GSPT1 degradation agent and application thereof

Abstract

The invention belongs to the field of medical chemistry, and particularly relates to compounds serving as GSPT1 degradation agents or isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs of the compounds, preparation methods of the compounds, pharmaceutical compositions containing the compounds and application of the compounds or the compositions to treatment of diseases beneficial to GSPT1 degradation, such as cancers. # imgabs0 #

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry, and specifically relates to compounds or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs as GSPT1 degraders, their preparation methods, and pharmaceutical compositions containing these compounds and the use of these compounds or compositions for treating diseases such as cancer that are beneficial for degrading GSPT1. Background Art

[0002] Cereblon (CRBN) is a protein encoded by the CRBN gene. Cereblon forms an E3 ubiquitin ligase complex with damaged DNA binding protein 1 (DDB1), Cullin-4A (CUL4A), and regulator of cullins 1 (ROC1). This complex can ubiquitinate many other proteins.

[0003] Cereblon, as a component of the DDB1-CUL4a-Roc1 ubiquitin ligase complex, is a molecular target of immunomodulators such as thalidomide, lenalidomide, and pomalidomide. Inhibition of CRBN ubiquitination by these drugs can lead to CRBN accumulation, resulting in increased degradation of target proteins mediated by cullin-4RING E3 ligase. Cereblon E3 ligase inhibitors have a clear mechanism in the treatment of tumor-related diseases.

[0004] GSPT1 (G1 to S phase transition 1, also known as eRF3a) plays an important role in inhibiting cell proliferation by interacting with the release factor eRF1, mediating stop codon recognition and the release of nascent proteins from ribosomes. Studies have shown that GSPT1, as a new substrate of CRBN, can be effectively degraded by CRBN protein regulators, resulting in significant inhibition of leukemia cell proliferation. It has also been reported that GSPT1 is a cause of off-target effects of PTOTAC molecules that recruit CRBN. As a target of molecular glue, GSPT1 plays an important role in the protein translation process. Its degradation can lead to abnormal protein synthesis, thereby affecting the proliferation and survival of tumor cells. At the same time, as a GTPase, GSPT1 is difficult to target because the intracellular GTP concentration is very high and the GTP binding pocket binds strongly to the GTPase.

[0005] Safe and effective GSPT1 degraders need to be developed to meet clinical needs. Summary of the invention

[0006] One object of the present invention is to provide a class of compounds having GSPT1 degradation activity represented by general formula (I) or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs,

[0007]

[0008] Another object of the present invention is to provide a method for preparing the compound of general formula (I) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug.

[0009] Another object of the present invention is to provide a composition comprising a compound of the general formula (I) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and a pharmaceutically acceptable carrier, as well as a composition comprising a compound of the general formula (I) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug and another one or more drugs.

[0010] Another object of the present invention is to provide a method for treating GSPT1-related diseases such as cancer using the compound of the general formula (I) of the present invention or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs, and use of the compound of the general formula (I) of the present invention or its isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs in the preparation of drugs for treating GSPT1-related diseases such as cancer.

[0011] In view of the above-mentioned invention object, the present invention provides the following technical solutions:

[0012] In a first aspect, the present invention provides a compound represented by general formula (I) or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof,

[0013]

[0014] in,

[0015] X 1 is selected from -CH- and -N-;

[0016] R 1 , R 2 , R 3 , R 4 are each independently selected from hydrogen, halogen, alkyl and haloalkyl;

[0017] R 5 is selected from hydrogen and alkyl;

[0018] R 6is selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, cycloalkyl and heterocyclyl; which is optionally substituted with halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl;

[0019] L 1 , L 2 are each independently selected from a bond, -CH 2 -, -O-, -NH-, and -C(O)-;

[0020] L 3 Selected from bonds and -C≡C-;

[0021] Cy is selected from aryl and heteroaryl;

[0022] Y is selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, optionally substituted with halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, and alkylaminoacyl;

[0023] n is selected from 0, 1, 2, 3, 4 and 5; and

[0024] When X 1 When selected from -CH-, L 3 Selected from -C≡C-.

[0025] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0026] R 1 , R 2 , R 3 , R 4 are each independently selected from hydrogen, halogen, C 1-6 Alkyl and halogenated C 1-6 alkyl;

[0027] More preferably, R 1 , R 2 , R 3 , R4 are each independently selected from hydrogen, halogen, C 1-3 Alkyl and halogenated C 1-3 alkyl;

[0028] More preferably, R 1 , R 2 , R 3 , R 4 Each is independently selected from hydrogen, fluorine, chlorine, bromine, iodine, methyl, ethyl, propyl, isopropyl, isobutyl, trifluoromethyl and trifluoroethyl.

[0029] In some specific embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein: R 1 , R 4 Selected from fluorine; R 2 , R 3 Selected from hydrogen.

[0030] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0031] R 5 Selected from hydrogen and C 1-6 alkyl;

[0032] More preferably, R 5 Selected from hydrogen and C 1-3 alkyl;

[0033] More preferably, R 5 Selected from hydrogen, methyl, ethyl, propyl and isopropyl.

[0034] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0035] R 6 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halo C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C1-6 Alkylamino, C 3-8 Cycloalkyl and 3-8 membered heterocyclic groups, which are optionally substituted by halogen, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, C 3-8 Cycloalkyl, C 3-8 Heterocycloalkyl, C 6-12 Aryl and C 5-12 heteroaryl substitution;

[0036] More preferably, R 6 Selected from hydrogen, halogen, hydroxyl, C 1-3 Alkyl, C 2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino, C 3-6 Cycloalkyl and 3-6 membered heterocyclic groups, which are optionally substituted by halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, C 3-8 Cycloalkyl, C 3-8 Heterocycloalkyl and C 6-12 Aryl substitution;

[0037] More preferably, R 6is selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxy, methyl, ethyl, propyl, isopropyl, vinyl, ethynyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino, C 3-6 Cycloalkyl and 3-6 membered heterocyclic groups, which are optionally substituted by halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, C 3-8 Cycloalkyl, C 3-8 Heterocycloalkyl, C 6-12 Aryl and C 5-12 Heteroaryl substitution.

[0038] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0039] L 1 , L 2 Each independently selected from -CH 2 -, -O-, -NH-, and -C(O)-;

[0040] More preferably, L 1 Selected from -O- and -NH-; L 2 is selected from -O-, -NH- and -C(O)-.

[0041] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0042] Cy is selected from C 6-10 Aryl and 5-12 membered heteroaryl;

[0043] Further preferably, Cy is selected from C 6-8 Aryl and 5-10 membered heteroaryl;

[0044] Still further preferably, Cy is selected from phenyl, imidazolyl, pyrrolyl, furanyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, indolyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, benzopyrazolyl, benzimidazolyl, benzofuranyl, benzopyranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, quinolyl, isoquinolyl, quinazolinyl, cinnolinyl, quinoxalinyl, benzoxazinyl, benzothiazinyl, imidazopyridinyl, pyrimidopyrazolyl, pyrimidoimidazolyl.

[0045] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0046] Y is selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halo C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 3-8 Heterocycloalkyl, C 6-12 Aryl and C 5-12 Heteroaryl, which is optionally substituted by halogen, hydroxy, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl substitution;

[0047] More preferably, Y is selected from hydrogen, halogen, hydroxyl, C 1-3 Alkyl, C2-4 Alkenyl, C 2-4 Alkynyl, halo C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino, C 3-8 Cycloalkyl, C 3-8 Oxyheterocycloalkyl, C 3-8 Azacycloalkyl, C 6-12 Aryl and C 5-12 Heteroaryl, which is optionally substituted by halogen, hydroxy, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl and C 1-3 Alkylaminoacyl substitution;

[0048] More preferably, Y is selected from hydrogen, halogen, hydroxy, methyl, ethyl, propyl, isopropyl, fluorine, chlorine, bromine, iodine, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, halogenated C 1-3 Alkyl, vinyl, ethynyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl, C 1-3 Alkylaminoacyl, di-C 1-3 Alkylamino, cyclopropyl, cyclobutyl, cyclopentyl, oxacyclopropyl, oxetanyl, oxacyclopentyl, phenyl, pyrazolyl, pyridyl, C 3-8 Azoheterocycloalkyl such as 7-oxa-2-azaspiro[3.5]non-2-yl, C 6-12 Aryl and C 5-12Heteroaryl, which is optionally substituted with fluorine, chlorine, hydroxy, methyl, ethyl, propyl, isopropyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl and C 1-3 Alkylaminoacyl substitution.

[0049] In some embodiments, the compound of the present invention is a compound of formula (I) or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof, wherein:

[0050] n is selected from 0, 1, 2, 3 and 4;

[0051] Further preferably, n is selected from 0, 1, 2 and 3;

[0052] More preferably, n is selected from 0, 1 and 2.

[0053] In some embodiments, the present invention provides a compound represented by the general formula (I) or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof, wherein the general formula (I) has the structure of the following general formula (Ia),

[0054]

[0055] Among them, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , L 1 , L 2 , L 3 , Y and n have the same meanings as described above for the general formula (I).

[0056] In some embodiments, the compound of the general formula (Ia) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein R 6 is selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxy, methyl, ethyl, propyl, isopropyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, n is selected from 0, 1 and 2.

[0057] In some embodiments, the compound of the general formula (Ia) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Y is selected from hydrogen, methyl, ethyl, propyl, isopropyl, It is optionally replaced by one or more hydrogen, halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl and C 1-3 Alkylaminoacyl substitution.

[0058] In some specific embodiments, the compound of the general formula (Ia) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Selected from

[0059] In some embodiments, the present invention provides a compound represented by formula (Ib) or a pharmaceutically acceptable salt, isomer, solvate, crystal or prodrug thereof,

[0060]

[0061] Among them, X 1 , R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , L 1 , L 2 , Y and n have the same meanings as described above for the general formula (I).

[0062] In some embodiments, the compound of the general formula (Ib) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein R 6 is selected from hydrogen, fluorine, chlorine, bromine, iodine, hydroxy, methyl, ethyl, propyl, isopropyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclyl; n is selected from 0, 1, 2 and 3.

[0063] In some embodiments, the compound of the general formula (Ib) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Y is selected from hydrogen, methyl, ethyl, propyl, isopropyl, It is optionally replaced by one or more hydrogen, halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl and C 1-3 Alkylaminoacyl substitution.

