Compound as Cbl-b inhibitor and application thereof

Abstract

The invention belongs to the field of medical chemistry, relates to a compound serving as a Cbl-b inhibitor and application of the compound, and particularly provides a compound shown in a formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug of the compound. Processes for their preparation as well as pharmaceutical compositions containing these compounds and the use of these compounds or compositions for the treatment of Cbl-b mediated diseases. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of medicinal chemistry, and specifically relates to Cbl-b inhibitor compounds or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, methods for preparing the same, pharmaceutical compositions containing the same, and uses of the same or the compositions for treating Cbl-b-mediated diseases. Background Art

[0002] Casitas B lineage lymphoma proto-oncogene-b (Cbl-b) is an E3 ubiquitin ligase, mainly expressed in immune cells, and plays a key role in regulating effector T cell function. As an intracellular transduction factor, Cbl-b is located downstream of CD28 and CTLA4, and regulates innate and adaptive immunity through the action of E3 ubiquitin ligase (Chiang YJ et al., Nature, 2000, 403, 216-220.). CD28 is a star target in tumor immunotherapy, but direct targeting of CD28 is extremely challenging and may bring systemic immune risks. Drugs developed based on this target fail due to serious clinical adverse events. Cbl-b is an important regulatory factor downstream of CD28, and is also a major regulatory factor in T cell activation costimulatory and co-inhibitory pathways. Cbl-b not only ubiquitin-tags proteins for degradation and recycling in the proteasome, but also regulates the immune response by regulating the signaling pathways of effector cells (T cells and natural killer cells) (Magdalena Paolino et al., Nature, 2014, 507(7493), 508–512). Cbl-b plays an important role in T cell exhaustion. Studies have shown that Cbl-b is upregulated in exhausted CD8+ T cells, and Cbl-b deficiency enhances T cell activity and promotes tumor immunity (Jitendra Kumar et al. J Immunother Cancer, 2021, 9(1), e001688). Other studies have shown that Cbl-b deficiency enhances NK cell anti-tumor activity (Xuan Guo et al., J Immunother Cancer, 2021, 9(3), e001975). Inhibiting Cbl-b can relieve the immunosuppressive state of TME, promote T cell activation and recruitment by lowering the T cell activation threshold, and enhance the killing function of NK cells; promote the proliferation, differentiation and maturation of dendritic cells, and enhance the ability to present tumor antigens; improve the anti-tumor immune function of effector T cells, and reduce the influence of immunosuppressive cells such as Treg and MDSC on effector T cells, thereby strengthening the inflammatory response of the tumor microenvironment, mobilizing the entire immune system to attack tumor cells, and driving immune-mediated tumor growth inhibition (Augustin RC et al., J Immunother Cancer, 2023, 11(2), e006007).

[0003] Cancer immunotherapy has significantly improved the current status of tumor treatment, but there are still large limitations on the number of patients with indications, and drug resistance gradually develops with long-term use. Cbl-b in the CD28 signaling pathway is a key factor leading to immunosuppression in the tumor microenvironment. Targeting Cbl-b can increase immune cell activity and reverse drug resistance. There are currently some drug deployments targeting Cbl-b, such as siRNA and small molecule inhibitors. Preclinical studies have shown good immune activation and anti-tumor effects, but clinical translation still has certain challenges. Therefore, the development of new inhibitors targeting Cbl-b has positive significance for expanding tumor immunotherapy. Targeting Cbl-b may provide new immunotherapy methods for cancer patients by regulating the activation of immune cells (such as T cells), while expanding the treatment population for existing immune checkpoint therapies, and is a highly promising anti-tumor target. Summary of the Invention

[0004] One object of the present invention is to provide a class of compounds represented by general formula (I), general formula (Ia), and general formula (Ib) having Cbl-b inhibitory activity, or their isomers, pharmaceutically acceptable salts, solvates, crystals, or prodrugs.

[0005] Another object of the present invention is to provide a method for preparing the compounds of the present invention of general formula (I), general formula (Ia), general formula (Ib) or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs.

[0006] Another object of the present invention is to provide a composition comprising a compound of the present invention of general formula (I), general formula (Ia), general formula (Ib) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and a pharmaceutically acceptable carrier, as well as a composition comprising a compound of the present invention of general formula (I), general formula (Ia), general formula (Ib) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and one or more other drugs.

[0007] Another object of the present invention is to provide methods for treating Cbl-b-mediated diseases using the compounds of the present invention, or their isomers, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, and uses of the compounds of the present invention, or their isomers, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, in the preparation of medicaments for treating Cbl-b-mediated diseases.

[0008] In view of the above-mentioned invention objectives, the present invention provides the following technical solutions:

[0009] In a first aspect, the present invention provides a compound represented by general formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof,

[0010]

[0011] in,

[0012] X is selected from C or N;

[0013] Y is selected from C or N;

[0014] R 1 、R 2 、R 4 、R 5 each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, and dialkylamino;

[0015] Or two R 2 and the carbon atoms to which they are attached may together form a spirocycloalkyl group or a spiroheterocyclyl group, wherein the spirocycloalkyl group or the spiroheterocyclyl group is optionally substituted with one or more alkyl groups, alkoxy groups, haloalkoxy groups, haloalkyl groups, halogen groups, hydroxyl groups, carboxyl groups, cyano groups, amino groups, alkylacyl groups, aminoacyl groups, dialkylamino groups and oxo groups;

[0016] L is selected from a single bond, an alkylene group, an imino group, and an alkylene-imino group;

[0017] Ring A is selected from aryl, heteroaryl, cycloalkyl and heterocyclyl;

[0018] R 3 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, alkoxy, halogen, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkynyl, alkylsulfonyl, oxo, aminoacyl, alkylacylamino, cycloalkylaminoacyl, wherein the cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted with one or more hydrogen, alkyl, alkoxy, haloalkoxy, haloalkyl, halogen, hydroxy, carboxyl, cyano, amino, alkylacyl, aminoacyl, dialkylamino, and oxo groups;

[0019] is a single bond or a double bond, two of which All are single keys, or one is a single bond, and the other is a double bond; and

[0020] m, n and p are each independently selected from 0, 1, 2, and 3.

[0021] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof having the following structure of formula (Ia):

[0022]

[0023] Among them, L, ring A, R 3 、R 4 、R 5 and p are as defined in formula (I).

[0024] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof having the following structure of formula (Ib):

[0025]

[0026] in,

[0027] Ring B is selected from cycloalkyl, heterocyclyl, heteroaryl, wherein the cycloalkyl, heterocyclyl, heteroaryl is optionally substituted with one or more halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, and oxo groups;

[0028] Ring C is selected from aryl, heteroaryl, and the aryl, heteroaryl is optionally replaced by one or more R 3 Replacement; and

[0029] L, R 3 、R 4 、R 5 have the same definitions as described in general formula (I).

[0030] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: R 1 、R 2 、R 4 、R 5 Each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl and di-C 1-6 Alkylamino.

[0031] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: two R 2 and its connected carbon atoms can form spiro C 3-12 Cycloalkyl, spiro 3-12 membered heterocyclic group, the spiro C 3-12 Cycloalkyl, spiro 3-12 membered heterocyclic group are optionally substituted by one or more C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, halogenated C 1-6 Alkyl, fluorine, chlorine, bromine, iodine, hydroxyl, carboxyl, cyano, amino, C 1-6 Alkyl acyl, amino acyl, di-C 1-6 Alkylamino and oxo groups are substituted.

[0032] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: two R 2 and the carbon atom to which it is attached can together form a spirocyclopropyl group.

[0033] In some embodiments, the compound of the present invention is a compound of formula (I), formula (Ia), formula (Ib) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: L is selected from sub-C 1-6 Alkyl, imino, C 1-6 Alkyl-imino.

[0034] In some preferred embodiments, the compound of the present invention is a compound of general formula (I), general formula (Ia), general formula (Ib) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein: L is selected from -CH2-, -NH-, -NH-CH2-.

[0035] In some embodiments, the compound of the present invention is a compound of formula (I), formula (Ia) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: Ring A is selected from C 6-12 Aryl, 5-15 membered heteroaryl, C 3-12 cycloalkyl and 3-12 membered heterocyclic groups.

[0036] In some specific embodiments, the compound of the present invention is a compound of the general formula (I), the general formula (Ia) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: Ring A is selected from phenyl, naphthyl, anthracenyl, thiazolyl, pyridyl, benzopyranyl, benzo[d]thiazolyl, spiro[indoline-3,4'-piperidinyl]-2-one, 5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazinyl, 1,2,3,4-tetrahydroquinolinyl, 4,5,6 ,7-tetrahydro-1H-pyrrolo[3,2-c]pyridinyl, 2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazolyl, 1,2,3,4-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydroimidazo[1,2-a]pyrazinyl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazinyl, 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridinyl, cyclopropyl, piperidinyl, piperazinyl, 6-azaspiro[3.4]octanyl.

[0037] In some embodiments, the compound of the present invention is a compound of formula (Ib) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: Ring B is selected from C 3-12 Cycloalkyl, 3-12 membered heterocyclic group, 5-15 membered heteroaryl, the C 3-12 Cycloalkyl, 3-12 membered heterocyclyl, 5-15 membered heteroaryl, optionally substituted with one or more halogen, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino and oxo substitution;

[0038] Further preferably, ring B is selected from 5-10 membered heterocyclic groups, 5-6 membered heteroaryl groups, and the 5-10 membered heterocyclic groups, 5-6 membered heteroaryl groups are optionally substituted with one or more halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxy C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-3 Alkylamino, C 1-3Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino and oxo groups are substituted.

[0039] In some embodiments, the compound of the present invention is a compound of formula (Ib) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: Ring C is selected from C 6-12 Aryl, 5-15 membered heteroaryl, said C 6-12 Aryl, 5-15 membered heteroaryl are optionally substituted with one or more R 3 replace;

[0040] More preferably, ring C is selected from phenyl, 5-6 membered heteroaryl, and the phenyl, 5-6 membered heteroaryl is optionally replaced by one R 3 replace.

[0041] In some embodiments, the compound of the present invention is a compound of formula (I), formula (Ia), formula (Ib) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: R 3 Selected from hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, fluorine, chlorine, bromine, iodine, cyano, C 3-12 Cycloalkyl, 3-12 membered heterocyclic group, C 6-12 Aryl, 5-15 membered heteroaryl, C 3-12 Cycloalkyl C 2-6 Alkynyl, C 1-6 Alkylsulfonyl, oxo, aminoacyl, C 1-6 Alkyl acylamino, C 1-6 Cycloalkylaminoacyl, the C 3-12 Cycloalkyl, 3-12 heterocyclic, C 6-12 Aryl, 5-15 membered heteroaryl optionally substituted with one or more hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogen, hydroxyl, carboxyl, cyano, amino, C 1-6 Alkyl acyl, amino acyl, di-C 1-6 Alkylamino and oxo substitution;

[0042] More preferably, R 3 is selected from hydrogen, methyl, ethyl, vinyl, ethynyl, propynyl, methoxy, ethoxy, fluorine, chlorine, bromine, iodine, cyano, cyclopropyl, 5-10 membered heterocyclic radical, C 6-10aryl, 5-10 membered heteroaryl, cyclopropylethynyl, methylsulfonyl, oxo, aminoacyl, acetylamino, cyclopropylaminoacyl.

[0043] In some embodiments, the compound of the present invention is a compound of formula (I), formula (Ia) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: Selected from

[0044] The present invention provides the following specific compounds or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs:

[0045]

[0046]

[0047]

[0048] In a second aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof.

[0049] In some embodiments, the present invention provides a compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, and a pharmaceutical composition comprising the compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, for use in treating Cbl-b-mediated diseases.

[0050] In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof and a pharmaceutically acceptable carrier.

[0051] The compound of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug can be mixed with a pharmaceutically acceptable carrier, diluent or excipient to prepare a pharmaceutical preparation, so as to be suitable for oral or parenteral administration. Methods of administration include, but are not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal and oral routes. The preparation can be applied by any route, for example, by infusion or push injection, by an approach absorbed through the epithelium or mucocutaneous membranes (such as oral mucosa or rectum, etc.). Administration can be systemic or local. Examples of orally administered preparations include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The preparation can be prepared by methods known in the art and include carriers, diluents or excipients conventionally used in the field of pharmaceutical preparations.

[0052] In a third aspect, the present invention provides methods for treating Cbl-b-mediated diseases using compounds represented by formula (I), (Ia), or (Ib) of the present invention, or their isomers, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, or pharmaceutical compositions comprising the same, as well as their use in preparing medicaments for treating Cbl-b-mediated diseases.

[0053] In some preferred embodiments, the present invention provides methods for treating Cbl-b-mediated diseases and uses of compounds represented by Formula (I), (Ia), or (Ib), or their isomers, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, or pharmaceutical compositions comprising the same, in the preparation of medicaments for treating Cbl-b-mediated diseases, wherein the Cbl-b-mediated diseases include, but are not limited to, proliferative diseases, metabolic diseases, or blood diseases. In some embodiments, the Cbl-b-mediated disease described herein is cancer.

