CN109963853B

CN109963853B - Compound with activity of degrading tyrosine protein kinase JAK3

Info

Publication number: CN109963853B
Application number: CN201880004075.3A
Authority: CN
Inventors: 舒永志
Original assignee: Shanghai Meizer Pharmaceuticals Co ltd
Current assignee: Shanghai Meizer Pharmaceuticals Co ltd
Priority date: 2017-09-03
Filing date: 2018-09-02
Publication date: 2022-04-22
Anticipated expiration: 2038-09-02
Also published as: WO2019042443A1; CN109422751B; CN109963853A; CN109422751A

Abstract

A compound with the activity of degrading tyrosine protein kinase JAK3 has a structural formula shown in a formula I, and can be used for preparing medicines for treating JAK3 related diseases.

Description

Compound with activity of degrading tyrosine protein kinase JAK3

Technical Field

The invention belongs to the field of medicines, and particularly relates to a compound with the activity of degrading tyrosine protein kinase JAK3, and preparation and application thereof.

Background

Janus kinases (JAKs) are non-receptor tyrosine protein kinases existing in cells, are very important for multiple functions such as growth, activation and homeostatic regulation of T cells, and play a key role in regulating the cell functions of the lymphohematopoietic system. In mammals, this family contains 4 major family members, namely JAK1, JAK2, JAK3 and Tyk 2. Among them, JAK1, JAK2 and Tyk2 are widely expressed in various histiocytes, and JAK3 is mainly expressed in hematopoietic tissues at high efficiency, such as activated B lymphocytes, T lymphocytes, bone marrow cells and thymocytes.

Due to the wide regulation of the JAK3-STAT signal pathway, organ transplant rejection, xenotransplantation, lupus erythematosus, multiple sclerosis, rheumatoid arthritis, psoriasis, cancer, asthma, atopic dermatitis, type I diabetes and diabetic complications, autoimmune thyroid disorders, ulcerative colitis, crohn's disease, alzheimer's disease, leukemia, lymphoma, multiple myeloma, alopecia areata, vitiligo and many other diseases occur, JAK3 is also involved, JAK3 plays a key role in disease occurrence as a signal transmission member, and becomes a drug target for developing and treating such diseases in medicine development.

Therefore, the skilled person is working to develop compounds capable of inhibiting JAK3 activity.

Disclosure of Invention

The invention aims to provide a compound capable of inhibiting and degrading JAK3, and preparation and application thereof.

In a first aspect of the invention, there is provided a compound of formula I, or a pharmaceutically acceptable salt thereof:

wherein:

represents a single bond;

represents a single bond or a double bond;

a is absent or selected from C (═ O), C (═ O) X1, SOX1, SO₂X1, with or without substituentsC of (A)_1-8Hydrocarbyl, C with or without substituents_1-8Cycloalkyl group, and C with or without substituent_1-8A heterocyclic hydrocarbon group; wherein X1 is absent or selected from (CR)₁₂R₁₃)_eO、(CR₁₂R₁₃)_eS and NR₁₄(ii) a Wherein R is₁₂、R₁₃、R₁₄Each independently selected from H, C with or without substituents_1-8With or without substituents C_1-8And C with or without substituents_1-8E is an integer between 0 and 3 (e.g., 1, 2, or 3);

w is absent or selected from O, NR₁₇、-X2C(＝O)X3、-X2S(＝O)_gX3; wherein R is₁₇Is H, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8The heterocyclic hydrocarbon group of (a); wherein X2 and X3 are independently deleted or selected from O, S, NR₁₈、(CH₂)_gO; wherein each g is an integer from 0 to 2; wherein R is₁₈Is H, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8The heterocyclic hydrocarbon group of (a);

y is (CR)₂₂R₂₃)_h、CHX4(CR₂₂R₂₃)_h、CX4＝CH(CR₂₂R₂₃)_hOr (CR)₂₂R₂₃)_h(ii) a Wherein h is an integer between 0 and 30; wherein R is₂₂、R₂₃Each independently selected from H, cyano, hydroxy, amino, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8A heterocyclic hydrocarbon group of (A), C with or without substituents_1-8Hydrocarbyloxy groups of (a); wherein X4 is H, halogen, cyano, nitro, hydroxy, with or without a substituent C_1-8Hydrocarbyloxy, with or without substituents C_1-8Alkoxycarbonyl, with or without substituents C_1-8Amino, with or without substituents C_1-8Ester group, with or without substituents C_1-8Aminocarbonyl, with or without substituents C_1-8Hydrocarbyl, with or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8A heterocyclic hydrocarbon group;

z is (CR)₂₄R₂₅)_i、CHX5(CR₂₄R₂₅)_i、CX5＝CH(CR₂₄R₂₅)_iOr C ≡ C (CR)₂₄R₂₅)_i(ii) a Wherein i is an integer between 0 and 30; wherein R is₂₄、R₂₅Each independently selected from H, cyano, hydroxy, amino, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8Heterocycloalkyl radicals, with or without substituents C_1-8A hydrocarbyloxy group; wherein X5 is H, halogen, cyano, nitro, hydroxy, with or without a substituent C_1-8With or without substituents C_1-8Alkoxycarbonyl, with or without substituents C_1-8Amino, with or without substituents C_1-8Ester group, with or without substituents C_1-8Aminocarbonyl, with or without substituents C_1-8Hydrocarbyl, with or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8A heterocyclic hydrocarbon group;

b is absent or 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-, C with or without substituents_1-12With or without substituents C_1-12And C with or without substituents_1-12The heterocyclic hydrocarbon group of (a); wherein R is₁₅、R₁₆Each independently selected from H, C with or without substituents_1-8With or without substituents C_1-8And C with or without substituents_1-8The heterocyclic hydrocarbon group of (a);

x is selected from CR₁₉R₂₀、C(＝O)、S(＝O)、SO₂、NR₂₁(ii) a Wherein R is₁₉、R₂₀Each independently selected from H, cyano, hydroxy, amino, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8A heterocyclic hydrocarbon group of (A), C with or without substituents_1-8Hydrocarbyloxy groups of (a); wherein R is₂₁Selected from H, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8The heterocyclic hydrocarbon group of (a);

R₁、R₆、R₁₀each independently selected from H, C with or without substituents_1-8Hydrocarbon group, cycloalkyl group with or without substituent, heterocycloalkyl group with or without substituent, C with or without substituent_1-6An acyl group;

R₂、R₅each independently selected from: hydrogen, OR₃₃、NR₃₄R₃₅Cyano, halogen, C with or without substituents_1～8Hydrocarbon group, cycloalkyl group with or without substituent, heterocycloalkyl group with or without substituent, C with or without substituent_1-6Acyl, amido with or without substituents; wherein R is₃₃、R₃₄、R₃₅Each independently selected from H, C with or without substituents_1～8A cycloalkyl group with or without substituents, a heterocycloalkyl group with or without substituents;