[0064] In some specific embodiments, the compound of the general formula (Ib) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Selected from

[0065] The present invention provides the following specific compounds or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs:

[0066]

[0067]

[0068] In a second aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof.

[0069] In some embodiments, the present invention provides a compound of the present invention or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof, and a pharmaceutical composition comprising the compound of the present invention or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal or a prodrug thereof, wherein the compound or the pharmaceutical composition is used to treat a GSPT1-related disease such as cancer.

[0070] In some embodiments, the present invention provides a pharmaceutical composition comprising a compound of the present invention or an isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug thereof and a pharmaceutically acceptable carrier.

[0071] The compound of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug can be mixed with a pharmaceutically acceptable carrier, diluent or excipient to prepare a pharmaceutical preparation, suitable for oral or parenteral administration. The method of administration includes, but is not limited to intradermal, intramuscular, intraperitoneal, intravenous, subcutaneous, intranasal and oral routes. The preparation can be administered by any route, such as by infusion or push injection, by the route of absorption through the epithelium or skin mucosa (such as oral mucosa or rectum, etc.). Administration can be systemic or local. Examples of oral administration preparations include solid or liquid dosage forms, specifically, tablets, pills, granules, powders, capsules, syrups, emulsions, suspensions, etc. The preparation can be prepared by methods known in the art, and includes carriers, diluents or excipients conventionally used in the field of pharmaceutical preparations.

[0072] In a third aspect, the present invention provides a method for treating GSPT1-related diseases using a compound represented by formula (I), (Ia), (Ib) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, or a pharmaceutical composition comprising the same, as well as use of the compound in the preparation of a drug for treating GSPT1-related diseases.

[0073] In some preferred embodiments, the present invention provides a method for treating GSPT1-related diseases such as cancer using a compound represented by formula (I), (Ia), (Ib) of the present invention or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, or a pharmaceutical composition comprising the same, as well as use in the preparation of a drug for treating GSPT1-related diseases such as cancer.

[0074] In some preferred embodiments, the present invention provides a compound represented by the general formula (I), (Ia), or (Ib) of the present invention, or an isomer, a pharmaceutically acceptable salt, a solvate, a crystal, or a prodrug thereof, or a pharmaceutical composition comprising the same for use in a method for treating cancer and in the preparation of a drug for treating cancer, wherein the cancer includes, but is not limited to, non-small cell lung cancer, lung adenocarcinoma, lung cancer, large cell lung cancer, non-small cell lung cancer, lung squamous cell carcinoma, small cell lung cancer, melanoma, desmoplastic melanoma, uveal melanoma, ovarian small cell carcinoma, ovarian rhabdoid tumor, skin squamous cell carcinoma, glioma, uterine cancer, endometrial cancer, ovarian serous cystadenocarcinoma, bladder urothelial carcinoma, primary central nervous system lymphoma, esophageal cancer, bladder cancer, gastric adenocarcinoma, adenoid cystic carcinoma, lymphoid tumor diffuse large B-cell lymphoma, pancreatic cancer, colorectal adenocarcinoma, gallbladder cancer, Tube cancer, sarcoma, head and neck cancer, cervical cancer and endocervical cancer, medulloblastoma, cutaneous T-cell lymphoma, hepatocellular carcinoma, renal papillary cell carcinoma, breast cancer, mantle cell lymphoma, gallbladder cancer, testicular germ cell cancer, renal clear cell carcinoma, prostate cancer, pediatric Ewing's sarcoma, thymoma, renal chromophobe cell carcinoma, renal non-clear cell carcinoma, pheochromocytoma and paraganglioma, thyroid cancer, malignant peripheral nerve sheath tumor, neuroendocrine prostate cancer, head Squamous cell carcinoma of the neck, adrenal cortical carcinoma, cervical cancer and endocervical cancer, squamous cell carcinoma of the skin, testicular germ cell carcinoma, glioblastoma, glioblastoma multiforme, Ewing sarcoma, clear cell renal cell carcinoma, neuroblastoma, diffuse large B-cell lymphoma, acute myeloid leukemia, chronic lymphocytic leukemia, multiple myeloma, malignant rhabdoid tumor, epithelioid sarcoma, renal medullary carcinoma, synovial sarcoma, meningioma, and soft tissue sarcoma.

[0075] In some specific embodiments, the compounds represented by the general formula (I), (Ia), (Ib) of the present invention or their isomers, pharmaceutically acceptable salts, solvates, crystals or prodrugs are used to treat diseases caused by accumulation of GSPT1 protein, such as non-Hodgkin's lymphoma, leukemia and / or solid tumors. The non-Hodgkin's lymphoma is selected from one or more of diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), follicular lymphoma, mucosa-associated lymphoid tissue lymphoma and T-cell lymphoma. The leukemia includes one or more of chronic lymphocytic leukemia, chronic myeloid leukemia, acute lymphocytic leukemia and acute myeloid leukemia. The solid tumor includes one or more of lung cancer, liver cancer, breast cancer and brain glioma.

[0076] Definition of terms

[0077] Unless stated otherwise, the terms used in the specification and claims have the following meanings.

[0078] The "hydrogen", "carbon" and "oxygen" in the compounds of the present invention include all isotopes thereof. Isotopes should be understood to include those atoms having the same atomic number but different mass numbers. For example, isotopes of hydrogen include protium, tritium and deuterium, and isotopes of carbon include 12 C. 13 C and 14 C, oxygen isotopes include 16 O and 18 O etc.

[0079] The "isomer" of the present invention refers to a molecule with the same atomic composition and connection mode, but different three-dimensional spatial arrangement, 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, that is, they have the ability to rotate the plane of plane polarized light. When describing optically active compounds, the prefix D, L or R, S is used to indicate the absolute configuration of the chiral center of the molecule. The prefix D, L or (+), (-) is used to name the sign of rotation of plane polarized light of the compound, (-) or L means that the compound is left-handed, and the prefix (+) or D means that the compound is right-handed. The chemical structure of these stereoisomers is the same, but their stereostructures are different. Specific stereoisomers can be enantiomers, and a mixture of isomers is usually called an enantiomeric mixture. A 50:50 mixture of enantiomers is called a racemic mixture or a racemate, which may result in no stereoselectivity or stereospecificity during chemical reactions. The terms "racemic mixture" and "racemate" refer to an equimolar mixture of two enantiomers, devoid of optical activity.

[0080] Depending on the choice of starting materials and process, the compounds of the invention may exist in the form of one of the possible isomers or a mixture thereof, such as a racemate and a diastereomeric mixture (depending on the number of asymmetric carbon atoms). Optically active (R)- or (S)-isomers may be prepared using chiral synthons or chiral reagents, or resolved using conventional techniques.

[0081] Any resulting mixture of stereoisomers can be separated into the pure or substantially pure geometric isomers, enantiomers, diastereomers on the basis of the differences in the constituent physicochemical properties, for example, by chromatography and / or fractional crystallization.

[0082] The "halogen" of the present invention refers to fluorine, chlorine, bromine, and iodine. The "halogenated" of the present invention refers to substitution with fluorine, chlorine, bromine, or iodine.

[0083] The "alkyl" of the present invention refers to a straight or branched saturated aliphatic hydrocarbon group, preferably a straight or branched group containing 1 to 6 carbon atoms, and further preferably a straight or branched group containing 1 to 3 carbon atoms, non-limiting examples of which include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be at any available point of attachment.

[0084] The "alkenyl" of the present invention refers to a straight or branched hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms, containing at least one carbon-carbon double bond, and having 2 to 12 carbon atoms. In some embodiments, the alkenyl group contains 2 to 8 carbon atoms. In other embodiments, the alkenyl group contains 2 to 4 carbon atoms. The alkenyl group is connected to the rest of the molecule by a single bond, such as vinyl, allyl, but-1-enyl, pent-1-enyl, pent-1,4-dienyl, etc. The alkenyl group is optionally substituted with one or more other substituents, such as alkyl, halogen, cyano, nitro, oxygen, sulfur, imino, haloalkyl, alkoxy, cycloalkyl, heterocyclic radical, heteroaryl, heteroaryl, etc.

[0085] The "alkynyl" of the present invention refers to a straight or branched hydrocarbon chain radical consisting only of carbon atoms and hydrogen atoms, containing at least one carbon-carbon triple bond, and having 2 to 12 carbon atoms. In some embodiments, the alkynyl group comprises 2 to 8 carbon atoms. In other embodiments, the alkynyl group comprises 2 to 4 carbon atoms. The alkynyl group is connected to the rest of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc. The alkynyl group is optionally substituted with one or more other substituents, such as alkyl, halogen, cyano, nitro, oxygen, sulfur, imino, haloalkyl, alkoxy, cycloalkyl, heterocyclic, heteroaryl, heteroaryl, etc.

[0086] The -C(O)- group of the present invention means a "carbonyl group".

[0087] The "haloalkyl group" of the present invention refers to an alkyl group substituted with at least one halogen.

[0088] The "hydroxyalkyl group" of the present invention refers to an alkyl group substituted with at least one hydroxy group.

[0089] The "alkoxy" of the present invention refers to -O-alkyl. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, n-propoxy, isopropoxy, isobutoxy, sec-butoxy, etc. Alkoxy can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any available point of attachment.

[0090] The "cycloalkyl" of the present invention refers to a cyclic saturated hydrocarbon group. Suitable cycloalkyl groups may be substituted or unsubstituted monocyclic, bicyclic or tricyclic saturated hydrocarbon groups having 3 to 12 carbon atoms, such as cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl.

[0091] The "heterocyclyl" of the present invention refers to a group of a 3- to 12-membered non-aromatic ring system having 1 to 4 ring heteroatoms, each of which is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus and silicon ("3-12 membered heterocyclyl"). In heterocyclyl groups containing one or more nitrogen atoms, the point of attachment can be a carbon or nitrogen atom, as long as the valence permits. The heterocyclyl group can be either monocyclic ("monocyclic heterocyclyl") or a fused, bridged or spiral ring system (e.g., a bicyclic system (also known as a "bicyclic heterocyclyl")) and can be saturated or partially unsaturated. Suitable heterocyclyls include, but are not limited to, piperidinyl, azetidinyl, aziridine, tetrahydropyrrolyl, piperazinyl, dihydroquinazolinyl, oxacyclopropyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, etc. Each example of a heterocyclyl group can be optionally substituted or unsubstituted, and when substituted, the substituent can be at any usable point of attachment.