[0054] In some embodiments, the Cbl-b mediated diseases of the present invention include, but are not limited to, acoustic neuroma, adenocarcinoma, adrenal cancer, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendothelial sarcoma, angiosarcoma), adnexal cancer, benign monoclonal gammopathy, bile cancer (e.g., bile duct cancer), bladder cancer, breast cancer (e.g., breast adenocarcinoma, breast papillary carcinoma, breast cancer, medullary breast cancer, triple-negative breast cancer), brain cancer (e.g., meningioma; glioma, such as astrocytoma, oligodendroglioma; medulloblastoma), Bronchogenic carcinoma, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, chordoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial carcinoma, ependymoma, endothelial sarcoma (e.g., Kaposi's sarcoma, multiple idiopathic hemorrhagic sarcomas), endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarinoma), Ewing sarcoma sarcoma), eye cancer (e.g., intraocular melanoma, retinoblastoma), familial hypereosinophilia, gallbladder cancer, stomach cancer (e.g., gastric adenocarcinoma), gastrointestinal stromal tumor (GIST), head and neck cancer (e.g., head and neck squamous cell carcinoma), oral cancer (e.g., oral squamous cell carcinoma (OSCC), pharyngeal cancer (e.g., larynx cancer, pharynx cancer, nasopharyngeal cancer, oropharynx cancer)), hematopoietic cancer (e.g., leukemias such as acute lymphoblastic leukemia (ALL) (e.g., B-cell ALL, T-cell ALL), acute myeloid leukemia (AML) (e.g., B-cell AML, T-cell AML), chronic myeloid leukemia (CML) (e.g., B-cell CML, T-cell CML), and chronic lymphocytic leukemia (CLL) (e.g., B-cell CL) L, T-cell CLL); lymphomas such as Hodgkin lymphoma (HL) (e.g., B-cell HL, T-cell HL) and non-Hodgkin lymphoma (NHL) (e.g., B-cell NHL such as diffuse large cell lymphoma (DLCL) (e.g., diffuse large B-cell lymphoma (DLBCL)), follicular lymphoma, chronic lymphocytic leukemia / small lymphocytic lymphoma (CLL / SLL), mantle cell lymphoma (MCL), marginal zone B-cell lymphoma (e.g., mucosa-associated lymphoid tissue (MALT) lymphoma, nodular marginal zone B-cell lymphoma, splenic marginal zone B-cell lymphoma), primary mediastinal B-cell lymphoma, Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., "Woldenstrom's macroglobulinemia ( macroglobulinemia)”), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-lymphoblastic lymphoma, and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-lymphoblastic lymphoma / leukemia, peripheral T-cell lymphoma (PTCL) (e.g., cutaneous T-cell lymphoma (CTCL) (e.g., mycosis fungiodes, Sezary syndrome)), angioimmunoblastic T-cell lymphoma, extranodal natural killer T-cell lymphoma, enteropathy-type T-cell lymphoma, subcutaneous panniculitis-like T-cell lymphoma, anaplastic large cell lymphoma); mixtures of one or more leukemias / lymphomas as described above;and multiple myeloma (MM)), heavy chain diseases (e.g., alpha chain disease, gamma chain disease, mu chain disease), hemangioblastoma, inflammatory myofibroblastoma, immune cell amyloidosis, kidney cancer (e.g., Wilms' tumor, renal cell carcinoma), liver cancer (e.g., hepatocellular carcinoma (HCC), malignant hepatoma), lung cancer (e.g., bronchogenic carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma), leiomyosarcoma (LMS), mastocytosis (e.g., systemic mastocytosis), myelodysplastic syndrome (MDS), mesothelioma, myeloproliferative disorders (MPD) (e.g., polycythemia vera (PV), essential thrombocythemia (ET), idiopathic myeloid metaplasia (AMM), also known as Myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), hypereosinophilic leukemia syndrome (HES)), neuroblastoma, neurofibroma (e.g., multiple neurofibromatosis (NF) type 1 or type 2, schwannomatosis), neuroendocrine cancer (e.g., gastroenteropancreatic neuroendocrine tumor (GEP-NET), carcinoid tumor), osteosarcoma, ovarian cancer (e.g., cystadenocarcinoma, ovarian embryonal carcinoma, ovarian adenocarcinoma, ovarian clear cell carcinoma, ovarian serous cystadenocarcinoma), papillary adenocarcinoma, pancreatic cancer (e.g., pancreatic adenocarcinoma, intraductal papillary mucinous neoplasm (IPMN), islet cell tumor), penile cancer (e.g., Paget's disease of the penis and scrotum), disease), pinealoma, primary neuroectodermal tumor (PNT), prostate cancer (e.g., prostate adenocarcinoma), rectal cancer, rhabdomyosarcoma, salivary duct cancer, skin cancer (e.g., squamous cell carcinoma (SCC), keratoacanthoma (KA), melanoma, basal cell carcinoma (BCC)), small intestinal cancer (e.g., adnexal cancer), soft tissue sarcoma (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland cancer, sweat gland cancer, synovioma, testicular cancer (e.g., seminoma, testicular embryonal carcinoma), thyroid cancer (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid cancer), urethral cancer, vaginal cancer, and vulvar cancer (e.g., Paget's disease of the vulva), medulloblastoma, adenoid cystic carcinoma, melanoma, glioblastoma.

[0055] In some preferred embodiments, the present invention provides a method for treating a Cbl-b-mediated disease using a compound represented by Formula (I), Formula (Ia), or Formula (Ib) of the present invention, or an isomer, pharmaceutically acceptable salt, solvate, crystal, or prodrug thereof, or a pharmaceutical composition comprising the same, and a use thereof in preparing a medicament for treating a Cbl-b-mediated disease, wherein the Cbl-b-mediated disease includes, but is not limited to, breast cancer, esophageal cancer, bladder cancer, lung cancer, hematopoietic system cancer, lymphoma, medulloblastoma, medulloblastoma, rectal adenocarcinoma, colon cancer, gastric cancer, pancreatic cancer, liver cancer, adenoid cystic carcinoma, prostate cancer, lung cancer, head and neck squamous cell carcinoma, brain cancer, hepatocellular carcinoma, melanoma, oligodendroglioma, glioblastoma, testicular cancer, ovarian clear cell carcinoma, ovarian serous cystadenocarcinoma, thyroid cancer, multiple myeloma (AML), renal cell carcinoma, mantle cell lymphoma, triple-negative breast cancer, non-small cell lung cancer, hemoglobinopathy, diabetes, and obesity.

[0056] Definition of terms

[0057] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0058] The "hydrogen", "carbon" and "oxygen" in the compounds of the present invention include all isotopes thereof. Isotopes should be understood to include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include protium, tritium and deuterium, and isotopes of carbon include 12 C. 13 C and 14 C, oxygen isotopes include 16 O and 18 O etc.

[0059] "Isomers" in the present invention refer to molecules with identical atomic composition and bonding patterns but different three-dimensional arrangements, including but not limited to diastereomers, enantiomers, cis-trans isomers, and mixtures thereof, such as racemic mixtures. Many organic compounds exist in optically active forms, meaning they have the ability to rotate the plane of plane-polarized light. When describing optically active compounds, the prefixes D, L, R, or S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes D, L, or (+), (-) are used to designate the sign of rotation of plane-polarized light in the compound, with (-) or L indicating that the compound is levorotatory and the prefix (+) or D indicating that the compound is dextrorotatory. These stereoisomers have the same chemical structure but different stereostructures. Specific stereoisomers can be enantiomers, and a mixture of isomers is often referred to as an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, devoid of optical activity.

[0060] Depending on the choice of starting materials and process, the compounds of the present invention may exist as one of the possible isomers or as a mixture thereof, such as a racemate and a diastereomeric mixture (depending on the number of asymmetric carbon atoms). Optically active (R)- or (S)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

[0061] Any resulting mixture of stereoisomers can be separated into the pure or substantially pure geometric isomers, enantiomers, and diastereomers on the basis of the differences in the constituent physicochemical properties, for example, by chromatography and / or fractional crystallization.

[0062] The term "halogen" in the present invention refers to fluorine, chlorine, bromine, or iodine. The term "halo" in the present invention refers to substitution with fluorine, chlorine, bromine, or iodine.

[0063] The "alkyl" of the present invention refers to a straight-chain or branched saturated aliphatic hydrocarbon group, preferably a straight-chain or branched group containing 1 to 6 carbon atoms, more preferably a straight-chain or branched group containing 1 to 3 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, etc. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be at any available point of attachment.

[0064] In the present invention, "carbonyl" and "acyl" both refer to -C(O)-.

[0065] The "sulfonyl group" of the present invention refers to -S(O)2-.

[0066] The "sulfonamide group" of the present invention refers to -S(O)2NH-.

[0067] The "haloalkyl group" of the present invention refers to an alkyl group substituted with at least one halogen.

[0068] The "hydroxyalkyl group" of the present invention refers to an alkyl group substituted with at least one hydroxy group.

[0069] "Alkoxy" herein refers to -O-alkyl. Non-limiting examples of alkoxy groups include methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy, and the like. Alkoxy groups may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

[0070] The "cycloalkyl" of the present invention refers to a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups can be substituted or unsubstituted monocyclic rings having 3 to 12 carbon atoms or fused, bridged or spiro bicyclic or tricyclic saturated hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0071] "Heterocyclyl" of the present invention refers to a group ("3-12 membered heterocyclyl") of a 3- to 12-membered non-aromatic ring system having 1 to 4 ring heteroatoms (wherein each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon). In heterocyclyl groups comprising one or more nitrogen atoms, the point of attachment may be a carbon or nitrogen atom, as long as valence permits. The heterocyclyl group may be monocyclic ("monocyclic heterocyclyl") or a fused, bridged or spiral ring system (e.g., a bicyclic system (also known as a "bicyclic heterocyclyl")), and may be saturated or may be partially unsaturated. Suitable heterocyclyl groups include, but are not limited to, piperidinyl, azetidinyl, aziridine, tetrahydropyrrolyl, piperazinyl, dihydroquinazolinyl, oxacyclopropyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, 6-azaspiro [3.4] octanyl, Each example of heterocyclyl may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

[0072] The term "aryl" herein refers to an aromatic system that may comprise a monocyclic or fused polycyclic ring, preferably a monocyclic or fused bicyclic ring, containing 6 to 12 carbon atoms, preferably about 6 to about 10 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, fluorenyl, and indanyl. Aryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any available point of attachment.

[0073] "Heteroaryl" herein refers to an aryl group in which at least one carbon atom is replaced by a heteroatom. The heteroaryl group may have a monocyclic ring or a fused, bridged, or spiro polycyclic ring system, in which at least one ring is aromatic. Preferably, the heteroaryl group is composed of 5-15 atoms (5-15 membered heteroaryl), and more preferably, 5-10 atoms (5-10 membered heteroaryl), wherein the heteroatom is O, S, or N. The heteroaryl groups include, but are not limited to, imidazolyl, pyrrolyl, furyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, indolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, benzopyrazolyl, benzimidazolyl, benzofuranyl, benzopyranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, quinolyl, isoquinolyl, quinazolinyl, cinnolinyl, quinoxalinyl, benzoxazinyl, benzothiazinyl, imidazopyridinyl, pyrimidopyrazolyl, pyrimidoimidazolyl, benzo[d]thiazolyl, spiro[indole] [1,2,4]triazolo[1,5-a]pyrazinyl, 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridinyl, 2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazolyl, 1,2,3,4-tetrahydroisoquinolinyl, 5,6,7,8-tetrahydroimidazo[1,2-a]pyrazinyl, 5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazinyl, 4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridinyl, etc. Heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any available point of attachment.

[0074] The "pharmaceutically acceptable salts" of the present invention 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.

[0075] In the present invention, "solvate" refers to a complex formed by the combination of a solute (e.g., an active compound, a salt of an active compound) and a solvent (e.g., water). The solvent is any solvent known or readily identified by one skilled in the art. If the solvate is water, the solvate is typically referred to as a hydrate, such as a hemihydrate, monohydrate, dihydrate, trihydrate, or alternatives thereof.

[0076] The in vivo effects of the compounds of formula (I), (Ia), (Ib) may be exerted in part by one or more metabolites formed in the human or animal body after administration of the compounds of formula (I), (Ia), (Ib). As described above, the in vivo effects of the compounds of formula (I), (Ia), (Ib) may also be exerted via metabolism of precursor compounds ("prodrugs"). "Prodrugs" of the present invention refer to compounds that are converted into compounds of the present invention under physiological conditions in an organism due to reaction with enzymes, gastric acid, etc., i.e., compounds that are converted into compounds of the present invention by oxidation, reduction, hydrolysis, etc. of an enzyme and / or compounds that are converted into compounds of the present invention by hydrolysis reactions, etc., such as gastric acid.

