Compounds as keap1 activators and uses thereof

Abstract

The present application belongs to the field of medicinal chemistry, and relates to a kind of compound as KEAP1 activator and application thereof, specifically, the present application provides the compound shown in formula (I) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, their preparation method and the pharmaceutical composition containing these compounds and the purposes of these compounds or compositions for treating KEAP1 mediated diseases.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical chemistry, specifically relating to compounds or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs that are KEAP1 activators, methods for their preparation, pharmaceutical compositions containing these compounds, and the use of these compounds or compositions for treating KEAP1-mediated diseases. Background Technology

[0002] In many tumors, constitutive overactivation of the NRF2 protein leads to resistance to chemotherapy and radiotherapy. Blocking NRF2 activity in tumors has been shown to reduce drug resistance in disease models. Studies have shown that under oxidative stress, the cysteine ​​residue at C151 of the KEAP1 protein is covalently modified, reducing the KEAP1-CUL3 protein interaction, which in turn weakens the ubiquitination and degradation of intracellular NRF2 protein. The homeostatic NF2 protein translocates into the nucleus, thereby activating the expression of downstream antioxidant stress-related proteins, leading to enhanced tumor resistance to radiotherapy and chemotherapy.

[0003] Therefore, developing activators of the KEAP1 protein and utilizing the KEAP1-CUL3 E3 ligase system to degrade NRF2, an important protein in tumor cell survival, thereby inhibiting tumor growth and its resistance to radiotherapy and chemotherapy, is a promising therapeutic strategy. Summary of the Invention

[0004] One object of the present invention is to provide a class of compounds, isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs having KEAP1 activating activity as shown in general formulas (I), (Ia), (Ib), (II), and (IIa).

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

[0006] Another object of the present invention is to provide compositions comprising compounds of general formulas (I), (Ia), (Ib), (II), and (IIa) of the present invention, or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, and pharmaceutically acceptable carriers, as well as compositions comprising compounds of general formulas (I), (Ia), (Ib), (II), and (IIa) of the present invention, or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, and one or more other pharmaceutical products.

[0007] Another object of the present invention is to provide a method for treating KEAP1-mediated diseases using compounds of general formulas (I), (Ia), (Ib), (II), and (IIa), or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs of the present invention, and the use of compounds of general formulas (I), (Ia), (Ib), (II), and (IIa), or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs of the present invention in the preparation of medicaments for treating KEAP1-mediated diseases.

[0008] To achieve the aforementioned objectives, the present invention provides the following technical solution:

[0009] In a first aspect, the present invention provides compounds of general formula (I) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs.

[0010]

[0011] in,

[0012] X is selected from O, CH2, and NH;

[0013] L is selected from single bond, -C 1-6 Alkyl groups, -NH groups, -O groups, and -S groups;

[0014] Ring A is selected from aryl, heteroaryl, cycloalkyl, and heterocyclic groups;

[0015] R 1 R 2 R 3 Each is independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino;

[0016] R 4 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C1-6 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl group, oxo group, double C 1-6 Alkylamino, aryl, heteroaryl, cycloalkyl, and heterocyclic groups; and

[0017] m, n, and q are each independently selected from 0, 1, 2, 3, and 4.

[0018] In some embodiments, the compounds of the present invention are compounds of general formula (I) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, wherein general formula (I) has the structure of the following general formula (Ia):

[0019]

[0020] Among them, X, L, ring A, and R 1 R 2 R 3 R 4 , m, n and q have the definitions described in general formula (I).

[0021] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia), or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, wherein L is selected from single bonds, -NH-, and -O-. In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia), or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, wherein ring A is selected from C 6-15 Aryl, 5-15 quinone heteroaryl, C 3-15 Cycloalkyl groups and 3-15 membered heterocyclic groups, wherein the heteroatoms are selected from N, O, S, P and B.

[0022] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 4 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C1-6 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl group, oxo group, double C 1-6 Alkylamino, C 6-15 Aryl, 5-15 heteroaryl, C 3-15 Cycloalkyl and 3-15 membered heterocyclic groups.

[0023] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein ring A is selected from pyrimidinyl groups. , , , , , , , and R 4 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino, q is selected from 0, 1, 2, 3 and 4.

[0024] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein Selected from , , , , , , , , , , and .

[0025] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 1 Selected from C 1-6 Alkyl, C 2-4 alkenyl and C 2-4 Alkyne group.

[0026] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 1 Selected from vinyl.

[0027] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 3 Selected from halogens, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-6 Alkyl, C 1-6 Alkyl acyl and C 1-6 Alkyl sulfonyl group.

[0028] In some specific embodiments, the compounds of the present invention are compounds of general formula (I), general formula (Ia) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 3 Selected from fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, ethynyl, propynyl, trifluoromethyl, and methanesulfonyl.

[0029] In some specific embodiments, the compounds of the present invention are compounds of general formula (I) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, wherein general formula (I) has the structure of the following general formula (Ib):

[0030]

[0031] in,

[0032] Ring B and ring C are each independently selected from C. 6-12 Aryl, 5-8 quinone heteroaryl, C 3-8 Cycloalkyl and 3-8 membered heterocyclic groups, optionally surrounded by one or more groups selected from halogen, hydroxyl, C 1-6 Alkyl, C2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 The alkylamino group is substituted, wherein the heteroatom is selected from N, O, S and P;

[0033] Rings B and C can be either parallel rings or spiral rings;

[0034] Y1 and Y2 are each independently selected from CH and N;

[0035] X, L, R 1 R 2 R 3 , m and n have the definitions described in the general formula (I) above.

[0036] In some embodiments, the compound of general formula (Ib) according to the invention, or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, or a prodrug, wherein the group Selected from , , , , , , , , and .

[0037] This invention provides compounds of general formula (II) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs.

[0038]

[0039] in,

[0040] Ring B and ring C are each independently selected from C. 6-15 Aryl, 5-15 quinone heteroaryl, C 3-15 Cycloalkyl and 3-15 membered heterocyclic groups, optionally surrounded by one or more groups selected from halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino group substitution;

[0041] R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino;

[0042] Y is selected from O, CH2, and NH; and

[0043] r and s are each independently selected from 0, 1, 2, 3 and 4.

[0044] In some embodiments, the compounds of the present invention are compounds of general formula (II) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs, wherein general formula (II) has the structure of general formula (IIa):

[0045]

[0046] Among them, Y, ring B, ring C, and R 5 R 6 R 7 r and s have the definitions described in general formula (II).

[0047] In some specific embodiments, the present invention comprises compounds of general formula (II) or general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvent compounds, crystals or prodrugs, wherein ring B is selected from C. 6-15 aryl, 5-15-membered heteroaryl, optionally substituted with one or more elements selected from halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 The alkylamino group is substituted, and the heteroatom is selected from N, O, P, S and B.

[0048] In some specific embodiments, the present invention comprises compounds of general formula (II), general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvent compounds, crystals or prodrugs, wherein ring B is selected from pyrimidines, optionally by one or more elements selected from halogens, hydroxyl groups, C... 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 The alkylamino group is substituted; further, it is optionally replaced by one or more groups selected from fluorine, chlorine, bromine, iodine, hydroxyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, C 1-3 Alkoxy, halogenated C 1-3 Group substitutions of alkoxy, cyano, amino, and oxo groups.

[0049] In some specific embodiments, the present invention comprises compounds of general formula (II) or general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvent compounds, crystals or prodrugs, wherein the ring C is selected from C 6-15 aryl, 5-15-membered heteroaryl, and 3-15-membered heterocyclic groups, optionally composed of one or more elements selected from halogen, hydroxyl, C... 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 The alkylamino group is substituted, and the heteroatom is selected from N, O, P, S and B.

[0050] In some specific embodiments, the present invention comprises compounds of general formula (II), general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvent compounds, crystals or prodrugs, wherein ring C is selected from pyrazole, pyridine, , and Its optional component is selected from one or more elements chosen from halogens, hydroxyl groups, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 The alkylamino group is substituted; further, it is optionally substituted with one or more groups selected from fluorine, chlorine, bromine, iodine, hydroxyl, methyl, ethyl, propyl, isopropyl, trifluoromethyl, C 1-3 Alkoxy, halogenated C 1-3 Group substitutions of alkoxy, cyano, amino, and oxo groups.

[0051] In some specific embodiments, the compounds of the present invention are compounds of general formula (II), general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 5 Selected from C 1-6 Alkyl, C 2-4 alkenyl and C 2-4 Alkyne group.

[0052] In some specific embodiments, the compounds of the present invention are compounds of general formula (II), general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 5 Selected from vinyl.

[0053] In some specific embodiments, the compounds of the present invention are compounds of general formula (II), general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 7 Selected from halogens, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, halogenated C 1-6 Alkyl, C 1-6 Alkyl acyl and C 1-6 Alkyl sulfonyl group.

[0054] In some specific embodiments, the compounds of the present invention are compounds of general formula (II), general formula (IIa) or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, wherein R 7 Selected from fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, ethynyl, propynyl, trifluoromethyl, and methanesulfonyl.

[0055] This invention provides the following specific compounds or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs:

[0056] , , ,

[0057] , , ,

[0058] , , ,

[0059] , , ,

[0060] , , ,

[0061] , , ,

[0062] , , and

[0063] .

[0064] In a second aspect, the present invention provides pharmaceutical compositions comprising the compounds of the present invention or isomers thereof, pharmaceutically acceptable salts, solvates, crystals, or prodrugs.

[0065] In some embodiments, the present invention provides compounds or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs of the present invention, and pharmaceutical compositions comprising compounds or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs of the present invention for treating KEAP1-mediated diseases.

[0066] In some embodiments, the present invention provides pharmaceutical compositions comprising the compound of the present invention or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal or prodrug, and a pharmaceutically acceptable carrier.

[0067] The compounds of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals, or prodrugs can be mixed with pharmaceutically acceptable carriers, diluents, or excipients to prepare pharmaceutical formulations 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 formulations can be administered via any route, such as by infusion or bolus, or by absorption through the epithelium or mucous membranes of the skin (e.g., oral mucosa or rectum). Administration can be systemic or local. Examples of oral formulations include solid or liquid dosage forms, specifically including tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The formulations can be prepared by methods known in the art and contain carriers, diluents, or excipients conventionally used in the field of pharmaceutical formulations.

[0068] Thirdly, the present invention provides a method for treating KEAP1-mediated diseases using compounds or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, or pharmaceutical compositions comprising thereof, represented by formulas (I), (Ia), (Ib), (II), and (IIa), as well as their use in the preparation of medicaments for treating KEAP1-mediated diseases.

