Tricyclic compounds as BTK kinase inhibitors and uses thereof

Abstract

The invention relates to a BTK kinase inhibitor shown as a formula I, definitions of substituent groups in # imgabs0 # are as described in the specification, and the BTK kinase inhibitor is used for treating autoimmune diseases, inflammation or tumor-related diseases.

Description

Technical Field

[0001] The present invention relates to a tricyclic compound having an inhibitory effect on BTK kinase and a use thereof. Background Art

[0002] BTK belongs to the non-receptor tyrosine kinase TEC (tyrosine kinase expressed in hepatocellular carcinoma) kinase family. The human BTK gene is 37.5kb in length on the chromosome, encoding 659 amino acids. The BTK kinase structure contains 5 main domains, starting from the N-terminus, namely the PH domain, TH domain, SH3 domain, SH2 domain and Kinase domain.

[0003] The PH domain, TH domain, SH3 domain, SH2 domain and Kinase domain recognize and bind to a variety of signaling molecules, allowing BTK protein to participate in a variety of intracellular signaling pathways. The PH domain recognizes and binds to PIP3 to recruit BTK to the cell membrane and participate in signal transduction triggered by extracellular stimuli; the proline-rich region of the TH domain can cause internal folding of the molecule; the SH3 domain can interact with molecules such as Syk, and the Y233 site on the domain can undergo autophosphorylation; the SH2 domain can phosphorylate tyrosine, affecting the signal transduction of downstream cells; the Kinase domain, also known as the catalytic domain, is highly conserved in evolution. The phosphorylation of the highly conserved tyrosine residue (Y551 of BTK) by the Src family kinase and the autophosphorylation of Y233 occur simultaneously, and the BTK kinase will be fully activated.

[0004] BTK kinase is a key kinase in the BCR signaling pathway. After being activated, BTK kinase transduces signals to downstream signaling molecules and phosphorylates PLCγ2, thereby affecting the intracellular calcium ion level and activating the NFκB signaling pathway. It can also be activated by PKCβ and affect cell proliferation. Therefore, the BCR signaling pathway plays an important role in the process of cell growth and proliferation by regulating various kinases.

[0005] BTK kinase is highly expressed in different types of malignant blood tumors. In addition to being highly expressed in blood tumors, BTK kinase is also involved in solid tumors. BTK is also expressed in some tumors such as prostate cancer and colon cancer. In addition to anti-tumor effects, it is also closely related to autoimmune diseases such as arthritis. BTK is involved in the development and maturation of B cells. BTK kinase deficiency in the human body can cause serious immunodeficiency diseases such as X-linked agammaglobulinemia. Because BTK is involved in the proliferation and apoptosis of B cells, when the body has an immune response, B cell signals are activated by activating BTK, thereby regulating the growth and proliferation of B cells; in cells, BTK kinase can regulate transcription factors such as BAP-135 and transcriptional activity, thereby regulating the expression of proteins related to B cell proliferation and apoptosis; BTK kinase can also regulate the expression of immunoglobulins by activating ARID family transcription factors. BTK is also involved in allergic reactions. When cells are first stimulated by antigens, B cells produce IgE, and the Fc segment of IgE can bind to the high-affinity receptor FCεRI. When the cells are stimulated by the same antigen again, they combine with the IgE produced for the first time, activating the FCεRI on the cell surface. The Syk kinase in the BCR signaling pathway is activated, thereby activating the BCR signaling pathway. Ultimately, the cells release proinflammatory mediators, mediating the occurrence of allergic reactions.

[0006] BTK is also related to inflammatory response. BTK is overexpressed in human mononuclear B cells, which promotes the production of TNF-α factor. When the BTK gene is abnormal, the TNF-α factor in the body is reduced, causing an inflammatory response. In addition, the activation of BTK kinase can also induce the production of pro-inflammatory factors in macrophages, thereby causing the occurrence of autoimmune diseases.

[0007] In summary, BTK kinase is a very promising target for the treatment of tumors, autoimmune diseases and inflammatory responses. Although drugs targeting this target have been developed and marketed, current therapeutic drugs still have problems such as drug resistance and low response rate, and new compounds are needed to provide new treatment options for patients. Summary of the invention

[0008] The present invention provides a BTK kinase inhibitor. Compounds of this structure have the effects of good activity, improved drug resistance, less toxic and side effects, good liver microparticle stability, good permeability and high brain penetration rate.

[0009] The present invention relates to a compound of formula I and its stereoisomers, solvates, pharmaceutically acceptable salts or prodrugs:

[0010]

[0011] Wherein, A is selected from:

[0012]

[0013] R1a is selected from H, deuterium, halogen, amino, nitro, mercapto, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy;

[0014] R1 is selected from H, deuterium, halogen, amino, nitro, sulfhydryl, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-8 cycloalkyl, C3-8 heterocyclyl, C5-10 aryl, C5-10 heteroaryl, wherein the C3-8 cycloalkyl, C3-8 heterocyclyl, C5-10 aryl, C5-10 heteroaryl are optionally substituted with deuterium, halogen, amino, nitro, sulfhydryl, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy;

[0015] R2 is selected from H, deuterium, halogen, amino, nitro, mercapto, cyano, carboxyl, C1-6 alcohol, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-8 cycloalkyl, C3-8 heterocyclyl, C5-10 aryl, C5-10 heteroaryl; R3 is selected from O, N, S or (CH2)n, wherein n is an integer from 0 to 6;

[0016] R4 is selected from a 5-10 membered monocyclic heterocyclic group or heteroaryl group, wherein the monocyclic heterocyclic group or heteroaryl group contains 2-4 heteroatoms selected from N, O, and S; optionally, the 5-10 membered heterocyclic group or heteroaryl group is substituted by H, halogen, amino, nitro, thiol, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, or C1-6 hydroxyalkoxy.

[0017] In some embodiments, the R1a is H, C1-6 alkyl; more preferably H, methyl.

[0018] In some embodiments, R1 is H, deuterium, halogen, amino, nitro, thiol, cyano, carboxyl, C1-6 alkyl; more preferably, it is halogen, and the halogen is selected from F, Cl, Br, I; most preferably, it is F.

[0019] In some embodiments, R2 is H, C1-6 alcohol; preferably H, tert-butyl alcohol.

[0020] In some embodiments, R3 is O, NH, (CH2)n, wherein n is 0.

[0021] In some embodiments, R4 is

[0022] In another aspect, the present invention provides a compound as shown in Formula II and a stereoisomer, solvate, pharmaceutically acceptable salt or prodrug thereof:

[0023]

[0024] wherein R1a is methyl or hydrogen, R1b is F or hydrogen, R1c is F or hydrogen, R2 is tert-butyl alcohol or hydrogen, R3 is -NH- or -O-, and R4 is

[0025] In another aspect, the present invention provides a compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug having the structure as described below:

[0026]

[0027]

[0028] In another aspect, the present invention provides a pharmaceutical composition comprising a therapeutically effective dose of the above-mentioned compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug and one or more pharmaceutically acceptable carriers or excipients.

[0029] In another aspect, the present invention provides use of the above-mentioned compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug, or the above-mentioned pharmaceutical composition in the preparation of a Bruton's tyrosinase inhibitor.

[0030] In another aspect, the present invention provides use of the above-mentioned compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug, or the above-mentioned pharmaceutical composition in the preparation of a drug for autoimmune disease, inflammation or tumor.

[0031] Autoimmune diseases include, but are not limited to, inflammatory bowel disease (e.g., ulcerative colitis), arthritis, lupus, rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Ord's thyroiditis, Graves' disease, Sjogren's syndrome, s syndrome, Sjogren's dry eye, non-Sjogren's dry eye, multiple sclerosis, Guillain-Barre syndrome, acute disseminated encephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome, ankylosing spondylitis, antiphospholipid antibody syndrome, aplastic anemia, autoimmune hepatitis, coeliac disease, Goodpasture's syndrome syndrome, idiopathic thrombocytopenic purpura, optic neuritis, scleroderma, primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis, warm autoimmune hemolytic anemia, Wegener's granulomatosis, psoriasis, alopecia universalis, Behcet's disease, chronic fatigue, familial autonomic insufficiency, endometriosis, interstitial cystitis, neuromyotonia, scleroderma, and vulvodynia.

[0032] Inflammatory diseases include, but are not limited to, asthma, appendicitis, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, colitis, conjunctivitis, cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epididymitis, fasciitis, fibrositis, gastritis, gastroenteritis, hepatitis, hidradenitis suppurativa, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, pancreatitis, mumps, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, pneumonia, proctitis, prostatitis, pyelonephritis, rhinitis, eustachian tube inflammation, sinusitis, stomatitis, synovitis, tendonitis, tonsillitis, uveitis, vaginitis, vasculitis, or vulvitis. Or the patient in need or recognized need thereof is afflicted with an inflammatory skin disease such as dermatitis, contact dermatitis, eczema, urticaria, rosacea, and cicatricial psoriatic lesions in the skin, joints, or other tissues or organs.

[0033] Tumors include, but are not limited to, diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, B-ALL, B-cell prolymphocytic leukemia, small lymphocytic lymphoma (SLL), multiple myeloma, B-cell non-Hodgkin lymphoma, lymphoplasmacytic lymphoma / Waldenstrom macroglobulinemia, splenic marginal zone lymphoma, plasma cell myeloma, plasmacytoma, extranodal marginal zone B-cell lymphoma, nodal marginal zone B-cell lymphoma, mantle cell lymphoma, mediastinal (thymic) large B-cell lymphoma, intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma / leukemia, and lymphomatoid granuloma. For example, brain tumors include gliomas, brain stem gliomas, astrocytomas of the cerebellum or cerebrum, malignant gliomas, ependymomas, oligodendrogliomas, meningiomas, craniopharyngiomas, hemangioblastomas, medulloblastomas, visual pathway and hypothalamic gliomas or malignant gliomas; the astrocytoma of the cerebellum or cerebrum is a fibrillary cell astrocytoma or a diffuse astrocytoma or an anaplastic (malignant) astrocytoma.

[0034] The above-mentioned compound or pharmaceutical composition can be administered by nasal or oral inhalation, oral administration, subcutaneous administration, intravenous administration or intramuscular administration.

