benefit/risk ratio.

In the present invention, amount or table that term " effective quantity " refers to therapeutic agent treatment, alleviates or prevent target disease or situation Reveal the detectable amount for treating or preventing effect.The figure of the object is depended on for the accurate effective quantity of certain an object and is good for The combination of therapeutic agent and/or therapeutic agent that health situation, the property and degree of illness and selection are given.Therefore, standard is preassigned True effective quantity is useless.However, can determine the effective quantity with routine experiment for the situation that Mr. Yu gives, face Bed doctor can judge.

Herein, except place is illustrated, term " substitution " refers to that one or more hydrogen atoms on group are selected from down The substituent group of group replaces: halogen, unsubstituted or halogenated C_1-6Alkyl, unsubstituted or halogenated C_2-6It is acyl group, unsubstituted or halogenated C_1-6Alkyl-hydroxyl.

Unless stated otherwise, in the present invention, the compound occurred is intended to including all possible optical isomer, Such as the compound of single chiral or the mixture (i.e. racemic modification) of various different chipal compounds.All chemical combination of the invention Among object, each asymmetric carbon atom can be optionally the mixture of R configuration or S configuration or R configuration and S configuration.

As used herein, term " the compounds of this invention " refers to Formulas I compound represented.The term further includes and Formulas I chemical combination Various crystalline forms, pharmaceutically acceptable salt, hydrate or the solvate of object.

As used herein, term " pharmaceutically acceptable salt " refers to that the compounds of this invention and acid or alkali are formed by suitable use Make the salt of drug.Pharmaceutically acceptable salt includes inorganic salts and organic salt.A kind of preferred salt is the compounds of this invention and acid The salt of formation.The acid for suitably forming salt includes but is not limited to: hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid etc. are inorganic Acid, formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, Picric acid, methanesulfonic acid, benzene methanesulfonic acid, the organic acids such as benzene sulfonic acid；And the acidic amino acids such as aspartic acid, glutamic acid.

Compound and its pharmaceutically acceptable salt

The present invention relates to compounds of Formula I or its pharmaceutically acceptable salts；

Wherein:

Indicate singly-bound；

Indicate singly-bound or double bond；

W missing is selected from O, NR17 ,-X2C (=O) X3 ,-X2S (=O)_gX3；Wherein R17 is H, with or without takes The C of Dai Ji_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocycle Alkyl；Wherein X2, X3 are independent lacks or is selected from O, S, NR₁₈；The wherein integer that g is 0 to 2；Wherein R₁₈For H, have or Without the C of substituent group_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl；

A is the integer (such as 1,2,3,4,5) between 0 to 5；

B is the integer (such as 1,2,3) between 0 to 3；

C is the integer (such as 1,2,3,4,5,6,7,8,9) between 0 to 30；

D is the integer (such as 1,2,3,4,5) between 0 to 5

E is the integer (such as 0,1,2) between 0 to 2.

Unless otherwise instructed, the integer of this paper is 0,1,2,3,4,5,6,7,8 or 9.

It is preferably carried out in mode at of the invention one, the compound is selected from the group:

The compound of the present invention packet can form pharmaceutically acceptable salt with inorganic acid, organic acid or alkali.The nothing Machine acid includes but is not limited to hydrochloric acid, hydrobromic acid, nitric acid, perchloric acid, sulfuric acid or phosphoric acid etc.；The organic acid includes but is not limited to Methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, benzene sulfonic acid, p- toluenesulfonic acid, fumaric acid, oxalic acid, acetic acid, maleic acid, ascorbic acid, Lactic acid, tartaric acid, malonic acid, glycolic, succinic acid and propionic acid etc.；The alkali includes but is not limited to inorganic salts and amine.

Term pharmaceutically acceptable salt refers to according to medical judgment suitable for contacting the tissue of people and mammal and without mistake Spend those of toxicity, stimulation, allergic reaction etc. salt.Pharmaceutically acceptable salt is well known in the art.

Present invention also contemplates that the pharmaceutical composition of pro-drug containing a compound of formula I.Pro-drug includes suchization Object is closed, wherein precursor molecule passes through carbonic acid ester bond, urethane bond, amido bond, alkyl ester bond, phosphoric acid ester bond, phosphoramidic acid Ester bond is covalently bound on free carboxy, hydroxyl, amino or the amido of compound of formula I.

The preparation of compound

Preparation method

The preparation method of formula I structural compounds is described more particularly below, but these specific methods are not to this hair Bright composition any restrictions.The compounds of this invention can also be optionally by various conjunctions describing in the present specification or known in the art It combines at method and is easily made, such combination can readily be carried out by those skilled in the art in the invention.

Following reaction process illustrates the preparation of the compounds of this invention.Unless otherwise directed, reaction process and the discussion below In A, B, W, Y, Z, X, a, b, c, d, e, R₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉、R₁₀It is as defined above.