[0069] In some preferred embodiments, the present invention provides methods for treating KEAP1-mediated diseases using compounds or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, or pharmaceutical compositions comprising thereof, as shown in formulas (I), (Ia), (Ib), (II), and (IIa), and their uses in the preparation of medicaments for treating KEAP1-mediated diseases, wherein the KEAP1-mediated diseases include, but are not limited to, proliferative diseases, metabolic diseases, or hematologic disorders. In some embodiments, the KEAP1-mediated diseases described in the present invention are cancer.

[0070] In some embodiments, the KEAP1-mediated diseases described in this invention include, but are not limited to, acoustic neuroma, adenocarcinoma, adrenal carcinoma, anal cancer, angiosarcoma (e.g., lymphangiosarcoma, lymphangioendothelial sarcoma, angiosarcoma), adnexal cancer, benign monoclonal gammopathy, biliary cancer (e.g., cholangiocarcinoma), bladder cancer, breast cancer (e.g., breast adenocarcinoma, papillary breast carcinoma, breast cancer, medullary breast carcinoma, triple-negative breast cancer), brain cancer (e.g., meningioma; glioma, e.g., astrocytoma, oligodendroglioma; medulloblastoma), bronchial cancer, carcinoid tumor, cervical cancer (e.g., cervical adenocarcinoma), choriocarcinoma, craniopharyngioma, colorectal cancer (e.g., colon cancer, rectal cancer, colorectal adenocarcinoma), epithelial cancer, ependymoma, endothelial sarcoma (e.g., Kaposi's sarcoma). Sarcoma, multiple idiopathic hemorrhagic sarcomas, endometrial cancer (e.g., uterine cancer, uterine sarcoma), esophageal cancer (e.g., esophageal adenocarcinoma, Barrett's adenocarcinoma), Ewing sarcoma Cancers include sarcoma, ocular cancer (e.g., intraocular melanoma, retinoblastoma), familial eosinophilia, gallbladder cancer, gastric 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., laryngeal cancer, pharyngeal cancer, nasopharyngeal cancer, oropharyngeal cancer)), hematopoietic system cancers (e.g., leukemia 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 CLL, 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, etc. Burkitt lymphoma, lymphoplasmacytic lymphoma (i.e., Waldenström's macroglobulinemia), hairy cell leukemia (HCL), immunoblastic large cell lymphoma, precursor B-cell lymphoblastic lymphoma, and primary central nervous system (CNS) lymphoma; and T-cell NHL such as precursor T-cell 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); or a mixture of one or more leukemias / lymphomas as described above.And multiple myeloma (MM), heavy chain diseases (e.g., alpha chain disease, gamma chain disease, μ chain disease), angioblastoma, inflammatory myofibroblastoma, immune cell amyloidosis, renal cell carcinoma (e.g., nephroblastoma, also known as Wilms' tumor), hepatocellular carcinoma (e.g., hepatocellular carcinoma (HCC), malignant hepatocellular carcinoma), lung cancer (e.g., bronchial carcinoma, small cell lung cancer (SCLC), non-small cell lung cancer (NSCLC), lung adenocarcinoma), leiomyosarcoma (LMS), mastocytosis (e.g., generalized mastocytosis), myelodyplastic syndrome (MDS), mesothelioma, myeloproliferative disorders (MPD) (e.g., polycythemia vera (PV), idiopathic thrombocythemia (ET), idiopathic extramedullary metaplasia (AMM)). This includes myelofibrosis (MF), chronic idiopathic myelofibrosis, chronic myeloid leukemia (CML), chronic neutrophilic leukemia (CNL), eosinophilic leukocytosis (HES), neuroblastoma, neurofibroma (e.g., type 1 or 2 multiple neurofibroma (NF), Schwannoma), neuroendocrine carcinoma (e.g., gastrointestinal pancreatic 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 myxoma (IPMN), islet cell tumor), and penile cancer (e.g., Paget's disease of the penis and scrotum). Diseases including pineal gland tumors, primary neuroectodermal tumors (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 bowel cancer (e.g., adnexal cancer), soft tissue sarcomas (e.g., malignant fibrous histiocytoma (MFH), liposarcoma, malignant peripheral nerve sheath tumor (MPNST), chondrosarcoma, fibrosarcoma, myxosarcoma), sebaceous gland cancer, sweat gland cancer, synovial tumors, testicular cancer (e.g., seminoma, embryonal testicular carcinoma), thyroid cancer (e.g., papillary thyroid carcinoma, papillary thyroid carcinoma (PTC), medullary thyroid carcinoma), urethral cancer, vaginal cancer, and vulvar cancer (e.g., vulvar Paget's disease), medulloblastoma, adenoid cystic carcinoma, melanoma, and glioblastoma.

[0071] In some preferred embodiments, the present invention provides a method for treating KEAP1-mediated diseases using compounds or isomers thereof, pharmaceutically acceptable salts, solvates, crystals or prodrugs, or pharmaceutical compositions comprising thereof, represented by general formulas (I), (Ia), (Ib), (II), and (IIa), as well as their use in the preparation of medicaments for treating KEAP1-mediated diseases, wherein the KEAP1-mediated diseases include, but are not limited to, breast cancer, esophageal cancer, bladder cancer, and lung cancer. Hematopoietic system cancers, 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, hemoglobinopathies, diabetes, and obesity.

[0072] Terminology Definition

[0073] Unless otherwise stated, the terms used in the specification and claims have the following meanings.

[0074] In the compounds of this invention, "hydrogen," "carbon," and "oxygen" include all their isotopes. Isotopes should be understood to include those atoms having the same number of atoms 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.

[0075] In this invention, "isomers" refers to molecules with the same atomic composition and bonding but different three-dimensional spatial 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 are capable of rotating the plane of polarized light. In describing optically active compounds, the prefixes D, L, or R, S are used to indicate the absolute configuration of the chiral center of the molecule. The prefixes D, L, or (+), (-) are used to name the symbols for the plane polarization rotation of the compound; (-) or L indicates that the compound is levorotatory, and the prefix (+) or D indicates that the compound is dextrorotatory. These stereoisomers have the same chemical structure but different stereostructures. Specific stereoisomers can be enantiomers, and mixtures of isomers are usually called enantiomer mixtures. A 50:50 enantiomer mixture is called a racemic mixture or racemate, which may result in a lack of stereoselectivity or stereodirection during chemical reactions. The terms “racemic mixture” and “racemate” refer to a mixture of two equimolar enantiomers that lack optical activity.

[0076] Depending on the choice of starting materials and methods, the compounds of this invention can exist as one or a mixture of possible isomers, such as racemic mixtures and mixtures of non-corresponding isomers (depending on the number of asymmetric carbon atoms). Optically active (R)- or (S)- isomers can be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

[0077] Any mixture of stereoisomers obtained can be separated into pure or substantially pure geometric isomers, enantiomers, and diastereomers based on differences in the physicochemical properties of the components, for example, by chromatography and / or fractional crystallization.

[0078] In this invention, "halogen" refers to fluorine, chlorine, bromine, or iodine. "Halogenated" in this invention means substituted with fluorine, chlorine, bromine, or iodine.

[0079] In this invention, "alkyl" refers to a straight-chain or branched saturated aliphatic hydrocarbon group, preferably a straight-chain or branched group containing 1 to 6 carbon atoms, and more preferably a straight-chain or branched group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc. The alkyl group can be substituted or unsubstituted, and when substituted, the substituent can be at any usable connection point.

[0080] In this invention, "carbonyl" and "acyl" both refer to -C(O)-.

[0081] In this invention, "sulfonyl" refers to -S(O)2-.

[0082] In this invention, "sulfonamide group" refers to -S(O)2NH-.

[0083] In this invention, "halogenated alkyl" refers to an alkyl group that is substituted with at least one halogen.

[0084] In this invention, "hydroxyalkyl" refers to an alkyl group that is substituted with at least one hydroxyl group.

[0085] In this invention, "alkoxy" refers to -O-alkyl. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy, etc. Alkoxy groups can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any usable connection point.

[0086] In this invention, "cycloalkyl" refers to a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups can be substituted or unsubstituted monocyclic groups having 3-15 carbon atoms, or fused, bridged, or spirocyclic bicyclic or tricyclic saturated hydrocarbon groups, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0087] The term "heterocyclic group" in this invention refers to a group ("3-15-membered heterocyclic group") having 1 to 4 cyclic heteroatoms (each heteroatom independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon) in a 3- to 15-membered non-aromatic ring system. In a heterocyclic group containing one or more nitrogen atoms, the linking point can be a carbon or nitrogen atom, provided the valence permits. The heterocyclic group can be monocyclic ("monocyclic heterocyclic group") or a fused, bridged, or spirocyclic polycyclic system (e.g., a bicyclic system (also called "bicyclic heterocyclic group")), and can be saturated or partially unsaturated. Suitable heterocyclic groups include, but are not limited to, piperidinyl, aziridine, aziridine propane, tetrahydropyrroleyl, piperazineyl, dihydroquinazolinyl, oxacyclopropyl, oxacyclobutyl, tetrahydrofuranyl, tetrahydropyranyl, 6-aziridine[3.4]octyl, , Each instance of a heterocyclic group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any usable connection point.

[0088] In this invention, "aryl" refers to an aromatic system that may comprise a monocyclic or fused polycyclic ring, preferably a monocyclic or fused bicyclic aromatic system, containing 6 to 15 carbon atoms, more preferably about 6 to about 10 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthraceneyl, fluorenyl, and indanyl. The aryl group may be optionally substituted or unsubstituted, and when substituted, the substituent may be at any usable connection point.

[0089] In this invention, "heteroaryl" refers to an aryl group in which at least one carbon atom is replaced by a heteroatom. The heteroaryl group may have a monocyclic or fused, bridged, or spirocyclic polycyclic system, wherein at least one ring is aromatic. Preferably, it consists of 5-15 atoms (5-15-membered heteroaryl), more preferably 5-10 atoms (5-10-membered heteroaryl), wherein the heteroatom is O, S, N, or B. The heteroaryl groups include, but are not limited to, imidazolyl, pyrrolyl, furanyl, thiophene, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, indoleyl, pyridyl, pyrimidinyl, pyrazinyl, triazinyl, isoindoleyl, benzopyrazolyl, benzimidazolyl, benzofuranyl, benzopyranyl, benzothiophene, benzooxazolyl, benzothiazolyl, benzothiazolyl, benzoisooxazolyl, benzoisothiazolyl, quinolinyl, isoquinolinyl, quinazolinyl, cenolinyl, quinoxazinyl, benzothiazolyl, imidazopyridyl, pyrimidinylpyrazolyl, pyrimidinylimidazole, benzo[d]thiazolyl, spiro[ Indoline-3,4'-piperidin]-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]pyridyl, 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]pyridyl, , , , , , , , , The heteroaryl group can be optionally substituted or unsubstituted; when substituted, the substituent can be at any usable connection point.

[0090] The term "pharmaceutically acceptable salt" in this invention refers to salts of the compounds of this invention that are safe and effective when used in mammals and possess the intended biological activity.