R₃、R₇、R₈、R₉each independently selected from: H. OR (OR)₂₇、NR₂₈R₂₉Cyano, halogen, nitro, C with or without substituents_1-8A hydrocarbon group, a cycloalkyl group with or without a substituent, a heterocycloalkyl group with or without a substituent, X6S (═ O)_kR₃₀、X6C(＝O)R₃₁(ii) a Wherein k is an integer between 0 and 2; wherein R is₂₇，R₂₈，R₂₉，R₃₀，R₃₁Each independently selected from H, C with or without substituents_1～8Hydrocarbyl, cycloalkyl, heterocycloalkyi; wherein X6 is absent or selected from O, S, NR₃₂(ii) a Wherein R is₃₂Is H, C with or without substituents_1-8A cycloalkyl group with or without substituents, and a heterocycloalkyl group with or without substituents;

R₄selected from H, cyano, carboxyl, C with or without substituents_1-8A hydrocarbon group of (a), a hydrocarbyloxycarbonyl group with or without a substituent group;

a is an integer between 0 and 5 (e.g., 1, 2, 3, 4, 5);

b is an integer between 0 and 3 (e.g., 1, 2, 3);

c is an integer between 0 and 30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9);

d is an integer between 0 and 9 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9).

In another preferred embodiment, a is absent; w is X2C (═ O) X3 where X2 is NR₁₈And X3 is absent, or X3 is NR₁₈And X2 is absent; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxy, C with or without substituents_1-4H is an integer between 1 and 6; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently selected from H, hydroxy, C with or without substituents_1-4I is an integer between 1 and 6; c is 0.

In another preferred embodiment, a is absent; w is absent or is O; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxy, C with or without substituents_1-4H is an integer between 0 and 3; b is O; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently selected from H, hydroxy, C with or without substituents_1-4I is between 0 and 3An integer of (d); c is an integer between 1 and 6.

In another preferred embodiment, a is C (═ O) X1, where X1 is absent or selected from (CR)₁₂R₁₃)_eO, and (CR)₁₂R₁₃)_eS, wherein e is an integer between 0 and 2, R₁₂、R₁₃、R₁₄Each independently is hydrogen or C_1-4Alkyl groups of (a); w is O or NR₁₇Wherein R is₁₇Is H, or C with or without substituents_1-4A hydrocarbon group of (a); y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxy, C with or without substituents_1-4H is an integer between 0 and 3; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently selected from H, hydroxy, C with or without substituents_1-4I is an integer between 0 and 3; b is O; c is an integer between 1 and 4.

In another preferred embodiment, A is SO₂X1, wherein X1 is absent or selected from O, and S; w is O; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxy, C with or without substituents_1-4H is an integer between 1 and 6; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently selected from H, hydroxy, C with or without substituents_1-4I is an integer between 0 and 3; c is 0.

In another preferred embodiment, a is absent; w is NR₁₇Wherein R is₁₇Is H, or C with or without substituents_1-4A hydrocarbon group of (a); y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxy, C with or without substituents_1-4H is an integer between 0 and 3; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently selected from H, hydroxy, C with or without substituents_1-4I is between 0 and 4An integer number; b is O; c is an integer between 1 and 6.

In another preferred embodiment, a is absent; deletion of W; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxy, C with or without substituents_1-4H is an integer between 0 and 3; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently selected from H, hydroxy, C with or without substituents_1-4I is an integer between 0 and 3; b is O; c is an integer between 1 and 10 (preferably between 1 and 6).

In another preferred embodiment, a is C (═ O) X1, where X1 is absent or selected from (CR)₁₂R₁₃)_eO, and (CR)₁₂R₁₃)_eS, wherein e is an integer between 0 and 2, R₁₂、R₁₃、R₁₄Each independently is hydrogen or C_1-4Alkyl groups of (a); w is O, NR₁₇-X2C (═ O) X3, or is absent, wherein R is₁₇Is H, or C with or without substituents_1-4A hydrocarbon group of (a); y is CHX4 (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxy, C with or without substituents_1-4H is an integer of 0 to 3, X4 is cyano, nitro, or hydroxy; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently selected from H, hydroxy, C with or without substituents_1-4I is an integer between 0 and 15; b is O; c is an integer between 0 and 10 (preferably between 0 and 6).

In another preferred embodiment, any of the substituents is selected from the group consisting of: halogen, unsubstituted or halogenated C1-C6 alkyl, unsubstituted or halogenated C1-C6 alkoxy, unsubstituted or halogenated C2-C6 alkoxyalkyl, unsubstituted or halogenated C3-C8 cycloalkyl, unsubstituted or halogenated C2-C6 alkylcarbonyl, unsubstituted or halogenated C1-C6 alkylene-hydroxy, unsubstituted or C1-C6 alkyl-substituted amine.

In another preferred embodiment, a is C (═ O); y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently selected from H, hydroxyl and CN, and H is an integer between 1 and 3; preferably, R₂₂Is H, R₂₃Is CN and h is 1.

In another preferred embodiment, the compound has the structure of formula I':

wherein the groups in formula I' are as defined above.

In another preferred embodiment, the compound has the structure of formula I ″:

wherein the groups in formula I' are as defined above.

In another preferred embodiment, in formula I, I' or formula I ",

represents a single bond.

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

In another preferred embodiment, R₁Selected from: H. and C with or without substituents_1-4An alkyl group.

In another preferred embodiment, R₂、R₅Each independently selected from: hydrogen, and C with or without substituents_1-4An alkyl group.

In another preferred embodiment, R₃Selected from: hydrogen, and C with or without substituents_1-4An alkyl group.

In another preferred embodiment, R₆Selected from: hydrogen, C with or without substituents_1-4Alkyl, OR₂₉Wherein R is₂₉Selected from H, C with or without substituents_1-6An alkyl group.

In another preferred embodiment, R₈Selected from: hydrogen, C with or without substituents_1-4Alkyl, and NR₂₈R₂₉Wherein R is₂₈R₂₉Is H, or C with or without substituents_1-4A hydrocarbon group of (1).

In another preferred embodiment, R₈Selected from: hydrogen, halogen, and C with or without substituents_1-4An alkyl group.

In another preferred embodiment, R₉Selected from: hydrogen, halogen, and C with or without substituents_1-4An alkyl group.

In another preferred embodiment, R₁₀Selected from: hydrogen, halogen, cyano, nitro, and C with or without substituents_1-4An alkyl group.

In another preferred embodiment, R₄Selected from H, cyano, C with or without substituents_1-6An alkyl group.

In a second aspect of the present invention, there is provided a pharmaceutical composition comprising a compound of the first aspect, or a pharmaceutically acceptable salt, prodrug thereof, and a pharmaceutically acceptable carrier.

In another preferred embodiment, the effective amount refers to a therapeutically effective amount or an inhibitory effective amount, preferably 0.01 to 99.99%.