In general, the slave formula III as described in following proposal obtains the compound of Formulas I:

In compound II, when W is ether, formula III (W1=OH) can be used under alkali effect and containing leaving group Intermediate direct nucleophilic displacement of fluorine preparation, or reaction prolonged using light react with alcohol and prepare；When W is ester, carbamate (NHCO₂) When, formula III (W1=OH) can be used and react preparation with acyl chlorides, Acibenzolar (amide), carboxylic acid, isocyanates under alkali effect；Work as W When for amine, formula III (W1=NH can be used₂) prepared under alkali effect with the direct nucleophilic displacement of fluorine of the intermediate containing leaving group, Formula III (W1=NH can also be used₂) with aldehyde/ketone reductive amination process is carried out to prepare；When W is amide, alkoxy carbonyl group amine (OCONH) and when urea formula III (W1=NH, can be used₂) alkali effect under with corresponding acyl chlorides, Acibenzolar (amide), carboxylic acid, isocyanide Acid esters reaction preparation.

It, can be with directly replacing or prepared by reduction amination when A and nitrogen-atoms are keyed with C-N in compound I；When A with When nitrogen-atoms is connected in a manner of amide, urea, carbamate, sulfonamide, sulphamide, corresponding acyl chlorides, Acibenzolar (acyl can be used Amine), carboxylic acid, isocyanates, sulfonic acid chloride, chlorosulfuric acid preparation.

In general, the case where according to connection structure A and W, compound I can also connect formula IV and intermediate chain by elder generation It connects, then reacts to obtain with formula III.Chemical synthesis process used is same as above.

Formula III, IV compound can be obtained by known synthetic method or be easy through commercially available acquisition.

The application of compound of formula I

The compound of formula I can be used for one or more purposes below:

(d) inhibit tumor cell proliferation to external non-therapeutic；And/or

(e) treatment disease (such as autoimmune disease) relevant to JAK1 and/or JAK2 activity or expression quantity.

In another preferred example, the disease relevant to JAK1 and/or JAK2 activity or expression quantity is organ transplant row Reprimand, heterograft, lupus erythematosus, multiple sclerosis, rheumatoid arthritis, psoriasis (psoriasis), cancer, asthma, spy answer Property dermatitis, type-1 diabetes mellitus and diabetic complication, autoimmune thyroid disorders, ulcerative colitis, Crohn's disease, Alzheimer disease, leukaemia, lymthoma, Huppert's disease, alopecia areata, leucoderma etc..

Compound of formula I of the invention can be used for preparing a kind of pharmaceutical composition, and the pharmaceutical composition includes: that (i) has The compound of formula I of effect amount or its pharmaceutically acceptable salt；(ii) pharmaceutically acceptable carrier.

In another preferred example, the effective quantity refers to therapeutically effective amount or inhibits effective quantity.

The method that compound of formula I of the invention can be also used for inhibition or degrade cloth JAK1 and/or JAK2, the inhibition It is that the inhibition of external non-therapeutic is also possible to therapeutic inhibition.

In another preferred example, when to inhibition object application a effective amount of compound of formula I of the invention of inhibition or its pharmacy When upper acceptable salt, the inhibition effective quantity is 0.001-500nmol/L, preferably 0.01-200nmol/L.

Particularly, the present invention also provides a kind for the treatment of diseases relevant to JAK1 and/or JAK2 activity or expression quantity Method, which comprises compound of formula I to treatment object application therapeutically effective amount described contains compound of formula I conduct The pharmaceutical composition of effective component.

Pharmaceutical composition and method of administration

Since the compounds of this invention can degrade JAK1 and/or JAK2 significantly to play and live to JAK1 and/or JAK2 inhibition Property, therefore the compounds of this invention and its various crystal forms, pharmaceutically acceptable inorganic or organic salt, hydrate or solvate, And containing the compounds of this invention be main active pharmaceutical composition can be used for treating, prevent and alleviate by with JAK1 And/or JAK2 activity or the relevant disease of expression quantity.According to the prior art, the compounds of this invention can be used for treating including tumour etc. Disease.

Pharmaceutical composition of the invention include safe and effective amount within the scope of the compounds of this invention or its be pharmacologically subjected to Salt and pharmacologically acceptable excipient or carrier.Wherein " safe and effective amount " refers to: the amount of compound is enough obviously Improve the state of an illness, and is unlikely to generate serious side effect.In general, pharmaceutical composition contains 1-2000mg the compounds of this invention/agent, More preferably, containing 5-200mg the compounds of this invention/agent.Preferably, described is " one " for a capsule or tablet.