[0091] In the conventional sense, the term "solvent" in this invention refers to a complex formed by a combination of a solute (such as an active compound or a salt of an active compound) and a solvent (such as water). The solvent refers to a solvent known or readily identifiable to those skilled in the art. If water is present, the solvate is typically referred to as a hydrate, such as a hemihydrate, monohydrate, dihydrate, trihydrate, or a substitute thereof.

[0092] The in vivo effects of compounds having chemical formulas (I) and (II) can be partially exerted by one or more metabolites formed in the human or animal body after administration of the compounds having chemical formulas (I) and (II). As described above, the in vivo effects of compounds having chemical formulas (I) and (II) can also be exerted via the metabolism of prodrugs ("prodrugs"). The "prodrug" of this invention refers to a compound that, under physiological conditions in an organism, is converted into the compounds of this invention through reaction with enzymes, gastric acid, etc., i.e., a compound converted into the compounds of this invention through enzyme oxidation, reduction, hydrolysis, etc., and / or through hydrolysis reactions such as gastric acid, etc.

[0093] The “crystallization” of this invention refers to a solid whose internal structure is formed by the regular repetition of atoms (or groups thereof) in three dimensions, which is different from amorphous solids that do not have such a regular internal structure.

[0094] The term "pharmaceutical composition" as used in this invention refers to a mixture comprising any of the compounds described herein, including corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts or their chemically protected forms, and one or more pharmaceutically acceptable carriers and / or mixtures of other one or more drugs. The purpose of a pharmaceutical composition is to facilitate the administration of the compound to a living organism. Such compositions are typically used in the preparation of medicaments for the treatment and / or prevention of diseases mediated by one or more kinases.

[0095] The "pharmaceutical-grade carrier" of this invention refers to a carrier that does not cause significant irritation to the organism and does not interfere with the biological activity and properties of the administered compound. This includes all solvents, diluents or other excipients, dispersants, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc., unless any conventional carrier medium is incompatible with the compounds of this invention. Some 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 carboxymethyl cellulose, as well as cellulose and cellulose acetate; malt, gelatin, etc.

[0096] In this invention, "excipient" refers to an inert substance added to a pharmaceutical composition to further promote the delivery of the compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycol. Detailed Implementation

[0097] The present invention will be further described in detail below with reference to the embodiments, but the present invention is not limited to these embodiments. Unless otherwise specified, all materials used in the following embodiments are commercially available.

[0098] Intermediate 1: (R)-1-(3-(3-bromo-5-chlorophenyl)morpholinyl)prop-2-en-1-one

[0099]

[0100] Step 1: Preparation of (R)-1-(3-(3-bromo-5-chlorophenyl)morpholinyl)prop-2-en-1-one

[0101]

[0102] (R)-3-(3-bromo-5-chlorophenyl)morpholine (10 g, 36.15 mmol) was added to a two-necked flask, dissolved in dichloromethane (100 mL), followed by the addition of triethylamine (7.5 mL, 54.23 mmol). The mixture was purged three times with argon gas, and then acryloyl chloride (3 mL, 43.39 mmol) was added. The reaction was allowed to proceed at room temperature for 2 hours. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by column chromatography to give 870 mg of the title compound as a white solid. ESI-MS m / z: 329.8, 331.9 [M+H] +

[0103] Intermediate 2: (R)-1-(3-(3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)phenyl)morpholinyl)prop-2-en-1-one

[0104]

[0105] Step 1: Preparation of (R)-1-(3-(3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxoborhecyclopentan-2-yl)phenyl)morpholinyl)prop-2-en-1-one

[0106]

[0107] (R)-1-(3-(3-bromo-5-chlorophenyl)morpholinyl)prop-2-en-1-one (1 g, 3.02 mmol) was added to a microwave tube, dissolved in 1,4-dioxane (10 mL), followed by the addition of pinacol diboronate (1.115 g, 4.53 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (110 mg, 0.151 mmol), and potassium acetate (741 mg, 2.55 mmol). The mixture was purged three times with argon gas and microwaved at 80 °C for 2 hours. Three portions were added. After the reaction was complete, the mixture was concentrated under reduced pressure and purified by column chromatography to give 2.5 g of the title compound as an orange oil.

[0108] ESI-MS m / z: 378.0 [M+H] +

[0109] Example 1: (R)-1-(3-(3-(4-(1H-pyrazol-5-yl)pyrimidin-2-yl)-5-chlorophenyl)morpholino)prop-2-en-1-one

[0110]

[0111] Step 1: Preparation of 2-chloro-4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)pyrimidine

[0112]

[0113] 4-Bromo-2-chloropyrimidine (285 mg, 1.47 mmol), 1-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)-1H-pyrazole (409 mg, 1.47 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (108 mg, 0.147 mmol), and potassium carbonate (608 mg, 4.41 mmol) were added to a microwave-safe tube. 10 mL of 1,4-dioxane and 2 mL of water were added. The mixture was microwave-safe at 100 °C for 1 hour under argon protection. The reaction was monitored by LC-MS. After the reaction was complete, column chromatography was performed to obtain the title product. ESI-MS m / z: 264.9 [M+H] +

[0114] Step 2: Preparation of (R)-1-(3-(3-chloro-5-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)pyrimidin-2-yl)phenyl)morpholino)prop-2-en-1-one

[0115]

[0116] 2-Chloro-4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)pyrimidine (170 mg, 0.45 mmol), (R)-1-(3-(3-chloro-5-(4,4,5,5-tetramethyl-1,3,2-dioxoboronyl-2-yl)phenyl)morpholinyl)prop-2-en-1-one (119 mg, 0.45 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (33 mg, 0.045 mmol), and potassium carbonate (124 mg, 0.9 mmol) were added to a microwave tube, along with 5 mL of 1,4-dioxane and 1 mL of water. The mixture was microwaved at 100 °C for 2 hours under argon protection. The reaction was monitored by LCMS. After the reaction was complete, column chromatography was performed to obtain the title product. ESI-MS m / z: 480.0 [M+H] +

[0117] Step 3: Preparation of (R)-1-(3-(3-(4-(1H-pyrazol-5-yl)pyrimidin-2-yl)-5-chlorophenyl)morpholino)prop-2-en-1-one

[0118]

[0119] (R)-1-(3-(3-chloro-5-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)pyrimidin-2-yl)phenyl)morpholino)prop-2-en-1-one (100 mg, 0.21 mmol) was dissolved in dichloromethane (5 mL) in a single-necked flask, and trifluoroacetic acid (2 mL) was added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete, the solution was concentrated under reduced pressure and purified by C18 column chromatography to give 49 mg of the title product as a white solid. ESI-MS m / z: 395.9 [M+H] + . 1 H NMR (400 MHz, d6-DMSO) δ 13.45(s, 1H), 8.92 (s, 1H), 8.63 – 8.26 (m, 2H), 8.13 – 7.77 (m, 2H), 7.61 (s,1H), 7.25 – 7.02 (m, 1H), 7.02 – 6.79 (m, 1H), 6.39 – 6.17 (m, 1H), 5.94 –5.72 (m, 1H), 5.71 – 5.36 (m, 1H), 4.63 – 4.43 (m, 1H), 4.19 – 3.68 (m, 3H),3.62 – 3.46 (m, 1H), 3.11 – 2.79 (m, 1H).

[0120] Example 2: (R)-1-(3-(3-(4-(1H-pyrazol-4-yl)pyrimidin-2-yl)-5-chlorophenyl)morpholino)prop-2-en-1-one

[0121]

[0122] Step 1: Preparation of 2-chloro-4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)pyrimidine

[0123]

[0124] 2,4-Dichloropyrimidine (603 mg, 4.05 mmol), 1-(tetrahydro-2H-pyran-2-yl)-4-(4,4,5,5-tetramethyl1,3,2-dioxoboronyl-2-yl)-1H-pyrazole (750 mg, 2.7 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (296 mg, 0.405 mmol), and potassium phosphate (2.6 g, 12.15 mmol) were added to a microwave-safe tube. 15 mL of 1,4-dioxane and 3 mL of water were added. The mixture was microwave-safe at 100 °C for 1 hour under argon protection. The reaction was monitored by LC-MS. After the reaction was complete, column chromatography was performed to obtain the title product. ESI-MS m / z: 265.0 [M+H] +

[0125] Step 2: Preparation of 1-((3R)-3-(3-chloro-5-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)pyrimidin-2-yl)phenyl)morpholino)prop-2-en-1-one

[0126]

[0127] 2-Chloro-4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)pyrimidine (119 mg, 0.45 mmol), intermediate 2 (170 mg, 0.45 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (33 mg, 0.045 mmol), and potassium carbonate (125 mg, 0.90 mmol) were added to a microwave-safe tube. 10 mL of 1,4-dioxane and 2 mL of water were added. The mixture was reacted under argon protection at 100 °C for 2 hours. The reaction was monitored by LC-MS. After the reaction was complete, column chromatography was performed to obtain the title product. ESI-MS m / z: 480.0 [M+H] +

[0128] Step 3: Preparation of (R)-1-(3-(3-(4-(1H-pyrazol-4-yl)pyrimidin-2-yl)-5-chlorophenyl)morpholino)prop-2-en-1-one

[0129]

[0130] 1-((3R)-3-(3-chloro-5-(4-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-4-yl)pyrimidin-2-yl)phenyl)morpholino)prop-2-en-1-one (161 mg, 0.34 mmol) was dissolved in dichloromethane (5 mL) in a single-necked flask, and trifluoroacetic acid (2 mL) was added. The mixture was stirred at room temperature for 2 hours. After the reaction was complete as monitored by LC-MS, the mixture was concentrated under reduced pressure. 18 Purification by column chromatography yielded 62 mg of the title product as a white solid. ESI-MS m / z: 396.0 [M+H] + . 1 H NMR (400 MHz, d6-DMSO) δ 13.42 (s, 1H), 8.82 (d, J = 5.3 Hz, 1H), 8.66 (s, 1H), 8.41 (s, 1H),8.46 – 8.36 (m, 2H), 7.74 (d, J = 5.3 Hz, 1H), 7.59 (s, 1H), 7.01 – 6.84 (m,1H), 6.35 – 6.25 (m, 1H), 5.81 (d, J = 10.0 Hz, 1H), 5.70 – 5.37 (m, 1H), 4.57 – 4.48 (m, 1H), 4.13 – 3.74 (m, 3H), 3.61 – 3.48 (m, 1H), 3.31 – 3.12 (m, 1H).