[0092] The "aryl" of the present invention refers to an aromatic system that may contain a monocyclic or fused polycyclic ring, preferably a monocyclic or fused bicyclic ring, containing 6 to 12 carbon atoms, preferably containing about 6 to about 10 carbon atoms. Suitable aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, fluorenyl, indanyl. Aryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any usable point of attachment.

[0093] The "heteroaryl" or "heteroaromatic ring" of the present invention refers to an aromatic group in which at least one carbon atom is replaced by a heteroatom, preferably consisting of 5-12 atoms (5-12-membered heteroaryl), and more preferably consisting of 5-10 atoms (5-10-membered heteroaryl), wherein the heteroatom is O, S, or N. The heteroaryl groups include, but are not limited to, imidazolyl, pyrrolyl, furanyl, thienyl, pyrazolyl, oxazolyl, thiazolyl, isoxazolyl, isothiazolyl, oxadiazolyl, triazolyl, tetrazolyl, indolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, triazinyl, isoindolyl, benzopyrazolyl, benzimidazolyl, benzofuranyl, benzopyranyl, benzothienyl, benzoxazolyl, benzothiazolyl, benzisoxazolyl, benzisothiazolyl, quinolyl, isoquinolyl, quinazolinyl, cinnolinyl, quinoxalinyl, benzoxazinyl, benzothiazinyl, imidazopyridinyl, pyrimidopyrazolyl, pyrimidoimidazolyl, etc. The heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents may be at any available point of attachment.

[0094] The "pharmaceutically acceptable salt" of the present invention refers to the salt of the compound of the present invention, which is safe and effective when used in mammals and has the desired biological activity.

[0095] The "solvate" of the present invention refers to a complex formed by the combination of a solute (such as an active compound, a salt of an active compound) and a solvent (such as water) in a conventional sense. The solvent refers to a solvent known to or easily determined by a person skilled in the art. If it is water, the solvate is usually referred to as a hydrate, such as a hemihydrate, a monohydrate, a dihydrate, a trihydrate or an alternative thereof.

[0096] The in vivo effects of the compounds of formula (I) may be partially exerted by one or more metabolites formed in the human or animal body after the compound of formula (I) is administered. As described above, the in vivo effects of the compounds of formula (I) may also be exerted via the metabolism of precursor compounds ("prodrugs"). "Prodrugs" of the present invention refer to compounds that are converted into compounds of the present invention under physiological conditions in an organism due to reactions with enzymes, gastric acid, etc., i.e., compounds that are converted into compounds of the present invention by oxidation, reduction, hydrolysis, etc. of enzymes and / or compounds that are converted into compounds of the present invention by hydrolysis reactions of gastric acid, etc.

[0097] The term "crystalline" in the present invention refers to a solid whose internal structure is formed by regularly repeating constituent atoms (or groups thereof) in three dimensions, and is distinguished from an amorphous solid that does not have such a regular internal structure.

[0098] The "pharmaceutical composition" of the present invention refers to a mixture comprising any one of the compounds of the present invention, including corresponding isomers, prodrugs, solvates, pharmaceutically acceptable salts or chemically protected forms thereof, and one or more pharmaceutically acceptable carriers and / or one or more other drugs. The purpose of the pharmaceutical composition is to facilitate the administration of the compound to an organism. The composition is generally used to prepare a drug for the treatment and / or prevention of a disease mediated by one or more kinases.

[0099] The "pharmaceutically acceptable carrier" of the present invention refers to a carrier that does not cause significant irritation to an organism and does not interfere with the biological activity and properties of the administered compound, including 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 compound of the present 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 carboxymethylcellulose, as well as cellulose and cellulose acetate; malt, gelatin, etc.

[0100] The "excipient" of the present invention refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Excipients may include calcium carbonate, calcium phosphate, various sugars and various types of starch, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols. DETAILED DESCRIPTION

[0101] The present invention is further described in detail below in conjunction with the examples, but the present invention is not limited to these examples. The materials used in the following examples are commercially available unless otherwise specified.

[0102] Example 1: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate

[0103]

[0104] Step 1: Synthesis of ethyl 3-((4-bromo-2,6-difluorophenyl)amino)propanoate

[0105]

[0106] 4-Bromo-2,6-difluoroaniline (20.0 g, 96.1 mmol) was added to a single-necked flask, ethyl acrylate (100 mL) was added to dissolve it, and then trifluoromethanesulfonic acid (720 mg, 4.8 mmol) was added and reacted at 95°C for 12 h. After the reaction was completed, the reaction solution was concentrated and column chromatography was performed to obtain the title product. ESI-MS m / z: 307.9 [M+H] + .

[0107] Step 2: Synthesis of 1-(4-bromo-2,6-difluorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0108]

[0109] Add methyl 5-bromo-2-(bromomethyl)benzoate (10.3g, 33.4mmol) to tetrahydrofuran (15mL), protect with inert gas, ice bath to 0℃, then add 2,2,2-trichloroacetyl isocyanate (6.91g, 36.7mmol), react at 0℃ for 0.5h. Add ammonia / methanol solution (67mL) and react at room temperature for 5h. Then add ammonia / methanol solution (67mL) and react at 60℃ overnight. After the reaction is completed, the reaction solution is concentrated, and a large amount of white solid precipitates. Filter with suction, and wash the filter cake with a small amount of methanol to obtain the title product. ESI-MS m / z: 304.8[M+H] + .

[0110] Step 3: Synthesis of 1-(4-bromo-2,6-difluorophenyl)-3-((2-(trimethylsilyl)ethoxy)methyl)dihydropyrimidine-2,4(1H,3H)-dione

[0111]

[0112] 1-(4-bromo-2,6-difluorophenyl)dihydropyrimidine-2,4(1H,3H)-dione (5.0 g, 16.45 mmol) was added to a single-necked flask, and tetrahydrofuran (50 mL) was added to dissolve it, and then ice-bathed to 0°C, and then 1,8-diazabicyclo[5.4.0]undec-7-ene (4.51 g, 29.6 mmol) and 2-(trimethylsilyl)ethoxymethyl chloride (5.49 g, 32.9 mmol) were added, and the reaction was carried out at room temperature for 12 hours. After the reaction was completed, a saturated sodium bicarbonate solution was added to the reaction solution to quench the reaction, and then extracted with ethyl acetate, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product. ESI-MS m / z: 408.9 [M-26] + .

[0113] Step 4: Synthesis of 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-3-((2-(trimethylsilyl)ethoxy)methyl)dihydropyrimidine-2,4(1H,3H)-dione

[0114]

[0115] 1-(4-bromo-2,6-difluorophenyl)-3-((2-(trimethylsilyl)ethoxy)methyl)dihydropyrimidine-2,4(1H,3H)-dione (2.0 g, 4.6 mmol) was added to a single-necked flask, 1,4-dioxane (10 mL) was added to dissolve it, palladium acetate (110 mg, 0.5 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthene (290 mg, 0.5 mmol) and cesium carbonate (4.5 g, 13.8 mmol) were added, and azetidine-3-ol (756 mg, 6.9 mmol) was added, and the reaction was carried out at 100°C for 12 h under inert gas protection. After the reaction was completed, the mixture was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatographed to obtain the title product. ESI-MS m / z: 399.9 [M-28] + .

[0116] Step 5: Synthesis of 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-3-(hydroxymethyl)dihydropyrimidine-2,4(1H,3H)-dione

[0117]

[0118] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-3-((2-(trimethylsilyl)ethoxy)methyl)dihydropyrimidine-2,4(1H,3H)-dione (1.0 g, 2.34 mmol) was added to a single-necked flask, dichloromethane (10 mL) was added to dissolve it, and then trifluoroacetic acid (2 mL) was added to react at room temperature for 1 h. After the reaction was completed, the reaction solution was concentrated to obtain the title product. ESI-MS m / z: 327.9 [M+H] + .

[0119] Step 6: Synthesis of 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0120]

[0121] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-3-(hydroxymethyl)dihydropyrimidine-2,4(1H,3H)-dione (766 mg, 2.34 mmol) was added to a single-necked flask, acetonitrile (5 mL) was added to dissolve it, and then ammonia water (0.5 mL) was added to react at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated and column chromatography was performed to obtain the title product. ESI-MS m / z: 297.9 [M+H] + .

[0122] Step 7: Synthesis of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate

[0123]

[0124] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (110mg, 0.37mmol) was added to a single-necked flask, N,N-dimethylformamide (5mL) was added to dissolve it, and phenyl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate (123mg, 0.39mmol) was added, ice-bathed to 0°C, and sodium hydrogen (18mg, 0.44mmol) was added, and the reaction was carried out at 0°C for 1h. After the reaction was completed, a small amount of dilute hydrochloric acid solution (3mmol / L) was added to the reaction solution to adjust the pH to neutral, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated the organic phase, and column chromatography was performed to obtain the title product. ESI-MS m / z: 518.8[M+H] + .1 H NMR (400 MHz, DMSO-d 6 )δ10.50(s,1H),9.98(s,1H),7.80(s,1H),7.42-7.37(m,1H),7.16(d,J=7.5Hz,1H),6.31(d,J=10.1 Hz,2H),5.30(s,1H),4.28-4.22(m,2H),3.88(d,J=7.1Hz,2H),3.62-3.56(m,2H),2.70-2.66(m,2H).

[0125] Example 2: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(4-chloro-3-(cyclopropylethynyl)phenyl)carbamate

[0126]

[0127] Step 1: Synthesis of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(4-chloro-3-(cyclopropylethynyl)phenyl)carbamate

[0128]

[0129] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (110mg, 0.37mmol) was added to a single-necked flask, N,N-dimethylformamide (5mL) was added to dissolve it, and phenyl(4-chloro-3-(cyclopropylethynyl)phenyl)carbamate (122mg, 0.39mmol) was added, ice-bathed to 0°C, and sodium hydrogen (18mg, 0.44mmol) was added, and the reaction was carried out at 0°C for 1h. After the reaction was completed, a small amount of dilute hydrochloric acid solution (3mmol / L) was added to the reaction solution to adjust the pH to neutral, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated the organic phase, and column chromatographed to obtain the title product. ESI-MS m / z: 514.9[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ10.50(s,1H),10.10(s,1H),7.58(s,1H),7.44-7.36(m,2H),6.30(d,J=10.2Hz,2H),5.31-5.25(m,1H),4.28-4.20(m,2H),3. 86(dd,J=9.0,2.6Hz,2H),3.59(t,J=6.3Hz,2H),2.68(t,J=6.2Hz,2H),1.63-1.57(m,1H),0.97-0.90(m,2H),0.79-0.72(m,2H).