" pharmaceutically acceptable carrier " refers to: one or more biocompatible solids or liquid filler or jello Matter, they are suitable for people's use and it is necessary to have enough purity and sufficiently low toxicity." compatibility " referred to herein as combines In object each component energy and the compound of the present invention and they between mutually admix, and significantly reduce the drug effect of compound.Medicine Acceptable carrier part example has cellulose and its derivates (such as sodium carboxymethylcellulose, ethyl cellulose sodium, fibre on Tie up plain acetic acid esters etc.), gelatin, talcum, solid lubricant (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (such as soya-bean oil, sesame Sesame oil, peanut oil, olive oil etc.), polyalcohol (such as propylene glycol, glycerol, mannitol, sorbierite), emulsifier Wetting agent (such as lauryl sodium sulfate), colorant, flavoring agent, stabilizer, antioxidant, preservative, apirogen water.

The method of application of the compounds of this invention or pharmaceutical composition is not particularly limited, and representative method of application includes (but being not limited to): in oral, tumor, rectum, parenteral (intravenous, intramuscular or subcutaneous) and local administration.

Solid dosage forms for oral administration includes capsule, tablet, pill, powder and granule.In these solid formulations In type, reactive compound is mixed at least one conventional inert excipients (or carrier), such as sodium citrate or Dicalcium Phosphate, or with Following compositions mixing: (a) filler or expanding material, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid；(b) it bonds Agent, for example, hydroxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and Arabic gum；(c) moisturizer, example Such as, glycerol；(d) disintegrating agent, for example, agar, calcium carbonate, potato starch or tapioca, alginic acid, certain composition silicates, And sodium carbonate；(e) retarding solvent, such as paraffin；(f) absorbsion accelerator, for example, quaternary ammonium compound；(g) wetting agent, such as spermaceti Pure and mild glycerin monostearate；(h) adsorbent, for example, kaolin；(i) lubricant, for example, talcum, calcium stearate, tristearin Or mixtures thereof sour magnesium, solid polyethylene glycol, lauryl sodium sulfate,.In capsule, tablet and pill, dosage form also may include Buffer.

Coating and shell material preparation can be used in solid dosage forms such as tablet, sugar-pill, capsule, pill and granule, such as casing and Other materials well known in the art.They may include opacifying agent, also, reactive compound or compound in this composition Release can discharge in certain a part in the digestive tract in a delayed fashion.The example of adoptable embedding component is polymeric material And wax material.When necessary, reactive compound can also be with one of above-mentioned excipient or a variety of formation microencapsulation forms.

Liquid formulation for oral administration includes pharmaceutically acceptable lotion, solution, suspension, syrup or tincture. In addition to active compounds, liquid dosage form may include the inert diluent routinely used in this field, such as water or other solvents, increase Solvent and emulsifier, example know, ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-BDO, dimethyl formyl The mixture of amine and oil, especially cottonseed oil, peanut oil, maize germ, olive oil, castor oil and sesame oil or these substances Deng.

Other than these inert diluents, composition also may include auxiliary agent, such as wetting agent, emulsifier and suspending agent, sweet taste Agent, tender taste agent and fragrance.

In addition to active compounds, suspension may include suspending agent, for example, ethoxylation isooctadecane alcohol, polyoxyethylene Sorbierite and Isosorbide Dinitrate, microcrystalline cellulose, aluminium methoxide and agar or the mixture of these substances etc..

Composition for parenteral injection may include physiologically acceptable sterile, aqueous or anhydrous solution, dispersion liquid, Suspension or lotion, and the aseptic powdery for re-dissolving into sterile Injectable solution or dispersion liquid.It is suitable aqueous and Nonaqueous carrier, diluent, solvent or excipient include water, ethyl alcohol, polyalcohol and its suitable mixture.

The dosage form of the compounds of this invention for local administration includes ointment, powder, patch, stock solution and inhalant. Active constituent aseptically with physiologically acceptable carrier and any preservative, buffer, or when necessary may need Propellant be mixed together.

The compounds of this invention can be administered alone, or be administered in combination with other pharmaceutically acceptable compounds.

It is the mammal that the compounds of this invention of safe and effective amount is applicable to treatment when using pharmaceutical composition (such as people), wherein dosage is the effective dosage pharmaceutically thought when application, for the people of 60kg weight, day is to medicament Amount is usually 1~2000mg, preferably 5~500mg.Certainly, specific dosage be also contemplated that administration route, patient health situation etc. because Element, within the scope of these are all skilled practitioners technical ability.

Main advantages of the present invention include:

1. providing a kind of compound shown in formula I.

2. JAK1 and/or JAK2 inhibitor and degradation agent and its preparation and the application of a kind of structure novel are provided, it is described Inhibitor can inhibit and degrade JAK1 and/or JAK2 at much lower concentrations.

3. providing the pharmaceutical composition of a kind for the treatment of and JAK1 and/or JAK2 activity related diseases.

Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention Rather than it limits the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually according to conventional strip Part, or according to the normal condition proposed by manufacturer.Unless otherwise stated, otherwise percentage and number are weight percent and weight Number.