[0035] In another aspect, the present invention provides a method for preparing a compound of formula III, comprising step (1):

[0036] Alternatively, it further comprises step (2):

[0037]

[0038] Or the preparation method of the compound of formula III comprises the steps:

[0039] Or further comprising the steps:

[0040]

[0041] Both of the above methods utilize the Suzuki coupling reaction. Under the catalysis of a palladium complex and the action of a base, boric acid or boric ester (III-12, III-13) undergoes a coupling reaction with a halogenated aromatic hydrocarbon (III-11) to obtain the final product (III) or form an intermediate (III-14), which is further reacted to obtain the final product.

[0042] Or the preparation method of the compound of formula III also includes the steps of:

[0043]

[0044] The preparation method is through a Buchwald-Hartwig coupling reaction, in which an aryl halide (III-15) and an amine (A-R3-R4) undergo a coupling reaction under the action of a palladium catalyst and a base to form an intermediate (III-16), and the intermediate further hydrolyzes the cyano group to form a final product.

[0045] Terminology

[0046] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0047] In the present invention, alkyl refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2, 3-Dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethylpentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers thereof.

[0048] More preferred are lower alkyl groups having 1 to 6 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like.

[0049] The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available attachment point. The substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate groups. Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl and hydroxy-substituted alkyl are preferred in the present invention; the hydroxy-substituted alkyl may be 2-hydroxyisopropyl or 1-hydroxyethyl.

[0050] In the present invention, non-limiting examples of the C1-6 alcohol include methanol, ethanol, propanol, isopropanol, n-butanol, isobutanol, sec-butanol, tert-butanol, 1-pentanol, 2-pentanol, 3-pentanol, 1-hexanol, 2-hexanol or any combination thereof.

[0051] In the present invention, the cycloalkyl group refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, and the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 8 carbon atoms, and further preferably 3 to 6 carbon atoms.

[0052] Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, and the like; polycyclic cycloalkyls include spirocycloalkyls, fused cycloalkyls, and bridged cycloalkyls.

[0053] The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, etc. The cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0054] In the present invention, a heterocyclic group refers to a saturated or partially unsaturated monocyclic or polycyclic heterocyclic group containing 3 to 20 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, but does not include -OO-, -OS- or -SS- ring parts, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 10 ring atoms; and further preferably, it contains 3 to 8 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, pyrrolidonyl, piperidin-2-onyl, 3,4-dihydropyridine-2 (1H)-onyl, 4,5-dihydropyridazine-3 (2H)-onyl, azetidinyl, oxetanyl, oxanyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc.; preferably pyrrolidyl, pyrrolidinone, piperidin-2-one, 3,4-dihydropyridine-2 (1H) -one, 4,5-dihydropyridazine-3 (2H) -one, azetidinyl, oxetanyl, dihydropyrrolyl, tetrahydrofuranyl, pyrazolidinyl, morpholinyl,

[0055] more preferably, dihydropyrrolyl, pyrrolidyl, pyrrolidonyl, piperidin-2-onyl, 3,4-dihydropyridin-2(1H)-onyl, 4,5-dihydropyridazin-3(2H)-onyl, azetidinyl, oxetanyl, oxanyl, morpholinyl, piperidinyl, piperazinyl, Pyranyl. Polycyclic heterocyclic groups include spirocyclic, condensed and bridged heterocyclic groups; wherein the spirocyclic, condensed and bridged heterocyclic groups are optionally connected to other groups through single bonds, or further connected to other cycloalkyl, heterocyclic, aryl and heteroaryl groups through any two or more atoms on the ring.

[0056] The heterocyclyl ring may be fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring attached to the parent structure is a heterocyclyl, non-limiting examples of which include:

[0057]

[0058] The heterocyclyl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

[0059] In the present invention, aryl refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings sharing adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, more preferably 6- to 8-membered, such as phenyl and naphthyl, preferably phenyl.

[0060] The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0061] In the present invention, heteroaryl refers to a heteroaromatic system comprising 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 yuan, more preferably 5 to 8 yuan, and most preferably 5 or 6 yuan, such as pyrazinyl, pyridazinyl, imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, isoxazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, oxadiazole, pyrazinyl, etc., preferably pyrimidinyl, pyrazolyl, oxazolyl, isoxazolyl, oxadiazole, pyridine.

[0062] The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0063] In the present invention, alkoxy refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as described above, preferably an alkyl containing 1 to 8 carbon atoms, more preferably an alkyl containing 1 to 6 carbon atoms, and most preferably an alkyl containing 1 to 3 carbon atoms. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate;

[0064] Non-limiting examples of alkoxy also include propan-2-oxy and the like.

[0065] In the present invention, haloalkyl refers to an alkyl group substituted by one or more halogens, wherein alkyl is as defined above. Non-limiting examples of haloalkyl include: trifluoromethyl, trifluoroethyl;

[0066] Non-limiting examples of haloalkyl also include difluoromethyl, 1,1,2,2-tetrafluoroethyl, perfluoroethyl, and the like.

[0067] In the present invention, haloalkoxy refers to an alkoxy group substituted by one or more halogens, wherein alkoxy is as defined above;

[0068] The halogenated alkoxy group may be fully halogenated or partially halogenated, and the number of halogenations may be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, etc.; the halogen is preferably F, Cl, Br, I; for example, it may be trifluoromethoxy, difluoromethoxy, 1,1,2,2-tetrafluoroethoxy, perfluoroethoxy, etc.

[0069] In the present invention, hydroxyalkyl refers to an alkyl group substituted by a hydroxy group, wherein the alkyl group is as defined above.

[0070] In the present invention, alkenyl refers to alkenyl, also known as olefin group, preferably alkenyl containing 2 to 8 carbon atoms, more preferably alkenyl containing 2 to 6 carbon atoms, further preferably alkenyl containing 2 to 4 carbon atoms, and most preferably alkenyl containing 2 to 3 carbon atoms. Non-limiting examples of alkenyl include: vinyl, propenyl. The alkenyl group described herein can be further substituted by other related groups, for example: alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0071] In the present invention, alkynyl refers to chain alkynyl, also known as alkyne, which refers to an unsaturated hydrocarbon group containing -C≡C-; preferably an alkynyl containing 2 to 8 carbon atoms, more preferably an alkynyl containing 2 to 6 carbon atoms, further preferably an alkynyl containing 2 to 4 carbon atoms, and most preferably an alkynyl containing 2 to 3 carbon atoms. The alkynyl group may be further substituted by other related groups, such as alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

[0072] In the present invention, haloalkyl refers to an alkyl group substituted by one or more halogens, wherein alkyl is as defined above.

[0073] In the present invention, haloalkoxy refers to an alkoxy group substituted by one or more halogens, wherein the alkoxy group is as defined above.

[0074] In the present invention, hydroxyalkyl refers to an alkyl group substituted by a hydroxy group, wherein the alkyl group is as defined above.

[0075] In the present invention, a hydroxyl group refers to an -OH group.

[0076] In the present invention, halogen means fluorine, chlorine, bromine or iodine.

[0077] In the present invention, amino group refers to -NH2.

[0078] In the present invention, cyano refers to -CN.

[0079] In the present invention, nitro refers to -NO2.

[0080] In the present invention, the carboxyl group refers to -C(O)OH.

[0081] The hydrogen atoms described in the present invention can be replaced by their isotope deuterium, and any hydrogen atom in the example compounds of the present invention can also be replaced by a deuterium atom.

[0082] In the present invention, "optional" or "optionally" means that the event or environment described later can but need not occur. For example, "optionally a heterocyclic group substituted with an alkyl group" means that an alkyl group can but need not exist, and the description includes the situation where the heterocyclic group is substituted with an alkyl group and the situation where the heterocyclic group is not substituted with an alkyl group.

[0083] In the present invention, "substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogen may be unstable when combined with carbon atoms with unsaturated (such as olefinic) bonds.

[0084] The compounds of this patent application include isotopic derivatives thereof. The term "isotopic derivative" refers to a compound whose structure differs only in the presence of one or more isotopically enriched atoms. For example, compounds having the structure of this patent application, replacing hydrogen with "deuterium" or "tritium", or replacing fluorine with 18F-fluorine labeling (18F isotope), or replacing carbon atoms with 11C-, 13C-, or 14C-enriched carbon (11C-, 13C-, or 14C-carbon labeling; 11C-, 13C-, or 14C-isotope) are within the scope of this patent application. Such compounds can be used as analytical tools or probes in, for example, biological assays, or can be used as in vivo diagnostic imaging tracers for diseases, or as tracers for pharmacodynamics, pharmacokinetics, or receptor studies. The various deuterated forms of compounds of this patent application refer to compounds in which each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize deuterated forms of compounds with reference to relevant literature. Commercially available deuterated starting materials can be used to prepare deuterated forms of the compounds, or they can be synthesized using conventional techniques using deuterated reagents, including but not limited to deuterated borane, trideuterated borane tetrahydrofuran solution, deuterated lithium aluminum hydride, deuterated iodoethane and deuterated iodomethane, etc. Deuterated compounds can generally retain activities comparable to those of undeuterated compounds, and when deuterated at certain specific sites, better metabolic stability can be achieved, thereby obtaining certain therapeutic advantages.

[0085] The compounds of this patent application may exist in specific stereoisomer forms. The term "stereoisomer" refers to isomers with the same structure but different arrangements of atoms in space. It includes cis and trans (or Z and E) isomers, (-)- and (+)-isomers, (R)- and (S)-enantiomers, diastereomers, (D)- and (L)-isomers, tautomers, atropisomers, conformers and mixtures thereof (such as racemates, mixtures of diastereomers). The substituents in the compounds of this patent application may have additional asymmetric atoms. All of these stereoisomers and their mixtures are included within the scope of this patent application. Optically active (-)- and (+)-isomers, (R)- and (S)-enantiomers and (D)- and (L)-isomers may be prepared by chiral synthesis, chiral reagents or other conventional techniques. An isomer of a compound of the present patent application can be prepared by asymmetric synthesis or chiral auxiliary, or, when the molecule contains a basic functional group (such as amino group) or an acidic functional group (such as carboxyl group), a diastereoisomer salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art to obtain pure isomers. In addition, the separation of enantiomers and diastereomers is usually completed by chromatography.

[0086] In the chemical structure of the compound described in this patent application, the bond " / " represents an unspecified configuration, that is, if chiral isomers exist in the chemical structure, the bond " / " can be or or contain both and configurations.

[0087] The compound of this patent application can exist in different tautomeric forms, and all such forms are included in the scope of this patent application. The term "tautomer" or "tautomeric form" refers to a structural isomer that exists in equilibrium and is easily converted from one isomeric form to another isomeric form. It includes all possible tautomers, i.e., exists in the form of a single isomer or in the form of a mixture of any ratio of the tautomer. Non-limiting examples include: keto-enol, imine-enamine, lactam-lactimide, etc. All tautomeric forms are within the scope of this patent application, and the naming of the compound does not exclude any tautomer.