[0130] Example 3: Preparation of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(4-chloro-3-(cyclopropylethynyl)phenyl)carbamate

[0131]

[0132] Step 1: Synthesis of 5-bromo-2-(bromomethyl)-1,3-difluorobenzene

[0133]

[0134] 5-Bromo-1,3-difluoro-2-methylbenzene (25.0 g, 120.76 mmol) was added to a single-necked flask, carbon tetrachloride (250 mL) was added to dissolve it, and then N-bromosuccinimide (21.5 g, 120.76 mmol) and azobisisobutyronitrile (992 mg, 6.04 mmol) were added, and the reaction was carried out at 80°C for 2 h under inert gas protection. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and column chromatography was performed to obtain the title product.

[0135] Step 2: Synthesis of 2-(4-bromo-2,6-difluorophenyl)acetonitrile

[0136]

[0137] 5-Bromo-2-(bromomethyl)-1,3-difluorobenzene (23.0 g, 80.44 mmol) was added to a single-necked flask, acetonitrile (120 mL) was added to dissolve it, and then trimethylsilyl cyanide (16.0 g, 160.88 mmol) and potassium carbonate (22.2 g, 160.88 mmol) were added, and the reaction was carried out at 90°C for 12 hours. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, and column chromatography was performed to obtain the title product.

[0138] Step 3: Synthesis of ethyl 4-(4-bromo-2,6-difluorophenyl)-4-cyanobutyrate

[0139]

[0140] 2-(4-bromo-2,6-difluorophenyl)acetonitrile (11.90 g, 51.3 mmol) was added to a single-necked flask, tetrahydrofuran (110 mL) was added to dissolve it, and then ethyl acrylate (5.13 g, 51.3 mmol) was added, ice-bathed to 0°C, sodium methoxide (2.77 g, 51.3 mmol) was added, and the reaction was allowed to react at room temperature for 1 hour. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, and then ethyl acetate was extracted, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product.

[0141] Step 4: Synthesis of 3-(4-bromo-2,6-difluorophenyl)piperidin-2,6-one

[0142]

[0143] Add 4-(4-bromo-2,6-difluorophenyl)-4-cyanobutyric acid ethyl ester (13.5 g, 40.6 mmol) to a three-necked flask, add acetic acid (25 mL) to dissolve it, then add concentrated sulfuric acid (13.2 mL) and react at 90°C for 2 h. After the reaction is completed, add an appropriate amount of ice water to the reaction solution, a large amount of white solid precipitates, filter, and wash the filter cake with water 2-3 times to obtain the title product. ESI-MS m / z: 303.8 [M+H] + .

[0144] Step 5: Synthesis of 3-(4-bromo-2,6-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-2,6-one

[0145]

[0146] 3-(4-bromo-2,6-difluorophenyl)piperidin-2,6-one (11.0 g, 36.5 mmol) was added to a single-necked flask, and tetrahydrofuran (40 mL) was added to dissolve it, and then ice-bathed to 0°C, and then 1,8-diazabicyclo[5.4.0]undec-7-ene (10.0 g, 65.7 mmol) and 2-(trimethylsilyl)ethoxymethyl chloride (12.2 g, 73.0 mmol) were added, and the reaction was carried out at room temperature for 12 hours. After the reaction was completed, a saturated sodium bicarbonate solution was added to the reaction solution to quench the reaction, and then ethyl acetate was extracted, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product. ESI-MS m / z: 432.8 [MH] - .

[0147] Step 6: Synthesis of 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione

[0148]

[0149] 3-(4-bromo-2,6-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-2,6-one (3.0 g, 6.91 mmol) was added to a single-necked flask, 1,4-dioxane (10 mL) was added to dissolve it, palladium acetate (155 mg, 0.69 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthene (399 mg, 0.69 mmol) and cesium carbonate (6.75 g, 20.73 mmol) were added, and azetidine-3-ol (1.14 g, 10.37 mmol) was added, and the reaction was carried out at 100°C for 12 hours under inert gas protection. After the reaction was completed, the mixture was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to obtain the title product.

[0150] Step 7: Synthesis of 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-1-(hydroxymethyl)piperidine-2,6-dione

[0151]

[0152] 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (1.36 g, 3.19 mmol) was added to a single-necked flask, dichloromethane (15 mL) was added to dissolve it, and then trifluoroacetic acid (3 mL) was added to react at room temperature for 12 hours. After the reaction was completed, the reaction solution was concentrated to obtain the title product. ESI-MS m / z: 326.9 [M+H] + .

[0153] Step 8: Synthesis of 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)piperidine-2,6-dione

[0154]

[0155] 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)-1-(hydroxymethyl)piperidine-2,6-dione (1.04 mg, 3.19 mmol) was added to a single-necked flask, acetonitrile (10 mL) was added to dissolve it, and then ammonia water (1.0 mL) was added to react at room temperature for 1 h. After the reaction was completed, the reaction solution was concentrated and column chromatography was performed to obtain the title product. ESI-MS m / z: 297.0 [M+H] + .

[0156] Step 9: Synthesis of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(4-chloro-3-(cyclopropylethynyl)phenyl)carbamate

[0157]

[0158] 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)piperidine-2,6-dione (200 mg, 0.68 mmol) was added to a single-necked flask, N,N-dimethylformamide (4 mL) was added to dissolve it, and phenyl(4-chloro-3-(cyclopropylethynyl)phenyl)carbamate (221 mg, 0.71 mmol) was added, ice-bathed to 0°C, and sodium hydrogen (33 mg, 0.82 mmol) was added, and the reaction was carried out at 0°C for 1 hour. After the reaction was completed, a small amount of dilute hydrochloric acid solution (3 mmol / L) was added to the reaction solution to adjust the pH to neutral, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated the organic phase, and column chromatographed to obtain the title product. ESI-MS m / z: 513.9 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ10.90-10.83(m,1H),10.09(s,1H),7.58(s,1H),7.43-7.36(m,2H),6.22(d,J =11.2Hz,2H),5.31-5.24(m,1H),4.22(t,J=7.4Hz,2H),4.05(dd,J=12.3,4.2Hz, 1H),3.84(d,J=6.5Hz,2H),2.83-2.73(m,1H),2.57-2.52(m,1H),2.12-2.05(m, 1H),1.98-1.91(m,1H),1.64-1.56(m,1H),0.98-0.90(m,2H),0.80-0.73(m,2H).

[0159] Example 4: Preparation of N-(1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl)-2-oxo-2-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)acetamide

[0160]

[0161] Step 1: Synthesis of tert-butyl (1-(4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl)carbamate

[0162]

[0163] 1-(4-bromo-2,6-difluorophenyl)-3-((2-(trimethylsilyl)ethoxy)methyl)dihydropyrimidine-2,4(1H,3H)-dione (2.5 g, 5.76 mmol) was added to a single-necked flask, 1,4-dioxane (20 mL) was added to dissolve it, palladium acetate (130 mg, 0.58 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthene (336 mg, 0.58 mmol) and cesium carbonate (5.6 g, 17.28 mmol) were added, and tert-butyl azetidin-3-ylcarbamate (2.4 g, 11.52 mmol) was added, and the reaction was carried out at 100°C for 12 h under inert gas protection. After the reaction was completed, the mixture was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and subjected to column chromatography to obtain the title product.

[0164] Step 2: Synthesis of 1-(4-(3-aminoazetidin-1-yl)-2,6-difluorophenyl)-3-(hydroxymethyl)dihydropyrimidine-2,4(1H,3H)-dione

[0165]

[0166] Add tert-butyl (1-(4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl)carbamate (640 mg, 1.22 mmol) to a single-necked flask, add dichloromethane (10 mL) to dissolve it, then add boron trifluoride etherate (519 mg, 3.66 mmol) and react at room temperature for 1 hour. After the reaction is completed, the reaction solution is concentrated to obtain the title product. ESI-MS m / z: 326.9 [M+H] + .

[0167] Step 3: Synthesis of 1-(4-(3-aminoazetidin-1-yl)-2,6-difluorophenyl)dihydropyrimidine-2,4(1H,3H)-dione

[0168]

[0169] 1-(4-(3-aminoazetidin-1-yl)-2,6-difluorophenyl)-3-(hydroxymethyl)dihydropyrimidine-2,4(1H,3H)-dione (398 mg, 1.22 mmol) was added to a single-necked flask, acetonitrile (10 mL) was added to dissolve it, and then ammonia water (10 mL) was added to react at room temperature for 1 h. After the reaction was completed, the reaction solution was concentrated and column chromatography was performed to obtain the title product. ESI-MS m / z: 296.9 [M+H] + .

[0170] Step 4: Synthesis of 1-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)ethan-1-one

[0171]

[0172] 1-Bromo-4-(1-(trifluoromethyl)cyclopropyl)benzene (400 mg, 1.059 mmol) was added to a single-necked flask, 1,4-dioxane (10 mL) was added to dissolve it, and then tributyl(1-ethoxyvinyl)stannane (818 mg, 2.263 mmol) and tetrakistriphenylphosphine palladium (349 mg, 0.301 mmol) were added, and the reaction was carried out at 100°C for 12 hours under inert gas protection. After the reaction was completed, the reaction solution was concentrated and the title product was obtained by column chromatography. ESI-MS m / z: 228.9 [M+H] + .

[0173] Step 5: Synthesis of 2-oxo-2-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)acetic acid

[0174]

[0175] 1-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)ethane-1-one (278 mg, 1.22 mmol) was added to a single-necked flask, pyridine (10 mL) was added to dissolve it, and then selenium dioxide (271 mg, 2.44 mmol) was added and reacted at 90°C for 2 h. After the reaction was completed, the reaction solution was concentrated and the title product was obtained by column chromatography. ESI-MS m / z: 256.0 [MH] - .

[0176] Step 6: Synthesis of 2-oxo-2-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)acetyl chloride

[0177]

[0178] 2-Oxo-2-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)acetic acid (50 mg, 0.194 mmol) was added to a single-necked flask, and dichloromethane (2 mL) was added to dissolve it. Then, oxalyl chloride (49 mg, 0.388 mmol) and 1 drop of N,N-dimethylformamide were added, and the reaction was carried out at room temperature for 1 hour. After the reaction was completed, the reaction solution was concentrated to obtain the title product.