[0131] Example 3: (R)-1-(3-(3-chloro-5-(4-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-yl)phenyl)morpholinyl)prop-2-en-1-one

[0132]

[0133] Step 1: Preparation of 2-chloro-4-(1-methyl-1H-pyrazol-4-yl)pyrimidine

[0134]

[0135] To a 50 mL round-bottom flask, add 2-chloro-4-bromopyrimidine (290 mg, 1.5 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxoborhexacyclopentan-2-yl)-1H-pyrazole (343 mg, 1.65 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (109 mg, 1.65 mmol), potassium phosphate (955 mg, 4.5 mmol), 15 mL of 1,4-dioxane, and 3 mL of water. Purge the gas three times with argon, and heat at 90 °C with stirring for 2.5 hours. After the reaction was complete as detected by LC-MS, cool to room temperature, quench the reaction with water, and extract by liquid-liquid extraction. Extract the aqueous phase three times with ethyl acetate, combine the organic phases, wash twice with saturated brine, dry to anhydrous sodium sulfate, concentrate under reduced pressure, and column chromatography to obtain 230 mg of the title product as a pale yellow solid. ESI-MS m / z: 195.0 [M+H] +

[0136] Step 2: Preparation of (R)-1-(3-(3-chloro-5-(4-(1-methyl-1H-pyrazol-4-yl)pyrimidin-2-yl)phenyl)morpholino)prop-2-en-1-one

[0137]

[0138] Add 2-chloro-4-(1-methyl-1H-pyrazol-4-yl)pyrimidine (86 mg, 0.4 mmol), intermediate 2 (152 mg, 0.44 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (29 mg, 0.04 mmol), potassium phosphate (255 mg, 1.2 mmol), 4 mL of 1,4-dioxane, and 0.8 mL of water to a 25 mL round-bottom flask. Purge the mixture three times with argon gas, heat at 90 °C with stirring overnight. After LC-MS analysis, cool to room temperature, quench the reaction with water, and extract by liquid-liquid extraction. Extract the aqueous phase three times with ethyl acetate, combine the organic phases, wash twice with saturated brine, dry to anhydrous sodium sulfate, concentrate under reduced pressure, and then... 18 Column chromatography purification yielded 80 mg of the title product. ESI-MS m / z: 409.9 [M+H] + . 1H NMR (400 MHz, d6-DMSO) δ 8.81 (d, J = 5.3 Hz, 1H), 8.60 (s, 1H), 8.51 – 8.27 (m, 2H), 8.25 (s,1H), 7.68 (d, J = 5.3 Hz, 1H), 7.58 (s, 1H), 7.01 – 6.81 (m, 1H), 6.35 – 6.18(m, 1H), 5.91 – 5.75 (m, 1H), 5.70 – 5.34 (m, 1H), 4.58 – 4.44 (m, 1H), 3.95(s, 3H), 3.94 – 3.69 (m, 3H), 3.60 – 3.43 (m, 1H), 3.27 – 2.81 (m, 1H).

[0139] Example 4: (R)-4-(2-(3-(4-acryloylmorpholin-3-yl)-5-chlorophenyl)pyrimidin-4-yl)pyridin-2(1H)-one

[0140]

[0141] Step 1: Preparation of 2-chloro-4-(2-methoxypyridin-4-yl)pyrimidine

[0142]

[0143] 4-Bromo-2-chloropyrimidine (300 mg, 1.26 mmol), (2-methoxypyridin-4-yl)boronic acid (193 mg, 1.26 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (92 mg, 0.126 mmol), and potassium carbonate (348 mg, 2.52 mmol) were added to a microwave-safe tube. 5 mL of 1,4-dioxane and 1 mL of water were added. The mixture was microwave-safe at 80 °C for 1 hour under argon protection. The reaction was monitored by LC-MS. After the reaction was complete, column chromatography was performed to obtain the title product. ESI-MS m / z: 222.0 [M+H] +

[0144] Step 2: Preparation of 4-(2-chloropyrimidin-4-yl)pyridin-2-ol

[0145]

[0146] 2-Chloro-4-(2-methoxypyridin-4-yl)pyrimidine (230 mg, 1.04 mmol) was dissolved in N,N'-dimethylformamide (5 mL), followed by the addition of p-toluenesulfonic acid (895 mg, 5.2 mmol) and lithium chloride (220 mg, 5.2 mmol). The mixture was stirred at 110 °C for 3 hours. After the reaction was completed, water was added to quench the reaction, resulting in the precipitation of a green solid. The solid was filtered and dried to give 120 mg of the title product. ESI-MS m / z: 208.0 [M+H] +

[0147] Step 3: Preparation of (R)-4-(2-(3-(4-acryloylmorpholin-3-yl)-5-chlorophenyl)pyrimidin-4-yl)pyridin-2(1H)-one

[0148]

[0149] 4-(2-chloropyrimidin-4-yl)pyridin-2-ol (70 mg, 0.34 mmol), intermediate 2 (128 mg, 0.34 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (25 mg, 0.034 mmol), and potassium carbonate (94 mg, 0.68 mmol) were added to a microwave-safe tube, along with 5 mL of 1,4-dioxane and 1 mL of water. The mixture was reacted under argon protection at 100 °C for 2 hours. The reaction was monitored by LC-MS. After the reaction was complete, column chromatography was performed to obtain the title product. ESI-MS m / z: 422.9 [M+H] + . 1 HNMR (400 MHz, d6-DMSO) δ 11.90 (s, 1H), 9.07 (d, J = 5.2 Hz, 1H), 8.42 (s,1H), 8.38 (s, 1H), 8.11 (d, J = 5.3 Hz, 1H), 7.63 (s, 1H), 7.59 (d, J = 6.8Hz, 1H), 7.27 (d, J = 1.1 Hz, 1H), 7.05 – 6.99 (m, 1H), 6.97 – 6.87 (m, 1H), 6.35 – 6.24 (m, 1H), 5.86 – 5.78 (m, 1H), 5.72 – 5.40 (m, 1H), 4.56 – 4.45(m, 1H), 4.13 – 3.72 (m, 3H), 3.61 – 3.49 (m, 1H), 3.29 – 3.10 (m, 1H).

[0150] Example 5: (R)-1-(3-(3-((4-aminopyrimidin-2-yl)amino)-5-chlorophenyl)morpholino)prop-2-en-1-one

[0151]

[0152] Step 1: Preparation of (R)-(3-(4-acryloylmorpholin-3-yl)-5-chlorophenyl)carbamate tert-butyl ester

[0153]

[0154] Intermediate 1 (600 mg, 1.814 mmol) and tert-butyl carbamate (277 mg, 2.359 mmol) were added to a microwave-safe tube, dissolved in 1,4-dioxane (10 mL). Then, 2-(dicyclohexylphosphine)-3,6-dimethoxy-2'-4'-6'-tris-1-propyl-11'-biphenyl (195 mg, 0.362 mmol), tris(dibenzylacetone)dipalladium (166 mg, 0.181 mmol), and cesium carbonate (1.774 g, 5.444 mmol) were added. The mixture was purged three times with argon, heated to 100 °C, and microwaved for 2 hours. After the reaction, the mixture was concentrated under reduced pressure and purified by column chromatography to give 300 mg of the yellow oil title product. ESI-MS m / z: 366.9 [M+H] +

[0155] Step 2: Preparation of (R)-1-(3-(3-amino-5-chlorophenyl)morpholino)prop-2-en-1-one

[0156]

[0157] (R)-(3-(4-acryloylmorpholin-3-yl)-5-chlorophenyl)carbamate tert-butyl ester (300 mg, 0.817 mmol) was added to a single-necked flask, dissolved in dichloromethane (10 mL), followed by trifluoroacetic acid (2 mL). The reaction was carried out at room temperature for 2 hours. After the reaction was complete, the mixture was concentrated under reduced pressure, diluted with ethyl acetate, washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 175 mg of the title product, a brown oil. ESI-MS m / z: 266.9 [M+H] +

[0158] Step 3: Preparation of (R)-(2-((3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)amino)pyrimidin-4-yl)(tert-butoxycarbonyl)carbamate tert-butyl ester

[0159]

[0160] (R)-1-(3-(3-amino-5-chlorophenyl)morpholino)prop-2-en-1-one (172 mg, 0.644 mmol) and (2-bromopyrimidin-4-yl)(tert-butyloxycarbonyl)carbamate tert-butyl ester (362 mg, 0.967 mmol) were added to a microwave-safe tube. 1,4-Dioxane (10 mL) was added to dissolve the tert-butyl carbamate. Then, 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (75 mg, 0.128 mmol), palladium acetate (14 mg, 0.064 mmol), and cesium carbonate (630 mg, 1.934 mmol) were added. The mixture was purged three times with argon, heated to 100 °C, and microwaved for 2 hours. After the reaction, the mixture was concentrated under reduced pressure and purified by column chromatography to give 227 mg of the yellow oil title product. ESI-MS m / z: 560.0 [M+H] +

[0161] Step 4: Preparation of (R)-1-(3-(3-((4-aminopyrimidin-2-yl)amino)-5-chlorophenyl)morpholino)prop-2-en-1-one

[0162]

[0163] (R)-(2-((3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)amino)pyrimidin-4-yl)(tert-butoxycarbonyl)carbamate tert-butyl ester (190 mg, 0.654 mmol) was added to a single-necked flask, dissolved in dichloromethane (10 mL), followed by trifluoroacetic acid (3 mL). The reaction was carried out at room temperature for 2 hours. After the reaction was completed, the mixture was concentrated under reduced pressure, diluted with ethyl acetate, washed with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, purified by C18 column chromatography, and lyophilized to give 46 mg of the title product as a white powder. ESI-MS m / z: 360.0 [M+H] + . 1 H NMR (400 MHz, d6-DMSO) δ 9.19 (s, 1H), 7.99– 7.62 (m, 3H), 6.93 – 6.76 (m, 2H), 6.59 (s, 2H), 6.27 – 6.16 (m, 1H), 5.94(d, J = 5.7 Hz, 1H), 5.74 (d, J = 9.3 Hz, 1H), 5.49 – 5.17 (m, 1H), 4.43 –4.14 (m, 2H), 4.04 – 3.65 (m, 3H), 3.52 – 3.42 (m, 1H).