[0088] In the present invention, a pharmaceutical composition refers to a mixture containing one or more compounds described herein or their physiologically / pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically / pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

[0089] In the present invention, pharmaceutically acceptable salts refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.

[0090] With respect to a drug or pharmacologically active agent, the term "therapeutically effective amount" refers to an amount of the drug or agent sufficient to achieve or at least partially achieve the desired effect. The determination of a therapeutically effective amount varies from person to person, depending on the age and general condition of the recipient and on the specific active substance, and the appropriate therapeutically effective amount in an individual case can be determined by a person skilled in the art based on routine experiments.

[0091] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms that are, within the scope of sound medical judgment, suitable for use in contact with patient tissues without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio, and effective for the intended use.

[0092] As used herein, the singular form of "a," "an," and "the" include plural references and vice versa unless the context clearly dictates otherwise.

[0093] In the present invention, "plurality" refers to 2 or more, for example, it can be an integer of 2, 3, 4, 5, 6, 7, 8, etc.

[0094] When the term "about" is applied to a parameter such as pH, concentration, temperature, etc., it indicates that the parameter can vary by ±10%, and sometimes more preferably within ±5%. As will be understood by those skilled in the art, when a parameter is not critical, numbers are generally given only for illustrative purposes and not for limitation. DETAILED DESCRIPTION

[0095] In order to better explain the technical solution of the present invention, the present invention provides synthesis or biological examples of some compounds, which are not intended to further limit the protection scope of the invention.

[0096] Materials and methods

[0097] The compounds provided herein can be prepared from readily available starting materials using the following general methods and processes. It should be understood that other process conditions may also be used under given typical or preferred process conditions (i.e., reaction temperature, time, molar ratio of reactants, solvent, pressure, etc.), unless otherwise stated. Optimal reaction conditions may vary with the specific reactants or solvents used, but such conditions may be determined by conventional optimization by those skilled in the art.

[0098] Additionally, it will be apparent to those skilled in the art that conventional protecting groups may be necessary to prevent certain functional groups from undergoing undesired reactions. The selection of suitable protecting groups for particular functional groups as well as suitable conditions for protection and deprotection are well known in the art.

[0099] The compounds provided herein can be separated and purified by known standard methods. Such methods include, but are not limited to, recrystallization, column chromatography, HPLC or supercritical fluid chromatography (SFC). The following reaction schemes are provided for the preparation details of the representative pyrazoles listed herein. The compounds provided herein can be prepared by those skilled in the art of organic synthesis from known or commercially available raw materials and reagents.

[0100] The compounds of the present invention can be prepared according to methods described in the art and using appropriate reagents, starting materials and purification methods known to those skilled in the art.

[0101] Example 1 Synthesis of Compound KH001

[0102]

[0103]

[0104] Compound KHB-190: Potassium thiocyanate (8.2 g, 84.5 mmol, 1.2 eq.) was added to an EtOH solution (200.0 mL) of compound MeI (10.0 g, 70.5 mmol, 1.0 eq.) and stirred at 40°C for 6 h. Then, zinc chloride (14.4 g, 105.7 mmol, 1.5 eq.) and sodium azide (6.9 g, 105.7 mmol, 1.5 eq.) were added to the reaction system and reacted at 50°C for 2 h. After the reaction was complete, the reaction solvent was removed under reduced pressure, the crude product was diluted with 500 mL of 5% w / w sodium hydroxide aqueous solution, filtered, the filter cake was washed with 100 mL of 5% w / w sodium hydroxide aqueous solution, the filtrate was adjusted to pH 1 with concentrated hydrochloric acid, extracted with ethyl acetate (100 mL x 3), the organic phases were combined and washed once with 200 mL of saturated brine, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain compound KHB-190. 1 H NMR(400MHz,DMSO)δ2.71(s,3H).LC-MS(ESI,m / z):117.2[M+1] + .

[0105] Compound KHB-191: Cesium carbonate (7.3 g, 22.4 mmol, 2.0 eq.) and BnBr (2.3 g, 13.4 mmol, 1.2 eq.) were added (dropwise) to compound KHB-190 (1.3 g, 11.2 mmol, 1.0 eq.) in DMF (105 mL), and the mixture was stirred at room temperature for 16 h. After the reaction was complete, water (50 mL) was added to dilute the mixture, and the mixture was extracted with ethyl acetate (50 mL*3). The organic phases were combined, washed with saturated brine (100 mL), concentrated under reduced pressure, and flash purified to obtain compound KHB-191. 1H NMR(400MHz, DMSO)δ7.43-7.35(m,3H),7.26(d,J=7.7Hz,2H),5.56(s,2H),2.72(s,3H).LC-MS(ESI,m / z):207.1[M+1] + .

[0106] Compound KHB-192: m-CPBA (2.1 g, 12.3 mmol, 3.0 eq.) was added to a DCM (10 mL) solution of compound KHB-191 (850 mg, 4.1 mmol, 1.0 eq.), and the mixture was stirred at room temperature for 16 h. After the reaction was complete, the mixture was filtered under reduced pressure, the filter cake was washed with DCM (20 mL*2), the filtrate was dried under reduced pressure, and the residue was diluted with a saturated aqueous sodium bicarbonate solution (15 mL), extracted with ethyl acetate (15 mL*3), the organic phases were combined and washed once with saturated brine (15 mL), dried over anhydrous sodium sulfate, filtered, the organic phase was dried, and flash purified to obtain compound KHB-192. 1 H NMR(400MHz, DMSO)δ7.49-7.38(m,5H),6.11(s,2H),3.54(s,3H).LC-MS(ESI,m / z):239.1[M+1] + ,279.1[M+41] + .

[0107] Compound KHB-193: 3-bromophenol (300.0 mg, 1.3 mmol) and Cs2CO3 (820.5 mg, 2.5 mmol) were added to a solution of compound KHB-192 (261.4 mg, 1.5 mmol) in DMSO (6 mL), and the mixture was heated and stirred at 80°C for 10 hours. After the reaction is complete, dilute with water (20 mL), extract three times with ethyl acetate (20 mL), combine the organic phases and wash once with saturated brine (20 mL), dry over anhydrous sodium sulfate, filter, and shrink to dryness. The crude product is purified by flash to obtain compound KHB-193. 1H NMR (400 MHz, DMSO) δ7.75 (dd, J=2.3, 1.4 Hz, 1H), 7.59-7.53 (m, 1H), 7.47 (td, J=4.7, 2.3 Hz, 2H), 7.43-7.34 (m, 5H), 5.57 (s, 2H); LC-MS (ESI, m / z): 331.0 [M+1]+.

[0108] Compound KHB-194: To a solution of compound KHB-193 (250.0 mg, 0.75 mmol) in dioxane (10 mL), B2(pin)2 (384.3 mg, 1.5 mmol), Pd(dppf)Cl2 (55.2 mg, 0.075 mmol) and potassium acetate (222.3 mg, 2.3 mmol) were added and the mixture was stirred at 80 °C under nitrogen atmosphere for 3 h. The reaction system was directly used for the next step.

[0109] Compound KHB-195: Water (2 mL), compound KHB-23 (275.7 mg, 0.75 mmol, 1.0 eq), Pd(dppf)Cl2 (55.2 mg, 0.075 mmol), and K2CO3 (208.7 mg, 1.5 mmol) were added to the reaction solution of the previous step, and the mixture was stirred at 80°C for 3 h under nitrogen atmosphere. The reaction solvent was removed under reduced pressure, and the crude product was directly flash purified to obtain compound KHB-195. LC-MS (ESI, m / z): 519.2 [M-OH]+.

[0110] Compound KH001: 370 mg Pd(OH)2 / C (Pd 20% w / w, H2O 50% w / w) was added to a solution of compound KHB-195 (370 mg, 0.69 mmol) in methanol (8 mL), and the mixture was stirred at room temperature for 16 h under a hydrogen atmosphere. The reaction solution was filtered, the filtrate was dried and dissolved in 2 mL DMF / H2O (3 / 1) and purified by Pre-HPLC to obtain compound KH001. NMR (400MHz, MeOD) δ7.81-7.68(m,2H),7.61(t,J=7.8Hz,1H),7.36(d,J=11.7Hz,3H),7. 20 (d, J=8.4Hz, 1H), 7.08 (d, J=8.4Hz, 1H), 1.58 (s, 6H); LC-MS (ESI, m / z): 429.1[M-OH]+.

[0111] Example 2 Synthesis of Compounds KH002 and KH003

[0112]

[0113] Compound KHB-196: To a mixed solution of compound KHB-23 (200 mg, 547.7 umol, 1.0 eq) in 1,4-dioxane / H2O=5 / 1 (18 mL) were added KHB-121 (144.0 mg, 657.2 umol, 1.2 eq), Pd(dppf)Cl2 (40.0 mg, 54.8 umol, 0.1 eq), and cesium carbonate (446.1 mg, 1.4 mmol, 2.5 eq) under nitrogen atmosphere and stirred at 85°C for 3 h. After the reaction was complete, the solvent was removed under reduced pressure and flash purified to obtain compound KHB-196. 1H NMR (400MHz, DMSO) δ11.35 (s, 1H), 8.17 (s, 1H), 7.91-7.82 (m, 2H), 7.56 (s, 1H), 7.22 (t, J = 7.7 Hz, 1H), 7.02 (dt, J = 8.5, 4.9 Hz, 2H), 6.73 (d, J = 8.0 Hz, 1H), 6.67 (s, 1H), 6.60 (d, J = 7.4 Hz, 1H), 5.25 (s, 2H), 4.98 (s, 1H), 1.44 (s, 6H). LC-MS (ESI, m / z): 360.2 [M-17]+.