[0179] Step 7: Synthesis of N-(1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl)-2-oxo-2-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)acetamide

[0180]

[0181] 1-(4-(3-aminoazetidin-1-yl)-2,6-difluorophenyl)dihydropyrimidine-2,4(1H,3H)-dione (57 mg, 0.194 mmol) was added to a single-necked flask, and dichloromethane (3 mL) was added to dissolve it. Then triethylamine (39 mg, 0.388 mmol) and 2-oxo-2-(4-(1-(trifluoromethyl)cyclopropyl)phenyl)acetyl chloride (54 mg, 0.194 mmol) were added, and the reaction was carried out at room temperature for 2 hours. After the reaction was completed, the reaction solution was concentrated and column chromatography was performed to obtain the title product. ESI-MS m / z: 537.1[M+H] + . 1 HNMR (400 MHz, DMSO-d 6 )δ10.50(s,1H),9.67(d,J=6.4Hz,1H),8.02(d,J=7.9Hz,2H),7.67(d,J=7.7Hz,2H),6.28(d,J=10.0Hz,2H),4.85-4.70( m,1H),4.33-4.16(m,2H),3.93-3.76(m,2H),3.66-3.53(m,2H),2.77-2.63(m,2H),1.48-1.35(m,2H),1.26-1.16(m,2H).

[0182] Example 5: Preparation of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)-3-methylazetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate

[0183]

[0184] Step 1: Preparation of 3-(2,6-difluoro-4-(3-hydroxy-3-methyl-1-yl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione

[0185]

[0186] 3-(4-bromo-2,6-difluorophenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-2,6-one (2.2 g, 5.07 mmol) was added to 1,4-dioxane (20 mL), followed by palladium acetate (100 mg, 0.45 mmol), 4,5-bis(diphenylphosphino)-9,9-dimethyloxanthene (270 mg, 0.47 mmol) and cesium carbonate (4.5 g, 13.8 mmol), and then azetidine-3-ol (600 mg, 6.89 mmol), inert gas protection, and reaction at 100 ° C for 12 h. After the reaction, the mixture was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography to obtain the title product. ESI-MS m / z: 440.9 [M+H] + .

[0187] Step 2: Preparation of 1-(4-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-3,5-difluorophenyl)-3-methylazetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate

[0188]

[0189] 3-(2,6-difluoro-4-(3-hydroxy-3-methyl-1-yl)phenyl)-1-((2-(trimethylsilyl)ethoxy)methyl)piperidine-2,6-dione (150 mg, 0.34 mmol) was added to N,N-dimethylformamide (5 mL), ice-bathed to 0°C, sodium hydrogen (16 mg, 0.41 mmol) was added, and then phenyl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate (113 mg, 0.36 mmol) was added, and the reaction was carried out at 0°C for 1 hour. After the reaction was completed, a small amount of dilute hydrochloric acid solution (3 mmol / L) was added to the reaction solution to adjust the pH to neutral, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated the organic phase, and column chromatographed to obtain the title product. ESI-MS m / z: 662.0 [M+H] + .

[0190] Step 3: Preparation of 1-(3,5-difluoro-4-(1-(hydroxymethyl)-2,6-dioxopiperidin-3-yl)phenyl)-3-methylazetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl

[0191]

[0192] 1-(4-(2,6-dioxo-1-((2-(trimethylsilyl)ethoxy)methyl)piperidin-3-yl)-3,5-difluorophenyl)-3-methylazetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate (95 mg, 0.14 mmol) was added to dichloromethane (5 mL), and then trifluoroacetic acid (1 mL) was added and reacted at room temperature for 3 h. After the reaction was completed, the reaction solution was concentrated and used directly in the next step. ESI-MS m / z: 562.1[M+H] + .

[0193] Step 4: Preparation of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)-3-methylazetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl)carbamate

[0194]

[0195] 1-(3,5-difluoro-4-(1-(hydroxymethyl)-2,6-dioxopiperidin-3-yl)phenyl)-3-methylazetidin-3-yl(2-fluoro-5-(trifluoromethoxy)phenyl) (79 mg, 0.14 mmol) was added to acetonitrile (10 mL), and then ammonia water (1 mL) was added, and the reaction was carried out at room temperature for 1 h. After the reaction, a small amount of dilute hydrochloric acid solution (3 mmol / L) was added to the reaction solution to adjust the pH to neutral, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated to prepare the title product. ESI-MS m / z: 532.1[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ10.95(s,1H),7.65(d,J=12.0Hz,2H),7.27(t,J=9.3Hz,1H),7.19(d,J=4 .3Hz,1H),7.02-6.95(m,1H),5.40-5.32(m,1H),4.21(dd,J=8.4Hz,1H),4. 08(d,J=9.2Hz,1H),3.87(d,J=9.0Hz,1H),3.55-3.44(m,2H),2.86-2.77(m ,1H),2.56(s,1H),2.15(d,J=10.7Hz,1H),2.05-1.97(m,1H),1.41(s,3H).

[0196] Example 6: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-ylbenzo[d]thiazol-2-ylcarbamate

[0197]

[0198] Step 1: Preparation of 1-(4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-ylphenyl carbonate

[0199]

[0200] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl))phenyl)-3-((2-(trimethylsilyl)ethoxy)methyl)dihydropyrimidine-2,4(1H,3H)-dione (480 mg, 1.12 mmol) was added to a single-necked flask, dichloromethane (8 mL) was added to dissolve it, and then pyridine (266 mg, 3.36 mmol) was added. The mixture was protected by inert gas and ice-bathed to 0°C. Then phenyl chloroformate (877 mg, 5.6 mmol) was added and reacted at room temperature for 1 hour. After the reaction was completed, the mixture was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated and column chromatography was performed to obtain the title product.

[0201] Step 2: Preparation of 1-(4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-ylbenzo[d]thiazol-2-ylcarbamate

[0202]

[0203] 1-(4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-ylphenyl carbonate (200 mg, 0.37 mmol) was added to a single-necked flask, N,N-dimethylformamide (5 mL) was added to dissolve it, and then benzo[d]thiazol-2-amine (62 mg, 0.41 mmol) and cesium carbonate (241 mg, 0.74 mmol) were added, and the reaction was carried out at room temperature for 4 hours. After the reaction was completed, it was extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated and column chromatography was performed to obtain the title product. ESI-MS m / z: 604.1[M+H] + .

[0204] Step 3: Preparation of 1-(3,5-difluoro-4-(3-(hydroxymethyl)-2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)azetidin-3-ylbenzo[d]thiazol-2-ylcarbamate

[0205]

[0206] 1-(4-(2,4-dioxo-3-((2-(trimethylsilyl)ethoxy)methyl)tetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-ylbenzo[d]thiazol-2-ylcarbamate (205 mg, 0.34 mmol) was added to a single-necked flask, dichloromethane (5 mL) was added to dissolve it, and then trifluoroacetic acid (1 mL) was added to react at room temperature for 1 h. After the reaction was completed, the reaction solution was concentrated and used directly in the next step. ESI-MS m / z: 504.1[M+H] + .

[0207] Step 4: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-ylbenzo[d]thiazol-2-ylcarbamate

[0208]

[0209] 1-(3,5-difluoro-4-(3-(hydroxymethyl)-2,4-dioxotetrahydropyrimidin-1(2H)-yl)phenyl)azetidin-3-ylbenzo[d]thiazol-2-ylcarbamate (171 mg, 0.34 mmol) was added to a single-necked flask, acetonitrile (5 mL) was added to dissolve it, and then ammonia water (0.5 mL) was added to react at room temperature for 1 h. After the reaction was completed, the reaction solution was concentrated and column chromatography was performed to prepare the title product. ESI-MS m / z: 474.1[M+H] + . 1H NMR (400 MHz, DMSO-d 6 )δ12.32(s,1H),10.50(s,1H),7.96(d,J=7.4Hz,1H),7.70(d,J=6.8Hz,1H),7.46-7.38(m,1H),7.33-7.24(m,1H),6.3 2(d,J=9.9Hz,2H),5.44-5.34(m,1H),4.35-4.23(m,2H),3.92(d,J=6.1Hz,2H),3.64-3.54(m,2H),2.73-2.64(m,2H).

[0210] Example 7: Preparation of 1-(5-(cyclopropylethynyl)-2-fluorophenyl)-3-(1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl)urea

[0211]

[0212] Step 1: Preparation of 5-(cyclopropylethynyl)-2-fluoroaniline

[0213]

[0214] Add 2-fluoro-5-iodoaniline (1g, 4.2mmol) to a sealed tube, and then add (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (295mg, 0.42mmol), cuprous iodide (160mg, 0.84mmol), triethylamine (850mg, 8.4mmol), cyclopropylacetylene (1.38g, 21.0mmol), and 1,4-dioxane (20mL). Protect with inert gas, heat to 50°C, react for 12h, monitor the reaction results with LCMS, and after the reaction is completed, filter, concentrate under reduced pressure, and perform column chromatography to obtain the title product. ESI-MS m / z: 176.1[M+H] + .

[0215] Step 2: Preparation of phenyl (5-(cyclopropylethynyl)-2-fluorophenyl) carbamate

[0216]

[0217] 5-(Cyclopropylethynyl)-2-fluoroaniline (380 mg, 2.17 mmol) was added to a single-necked flask, DCM (10 mL) was added to dissolve it, sodium bicarbonate (365 mg, 4.34 mmol) was added, and phenyl chloroformate (510 mg, 3.26 mmol) was added dropwise at 0°C. The reaction was allowed to react at room temperature for 1 h, and the reaction was monitored by LCMS. After the reaction was completed, the reaction solution was concentrated and column chromatography was performed to obtain the title product. ESI-MS m / z: 296.1 [M+H] + .

[0218] Step 3: Preparation of 1-(5-(cyclopropylethynyl)-2-fluorophenyl)-3-(1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl)urea

[0219]

[0220] 1-(4-(3-aminoazetidin-1-yl)-2,6-difluorophenyl)dihydropyrimidine-2,4(1H,3H)-dione (0.09 g, 0.304 mmol) was added to a single-necked flask, dimethylacetamide (3 mL) was added to dissolve it, sodium hydride (24 mg, 0.608 mmol) and phenyl(5-(cyclopropylethynyl)-2-fluorophenyl)carbamate (0.09 g, 0.304 mmol) were added at 0°C, and the reaction was allowed to react at room temperature for 0.5 h. The reaction was monitored by LCMS. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product. ESI-MS m / z: 498.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ10.49(s,1H),8.39(s,1H),8.09(d,J=7.4Hz,1H),7.30(d,J=6.4Hz,1H),7.20-7.10(m,1H),6.95(s,1H),6.27(d,J=9.9Hz,2H),4 .60-4.46(m,1H),4.23-4.10(m,2H),3.76-3.53(m,4H),2.74-2.63(m,2H),1.57-1.44(m,1H),0.94-0.80(m,2H),0.77-0.63(m,2H).