In another preferred embodiment, the pharmaceutical composition further comprises one or more additional antineoplastic agents.

In another preferred embodiment, the pharmaceutical composition is for use in degrading JAK 3.

In another preferred embodiment, the pharmaceutical composition is used for treating diseases related to the activity or expression amount of JAK 3.

In a third aspect of the invention, there is provided the use of a compound according to the first aspect of the invention for:

(a) preparing a medicament for treating diseases related to the activity or expression amount of JAK 3;

(b) preparing a JAK3 targeted degradation agent;

(c) non-therapeutically degrading JAK3 in vitro;

(d) non-therapeutically inhibiting tumor cell proliferation in vitro; and/or

(e) Treating diseases associated with the activity or expression level of JAK 3.

In another preferred embodiment, the disease associated with JAK3 activity or expression is a neoplastic or autoimmune disease.

In another preferred embodiment, the disease associated with JAK3 activity or expression is selected from the group consisting of: organ transplant rejection, xenograft rejection, lupus erythematosus, multiple sclerosis, rheumatoid arthritis, psoriasis, cancer, asthma, atopic dermatitis, type I diabetes and diabetic complications, autoimmune thyroid disorders, ulcerative colitis, crohn's disease, alzheimer's disease, leukemia, lymphoma, multiple myeloma, alopecia areata, vitiligo, and the like.

In a fourth aspect of the invention, there is provided a process for the preparation of a compound of formula I as described in the first aspect of the invention, comprising the steps of:

(a) reacting a compound shown in a formula IV with a compound shown in a formula II in an inert solvent to obtain a compound shown in a formula I;

in the above formulae, each group is as defined above, M₁Is a leaving group.

In another preferred example, the method further comprises the steps of:

(b) reacting a compound of formula III with a compound of formula V in an inert solvent to give a compound of formula II, M₁、M₂Is a leaving group.

In a fifth aspect of the invention, there is provided a method of degrading JAK3, comprising the steps of: administering to a subject an effective amount of a compound of formula I as described in the first aspect of the invention or a pharmaceutically acceptable salt thereof, or administering to a subject an inhibitory effective amount of a pharmaceutical composition as described in the fourth aspect of the invention.

In another preferred embodiment, said degradation is non-therapeutic in vitro.

In another preferred embodiment, when an effective amount of a compound of formula I according to the first aspect of the invention or a pharmaceutically acceptable salt thereof is administered to a subject, the inhibitory effective amount is 0.001-500nmol/L, preferably 0.01-200 nmol/L.

In a sixth aspect of the present invention, there is provided a method of treating a disease associated with JAK3 activity or expression, the method comprising: administering to a subject a therapeutically effective amount of a compound of formula I according to the first aspect of the invention, or a pharmaceutical composition according to the fourth aspect of the invention.

In another preferred embodiment, the subject is a mammal; preferably, the mammal is a human.

It is to be understood that within the scope of the present invention, the above-described features of the present invention and those specifically described below (e.g., in the examples) may be combined with each other to form new or preferred embodiments. Not to be reiterated herein, but to the extent of space.

Detailed Description

The inventors of the present invention have made extensive and intensive studies to prepare a compound having a structure represented by formula I, and found that it has JAK3 degradation activity. And the compound can generate degradation effect on JAK3 at an extremely low concentration, and has extremely excellent activity, so that the compound can be used for treating diseases related to the activity or expression level of JAK3, such as tumors. The present invention has been completed based on this finding.

The invention provides a novel compound and application thereof in degrading tyrosine protein kinase JAK 3. The compounds of the present invention are capable of degrading JAK3, and are useful as JAK3 degradants having a dual role in immunomodulation, and for the treatment of diseases in mammals such as rheumatoid arthritis, psoriasis, psoriatic arthritis, ankylosing spondylitis, atopic dermatitis, keratoconjunctivitis sicca, and Crohn's disease.

Term(s) for

In the present invention, the term "C_1-8The hydrocarbyl group means a functional group containing only carbon and hydrogen atoms, wherein the number of carbon atoms is 1 to 8. The hydrocarbyl group can be regarded as a radical remaining after the corresponding hydrocarbon loses one hydrogen atom, and can be an alkyl group, a cycloalkyl group, an alkenyl group, an alkynyl group or the like; the structure can be straight chain, branched chain or cyclic; may be aliphatic or aromatic.

The term "C_1-6The "alkyl group" means a straight or branched alkyl group having 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, tert-butyl, or the like.

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

The term "halo" as used herein, unless otherwise indicated, includes fluoro, chloro, bromo or iodo.

The compounds of the present invention may contain double bonds. When containing such double bonds, the compounds of the present invention exist in cis, trans, or mixtures thereof.

Halogen as used herein includes fluorine, chlorine, bromine and iodine.

Unless otherwise indicated, alkyl groups and the alkyl portion of alkoxy groups referred to herein may be straight chain, branched chain or cyclic.

In the present invention, the term "cycloalkyl group" refers to a functional group containing both carbon and hydrogen atoms. Including cycloalkyl, cycloalkenyl (containing at least one carbon-carbon double bond), and aryl. They may be monocyclic, bicyclic and polycyclic. They may be spiro or fused rings.

In the present invention, the term "heterocyclic hydrocarbon group" means a functional group containing carbon, hydrogen and at least one heteroatom other than carbon and hydrogen. Including heterocycloalkyl, heterocycloalkenyl (containing at least one carbon-carbon double bond), and heteroaryl. One or more of the ring-forming atoms in the ring is a heteroatom. The heteroatoms may be O, N and S atoms, and various combinations thereof. They may be monocyclic, bicyclic and polycyclic. They may be spiro or fused rings.

In the present invention, the term "substituent" includes, but is not limited to, fluorine, chlorine, bromine, cyano, hydroxyl, amino, C_1-6Hydrocarbyloxy, C_1-6Halogenated hydrocarbon group, C_1-6Acyl radical, C_1-6A sulfonyl group.

The term "hydrocarbyloxy" as used herein refers to an O-hydrocarbyl group, wherein "hydrocarbyl" is as defined above.

The term "alkoxycarbonyl" as used herein refers to a C (═ O) O-hydrocarbyl group, where "hydrocarbyl" is as defined above.

The term "amine" as used herein refers to N (H or hydrocarbyl 1) (H or hydrocarbyl 2), wherein "hydrocarbyl" is as defined above.

The term "aminocarbonyl" as used herein refers to C (═ O) -amine groups, where "amine groups" are as defined above.

The term "amido", as used herein, refers to an N (H or hydrocarbyl) -C (═ O) -hydrocarbyl group, where the "hydrocarbyl group" is as defined above.

As used herein, the terms "comprising," "including," or "including" mean that the various ingredients may be used together in a mixture or composition of the invention. Thus, the terms "consisting essentially of and" consisting of are encompassed by the term "comprising.

In the present invention, the term "pharmaceutically acceptable" ingredient refers to a substance that is suitable for use in humans and/or animals without undue adverse side effects (such as toxicity, irritation, and allergic response), i.e., at a reasonable benefit/risk ratio.