The preparation of 1 compound 3 of embodiment

Step 1:

2.14g 6- bromine caproyl chloride and 1.37g pomalidomide are dissolved in 50ml THF (tetrahydrofuran), 8h is stirred at reflux, It is down to room temperature, is dried in vacuo to obtain compound (1) 2.19g for 40 DEG C after concentration.MS(ESI):450[M+H]⁺。

Step 2:

1g compound (1), 1.28g compound (2) and 2.6g DIPEA (n,N-diisopropylethylamine) are dissolved in 50ml In DMF (n,N-Dimethylformamide), 80 DEG C of stirring 6h are down to room temperature, chromatograph to obtain compound (3) 810mg through column after concentration, receive Rate: 65.6%.MS(ESI):649[M+H]⁺。¹H NMR (400MHz, CDCl3) δ 10.62 (br, 1H), 10.15 (br, 1H), 9.02 (s, 1H), 8.46 (s, 1H), 8.24 (s, 1H), 7.45 (t, J=7.6Hz, 1H), 7.35 (dd, J=3.2,2.0Hz, 1H), 7.11 (d, J=7.6Hz, 1H), 6.88 (d, J=7.6Hz, 1H), 6.70 (dd, J=3.6,1.6Hz, 1H), 6.45 (t, J =4.0Hz, 1H), 4.94 (dd, J=12.0,5.6Hz, 1H), 3.65-3.85 (m, 4H), 3.40 (s, 2H), 2.71-2.88 (m, 4H), 1.83-2.52 (m, 6H), 1.60 (m, 2H), 1.41 (m, 2H).

The preparation of 2 compound 7 of embodiment

Step 1:

4- hydroxyl Thalidomide 100mg, triethylene glycol list benzyl oxide 96mg, triphenylphosphine 100mg are dissolved in the anhydrous THF of 10ml In, DIAD (diisopropyl azodiformate) 95mg is added dropwise, reacts at room temperature 2h.Decompression removes THF, obtains chemical combination after column chromatographic purifying Object (4) 110mg.MS(ESI):497[M+H]⁺。

Step 2:

100mg compound (4) and 10%Pd-C 100mg are added in methanol 10ml, room temperature hydrogenated over night.Filtering, filtrate Concentration, residue obtain compound (5) 40mg after column chromatographic purifying.MS(ESI):407[M+H]⁺。

Step 3:

Compound (5) 30mg is dissolved in methylene chloride 5ml, Dess-martin oxidant 47mg, room temperature reaction is added 3h.NaHCO is added₃Saturated aqueous solution and Na₂S₂O₃Saturated aqueous solution stirs 5min.Organic layer is separated, through anhydrous Na₂S₂O₃It is dry It is dry, it is concentrated to dryness, obtains compound (6) and be directly used in the next step.

After compound (6) are dissolved with methylene chloride 7ml, it is added raw material (2) 45mg and NaBH (OAc)₃23mg, room temperature Reaction is overnight.It removes methylene chloride under reduced pressure, obtains compound (7) after column chromatographic purifying, 25mg.MS (ESI): 668 [M+H]⁺。¹H NMR(400MHz,CDCl3)δ10.69(br,1H),10.26(br,1H),8.78(s,1H),8.44(s,1H),8.28(s,1H), 7.52 (t, J=8.0Hz, 1H), 7.30 (m, 2H), 7.10 (d, J=8.4Hz, 1H), 6.71 (d, J=3.2Hz, 1H), 5.06 (m, 1H), 4.31 (m, 2H), 3.88-4.04 (m, 2H), 3.73-3.84 (m, 6H), 3.67 (br, 2H), 3.58 (t, J= 4.8Hz,2H),3.41(s,2H),2.75-2.89(m,4H),2.06-2.35(m,2H)。

The preparation of 3 compound 10 of embodiment

Step 1:

By 2- (2- (2- bromine oxethyl) ethyoxyl) benzyl acetate 174mg, potassium carbonate 100mg, potassium iodide 20mg and pool horse It spends amine 100mg to be added in 20mlDMF (n,N-Dimethylformamide), 80 DEG C of reactions are overnight.Reaction solution obtains after column chromatographic purifying Compound (8) 113mg.MS(ESI):510[M+H]⁺。

Step 2:

100mg compound (8) and 10%Pd-C100mg are added in 10ml methanol, room temperature hydrogenated over night.Filtering, filtrate Concentration, residue obtain compound (9) 73mg after column chromatographic purifying.MS (ESI anion): 418 [M-H]^-。

Step 3:

Compound (9) 50mg and raw material (2) 50mg are dissolved in methylene chloride 5ml, HOBt (1- hydroxy benzo three is added Azoles) 20mg and EDC (1- (3- dimethylamino-propyl) -3- ethyl-carbodiimide hydrochloride) 40mg, room temperature reaction is overnight.Decompression is steamed Except methylene chloride, compound (10) are obtained after column chromatographic purifying, 23mg.MS (ESI): 681 [M+H]⁺。¹H NMR (400MHz, CDCl3) δ 10.56 (br, 1H), 10.08 (br, 1H), 8.81 (s, 1H), 8.46 (s, 1H), 8.28 (s, 1H), 7.42 (dd, J= 8.0,7.2Hz, 1H), 7.33 (dd, J=3.2,2.0Hz, 1H), 7.03 (d, J=7.2Hz, 1H), 6.84 (d, J=8.8Hz, 1H), 6.71 (dd, J=3.6,1.6Hz, 1H), 6.46 (t, J=5.2Hz, 1H), 4.94 (dd, J=12.0,5.2Hz, 1H), 4.51 (s, 2H), 3.65-3.75 (m, 4H), 3.57 (t, J=4.8Hz, 2H), 3.45 (m, 4H), 3.40 (s, 2H), 2.71- 2.88 (m, 4H), 2.1 (m, 2H).

The preparation of 4 compound 13 of embodiment

Step 1:

Raw material (2) 200mg is dissolved in methylene chloride 10ml, triethylamine 100mg and 5- benzyloxy pentane sulfonic acid chloride is added 200mg, room temperature reaction overnight, remove methylene chloride under reduced pressure, obtain compound (11) 312mg after column chromatographic purifying.MS(ESI):520 [M+H]⁺。

Step 2:

300mg compound (11) and 300mg 10%Pd-C are added in methanol 20ml, room temperature hydrogenation is stayed overnight.It crosses Filter, filtrate are concentrated, and obtain compound (12) 211mg.MS (ESI): 430 [M+H] after column chromatographic purifying⁺。

Step 3:

4- hydroxyl Thalidomide 100mg, triphenylphosphine 100mg and compound (12) 150mg are dissolved in 30ml anhydrous four In hydrogen furans, DIAD (diisopropyl azodiformate) 100mg is added dropwise, reacts at room temperature 3h, decompression removal tetrahydrofuran, column chromatography Compound (13) 97mg is obtained after purification.MS(ESI):686[M+H]⁺。¹H NMR(400MHz,CDCl3)δ10.55(br,1H), 10.17 (br, 1H), 8.80 (s, 1H), 8.41 (s, 1H), 8.29 (s, 1H), 7.53 (t, J=7.6Hz, 1H), 7.31 (m, 2H), 7.08 (d, J=8.0Hz, 1H), 6.75 (d, J=3.6Hz, 1H), 5.01 (m, 1H), 3.75-3.95 (m, 6H), 3.53 (m, 2H),3.38(s,2H),2.58-2.77(m,2H),2.06-2.35(m,2H),1.63-1.95(m,4H),1.40(m,2H)。

It see the table below by compound prepared by the method for similar embodiment 2,3:

It see the table below by compound prepared by the method for similar embodiment 2:

Embodiment 5 tests compound to the inhibitory activity of JAK1/2 with Caliperassay method

Experimental procedure is as follows:

Configure 1 × kinase reaction buffer (50mM HEPES, PH 7.5；0.0015%Brij-35) and kinase reaction is whole Only liquid (100mM HEPES, PH 7.5；0.0015%Brij-35；0.2%Coating Reagent；50mM EDTA)；

Configuration laboratory sample: the sample solution (100%DMSO dissolution) of 5 μM of 100 μ L is added in 96 orifice plates, obtains 50 × sample solution.Two are arranged on same plate and contains only the hole of 100 μ L 100%DMSO as control.One conduct is not added Sample controls, another is used as not enzyme control.10 μ L samples and 90 μ L 1 × kinase reaction buffers are added in 96 orifice plates, As transit plate.This transit plate is shaken 10 minutes.

Prepare breadboard: taking in 96 hole transit plates configured each 5 μ L of sample into 384 orifice plates.

Kinase reaction: the 2.5 of 10 μ L are added in 55 × compound solutions of μ L (being dissolved with DMSO, be diluted with water 10 times) × JAK1/2 kinase solution (kinases with 1 × kinase reaction buffer dilute), add 10 μ L's after being incubated at room temperature 10min 2.5 × substrate peptide solution (in the diluted FAM label peptide of 1 × kinase reaction buffer and ATP).

It terminates kinase reaction: 25 μ L kinase reaction terminate liquids is added after reacting a period of time at 28 DEG C.

Fluorescence (F) is tested on Caliper, and collects data.

Calculate kinase activity inhibiting rate: to percent inhibition=(F of kinase activity_{DMSO control}-F_Sample)/(F_{DMSO control}-F_{Negative control}) × 100, using DMSO as solution control, kinases is not added as negative control.

The results show that compound (concentration 100nM) see the table below the inhibitory activity of JAK1/2:

The activity of the Western blot detection compound degradation JAK1 and JAK2 albumen of embodiment 6

Cell strain: Jurkat cell strain uses the RPMI1640 culture containing 10% calf serum to be based on 37 DEG C, 5%CO₂, saturation Culture in humidified incubator.