[0221] Example 8: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(2-fluoro-5-(prop-1-yn-1-yl)phenyl)carbamate

[0222]

[0223] Step 1: Preparation of 2-fluoro-5-(prop-1-yn-1-yl)aniline

[0224]

[0225] Add 2-fluoro-5-iodoaniline (2.5 g, 10.6 mmol) to a sealed tube, add 1,4-dioxane (10 mL) to dissolve it, then add bistriphenylphosphine palladium dichloride (744 mg, 1.06 mml), cuprous iodide (404 mg, 2.12 mmol) and triethylamine (2.15 g, 21.2 mmol), then add propyne (848 mg, 21.2 mmol), protect with inert gas, react at 45 ° C for 12 hours. After the reaction is completed, the reaction solution is concentrated under reduced pressure and column chromatography is performed to obtain the title product. ESI-MS m / z: 150.1 [M+H] + .

[0226] Step 2: Preparation of phenyl (2-fluoro-5- (prop-1-yn-1-yl) phenyl) carbamate

[0227]

[0228] 2-Fluoro-5-(prop-1-yn-1-yl)aniline (270 mg, 1.81 mmol) was added to a two-necked flask, dissolved in dichloromethane (5 mL), pyridine (287 mg, 3.62 mmol) was added, stirred at 0°C, phenyl chloroformate (312 mg, 1.99 mmol) was slowly added dropwise, and the reaction was carried out at room temperature for 0.5 h. After the reaction was completed, water was added to quench, and then extracted with dichloromethane, washed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product. ESI-MS m / z: 270.0 [M+H] + .

[0229] Step 3: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(2-fluoro-5-(prop-1-yn-1-yl)phenyl)carbamate

[0230]

[0231] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (125 mg, 0.422 mmol) was added to a single-necked flask, dissolved in N,N-dimethylformamide (8 mL), NaH (20 mg, 0.844 mmol) was added at 0°C, and phenyl (2-fluoro-5-(prop-1-yn-1-yl)phenyl)carbamate (125 mg, 0.464 mmol) was added dropwise, and the reaction was carried out at room temperature for 0.5 h. After the reaction was completed, saturated ammonium chloride solution was added to quench, and then extracted with ethyl acetate, washed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product. ESI-MS m / z: 473.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ10.49(s,1H),9.70(s,1H),7.73-7.49(m,1H),7.29-6.97(m,2H),6.44-6.15(m,2H),5.28(s,1H), 4.40-4.14(m,2H),3.86(d,J=6.1Hz,2H),3.59(t,J=6.5Hz,2H),2.68(t,J=6.5Hz,2H),2.02(s,3H).

[0232] Example 9: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(3-(cyclopropylethynyl)-4-(difluoromethoxy)phenyl)carbamate

[0233]

[0234] Step 1: Preparation of 2-bromo-1-(difluoromethoxy)-4-nitrobenzene

[0235]

[0236] Dissolve 2-bromo-4-nitrophenol (2 g, 9.2 mmol) in anhydrous N,N-dimethylformamide (40 mL), add sodium hydrogen sulfide (552 mg, 13.8 mmol) at room temperature, react at room temperature for 1 h, add sodium difluorochloroacetate, heat to 55 °C and stir to react for 16 h, monitor the reaction by TLC, and quench the reaction after the reaction is completed. Extract with ethyl acetate, wash with saturated sodium chloride solution, dry the organic phase over anhydrous sodium sulfate, concentrate under reduced pressure, and perform column chromatography to obtain the title product. 1 H NMR (400 MHz, CDCl 3)δ8.54(d,J=2.1Hz,1H),8.31-8.11(m,1H),7.38(d,J=8.9Hz,1H),6.67(t,J=71.7Hz,1H).

[0237] Step 2: Preparation of 3-bromo-4-(difluoromethoxy)aniline

[0238]

[0239] The compound 2-bromo-1-(difluoromethoxy)-4-nitrobenzene (633mg, 2.36mmol) was dissolved in ethanol (10mL), and iron powder (661mg, 11.8mmol), ammonium chloride (505mg, 9.44mmol), and water (2mL) were added. The temperature was raised to 80°C and stirred for reaction. The reaction was monitored by LCMS. After the reaction was completed, the reaction solution was filtered, the filtrate was concentrated, diluted with water, and then extracted with ethyl acetate, washed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product. ESI-MSm / z: 237.9 [M+H] + .

[0240] Step 3: Preparation of 3-(cyclopropylethynyl)-4-(difluoromethoxy)aniline

[0241]

[0242] Add 3-bromo-4-(difluoromethoxy)aniline (650 mg, 2.73 mmol) to a sealed tube, add 1,4-dioxane (10 mL) to dissolve it, then add (1,1'-bis(diphenylphosphino)ferrocene)palladium dichloride (400 mg, 0.546 mmol), cuprous iodide (104 mg, 0.546 mmol), N,N-diisopropylethylamine (1.6 g, 8.19 mmol), cyclopropylacetylene (902 mg, 13.65 mmol) in sequence, protect with inert gas, react at 120°C for 12 h, monitor the reaction results with LCMS, after the reaction is completed, add water to quench, extract with ethyl acetate, wash with saturated sodium chloride solution, dry the organic phase over anhydrous sodium sulfate, concentrate under reduced pressure, and perform column chromatography to obtain the title product. ESI-MS m / z: 224.0 [M+H] + .

[0243] Step 4: Preparation of phenyl (3-(cyclopropylethynyl)-4-(difluoromethoxy)phenyl)carbamate

[0244]

[0245] 3-(Cyclopropylethynyl)-4-(difluoromethoxy)aniline (450 mg, 2.02 mmol) was added to a single-necked flask, dichloromethane (10 mL) was added to dissolve it, N-diisopropylethylamine (522 mg, 4.04 mmol) was added, and phenyl chloroformate (474 ​​mg, 3.03 mmol) was added dropwise at 0°C. The reaction was allowed to react at room temperature for 1 h, and the reaction was monitored by LCMS. After the reaction was completed, the reaction solution was concentrated and subjected to column chromatography to obtain the title product. ESI-MS m / z: 344.0 [M+H] + .

[0246] Step 5: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(3-(cyclopropylethynyl)-4-(difluoromethoxy)phenyl)carbamate

[0247]

[0248] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (0.1 g, 0.29 mmol) was added to a single-necked flask, dimethylacetamide (3 mL) was added to dissolve it, sodium hydrogen (25 mg, 0.58 mmol) and phenyl (3-(cyclopropylethynyl)-4-(difluoromethoxy)phenyl)carbamate (0.085 g, 0.29 mmol) were added at 0°C, and the reaction was allowed to react at room temperature for 0.5 h. The reaction was monitored by LCMS. After the reaction was completed, a saturated ammonium chloride solution was added to quench the reaction, and the mixture was extracted with ethyl acetate, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, and subjected to column chromatography to obtain the title product. ESI-MS m / z: 547.1 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ10.50(s,1H),10.02(s,1H),7.52(s,1H),7.42(d,J=8.8Hz,1H),7.16 (d,J=8.8Hz,1H),7.07(t,J=76.0Hz,1H),6.30(d,J=10.0Hz,2H),5.35-5 .22(m,1H),4.32-4.18(m,2H),3.95-3.81(m,2H),3.67-3.52(m,2H),2.7 7-2.63(m,2H),1.63-1.49(m,1H),0.99-0.83(m,2H),0.80-0.63(m,2H).

[0249] Example 10: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(3-chloro-4-(cyclopropylethynyl)phenyl)carbamate

[0250]

[0251] Step 1: Preparation of phenyl (3-chloro-4-(cyclopropylethynyl)phenyl)carbamate

[0252]

[0253] Dissolve 3-chloro-4-(cyclopropylethynyl)aniline (300 mg, 1.57 mmol) in dichloromethane (5 mL), add N,N-diisopropylethylamine (0.545 mL, 3.13 mmol), add phenyl chloroformate (0.295 mL, 2.35 mmol) at room temperature, and react at room temperature for 1 hour. After the reaction is completed, add water to quench, extract with dichloromethane, wash with saturated sodium chloride solution, dry with anhydrous sodium sulfate, concentrate the organic phase, and column chromatography to prepare the title product. ESI-MS m / z: 312.1 [M+H] + .

[0254] Step 2: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(3-chloro-4-(cyclopropylethynyl)phenyl)carbamate

[0255]

[0256] 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (120 mg, 0.41 mmol) was added to a single-necked flask, N,N-dimethylformamide (5 mL) was added to dissolve it, and phenyl(3-chloro-4-(cyclopropylethynyl)phenyl)carbamate (134 mg, 0.43 mmol) was added, ice-bathed to 0°C, and sodium hydrogen (20 mg, 0.49 mmol) was added, and the reaction was carried out at 0°C for 0.5 h. After the reaction was completed, the reaction solution was added with saturated ammonium chloride aqueous solution to quench the reaction, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, and the organic phase was concentrated and column chromatography was performed to prepare the title product. ESI-MS m / z: 515.1[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ10.50(s,1H),10.24(s,1H),7.65(s,1H),7.41(d,J=8.4Hz,1H),7.34(d,J=8.1Hz,1H),6.30(d,J=10.0Hz,2H),5.30(s,1H),4. 25(t,2H),3.90-3.83(m,2H),3.59(t,J=6.3Hz,2H),2.72-2.65(m,2H),1.60-1.53(m,1H),0.94-0.87(m,2H),0.76-0.69(m,2H).