[0114] Compounds KH002 and KH003: To a solution of compound KHB-196 (40 mg, 106.0 umol, 1.0 eq) in EtOH (4.0 mL) were added 2,4,6-trifluoropyrimidine (28.4 mg, 212.0 umol, 2.0 eq) and sodium carbonate (22.5 mg, 212.0 umol, 2.0 eq) and the reaction was stirred at room temperature for 16 h. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was diluted with water (20 mL), extracted with ethyl acetate (10 mL*3), the organic phases were combined and washed once with saturated brine (10 mL), dried over anhydrous sodium sulfate, filtered, dried under reduced pressure, dissolved in DMF / H2O=1 / 1 (2 mL), and purified by Pre-HPLC to obtain compound KH002. 1HNMR (400 MHz, DMSO) δ11.45 (s, 1H), 10.55 (s, 1H), 8.22 (s, 1H), 7.98-7.86 (m, 2H ),7.81(d,J=7.9Hz,1H),7.70-7.57(m,3H),7.31(d,J=7.5Hz,1H),7.00(q,J=8.5Hz,2H),6.38(d,J=1.8Hz,1H),5.00 (s,1H),1.43(s,6H).;19FNMR(376MHz,DMSO)δ-44.23(s),-65.26(s),-130.86(s).;LC-MS(ESI,m / z): 474.0[M-17]+.

[0115] Compound KH003: (7.1 mg, 13.6%) 1H NMR (400 MHz, DMSO) δ 11.44 (s, 1H), 10.54 (s, 1H), 8.21 (brs, 1H), 7.99-7.83 (m, 3H), 7.74 (s, 1H), 7.60 (d, J = 7.7 Hz, 2H), 7.22 (d, J = 6.4 Hz, 1H), 6.98 (d, J = 11.4 Hz, 2H), 6.52 (s, 1H), 4.99 (s, 1H), 1.43 (s, 6H); 19F NMR (376 MHz, DMSO) δ -57.08 (s), -130.69 (s); LC-MS (ESI, m / z): 474.9 [M-17]+.

[0116] Example 3 Synthesis of Compound KH004

[0117]

[0118] Compound KHB-199: EDCI (3.4 g, 17.6 mmol, 1.5 eq) and HOBT (2.4 g, 17.6 mmol, 1.5 eq) were added to a dichloromethane (20 mL) solution of compound KHB-197 (2.0 g, 11.8 mmol, 1.0 eq), and the mixture was stirred at room temperature for 30 min. KHB-198 (2.6 g, 14.1 mmol, 1.2 eq) was then added and stirred at room temperature for 2 h. After the reaction was complete, 30 mL of water was added to the reaction system and extracted three times with 30 mL of dichloromethane. The organic phases were combined, washed once with 30 mL of 15% w / w sodium chloride aqueous solution, dried over anhydrous sodium sulfate, filtered, dried, and flash purified to obtain compound KHB-199. NMR(400MHz,DMSO)δ8.85(t,J=6.0Hz,1H),7.54(s,1H),7.45-7.40(m,2H),7.36-7.27(m,3H),7 .18 (dd, J=9.1, 4.3Hz, 1H), 4.49 (d, J=6.2Hz, 2H), 3.90 (s, 3H).; LC-MS (ESI, m / z): 337.9[M+H]+.

[0119] Compound KHB-200: Add B2(pin)2 (450.5 mg, 1.8 mmol, 2.0 eq) and Pd(dppf)Cl2 (64.9 mg, 88.7 umol, 0.1 eq) and AcOK (261.2 mg, 2.7 mmol, 3.0 eq) to a solution of compound KHB-199 (300.0 mg, 887.1 umol, 1.0 eq) in dioxane (10 mL) and stir at 80 °C for 2 h under nitrogen atmosphere. Remove the reaction solvent under reduced pressure and flash purify the crude product to obtain compound KHB-200. NMR (400MHz, DMSO) δ8.80(t,J=6.0Hz,1H),7.70(s,1H),7.55(d,J=7.2Hz,1H),7.46(dd,J=9.1,3.2Hz,2H),7.38-7.28 (m,2H),7.17(dd,J=9.1,4.3Hz,1H),4.49(d,J=6.1Hz,2H),3.90(s,3H),1.29(s,12H).;LC-MS(ESI,m / z):386.2[M+H] + .

[0120] Compound KH004: Compound KHB-200 (31.7 mg, 82.2 umol, 1.2 eq.) and potassium carbonate (18.9 mg, 136.9 umol, 2.0 eq.) were added to a mixed solution of compound KHB-23 (25.0 mg, 68.5 umol, 1.0 eq.) in dioxane / water (5 / 1) (5 mL) and stirred at 85°C for 2 h. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was dissolved in methanol, and palladium black was removed by flash column chromatography. The fractions were condensed to dryness, diluted with 2 mL of DMSO, and purified by Pre-HPLC to obtain compound KH004. NMR (400MHz, DMSO) δ8.80(t,J=6.0Hz,1H),7.70(s,1H),7.55(d,J=7.2Hz,1H),7.46(dd,J=9.1,3.2Hz,2H),7.38-7.28 (m,2H),7.17(dd,J=9.1,4.3Hz,1H),4.49(d,J=6.1Hz,2H),3.90(s,3H),1.29(s,12H).;LC-MS(ESI,m / z):526.2[M-OH] + .

[0121] Example 4 Synthesis of Compound KH005

[0122]

[0123] Compound KHB-201: Under nitrogen protection, 1,3-dibromobenzene (5.9 g, 25.37 mmol), Pd2(dba)3 (1.94 g, 2.11 mmol), 4,5-bis(diphenylphosphine)-9,9-dimethyloxanthene (1.22 g, 2.11 mmol) and cesium carbonate (17.23 g, 525.85 mmol) were added to ultra-dry dioxane (200 mL) of compound KHB-131 (3.7 g, 21.14 mmol) and the system was heated to 105°C and stirred for 4 hours. After the reaction is complete, the mixture is extracted with ethyl acetate, washed twice with water, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure. The organic phase is concentrated, and the residue is loaded on silica gel and purified by column chromatography to obtain compound KHB-201. 1H NMR (400MHz, DMSO-d6) δ10.09 (s, 1H), 7.72 (t, J = 1.9 Hz, 1H), 7.47-7.42 (m, 1H), 7.42-7.34 (m, 5H), 7.23 (t, J = 8.1 Hz, 1H), 7.08-7.03 (m, 1H), 5.83 (s, 2H); LC-MS (ESI, m / z): 331.2 [M+1] + .

[0124] Compound KHB-202: Under nitrogen protection, to the ultra-dry dioxane (50 mL) of compound KHB-201 (3 g, 9.09 mmol) were added bis(2-methyl-2,4-pentanediol) borate (3.46 g, 13.64 mmol), potassium acetate (2.67 g, 27.27 mmol) and [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (665 mg, 0.91 mmol). The system was heated to 85 degrees and stirred for 4 hours. After the reaction is complete, the mixture is extracted with ethyl acetate, washed twice with water, dried over anhydrous sodium sulfate, and the solvent is removed under reduced pressure. The organic phase is concentrated, and the residue is loaded on silica gel and purified by column chromatography to obtain compound KHB-202. 1H NMR (400MHz, DMSO-d6) δ9.81 (s, 1H), 7.84 (d, J = 2.1 Hz, 1H), 7.73-7.68 (m, 1H), 7.47-7.41 (m, 5H), 7.37-7.32 (m, 1H), 7.25 (d, J = 7.2 Hz, 1H), 5.86 (s, 2H), 1.35 (s, 12H); LC-MS (ESI, m / z): 378.4 [M+1] + .

[0125] Compound KHB-203: Under nitrogen protection, compound KHB-195 (667 mg, 1.77 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (117 mg, 0.16 mmol) and cesium carbonate (1314 mg, 4.03 mmol) were added to a solution of compound KHB-26 (500 mg, 1.61 mmol) in dioxane: water = 16 ml: 4 ml. The system was heated to 85 degrees and stirred for 3 hours. After the reaction was complete, the mixture was extracted with ethyl acetate, washed twice with water, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain compound KHB-203. LC-MS (ESI, m / z): 482.2 [M+1] + .

[0126] Compound KH005: Under hydrogen protection, 10% palladium on carbon (880 mg, 0.83 mmol) was added to a methanol solution (10 mL) of compound KHB-203 (300 mg, 0.83 mmol), and the system was heated to room temperature and stirred for reaction overnight. After the reaction was complete, palladium carbon was removed by filtration, the filtrate was concentrated, the crude product was dissolved in 2 ml of DMF, and purified by reverse phase preparative chromatography to obtain compound KH005. 1H NMR (400 MHz, DMSO) δ 10.84 (s, 1H), 9.85 (s, 1H), 8.11 (brs, 1H), 7.65-7.31 (m, 5H), 6.92 (d, J = 7.4 Hz, 1H), 2.73 (brs, 2H), 2.73 (brs, 2H), 1.77-1.43 (m, 4H). LCMS (ESI, m / z): 392.1 [M+1] + .

[0127] Example 5 Synthesis of Compound KH006

[0128]

[0129] Compound KHB-204: KHB-202 (826.4 mg, 2.2 mmol, 1.0 eq) and Pd(dppf)Cl2 (160.2 mg, 219.1 umol, 0.1 eq) and K2CO3 (605.5 mg, 4.4 mmol, 2.0 eq) were added to a mixed solution of compound KHB-23 (800.0 mg, 2.2 mmol, 1.0 eq) in dioxane (15 mL) and water (3 mL), and the mixture was stirred at 80°C for 3 h. The reaction solvent was removed under reduced pressure, and the resulting crude product was flash purified to obtain compound KHB-204, which was used directly in the next step. LC-MS (ESI, m / z), 518.2 [M-OH]+.

[0130] Compound KH006: To a mixed solution of compound KHB-204 (600.0 mg, 1.1 mmol, 1.0 eq.) in methanol (8 mL) and tetrahydrofuran (2 mL) was added 600 mg Pd(OH)2 (Pd 20% w / w, H2O 50% w / w) and stirred at room temperature for 16 hours. The reaction solution was filtered, the filtrate was concentrated to dryness and washed with 4 mL Dissolved in DMF / H2O (3 / 1) and purified by Pre-HPLC to obtain compound KH006. 1HNMR (400 MHz, DMSO) δ 11.42 (s, 1H), 9.86 (s, 1H), 8.22 (s, 1H), 7.92 (t, J = 10.9 Hz, 1H), 7.87 (s, 1H), 7.76 (d, J = 8.1 Hz, 1H), 7.69 (s, 1H), 7.60 (s, 1H), 7.53 (t, J = 7.9 Hz, 1H), 7.04 (d, J = 7.4 Hz, 1H), 7.02-6.95 (m, 2H), 4.99 (s, 1H), 1.43 (s, 6H). LC-MS (ESI, m / z): 428.1 [M-OH] + .