In the present invention, the term "effective amount" refers to an amount of a therapeutic agent that treats, alleviates, or prevents a target disease or condition, or an amount that exhibits a detectable therapeutic or prophylactic effect. The precise effective amount for a subject will depend upon the size and health of the subject, the nature and extent of the disorder, and the therapeutic agent and/or combination of therapeutic agents selected for administration. Therefore, it is not useful to specify an exact effective amount in advance. However, for a given condition, the effective amount can be determined by routine experimentation and can be determined by a clinician.

Herein, unless otherwise specified, the term "substituted" means that one or more hydrogen atoms on a group are replaced with a substituent selected from the group consisting of: halogen, unsubstituted or halogenated C_1-6Alkyl, unsubstituted or halogenated C_2-6Acyl, unsubstituted or halogenated C_1-6Alkyl-hydroxy.

Unless otherwise specified, all occurrences of a compound in the present invention are intended to include all possible optical isomers, such as a single chiral compound, or a mixture of various chiral compounds (i.e., a racemate). In all compounds of the present invention, each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R configuration and the S configuration.

As used herein, the term "compounds of the invention" refers to compounds of formula I. The term also includes various crystalline forms, pharmaceutically acceptable salts, hydrates or solvates of the compounds of formula I.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of the present invention with an acid or base that is suitable for use as a pharmaceutical. Pharmaceutically acceptable salts include inorganic and organic salts. One preferred class of salts is that formed by reacting a compound of the present invention with an acid. Suitable acids for forming the salts include, but are not limited to: inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid, etc., organic acids such as 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, phenylmethanesulfonic acid, benzenesulfonic acid, etc.; and acidic amino acids such as aspartic acid and glutamic acid.

Compounds and pharmaceutically acceptable salts thereof

The present invention relates to compounds of formula I or a pharmaceutically acceptable salt thereof;

wherein:

represents a single bond;

represents a single bond or a double bond;

a is absent or selected from C (═ O), C (═ O) X1-, SOX1-, SO₂X1-, C with or without substituents_1-8Hydrocarbyl, C with or without substituents_1-8Cycloalkyl group, and C with or without substituent_1-8A heterocyclic hydrocarbon group; wherein X1 is absent or selected from (CR)₁₂R₁₃)_eO、(CR₁₂R₁₃)_eS and NR₁₄(ii) a Wherein R is₁₂、R₁₃、R₁₄Each independently selected from H, C with or without substituents_1-8With or without substituents C_1-8And C with or without substituents_1-8E is an integer between 0 and 3;

w is absent or selected from O, NR₁₇、-X2C(＝O)X3、-X2S(＝O)_gX3; wherein R is₁₇Is H, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8The heterocyclic hydrocarbon group of (a); wherein X2 and X3 are independently deleted or selected from O, S, NR₁₈(ii) a Wherein g is an integer from 0 to 2; wherein R is₁₈Is H, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8The heterocyclic hydrocarbon group of (a);

y is (CR)₂₂R₂₃)_h、CHX4(CR₂₂R₂₃)_h、CX4＝CH(CR₂₂R₂₃)_hOr (CR)₂₂R₂₃)_h(ii) a Wherein h is an integer between 0 and 30; wherein R is₂₂、R₂₃Each independently selected from H, cyano, hydroxy, amino, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8A heterocyclic hydrocarbon group of (A), C with or without substituents_1-8Hydrocarbyloxy groups of (a); wherein X4 ═ H, halogen, cyano, nitro, hydroxy, with or without substituent C_1-8Hydrocarbyloxy, with or without substituents C_1-8Alkoxycarbonyl, with or without substituents C_1-8Amino, with or without substituents C_1-8Ester group, with or without substituents C_1-8Aminocarbonyl, with or without substituents C_1-8Hydrocarbyl, with or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8A heterocyclic hydrocarbon group;

z is (CR)₂₄R₂₅)_i、CHX5(CR₂₄R₂₅)_i、CX5＝CH(CR₂₄R₂₅)_iOr C ≡ C (CR)₂₄R₂₅)_i(ii) a Wherein i is an integer between 0 and 30; wherein R is₂₄、R₂₅Each independently selected from H, cyano, hydroxy, amino, C with or without substituents_1-8With or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8Heterocycloalkyl radicals, with or without substituents C_1-8A hydrocarbyloxy group; wherein X5 ═ H, halogen, cyano, nitro, hydroxy, with or without substituent C_1-8With or without substituents C_1-8Alkoxycarbonyl, with or without substituents C_1-8Amino, with or without substituents C_1-8Ester group, with or without substituents C_1-8Aminocarbonyl, with or without substituents C_1-8Hydrocarbyl, with or without substituents C_1-8Cycloalkyl, with or without substituents C_1-8A heterocyclic hydrocarbon group;

R₂、R₂each independently selected from: hydrogen, OR₃₃、NR₃₄R₃₅Cyano, halogen, C with or without substituents_1～8Hydrocarbyl, with or without substituentsCycloalkyl, heterocycloalkyl with or without substituent, C with or without substituent_1-6Acyl, amido with or without substituents; wherein R is₅₃、R₃₄、R₃₅Each independently selected from H, C with or without substituents_1～8A cycloalkyl group with or without substituents, a heterocycloalkyl group with or without substituents;

R₃、R₇、R₈、R₆each independently selected from: H. OR (OR)₂₇、NR₂₈R₂₉Cyano, halogen, nitro, C with or without substituents_1-8A hydrocarbon group, a cycloalkyl group with or without a substituent, a heterocycloalkyl group with or without a substituent, X6S (═ O)_kR₃₀、X6C(＝O)R₃₁(ii) a Wherein k is an integer between 0 and 2; wherein R is₂₇，R₂₈，R₂₉，R₃₀，R₃₁Each independently selected from H, C with or without substituents_1～8Hydrocarbyl, cycloalkyl, heterocycloalkyi; wherein X6 is absent or selected from O, S, NR₃₀(ii) a Wherein R is₂₂Is H, C with or without substituents_1-8A cycloalkyl group with or without substituents, and a heterocycloalkyl group with or without substituents;

a is an integer between 0 and 5 (e.g., 1, 2, 3, 4, 5);

b is an integer between 0 and 3 (e.g., 1, 2, 3);

c is an integer between 0 and 30 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9);

d is an integer between 0 and 9 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9).

The integer described herein may be 0, 1, 2, 3, 4, 5, 6, 7, 8, or 9, unless otherwise specified.

In another preferred embodiment, each of the C1-8 cyclic hydrocarbon groups is preferably a C3-8 cyclic hydrocarbon group (e.g., a C3-8 cycloalkyl group).

In another preferred embodiment, each of the C1-8 heterocyclic hydrocarbon groups is preferably a C3-8 heterocyclic hydrocarbon group (e.g., a 4-10 membered heterocycloalkyl group containing 3-8 carbon atoms).