[0164] Example 6: (R)-7'-(3-(4-acryloylmorpholin-3-yl)-5-chlorophenyl)-2'3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

[0165]

[0166] Step 1: Preparation of methyl 5-bromo-2-(cyanomethyl)benzoate

[0167]

[0168] 5-Bromo-2-(bromomethyl)benzoate (5 g, 16.24 mmol), trimethylcyanosilane (2.44 mL, 19.49 mmol), potassium carbonate (4.49 g, 32.48 mmol), and cesium fluoride (4.94 g, 32.48 mmol) were dissolved in acetonitrile (30 mL) in a three-necked flask. After purging with argon, the mixture was refluxed at 80 °C. After the reaction was complete as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and column chromatography yielded 2.272 g of the title product. ESI-MS m / z: 253.8, 256.0 [M+H] +

[0169] Step 2: Preparation of methyl 5-bromo-2-(1-cyanocyclopropyl)benzoate

[0170]

[0171] 2.70 g (10.63 mmol) of methyl 5-bromo-2-(cyanomethyl)benzoate was dissolved in dimethyl sulfoxide (20 mL) in a three-necked flask. After purging with argon, sodium hydride (850 mg, 21.26 mmol) was added at 0 °C. After stirring for 1 hour, 1,2-dibromoethane was added, and the mixture was transferred to room temperature and reacted for 2 hours. After the reaction was complete as monitored by LC-MS, the reaction solution was quenched with saturated ammonium chloride solution at 0 °C. The solution was then extracted three times with ethyl acetate. The organic layer was collected, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give 1.532 g of the title product. ESI-MS m / z: 279.9, 281.9 [M+H] +

[0172] Step 3: Preparation of 7'-bromo-2'3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

[0173]

[0174] Methyl 5-bromo-2-(1-cyanocyclopropyl)benzoate (1.53 g, 5.47 mmol) and cobalt dichloride (1.42 g, 10.94 mmol) were dissolved in methanol (15 mL) in a three-necked flask. After purging with argon, sodium borohydride (1.03 g, 27.35 mmol) was added in portions at 0 °C, and the mixture was then transferred to room temperature for reaction. After the reaction was complete as monitored by LC-MS, the reaction solution was concentrated under reduced pressure, and 527 mg of the title product was obtained by column chromatography.

[0175] ESI-MS m / z: 251.9, 253.9 [M+H] +

[0176] Step 4: Preparation of (R)-7'-(3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)-2'3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one

[0177]

[0178] 7'-bromo-2'3'-dihydro-1'H-spiro[cyclopropane-1,4'-isoquinoline]-1'-one (101 mg, 0.40 mmol), intermediate 2 (150 mg, 0.40 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium(II) dichloride (29 mg, 0.04 mmol), and potassium carbonate (166 mg, 1.20 mmol) were dissolved in a 4 mL mixture of dioxane and water (5:1). After purging with argon, the mixture was reacted at 100 °C for 2 hours. After the reaction was complete as monitored by LC-MS, the reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain 66 mg of the title product. ESI-MS m / z: 422.9 [M+H] + . 1H NMR (400 MHz, d6-DMSO) δ8.17 (s, 1H), 8.09 (d, J = 2.0 Hz, 1H), 7.79 – 7.72 (m, 1H), 7.65 (s, 1H),7.57 (s, 1H), 7.39 (s, 1H), 7.14 (d, J = 8.2 Hz, 1H), 6.94 – 6.83 (m, 1H), 6.29 – 6.17 (m, 1H), 5.81 – 5.71 (m, 1H), 5.62 – 5.37 (m, 1H), 4.52 – 4.44(m, 1H), 4.14 – 3.77 (m, 3H), 3.56 – 3.47 (m, 1H), 3.36 – 3.33 (m, 1H), 3.28 – 3.22 (m, 2H), 1.14 – 0.96 (m, 4H).

[0179] Example 7: (R)-7'-(3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidium]-1'-one

[0180]

[0181] Step 1: Preparation of (R)-7'-(3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidium]-1'-one

[0182]

[0183] 7'-Chloro-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidine]-1'-one (83 mg, 0.397 mmol) was added to a microwave-safe tube, dissolved in 5 mL of 1,4-dioxane. Then, intermediate 2 (150 mg, 0.397 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (29 mg, 0.039 mmol), potassium carbonate (137 mg, 0.992 mmol), and water (1 mL) were added. The reaction was carried out under argon protection at 100 °C for 2 hours. After the reaction, the mixture was concentrated under reduced pressure, purified by column chromatography, and lyophilized to give 73 mg of the title product as a white solid. ESI-MS m / z: 423.9 [M+H] + . 1H NMR (400MHz, d6-DMSO) δ 8.48 (s, 2H), 8.21 (s, 1H), 8.02 (s, 2H), 7.46 (s, 1H), 6.93– 6.82 (m, 1H), 6.28 – 6.20 (m, 1H), 5.81 – 5.74 (m, 1H), 5.67 – 5.35 (m,1H), 4.53 – 4.44 (m, 1H), 4.03 – 3.73 (m, 3H), 3.57 – 3.47 (m, 1H), 3.35 –3.16 (m, 3H), 1.30 – 1.24 (m, 2H), 1.11 – 1.04 (m, 2H).

[0184] Example 8: (R)-7'-(3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)-2'-methyl-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidium]-1'-one

[0185]

[0186] Step 1: Preparation of 7'-chloro-2'-methyl-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidine]-1'-one

[0187]

[0188] 350 mg (1.677 mmol) of 7'-chloro-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidine]-1'-one was added to a two-necked flask and dissolved in N,N-dimethylformamide (6 mL). The mixture was purged three times with argon gas. Sodium hydride (100 mg, 2.516 mmol) was added at 0 °C, and the reaction was allowed to proceed for 10 minutes. Iodomethane (0.157 mL, 2.516 mmol) was then added, and the reaction was allowed to proceed for 2 hours at room temperature. After the reaction was complete, the mixture was quenched with saturated ammonium chloride, diluted with water, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to give 344 mg of the colorless oily title product. ESI-MS m / z: 223.0 [M+H] +

[0189] Step 2: Preparation of (R)-7'-(3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)-2'-methyl-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidium]-1'-one

[0190]

[0191] 7'-Chloro-2'-methyl-2',3'-dihydro-1'H-spiro[cyclopropane-1,4'-[2,6]naphthidine]-1'-one (118 mg, 0.529 mmol) was added to a microwave-safe tube, dissolved in 5 mL of 1,4-dioxane. Then, intermediate 2 (200 mg, 0.529 mmol), 1,1'-bis(diphenylphosphino)ferrocene palladium(II) dichloride (39 mg, 0.052 mmol), potassium carbonate (183 mg, 1.323 mmol), and water (1 mL) were added. The mixture was purged three times with argon gas, and the reaction was carried out at 100 °C for 2 hours using microwaves. After the reaction was complete, the mixture was concentrated under reduced pressure, purified by column chromatography, and lyophilized to give 50 mg of the title product as a white powder. ESI-MS m / z: 437.9 [M+H] + . 1 H NMR (400 MHz, d6-DMSO) δ 8.46 (s, 1H), 8.20 (s, 1H), 8.00 (s, 2H), 7.47(s, 1H), 6.93 – 6.83 (m, 1H), 6.25 (dd, J = 16.6, 1.9 Hz, 1H), 5.78 (dd, J =10.4, 1.7 Hz, 1H), 5.67 – 5.35 (m, 1H), 4.53 – 4.45 (m, 1H), 3.97 – 3.86 (m,1H), 3.85 – 3.74 (m, 1H), 3.58 – 3.49 (m, 1H), 3.47 (s, 2H), 3.38 (s, 2H), 3.06 (s, 3H), 1.32 – 1.24 (m, 2H), 1.14 – 1.08 (m, 2H).

[0192] Example 9: (R)-2'-(3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)-6',7'-dihydro-8'H-spiro[cyclopropane-1,5'-[1,7]naphthidium]-8'-one

[0193]

[0194] Step 1: Preparation of methyl 6-bromo-3-(bromomethyl)pyridinecarboxylate:

[0195]

[0196] Methyl 6-bromo-3-methylpyridinecarboxylate (10 g, 66.2 mmol) and N-bromosuccinimide (17.7 g, 99.3 mmol) were added to 100 mL of carbon tetrachloride solution, followed by the addition of azobisisobutyronitrile (1.085 g, 6.62 mmol). The mixture was purged with argon and reacted at 80 °C for 3 h. After the reaction was complete, the solution was concentrated, diluted with 100 mL of water, extracted with ethyl acetate, dried over anhydrous sodium sulfate, evaporated to dryness, and subjected to column chromatography to give the title product. ESI-MS m / z: 307.8, 309.9 [M+H] +

[0197] Step 2: Preparation of methyl 6-bromo-3-(cyanomethyl)pyridinecarboxylate

[0198]

[0199] Methyl 6-bromo-3-(bromomethyl)pyridinecarboxylate (1.75 g, 5.7 mmol) and trimethylcyanosilicon (0.78 ml, 6.23 mmol) were dissolved in acetonitrile (50 ml) and added to a single-necked flask. Tetrabutylammonium fluoride (8.6 ml, 8.55 mmol) was added dropwise at -20 °C, and the mixture was stirred at 0 °C for 3 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and subjected to column chromatography to give the title product. ESI-MS m / z: 254.9, 256.9 [M+H] +

[0200] Step 3: Preparation of methyl 6-bromo-3-(1-cyanocyclopropyl)pyridinecarboxylate

[0201]

[0202] Methyl 6-bromo-3-(cyanomethyl)pyridinecarboxylate (470 mg, 1.84 mmol) was dissolved in dimethyl sulfoxide (5 mL) in a single-necked flask. Potassium carbonate (382 mg, 2.03 mmol) and 1,2-dibromoethane (764 mg, 5.53 mmol) were added, and the mixture was stirred at 100 °C for 3 h. After the reaction was complete, the mixture was diluted with water, extracted with ethyl acetate, washed with saturated sodium chloride aqueous solution, dried over anhydrous sodium sulfate, and subjected to column chromatography to obtain the title product. ESI-MS m / z: 280.9, 282.9 [M+H] +

[0203] Step 4: Preparation of 2'-bromo-6',7'-dihydro-8'H-spiro[cyclopropane-1,5'-[1,7]naphthidine]-8'-one

[0204]

[0205] Methyl 6-bromo-3-(1-cyanocyclopropyl)pyridinecarboxylate (260 mg, 0.93 mmol) and cobalt chloride (480 mg, 3.7 mmol) were added to a three-necked flask, followed by the addition of methanol (5 mL). The mixture was purged with argon, and sodium borohydride (211 mg, 5.58 mmol) was added at 0 °C. The reaction proceeded to room temperature. After the reaction was complete, the mixture was concentrated under reduced pressure and subjected to column chromatography to obtain the title product. ESI-MS m / z: 252.9, 254.9 [M+H] +

[0206] Step 5: Preparation of (R)-2'-(3-(4-acrylomorpholin-3-yl)-5-chlorophenyl)-6',7'-dihydro-8'H-spiro[cyclopropane-1,5'-[1,7]naphthidium]-8'-one

[0207]

[0208] 2'-Bromo-6',7'-dihydro-8'H-spiro[cyclopropane-1,5'-[1,7]naphthidine]-8'-one (72 mg, 0.19 mmol), intermediate 2 (40 mg, 0.16 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (13 mg, 0.018 mmol), and potassium carbonate (45 mg, 0.38 mmol) were added to a microwave-safe tube, along with 5 mL of 1,4-dioxane and 1 mL of water. The reaction was carried out under argon protection at 100 °C for 2 h. After the reaction was complete, the mixture was concentrated under reduced pressure and separated to obtain 21 mg of a white solid. ESI-MS m / z: 423.9 [M+H] + . 1 H NMR (400 MHz, d6-DMSO) δ 8.35 (s, 1H), 8.10 (s,1H), 8.04 (d, J = 8.3 Hz, 1H), 8.00 (s, 1H), 7.61 (d, J = 8.3 Hz, 1H), 7.42(s, 1H), 6.94 – 6.79 (m, 1H), 6.29 – 6.18 (m, 1H), 5.81 – 5.73 (m, 1H), 5.66 – 5.41 (m, 1H), 4.53 – 4.45 (m, 1H), 4.29 – 4.02 (m, 1H), 3.93 – 3.75 (m,2H), 3.58 – 3.47 (m, 2H), 3.30 – 3.26 (m, 2H), 1.18 – 1.10 (m, 2H), 1.10 –1.01 (m, 2H).