[0131] Example 6 Synthesis of Compound KH007

[0132]

[0133] Compound KHB-205: Under nitrogen protection, compound KHB-202 (675 mg, 1.79 mmol), [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium (117 mg, 0.16 mmol) and cesium carbonate (1330 mg, 4.08 mmol) were added to a solution of compound KHB-27 (500 mg, 1.63 mmol) in dioxane and water = 16 ml:4 ml. The system was heated to 85°C and stirred for 3 hours. After the reaction was complete, the mixture was extracted with ethyl acetate, washed twice with water, dried over anhydrous sodium sulfate, and the solvent was removed under reduced pressure to obtain compound KHB-205. LCMS (ESI, m / z): 378.2 [M+1] + .

[0134] Compound KH007: Under hydrogen protection, 10% palladium carbon (2004 mg, 1.89 mmol) was added to a methanol solution (20 mL) of compound KHB-205 (300 mg, 0.63 mmol), and the system was stirred overnight at room temperature. After the reaction was complete, the palladium carbon was filtered off, the filtrate was concentrated, the crude product was dissolved in 2 ml of DMF, and purified by reverse phase preparative chromatography to obtain compound KH007. 1HNMR (400MHz, DMSO) δ11.55(s,1H),9.89(s,1H),8.25(s,1H),7.98(d,J=10.9Hz,1H),7.75(d,J=7.6Hz,2H),7.70(s,1H),7.65( brs,1H),7.54(t,J=7.9Hz,1H),7.33(t,J=7.7Hz,1H),7.07(t,J=6.2Hz,2H),6.89(t,J=7.6Hz,1H).LCMS(ESI,m / z): 388.2[M+1] + .

[0135] Example 7 Synthesis of Compounds KH008 and KH009

[0136]

[0137] Compound KHB-207: Compound KHB-121 (1.2 g, 5.47 mmol), Pd(dppf)Cl2 (334 mg, 0.46 mmol) and cesium carbonate (3.714 g, 11.40 mmol) were added to a mixed solution of compound KHB-27 (1.4 g, 4.56 mmol) in 1,4-dioxane (30 mL) and water (7.5 mL). ℃ The reaction was carried out for 3 hours. After the reaction was complete, water was added after the reaction solution returned to room temperature, and ethyl acetate was added to extract the organic phase, which was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under vacuum to obtain a crude product; the crude product was then purified by silica gel column chromatography to obtain compound KHB-207. 1 H NMR (400MHz, DMSO) δ11.47(s,1H),8.20(s,1H),7.92(d,J=10.9Hz,1H),7.72(d,J=8.2Hz,1H),7.60(s,1H),7.36-7.29(m,1H),7.23(t,J=7.7Hz,1H ),7.14(d,J=8.0Hz,1H),6.90(t,J=7.2Hz,1H),6.73(d,J=7.3Hz,1H),6.6 8(s,1H),6.60(d,J=7.4Hz,1H),5.25(s,2H); LCMS(ESI,m / z):320.1[M+1] + .

[0138] Compounds KH008 and KH009: Add 2,4,6-trifluoropyrimidine (252 mg, 1.88 mmol) and sodium carbonate (200 mg, 1.88 mmol) to a solution of compound KHB-207 (300 mg, 0.94 mmol) in ethanol (6 mL), and stir at room temperature to react overnight. After the reaction is complete, vacuum concentrate to obtain a crude product; the crude product is dissolved in a solution of dimethyl sulfoxide and tetrahydrofuran (1:1), and the compound KH008 is prepared and purified by reverse phase preparation. LCMS (ESI, m / z): 434.1 [M+1] + . 1 H NMR (400MHz, DMSO) δ11.57(s,1H),10.56(s,1H),8.25(s,1H),7.98(d,J=10.9Hz,1H),7.86-7.5 6(m,5H),7.41-7.26(m,2H),7.08(d,J=8.0Hz,1H),6.91(t,J=7.5Hz,1H),6.38(d,J=2.2Hz,1H).

[0139] Compound KH009: LCMS (ESI, m / z): 434.1 [M+1] + . 1 H NMR (400MHz, DMSO) δ11.55(s,1H),10.53(s,1H),8.24(s,1H),8.02-7.86(m,2H),7.78-7.55(m,4H),7 .34(t,J=7.7Hz,1H), 7.22(d,J=7.5Hz,1H), 7.06(d,J=7.9Hz,1H), 6.90(t,J=7.6Hz,1H), 6.51(s,1H).

[0140] Example 8 Synthesis of Compound KH010

[0141]

[0142] Compound KHB-208: Compound 2,4,6-trichloro-1,3,5-triazine (347 mg, 1.88 mmol) and potassium carbonate (780 mg, 5.64 mmol) were added to a solution of compound KHB-207 (600 mg, 1.88 mmol) in acetonitrile (15 mL), and the mixture was reacted at room temperature for 3 hours. After the reaction was complete, water was added to the reaction solution, and the organic phase was extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain compound KHB-208.

[0143] Compound KH010: Potassium acetate (378 mg, 3.85 mmol) was added to a mixed solution of compound KHB-208 (600 mg, 1.28 mmol) in acetic acid (6 mL) and water (3 mL), and the mixture was reacted at 90°C overnight. After the reaction was complete, the mixture was concentrated under vacuum to obtain a crude product; the crude product was dissolved in a solution of dimethyl sulfoxide and tetrahydrofuran (1:1), and the compound KH010 was prepared and purified by reverse phase. LCMS (ESI, m / z): 431.1 [M+1] + . 1 H NMR (400MHz, DMSO) δ11.56(s,1H),10.75(s,1H),10.66(brs,1H),9.27(s,1H),8.25(s,1H),7.98(d,J=10.9H z,1H),7.83-7.72(m,3H),7.70-7.56(m,2H),7.38-7.25(m,2H),7.04(d,J=8.0Hz,1H),6.91(t,J=7.5Hz,1H).

[0144] Example 9 Synthesis of Compounds KH011 and KH012

[0145]

[0146] Compound KHB233: Compound KHB128 (500 mg, 1.28 mmol), compound KHB27 (380 mg, 1.28 mmol), Pd(dppf)Cl2 (119 mg, 0.13 mmol), K2CO3 (675 mg, 3.86 mmol) were dissolved in dioxane (5 mL) and water (1 mL), and the reaction was stirred at 90°C for 3 hours under N2 protection. After the reaction was completed, the reaction system was diluted with 10 mL of water, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain compound KHB233. LCMS (ESI, m / z): 334.1 [M+H] + .

[0147] KH011, KH012: Add compound 2, -4,6-trifluoropyrimidine (66 mg, 0.50 mmol) and DIEA (85 mg, 0.66 mmol) to a DMSO (2 mL) solution of compound KHB233 (110 mg, 0.33 mmol), and stir the reaction at 70°C for 16 hours. After the reaction is completed, dilute the reaction system with 5 mL of water, extract with ethyl acetate (5 mL x 3), combine the organic phases, wash with saturated brine (5 mL), dry and concentrate with anhydrous sodium sulfate, pass through a C18 reverse phase column, and prepare with ammonium bicarbonate to obtain compound KH011: LCMS (3.1 mg) (ESI, m / z): 448.3 [M+H] +, 1 H NMR (400MHz, MeOD) δ7.83 (d, J = 10.3 Hz, 1H), 7.53 (m, 3H), 7.36 (m, 2H), 6.91 (d, J = 4.1 Hz, 2H), 5.99 (s, 1H), 1.96 (s, 3H).

[0148] Compound KH012: LCMS (15.73 mg) (ESI, m / z): 448.3 [M+H] + . 1 H NMR (400MHz, DMSO) δ11.53(s,1H),9.94(s,1H),8.25(s,1H),7.98(d,J=10.8Hz,1H),7.73(d,J=8.2Hz,1H),7.65(s,1H),7.52(d,J =7.5Hz,1H),7.44(t,J=7.9Hz,1H),7.34(d,J=4.1Hz,1H),7.26(d,J=7.5Hz,1H),6.89(d,J=3.5Hz,2H),6.38(s,1H),1.88(s,3H).

[0149] Example 10 Synthesis of Compound KH013

[0150]

[0151]

[0152] Compound KHB234: Add B2(pin)2 (1.24 g, 4.88 mmol), potassium acetate (639 mg, 6.51 mmol) and Pd(dppf)Cl2 (236 mg, 0.33 mmol) to a solution of compound KHB27 (1 g, 3.26 mmol) in dioxane (12 mL) and react at 90°C for 3 h. After the reaction is complete, it is directly used for the next step. LCMS (ESI, m / z): 355.1 [M+H] +.

[0153] Compound KHB-236: KHB-215 (200 mg, 2.02 mmol), KHB-214 (58 mg, 0.40 mmol), t-BuOK (680 mg, 6.06 mmol) and CuI (39 mg, 0.20 mmol) were added to a solution of compound KHB-235 (600 mg, 2.02 mmol) in t-BuOH (8 mL). The reaction was stirred at 80 ° C for 3 hours under N2 protection. After the reaction was completed, the reaction system was diluted with 10 mL of water, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and the crude product was purified by silica gel column chromatography to obtain compound KHB-236. LCMS (ESI, m / z): 270.0 [M+H] + .

[0154] Compound KH013: Compound KHB236 (204 mg, 0.80 mmol), KHB-234 (450 mg, 1.20 mmol), Pd(dppf)Cl2 (139 mg, 0.23 mmol), Cs2CO3 (1240 mg, 3.60 mmol) were dissolved in Dioxane (5 mL) and water (1 mL), and the reaction was stirred at 90°C for 3 hours under N2 protection. After the reaction was completed, the reaction system was diluted with 10 mL of water, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried and concentrated over anhydrous sodium sulfate, and purified by C18 reverse phase column to obtain compound KH013. LCMS (ESI, m / z): 416.2 [M+H] + . 1 H NMR (400MHz, DMSO) δ11.53(s,1H),8.86(s,1H),8.25(s,1H),7.98(d,J=10.6Hz,1H),7.83(d,J=8.2Hz,1H),7.74 (d,J=8.2Hz,1H),7.66(s,1H),7.36(m,2H),7.02(d,J=7.4Hz,1H),6.95-6.76(m,2H),4.26(s,3H),1.97(s,3H).

[0155] Example 11 Synthesis of Compound KH014

[0156]

[0157] Compound KHB238: Compound a (1.24 g, 4.88 mmol), potassium acetate (639 mg, 6.51 mmol) and Pd(dppf)Cl2 (236 mg, 0.33 mmol) were added to a solution of compound KHB237 (synthesized by the same method as KHB-27, 1 g, 3.26 mmol) in dioxane (10 mL), and the mixture was reacted at 90°C for 3 h. After the reaction was complete, it was directly used for the next step. LCMS (ESI, m / z): 355.1 [M+H] + .