In a preferred embodiment of the invention, the compound is selected from the group consisting of:

the compound package of the present invention may form pharmaceutically acceptable salts with inorganic acids, organic acids or bases. The inorganic acid includes but is not limited to hydrochloric acid, hydrobromic acid, nitric acid, perchloric acid, sulfuric acid or phosphoric acid, etc.; such organic acids include, but are not limited to, methanesulfonic acid, trifluoromethanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, fumaric acid, oxalic acid, acetic acid, maleic acid, ascorbic acid, lactic acid, tartaric acid, malonic acid, glycolic acid, succinic acid, propionic acid, and the like; the base includes, but is not limited to, inorganic salts and amines.

The term pharmaceutically acceptable salts refers to those salts which are, according to medical judgment, suitable for use in contact with the tissues of humans and mammals without excessive toxicity, irritation, allergic response, and the like. Pharmaceutically acceptable salts are well known in the art.

The invention also encompasses pharmaceutical compositions containing prodrugs of compounds of formula I. Prodrugs include compounds wherein the precursor molecule is covalently bonded to a free carboxyl, hydroxyl, amino or amine group of the compound of formula I via a carbonate, carbamate, amide, alkyl ester, phosphate, phosphoramidate linkage.

Preparation of the Compounds

Preparation method

The process for the preparation of the compounds of formula I according to the invention is described in more detail below, but these particular processes do not limit the invention in any way. The compounds of the present invention may also be conveniently prepared by optionally combining various synthetic methods described in the present specification or known in the art, and such combinations may be readily carried out by those skilled in the art to which the present invention pertains.

The following reaction scheme illustrates the preparation of the compounds of the present invention. Unless otherwise indicated, A, B, W, Y, Z, X, a, B, c, d, R in the reaction schemes and discussion that follows₁、R₂、R₃、R₄、R₅、R₆、R₇、R₈、R₉、R₁₀As defined above.

In general, compounds of formula I can be obtained from formula III as described in the following scheme:

in compound II, when W is an ether, it can be prepared by direct nucleophilic substitution of formula III (W1 ═ OH) with an intermediate containing a leaving group under the action of a base, or by reaction with an alcohol using a mitsunobu reaction; when W is ester, carbamate (NHCO)₂) When the compound is prepared, the compound can be prepared by reacting formula III (W1 ═ OH) with acyl chloride, activated ester (amide), carboxylic acid and isocyanate under the action of alkali; when W is an amine, formula III (W1 ═ NH) can be used₂) Direct nucleophilic substitution with an intermediate containing a leaving group under the action of a base can also be prepared by using a compound of formula III (W1 ═ NH)₂) (ii) reductive amination with aldehydes/ketones; when W is an amide, alkoxycarbonylamine (OCONH), or urea, formula III (W1 ═ NH) can be used₂) Reacting with corresponding acyl chloride, activated ester (amide), carboxylic acid and isocyanate under the action of alkali.

In compound I, when A is linked to the nitrogen atom by a C-N bond, it can be prepared by direct substitution or reductive amination; when A is attached to the nitrogen atom by means of an amide, urea, carbamate, sulfonamide, thioamide, it can be prepared from the corresponding acid chlorides, activated esters (amides), carboxylic acids, isocyanates, sulfonyl chlorides, sulfuryl chlorides.

In general, depending on the situation of the linking structures A and W, the compounds I can also be obtained by first linking the formula IV to the intermediate chain and then reacting with the formula III. The chemical synthesis method is the same as that described above.

The compounds of formula III, IV can be obtained by known synthetic methods or readily obtained by commercial purchase.

Use of compounds of formula I

The compounds of formula I may be used for one or more of the following uses:

(b) preparing a JAK3 targeted degradation agent;

(c) non-therapeutically degrading JAK3 in vitro;

(d) non-therapeutically inhibiting tumor cell proliferation in vitro; and/or

(e) Treating diseases related to the activity or expression level of JAK3 (such as autoimmune diseases).

In another preferred embodiment, the disease associated with JAK3 activity or expression is organ transplant rejection, xenotransplantation, lupus erythematosus, multiple sclerosis, rheumatoid arthritis, psoriasis, cancer, asthma, atopic dermatitis, type I diabetes and diabetic complications, autoimmune thyroid disorders, ulcerative colitis, crohn's disease, alzheimer's disease, leukemia, lymphoma, multiple myeloma, alopecia areata, vitiligo, and the like.

The compounds of formula I of the present invention are useful for preparing a pharmaceutical composition comprising: (i) an effective amount of a compound of formula I, or a pharmaceutically acceptable salt thereof; and (ii) a pharmaceutically acceptable carrier.

In another preferred embodiment, the effective amount refers to a therapeutically effective amount or an inhibitory effective amount.

The compounds of formula I of the present invention may also be used in methods of inhibiting or degrading bujak 3, either in vitro non-therapeutic or therapeutic inhibition.

In another preferred embodiment, when an inhibitory effective amount of a compound of formula I of the present invention or a pharmaceutically acceptable salt thereof is administered to a subject, said inhibitory effective amount is 0.001-500nmol/L, preferably 0.01-200 nmol/L.

In particular, the present invention also provides a method of treating a disease associated with JAK3 activity or expression comprising: administering to the subject a therapeutically effective amount of a compound of formula I, or said pharmaceutical composition comprising a compound of formula I as an active ingredient.

Pharmaceutical compositions and methods of administration

The compound can obviously degrade JAK3 so as to play a role in inhibiting JAK3, so that the compound and various crystal forms, pharmaceutically acceptable inorganic or organic salts, hydrates or solvates thereof, and a pharmaceutical composition containing the compound as a main active ingredient can be used for treating, preventing and relieving diseases related to the activity or expression amount of JAK 3. According to the prior art, the compounds of the present invention are useful in the treatment of diseases including tumors and the like.

The pharmaceutical composition of the present invention comprises the compound of the present invention or a pharmacologically acceptable salt thereof in a safe and effective amount range and a pharmacologically acceptable excipient or carrier. Wherein "safe and effective amount" means: the amount of the compound is sufficient to significantly improve the condition without causing serious side effects. Typically, the pharmaceutical composition contains 1-2000mg of a compound of the invention per dose, more preferably, 5-200mg of a compound of the invention per dose. Preferably, said "dose" is a capsule or tablet.

"pharmaceutically acceptable carrier" refers to: one or more compatible solid or liquid fillers or gel substances which are suitable for human use and must be of sufficient purity and sufficiently low toxicity. By "compatible" is meant herein that the components of the composition are capable of intermixing with and with the compounds of the present invention without significantly diminishing the efficacy of the compounds. Examples of pharmaceutically acceptable carrier moieties are cellulose and its derivatives (e.g. sodium carboxymethylcellulose, sodium ethylcellulose, cellulose acetate, etc.), gelatin, talc, solid lubricantsLubricant (such as stearic acid, magnesium stearate), calcium sulfate, vegetable oil (such as soybean oil, sesame oil, peanut oil, olive oil, etc.), polyalcohol (such as propylene glycol, glycerol, mannitol, sorbitol, etc.), emulsifier (such as propylene glycol, glycerol, mannitol, sorbitol, etc.)