[0209] Example 10: (S)-2-(3-((R)-4-acrylomorpholin-3-yl)-5-chlorophenyl)-6,6a,7,8-tetrahydro-9H-pyrido[3,4-b]pyrrolo[1,2-d][1,4]oxazin-9-one

[0210]

[0211] Step 1: Preparation of 4-bromo-6-chloropyridine-3-ol

[0212]

[0213] 4-Bromo-2-chloro-5-methoxypyridine (2.21 g, 10 mmol) was added to a reaction flask, purged three times with Ar gas, and then dichloromethane (20 mL) was added. Boron tribromide (2.89 mL, 30 mmol) was slowly added dropwise at 0 °C, and the reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS. After the reaction was complete, the reaction solution was quenched with methanol, and methanol was added dropwise until a solid precipitated. The mixture was filtered, the filter cake was washed with a small amount of methanol, and then dried to give 1.98 g of white solid. ESI-MS m / z: 207.8, 209.9 [M+H] + .

[0214] Step 2: Preparation of (S)-5-(((4-bromo-6-chloropyridin-3-yl)oxy)methyl)pyrrolidone-2-one

[0215]

[0216] 4-Bromo-6-chloropyridin-3-ol (2.07 g, 10 mmol) and (S)-5-(bromomethyl)pyrrolidone-2-one (1.95 g, 11 mmol) were dissolved in N,N-dimethylformamide (30 mL), followed by the addition of cesium carbonate (9.77 mg, 30 mmol). The reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS. After the reaction was complete, the solid was filtered off, and the reaction mixture was evaporated to dryness. The mixture was extracted three times with ethyl acetate, and the combined organic phases were washed with saturated sodium chloride and dried over anhydrous sodium sulfate. The mixture was then filtered, concentrated under reduced pressure, and yielded 2.696 g of crude yellow solid. ESI-MS m / z: 304.8, 306.8 [M+H] + .

[0217] Step 3: Preparation of (S)-2-chloro-6,6a,7,8-tetrahydro-9H-pyrido[3,4-b]pyrrolo[1,2-d][1,4]oxazin-9-one

[0218]

[0219] (S)-5-(((4-bromo-6-chloropyridin-3-yl)oxy)methyl)pyrrolidine-2-one (2.619 g, 8.62 mmol), tris(dibenzylindeneacetone)palladium (394.7 mg, 0.431 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (498.8 mg, 0.862 mmol), and cesium carbonate (8.4 g, 25.86 mmol) were added to a reaction flask, purged three times with Ar gas, and then 1,4-dioxane (60 mL) was added. The reaction system was stirred at 100 °C for 2 h. The reaction was monitored by LCMS. After the reaction was completed, the solid was filtered off, and the reaction solution was evaporated to dryness. The solution was extracted three times with EA, the organic phases were combined, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and column chromatography was performed to obtain 1.75 g of yellow-green solid.

[0220] ESI-MS m / z: 225.0 [M+H] + .

[0221] Step 4: Preparation of (S)-2-(3-((R)-4-acrylomorpholin-3-yl)-5-chlorophenyl)-6,6a,7,8-tetrahydro-9H-pyrido[3,4-b]pyrrolo[1,2-d][1,4]oxazin-9-one

[0222]

[0223] Intermediate 2 (170 mg, 0.45 mmol), (S)-2-chloro-6,6a,7,8-tetrahydro-9H-pyrido[3,4-b]pyrrolo[1,2-d][1,4]oxazin-9-one (100 mg, 0.45 mmol), [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (32.9 mg, 0.045 mmol), and potassium carbonate (124.2 mg, 0.9 mmol) were added to a mixed solvent of 1,4-dioxane and water (5 mL:1 mL). The mixture was reacted under argon protection at 100 °C for 2 h using microwave. After the reaction was complete as monitored by LC-MS, the product was purified by column chromatography, and 26 mg of the product was obtained by reverse-phase column purification. ESI-MS m / z: 439.9 [M+H] + . 1H NMR (400 MHz, d6-DMSO) δ8.78 (s, 1H), 8.35 (s, 1H), 7.82 (s, 1H), 7.78 (s, 1H), 7.45 (s, 1H), 6.93 –6.82 (m, 1H), 6.32 – 6.19 (m, 1H), 5.81 – 5.73 (m, 1H), 5.64 – 5.36 (m, 1H), 4.69 – 4.59 (m, 1H), 4.47 – 4.39 (m, 1H), 4.15 – 4.04 (m, 1H), 3.94 – 3.76(m, 4H), 3.58 – 3.45 (m, 1H), 3.26 – 2.86 (m, 1H), 2.76 – 2.59 (m, 1H), 2.46 – 2.34 (m, 1H), 2.30 – 2.17 (m, 1H), 1.83 – 1.65 (m, 1H).

[0224] Example 11: (R)-8-(3-(4-acryloylmorpholin-3-yl)-5-chlorophenyl)-3,4-dihydropyridino[4',3':4,5]pyrrolo[1,2-a]pyrazin-1(2H)-one

[0225]

[0226] Step 1: Preparation of ethyl 5-bromo-1-(2-(tert-butoxycarbonyl)amino)ethyl)-1H-pyrrolo[2,3-c]pyridine-2-carboxylate

[0227]

[0228] Ethyl 5-bromo-1H-pyrrolo[2,3-c]pyridine-2-carboxylate (2.0 g, 7.43 mmol, 1.0 eq), tert-butyl(2-bromoethyl)carbamate (2.17 g, 9.67 mmol, 1.3 eq), and potassium carbonate (3.08 g, 22.29 mmol, 3.0 eq) were dissolved in anhydrous N,N-dimethylformamide (15 mL), and the mixture was heated to 80 °C for 2 h. The reaction was monitored by LC-MS. After the reaction was complete, water was added, and the mixture was extracted three times with ethyl acetate. The organic phases were combined, washed three times with water, washed once with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to give 2.6 g of product. ESI-MS m / z: 412.1, 414.1 [M+H] + .

[0229] Step 2: Preparation of 5-bromo-1-(2-(tert-butoxycarbonyl)amino)ethyl)-1H-pyrrolo[2,3-c]pyridine-2-carboxylic acid

[0230]

[0231] Ethyl 5-bromo-1-(2-(tert-butoxycarbonyl)amino)ethyl)-1H-pyrrolo[2,3-c]pyridine-2-carboxylate (2.6 g, 6.33 mmol, 1.0 eq) was dissolved in ethanol (15 mL) and water (5 mL), and sodium hydroxide (760 mg, 18.99 mmol, 3.0 eq) was added. The reaction was carried out at 50 °C for 2 h. The reaction was monitored by LC-MS. After the reaction was completed, the pH was adjusted to weakly acidic with 1 N hydrochloric acid, and a white solid precipitated. The solid was filtered, the filter cake was collected, and the product was evaporated to dryness to give 2.2 g. ESI-MS m / z: 384.0, 386.0 [M+H] + .

[0232] Step 3: Preparation of 1-(2-aminoethyl)-5-bromo-1H-pyrrolo[2,3-c]pyridine-2-carboxylate

[0233]

[0234] 5-Bromo-1-(2-(tert-butoxycarbonyl)amino)ethyl)-1H-pyrrolo[2,3-c]pyridine-2-carboxylic acid was dissolved in 4 M dioxane hydrochloride solution and reacted at 50 °C for 2 h. The reaction was monitored by LC-MS. After the reaction was complete, the reaction solution was concentrated under reduced pressure to give 1.83 mg of yellow solid hydrochloride. ESI-MS m / z: 283.9, 285.9 [M+H] + .

[0235] Step 4: Preparation of 8-bromo-3,4-dihydropyridine[4',3':4,5]pyrrolo[1,2-a]pyrazin-1(2H)-one

[0236]

[0237] 1-(2-aminoethyl)-5-bromo-1H-pyrrolo[2,3-c]pyridine-2-carboxylate (2.0 g, 7.04 mmol, 1.0 eq) and N,N,N',N'-tetramethylchloroformamidine hexafluorophosphate (1.13 g, 14.08 mmol, 2.0 eq) were dissolved in acetonitrile (20 mL), and N-methylimidazolium (2.19 g, 26.75 mmol, 3.8 eq) was added. The reaction was carried out at room temperature for 2 h. The reaction was monitored by LC-MS. After the reaction was completed, the solution was concentrated and purified by column chromatography to give 2.8 g of white solid. ESI-MS m / z: 265.9, 267.9 [M+H] + .

[0238] Step 5: Preparation of (R)-8-(3-(4-acryloylmorpholin-3-yl)-5-chlorophenyl)-3,4-dihydropyridino[4',3':4,5]pyrrolo[1,2-a]pyrazin-1(2H)-one

[0239]

[0240] 8-Bromo-3,4-dihydropyrido[4',3':4,5]pyrrolo[1,2-a]pyrazin-1(2H)-one (120 mg, 0.4509 mmol) and intermediate 2 (170 mg, 0.4509 mmol) were added to a microwave-safe tube, dissolved in 1,4-dioxane (16 mL), followed by the addition of [1,1'-bis(diphenylphosphine)ferrocene]palladium dichloride (33 mg, 0.04509 mmol), potassium carbonate (187 mg, 1.353 mmol), and water (3.5 mL). The mixture was purged with argon, heated to 100 °C, and microwaved for 2 hours. After the reaction, the mixture was concentrated under reduced pressure and purified by column chromatography to obtain the title product (11 mg) as a white powder. ESI-MS m / z: 436.90 [M+H] + . 1H NMR (400 MHz, CD3OD_SPE) δ 8.89 (s, 1H), 8.07 – 7.90 (m, 3H), 7.31 (s,1H), 6.62 (s, 1H), 6.60 – 6.53 (m, 1H), 6.41 (d, J = 16.6 Hz, 1H), 5.78 (d, J= 10.4 Hz, 1H), 5.26 – 4.90 (m, 1H), 4.61 – 4.50 (m, 1H), 4.48 – 4.38 (m,2H), 4.03 – 3.95 (m, 1H), 3.95 – 3.83 (m, 3H), 3.68 – 3.59 (m, 1H), 3.66 –3.59 (m, 1H), 3.54 – 2.95 (m, 2H).