[0158] Compound KH014: Compound KHB236 (60 mg, 0.22 mmol), potassium carbonate (62 mg, 0.45 mmol) and Pd(dppf)Cl2 (16 mg, 0.022 mmol) were added to a solution of compound KHB238 (79 mg, 0.22 mmol) in dioxane (3 mL), and the mixture was reacted at 90°C for 3 h. After the reaction was complete, water was added to the reaction solution, and ethyl acetate was added to extract the organic phase, which was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was then purified by reverse phase preparative chromatography to obtain compound KH014. LCMS (ESI, m / z): 416.2 [M+H]+. 1H NMR (400MHz, DMSO) δ11.59(s,1H),8.65(s,1H),7.90(s,1H),7.78(s,1H),7.69(d,J=8.2Hz,1H),7.62(d,J=7.3Hz, 1H),7.40(t,J=7.8Hz,1H),7.34-7.27(m,1H),7.13(d,J=6.7Hz,1H),6.95-6.85(m,3H),3.92(s,3H),1.93(s,3H).

[0159] Example 12 Synthesis of Compound KH015

[0160]

[0161] Compound KHB249: Compound KHB-127 (236 mg, 0.77 mmol), potassium carbonate (213 mg, 1.54 mmol) and Pd(dppf)Cl2 (56 mg, 0.077 mmol) were added to a dioxane (5 mL) solution of compound KHB27 (180 mg, 0.77 mmol) and reacted at 90°C for 3 h. After the reaction was complete, water was added to the reaction solution, and ethyl acetate was added to extract the organic phase, which was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was then purified by silica gel column chromatography to obtain compound KHB249. LCMS (ESI, m / z): 376.1 [M+H+ACN]+ . 1 H NMR (400MHz, DMSO) δ11.48(s,1H),8.22(s,1H),7.95(d,J=10.6Hz,1H),7.72(d,J=8.2Hz,1H),7.62(s,1H),7.31(t,J=7.6 Hz,1H),7.19(t,J=7.7Hz,1H),7.00(d,J=8.1Hz,1H),6.87(t,J=7.5Hz,1H),6.80-6.73(m,2H),3.32(s,3H),1.81(s,3H).

[0162] Compound KH015: Compound KHB211-1 (99 mg, 0.61 mmol) and potassium carbonate (99 mg, 0.72 mmol) were added to a solution of compound KHB249 (120 mg, 0.36 mmol) in acetonitrile (3 mL), and the mixture was reacted at 80°C for 2 h. After the reaction was complete, water was added to the reaction solution, and ethyl acetate was added to extract the organic phase, which was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a crude product, which was then purified by reverse phase preparative chromatography to obtain compound KH015. LCMS (ESI, m / z): 417.1 [M+H] + . 1 HNMR (400MHz, DMSO) δ11.58(s,1H),8.27(s,1H),8.01(d,J=10.7Hz,1H),7.75(d,J=8.2Hz,1H),7.66(d,J=8.2Hz,2H ),7.55(t,J=7.9Hz,1H),7.42-7.32(m,2H),6.92(t,J=7.5Hz,1H),6.82(d,J=7.9Hz,1H),4.00(s,3H),1.93(s,3H).

[0163] Example 13 Synthesis of Compound KH016

[0164]

[0165] Compound KHB225: Compound KHB235 (1.77 mg, 5.96 mmol), Brettphos-Pd-G3 (540 mg, 0.60 mmol) and potassium carbonate (1.65 g, 11.90 mmol) were added to tert-butyl alcohol (20 mL) of compound KHB254 (500 mg, 5.96 mmol) in sequence, and stirred at 80°C for 6 h under nitrogen protection. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound KHB225. LCMS (ESI, m / z): 253.3 / 255.3 [M+H] + . 1 HNMR (400MHz, DMSO) δ9.27(s,1H),7.77(d,J=8.0Hz,1H),7.63(s,1H),7.31(d,J=7.8Hz,1H),7.11(t,J=8.0Hz,1H),6.92(s,1H),2.32(s,3H).

[0166] Compound KH016: Compound KHB234 (137.0 mg, 0.386 mmol), Pd(dtbpf)Cl2 (20 mg, 0.026 mmol) and potassium carbonate (107.0 mg, 0.77 mmol) were added to a mixed solution of compound KHB225 (65 mg, 0.257 mmol) in 1,4-dioxane (5 mL) and water (1 mL) in sequence, and stirred at 90°C for 6 h under nitrogen protection. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was diluted with water, extracted with ethyl acetate, and the organic phase was washed twice with water and twice with saturated brine, dried over anhydrous sodium sulfate, concentrated to obtain a crude product, and then purified by reverse phase preparation to obtain compound KH016. LCMS (ESI, m / z): 401.2 [M+H] + . 1 H NMR (400MHz, DMSO) δ11.51(s,1H),9.20(s,1H),8.24(s,1H),8.04-7.96(m,2H),7.73(d,J=8.2Hz,1H),7.64(s,2H) ,7.38-7.31(m,2H),7.04(d,J=7.4Hz,1H),6.97(s,1H),6.88(t,J=7.6Hz,1H),6.78(d,J=8.0Hz,1H),1.95(s,3H).

[0167] Example 14 Synthesis of Compound KH017

[0168]

[0169] Compound KHB257: Compound KHB170 (2.5 g, 25.1 mmol) and MgSO4 (3.02 g, 25.1 mmol) were added to a solution of compound KHB258 (5.0 g, 25.1 mmol) in ethanol (60 mL) in turn, and the mixture was stirred at 80°C for 2 h. After the reaction was complete, the solid was removed by filtration, and the reaction solvent was removed under reduced pressure to obtain a crude compound KHB257. The crude product was directly used for the next step. LCMS (ESI, m / z): 281.2 [M+H] + .

[0170] Compound KHB258: Add NaBH4 (2.92 g, 77.1 mmol) to a solution of compound KHB-257 (7.2 g, 25.7 mmol) in ethanol (60 ml). Stir the reaction at 50°C for 2 h. After the reaction is complete, add saturated aqueous ammonium chloride to quench the reaction solution, dilute with water, extract with ethyl acetate, wash the organic phase with water twice, dry with anhydrous sodium sulfate, and concentrate to obtain a crude product. The crude product is purified by silica gel column chromatography to obtain compound KHB258. LCMS (ESI, m / z): 383.3 [M+H] + . 1 H NMR (400MHz, DMSO) δ6.62 (d, J = 7.2Hz, 1H), 3.75-3.73 (m, 1H), 3.61-3.56 (m, 1H), 2.97- 2.75(m,2H),2.61(s,1H),1.91(d,J=5.4Hz,1H),1.83-1.46(m,3H),1.37-1.20(m,9H).

[0171] Compound KHB259: Add 3M HCl / EA (10 mL) solution to ethyl acetate (15 mL) of compound KHB258 (1.5 g, 5.3 mmol), and stir at room temperature for 1 h. After the reaction is complete, filter the reaction solution to obtain a solid, which is washed with EA and dried to obtain compound KHB259. LCMS (ESI, m / z): 183.2 [M+H] + . 1 H NMR(400MHz,DMSO)δ9.47(s,2H),4.11(s,3H),3.82-3.72(m,1H),3.30-3.27(m,1H),3.14-3.08(m, 1H),2.84-2.71(m,2H),2.01-1.90(m,1H),1.87-1.82(m,1H),1.78-1.67(m,1H),1.56-1.43(m,1H).

[0172] Compound KHB261: Compound KHB-260 (synthesized by the same method as KHB-28, 100 mg, 0.60 mmol), Pd2(dba)3 (23 mg, 0.024 mmol), BINAP (20 mg, 0.024 mmol) and cesium carbonate (237 mg, 0.73 mmol) were added to a solution of compound KHB259 (60 mg, 0.60 mmol) in dioxane (3 mL) in sequence, and stirred at 110°C for 12 h under nitrogen protection. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound KHB261. LCMS (ESI, m / z): 391.2 [M+1] + .

[0173] Compound KH017: 30% KOH (430 mg, 2.3 mmol) and H2O2 (520 mg, 4.6 mmol) were added to a solution of compound KHB261 (90 mg, 0.23 mmol) in dimethyl sulfoxide (10 mL) in turn, and the mixture was stirred at room temperature for 2 h. The reaction solution was filtered and concentrated under reduced pressure to obtain a crude product, which was then purified by reverse preparation to obtain compound KH017. LCMS (ESI, m / z): 409.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ11.50(s,1H),8.01(d,J=7.8Hz,1H),7.69(t,J=10.6Hz,2H),7.45(s ,1H),7.35(t,J=7.6Hz,1H),7.21(t,J=7.4Hz,1H),6.78(d,J=7.8Hz,1H),6.66(d,J=14. 4Hz,1H),4.09(s,3H),3.90(brs,1H),3.70(d,J=9.6Hz,1H),3.44(d,J=10.2Hz,1H),2.8 2-2.76(m,1H),2.63(brs,1H),2.17-2.12(m,1H),1.98-1.94(m,2H),1.54-1.51(m,1H).

[0174] Example 15 Synthesis of Compound KH018

[0175]

[0176] Compound KHB263: TEA (603 mg, 7.1 mmol) and BrCN (715 mg, 5.8 mmol) were added to a solution of compound KHB262 (1.0 g, 4.7 mmol) in dichloromethane (20 mL) in turn, and the mixture was stirred at room temperature for 10 h. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound KHB263. LCMS (ESI, m / z): 238.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ4.14 (t, J = 5.0 Hz, 1H), 3.59-3.35 (m, 5H), 3.02 (d, J = 8.6 Hz, 1H), 2.92 (s, 1H), 2.05-1.95 (m, 1H), 1.81-1.73 (m, 1H), 1.40 (s, 9H).

[0177] Compound KHB264: NH4Cl (500 mg, 9.2 mmol) and NaN3 (360 mg, 5.52 mmol) were added to a DMF (15 mL) solution of compound KHB263 (1.1 g, 4.6 mmol) in sequence, and the mixture was stirred at 110°C for 12 h. After the reaction was complete, water (20 mL) was slowly added to dilute the reaction solution, and the mixture was extracted with EA (30 mL x 2). The organic phases were combined and washed with saturated sodium chloride (60 mL), dried and concentrated with anhydrous sodium sulfate, and the crude product was purified by column chromatography to obtain compound KHB264. LCMS (ESI, m / z): 303.2 [M+Na] + ,583.4[2M+Na] + . 1 H NMR(400MHz,DMSO)δ7.90(s,1H),4.20(brs,1H),3.56-3.43(m,5H),3.21-3. 18(m,1H),3.02(brs,1H),2.13-2.08(m,1H),1.89-1.76(m,1H),1.34(s,9H).