[0257] Example 11: Preparation of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(3-chloro-4-(cyclopropylethynyl)phenyl)carbamate

[0258]

[0259] Step 1: Preparation of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(3-chloro-4-(cyclopropylethynyl)phenyl)carbamate

[0260]

[0261] 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)piperidine-2,6-dione (90 mg, 0.303 mmol) was dissolved in N,N-dimethylformamide (8 mL), sodium hydride (15 mg, 0.364 mmol) was added at 0°C, the reaction was allowed to react for 10 minutes, phenyl(3-chloro-4-(cyclopropylethynyl)phenyl)carbamate (100 mg, 0.318 mmol) was added, and the reaction was allowed to react at 0°C for 1 hour. After the reaction was completed, a saturated aqueous ammonium chloride solution was added to the reaction solution to quench the reaction, the reaction was extracted with ethyl acetate, washed with a saturated sodium chloride solution, dried over anhydrous sodium sulfate, the organic phase was concentrated, and column chromatography was performed to prepare the title product. ESI-MS m / z: 514.1[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ10.86(s,1H),10.23(s,1H),7.65(s,1H),7.40(d,J=8.4Hz,1H),7.34(d,J=8.5H z,1H),6.22(d,J=11.3Hz,2H),5.29(s,1H),4.26-4.17(m,2H),4.04(dd,J=12.3,4. 5Hz,1H),3.84(d,J=6.3Hz,2H),2.80-2.72(m,1H),2.53-2.50(m,1H),2.13-2.03( m,1H),1.99-1.92(m,1H),1.60-1.53(m,1H),0.94-0.87(m,2H),0.77-0.68(m,2H).

[0262] Example 12: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(5-(cyclopropylethynyl)-2-fluorophenyl)carbamate

[0263]

[0264] Step 1: Synthesis of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(5-(cyclopropylethynyl)-2-fluorophenyl)carbamate

[0265]

[0266] At room temperature, 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (100mg, 0.34mmol) was added to a 25ml single-mouth bottle, and N,N-dimethylacetamide (5ml) was added. Sodium hydride (27mg, 0.67mmol) and phenyl (5-(cyclopropylethynyl)-2-fluorophenyl)carbamate (99mg, 0.34mmol) were added under ice bath, and stirred at room temperature for 0.5h. After the reaction was completed, saturated ammonium chloride solution was used to adjust pH=6, ethyl acetate was used for extraction, saturated sodium chloride solution was used for washing, anhydrous sodium sulfate was used for drying, the organic phase was concentrated, and column chromatography was performed to obtain the title product. ESI-MS m / z: 499.1[M+H] + . 1 H NMR (400 MHz, DMSO-d 6)δ10.50(s,1H),9.70(s,1H),7.69-7.58(m,1H),7.26-7.09(m,2H),6.38-6.24(m,2H),5.34-5.22(m,1H),4.31-4.16(m ,2H),3.92-3.78(m,2H),3.61-3.58(m,2H),2.70-2.67(m,2H),1.58-1.46(m,1H),0.92-0.80(m,2H),0.78-0.63(m,2H).

[0267] Example 13: Preparation of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(3-(cyclopropylethynyl)-4-fluorophenyl)carbamate

[0268]

[0269] Step 1: Synthesis of 3-(cyclopropylethynyl)-4-fluoroaniline

[0270]

[0271] Add 3-bromo-4-fluoroaniline (1g, 5.26mmol) and 1,4-dioxane (10mL) to a sealed tube, then add bistriphenylphosphine palladium dichloride (370mg, 0.526mml), cuprous iodide (201mg, 1.05mmol), triethylamine (1.06g, 10.53mmol), cyclopropylacetylene (1.74g, 26.31mmol), inert gas protection, react at 80℃ for 12h. After the reaction is completed, the reaction solution is concentrated under reduced pressure and column chromatography is performed to obtain the title product. ESI-MS m / z: 176.1[M+H] + .

[0272] Step 2: Synthesis of phenyl (3-(cyclopropylethynyl)-4-fluorophenyl) carbamate

[0273]

[0274] Dissolve 3-(cyclopropylethynyl)-4-fluoroaniline (160 mg, 0.91 mmol) in dichloromethane (5 mL), add pyridine (144 mg, 1.83 mmol), stir at 0 °C, slowly add phenyl chloroformate (172 mg, 1.1 mmol) dropwise, and react for 0.5 h. After the reaction is completed, add water to quench, extract with dichloromethane, wash with saturated sodium chloride solution, dry the organic phase over anhydrous sodium sulfate, concentrate under reduced pressure, and obtain the title product by column chromatography. ESI-MS m / z: 296.1 [M+H] +

[0275] Step 3: Synthesis of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(3-(cyclopropylethynyl)-4-fluorophenyl)carbamate

[0276]

[0277] 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)piperidine-2,6-dione (100 mg, 0.34 mmol) was dissolved in N,N-dimethylformamide (4 mL), sodium hydride (15 mg, 0.62 mmol) was added at 0°C, and phenyl (3-(cyclopropylethynyl)-4-fluorophenyl)carbamate (91 mg, 0.31 mmol) was added dropwise, and the reaction was carried out at room temperature for 0.5 h. After the reaction was completed, saturated ammonium chloride solution was added to quench, and then extracted with ethyl acetate, washed with saturated sodium chloride solution, and the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain the title product. ESI-MS m / z: 498.1[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ10.86(s,1H),9.98(s,1H),7.56-7.47(m,1H),7.45-7.34(m,1H),7.20-7 .16(m,1H),6.22(d,J=11.1Hz,2H),5.43-5.16(m,1H),4.28-4.19(m,2H),4 .09-4.01(m,1H),3.85-3.81(m,2H),2.88-2.69(m,1H),2.53-2.51(m,1H), 2.12-1.96(m,2H),1.63-1.52(m,1H),0.97-0.85(m,2H),0.81-0.68(m,2H).

[0278] Example 14: Preparation of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(5-(cyclopropylethynyl)-2-fluorophenyl)carbamate

[0279]

[0280] Step 1: Synthesis of 1-(4-(2,6-dioxopiperidin-3-yl)-3,5-difluorophenyl)azetidin-3-yl(5-(cyclopropylethynyl)-2-fluorophenyl)carbamate

[0281]

[0282] At room temperature, 3-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)piperidine-2,6-dione (160mg, 0.54mmol) was added to a 50ml single-mouth bottle, and N,N-dimethylacetamide (10ml) was added. Sodium hydride (43mg, 0.61mmol) and phenyl (5-(cyclopropylethynyl)-2-fluorophenyl)carbamate (160mg, 0.54mmol) were added under ice bath, and stirred at room temperature for 0.5h. After the reaction was completed, saturated ammonium chloride solution was added to quench the reaction, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and column chromatography was performed to obtain the title product. ESI-MS m / z: 498.2[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ10.86(s,1H),9.70(s,1H),7.64(d,1H),7.26-7.06(m,2H),6.24(s,1H) ,6.21(s,1H),5.34-5.20(m,1H),4.30-4.14(m,2H),4.11-3.98(m,1H),3.9 0-3.74(m,2H),2.85-2.71(m,1H),2.59-2.52(m,1H),2.15-2.01(m,1H),2 .00-1.87(m,1H),1.58-1.45(m,1H),0.94-0.81(m,2H),0.78-0.64(m,2H).

[0283] Example 15: Preparation of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(3-(cyclopropylethynyl)-4-fluorophenyl)carbamate

[0284]

[0285] Step 1: Synthesis of 1-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3,5-difluorophenyl)azetidin-3-yl(3-(cyclopropylethynyl)-4-fluorophenyl)carbamate

[0286]

[0287] At room temperature, 1-(2,6-difluoro-4-(3-hydroxyazetidin-1-yl)phenyl)dihydropyrimidine-2,4(1H,3H)-dione (130mg, 0.44mmol) was added to a 50ml single-mouth bottle, N,N-dimethylacetamide (5ml) was added, sodium hydride (35mg, 0.87mmol) and phenyl(3-(cyclopropylethynyl)-4-fluorophenyl)carbamate (130mg, 0.44mmol) were added under ice bath, and stirred at room temperature for 0.5h. After the reaction was completed, the reaction solution was added to saturated ammonium chloride aqueous solution to quench the reaction, extracted with ethyl acetate, washed with saturated sodium chloride solution, dried over anhydrous sodium sulfate, concentrated the organic phase, and column chromatography was performed to prepare the title product. ESI-MS m / z: 499.2[M+H] + . 1 H NMR (400 MHz, DMSO-d 6 )δ10.50(s,1H),9.99(s,1H),7.51(s,1H),7.38(s,1H),7.21-7.16(m,1H),6.32(s,1H),6.29(s,1H),5.31-5.25(m,1H),4.30- 4.17(m,2H),3.92-3.79(m,2H),3.61-3.58(m,2H),2.67-2.60(m,2H),1.64-1.54(m,1H),0.98-0.85(m,2H),0.81-0.69(m,2H).

[0288] Experimental Example 1: Evaluation of in vitro cell activity of compounds

[0289] 1. Experimental Materials

[0290] Test compounds: the compounds of the present invention prepared in the above examples, each compound was formulated into a 10 mM stock solution with DMSO and finally diluted to 10 concentrations for detection. The final concentrations of the compounds in the NCI-H1155 cell experiment were 20000 nM, 4000 nM, 800 nM, 160 nM, 32 nM, 6.4 nM, 1.28 nM, 0.26 nM, 0.051 nM / 30000 nM, 7500 nM, 1875 nM, 468.75 nM, 117.19 nM, 29.30 nM, 7.32 nM, 1.83 nM, 0.46 nM, and 0.11 nM.

[0291] Human lung cancer cells NCI-H1155 cells (highly expressing MYC): ATCC.

[0292] Reagent: TrypLE TMExpress Enzyme (1X), no phenol red, the product number is Gibco 12604-021; Celltiter Glo assay kit (CTG), the product number is Promega G7573.

[0293] 2. Experimental Methods

[0294] 2.1 Cell culture and passaging

[0295] (1) All cells were cultured according to the method recommended by ATCC, and the cells were cultured during the exponential growth phase.

[0296] (2) Cell culture medium:

[0297] NCI-H1155 cells: mulated Leibovitz's L-15 medium, 10% FBS, 1% penicillin-streptomycin.

[0298] (3) Cell culture conditions: 37°C, 5% CO 2 , 95% air

[0299] (4) Depending on the growth of the cells, the culture medium should be changed every 2-3 days or the cells should be subcultured.

[0300] 2.2 Cell plating

[0301] Remove cells from the cell culture flask and resuspend in fresh culture medium. Inoculate 40 μL of the cell resuspension into a 384-well culture plate.

[0302] 2.3 Administration

[0303] NCI-H1155 cells were cultured in the original medium (40 μL). 40 nL of drugs of different concentrations were added to the drug group and 40 nL of DMSO was added to the DMSO group. Two replicate wells were set for each concentration group. 5% CO 2 Incubate for 3 days. The compound was prepared as follows: 1-2 mg of the compound was weighed in advance and prepared into a 10 mM stock solution using DMSO. The drug was diluted with DMSO, with a starting concentration of 10 mM, and then diluted to 10 concentration gradients in a 1:4 gradient. The final concentrations were: 30000 nM, 7500 nM, 1875 nM, 468.75 nM, 117.19 nM, 29.30 nM, 7.32 nM, 1.83 nM, 0.46 nM, and 0.11 nM.