DMSO control group, compound intervention group (10 μM) are set, handles and 100 μ L pre-cooling is added after collecting within 6 hours cell Cell pyrolysis liquid cracks 30min on ice, extracts total protein of cell, and bicinchoninic acid (BCA) method measurement protein concentration is simultaneously determined Amount.Conventional glue, loading, electrophoresis, then transferring film, closing, be separately added into rabbit-anti people JAK1 (1: 500) and rabbit-anti people JAK2 (1: 500) goat anti-rabbit igg (1: 5000) of horseradish peroxidase-labeled, ECL after rinsing is added in, 4 DEG C of overnight incubations after rinsing Developer solution colour developing, Bio-Rad gel imaging system scanning imagery, software processing analysis.With glyceraldehyde phosphate dehydrogenase (GAPDH) internal reference compares.

Gray analysis is carried out to each band using Image J software, calculates the drop of degradation JAK1 or JAK2 albumen Solution rate.Calculation formula is as follows:

The results show that compound (10 μM) see the table below the degradation rate of JAK1/2 albumen in Jurkat cell:

7 CCK8 method of embodiment detects formula compound and makees to the inhibition that Jurkat cell (human leukemia T lymphocyte) is proliferated With

In vitro with CCK8 method measuring compound to the inhibited proliferation of Jurkat cell.Specific step is as follows:

It takes Jurkat cell to be incubated in 1640 culture mediums of 10% calf serum, is inoculated in 96 orifice plates, 2 × 10⁵Cell/ Hole is placed in 37 DEG C, 5%CO₂In incubator.Compound is dissolved in dimethyl sulfoxide (DMSO), obtaining concentration is the molten of 10mM Liquid, then it is diluted to required concentration with phosphate buffer, it is separately added into above-mentioned 96 orifice plate, each 2 hole of concentration, every 10 μ l of hole, often A concentration makees two parallel testings.It is added in plate after DMSO accordingly to be made to gradient dilution, as control.

By above-mentioned 96 orifice plate in 37 DEG C, 5%CO₂After cultivating 48 hours in cell incubator, 10 μ lCCK8 are added in every hole Solution continues to keep the temperature 1~4 hour in the incubator.Measure 465nm absorbance value.

Comparative survival rate of cells after compound is handled is calculated according to absorbance value.Calculation formula is as follows:

Compound is calculated to the IC of Jurkat cell by software₅₀。

The results show that the compound of synthesis see the table below the in-vitro multiplication inhibiting effect of Jurkat cell:

The above result shows that the compound of synthesis has the good activity for inhibiting Jurkat cell.

Resisting rheumatoid arthritis activity in 8 test body of embodiment

Using full Freund's adjuvant induction Wistar rat rheumatoid arthritis as model, the anti-class wind of compound (3) is measured Wet arthritis effect.Specific step is as follows:

1) test specimen

Sample: compound (3).

2) preparation method

Sample: compound (3), when preparation, are dissolved with normal saline solution

3) animal and adjuvant

Wistar rat 18, male, weight 170-200g.

Adjuvant: full Freund's adjuvant (CFA), Sigma.

4) test method

Wistar rat is divided into 3 groups, every group 6 at random.Respectively blank control group, model group, (compound (3) Group (20mg/kg ip).Rheumatoid arthritis is induced in the left back vola pedis intracutaneous injection CFA 0.1mL of rat.The 11st after rat modeling It starts drug treatment, once a day, the 22nd day treatment end.Left back sufficient volume is measured after treatment end, calculates foot swelling suppression Rate processed.

5) test result

Compound (3) see the table below the foot swelling inhibiting rate of full Freund's adjuvant induced rat rheumatoid arthritis:

Group	Foot swelling inhibiting rate (%)
		Blank control group	-
Model group	-
		Compound (3)	85

Above experiment in vivo is the result shows that compound (3) has good resisting rheumatoid arthritis activity in vivo.

All references mentioned in the present invention is incorporated herein by reference, independent just as each document It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can To make various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims It encloses.

Claims

1. a kind of such as following formula I compound represented or its pharmaceutically acceptable salt:

Wherein:

Indicate singly-bound；

Indicate singly-bound or double bond；

A missing is selected from C (=O), C (=O) X1, SOX1, SO₂The C of X1, with or without substituent group_1-8Alkyl has or not C with substituent group_1-8The C of cyclic hydrocarbon radical and with or without substituent group_1-8Heterocyclic hydrocarbyl；Wherein X1 is lacked or is selected from (CR₁₂R₁₃)_fO、(CR₁₂R₁₃)_fS and NR₁₄；Wherein R₁₂、R₁₃、R₁₄Independent is the C of H, with or without substituent group_1-8 Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical and with or without substituent group C_1-8Heterocyclic hydrocarbyl, f For the integer (such as 1,2 or 3) between 0 to 3；