[0178] Compound KHB265: Cs2CO3 (1.1 g, 3.38 mmol) and MeI (384 mg, 2.7 mmol) were added to a DMF (10 mL) solution of compound KHB264 (630 mg, 2.25 mmol) in sequence, and the mixture was stirred at room temperature for 2 h. After the reaction was complete, water (20 mL) was slowly added to dilute the reaction solution, and the mixture was extracted with EA (30 mL x 2). The organic phases were combined and washed with saturated sodium chloride (60 mL), dried and concentrated with anhydrous sodium sulfate, and the crude product was purified by column chromatography to obtain compound KHB265. LCMS (ESI, m / z): 295.2 [M+1] + ,317.3[M+Na] + . 1 H NMR (400MHz, DMSO) δ4.16 (s, 3H), 3.95 (d, J = 7.6Hz, 1H), 3.66-3.35 (m, 5H), 3.20 -3.13(m,1H),2.99(brs,1H),2.11-2.02(m,1H),1.85-1.78(m,1H),1.35(s,9H).

[0179] Compound KHB266: Add 3M HCl / EA (5 mL) solution to a solution of compound KHB265 (450 mg, 1.53 mmol) in ethyl acetate (10 mL), and stir at room temperature for 1 h. After the reaction is complete, filter the reaction solution to obtain a solid, which is washed with EA and dried to obtain compound KHB266. LCMS (ESI, m / z): 195.2 [M+H] + .

[0180] Compound KHB267: Compound KHB260 (220 mg, 0.77 mmol), Pd2(dba)3 (71 mg, 0.077 mmol), BINAP (48 mg, 0.077 mmol) and cesium carbonate (750 mg, 2.3 mmol) were added to a solution of compound KHB266 (150 mg, 0.77 mmol) in dioxane (10 mL) in sequence, and stirred at 110°C for 12 h under nitrogen protection. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound KHB267. LCMS (ESI, m / z): 403.2 [M+1] + .

[0181] Compound KH018: 30% KOH (935 mg, 5.0 mmol) and H2O2 (1.13 g, 10.0 mmol) were added to dimethyl sulfoxide (10 mL) of compound KHB267 (200 mg, 0.50 mmol) in sequence, and stirred at room temperature for 2 h. The reaction solution was filtered and concentrated under reduced pressure to obtain a crude product, which was purified by reverse preparation to obtain compound KH018. LCMS (ESI, m / z): 421.1 [M+H] + . 1 HNMR (400MHz, DMSO) δ11.48 (s, 1H), 7.83-7.52 (m, 3H), 7.50-7.34 (m, 1H), 7.25 (t, J = 7. 6Hz,1H),6.76(t,J=7.6Hz,1H),6.62(d,J=14.6Hz,1H),4.48-4.36(m,1H),4.19(s,3H), 3.95(d,J=10.8Hz,1H),3.81-3.76(m,1H),3.59-3.54(m,1H),3.41-3.33(m,1H),3.31-3 .29(m,1H),3.27-3.15(m,1H),2.90-2.87(m,1H),2.34-2.30(m,1H),2.12-2.00(m,1H).

[0182] Example 16 Synthesis of Compound KH019

[0183]

[0184] Compound KHB269: Compound KHB170 (2.26 g, 22.80 mmol) and MgSO4 (0.275 g, 2.28 mmol) were added to a solution of compound KHB268 (5.0 g, 22.80 mmol) in ethanol (50 mL) in sequence, and stirred at 80°C for 2 h. After the reaction was complete, the solid was removed by filtration, and the reaction solvent was removed under reduced pressure to obtain compound KHB269. The crude product was directly used for the next step. LCMS (ESI, m / z): 301.1 [M+H] + .

[0185] Compound KHB270: Add NaBH4 (2.6 g, 68.40 mmol) to a solution of compound KHB-269 (5.0 g, 22.80 mmol) in ethanol (20 ml). Stir and react at 50°C for 2 h. After the reaction is complete, remove the reaction solvent under reduced pressure to obtain a crude product, add a saturated aqueous solution of ammonium chloride to quench the crude product, dilute with water, extract with ethyl acetate, wash the organic phase with water twice, dry with anhydrous sodium sulfate, and concentrate to obtain a crude product. After reverse preparation and purification, compound KHB270 is obtained. LCMS (ESI, m / z): 303.5 [M+H] + .

[0186] Compound KHB271: Pd / C (300 mg) was added to a methanol (20 mL) solution of compound KHB270 (1.0 g, 3.311 mmol), and the mixture was stirred at room temperature for 2 h under a hydrogen atmosphere. After the reaction was complete, the reaction solution was filtered and the filtrate was removed under reduced pressure to obtain compound KHB271. LCMS (ESI, m / z): 169.1 [M+H] + . 1 H NMR (400MHz, DMSO) δ6.61 (d, J = 6.4Hz, 1H), 4.10 (s, 3H), 3.95-3.84 (m, 1H), 2.94-2.84(m,2H),2.78-2.57(m,2H),2.00-1.86(m,1H),1.62-1.55(m,1H).

[0187] Compound KHB272: Compound KHB260 (200 mg, 0.69 mmol), Pd2(dba)3 (60 mg, 0.07 mmol), BINAP (40 mg, 0.07 mmol) and cesium carbonate (670 mg, 2.08 mmol) were added to a solution of compound KHB271 (120 mg, 0.69 mmol) in dioxane (10 mL) in sequence, and the mixture was stirred at 110°C for 12 h under nitrogen protection. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was purified by silica gel column chromatography to obtain compound KHB272. LCMS (ESI, m / z): 377.1 [M+H] + . 1H NMR (400MHz, DMSO) δ12.19(s,1H),7.90(d,J=8.0Hz,1H),7.53(d,J=8.0Hz,1H),7.36(t,J=7.6Hz,1H),7.16(t,J=7.6Hz,1H),7.09(d,J=6.2 Hz,1H),6.64(d,J=13.8Hz,1H),4.29-4.21(m,1H),4.11(s,3H),3.78- 3.68(m,2H),3.52-3.46(m,2H),2.39-2.37(m,1H),2.15-2.04(m,1H).

[0188] Compound KH019: 30% KOH (560 mg, 3.0 mmol) and H2O2 (680 mg, 6.0 mmol) were added to dimethyl sulfoxide (10 mL) of compound KHB272 (110 mg, 0.3 mmol) in sequence, and stirred at room temperature for 2 h. The reaction solution was filtered and concentrated under reduced pressure to obtain a crude product, which was purified by reverse preparation to obtain compound KH019. LCMS (ESI, m / z): 395.2 [M+H] + . 1 HNMR (400MHz, DMSO) δ7.84(d,J=8.0Hz,1H),7.63(d,J=8.2Hz,1H),7.30(t,J=7.2Hz,1H),7.07(t,J=7.2Hz,1H),6.56(d,J=15.4Hz,1H),4.2 2(t,J=8.6Hz,1H),4.07(s,3H),3.87(s,1H),3.60-3.47(m,2H),3.43- 3.39(m,1H),3.35-3.29(m,1H),2.46-2.39(m,1H),2.08-2.00(m,1H).

[0189] Example 17 Synthesis of Compound KH020

[0190]

[0191] Compound KHB274: Add EDCI (920 mg, 4.8 mmol) and HOBT (814 mg, 3.84 mmol) to a solution of compound KHB273 (synthesized by the same method as KHB25, 1 g, 3.2 mmol) in THF:DCM (3:1, 10 ml), add ammonia water (1.634 g, 96 mmol) half an hour later, and stir the reaction at room temperature for 3 hours. After the reaction is completed, add 30 ml of water and stir, filter, wash twice with ethanol (2 mL), dry, and use the crude product for the next step.

[0192] Compound KHB275: Compound KHB274 (742 mg, 2.4 mmol), B2(Pin)2 (792 mg, 3.12 mmol), Pd(dppf)Cl2 (174 mg, 0.24 mmol), AcOK (705 mg, 7.2 mmol) were dissolved in dioxane, and the reaction was stirred at 90°C for 3 hours under N2 protection. After the reaction was completed, the crude product was purified by silica gel column chromatography to obtain compound KHB275, LCMS: 359.1 [M+H] + , 1 H NMR (400MHz, DMSO) δ10.78(s,1H),7.76(s,1H),7.60(s,1H),7.03(d,J=12.4Hz,1H),2.75(d,J=19.8Hz,4H),1.77(brs,4H),1.34(s,12H).

[0193] Compound KH020: Compound KHB275 (100 mg, 0.27 mmol), compound KHB236 (72 mg, 0.27 mmol), Pd(dppf)Cl2 (20 mg, 0.0271 mmol), K2CO3 (111 mg, 0.81 mmol) were dissolved in dioxane (1 ml) and water (0.2 mL). After the reaction was completed, the mixture was concentrated and purified by C18 reverse phase column to obtain compound KH020. LCMS: 420.2 [M+H] + , 1 H NMR (400MHz, DMSO) δ10.85(s,1H),8.53(s,1H),7.72(s,1H),7.56(d,J=5.1Hz,1H),7.48(d,J=7.9Hz,1H),7.28-7.20(m,1H),6.98(t,J=9 .9Hz,1H),6.63(d,J=12.2Hz,1H),3.90(s,3H),2.69(s,2H),1.99-1.88(m,4H),1.82-1.80(m,1H),1.75-1.61(m,2H),1.55-1.44(m,2H).

[0194] Example 18 Synthesis of Compound KH021

[0195]

[0196] Compound KH021: Compound KHB171 (204 mg, 0.80 mmol), KHB-234 (450 mg, 1.20 mmol), Pd(dppf)Cl2 (139 mg, 0.23 mmol), Cs2CO3 (1240 mg, 3.60 mmol) were dissolved in Dioxane (5 mL) and water (1 mL), and the reaction was stirred at 85°C for 3 hours under N2 protection. After the reaction was completed, the reaction system was diluted with 10 mL of water, extracted with ethyl acetate (10 mL x 3), the organic phases were combined, washed with saturated brine (10 mL), dried over anhydrous sodium sulfate, concentrated, and purified by C18 reverse phase column to obtain compound KH021. LCMS (ESI, m / z): 416.2 [M+H]+. 1H NMR (400MHz, DMSO) δ11.53(s,1H),8.85(s,1H),8.26(s,1H),7.98(d,J=10.8Hz,1H),7.82(d,J=8.0Hz,1H),7.74(d,J=8.0Hz, 1H), 7.58 (s, 1H), 7.36 (m, 2H), 7.01 (d, J = 7.6Hz, 1H), 6.87 (t, J = 7.2Hz, 1H), 6.80 (d, J = 7.6Hz, 1H), 4.26 (s, 3H), 1.96 (s, 3H).