[0077] The term "crystalline" in the present invention refers to a solid whose internal structure is formed by regularly repeating constituent atoms (or groups thereof) in three dimensions, as distinguished from an amorphous solid that does not have such a regular internal structure.

[0078] A "pharmaceutical composition" of the present invention refers to a mixture comprising any of the compounds described herein, including corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts, or chemically protected forms thereof, and one or more pharmaceutically acceptable carriers and / or one or more additional drugs. The purpose of a pharmaceutical composition is to facilitate administration of the compound to an organism. Such compositions are typically used to prepare medicaments for treating and / or preventing diseases mediated by one or more kinases.

[0079] The term "pharmaceutically acceptable carrier" as used herein refers to a carrier that does not significantly irritate an organism and does not interfere with the biological activity and properties of the administered compound, and includes all solvents, diluents or other excipients, dispersants, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, and the like. This excludes any conventional carrier medium that is incompatible with the compound of the invention. Examples of pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, cellulose, and cellulose acetate; malt, gelatin, and the like.

[0080] The "excipient" of the present invention refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols. DETAILED DESCRIPTION

[0081] The present invention will be further described in detail below with reference to the following examples, but the present invention is not limited to these examples. Unless otherwise specified, the materials used in the following examples were all commercially available.

[0082] Intermediate 1: 3-(1-(3-bromophenyl)-3-methylcyclobutyl)-4-methyl-4H-1,2,4-triazole

[0083]

[0084] Step 1: Preparation of methyl 1-(3-bromophenyl)-3-methylcyclobutane-1-carboxylate

[0085]

[0086] Methyl 2-(3-bromophenyl)acetate (9.2 g, 40.2 mmol, 1.0 eq) was dissolved in anhydrous N,N-dimethylformamide (150 mL). 1,3-Dibromo-2-methylpropane (8.6 g, 40.2 mmol, 1.0 eq) was added. Sodium hydride (1.93 g, 80.4 mmol, 2.0 eq) was added portionwise at 0°C and stirred at room temperature for 2 h. The reaction was monitored by LC-MS. After completion, the mixture was quenched with saturated aqueous ammonium chloride and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 7.85 g of the title compound. ESI-MS m / z: 282.9 [M+H] + .

[0087] Step 2: Preparation of 1-(3-bromophenyl)-3-methylcyclobutane-1-carboxylic acid hydrazide

[0088]

[0089] Dissolve methyl 1-(3-bromophenyl)-3-methylcyclobutyl-1-carboxylate (7.85 g, 27.84 mmol, 1.0 eq) in ethanol (110 mL), add hydrazine hydrate (28.7 mL), and stir overnight at 80°C under argon. Monitor the reaction using LC-MS. After completion, concentrate the reaction mixture to yield 7.85 g of the title compound. ESI-MS m / z: 283.1 [M+H] + .

[0090] Step 3: Preparation of 2-(1-(3-bromophenyl)-3-methylcyclobutane-1-carbonyl)-N-methylhydrazine-1-thiocarboxamide

[0091]

[0092] Dissolve 1-(3-bromophenyl)-3-methylcyclobutyl-1-carboxylic acid hydrazide (7.85 g, 27.84 mmol, 1.0 eq) in anhydrous tetrahydrofuran (205 mL), add methyl isothiocyanate (6.10 g, 83.52 mmol, 3.0 eq), and stir at 80°C for 2 hours. Monitor the reaction by LC-MS. After completion, quench the reaction with water and extract with ethyl acetate. The combined organic phases are washed three times with water, dried over anhydrous sodium sulfate, and concentrated to yield 12.22 g of the title compound. ESI-MS m / z: 356.2 [M+H] + .

[0093] Step 4: Preparation of 5-(1-(3-bromophenyl)-3-methylcyclobutyl)-4-methyl-4H-1,2,4-triazole-3-thiol

[0094]

[0095] 2-(1-(3-bromophenyl)-3-methylcyclobutyl-1-carbonyl)-N-methylhydrazinyl-1-thiocarboxamide (12.22 g, 34.42 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (73 mL). Aqueous sodium hydroxide (279 mL, 275.36 mmol, 8.0 eq) was added and stirred at room temperature for 16 h. The reaction was monitored by LC-MS. After completion, the pH was adjusted to 3 with dilute hydrochloric acid, and ethyl acetate was added for extraction. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, and concentrated to yield 11.32 g of the title compound. ESI-MS m / z: 338.1 [M+H] + .

[0096] Step 5: Preparation of 3-(1-(3-bromophenyl)-3-methylcyclobutyl)-4-methyl-4H-1,2,4-triazole

[0097]

[0098] 5-(1-(3-bromophenyl)-3-methylcyclobutyl)-4-methyl-4H-1,2,4-triazole-3-thiol (11.32 g, 33.45 mmol, 1.0 eq) was dissolved in anhydrous dichloromethane (205 mL). A solution of hydrogen peroxide (5.69 g, 167.26 mmol, 5.0 eq) in acetic acid (41 mL) was added dropwise at 0°C and stirred at room temperature for 16 h. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water, the pH was adjusted to 10 with sodium hydroxide solution, and extraction was performed with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 5.6 g of the title compound. ESI-MS m / z: 306.1 [M+H] + .

[0099] Intermediate 2: 3-((1S,3S)-1-(3-bromophenyl)-3-methylcyclobutyl)-4-methyl-4H-1,2,4-triazole

[0100]

[0101] The racemate of 3-(1-(3-bromophenyl)-3-methylcyclobutyl)-4-methyl-4H-1,2,4-triazole (200 mg) was separated using a chiral preparative column. (Method: Chiral column: Chiralpak IK-3 50x4.6 mm ID, 3 μm, elution: isocratic: B 40%, A = carbon dioxide, B = methanol (0.05% diethanolamine)) to afford 93 mg of the title compound. ESI-MS m / z: 306.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ8.30(s,1H),7.50-7.44(m,2H),7.38-7.32(m,2H) ,3.17(s,3H),2.86-2.74(m,2H),2.55-2.52(m,3H),1.06(d,J=4.0Hz,3H).

[0102] Intermediate 3: Methyl 2-(bromomethyl)-5-formyl-3-(trifluoromethyl)benzoate

[0103]

[0104] Step 1: Preparation of methyl 5-iodo-2-methyl-3-(trifluoromethyl)benzoate

[0105]

[0106] Methyl 2-methyl-3-(trifluoromethyl)benzoate (25 g, 114.6 mmol, 1.0 eq) was dissolved in trifluoromethanesulfonic acid (210 mL). N-iodosuccinimide (33.5 g, 148.96 mmol, 1.3 eq) was added at 10°C and stirred at 60°C for 3 h. The reaction was monitored by LC-MS. After completion, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 14.7 g of the title compound. ESI-MS m / z: 346.1 [M+H]. + .

[0107] Step 2: Preparation of 3-(methoxycarbonyl)-4-methyl-5-(trifluoromethyl)benzoic acid

[0108]

[0109] Methyl 5-iodo-2-methyl-3-(trifluoromethyl)benzoate (14.5 g, 42 mmol, 1.0 eq) was dissolved in anhydrous N,N-dimethylformamide (140 mL). Oxalic acid (5.6 g, 63 mmol, 1.5 eq), acetic anhydride (6.4 g, 63 mmol, 1.5 eq), N,N-diisopropylethylamine (8.1 g, 63 mmol, 1.5 eq), palladium acetate (0.94 g, 4.2 mmol, 0.1 eq), and 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (1.2 g, 2.1 mmol, 0.05 eq) were added sequentially. The mixture was stirred at 100°C overnight under argon. The reaction was monitored by LC-MS. After completion, the reaction was filtered, the filtrate was concentrated, and purified by reverse-phase column chromatography to obtain 5.6 g of the title compound. ESI-MS m / z: 261.2 [MH] - .

[0110] Step 3: Preparation of 4-(bromomethyl)-3-(methoxycarbonyl)-5-(trifluoromethyl)benzoic acid

[0111]

[0112] Dissolve 3-(Methoxycarbonyl)-4-methyl-5-(trifluoromethyl)benzoic acid (5.5 g, 21 mmol, 1.0 eq) in carbon tetrachloride (110 mL). Add N-bromosuccinimide (5.5 g, 31.5 mmol, 1.5 eq) and dibenzoyl peroxide (1.5 g, 6.3 mmol, 0.3 eq) sequentially. Stir at 80°C for 3 h. Monitor the reaction by LC-MS. After completion, concentrate the product and purify by column chromatography to obtain 6.0 g of the title compound. ESI-MS m / z: 339.1 [MH] - .

[0113] Step 4: Preparation of methyl 2-(bromomethyl)-5-(hydroxymethyl)-3-(trifluoromethyl)benzoate

[0114]

[0115] Dissolve methyl 2-(bromomethyl)-5-(hydroxymethyl)-3-(trifluoromethyl)benzoate (6.0 g, 17.6 mmol, 1.0 eq) in anhydrous tetrahydrofuran (80 mL). Add borane tetrahydrofuran solution (44 mL, 44 mmol, 2.5 eq) in an ice bath and stir overnight at room temperature. Monitor the reaction by LC-MS. After completion, quench with methanol, concentrate, and purify by column chromatography to obtain 4.4 g of the title compound. ESI-MS m / z: 326.9 [M+H] + .

[0116] Step 5: Preparation of methyl 2-(bromomethyl)-5-formyl-3-(trifluoromethyl)benzoate

[0117]

[0118] Methyl 2-(bromomethyl)-5-(hydroxymethyl)-3-(trifluoromethyl)benzoate (4.4 g, 13.5 mmol, 1.0 eq) was dissolved in dichloromethane (100 mL), and Dess-Martin reagent (8.5 g, 20.2 mmol, 1.5 eq) was added. The mixture was stirred at room temperature for 3 h. LC-MS was used to monitor the reaction. After completion, the mixture was diluted with dichloromethane and extracted with saturated sodium thiosulfate. The organic phases were combined, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 3.8 g of the title compound. ESI-MS m / z: 325.1 [M+H] + .

[0119] Example 1: 6-((Benzo[d]thiazol-2-ylamino)methyl)-2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one

[0120]

[0121] Step 1: Preparation of 6-((Benzo[d]thiazol-2-ylamino)methyl)-4-(trifluoromethyl)isoindol-1-one

[0122]

[0123] Benzo[d]thiazol-2-amine (0.23 g, 1.54 mmol, 1.0 eq) was dissolved in dichloromethane (10 mL). Triethylamine (0.94 g, 9.24 mmol, 6.0 eq) and sodium triacetoxyborohydride (1.95 g, 9.24 mmol, 6.0 eq) were added sequentially. After stirring at room temperature for 10 min, a solution of intermediate 3 (0.5 g, 1.54 mmol, 1.0 eq) in dichloromethane (2 mL) was added at 0°C and stirred overnight at room temperature. The reaction was monitored by LC-MS. After completion, the mixture was quenched with saturated ammonium chloride solution under an ice bath. The mixture was extracted with ethyl acetate, and the combined organic phases were washed three times with water, dried over anhydrous sodium sulfate, and concentrated. Methanol (10 mL) was added for dissolution. Ammonia in methanol (3 mL) was added under an ice bath and stirred at room temperature for 4 h. The reaction was monitored by LC-MS. After completion, the mixture was concentrated directly and purified by column chromatography to afford 0.15 g of the title compound. ESI-MS m / z:364.1[M+H] + .

[0124] Step 2: Preparation of 6-((Benzo[d]thiazol-2-ylamino)methyl)-2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one

[0125]

[0126] 6-((Benzo[d]thiazol-2-ylamino)methyl)-4-(trifluoromethyl)isoindol-1-one (133 mg, 0.37 mmol, 1.0 eq) was dissolved in dioxane (6 mL). Intermediate 1 (115 mg, 0.37 mmol, 1.0 eq), palladium acetate (10 mg, 0.037 mmol, 0.1 eq), 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene (42.3 mg, 0.073 mmol, 0.2 eq), and cesium carbonate (357.7 mg, 1.098 mmol, 3.0 eq) were added sequentially. The mixture was stirred at 120°C under argon overnight. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to afford 36 mg of the title compound. ESI-MS m / z:589.2[M+H] + . 1 H NMR (400MHz, DMSO-d6): δ8.72-8.66(m,1H),8.33-8.25(m,1H),8.05(s,2H),7.95-7.88(m,1H),7.76 -7.67(m,2H),7.45-7.35(m,2H),7.24-7.21(m,1H),7.11-7.02(m,2H),5.24-5.11(m,2H),4.81(d,J= 8.0Hz,2H),4.63(d,J=16.0Hz,1H),4.48-4.44(m,1H),3.50-3.40(m,1H),3.22-3.19(m,1H),2.89-2 .82(m,1H),2.70-2.64(m,1H),2.35-2.28(m,2H),1.71(s,1H),1.28-1.21(m,1H),1.13-1.04(m,1H).