) Wetting agents (e.g., sodium lauryl sulfate), coloring agents, flavoring agents, stabilizers, antioxidants, preservatives, pyrogen-free water, and the like.

The mode of administration of the compounds or pharmaceutical compositions of the present invention is not particularly limited, and representative modes of administration include (but are not limited to): oral, intratumoral, rectal, parenteral (intravenous, intramuscular or subcutaneous), and topical administration.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In these solid dosage forms, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, for example, starch, lactose, sucrose, glucose, mannitol and silicic acid; (b) binders, for example, hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose and acacia; (c) humectants, for example, glycerol; (d) disintegrating agents, for example, agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) slow solvents, such as paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, such as cetyl alcohol and glycerol monostearate; (h) adsorbents, for example, kaolin; and (i) lubricants, for example, talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In capsules, tablets and pills, the dosage forms may also comprise buffering agents.

Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings and other materials well known in the art. They may contain opacifying agents and the release of the active compound or compounds in such compositions may be delayed in release in a certain part of the digestive tract. Examples of embedding components which can be used are polymeric substances and wax-like substances. If desired, the active compound may also be in microencapsulated form with one or more of the above-mentioned excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups or tinctures. In addition to the active compounds, the liquid dosage forms may contain inert diluents commonly employed in the art such as, for example, water or other solvents, solubilizing agents and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, propylene glycol, 1, 3-butylene glycol, dimethylformamide and oils, in particular, cottonseed, groundnut, corn germ, olive, castor and sesame oils or mixtures of such materials and the like.

In addition to these inert diluents, the compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents.

Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum methoxide and agar, or mixtures of these substances, and the like.

Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for reconstitution into sterile injectable solutions or dispersions. Suitable aqueous and nonaqueous carriers, diluents, solvents or vehicles include water, ethanol, polyols and suitable mixtures thereof.

Dosage forms for topical administration of the compounds of the present invention include ointments, powders, patches, sprays, and inhalants. The active ingredient is mixed under sterile conditions with a physiologically acceptable carrier and any preservatives, buffers, or propellants which may be required if necessary.

The compounds of the present invention may be administered alone or in combination with other pharmaceutically acceptable compounds.

When the pharmaceutical composition is used, a safe and effective amount of the compound of the present invention is suitable for mammals (such as human beings) to be treated, wherein the administration dose is a pharmaceutically-considered effective administration dose, and for a human body with a weight of 60kg, the daily administration dose is usually 1 to 2000mg, preferably 5 to 500 mg. Of course, the particular dosage will depend upon such factors as the route of administration, the health of the patient, and the like, and is within the skill of the skilled practitioner.

The main advantages of the invention include:

1. provides a compound shown as a formula I.

2. The JAK3 degrading agent with a novel structure and preparation and application thereof are provided, and the inhibitor can degrade JAK3 at an extremely low concentration.

3. Pharmaceutical compositions for treating diseases associated with JAK3 activity are provided.

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out under conventional conditions or conditions recommended by the manufacturers. Unless otherwise indicated, percentages and parts are percentages and parts by weight.

EXAMPLE 1 preparation of Compound 3

The first step is as follows:

2.14g of 6-bromohexanoyl chloride and 1.37g of pomalidomide are dissolved in 50ml of THF (tetrahydrofuran), and the mixture is stirred and refluxed for 8 hours, cooled to room temperature, concentrated and dried in vacuum at 40 ℃ to obtain 2.19g of the compound (1). Ms (esi): 450[ M + H ]]⁺。

The second step is that:

dissolving 1g of compound (1), 1.28g of compound (2) and 2.6g of DIPEA (N, N-diisopropylethylamine) in 50ml of DMF (N, N-dimethylformamide), stirring at 80 ℃ for 6h, cooling to room temperature, concentrating, and performing column chromatography to obtain 810mg of compound (3), yield: 65.6 percent. Ms (esi): 615[ M + H ]]⁺。¹H NMR(400MHz，CD₃OD)δ8.61(d，J＝8Hz，1H)，8.11(s，1H)，7.72(m，1H)，7.54(d，J＝8Hz，1H)，7.10(d，J＝4Hz，1H)，6.63(s，1H)，5.14(dd，J＝12，4Hz，1H)，3.48(br，3H)，3.37(s，1H)，2.50-3.10(m，10H)，2.32(m，1H)，2.16(m，1H)，1.25-2.00(m，9H)，1.03(d，J＝8Hz，3H)。

EXAMPLE 2 preparation of Compound 7

The first step is as follows:

100mg of 4-hydroxythalidomide, 96mg of triethylene glycol monobenzyl ether and 100mg of triphenylphosphine were dissolved in 10ml of anhydrous THF, and 95mg of DIAD (diisopropyl azodicarboxylate) was added dropwise thereto and reacted at room temperature for 2 hours. THF was removed under reduced pressure, and column chromatography was performed to purify the residue to give 110mg of the compound (4). Ms (esi): 497[ M + H]⁺。

The second step is that:

100mg of compound (4) and 100mg of 10% Pd-C were added to 10ml of methanol, and the mixture was hydrogenated at room temperature overnight. Filtration, concentration of the filtrate and purification of the residue by column chromatography gave 40mg of compound (5). Ms (esi): 407[ M + H]⁺。

The third step:

30mg of the compound (5) was dissolved in 5ml of methylene chloride, and 47mg of Dess-martin oxidant was added thereto to conduct a reaction at room temperature for 3 hours. Adding NaHCO₃Saturated aqueous solution and Na₂S₂O₃Stirring for 5min to obtain organic layer, and treating with anhydrous Na₂S₂O₃Drying and concentrating to dryness to obtain the compound (6) which is directly used for the next reaction.