[0197] Example 19 Synthesis of Compound KH022

[0198]

[0199] Compound KH022: Compound KHB238 (100.0 mg, 0.282 mmol), Pd(dppf)Cl2 (20.0 mg, 0.028 mmol) and potassium carbonate (117.0 mg, 0.846 mmol) were added to a mixed solution of compound KHB171 (76.0 mg, 0.282 mmol) in 1,4-dioxane (2.5 mL) and water (0.5 mL) in sequence, and stirred at 90°C for 3 h under nitrogen protection. After the reaction was complete, the reaction solvent was removed under reduced pressure to obtain a crude product, which was diluted with water, extracted with ethyl acetate, and the organic phase was washed twice with water and twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a crude product, which was then purified by reverse preparation to obtain compound KH022. LCMS (ESI, m / z): 416 [M+H]+. 1H NMR (400MHz, DMSO) δ11.57(s,1H),8.83(s,1H),7.89(s,1H),7.78(t,J=6.8Hz,2H),7.68(d,J=8.1Hz,1H),7.3 7-7.28(m,2H),6.98(d,J=7.4Hz,1H),6.92(t,J=7.5Hz,1H),6.86(t,J=11.1Hz,2H),4.25(s,3H),1.95(s,3H).

[0200] Example 20 BTK kinase inhibitory activity assay

[0201] 1. Cell culture and reagent testing

[0202] 1) Protein: human recombinant BTK.

[0203] 2) Reaction buffer: composed of 20 nM Tris, 50 nM NaCl, 10 mM MgCl2, 5 mM MnCl2, 1 mM DTT, 0.1 mM EDTA, 0.01% bovine serum albumin, 0.005% Tween-20 and 10% DMSO.

[0204] 3) Lance buffer: a commercially available reagent, its full English name is Lance assay buffer, and its brand is PerlinElmer.

[0205] 4) Instrument for determining the inhibitory activity of compounds on BTK kinase: Time-resolved Fluorescence Resonance Energy Transfer (TR-FRET).

[0206] 2. Test methods

[0207] 2.1 BTK kinase inhibitory activity assay

[0208] 1) Using a 96-well plate, 2 nM human recombinant BTK, small molecule inhibitors, 20 μM ATP and 1 μM peptide substrate (biotin-AVLESEEELYSSARQ-NH2) were placed in a reaction buffer (pH 7.4) and incubated at 23°C for 2 h.

[0209] 2) Add 25 μL of Lance buffer containing 1.2 equivalents of EDTA to quench the reaction.

[0210] 3) Add 62.5 nM streptavidin-APC (Streptavidin-XL665) and 0.67 nM Eu-labeled TK-antibody and incubate for 1 h.

[0211] 4) Use a multimode plate reader at an excitation wavelength of 330 nm (λ EX ) and 615nm, 665nm detection wavelength (λ EM ) to measure the TR-FRET signal.

[0212] 2.2 Data Analysis

[0213] Activity was determined by the fluorescence ratio at 665 nm and 615 nm. The kinase activity was measured at 11 different compound concentrations (3-fold dilution, concentration range between 1 μM and 0.017 nM) for each small molecule inhibitor. The result without adding small molecule inhibitor was used as negative control, and two enzyme-free controls were used to determine the baseline fluorescence level. The inhibition constant Ki (app) was obtained using the program BatchKi (Kuzmicet.al.Anal.Bi℃hem, 2000, 286:45-50). It was obtained according to the following formula

[0214] IC 50 :IC 50 = Ki(app) / (1+[ATP] / Km ATP )+[E] 总 / 2.

[0215] Km: Michaelis constant;

[0216] [E] Total: BTK concentration.

[0217] 2.3 Experimental Results

[0218] Table 1 Inhibitory activity of different compounds on BTK kinase

[0219]

[0220]

[0221] Example 21 BTKC481s kinase inhibitory activity assay

[0222] Compound BMS-986195 was prepared with reference to patent WO2016065226A1 (page 248 of the specification, Example 223).

[0223] 1. Cell culture and reagent testing

[0224] 1) Protein: Human recombinant BTKC481 S .

[0225] 2) Reaction buffer: composed of 20 nM Tris, 50 nM NaCl, 10 mM MgCl2, 5 mM MnCl2, 1 mM DTT, 0.1 mM EDTA, 0.01% bovine serum albumin, 0.005% Tween-20 and 10% DMSO.

[0226] 3) Lance buffer: a commercially available reagent, its full English name is Lance assay buffer, and its brand is PerlinElmer.

[0227] 4) Determination of the effect of compounds on BTK C481 S Instrument for inhibiting activity: Time-resolved Fluorescence Resonance Energy Transfer (TR-FRET).

[0228] 2. Test methods

[0229] 2.1 BTK C481S inhibitory activity assay

[0230] 1) Using a 384-well plate, 0.5 nM human recombinant BTK C481 S , small molecule inhibitors, 12 μM ATP and 1 μM biotin-labeled TK substrate were incubated in reaction buffer (pH 7.4) at 23°C for 90 min.

[0231] 2) Add 62.5 nM streptavidin-APC (Streptavidin-XL665) and 0.67 nM Eu-labeled TK-antibody and incubate at room temperature for 1 h.

[0232] 2.2 Data Analysis

[0233] Envision was used to detect the fluorescence signals at 615 nm and 665 nm. The activity was determined by the fluorescence ratio at 665 nm and 615 nm. The kinase activity was measured at 11 different compound concentrations (3-fold dilution, concentration range between 10 μM and 0.169 nM) for each small molecule inhibitor. The results without small molecule inhibitors were used as negative controls, and the baseline fluorescence level was determined using controls without kinase. The data were analyzed using 205 mode (4 Parameters Logistic Model) in IDBS's XLfit software to obtain IC 50 Value. IC 50 The formula is as follows:

[0234] fit=(A+((BA) / (1+((C / x)^D))))

[0235] res = (y-fit)

[0236] 4 Parameters

[0237] A:Bottom

[0238] B:Top

[0239] C:RelativeIC 50

[0240] D:HillSlope

[0241] 2.3 Experimental Results

[0242] Table 2 Inhibitory activity of different compounds on BTKC481s kinase

[0243] Compound <![CDATA[IC 50 (nM)]]> Compound <![CDATA[IC 50 (nM)]]> KH001 3.66 KH015 10.31 KH006 1.53 KH016 8.42 KH007 5.96 KH017 31.96 KH010 10.90 KH018 4.09 KH011 18.24 KH019 10.37 KH012 21.38 KH020 33.70 KH014 5.78 BMS-986195 658

Claims

1. A compound of formula I and its stereoisomers, solvates, pharmaceutically acceptable salts or prodrugs, characterized in that As shown in the structure of the compound of formula I: Wherein, A is selected from: R1a is selected from H, deuterium, halogen, amino, nitro, mercapto, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy; R1 is selected from H, deuterium, halogen, amino, nitro, sulfhydryl, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-8 cycloalkyl, C3-8 heterocyclyl, C5-10 aryl, C5-10 heteroaryl, wherein the C3-8 cycloalkyl, C3-8 heterocyclyl, C5-10 aryl, C5-10 heteroaryl are optionally substituted with deuterium, halogen, amino, nitro, sulfhydryl, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy; R2 is selected from H, deuterium, halogen, amino, nitro, mercapto, cyano, carboxyl, C1-6 alcohol, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, C1-6 hydroxyalkoxy, C3-8 cycloalkyl, C3-8 heterocyclyl, C5-10 aryl, C5-10 heteroaryl; R3 is selected from O, NH, S or (CH2)n, wherein n is an integer from 0 to 6; R4 is selected from a 5-10 membered monocyclic heterocyclic group or heteroaryl group, wherein the monocyclic heterocyclic group or heteroaryl group contains 2-4 heteroatoms selected from N, O, and S; optionally, the 5-10 membered heterocyclic group or heteroaryl group is substituted by H, halogen, amino, nitro, thiol, cyano, carboxyl, C1-6 alkyl, C1-6 haloalkyl, C1-6 deuterated alkyl, C1-6 hydroxyalkyl, C1-6 alkoxy, C1-6 haloalkoxy, or C1-6 hydroxyalkoxy.

2. The compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug of claim 1, characterized in that The R1a is H or C1-6 alkyl; more preferably H or methyl.

3. The compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug of claim 1, characterized in that The R1 is H, deuterium, halogen, amino, nitro, mercapto, cyano, carboxyl, C1-6 alkyl; more preferably, it is halogen, and the halogen is selected from F, Cl, Br, I; and most preferably, it is F.

4. The compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug of claim 1, characterized in that The R2 is H, C1-6 alcohol; preferably H, tert-butyl alcohol.

5. The compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug of claim 1, characterized in that The R3 is O, NH, (CH2)n, wherein n is 0.

6. The compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug of claim 1, characterized in that The R4 is 7. The compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug of claim 1, characterized in that As shown in the structure of the compound of formula II: wherein R1a is methyl or hydrogen, R1b is F or hydrogen, R1c is F or hydrogen, R2 is tert-butyl alcohol or hydrogen, R3 is -NH- or -O-, and R4 is 8. The compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug of claim 1, characterized in that Has the following structure:

9. A method for preparing compound III, comprising the following steps: Step (1): Alternatively, it further comprises step (2): Or the preparation method of the compound of formula III comprises the steps: Or further comprising the steps: Or the preparation method of the compound of formula III comprises the steps: The substituents are as defined above.

10. A pharmaceutical composition comprising a therapeutically effective dose of a compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug according to any one of claims 1 to 8 and one or more pharmaceutically acceptable carriers or excipients.

11. Use of the compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug according to any one of claims 1 to 8, or the pharmaceutical composition according to claim 9 in the preparation of a Bruton's tyrosinase inhibitor.

12. Use of the compound, stereoisomer, solvate, pharmaceutically acceptable salt or prodrug according to any one of claims 1 to 8, or the pharmaceutical composition according to claim 9 in the preparation of a drug for autoimmune disease, inflammation or tumor.

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