[0127] Example 2: 6-((5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one

[0128]

[0129] The preparation method was the same as that of Example 1, except that the raw material benzo[d]thiazol-2-amine was replaced with 5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine to obtain 19 mg of the title compound. ESI-MS m / z: 563.0 [M+H] + . 1 HNMR (400MHz, DMSO-d6): δ8.29(s,1H),8.11-7.98(m,3H),7.91(s,1H),7.69(d,J=20.0Hz,1H),7.44(d,J=16.0Hz,1H),7.13(d,J=8.0Hz, 1H),5.22(s,2H),4.14(s,2H),3.99(s,2H),3.78(s,2H),3.22(s,3H),3.02(s,2H),2.86(s,2H),2.61-2.54(m,3H),1.09(d,J=4.0Hz,3H).

[0130] Example 3: 1'-((2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindolin-5-yl)methyl)spiro[indoline-3,4'-piperidin]-2-one

[0131]

[0132] Step 1: Preparation of 3-oxo-7-(trifluoromethyl)isoindoline-5-carbaldehyde

[0133]

[0134] Intermediate 3 (0.72 g, 2.2 mmol, 1.0 eq) was dissolved in methanol (10 mL). Ammonia-methanol solution (3.8 mL, 7.6 mmol, 3.5 eq) was added under ice-cooling conditions and stirred at room temperature overnight. The reaction was monitored by LC-MS. After completion, the product was concentrated and purified by column chromatography to obtain 0.6 g of the title compound. ESI-MS m / z: 229.9 [M+H] + .

[0135] Step 2: Preparation of 2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindoline-5-carbaldehyde

[0136]

[0137] 3-Oxo-7-(trifluoromethyl)isoindoline-5-carbaldehyde (0.35 g, 1.15 mmol, 1.2 eq) was dissolved in dioxane (12 mL), and intermediate 1 (0.22 g, 0.96 mmol, 1.0 eq), methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl) palladium (II) (0.16 g, 0.19 mmol, 0.2 eq), 2-dicyclohexylphosphino-2,6-diisopropoxy-1,1-biphenyl (0.18 g, 0.38 mmol, 0.4 eq) and cesium carbonate (0.63 g, 1.92 mmol, 2.0 eq) were added in sequence, and stirred at 100 ° C for 2 h under argon protection in a microwave. The reaction was monitored by LC-MS. After completion, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 32 mg of the title compound. ESI-MS m / z: 455.1 [M+H] + .

[0138] Step 3: Preparation of 1'-((2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindolin-5-yl)methyl)spiro[indoline-3,4'-piperidin]-2-one

[0139]

[0140] Spiro[indoline-3,4'-piperidin]-2-one (22.3 mg, 0.049 mmol, 1.0 eq) was dissolved in dichloromethane (3 mL). Triethylamine (29.8 mg, 0.29 mmol, 6.0 eq) and sodium triacetoxyborohydride (82.9 mg, 0.39 mmol, 8.0 eq) were added sequentially. After stirring at room temperature for 10 min, a dichloromethane solution (1 mL) of 2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindoline-5-carbaldehyde (9.9 mg, 0.049 mmol, 1.0 eq) was added at 0°C and stirred at room temperature overnight. The reaction was monitored by LC-MS. After completion, the reaction was quenched with saturated ammonium chloride solution in an ice bath, extracted with ethyl acetate, and the combined organic phases were washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to obtain 3 mg of the title compound. ESI-MS m / z: 641.2 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ10.36(s,1H),8.32-8.27(m,1H),8.09-8.04(m,1H),8.00(s,1H),7.70-7 .65(m,1H),7.54-7.34(m,3H),7.23-7.09(m,2H),6.98-6.94(m,1H),6.85(d,J=8.0Hz,1H),5.21- 5.17(m,2H),4.65(d,J=16.0Hz,1H),3.85(s,2H),3.49(s,2H),3.23-3.18(m,2H),2.94-2.82(m,4 H),2.70-2.62(m,2H),1.87-1.77(m,2H),1.75-1.64(m,3H),1.28-1.21(m,1H),1.14-1.05(m,2H).

[0141] Example 4: 2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-(3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindolin-1-one

[0142]

[0143] Step 1: Preparation of 6-(chloromethyl)-4-(trifluoromethyl)isoindolin-1-one

[0144]

[0145] Dissolve 6-hydroxymethyl-4-(trifluoromethyl)isoindolin-1-one (0.81 g, 3.5 mmol, 1.0 eq) in 1,2-dichloroethane (20 mL). Under argon, slowly add thionyl chloride (0.8 μL, 11 mmol, 3.0 eq) dropwise. Stir at 50°C for 3.5 h. Monitor the reaction by LC-MS. After completion, concentrate the product and purify by column chromatography to obtain 0.87 g of the title compound. ESI-MS m / z: 250.0 [M+H] + .

[0146] Step 2: Preparation of 6-((3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0147]

[0148] 6-(Chloromethyl)-4-(trifluoromethyl)isoindolin-1-one (0.33 g, 1.31 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (6 mL). N,N-diisopropylethylamine (0.5 g, 3.87 mmol, 3.0 eq) and 3-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine (0.19 g, 1.28 mmol, 1.1 eq) were added sequentially and stirred at 50°C for 9 h. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 0.42 g of the title compound. ESI-MS m / z: 352.1 [M+H]. + .

[0149] Step 3: Preparation of 2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-(3-methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindolin-1-one

[0150]

[0151] 6-((3-Methyl-5,6-dihydro-[1,2,4]triazolo[4,3-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindol-1-one (0.16 g, 0.44 mmol, 1.0 eq) was dissolved in dioxane (8 mL), and intermediate 1 (0.14 g, 0.47 mmol, 1.1 eq), methanesulfonic acid (2-dicyclohexylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (82 mg, 0.09 mmol, 0.2 eq) and cesium carbonate (0.47 g, 1.44 mmol, 3.3 eq) were added in sequence, and the mixture was stirred at 100°C under argon protection overnight. The reaction was monitored by LC-MS. After completion, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to obtain 18 mg of the title compound. ESI-MS m / z: 577.2 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ8.29(s,1H),8.10-7.96(m,3H),7.69(d,J=8.0Hz,1H),7.48-7.40(m,1H),7.13(d,J=8.0Hz,1H),5.22(s,2H), 3.96(s,2H),3.9-3.87(m,2H),3.73(s,2H),3.22(s,3H),2.95-2.80(m,4H),2.56(d,J=8.0Hz,3H),2.29(s,3H),1.09(d,J=4.0Hz,3H).

[0152] Example 5: 2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-(3-methyl-3,4-dihydroquinolin-1(2H)-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0153]

[0154] The preparation method was the same as that of Example 4, except that the raw material 3-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine was replaced with 3-methyl-1,2,3,4-tetrahydroquinoline to obtain 13 mg of the title compound. ESI-MS m / z: 588.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ8.28(s,1H),8.08(s,1H),7.92(s,1H),7.84(s,1H),7.64(d,J=8.0Hz,1H),7.47-7 .39(m,1H),7.10(d,J=8.0Hz,1H),6.94(d,J=8.0Hz,1H),6.91-6.85(m,1H),6.54-6.48(m,1H),6.45(d,J=8 .0Hz,1H),5.19(s,2H),4.70(d,J=12.0Hz,2H),3.48(s,1H),3.41-3.38(m,1H),3.20(s,3H),3.12-3.04(m, 2H),2.89-2.74(m,2H),2.57-2.57(m,3H),2.16-2.05(m,1H),1.08(d,J=4.0Hz,3H),1.04(d,J=8.0Hz,3H).

[0155] Example 6: 2-(3-(3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-((1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0156]

[0157] The preparation method was the same as that of Example 4, except that the raw material 3-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine was replaced with 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine to obtain 20 mg of the title compound. ESI-MS m / z: 561.3 [M+H] + . 1 HNMR (400MHz, DMSO-d6): δ10.36 (s, 1H), 8.30 (d, J = 12.0Hz, 1H), 8.05-7.96 (m, 3H), 7.77-7 .68(m,1H),7.49-7.31(m,1H),7.19-7.04(m,1H),6.51(s,1H),5.70(s,1H),5.20-5.17(m,2 H),4.70-4.43(m,2H),3.85(s,2H),3.49(s,2H),3.39-3.37(m,2H),3.22(s,1H),3.03-2.93 (m,1H),2.92-2.80(m,1H),2.78-2.69(m,2H),2.64-2.55(m,3H),1.71(s,2H),1.09(s,1H).

[0158] Example 7: 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-((1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0159]

[0160] Step 1: Preparation of 6-(hydroxymethyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one

[0161]

[0162] 6-(Hydroxymethyl)-4-(trifluoromethyl)isoindolin-1-one (0.50 g, 2.16 mmol, 1.0 eq) was dissolved in dioxane (20 mL), and intermediate 2 (0.58 g, 1.88 mmol, 0.87 eq), methanesulfonic acid (2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl)(2-amino-1,1'-biphenyl-2-yl) palladium (II) (0.27 g, 0.32 mmol, 0.15 eq), 2-dicyclohexylphosphino-2',6'-diisopropoxy-1,1'-biphenyl (0.29 g, 0.63 mmol, 0.29 eq) and cesium carbonate (1.27 g, 3.89 mmol, 1.80 eq) were added in sequence, and the mixture was stirred at 100 ° C under argon protection overnight. The reaction was monitored by LC-MS. After completion, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 0.54 g of the title compound. ESI-MS m / z: 457.2 [M+H] + .

[0163] Step 2: Preparation of 6-(chloromethyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one

[0164]

[0165] Dissolve 6-(hydroxymethyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one (0.54 g, 1.18 mmol, 1.0 eq) in 1,2-dichloroethane (10 mL). Under argon protection, slowly add thionyl chloride (0.69 mg, 5.80 mmol, 4.9 eq) dropwise. Stir at 50°C for 3.5 h. Monitor the reaction by LC-MS. After completion, concentrate the product and purify by column chromatography to obtain 0.58 g of the title compound. ESI-MS m / z: 475.1 [M+H]. + .

[0166] Step 3: Preparation of 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-((1,4,6,7-tetrahydro-5H-pyrrolo[3,2-c]pyridin-5-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0167]

[0168] 6-(Chloromethyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one (0.11 g, 0.23 mmol, 1.0 eq) was dissolved in anhydrous acetonitrile (6 mL). 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine (32 mg, 0.26 mmol, 1.1 eq) and N,N-diisopropylethylamine (0.18 g, 1.39 mmol, 6.0 eq) were added sequentially. The mixture was stirred at 40°C under argon overnight. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to obtain 47 mg of the title compound. ESI-MS m / z:561.2[M+H] + . 1 H NMR (400MHz, DMSO-d6): δ10.37(s,1H),8.34-8.26(m,1H),8.13-7.94(m,3H),7.75-7 .65(m,1H),7.51-7.39(m,1H),7.19-7.07(m,1H),6.59-6.46(m,1H),5.78-5.64(m,1H ),5.31-5.15(m,2H),3.95-3.79(m,2H),3.41-3.33(m,3H),3.31(s,1H),3.25-3.19(m ,2H),2.96-2.80(m,2H),2.79-2.69(m,2H),2.63-2.55(m,4H),1.09(d,J=4.0Hz,3H).

[0169] Example 8: 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-(2-methyl-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0170]

[0171] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 2-methyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine to obtain 120 mg of the title compound. ESI-MS m / z: 577.3 [M+H] + . 1HNMR (500MHz, DMSO-d6): δ8.28(s,1H),8.06(s,2H),8.01(s,1H),7.69(d,J =10.0Hz,1H),7.44(d,J=10.0Hz,1H),7.13(m,1H),5.21(s,1H),4.05(s,2H) ,3.71(s,2H),3.22(s,4H),2.98(d,J=5.0Hz,3H),2.86(d,J=5.0Hz,3H),2. 66-2.62(m,1H),2.56(d,J=10.0Hz,4H),2.20(s,1H),1.09(d,J=5.0Hz,3H).

[0172] Example 9: 6-((2-bromo-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0173]

[0174] Step 1: Preparation of 2-bromo-[1,2,4]triazolo[1,5-a]pyrazine

[0175]

[0176] [1,2,4]triazolo[1,5-A]pyrazin-2-amine (5.40 g, 40 mmol, 1.0 eq) was dissolved in a mixture of acetic acid (70 mL) and water (35 mL). Stirring was continued at 0°C for 10 minutes, followed by the slow dropwise addition of hydrogen bromide (35 mL). Stirring was continued for 1.5 hours, followed by the slow dropwise addition of sodium nitrite solution (2.04 g, 45.2 mmol, 1.1 eq). After stirring for 2 hours, cuprous bromide (1.44 g, 10 mmol, 0.25 eq) was added. The temperature was slowly raised to 80°C, and heated with stirring for 4 hours. The reaction was monitored by LC-MS. After completion, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 0.28 g of the title compound. ESI-MS m / z: 199.0 [M+H]. + Step 2: Preparation of 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine

[0177]

[0178] Dissolve 2-bromo-[1,2,4]triazolo[1,5-a]pyrazine (0.25 g, 1.3 mmol, 1.0 eq) in methanol (10 mL). Slowly add a solution of lithium borohydride in tetrahydrofuran (1.25 mL, 2.5 mmol, 2.0 eq) dropwise and stir at room temperature for 3.5 h. LC-MS monitoring of the reaction was performed. After completion, the mixture was quenched with water, dried to dryness, and extracted with ethyl acetate. The combined organic phases were washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 0.17 g of the title compound. ESI-MS m / z: 203.0 [M+H] + .