After dissolving Compound (6) in 7ml of methylene chloride, 36mg of the starting Material (2) and NaBH (OAc) were added₃23mg, at room temperature overnight, and after dichloromethane was distilled off under reduced pressure and purified by column chromatography, compound (7) was obtained, 20mg. ms (ESI): 634[ M + H ]]⁺。¹H NMR (hydrochloride, 400MHz, D)₂O)δ8.09(d，J＝1.6Hz，1H)，7.45-7.62(m，1H)，7.18(dd，J＝12.0，7.2Hz，1H)，7.07(m，2H)，6.54-6.70(m，1H)，4.90-5.08(m，2H)，4.12(m，2H)，3.30-3.85(m，14H)，3.20(s，3H)，2.73(m，2H)，2.42(m，2H)，2.09(m，2H)，1.73(m，1H)，1.00(d，J＝8.0Hz，3H)。

EXAMPLE 3 preparation of Compound 10

The first step is as follows:

benzyl 2- (2- (2-bromoethoxy) ethoxy) acetate 174mg, potassium carbonate 100mg, potassium iodide 20mg and pomalidomide 100mg were added to 20ml of DMF (N, N-dimethylformamide) and reacted at 80 ℃ overnight. The reaction solution was purified by column chromatography to give 113mg of the compound (8). Ms (esi): 510[ M + H]⁺。

The second step is that:

100mg of Compound (8) and 10% Pd-C100mg were added to 10ml of methanol, and the mixture was hydrogenated at room temperature overnight. Filtration, concentration of the filtrate and purification of the residue by column chromatography gave 73mg of compound (9). MS (ESI negative ion): 418[ M-H ]]^-。

The third step:

50mg of the compound (9) and 30mg of the starting material (2) were dissolved in 5ml of methylene chloride, and 20mg of HOBt (1-hydroxybenzotriazole) and 40mg of EDC (1- (3-dimethylaminopropyl) -3-ethylcarbodiimide hydrochloride) were added to the solution to conduct a reaction at room temperature overnight. Dichloromethane was distilled off under reduced pressure, and column chromatography purification was performed to obtain compound (10), 50mg. ms (ESI): 647[ M + H ]]⁺。¹H NMR(400MHz，CDCl3)δ10.63(br，1H)，9.84(br，1H)，8.22(d，J＝1.2Hz，1H)，7.43(t，J＝7.6Hz，1H)，7.06(dd，J＝7.2，1.2Hz，1H)，6.97(d，J＝2.8Hz，1H)，6.82(d，J＝8.4Hz，1H)，6.51(d，J＝3.6Hz，1H)，6.42(m，1H)，5.12(br，1H)，4.91(m，1H)，4.55(br，2H)，3.89(m，4H)，3.64(s，3H)，3.51(br，2H)，3.38(m，3H)，2.50-3.01(m，6H)，2.25(br，1H)，2.10(m，1H)，1.86(br，1H)，1.64(br，1H)，0.96(d，J＝6.8Hz，3H)。

EXAMPLE 4 preparation of Compound 13

The first step is as follows:

200mg of the starting material (2) was dissolved in 20ml of methylene chloride, and 100mg of triethylamine and 180mg of 5-benzyloxypentanesulfonyl chloride were added to the solution to conduct a reaction at room temperature overnight. The solvent was distilled off under reduced pressure, and column chromatography was performed to purify the residue to obtain 187mg of compound (11). Ms (esi): 486[ M + H]⁺。

The second step is that:

180mg of compound (11) and 180mg of 10% Pd-C were added to 20ml of methanol, and the mixture was hydrogenated at room temperature overnight. Filtering, concentrating the filtrate, and purifying by column chromatography to obtain compound (12)134mg. MS (ESI): 396[ M + H]⁺。

The third step:

50mg of 4-hydroxythalidomide, 50mg of triphenylphosphine and 50mg of compound (12) were dissolved in 5ml of anhydrous tetrahydrofuran, and 50mg of DIAD (diisopropyl azodicarboxylate) was added dropwise and reacted at room temperature for 3 hours. The tetrahydrofuran was removed under reduced pressure, and column chromatography was performed to purify the residue to obtain 33mg of the compound (13). Ms (esi): 652[ M + H]⁺。¹H NMR (hydrochloride, 400MHz, D)₂O)δ8.11(d，J＝2.0Hz，1H)，7.43-7.61(m，1H)，7.20(dd，J＝11.6，6.8Hz，1H)，7.08(m，2H)，6.60-6.71(m，1H)，5.02(m，2H)，4.54(m，1H)，4.02(m，2H)，3.85(m，2H)，3.18(s，3H)，2.72-3.12(m，4H)，2.15-2.50(m，4H)，1.28-2.06(m，9H)，0.96(d，J＝7.6Hz，3H)。

EXAMPLE 5 preparation of Compound 17

First step of

4-Fluorothalidomide (2.00g) was added to 3-amino-1-propanol (10.00g), microwave-reacted for 0.5 hour, the reaction solution was diluted with dichloromethane (50ml), washed successively with 0.5M dilute hydrochloric acid, water, saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified with a silica gel column to give 0.60g of Compound 14 as a yellow solid in yield: 25 percent. Ms (esi): 331[ M + H]⁺。

Second step of

Compound 17(200mg) and diisopropylethylamine (150mg) were dissolved in diisopropylethylamine (Dietz. RTM. TM.) solutionTo methyl chloride (10ml), methanesulfonyl chloride (80mg) was added dropwise, and after the addition, the reaction solution was stirred overnight. The reaction mixture was washed with 0.5M dilute hydrochloric acid and saturated brine, the organic phase was concentrated under reduced pressure, the residue was dissolved in anhydrous N, N-dimethylformamide (10ml), tofacitinib free base 16(226mg) was added thereto, and 60% sodium hydride (40mg) was added in portions under cooling in an ice-water bath, slowly warmed to room temperature, and stirred overnight. The reaction was concentrated under reduced pressure and the residue was purified on silica gel preparation plates to give 16mg of compound 17 as a yellow solid in yield: 4.2 percent. Ms (esi): 626[ M + H]⁺。

Similarly, the following compounds were prepared by methods analogous to the above examples:

example 6 Western blot to detect JAK3 protein degrading Activity of Compounds

Cell lines: jurkat cell line was cultured in RPMI1640 containing 10% calf serum at 37 ℃ in 5% CO₂And culturing in an incubator with saturated humidity.

A DMSO control group and a compound dry group (10 mu M) are arranged, cells are collected after 24 hours of treatment, 100 mu L of precooled cell lysate is added, the cells are lysed for 30min on ice, total cell protein is extracted, and the protein concentration is determined and quantified by a bicinchoninic acid (BCA) method. Preparing gel conventionally, loading, performing electrophoresis, then transferring a membrane, sealing, respectively adding rabbit anti-human JAK3 (1: 500), incubating overnight at 4 ℃, adding horseradish peroxidase labeled goat anti-rabbit IgG (1: 5000) after rinsing, developing with ECL developer after rinsing, scanning and imaging by a Bio-Rad gel imaging system, and processing and analyzing by computer software. The control was referenced to glyceraldehyde phosphate dehydrogenase (GAPDH).

Grey scale analysis was performed on each band using Image J software to calculate the degradation rate of the compound to degrade JAK3 protein.

The degradation activity of the compounds on JAK3 protein in Jurkat cells is shown in the following table:

compound numbering	Degradation activity	Compound numbering	Degradation activity
				3	++++	22	++++
7	+++	23	++++
				10	+++	24	+++
14	+++	25	+++
				15	++++	26	++++
18	+++	27	++++
				19	+++	28	+++
20	+++	29	+++
				21	+++	30	++++

Remarking: in the above tables, "-" represents no degradation activity, "+" represents a degradation rate of 10% -30%, "+ +" represents a degradation rate of 30% -50%, "+ + + + + + +" represents a degradation rate of 50% -90%, and "+ + + + + + +" represents a degradation rate of more than 90%.