W missing is selected from O, NR₁₇,-X2C (=O) X3 ,-X2S (=O)_gX3；Wherein R₁₇For H, with or without substituent group C_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl； Wherein X2, X3 are independent lacks or is selected from O, S, NR₁₈；The wherein integer that g is 0 to 2；Wherein R₁₈For H, with or without The C of substituted base_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8's Heterocyclic hydrocarbyl；

Y is (CR₂₂R₂₃)_h、CHX4(CR₂₂R₂₃)_h, CX4=CH (CR₂₂R₂₃)_hOr (CR₂₂R₂₃)_h；Wherein h is whole between 0 to 30 Number；Wherein R₂₂、R₂₃It is independently selected from the C of H, cyano, hydroxyl, amino, with or without substituent group_1-8Alkyl, band Have or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl, with or without The C of substituent group_1-8Oxyl；Wherein X4 is selected from H, halogen, cyano, nitro, hydroxyl, with or without substituent group C_1-8Hydrocarbon oxygen Base, with or without substituent group C_1-8Hydrocarbon carbonyl oxygen, with or without substituent group C_1-8Amido, with or without substituent group C_1-8Ester group, with or without substituent group C_1-8Amino-carbonyl, with or without substituent group C_1-8Alkyl, with or without Substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl；

Z is (CR₂₄R₂₅)_i、CHX5(CR₂₄R₂₅)_i, CX5=CH (CR₂₄R₂₅)_iOr C ≡ C (CR₂₄R₂₅)_i；Wherein i is between 0 to 30 Integer；Wherein R₂₄、R₂₅It is independently selected from the C of H, cyano, hydroxyl, amino, with or without substituent group_1-8Hydrocarbon Base, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl, with or without substituted Base C_1-8Oxyl；Wherein X5 is selected from H, halogen, cyano, nitro, hydroxyl, with or without substituent group C_1-8Oxyl, band Have or without substituent group C_1-8Hydrocarbon carbonyl oxygen, with or without substituent group C_1-8Amido, with or without substituent group C_1-8Ester Base, with or without substituent group C_1-8Amino-carbonyl, with or without substituent group C_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8Heterocyclic hydrocarbyl；

B missing is selected from O, C=O, S, NR₁₅、-NR₁₅C (=O)-,-C (=O) NR₁₅,-C (=O) O-, OC (=O) O- ,- NR₁₅C (=O) O- ,-OC (=O) NR₁₅-、-NR₁₅C (=O) NR₁₆, with or without substituent group C_1-12Alkyl, have Or the C without substituent group_1-12Cyclic hydrocarbon radical and with or without substituent group C_1-12Heterocyclic hydrocarbyl；Wherein R₁₅、R₁₆Respectively From the independent C selected from H, with or without substituent group_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, With the C of with or without substituent group_1-8Heterocyclic hydrocarbyl；

X is selected from CR₁₉R₂₀, C (=O), S (=O), SO₂、NR₂₁；Wherein R₁₉、R₂₀It is independently selected from H, cyano, hydroxyl, ammonia The C of base, with or without substituent group_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without The C of substituent group_1-8Heterocyclic hydrocarbyl, with or without substituent group C_1-8Oxyl；Wherein R₂₁Selected from H, with or without The C of substituted base_1-8Alkyl, with or without substituent group C_1-8Cyclic hydrocarbon radical, with or without substituent group C_1-8's Heterocyclic hydrocarbyl；

R₁、R₈It is each independently selected from the C of H, with or without substituent group_1-8The cyclic hydrocarbon of alkyl, with or without substituent group Base, the heterocyclic hydrocarbyl of with or without substituent group, with or without substituent group C_1-6Acyl group；

R₂、R₅It is independently selected from: hydrogen, OR₃₃、NR₃₄R₃₅, cyano, halogen, with or without substituent group C_1~8Alkyl, band Have or without the cyclic hydrocarbon radical of substituent group, the heterocyclic hydrocarbyl of with or without substituent group, with or without substituent group C_1-6 The amide groups of acyl group, with or without substituent group；Wherein R₃₃、R₃₄、R₃₅It is independently selected from H, with or without substituted The C of base_1~8Alkyl, with or without substituent group cyclic hydrocarbon radical, the heterocyclic hydrocarbyl of with or without substituent group；

R₃、R₆、R₇、R₉、R₁₀It is independently selected from: H, OR₂₇、NR₂₈R₂₉, cyano, halogen, nitro, with or without substituted The C of base_1-8Alkyl, the cyclic hydrocarbon radical of with or without substituent group, the heterocyclic hydrocarbyl of with or without substituent group, X6S (=O)_kR₃₀, X6C (=O) R₃₁；Wherein k is the integer between 0 to 2；Wherein R₂₇, R₂₈, R₂₉, R₃₀, R₃₁It is independently selected from H, has Or the C without substituent group_1~8Alkyl, cyclic hydrocarbon radical, heterocyclic hydrocarbyl；Wherein X6 lacks or is selected from O, S, NR₃₂；Wherein R₃₂For H, band Have or without substituent group C_1-8Alkyl, with or without substituent group cyclic hydrocarbon radical and with or without substituent group Heterocyclic hydrocarbyl；

R₄Selected from H, cyano, carboxyl, with or without substituent group C_1-8Alkyl, with or without substituent group hydrocarbon oxygen carbonyl Base；

A is the integer (such as 1,2,3,4,5) between 0 to 5；

B is the integer (such as 1,2,3) between 0 to 3；

C is the integer between 0 to 30；

D is the integer between 0 to 5；

E is the integer between 0 to 2.