[0179] Step 3: Preparation of 6-((2-bromo-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0180]

[0181] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine to obtain 35 mg of the title compound. ESI-MS m / z: 641.1 [M+H] + . 1 H NMR(500MHz,DMSO-d6):8.29(s,1H),8.06(d,J=10.0Hz,2H),8.01(s,1H), 7.69(d,J=10.0Hz,1H),7.46-7.42(m,1H),7.13(d,J=10.0Hz,1H),5.21(s, 2H),4.16-4.10(m,2H),3.99(s,2H),3.80(s,2H),3.20(s,3H),3.04-2.96( m,2H),2.87(d,J=5.0Hz,2H),2.56(d,J=5.0Hz,3H),1.09(d,J=5.0Hz,3H).

[0182] Example 10: 7-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-5,6,7-8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carbonitrile

[0183]

[0184] The compound from Example 9 (65 mg, 0.1 mmol, 1.0 eq) was dissolved in N,N-dimethylacetamide (3 mL). Zinc cyanide (14.3 mg, 0.12 mmol, 1.2 eq), zinc powder (1.5 mg, 0.01 mmol, 0.1 eq), tris(dibenzylideneacetone)dipalladium (9.1 mg, 0.01 mmol, 0.1 eq), and 1,1-bis(diphenylphosphino)ferrocene (11 mg, 0.02 mmol, 0.2 eq) were added sequentially. The mixture was stirred at 120°C under argon for 3 h. LC-MS monitoring of the reaction was performed. After completion, the reaction was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to yield 5 mg of the title compound. ESI-MS m / z: 588.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ8.29(s,1H),8.08-8.02(m,3H),7.69(d,J=8.0Hz,1H),7.46-7.42(m,1H),7.13(d,J=8.0Hz,1H),5.22(s,2H),4.32- 4.24(m,2H),4.01(s,2H),3.89(s,2H),3.21(d,J=4.0Hz,2H),3.12-3. 05(m,2H),2.91-2.81(m,2H),2.61-2.53(m,4H),1.09(d,J=4.0Hz,3H).

[0185] Example 11: 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-(3-methyl-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0186]

[0187] Step 1: Preparation of 3-methyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole

[0188]

[0189] Dissolve tert-butyl 3-methyl-2,6-dihydropyrrolo[3,4-c]pyrazole-5(4H)-carboxylate (0.12 g, 0.55 mmol, 1.0 eq) in dichloromethane (4 mL), cool to 0°C in an ice bath, and add trifluoroacetic acid (1.5 mL, 5.5 mmol, 10.0 eq) dropwise. Allow to react at room temperature for 2 h. Monitor the reaction by LC-MS. After completion, the reaction mixture was evaporated to dryness to yield 58 mg of the title compound. ESI-MS m / z: 124.1 [M+H] + .

[0190] Step 2: Preparation of 2-(3-((1s,3s)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-(3-methyl-2,6-dihydropyrrolo[3,4-c]pyrazol-5(4H)-yl)methyl)-4-(trifluoromethyl)isoindol-1-one

[0191]

[0192] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 3-methyl-2,4,5,6-tetrahydropyrrolo[3,4-c]pyrazole to obtain 43 mg of the title compound. ESI-MS m / z: 562.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6): 12.06(s,1H),8.30(s,1H),8.06(d,J=8.0Hz,2H),8.01(s,1H),7.71(d,J=8.0Hz,1H),7.47-7.41(m,1H),7.13(d,J=8.0H z,1H),5.22(s,2H),4.12(s,2H),3.69(s,2H),3.61(s,2H),3.23(s,3H) ,2.87(s,2H),2.57(d,J=4.0Hz,3H),2.14(s,3H),1.10(d,J=4.0Hz,3H).

[0193] Example 12: 6-((3,4-dihydroisoquinolin-2(1H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one

[0194]

[0195] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 1,2,3,4-tetrahydroisoquinoline to obtain 15 mg of the title compound. ESI-MS m / z: 572.3 [M+H] + . 1 HNMR(400MHz,DMSO-d6):8.29(s,1H),8.07-8.05(m,2H),8.00(s,1H),7. 90(s,1H),7.69(m,1H),7.46-7.40(m,1H),7.12-7.08(m,4H),7.02(d,J=4 .0Hz,1H),5.21(s,2H),3.88(s,2H),3.60(s,1H),3.22(s,3H),2.88-2.8 2(m,4H),2.74(d,J=8.0Hz,2H),2.58-2.54(m,3H),1.09(d,J=4.0Hz,3H).

[0196] Example 13: N-cyclopropyl-7-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxamide

[0197]

[0198] Step 1: Preparation of tert-butyl 4-amino-3-oxopiperazine-1-carboxylate

[0199]

[0200] Dissolve tert-butyl 3-oxopiperazine-1-carboxylate (1.0 g, 5.0 mmol, 1.0 eq) in N,N-dimethylacetamide (15 mL). After cooling to 0°C, add sodium hydride (0.3 g, 7.5 mmol, 1.5 eq). Stir for 20 min, then add (aminooxy)diphenylphosphine oxide (1.5 g, 6.5 mmol, 1.3 eq). Stir overnight at room temperature. Monitor the reaction by LC-MS. After completion, quench with water and extract with dichloromethane. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, and concentrate to yield 1.1 g of the title compound.

[0201] Step 2: Preparation of tert-butyl 4-(2-ethoxy-2-oxoacetimido)-3-oxopiperazine-1-carboxylate

[0202]

[0203] Dissolve tert-butyl 4-amino-3-oxopiperazine-1-carboxylate (1.4 g, 6.5 mmol, 1.0 eq) in anhydrous ethanol (15 mL), add ethyl 2-ethoxy-2-iminoacetate (2.36 g, 16.3 mmol, 2.5 eq), and stir at 90°C for 3 h. Monitor the reaction using LC-MS. After completion, quench the reaction with water and extract with dichloromethane. The combined organic phases are washed three times with water, dried over anhydrous sodium sulfate, and concentrated to yield 1.6 g of the title compound. ESI-MS m / z: 315.2 [M+H] + .

[0204] Step 3: Preparation of ethyl 7-(tert-butoxycarbonyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxylate

[0205]

[0206] Dissolve tert-butyl 4-(2-ethoxy-2-oxoacetimidopoly)-3-oxopiperazine-1-carboxylate (2.0 g, 6.36 mmol, 1.0 eq) in N,N-dimethylformamide (20 mL), add toluenesulfonic acid (1.30 g, 7.63 mmol, 1.2 eq), and stir at 130°C for 2 h. Monitor the reaction by LC-MS. After completion, quench the reaction with water and extract with dichloromethane. The combined organic phases are washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 1.38 g of the title compound. ESI-MS m / z: 297.2 [M+H] + .

[0207] Step 4: Preparation of 7-(tert-Butoxycarbonyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxylic acid

[0208]

[0209] Dissolve ethyl 7-(tert-butoxycarbonyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxylate (0.11 g, 0.37 mmol, 1.0 eq) in anhydrous methanol (3.5 mL). Add water (3.5 mL) and sodium hydroxide (0.15 g, 3.7 mmol, 10.0 eq) and stir at room temperature for 1 hour. Monitor the reaction by LC-MS. After completion, adjust the pH to 5 with dilute hydrochloric acid and extract with ethyl acetate. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, and concentrate to yield 0.06 g of the title compound. ESI-MS m / z: 269.2 [M+H] + .

[0210] Step 5: Preparation of tert-butyl 2-(cyclopropylcarbamoyl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate

[0211]

[0212] 7-(tert-Butoxycarbonyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxylic acid (60 mg, 0.22 mmol, 1.0 eq) was dissolved in dichloromethane (3.6 mL). Cyclopropylamine (15.5 mg, 0.27 mmol, 1.2 eq), 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (167.5 mg, 0.44 mmol, 2.0 eq), and N,N-diisopropylethylamine (188.0 mg, 0.88 mmol, 4.0 eq) were added sequentially. The mixture was stirred at room temperature under argon for 1 h. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 195 mg of the title compound. ESI-MS m / z:308.2[M+H] + .

[0213] Step 6: Preparation of N-cyclopropyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxamide

[0214]

[0215] Dissolve tert-butyl 2-(cyclopropylcarbamoyl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate (0.20 g, 0.63 mmol, 1.0 eq) in dichloromethane (5 mL), cool to 0°C in an ice bath, and add trifluoroacetic acid (2.0 mL, 6.35 mmol, 10.0 eq) dropwise. Allow to react at room temperature for 2 h. Monitor the reaction by LC-MS. After completion, slowly add saturated sodium bicarbonate solution dropwise in an ice bath, adjust the pH to 8, and extract with dichloromethane. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, and concentrate to yield 0.18 g of the title compound. ESI-MS m / z: 208.1 [M+H] + .

[0216] Step 7: Preparation of N-cyclopropyl-7-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxamide

[0217]

[0218] 6-(Chloromethyl)-2-(3-((1s,3s)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindol-1-one (0.23 g, 0.48 mmol, 1.0 eq) was dissolved in dioxane (4 mL), and N-cyclopropyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxylate was added in sequence. The mixture was stirred at 100°C for 8 h under argon atmosphere with the addition of (2-dicyclohexylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) methanesulfonate (43.5 mg, 0.05 mmol, 0.1 eq), and potassium carbonate (0.2 g, 1.44 mmol, 3.0 eq). The reaction was monitored by LC-MS. After completion, the mixture was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to yield 34 mg of the title compound. ESI-MS m / z: 646.3 [M+H] + . 1 H NMR(400MHz,DMSO-d6):8.45(d,J=4.0Hz,1H),8.29(s,1H),8.07-8.03(m,3H ),7.71-7.69(m,1H),7.45-7.42(m,1H),7.15-7.13(m,1H),5.22(s,2H),4.2 1-4.18(m,2H),4.01(s,2H),3.84(s,2H),3.22(s,3H),3.06-3.03(m,2H),2. 90-2.79(m,3H),2.61-2.52(m,3H),1.09(d,J=4.0Hz,3H),0.68-0.56(m,4H).

[0219] Example 14: 2-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-1,2,3,4-tetrahydroisoquinoline-7-carbonitrile

[0220]

[0221] The preparation method was the same as that of Example 13, except that the starting material N-cyclopropyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxamide was replaced with 1,2,3,4-tetrahydroisoquinoline-7-carbonitrile to obtain 34 mg of the title compound. ESI-MS m / z: 597.3 [M+H] + . 1 H NMR(400MHz,DMSO-d6):8.30(s,1H),8.09-8.05(m,2H),7.99(s,1H),7.70- 7.68(m,1H),7.61-7.54(m,2H),7.45-7.42(m,1H),7.33(d,J=8.0Hz,1H),7. 13(d,J=8.0Hz,1H),5.22(s,2H),3.90(s,2H),3.65(s,2H),3.22(s,3H),2.9 5-2.83(m,4H),2.78-2.75(m,2H),2.61-2.53(m,3H),1.09(d,J=4.0Hz,3H).

[0222] Example 15: 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-((2-(thiazol-5-yl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindolin-1-one

[0223]

[0224] The compound from Example 9 (40.0 mg, 0.06 mmol, 1.0 eq) was dissolved in anhydrous dioxane (8 mL). 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole (16.0 mg, 0.08 mmol, 1.2 eq), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (2.28 mg, 0.003 mmol, 0.05 eq), cesium carbonate (60.4 g, 0.19 mmol, 3.0 eq), and water (0.8 mL) were added sequentially. The mixture was stirred at 90°C under argon for 4 h. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to yield 9 mg of the title compound. ESI-MS m / z: 646.2 [M+H]. + . 1 H NMR (400MHz, DMSO-d6): δ9.13(d,J=4.0Hz,1H),8.31(d,J=4.0Hz,1H),8.29(s,1H), 8.09(s,1H),8.07-8.05(m,1H),8.04(s,1H),7.72-7.68(m,1H),7.46-7.42(m,1H),7 .13-7.11(m,1H),5.22(s,2H),4.23-4.19(m,2H),4.02(s,2H),3.86(s,2H),3.22(s, 3H),3.08-3.04(m,2H),2.88-2.84(m,2H),2.57-2.53(m,3H),1.09(d,J=8.0Hz,3H).