[0304] 2.4 Detection

[0305] After 3 days of drug treatment, the Celltiter Glo assay kit was taken out 30 minutes in advance and equilibrated to room temperature. Then 40 μL of Celltiter-Glo reagent was added to the PC well, the dosing well, and the DMSO well, and shaken for 2 minutes. After incubation at room temperature in the dark for 28 minutes, the luminescent signal was detected. According to the LUM value, the inhibition rate of each well relative to the solvent control well was calculated: Inhibition% = (Ave_H-Sample) / (Ave_H-Ave_L) X 100, Note: H = Ave (DMSO), L = Ave (Medium). According to different drug concentrations and their corresponding inhibition rates, IC was performed using GraghPad 5.0 software. 50 Draw curves, analyze data, and obtain final IC 50 The experimental results are shown in Table 1.

[0306] Table 1 Inhibition of tumor cell growth by compounds

[0307]

[0308] The experimental results show that the compound of the present invention has a strong inhibitory effect on human lung cancer cell NCI-H1155 cells (highly expressing MYC).

[0309] Experimental Example 2: Human Cereblon (CRBN) Binding Experiment (HTRF Method)

[0310] 1. Experimental Materials

[0311] Test compounds: the compounds of the present invention prepared in the above examples, each compound was prepared into a 10 mM stock solution with DMSO, and finally diluted to 10 concentrations for detection, with the final concentrations of the compounds being 100000 nM, 25000 nM, 6250 nM, 1562.5 nM, 390.62 nM, 97.65 nM, 24.41 nM, 6.10 nM, 1.52 nM, and 0.38 nM.

[0312] Reagents: HTRF HUMAN CEREBLON BINDING KITS, Perkinelmer 64BDCRBNPEG

[0313] 2. Experimental Methods

[0314] (1) 0.16 μL of compound solution was transferred to a 384-well reaction plate. The highest experimental concentration was 100 μM. The final concentrations were 100,000 nM, 25,000 nM, 6,250 nM, 1,562.5 nM, 390.62 nM, 97.65 nM, 24.41 nM, 6.10 nM, 1.52 nM, and 0.38 nM after 4-fold dilution.

[0315] (2) Add 8 μL of 2× human cereblon protein to the 384 reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at 25°C for 15 min.

[0316] (3) Place the mixture of thalidomide red and GST EU in a 384 reaction plate, centrifuge at 1000 rpm for 1 min, and incubate at 25°C for 180 min.

[0317] (4) The HTRF signal (Ratio 665 / 615 nm) was read using a BMG microplate reader, and IC was analyzed using GraphicPad 5.0 software according to different drug concentrations and their corresponding inhibition rates. 50 Draw curves, analyze data, and obtain final IC 50 value.

[0318] The IC50 value of the test compound indicates the ability to bind to the CRBN protein, and the lower the value, the stronger the binding ability.

[0319] Experimental Example 3: Evaluation of the ability of compounds to degrade GSPT1 protein

[0320] 1. Experimental Materials

[0321] Test compounds: The compounds of the present invention prepared in the above examples, each compound was prepared into a 10 mM stock solution with DMSO, and finally diluted to 9 concentrations for detection. The final concentrations of the compounds in the HEK293-GSPT1-HiBiT reporter assay were 1000 nM, 333.333 nM, 111.111 nM, 37.037 nM, 12.346 nM, 4.115 nM, 1.372 nM, 0.457 nM, and 0.152 nM.

[0322] Cell line: HEK293 GSPT1 HiBiT KI cells (Beijing Aisiyipu Technology Co., Ltd.).

[0323] Reagents: HiBiT Lytic Detection System (Promega, Catalog No.: N3040).

[0324] 2. Experimental Methods

[0325] (1) Experimental procedures

[0326] HEK293 GSPT1 HiBiT KI cells were cultured in DMEM medium containing 10% FBS and 1% dual antibody. When the cell density reached 80-90%, the cells were digested with trypsin and resuspended into single cells. The digested cells were then plated at 1×104 Add 40 μL per well to a 384-well plate, and place it in a 37°C carbon dioxide incubator for further culture. After the cells attach to the wall, add the compound, starting at 1 μM, dilute 3 times, and continue to culture in a 37°C carbon dioxide incubator for 24 hours. Return Nano-Glo HiBiT to room temperature, then dilute LgBiT protein 1:100 and Nano-Glo HiBiT cleavage substrate 1:2 into an appropriate volume of Nano-Glo HiBiT cleavage buffer, add a new EP tube, and flip and mix evenly. Equilibrate the cell culture plate at room temperature for 10 minutes. Add 40 μL Nano-Glo HiBiT cleavage buffer to the plate and shake it at 300 rpm for 3 minutes in the dark. Read the luminescence value with BMG PHERAstar FSX.

[0327] (2) Data processing

[0328] Calculate the DC50 of a compound using the following formula:

[0329] Y=Bottom+(Top-Bottom) / (1+10^((LogDC50-X)x HillSlope))

[0330] X: cpd concentration; Y: %Protein remain; Top and Bottom: Plateaus insame units as Y;

[0331] logDC50: same log units as X; HillSlope: Slope factor or Hill slope.

[0332] DC50: half degradation concentration; Dmax: maximum degradation level.

[0333] 3. Experimental results

[0334] Table 2 Degradation of GSPT1 (eRF3) by compounds

[0335] Compound <![CDATA[DC 50 (nM)]]> Dmax(%) Example 1 9.6 93.33 Example 2 21.25 91.05 Example 6 9.53 93.24 Example 8 5.24 93.10 Example 9 10.04 91.80 Example 10 30.17 89.86 Embodiment 11 3.6 93.56 Example 12 15.97 91.97 Embodiment 13 1.96 92.89

[0336] The experimental results show that the compound of the present invention exhibits excellent protein degradation ability in the GSPT1 (eRF3) protein degradation experiment.

[0337] Although the present invention has been described in detail above, it is understood by those skilled in the art that various modifications and changes can be made to the present invention without departing from the spirit and scope of the present invention. The scope of the present invention is not limited to the detailed description above, but should be attributed to the claims.

Claims

1. A compound represented by general formula (I) or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, in, X1 is selected from -CH- and -N-; R 1 , R 2 , R 3 , R 4 are each independently selected from hydrogen, halogen, alkyl and haloalkyl; R 5 is selected from hydrogen and alkyl; R 6 is selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, cycloalkyl and heterocyclyl; which is optionally substituted with halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, cycloalkyl, heterocycloalkyl, aryl and heteroaryl; L1, L2 are each independently selected from a bond, -CH2-, -O-, -NH- and -C(O)-; L3 is selected from a bond and -C≡C-; Cy is selected from aryl and heteroaryl; Y is selected from the group consisting of hydrogen, halogen, hydroxy, alkyl, alkenyl, alkynyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, nitro, carboxyl, cyano, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, alkylaminoacyl, dialkylamino, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl, optionally substituted with halogen, hydroxy, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, hydroxyalkoxy, amino, monoalkylamino, alkylacylamino, alkylacyl, aminoacyl, and alkylaminoacyl; n is selected from 0, 1, 2, 3, 4 and 5; and When X1 is selected from -CH-, L3 is selected from -C≡C-.

2. The compound according to claim 1 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, Where R 1 , R 2 , R 3 , R 4 are each independently selected from hydrogen, halogen, C 1-6 Alkyl and halogenated C 1-6 Alkyl; R 5 Selected from hydrogen, C 1-6 alkyl.

3. The compound according to claim 1 or 2, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, Wherein L1 and L2 are each independently selected from -CH2-, -O-, -NH- and -C(O)-; preferably, L1 is selected from -O- and -NH-, and L2 is selected from -O-, -NH- and -C(O)-.

4. The compound according to any one of claims 1 to 3, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Cy is selected from C 6-8 Aryl and 5-10 membered heteroaryl; R 6 Selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halo C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino, C 3-8 Cycloalkyl and 3-8 membered heterocyclic groups, which are optionally substituted by halogen, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, C 3-8 Cycloalkyl, C 3-8 Heterocycloalkyl, C 6-12 Aryl and C 5-12 Heteroaryl substituted; Y is selected from halogen, hydroxyl, C 1-6 Alkyl, C 2-6 Alkenyl, C 2-6 Alkynyl, halo C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, nitro, carboxyl, cyano, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl, C 1-6 Alkylaminoacyl, di-C 1-6 Alkylamino, C 3-8 Cycloalkyl, C 3-8 Heterocycloalkyl, C 6-12 Aryl and C 5-12 Heteroaryl, which is optionally substituted by halogen, hydroxy, C 1-6 Alkyl, halogenated C 1-6 Alkyl, hydroxyl C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, hydroxyl C 1-6 Alkoxy, amino, mono C 1-6 Alkylamino, C 1-6 Alkyl acylamino, C 1-6 Alkyl acyl, amino acyl and C 1-6 Alkylaminoacyl substituted; n is selected from 0, 1, 2 and 3.

5. The compound according to any one of claims 1 to 4, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein the general formula (I) has the structure of the following general formula (Ia), in, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , L1, L2, L3, Y and n have the definitions described in the above general formula (I), preferably, R 6 is selected from fluorine, chlorine, bromine, iodine, hydroxy, methyl, ethyl, propyl, isopropyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, C 3-6 Cycloalkyl, 3-6 membered heterocyclic group, Y is selected from methyl, ethyl, propyl, isopropyl, It is optionally replaced by one or more hydrogen, halogen, hydroxyl, C 1-3 Alkyl, halogenated C 1-3 Alkyl, hydroxyl C 1-3 Alkyl, C 1-3 Alkoxy, halogenated C 1-3 Alkoxy, hydroxyl C 1-3 Alkoxy, amino, mono C 1-3 Alkylamino, C 1-3 Alkyl acylamino, C 1-3 Alkyl acyl, amino acyl and C 1-3 Alkylaminoacyl substitution.

6. The compound according to claim 5 or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein Selected from 7. The compound according to any one of claims 1 to 4, or its isomer, pharmaceutically acceptable salt, solvate, crystal or prodrug, wherein the general formula (I) has the structure of the following general formula (Ib), in, X1, R 1 , R 2 , R 3 , R 4 , R 5 , L1, L2, R 6 , Y and n have the same meanings as described above for the general formula (I).

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

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

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

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