[0241] Example 12: (R)-3-(3-((R)-4-acryloylmorpholin-3-yl)-5-chlorophenyl)-6,6a,7,8-tetrahydro-9H-pyrido[4,3-b]pyrrolo[1,2-d][1,4]oxazin-9-one

[0242]

[0243] Step 1: Preparation of (R)-2-(((5-bromo-2-chloropyridin-4-yl)oxy)methyl)-5-oxopyrrolidine-1-carboxylic acid tert-butyl ester:

[0244]

[0245] 5-Bromo-2-chloro-4-hydroxypyridine (323 mg, 1.55 mmol), (2R)-2-hydroxymethyl-5-oxopyrrolidine-1-carboxylic acid tert-butyl ester (500 mg, 2.32 mmol), and triphenylphosphine (609 mg, 2.32 mmol) were placed in a two-necked flask, dissolved in tetrahydrofuran (25 mL), and the mixture was purged three times with argon gas and cooled to 0 °C. Diethyl azodicarbonate (404 mg, 2.32 mmol) was slowly added dropwise, and the mixture was stirred overnight at room temperature. After the reaction was complete, the mixture was concentrated and subjected to column chromatography to obtain the title product. ESI-MS m / z: 405.0, 407.0 [M+H] +

[0246] Step 2: Preparation of (R)-5-(((5-bromo-2-chloropyridin-4-yl)oxy)methyl)pyrrolidine-2-one

[0247]

[0248] (R)-2-(((5-bromo-2-chloropyridin-4-yl)oxy)methyl)-5-oxopyrrolidine-1-carboxylic acid tert-butyl ester (1 g, 2.47 mmol) was added to a single-necked flask, dissolved in dichloromethane (4 mL), followed by trifluoroacetic acid (2.5 mL). The mixture was stirred at room temperature for 2 h. After the reaction was complete, the pH was adjusted to weakly alkaline with saturated sodium bicarbonate solution, followed by extraction with dichloromethane, washing with saturated sodium chloride solution, drying over anhydrous sodium sulfate, and column chromatography to obtain the title product. ESI-MS m / z: 305.0, 307.0 [M+H] +

[0249] Step 3: Preparation of (R)-3-chloro-6,6a,7,8-tetrahydro-9H-pyrido[4,3-b]pyrrolo[1,2-d][1,4]oxazin-9-one

[0250]

[0251] (R)-5-(((5-bromo-2-chloropyridin-4-yl)oxy)methyl)pyrrolidine-2-one (1 g, 3.27 mmol) was dissolved in 1,4-dioxane (50 mL) in a two-necked flask. Tris(dibenzylacetone)dipalladium (302 mg, 0.33 mmol), 4,5-bis(diphenylphosphine-9,9-dimethyloxanthracene) (191 mg, 0.33 mmol), and cesium carbonate (2.13 g, 6.54 mmol) were added. After purging with argon three times, the mixture was stirred at 100 °C for 2 h. After the reaction was completed as monitored by LC-MS, the reaction solution was cooled to room temperature, quenched, and extracted three times with ethyl acetate. The organic layer was collected, washed with saturated sodium chloride, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and then subjected to column chromatography to obtain the title product. ESI-MS m / z: 225.0 [M+H] +

[0252] Step 4: Preparation of (R)-3-(3-((R)-4-acrylomorpholin-3-yl)-5-chlorophenyl)-6,6a,7,8-tetrahydro-9H-pyrido[4,3-b]pyrrolo[1,2-d][1,4]oxazin-9-one

[0253]

[0254] (R)-3-chloro-6,6a,7,8-tetrahydro-9H-pyrido[4,3-b]pyrrolo[1,2-d][1,4]oxazin-9-one (45 mg, 0.20 mmol), intermediate 2 (80 mg, 0.20 mmol), [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride (15 mg, 0.02 mmol), and potassium carbonate (55 mg, 0.42 mmol) were added to a microwave-safe tube. 5 mL of 1,4-dioxane and 1 mL of water were added. The mixture was microwave-safe at 100 °C for 2 h under argon protection. The reaction was monitored by LC-MS. After the reaction was complete, the mixture was concentrated under reduced pressure and separated to obtain 37 mg of the white solid title product. ESI-MS m / z: 440.0 [M+H] + . 1 H NMR (400 MHz, d6-DMSO) δ 9.50 (s, 1H), 8.06 – 7.94 (m, 2H), 7.64 (s, 1H), 7.37 (s, 1H), 6.96 –6.79 (m, 1H), 6.29 – 6.18 (m, 1H), 5.83 – 5.72 (m, 1H), 5.64 – 5.29 (m, 1H), 4.73 – 4.64 (m, 1H), 4.55 – 4.45 (m, 1H), 4.14 – 4.02 (m, 1H), 4.01 – 3.85(m, 3H), 3.83 – 3.74 (m, 1H), 3.57 – 3.46 (m, 1H), 3.32 – 3.16 (m, 1H), 2.74 – 2.59 (m, 1H), 2.44 – 2.35 (m, 1H), 2.32 – 2.22 (m, 1H), 1.86 – 1.65 (m,1H).

[0255] Example 13: (R)-1-(3-(3-chloro-5-(6-methyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']dipyridin-2-yl)phenyl)morpholinyl)prop-2-en-1-one

[0256]

[0257] Step 1: 2-Bromo-6-(2-(1-methylpiperidin-4-yl)hydrazino)pyridine

[0258]

[0259] 2-Bromo-6-hydrazinopyridine (5 g, 27 mmol) was dissolved in ethanol (15 mL), and 1-methylpiperidin-4-one (3 g, 27 mmol) was added. The reaction was carried out at 50 °C for 2 hours. After the reaction was completed, the solution was concentrated under reduced pressure to give 7 g of a brown powdery solid. ESI-MS m / z: 282.9, 284.9 [M+H] +

[0260] Step 2: Preparation of 2-bromo-6-methyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']bipyridine

[0261]

[0262] Weigh 5 g (23 mmol) of 2-bromo-6-(2-(1-methylpiperidin-4-yl)hydrazino)pyridine and add it to a solution at 180 °C. o In pre-stirred polyphosphoric acid at C, stir overnight at this temperature. After the reaction is complete, 90 o The precipitate was quenched slowly with 300 mL of water at C, and the pH was adjusted to 8 with sodium hydroxide. A large amount of brown solid precipitated out. The mixture was filtered, the filter cake was washed with plenty of water, collected, and dried under reduced pressure to obtain 3 g of powdery brown solid, which was directly added to the next step. ESI-MS m / z: 268.9, 270.9 [M+H] +

[0263] Step 3: Preparation of (R)-1-(3-(3-chloro-5-(6-methyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']dipyridin-2-yl)phenyl)morpholino)prop-2-en-1-one

[0264]

[0265] Intermediate 2 (170 mg, 0.451 mmol) was added to a microwave-safe tube, followed by 2-bromo-6-methyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']bipyridine (120 mg, 0.451 mmol), 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (33 mg, 0.05 mmol), and anhydrous potassium carbonate (125 mg, 0.902 mmol). The mixture was microwaved at 100 °C for 2 h. After the reaction was complete, the solution was evaporated to dryness and subjected to column chromatography to obtain 85 mg of a yellow solid. Further separation yielded 27 mg of the white solid title product. ESI-MS m / z: 436.9 [M+H] + . 1H NMR (400 MHz, d6-DMSO) δ 11.53 (s, 1H), 8.03 (s, 2H), 7.84 (d, J = 8.1 Hz, 1H), 7.61 (d, J = 8.1 Hz, 1H), 7.36 (s, 1H), 6.96 – 6.82 (m,1H), 6.30 – 6.18 (m, 1H), 5.82 – 5.72 (m, 1H), 5.67 – 5.33 (m, 1H), 4.49 (d,J = 12.3 Hz, 1H), 4.02 – 3.72 (m, 3H), 3.60 – 3.45 (m, 3H), 3.24 – 3.12 (m, 1H), 2.87 – 2.77 (m, 2H), 2.76 – 2.65 (m, 2H), 2.43 (s, 3H).

[0266] Example 14: (R)-1-(3-(3-chloro-5-(6,9-dimethyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']dipyridin-2-yl)phenyl)morpholinyl)prop-2-en-1-one

[0267]

[0268] Step 1: Preparation of 2-bromo-6,9-dimethyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']bipyridine

[0269]

[0270] 2-Bromo-6-methyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']bipyridine (200 mg, 0.752 mmol) was dissolved in DMF (3 ml), purged with argon, and sodium hydride (36 mg, 1.503 mmol) was added at 0 °C. After 15 min, dimethyl sulfate (95 mg, 0.752 mmol) was added, and the mixture was stirred at 0 °C for 2 h. After the reaction was complete, the mixture was quenched with saturated ammonium chloride and subjected to reverse-phase column chromatography to give 170 mg of a brown solid. ESI-MS m / z: 279.9, 282.0 [M+H] +

[0271] Step 2: Preparation of (R)-1-(3-(3-chloro-5-(6,9-dimethyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']dipyridin-2-yl)phenyl)morpholino)prop-2-en-1-one

[0272]

[0273] 2-Bromo-6,9-dimethyl-6,7,8,9-tetrahydro-5H-pyrrolo[2,3-b:4,5-c']bipyridine (95 mg, 0.339 mmol) was added to a microwave-safe tube, followed by the addition of 1,4-dioxane (5 ml) to dissolve it. Intermediate 2 (128 mg, 0.339 mmol), 1,1'-bis(diphenylphosphine)ferrocene palladium dichloride (25 mg, 0.034 mmol), and anhydrous potassium carbonate (94 mg, 0.068 mmol) were then added. The mixture was microwaved at 100 °C for 2 h. After the reaction was complete, the solution was evaporated to dryness and subjected to column chromatography to give 10 mg of the pale green solid title product. ESI-MS m / z: 450.9 [M+H] + . 1 H NMR (400 MHz, d6-DMSO) δ 8.11 (s, 1H), 8.07(s, 1H), 7.87 (d, J = 8.1 Hz, 1H), 7.65 (d, J = 8.2 Hz, 1H), 7.37 (s, 1H),6.96 – 6.88 (m, 1H), 6.30 – 6.21 (m, 1H), 5.82 – 5.75 (m, 1H), 5.64 – 5.46(m, 1H), 4.52 (m, J = 12.3 Hz, 1H), 3.95 – 3.78 (m, 3H), 3.73 (s, 3H), 3.58 –3.50 (m, 3H), 3.28 – 3.13 (m, 1H), 2.92 – 2.85 (m, 2H), 2.80 – 2.72 (m, 2H), 2.43 (s, 3H).

[0274] Following the synthesis methods described in the above embodiments, compounds from Examples 15-22 in Table 1 were prepared using different raw materials.