[0225] Example 16: 2-(3-((1S,3R)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)6-(((S)-3-methylpiperidin-1-yl)methyl)-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one

[0226]

[0227] Step 1: Preparation of (S)-6-((3-methylpiperidin-1-yl)methyl)-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one

[0228]

[0229] 6-Bromo-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (0.1 g, 0.35 mmol, 1.0 eq) was dissolved in dioxane (10 mL), and (S)-3-methyl-1-((trifluoro-4-boryl)methyl)piperidinium potassium salt (0.23 g, 1.06 mmol, 3.0 eq), methanesulfonic acid (2-dicyclohexylphosphine-2',4',6' 2-(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (0.06 g, 0.07 mmol, 0.2 eq), 2-dicyclohexylphosphino-2',4',6'-triisopropylbiphenyl (0.07 g, 0.14 mmol, 0.4 eq), potassium carbonate (0.2 g, 1.42 mmol, 4.0 eq), and water (2.5 mL) were stirred overnight at 90°C under argon protection. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 79 mg of the title compound. ESI-MS m / z: 315.1 [M+H] + .

[0230] Step 2: Preparation of 2-(3-((1S,3R)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-(((S)-3-methylpiperidin-1-yl)methyl)-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one

[0231]

[0232] (S)-6-((3-methylpiperidin-1-yl)methyl)-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one (65 mg, 0.21 mmol, 1.0 eq) was dissolved in N,N-dimethylformamide (8 mL), and 3-((1s,3s)-1-(3-bromophenyl)-3-methylcyclobutyl)-4-methyl-4H-1,2 ,4-triazole (63 mg, 0.21 mmol, 1.0 eq), cuprous iodide (8 mg, 0.042 mmol, 0.20 eq), (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (12 mg, 0.084 mmol, 0.40 eq) and potassium carbonate (87 mg, 0.63 mmol, 3.00 eq) were stirred at 100°C for 3 h under argon protection. The reaction was monitored by LC-MS. After completion of the reaction, water was added to quench the reaction, and dichloromethane was added for extraction. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to obtain 28 mg of the title compound. ESI-MS m / z: 540.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ8.31(s,1H),8.08(s,1H),8.01-7.97(m,1H),7.86(dd,J =8.0Hz,1H),7.78(s,1H),7.58-7.54(m,1H),7.37(d,J=8.0Hz,1H),3.51(s,2H), 3.20(s,3H),2.85-2.82(m,4H),2.59(d,J=4.0Hz,3H),2.03-1.99(m,1H),1.67-1 .63(m,4H),1.49(d,J=12.0Hz,2H),1.10(d,J=4.0Hz,3H),0.84(d,J=8.0Hz,3H).

[0233] Example 17: 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-((2-(pyridin-3-yl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindolin-1-one

[0234]

[0235] The preparation method was the same as that of Example 15, except that the starting material 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)thiazole was replaced with 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine to obtain 13 mg of the title compound. ESI-MS m / z: 641.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6):9.14(s,1H),8.61(m,1H),8.28-8.26(m,1H),8.10(s, 1H),8.06-8.05(m,2H),7.11-7.09(m,1H),7.52-7.42(m,2H),7.14(d,J=8.0Hz ,1H),5.23(s,2H),4.27-4.21(m,2H),4.04(s,2H),3.88(s,2H),3.22(s,3H),3 .11-3.07(m,2H),2.87-2.86(m,2H),2.57-2.55(m,3H),1.09(d,J=4.0Hz,3H).

[0236] Example 18: 7-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine-2-carbonitrile

[0237]

[0238] The preparation method was the same as that of Example 7, except that the starting material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 5,6,7,8-tetrahydroimidazo[1,2-a]pyrazine-2-carbonitrile to obtain 15 mg of the title compound. ESI-MS m / z: 587.3 [M+H] + . 1 H NMR(400MHz, DMSO-d6):8.31(s,1H),8.11-8.03(m,3H),8.01(s,1H),7.72-7.68(mz,1H),7.50-7.40(m,1H),7.14(d,J=8.0Hz,1H),5.22(s,2 H),4.09-4.05(m,2H),3.96(s,2H),3.70(s,2H),3.22(s,3H),2.96-2. 92(m,2H),2.90-2.82(m,2H),2.61-2.53(m,3H),1.10(d,J=4.0Hz,3H).

[0239] Example 19: 6-((2-ethoxy-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0240]

[0241] Step 1: Preparation of tert-butyl 2-ethoxy-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate

[0242]

[0243] Dissolve tert-butyl 2-bromo-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate (0.13 g, 0.43 mmol, 1.0 eq) in ethanol (4 mL) and dioxane (8 mL), and add tris(dibenzylideneacetone)dipalladium(0) (0.06 g, 0.07 mmol, 0.16 eq), 2-di-tert-butylphosphino-2',4',6'-triisopropyl-3,6-dimethoxy-1,1'-biphenyl (0.04 g, 0.079 mmol, 1.1 eq) and potassium hydroxide (0.08 g, 1.43 mmol, 3.3 eq) in sequence. Stir at 100°C under argon protection overnight. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 0.11 g of the title compound. ESI-MS m / z: 269.2 [M+H] + .

[0244] Step 2: Preparation of 2-ethoxy-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine

[0245]

[0246] Dissolve tert-butyl 2-ethoxy-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate (56 mg, 0.21 mmol, 1.00 eq) in dichloroethane (2 mL), cool to 0°C in an ice bath, add trifluoroacetic acid (0.5 mL, 6.73 mmol, 32.2 eq) dropwise, and stir at 70°C for 1 h. Monitor the reaction by LC-MS. After completion, slowly add saturated sodium bicarbonate solution dropwise in an ice bath, adjust the pH to 8, and extract with dichloromethane. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, and concentrate to yield 34 mg of the title compound. ESI-MS m / z: 169.2 [M+H] + .

[0247] Step 3: Preparation of 6-((2-ethoxy-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0248]

[0249] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 2-ethoxy-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine to obtain 8 mg of the title compound. ESI-MS m / z: 607.3 [M+H] + . 1 HNMR (400MHz, DMSO-d6): 8.31 (s, 1H), 8.15-7.99 (m, 3H), 7.70 (d, J = 8.0Hz, 1H), 7.49-7.39 (m, 1H), 7.14 (d, J = 8. 0Hz,1H),5.22(s,2H),4.20-4.10(m,2H),4.02-3.90(m,3H),3.66(s,2H),3.23(s,3H),3.01-2.97(m,2H),2.90- 2.80 (m, 2H), 2.62-2.52 (m, 3H), 1.31-1.27 (m, 3H), 1.10 (d, J = 4.0 Hz, 3H). Example 20: 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-((2-(methylsulfonyl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindolin-1-one

[0250]

[0251] Step 1: Preparation of tert-butyl 2-bromo-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate

[0252]

[0253] Dissolve 2-bromo-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine (0.9 g, 4.45 mmol, 1.0 eq) in anhydrous dichloromethane (10 mL). Add di-tert-butyl dicarbonate (1.16 g, 5.34 mmol, 1.2 eq) and triethylamine (0.59 g, 5.78 mmol, 1.3 eq) sequentially. Stir under argon at room temperature for 5 h. Monitor the reaction by LC-MS. After completion, quench with water and extract with dichloromethane. The combined organic phases are washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 1.2 g of the title compound. ESI-MS m / z: 303.1 [M+H]. + .

[0254] Step 2: Preparation of tert-butyl 2-(methylsulfonyl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate

[0255]

[0256] Dissolve tert-butyl 2-bromo-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate (0.15 g, 0.5 mmol, 1.0 eq) in anhydrous dimethyl sulfoxide (3 mL). Add sodium methanesulfinate (76.5 mg, 0.75 mmol, 1.5 eq), copper trifluoromethanesulfonate (36.2 mg, 0.10 mmol, 0.2 eq), and (1R,2R)-(-)-N,N'-dimethyl-1,2-cyclohexanediamine (28.5 mg, 0.20 mmol, 0.4 eq) sequentially. Stir under argon at 110°C for 8 h. Monitor the reaction by LC-MS. After completion, quench the reaction with water and extract with dichloromethane. The combined organic phases are washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to yield 87 mg of the title compound. ESI-MS m / z:303.1[M+H] + Step 3: Preparation of 2-(methylsulfonyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine

[0257]

[0258] Dissolve tert-butyl 2-(methylsulfonyl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate (87 mg, 0.28 mmol, 1.0 eq) in dichloromethane (5 mL), cool to 0°C in an ice bath, add trifluoroacetic acid (1.0 mL, 2.88 mmol, 10.0 eq) dropwise, and stir at room temperature for 1 hour. Monitor the reaction by LC-MS. After completion, slowly add saturated sodium bicarbonate solution dropwise in an ice bath, adjust the pH to 8, and extract with dichloromethane. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, and concentrate to yield 64 mg of the title compound. ESI-MS m / z: 203.1 [M+H] + .

[0259] Step 4: Preparation of 2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-6-((2-(methylsulfonyl)-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-4-(trifluoromethyl)isoindolin-1-one

[0260]

[0261] The preparation method was the same as that of Example 13, except that the starting material N-cyclopropyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine-2-carboxamide was replaced with 2-(methylsulfonyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine to obtain 5 mg of the title compound. ESI-MS m / z: 641.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6):8.29(s,1H),8.08-8.03(m,3H),7.71-7.68(m,1H),7.46-7.42(m,1H),7.14(d,J=8.0Hz,1H),5.22(s,2H),4.29-4.27 (m,2H),4.02(s,2H),3.91(s,2H),3.32(s,3H),3.22(s,3H),3.11-3.0 8(m,2H),2.90-2.83(m,2H),2.59-2.55(m,3H),1.09(d,J=4.0Hz,3H).

[0262] Example 21: 2-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindolin-5-yl)methyl)isoquinoline-5-carbonitrile

[0263]

[0264] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with isoindoline-5-carbonitrile hydrochloride to obtain 28 mg of the title compound. ESI-MS m / z: 583.2 [M+H] + . 1 HNMR(400MHz,DMSO-d6):8.30(s,1H),8.08-8.04(m,2H),8.02(s,1H),7.74-7.68(m,3H),7.50-7.42(m,2H),7.17-7.11(m,1H) ),5.23(s,2H),4.11(s,2H),3.97(d,J=12.0Hz,4H),3.22(s,3H),2.91-2.81(m,2H),2.60-2.54(m,3H),1.09(d,J=4.0Hz,3H).

[0265] Example 22: 2'-(3-((1S,3R)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-5'-(((S)-3-methylpiperidin-1-yl)methyl)spiro[cyclopropane-1,1'-isoindolin]-3'-one

[0266]

[0267] Step 1: Preparation of 5'-bromospiro[cyclopropane-1,1'-isoindolin]-3'-one

[0268]

[0269] Methyl 5-bromo-2-cyanobenzoate (2.56 g, 10.6 mmol, 1.0 eq) was dissolved in anhydrous tetrahydrofuran (60 mL). Under argon, tetraisopropyl titanate (4.25 g, 15.0 mmol, 1.4 eq) and ethylmagnesium bromide (13.3 mL, 26.6 mmol, 2.5 eq) were added sequentially. Stirring was performed at room temperature for 1.5 h. The reaction was monitored by LC-MS. After completion, the mixture was quenched with saturated ammonium chloride at 0°C and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 100 mg of the title compound. ESI-MS m / z: 238.0 [M+H] + .

[0270] Step 2: Preparation of (S)-5'-((3-methylpiperidin-1-yl)methyl)spiro[cyclopropane-1,1'-isoindolin]-3'-one

[0271]

[0272] 5'-Bromospiro[cyclopropane-1,1'-isoindolin]-3'-one (45 mg, 0.19 mmol, 1.0 eq) was dissolved in dioxane (4 mL), and (S)-3-methyl-1-((trifluoro-4-boryl)methyl)piperidinium potassium salt (45 mg, 0.21 mmol, 1.1 eq), methanesulfonic acid (2-dicyclohexylphosphino-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (7 mg, 0.008 mmol, 0.04 eq), potassium carbonate (230 mg, 1.67 mmol, 8.84 eq) and water (0.4 mL) were added in sequence. The mixture was stirred at 80°C under argon protection overnight. The reaction was monitored by LC-MS. After completion, the mixture was quenched with water and extracted with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 30 mg of the title compound. ESI-MS m / z: 271.2 [M+H] + .

[0273] Step 3: Preparation of 2'-(3-((1S,3R)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-5'-(((S)-3-methylpiperidin-1-yl)methyl)spiro[cyclopropane-1,1'-isoindolin]-3'-one

[0274]

[0275] The preparation method was the same as that of Example 16, except that the starting material (S)-6-((3-methylpiperidin-1-yl)methyl)-8-(trifluoromethyl)-[1,2,4]triazolo[4,3-a]pyridin-3(2H)-one was replaced with (S)-5'-((3-methylpiperidin-1-yl)methyl)spiro[cyclopropane-1,1'-isoindolin]-3'-one to obtain 6 mg of the title compound. ESI-MS m / z: 496.3 [M+H] + . 1H NMR(400MHz,DMSO-d6):8.31(s,1H),7.69(s,1H),7.60-7.56(m,1H),7.54-7.48(m,1H),7.39( d,J=8.0Hz,1H),7.34(d,J=8.0Hz,1H),7.2-7.11(m,2H),3.60-3.56(m,2H),3.2(s,3H),2.90-2 .79(m,2H),2.78-2.70(m,2H),2.59-2.52(m,3H),1.93-1.89(m,1H),1.70-1.48(m,5H),1.48-1 .40(m,2H),1.26-1.12(m,2H),1.08(d,J=4.0Hz,3H),0.91-0.83(m,1H),0.82(d,J=4.0Hz,3H).