Example 7 CCK8 assay for inhibition of Jurkat cell proliferation (human leukemia T lymphocytes)

The inhibitory effect of the compounds on the proliferation of Jurkat cells was tested in vitro using the CCK8 assay. The method comprises the following specific steps:

culturing Jurkat cells in 1640 culture medium containing 10% calf serum, inoculating to 96-well plate, and culturing at 2 × 10⁵Cells/well, 5% CO at 37 ℃₂In an incubator. Dissolving the compound in dimethyl sulfoxide (DM)SO), 10mM solution was obtained, diluted with phosphate buffer to the desired concentration, and added to the above 96-well plate at 2 wells each, 10. mu.l per well, and each concentration was subjected to two parallel tests. DMSO was diluted accordingly in a gradient and added to the plate as a control.

Subjecting the 96-well plate to 5% CO at 37 deg.C₂After culturing for 48 hours in the cell culture box, 10 μ l of CCK8 solution is added to each well, and the incubation is continued for 1-4 hours in the culture box. The 465nm light absorption value was measured.

The relative viability of the cells after compound treatment was calculated from the light absorption values.

IC of compound on Jurkat cells was calculated by software₅₀。

The in vitro proliferation inhibitory effect of the synthesized compounds on Jurkat cells is shown in table 1:

TABLE 1

Compound numbering	IC₅₀(μM)
		3	20
7	100
		10	60
14	10
		15	8

The results show that the synthesized compound has good activity of inhibiting Jurkat cells.

EXAMPLE 8 in vivo anti-rheumatoid arthritis Activity of Compound (15)

The anti-rheumatoid arthritis effect of the compound (15) was determined in vivo using a Wistar rat rheumatoid arthritis induced by Freund's adjuvant as a model. The method comprises the following specific steps:

1) test sample

Sample preparation: compound (15).

2) Preparation method

Sample preparation: compound (15) dissolved in a physiological saline injection solution at the time of preparation

3) Animals and adjuvants

Wistar rats 24, male, weighing 170-200 g.

Adjuvant: complete Freund's Adjuvant (CFA), Sigma.

4) Test method

Wistar rats were randomly divided into 4 groups of 6 rats each. Blank control group, model group, compound (15) group (20mg/kg ip administration), and tofacitinib positive control group (20mg/kg po administration), respectively. Rheumatoid arthritis was induced by intradermal injection of CFA 0.15mL into the plantar left hind paw of the rat. The rats started dosing treatment on day 11 after model building, once a day, and the treatment is finished on day 20. After the treatment is finished, the volume of the left hind paw is measured, and the paw edema inhibition rate is calculated.

5) Test results

The inhibition rate of compound (15) on the foot swelling of rats induced by Freund's complete adjuvant is shown in Table 2:

TABLE 2

Group of	Foot swelling inhibition ratio (%)
		Blank control group	-
Model set	-
		Compound (15)	49
Positive control (tofacitinib)	29

The in vivo experiment results show that the compound (15) has good anti-rheumatoid arthritis activity in vivo, and the activity is obviously higher than that of a positive control medicament tofacitinib.

All documents referred to herein are incorporated by reference into this application as if each were individually incorporated by reference. Furthermore, it should be understood that various changes and modifications of the present invention can be made by those skilled in the art after reading the above teachings of the present invention, and these equivalents also fall within the scope of the present invention as defined by the appended claims.

Claims

1. A compound of formula I, or a pharmaceutically acceptable salt thereof:

wherein:

represents a single bond;

represents a single bond;

a is absent or selected from C (═ O) X1; wherein X1 is (CR)₁₂R₁₃)_eO; wherein R is₁₂、R₁₃Each independently is H, e is an integer between 0 and 3;

w is absent or selected from O, NR₁₇X2C (═ O) X3; wherein R is₁₇Is H; wherein X3 is NR₁₈And X2 is absent, R₁₈Is H;

y is (CR)₂₂R₂₃)_h(ii) a Wherein h is an integer between 0 and 6; wherein R is₂₂、R₂₃Each independently is H;

z is (CR)₂₄R₂₅)_i(ii) a Wherein i is an integer between 0 and 6; wherein R is₂₄、R₂₅Each independently is H;

b is absent or is O;

x is C (═ O);

R₁is hydrogen; r₆Is methyl; r₁₀Is hydrogen; r₂Is hydrogen; r₅Is methyl;

R₃、R₇、R₈、R₉each independently is hydrogen; r₄Is hydrogen;

a is 5; b is 3; c is an integer between 0 and 6; d is 1.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein:

in said formula I, A is absent; w is X2C (═ O) X3 where X3 is NR₁₈And X2 is absent; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, H is an integer between 1 and 6; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is an integer between 1 and 6; c is 0; or

In said formula I, A is absent; w is absent or is O; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, and H is an integer from 0 to 3; b is O; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is an integer between 0 and 3; c is an integer between 1 and 6; or

Wherein a is C (═ O) X1, where X1 is (CR)₁₂R₁₃)_eO; wherein R is₁₂、R₁₃Each independently is H, e is 1; w is NR₁₇Wherein R is₁₇Is H; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, and H is an integer from 0 to 3; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is an integer between 0 and 3; b is O; c is an integer between 1 and 4; or

In said formula I, A is absent; w is NR₁₇Wherein R is₁₇Is H; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, and H is an integer from 0 to 3; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is an integer between 0 and 4; b is O; c is an integer between 1 and 6; or

In said formula I, A is absent; deletion of W; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, H is 0; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is 0; b is O; c is an integer between 1 and 6.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein in formula I, a is absent; w is X2C (═ O) X3, where X3 is NH and X2 is absent; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, H is an integer between 1 and 6; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is 0(ii) a c is 0.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a is absent; deletion of W; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, and H is an integer from 0 to 3; b is O; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is an integer between 0 and 3; c is an integer between 1 and 6.

5. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein a is absent; deletion of W; y is (CR)₂₂R₂₃)_hWherein R is₂₂、R₂₃Each independently is H, H is 0; z is (CR)₂₄R₂₅)_iWherein R is₂₄、R₂₅Each independently is H, i is 0; b is O; c is an integer between 1 and 4.

6. The compound of claim 1, or a pharmaceutically acceptable salt thereof, selected from the group consisting of:

7. use of a compound of claim 1, or a pharmaceutically acceptable salt thereof, for:

(b) preparing a JAK3 targeted degradation agent;

(c) non-therapeutically degrading JAK3 in vitro; and/or

(d) Non-therapeutically inhibiting tumor cell proliferation in vitro.

8. A method of degrading JAK3 in vitro, the method comprising: administering to a subject an effective amount of a compound of formula I as described in claim 1 or a pharmaceutically acceptable salt thereof.