[0275] Table 1

[0276]

[0277] In vitro activity evaluation of compounds

[0278] Experimental Example 1: Compound-induced KEAP1-CUL3 protein interaction

[0279] The compounds of this invention promote the interaction between KEAP1 and CUL3 proteins, thereby increasing the ubiquitination and degradation of NRF2 protein. This invention verifies the ability of the compounds to induce the binding of KEAP1 and CUL3 proteins using a KEAP1-CUL3 HTRF assay.

[0280] 1. Experimental Materials

[0281] Compounds: The compounds of the present invention prepared in the above examples, and compound 96 disclosed in WO2024 / 073587. As a control compound (compound A), the preparation method of compound A is described in WO2024 / 073587, and it was identified by 1H NMR and mass spectrometry. Each compound was prepared into a 10 mM stock solution with DMSO and then successively diluted to 100,000 nM, 33,333.33 nM, 11,111.11 nM, 3,703.70 nM, 1,234.57 nM, 411.52 nM, 137.17 nM, 45.72 nM, 15.24 nM, 5.08 nM, 1.69 nM, and 0.56 nM.

[0282] Reagents: HTRF Mab Anti-FLAG M2-XL665, purchased from Revvity, Cat. No. 61FG2XLA; Streptavidin-Tb Cryptate, purchased from Revvity, Cat. No. 610SATLB; KEAP1-Biotin and CUL3-FLAG proteins, from Kanglong Chemical Co., Ltd.; HEPES pH 7.4, purchased from Invitrogen, Cat. No. 15630080; Pluronic F127, purchased from Sigma, Cat. No. P2443.

[0283] Experimental methods

[0284] 1.1 Buffer: 25 mM HEPES, 150 mM NaCl, 0.05% Pluronic F127, 100 μM Idiotheritiol (freshly prepared each time);

[0285] 1.2 The analyte was prepared with DMSO and serially diluted. The solution was added to the detection plate at 10 nL / well using Echo, and each concentration was repeated twice.

[0286] 1.3KEAP1-Biotin protein was diluted to 4 nM with buffer, and 1 μL / well was added to the detection plate;

[0287] 1.4 Centrifuge the test plate at 1000 rpm for 1 minute and incubate at 25 ºC for 3 hours;

[0288] 1.5 CUL3-FLAG protein, HTRF Mab Anti FLAG M2-XL665, and Streptavidin-TbCryptate reagent were mixed in equimolar amounts (150 nM), incubated at 25 ºC for 1 hour, and 3 μL / well was added to the detection plate.

[0289] 1.6 Centrifuge the detection plate at 1000 rpm for 1 minute, then incubate at 25 ºC for 1.5 hours.

[0290] 1.7 The detection plate was centrifuged at 1000 rpm for 1 minute, and the 665 nm / 616 nm ratio was detected by PHERAstar FSX.

[0291] 1.8 Data Analysis: Using DMSO wells as negative control wells and control compound A as positive control wells, the ratio of wells with the highest concentration of positive compound (100 μM) was 100%. The ratios of each compound in this invention were calculated. Delta F = 100% - (100% x (positive control well ratio - compound well ratio) / (positive control well ratio - negative control well ratio). The data were processed and fitted using XLfit to obtain IC50 values. 50 IC of the compound 50 The results are shown in Table 2.

[0292] Experimental Example 2: Compounds Inhibit the NRF2-ARE Signaling Pathway

[0293] Under normal physiological conditions, NRF2 binds to KEAP1 protein and is rapidly degraded via the CUL3-mediated ubiquitin-associated pathway. Under oxidative stress, KEAP1 Cys151 is covalently modified, dissociating KEAP1 from CUL3 and preventing NRF2 from being ubiquitinated. The homeostatically stabilized NRF2 protein translocates into the nucleus, forms a heterodimer with Maf, and activates ARE-dependent target gene expression. We constructed the KYSE70-ARE Luc Reporter cell and used a luciferase reporter gene assay to detect the ability of test compounds to promote Nrf2 protein degradation and inhibit its downstream ARE signaling pathway.

[0294] 1. Experimental Materials

[0295] Test compounds: The compounds of the present invention and control compound A prepared in the above examples were each prepared in 10 mM solution with DMSO and then diluted 5 times sequentially to 1000 nM, 200 nM, 40 nM, 8 nM, 1.6 nM, 0.32 nM, 0.064 nM, 0.0128 nM, and 0.00256 nM.

[0296] Cells: KYSE70 cells were purchased from Zhejiang Meisen Cell Technology Co., Ltd. (China).

[0297] Reagents: ARE-Luc Reporter Plasmid (purchased from Jiman Biotechnology, Cat. NO. GM-021048); RPMI 1640 (purchased from GIBCO, Cat. NO. 11875-093); FBS (purchased from GIBCO, Cat. NO. A5669701); Penicillin-Streptomycin (purchased from GIBCO, Cat. NO. 15140-122); Trypsin (purchased from GIBCO, Cat. NO. 25200072); GMOne-Step Luciferase Reporter GeneAssay Kit (purchased from Jiman Biotechnology, Cat. NO. GM-040503C); Star 96-well plates (purchased from Corning, Cat. NO. 3599).

[0298] 2. Experimental Methods

[0299] 2.1 KYSE70 cells were transfected with ARE-Luc Reporter Plasmid, and single clones were selected to obtain KYSE70-ARELuc Reporter cells;

[0300] 2.2 Collect and count KYSE70-ARE Luc Reporter cells in the logarithmic growth phase at a concentration of 1.5 × 10⁻⁶. 4 Cells were seeded in 96-well plates and incubated overnight at 37°C in a cell culture incubator.

[0301] 2.3 The test compound was prepared into a 10 mM stock solution using DMSO, and then successively diluted 5-fold with RPMI 1640 medium containing 1% FBS to prepare drug-containing medium solutions of 1000 nM, 200 nM, 40 nM, 8 nM, 1.6 nM, 0.32 nM, 0.064 nM, 0.0128 nM, and 0.00256 nM.

[0302] 2.4 Discard the original culture medium in the 96-well plate, add the corresponding drug-containing culture medium at 100 μL / well, and incubate in a 37℃ cell incubator for 24 h;

[0303] Add GMOne-Step Luciferase Reporter Gene Assay Kit to 2.596 wells, mix thoroughly, and incubate at room temperature for 5 minutes;

[0304] 2.6 Take 100 μL of supernatant and detect the bioluminescence signal in each well (reading condition: 500 ms).

[0305] 2.7 Using Prism GraghPad software, curves were plotted and fitted to obtain the IC50 values ​​for each compound. 50 The experimental results are shown in Table 2.

[0306] Table 2

[0307]

[0308] The experiments above demonstrate that the compound of this invention can activate the KEAP1 protein and increase its interaction with the CUL3 protein, thereby inducing the degradation of the NRF2 protein, making it a promising small molecule anti-tumor drug.

[0309] Although the present invention has been described in detail above, those skilled in the art will understand that various modifications and changes can be made to the invention without departing from its spirit and scope. The scope of the invention is not limited to the detailed description above, but should be attributed to the claims.

Claims

1. A compound of general formula (I) or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, or a prodrug, in, X is selected from O, CH2, and NH; L is selected from single bond, -C 1-6 Alkyl groups, -NH groups, -O groups, and -S groups; Ring A is selected from aryl, heteroaryl, cycloalkyl, and heterocyclic groups; R 1 R 2 R 3 Each is independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino; R 4 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkyl aminoacyl group, oxo group, double C 1-6 Alkylamino, aryl, heteroaryl, cycloalkyl, and heterocyclic groups; and m, n, and q are each independently selected from 0, 1, 2, 3, and 4.

2. The compound or isomer thereof, pharmaceutically acceptable salt, solvate, crystal or prodrug according to claim 1, wherein ring A is selected from C. 6-15 Aryl, 5-15 quinone heteroaryl, C 3-15 Cycloalkyl groups and 3-15 membered heterocyclic groups, wherein the heteroatoms are selected from N, O, S, P and B.

3. The compound or isomer thereof, pharmaceutically acceptable salt, solvate, crystal or prodrug according to claim 1 or 2, wherein R 4 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkyl aminoacyl group, oxo group, double C 1-6 Alkylamino, C 6-15 Aryl, 5-15 quinone heteroaryl, C 3-15 Cycloalkyl and 3-15 membered heterocyclic groups.

4. The compound or isomer thereof, pharmaceutically acceptable salt, solvate, crystal or prodrug according to claim 1 or 2, wherein general formula (I) has the structure of general formula (Ib): in, Ring B and ring C are each independently selected from C. 6-12 Aryl, 5-8 quinone heteroaryl, C 3-8 Cycloalkyl and 3-8 membered heterocyclic groups, optionally surrounded by one or more groups selected from halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 The alkylamino group is substituted, wherein the heteroatom is selected from N, O, S and P; Rings B and C can be either parallel rings or spiral rings; Y1 and Y2 are each independently selected from CH and N; X, L, R 1 R 2 R 3 , m and n have the definitions described in general formula (I).

5. The compound or isomer thereof, pharmaceutically acceptable salt, solvate, crystal or prodrug according to claim 4, wherein the group Selected from , , , , , , , , and .

6. A compound of general formula (II) or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug, in, Ring B and ring C are each independently selected from C. 6-15 Aryl, 5-15 quinone heteroaryl, C 3-15 Cycloalkyl and 3-15 membered heterocyclic groups, optionally surrounded by one or more groups selected from halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino group substitution; R 5 R 6 R 7 Each is independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino; Y is selected from O, CH2, and NH; and r and s are each independently selected from 0, 1, 2, 3 and 4.

7. The compound or isomer thereof, pharmaceutically acceptable salt, solvate, crystal or prodrug according to claim 6, wherein ring B is selected from pyrimidine, optionally surrounded by one or more elements selected from halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino group substitution; and Ring C is selected from pyrazole, pyridine, , and It is optionally mixed with one or more elements selected from halogen, hydroxyl, C 1-6 Alkyl, C 2-4 alkenyl, C 2-4 alkynyl, 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, C 1-6 alkylsulfonyl, aminoacyl, C 1-6 Alkylaminoacyl, oxo group and double C 1-6 Alkylamino group substitution.

8. A compound or an isomer thereof, a pharmaceutically acceptable salt, a solvate, a crystal, or a prodrug, wherein said compound is selected from the group consisting of: ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， ， , , and 。 9. A pharmaceutical composition comprising a compound or an isomer thereof as claimed in any one of claims 1-8, a pharmaceutically acceptable salt, a solvate, a crystal or prodrug, and a pharmaceutically acceptable carrier.

10. The use of the compound or isomer thereof, pharmaceutically acceptable salt, solvate, crystal or prodrug, or pharmaceutical composition of any one of claims 1-8 in the preparation of a medicament for treating KEAP1-mediated diseases.

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