[0276] Example 23: N-(7-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindolin-5-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)acetamide

[0277]

[0278] Step 1: Preparation of N-([1,2,4]triazolo[1,5-a]pyrazin-2-yl)acetamide

[0279]

[0280] Dissolve [1,2,4]triazolo[1,5-a]pyrazin-2-amine (0.8 g, 5.92 mmol, 1.0 eq) in anhydrous dioxane (10 mL), add acetic anhydride (1.5 g, 14.81 mmol, 2.5 eq), and stir overnight at 110°C under argon. Monitor the reaction by LC-MS. After completion, filter and dry the filter cake to obtain 0.61 g of the title compound. ESI-MS m / z: 178.1 [M+H] + .

[0281] Step 2: Preparation of N-(5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)acetamide

[0282]

[0283] Dissolve N-([1,2,4]triazolo[1,5-a]pyrazin-2-yl)acetamide (0.6 g, 3.39 mmol, 1.0 eq) in anhydrous ethanol (10 mL). Add platinum dioxide (0.12 g, 0.51 mmol, 0.15 eq). Replace the atmosphere with hydrogen and stir overnight at room temperature. Monitor the reaction by LC-MS. After completion, filter the mixture, dry, and concentrate the filtrate to obtain 0.58 g of the title compound. ESI-MS m / z: 182.1 [M+H] + .

[0284] Step 3: Preparation of N-(7-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindolin-5-yl)methyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)acetamide

[0285]

[0286] The preparation method was the same as that of Example 7, except that the starting material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with N-(5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazin-2-yl)acetamide to obtain 35 mg of the title compound. ESI-MS m / z: 620.3 [M+H] + . 1 H NMR(400MHz,DMSO-d6):10.22(s,1H),8.29(s,1H),8.08-8.04(m,2H),8.02( s,1H),7.72-7.68(m,1H),7.47-7.42(m,1H),7.15-7.11(m,1H),5.22(s,2H) ,4.08-4.02(m,2H),3.98(s,2H),3.73(s,2H),3.22(s,3H),3.03-2.98(m,2H ),2.90-2.82(m,2H),2.57-2.53(m,3H),2.01(s,3H),1.09(d,J=8.0Hz,3H).

[0287] Example 24: 6-((2-cyclopropyl-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0288]

[0289] Step 1: Preparation of 2-cyclopropyl-[1,2,4]triazolo[1,5-a]pyrazine

[0290]

[0291] 2-Bromo-[1,2,4]triazolo[1,5-a]pyrazine (0.5 g, 2.53 mmol, 1.0 eq) was dissolved in dioxane (20 mL). Cyclopropylboronic acid pinacol ester (0.42 g, 2.53 mmol, 1.0 eq), 1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.19 g, 0.26 mmol, 0.1 eq), potassium trimethylsilanol (1.30 g, 10.12 mmol, 4.0 eq), and water (2 mL) were added sequentially. The mixture was stirred at 100°C overnight under argon. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 110 mg of the title compound. ESI-MS m / z: 161.1 [M+H]. + .

[0292] Step 2: Preparation of 2-cyclopropyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine

[0293]

[0294] Dissolve 2-cyclopropyl-[1,2,4]triazolo[1,5-a]pyrazine (50 mg, 0.31 mmol, 1.0 eq) in anhydrous methanol (2 mL), cool to 0°C in an ice bath, and add a solution of lithium borohydride in tetrahydrofuran (0.6 mL, 1.2 mmol, 4.0 eq) dropwise. Stir at room temperature for 4 h. Monitor the reaction by LC-MS. After completion, quench with water, remove the solvent by rotary evaporation, dilute with water, extract with dichloromethane, and combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, concentrate, and purify by column chromatography to obtain 36 mg of the title compound. ESI-MS m / z: 165.1 [M+H] + Step 3: Preparation of 6-((2-cyclopropyl-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0295]

[0296] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 2-cyclopropyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine to obtain 5 mg of the title compound. ESI-MS m / z: 603.3 [M+H] + . 1 HNMR(400MHz,DMSO-d6):8.29(s,1H),8.05(s,2H),8.00(s,1H),7.69(d,J=8.0Hz,1H ),7.45-7.42(m,1H),7.13(d,J=8.0Hz,1H),5.21(s,2H),4.04-4.01(m,2H),3.96(s,2 H),3.68(s,2H),3.22(s,3H),2.98-2.96(m,2H),2.87-2.84(m,2H),2.56(d,J=4.0Hz ,3H),2.03-1.97(m,1H),1.09(d,J=4.0Hz,3H),0.87-0.84(m,2H),0.75-0.73(m,2H).

[0297] Example 25: 6-((2-cyclopropylethynyl-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0298]

[0299] Step 1: Preparation of tert-butyl 2-cyclopropylethynyl-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate

[0300]

[0301] Dissolve tert-butyl 2-bromo-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate (0.21 g, 0.69 mmol, 1.0 eq) in N,N-diisopropylethylamine (6 mL). Add cyclopropylacetylene (0.15 g, 2.27 mmol, 3.28 eq), tetrakistriphenylphosphine palladium(0) (91 mg, 0.079 mmol, 0.11 eq), and cuprous iodide (32 mg, 0.17 mmol, 0.24 eq). Stir overnight at 100°C under argon. Monitor the reaction by LC-MS. After completion, quench with water and extract with dichloromethane. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, concentrate, and purify by column chromatography to obtain 45 mg of the title compound. ESI-MS m / z: 289.2 [M+H]. + .

[0302] Step 2: Preparation of 2-cyclopropylethynyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine

[0303]

[0304] Dissolve tert-butyl 2-cyclopropylethynyl-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazine-7(8H)-carboxylate (40 mg, 0.14 mmol, 1.0 eq) in dichloromethane (1 mL), cool to 0°C in an ice bath, and add trifluoroacetic acid (0.11 mL, 1.4 mmol, 10.0 eq) dropwise. Allow to react at room temperature for 2 h. Monitor the reaction by LC-MS. After completion, slowly add saturated sodium bicarbonate solution dropwise in an ice bath, adjust the pH to 8, and extract with dichloromethane. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, and concentrate to yield 42 mg of the title compound. ESI-MS m / z: 189.4 [M+H] + .

[0305] Step 3: Preparation of 6-((2-cyclopropylethynyl-5,6-dihydro-[1,2,4]triazolo[1,5-a]pyrazin-7(8H)-yl)methyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindolin-1-one

[0306]

[0307] The preparation method was the same as that of Example 7, except that the raw material 4,5,6,7-tetrahydro-1H-pyrrolo[3,2-c]pyridine was replaced with 2-cyclopropylethynyl-5,6,7,8-tetrahydro-[1,2,4]triazolo[1,5-a]pyrazine to obtain 13 mg of the title compound. ESI-MS m / z: 627.3 [M+H] + . 1H NMR(400MHz, DMSO-d6):8.30(s,1H),8.09-8.04(m,2H),8.02(s,1H),7.70(d,J=8.0H z,1H),7.48-7.41(m,1H),7.14(d,J=8.0Hz,1H),5.22(s,2H),4.14-4.07(m,2H),3.98 (s,2H),3.76(s,2H),3.22(s,3H),3.04-2.98(m,2H),2.92-2.81(m,2H),2.61-2.53(m ,3H),1.60-1.52(m,1H),1.10(d,J=4.0Hz,3H),0.94-0.87(m,2H),0.78-0.71(m,2H).

[0308] Example 26: 5-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-2-carbonitrile

[0309]

[0310] Step 1: Preparation of tert-butyl 2-iodo-3-((2-(trimethylsilyl)ethoxy)methyl)-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylate

[0311]

[0312] Dissolve tert-butyl 2-iodo-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylate (0.5 g, 1.27 mmol, 1.0 eq) in anhydrous tetrahydrofuran (15 mL) and cool to 0°C in an ice bath. Add sodium hydride (56 mg, 1.40 mmol, 1.1 eq) and stir at room temperature for 1.5 h. Then, add 2-(trimethylsilyl)ethoxymethyl chloride (233 mg, 1.40 mmol, 1.1 eq) in an ice bath and stir at room temperature overnight. Monitor the reaction by LC-MS. After completion, quench with water and extract with dichloromethane. Combine the organic phases, wash three times with water, dry over anhydrous sodium sulfate, concentrate, and purify by column chromatography to obtain 0.8 g of the title compound. ESI-MS m / z: 480.1 [M+H]. + .

[0313] Step 2: Preparation of tert-butyl 2-cyano-3-((2-(trimethylsilyl)ethoxy)methyl)-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylate

[0314]

[0315] Dissolve tert-butyl 2-iodo-3-((2-(trimethylsilyl)ethoxy)methyl)-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylate (0.48 g, 1.0 mmol, 1.0 eq) in N,N-dimethylacetamide (8 mL), and add zinc cyanide (0.14 g, 1.20 mmol, 1.2 eq), zinc powder (7 mg, 0.10 mmol, 0.1 eq), 1,1'-bis(diphenylphosphino)ferrocene (0.11 g, 0.20 mmol, 0.2 eq) and tris(dibenzylideneacetone)dipalladium (92 mg, 0.10 mmol, 0.1 eq) in sequence. Under argon protection, stir at 120 °C for 4 h. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 0.12 g of the title compound. ESI-MS m / z: 379.2 [M+H] + .

[0316] Step 3: Preparation of 3-((2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-2-carbonitrile

[0317]

[0318] Dissolve tert-butyl 2-cyano-3-((2-(trimethylsilyl)ethoxy)methyl)-3,4,6,7-tetrahydro-5H-imidazo[4,5-c]pyridine-5-carboxylate (90 mg, 0.24 mmol, 1.0 eq) in dichloromethane (3 mL), cool to 0°C in an ice bath, and add trifluoroacetic acid (0.3 mL, 3.8 mmol, 16.0 eq) dropwise. Allow to react at room temperature for 3 h. Monitor the reaction by LC-MS. After completion, slowly add saturated sodium bicarbonate solution dropwise in an ice bath, adjust the pH to 8, and extract with dichloromethane. The organic phases are combined, washed three times with water, dried over anhydrous sodium sulfate, and concentrated to yield 60 mg of the title compound. ESI-MS m / z: 279.2 [M+H] + .

[0319] Step 4: Preparation of 5-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-3-(2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-2-carbonitrile

[0320]

[0321] 3-((2-(Trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-2-carbonitrile (60 mg, 0.22 mmol, 1.0 eq) was dissolved in dioxane (5 mL), and 6-(chloromethyl)-2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-4-(trifluoromethyl)isoindole was added successively. Indole-1-one (60 mg, 0.13 mmol, 0.6 eq), methanesulfonic acid (2-dicyclohexylphosphino)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl) palladium(II) (30 mg, 0.033 mmol, 0.15 eq), and cesium carbonate (0.25 g, 0.77 mmol, 3.5 eq) were stirred at 100°C for 4.5 hours under argon protection. The reaction was monitored by LC-MS. After completion, the reaction was quenched with water and extracted with ethyl acetate. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by column chromatography to obtain 40 mg of the title compound. ESI-MS m / z: 718.9 [M+H] + .

[0322] Step 5: Preparation of 5-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-2-carbonitrile

[0323]

[0324] 5-((2-(3-((1S,3S)-3-methyl-1-(4-methyl-4H-1,2,4-triazol-3-yl)cyclobutyl)phenyl)-3-oxo-7-(trifluoromethyl)isoindol-5-yl)methyl)-3-(2-(trimethylsilyl)ethoxy)methyl)-4,5,6,7-tetrahydro-3H-imidazo[4,5-c]pyridine-2-carbonitrile (35 mg, 0.049 mmol, 1.0 eq) was dissolved in dichloromethane (5 mL), cooled to 0°C in an ice bath, and trifluoroacetic acid (1.0 mL) was added dropwise. The reaction was allowed to react at room temperature for 4 h. The reaction was monitored by LC-MS. After completion of the reaction, saturated sodium bicarbonate solution was slowly added dropwise in an ice bath to adjust the pH to 8. The mixture was extracted with dichloromethane. The organic phases were combined, washed three times with water, dried over anhydrous sodium sulfate, concentrated, and purified by C18 column chromatography to afford 5 mg of the title compound. ESI-MS m / z:588.5[M+H] + . 1 H NMR (400MHz, DMSO-d6): δ8.29(s,1H),8.04(d,J=8.0Hz,2H),7.98(s,1H),7.71-7.67(m,1H),7.46-7.42(m,1H),7.12(d,J=8.0Hz,1H),5.21(s,2 H),3.92(s,2H),3.49(s,2H),3.21(s,3H),2.90-2.84(m,2H),2.83-2.76 (m,2H),2.67-2.65(m,2H),2.56(d,J=4.0Hz,3H),1.09(d,J=8.0Hz,3H).