2. compound as described in claim 1 or its pharmaceutically acceptable salt, it is characterised in that:

In the Formulas I, A missing；W is X2C (=O) X3, wherein X3 NR₁₈And X2 missing or X2 are NR₁₈And X3 Missing；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4Hydrocarbon Base, h are the integer between 1 to 6；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from H, hydroxyl, with or without The C of substituent group_1-4Alkyl, i be 1 to 6 between integer；C is 0；Or

In the Formulas I, A missing；W missing is O；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from H, hydroxyl, band Have or without substituent group C_1-4Alkyl, h be 0 to 3 between integer；B is O；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅Respectively The independent C for being selected from H, hydroxyl, with or without substituent group_1-4Alkyl, i be 0 to 3 between integer；C is between 1 to 6 Integer；Or

In the Formulas I, A is C (=O) X1, and wherein X1 lacks or be selected from (CR₁₂R₁₃)_fAnd (CR O,₁₂R₁₃)_fS, wherein f be 0 to 2 it Between integer, R₁₂、R₁₃、R₁₄Independent is hydrogen or C_1-4Alkyl；W is NR₁₇, wherein R₁₇For H or with or without The C of substituent group_1-4Alkyl；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from H, hydroxyl, with or without substituted The C of base_1-4Alkyl, h be 0 to 3 between integer；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from H, hydroxyl, band Have or without substituent group C_1-4Alkyl, i be 0 to 3 between integer；B is O；C is the integer between 1 to 4；Or

In the Formulas I, A SO₂X1, wherein X1 lacks or is selected from O and S；W is O；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃Respectively solely The vertical C for being selected from H, hydroxyl, with or without substituent group_1-4Alkyl, h be 1 to 6 between integer；Z is (CR₂₄R₂₅)_i, Middle R₂₄、R₂₅It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4Alkyl, i be 0 to 3 between integer； C is 0；Or

In the Formulas I, A missing；W is NR₁₇, wherein R₁₇For the H or C of with or without substituent group_1-4Alkyl；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4Alkyl, h be 0 to 3 Between integer；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from the C of H, hydroxyl, with or without substituent group_1-4 Alkyl, i be 0 to 4 between integer；B is O；C is the integer between 1 to 6；Or

In the Formulas I, A missing；W missing；Y is (CR₂₂R₂₃)_h, wherein R₂₂、R₂₃It is independently selected from H, hydroxyl, has or not C with substituent group_1-4Alkyl, h be 0 to 3 between integer；Z is (CR₂₄R₂₅)_i, wherein R₂₄、R₂₅It is independently selected from H, the C of hydroxyl, with or without substituent group_1-4Alkyl, i be 0 to 3 between integer；B is O；C be 1 to 10 between (preferably Between 1 to 6) integer.

3. compound as described in claim 1 or its pharmaceutically acceptable salt, which is characterized in that in Formulas I, X be C (= O)。

4. compound as described in claim 1 or its pharmaceutically acceptable salt, which is characterized in that R₁It is selected from: H and having Or the C without substituent group_1-4Alkyl.

5. compound as described in claim 1 or its pharmaceutically acceptable salt, which is characterized in that R₄Selected from H, cyano, band Have or without substituent group C_1-6Alkyl.

6. compound as described in claim 1 or its pharmaceutically acceptable salt, which is characterized in that the compound is selected from The following group:

7. a kind of pharmaceutical composition, which is characterized in that the composition contains compound described in claim 1 or its pharmacy Upper acceptable salt, prodrug and pharmaceutically acceptable carrier.

8. the purposes of a kind of compound as described in claim 1 or its pharmaceutically acceptable salt, is used for:

(d) inhibit tumor cell proliferation to external non-therapeutic；And/or

9. purposes as claimed in claim 8, which is characterized in that described relevant to JAK1 and/or JAK2 activity or expression quantity Disease is tumour or autoimmune disease.

10. the method for a kind of inhibition or degradation JAK1 and/or JAK2, which is characterized in that comprising steps of having to effective object application The compound of formula I as described in claim 1 or its pharmaceutically acceptable salt of effect amount, or inhibit effective to inhibiting object to apply The pharmaceutical composition as claimed in claim 7 of amount.