[0325] Experimental Example 1 Evaluation of Cbl-b activity of compounds in vitro

[0326] 1. Experimental Materials

[0327] Compounds: The compounds of the present invention prepared in the above examples were each prepared with DMSO to 30 mM and then diluted to 10000 nM, 2500 nM, 625 nM, 156.25 nM, 39.06 nM, 9.77 nM, 2.44 nM, 0.61 nM, 0.15 nM, and 0.038 nM, respectively.

[0328] Reagents and consumables: Cbl-b, GST-Tag (Human) Recombinant, purchased from BPS, Cat. No. 80415; UBE1 (UBA1), FLAG-tag Recombinant, purchased from BPS, Cat. No. 80301; UbcH5b, His-Tag (Human) Recombinant, purchased from BPS, Cat. No. 80314; Biotin-Ubiquitin, purchased from Thermo Fisher Scientific, Cat. No. PV4380; ATP, purchased from Promega, Cat. No. V915B; MAb Anti-GST-Tb cryptate, purchased from Cisbio, Cat. No. 61GSTTLA; Streptavidin-XL665, purchased from Cisbio, Cat. No. 610SAXLA A; HEPES, purchased from Life Sciences. Technologies Company, Cat.No.15630-080; DMSO, purchased from MP, Cat.No.196055.

[0329] Instruments: PHERAstar FSX, purchased from BMG LABTECH, Germany; ECHO, purchased from Labcyte, USA.

[0330] 2. Experimental Methods

[0331] Prepare 1X buffer: 50 mM HEPES, 5 mM MgCl2, 100 mM NaCl, 0.01% Triton X-100, pH 7.0.

[0332] 2.2. Serially dilute the compound stock solution with DMSO and add 0.06 μL of the diluted compound solution to the assay plate using an Echo. Repeat twice for each concentration.

[0333] Dilute UBE1 to 25 ng / μL in assay buffer (batch dependent)

[0334] Dilute UBCH5b to 220 ng / μL in assay buffer (batch dependent)

[0335] Dilute CBLB to 3 ng / μL in U2 assay buffer.

[0336] Dilute Biotin-Ub to 50 ng / μL in U2 assay buffer

[0337] 2.3. Prepare a mixture in each well: 1 μL Biotin-Ub + 1 μL diluted UBE1 + 1 μL diluted UBCH5b + 2.5 μL diluted CBL-B + 2 μL assay buffer, a total of 7.5 μL;

[0338] Positive control (MAX) wells: DMSO.

[0339] Negative control (min) well 1: without Cbl-b

[0340] Negative control well 2: without Biotin-Ub

[0341] Centrifuge the assay plate at 1000 rpm for 1 minute, then dilute ATP to a 40 μM solution and add 2.5 μL to each well.

[0342] Centrifuge the plate again at 1000 rpm for 1 minute and incubate at 25°C for 3 hours.

[0343] Dilute MAb Anti GST-Tb cryptate (1:200) and Streptavidin-XL665 (1:200) in assay buffer.

[0344] 2.7. Add 10 μL of diluted donor / acceptor mixture to each well, incubate at room temperature for 1 hour, and then read the 665 / 620 ratio using a PHERAstar FSX;

[0345] 2.8. Inhibition rate of compound (% inhibition rate) = 100 × (average value of positive control wells - number of compound wells) / (average value of positive control wells - average value of negative control wells 1). The data were processed by XLfit and fitted to obtain IC50 and IC 50 IC of the compound 50 The results are shown in Table 1.

[0346] Table 1

[0347]

[0348]

[0349] From the above experimental results, it can be seen that the compound of the present invention has good inhibitory activity against Cbl-b and is very promising to become a cancer immunotherapy drug with higher efficacy and fewer side effects.

[0350] Experimental Example 2 Evaluation of compound in vitro cell activity

[0351] 1. Experimental Materials

[0352] Test compounds: the compounds of the present invention prepared in the above examples, each compound was prepared with DMSO to 10 mM, and then diluted 3-fold in sequence to 1000.00 nM, 333.33 nM, 111.11 nM, 37.04 nM, 12.35 nM, 4.12 nM, and 1.37 nM.

[0353] Human PBMC cells were purchased from Shanghai Aoneng Biotechnology.

[0354] Reagents: IMDM, purchased from Thermo Fisher Scientific Inc., USA, Cat. NO. 12440053; FBS, purchased from GIBCO, USA, Cat. NO. 10099-141; Penicillin-Streptomycin, purchased from GIBCO, USA, Cat. NO. 15140-122; Anti-Hu CD3, purchased from Invitrogen, USA, Cat. NO. 16-0037-81; Anti-Hu CD28, purchased from Invitrogen, USA, Cat. NO. 16-0289-81; Human IFN-γ ELISA kit, purchased from RD Company, USA, Cat. NO. DY285B.

[0355] Instrument: SpectraMax iD3 / iD5, purchased from Molecular Devices.

[0356] 2. Experimental Methods

[0357] 2.1 Coating of 96-well plates: Coat 96-well plates overnight with Anti-Hu CD3, 2 μg / ml, 100 μL / well, at -4°C overnight.

[0358] 2.2 Wash the plate: Aspirate and discard the liquid in the wells, add 250 μL PBS, shake horizontally, then aspirate and discard the liquid in the wells, repeat three times;

[0359] 2.3 Resuspend the recovered PBMCs, count them, and quantify them to 1×10 6 / mL, then added to a 96-well plate at 100 μL / well

[0360] 2.4 Dilute the compound serially and add it to a 96-well plate to a total of 7 concentrations. Place the 96-well plate in a 37°C incubator. Incubate the cells and compound for 1 hour, then add 50 μl / well of a 4 μg / ml Anti-HuCD28 solution prepared in IMDM complete medium for a final concentration of 1 μg / mL.

[0361] 2.5 Finally, place the 96-well plate in a 37°C, 5% CO2 incubator and incubate for 48 hours.

[0362] 2.6 After incubation for 48 hours, collect the cell culture supernatant by centrifugation;

[0363] 2.7 Detect IL-2 secretion in the cell supernatant using an ELISA kit. Negative control 1 (Min) is the DMSO well, and negative control 2 is the CD3+CD28 well.

[0364] 2.8 EC was performed using GraphicPad 6.0 software according to different drug concentrations and their corresponding IL-2 secretion. 50 Curve drawing, data analysis, and final EC 50 The experimental results are shown in Table 2.

[0365] Table 2

[0366]

[0367] Experimental Example 3 Pharmacokinetic Experiment

[0368] 1. Experimental Materials

[0369] Compound: The compound of the present invention prepared in the above examples, and compound 23 disclosed in WO2020 / 264398 As a control compound (Compound A). The preparation method of Compound A refers to WO2020 / 264398, and it was identified by hydrogen spectrum and mass spectrometry.

[0370] The oral medication is prepared as a 0.5 mg / mL clear solution in a 10% solutol / 20% PG / 70% saline solution (v / v / v). The intravenous medication is prepared as a 0.1 mg / mL clear solution in a 4% PG / 2% solutol / 94% saline solution (v / v / v).

[0371] Animals: Male BALB / c mice, SPF grade, 18-20 g, purchased from Shanghai Slake Laboratory Animal Co., Ltd. They were given a 2-3 day acclimatization period before the experiment.

[0372] Instruments: API4500 triple quadrupole liquid chromatography-mass spectrometer (AB, USA), equipped with an electrospray ionization source (ESI), LC-30AD dual pumps; SIL-30AC automatic sampler; CTO-30AC column oven; DGU-20A3R degasser; AnalystQSA01.01 chromatography workstation; Milli-Q ultrapure water device (Millipore Inc); Qilinbeier Vortex-5 oscillator; HITACHI C F16RⅩⅡ desktop high-speed refrigerated centrifuge.

[0373] 2. Experimental Methods

[0374] (1) Three mice were taken per group and the compound of the present invention and the control compound A were administered orally (IG) at 10 mg / kg each; the compound of the present invention and the control compound A were administered orally (IV) at 1 mg / kg each;

[0375] (2) Blood was collected from the orbital venous plexus at 5 min, 15 min, 30 min, 1 h, 2 h, 6 h, 10 h, and 24 h after intravenous or oral administration into heparinized EP tubes (0.6 mL). After centrifugation at 8000 rpm / min for 5 min, the upper plasma layer was collected and frozen at -20°C for LC-MS / MS analysis.

[0376] (3) Based on the blood drug concentration data obtained in the above steps, a blood drug concentration-time curve was drawn, and the pharmacokinetic parameters were calculated using WinNonlin software.

[0377] 3. Experimental Results

[0378] The experimental results are shown in Tables 3 and 4.

[0379] Table 3 Oral administration

[0380]

[0381] Table 4 Intravenous administration

[0382]

[0383] The experimental results show that the compound of the present invention has a higher AUC in vivo, indicating that the drug exposure in the blood is higher, and a higher bioavailability (F), indicating that the drug is well absorbed and has a better therapeutic effect.

[0384] Although the present invention has been described in detail above, it will be appreciated by those skilled in the art that various modifications and variations may be made to the present invention without departing from the spirit and scope of the present invention. The scope of the present invention is not limited to the detailed description above, but rather to the claims.

Claims

1. A compound represented by general formula (I) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, in, X is selected from C or N; Y is selected from C or N; R 1 、R 2 、R 4 、R 5 each independently selected from hydrogen, halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, and dialkylamino; Or two R 2 and the carbon atoms to which they are attached may together form a spirocycloalkyl group or a spiroheterocyclyl group, wherein the spirocycloalkyl group or the spiroheterocyclyl group is optionally substituted with one or more alkyl groups, alkoxy groups, haloalkoxy groups, haloalkyl groups, halogen groups, hydroxyl groups, carboxyl groups, cyano groups, amino groups, alkylacyl groups, aminoacyl groups, dialkylamino groups and oxo groups; L is selected from a single bond, an alkylene group, an imino group, and an alkylene-imino group; Ring A is selected from aryl, heteroaryl, cycloalkyl and heterocyclyl; R 3 is selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, alkoxy, halogen, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkynyl, alkylsulfonyl, oxo, aminoacyl, alkylacylamino, cycloalkylaminoacyl, wherein the cycloalkyl, heterocyclyl, aryl, heteroaryl is optionally substituted with one or more hydrogen, alkyl, alkoxy, haloalkoxy, haloalkyl, halogen, hydroxy, carboxyl, cyano, amino, alkylacyl, aminoacyl, dialkylamino, and oxo groups; is a single bond or a double bond, two of which All are single keys, or one is a single bond, and the other is a double bond; and m, n and p are each independently selected from 0, 1, 2, and 3.

2. The compound according to claim 1 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein R 1 、R 2 、R 4 、R 5 Each independently selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxy C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl and di-C 1-6 Alkylamino.

3. The compound according to claim 1 or 2, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein L is selected from sub-C 1-6 Alkyl, imino, C 1-6 Alkyl-imino.

4. The compound according to any one of claims 1 to 3, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein ring A is selected from C 6-12 Aryl, 5-15 membered heteroaryl, C 3-12 cycloalkyl and 3-12 membered heterocyclic groups.

5. The compound according to any one of claims 1 to 4, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein R 3 Selected from hydrogen, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, C 1-6 Alkoxy, fluorine, chlorine, bromine, iodine, cyano, C 3-12 Cycloalkyl, 3-12 membered heterocyclic group, C 6-12 Aryl, 5-15 membered heteroaryl, C 3-12 Cycloalkyl C 2-6 Alkynyl, C 1-6 Alkylsulfonyl, oxo, aminoacyl, C 1-6 Alkyl acylamino, C 1-6 Cycloalkylaminoacyl, the C 3-12 Cycloalkyl, 3-12 heterocyclic, C 6-12 Aryl, 5-15 membered heteroaryl optionally substituted with one or more hydrogen, C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, halogenated C 1-6 Alkyl, halogen, hydroxyl, carboxyl, cyano, amino, C 1-6 Alkyl acyl, amino acyl, di-C 1-6 Alkylamino and oxo groups are substituted.

6. The compound according to any one of claims 1 to 5, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein the general formula (I) has the structure of the following general formula (Ia), in, L, Ring A, R 3 、R 4 、R 5 and p have the definitions given in claims 1-5.

7. The compound according to claim 1 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Selected from 8. The compound according to any one of claims 1 to 7, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein the compound is selected from the following:

9. A pharmaceutical composition comprising the compound according to any one of claims 1 to 8 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and a pharmaceutically acceptable carrier.

10. Use of the compound according to any one of claims 1 to 8 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, or the pharmaceutical composition according to claim 9 in the preparation of a medicament for treating a Cbl-b-mediated disease.

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