[go: up one dir, main page]

WO2024222918A1 - 一种苯并六元杂环类gspt1蛋白降解剂及其应用 - Google Patents

一种苯并六元杂环类gspt1蛋白降解剂及其应用 Download PDF

Info

Publication number
WO2024222918A1
WO2024222918A1 PCT/CN2024/090223 CN2024090223W WO2024222918A1 WO 2024222918 A1 WO2024222918 A1 WO 2024222918A1 CN 2024090223 W CN2024090223 W CN 2024090223W WO 2024222918 A1 WO2024222918 A1 WO 2024222918A1
Authority
WO
WIPO (PCT)
Prior art keywords
alkyl
cancer
compound
alkynyl
alkenyl
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/CN2024/090223
Other languages
English (en)
French (fr)
Inventor
李志裕
徐熙
常秀锦
卞金磊
瞿凡贵
王举波
范忠蓬
邱志霞
吴红茜
张迪
张艳青
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
China Pharmaceutical University
Ascentage Pharma Suzhou Co Ltd
Original Assignee
China Pharmaceutical University
Ascentage Pharma Suzhou Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by China Pharmaceutical University, Ascentage Pharma Suzhou Co Ltd filed Critical China Pharmaceutical University
Publication of WO2024222918A1 publication Critical patent/WO2024222918A1/zh
Anticipated expiration legal-status Critical
Pending legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A61P35/02—Antineoplastic agents specific for leukemia
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/04—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings directly linked by a ring-member-to-ring-member bond

Definitions

  • the present invention belongs to the field of medical technology, and specifically relates to a benzo six-membered heterocyclic GSPT1 protein degradation agent and a preparation method and application thereof.
  • Targeted protein degradation technology is a new technology that interferes with protein function by utilizing the protein degradation mechanism inherent in eukaryotic cells to regulate protein homeostasis.
  • TPD currently mainly degrades target proteins through ubiquitin proteasomes and lysosomes.
  • PROTAC proteolysis-targeting chimeras
  • MGs molecular glues
  • Molecular glues are small molecules that induce proximity and can provide precise temporal control of various biological processes, such as signal transduction, transcription, chromatin regulation, and protein folding, localization, and degradation. As chemical inducers of proximity, they can induce protein-protein interactions between E3 ubiquitin ligases and target proteins, leading to target protein degradation. Molecular glues can promote dimerization or colocalization of two proteins by forming a ternary complex, resulting in a variety of biological and pharmacological functions.
  • G1 to S phase transition 1 (GSPT1, also known as eRF3a) is a translation termination factor in the body. Downregulation of GSPT1 can lead to abnormal expression of key proteins and inhibit proliferation or induce apoptosis in various tumor cells. Since GSPT1 was found to be a new substrate of E3 ubiquitin ligase CRBN, it can be targeted for degradation by molecular glue degraders. Currently, a variety of molecular glue degraders are in clinical and preclinical studies. These molecules are very effective in treating blood tumors and certain solid tumors. GSPT1 is multifunctional.
  • GSPT1 In addition to being a second-class peptide chain release factor and participating in protein translation termination, it is also related to apoptosis, cell cycle regulation, tumor occurrence and development, and other processes.
  • GSPT1 is processed by proteolysis to form another isoform, which contains a conserved N-terminal apoptosis inhibitor protein binder.
  • IAPs inhibitor of apoptosis proteins
  • the M domain of eRF3 interacts with Polya-binding protein (PABP) protein, coupling translation termination and mRNA degradation.
  • PABP Polya-binding protein
  • AML Acute myeloid leukemia
  • GSPT1 as a new substrate of CRL4-CRBN-E3 ubiquitin ligase, plays an important role in the treatment of AML.
  • CC-90009 is mainly used to treat AML by intravenous injection, while BTX-1188 is an oral drug used to treat AML and NHL.
  • GSPT1 degradation of GSPT1 is associated with the upregulation of ATF3 and ATF4 gene expression, and ATF3 and ATF4 genes are very important for the integrated stress response pathway.
  • the activation of the integrated stress response pathway is closely related to the phosphorylation of eRF2. When the integrated stress response pathway is activated, it will lead to acute apoptosis of cells.
  • breast cancer is the leading cause of death from malignant tumors in women.
  • Wang et al. analyzed the gene expression in the total RNA extracted from the breast cancer samples they collected and identified five genes, including GSPT1, that can be used as potential therapeutic targets for triple-negative breast cancer.
  • GSPT1 genes that can be used as potential therapeutic targets for triple-negative breast cancer.
  • the study by Malta-Vacas et al. also showed that the longer GSPT1 allele 12-GGC is present in 5.1% of breast cancer patients.
  • mRNA quantitative experiments also showed that GSPT1 is overexpressed in tumor tissues of patients with the 12-GGC allele relative to normal adjacent tissues.
  • Miri et al. conducted a test on the GSPT1 gene and breast cancer susceptibility in breast cancer patients.
  • GSPT1 GSPT1-induced GSPT1
  • GSPT1 is also involved in cytoskeleton formation and controlling chromosome separation, the effect of GSPT1 on chromosome separation and cytoplasmic division may also be a potential mechanism for the gene to affect gastric cancer.
  • Liver cancer is a common tumor disease. According to clinical data, the expression of GSPT1 in liver cancer tissues is generally higher than that in normal cells. Since one of the main functions of GSPT1 is to regulate the transition of cells from the G1 phase to the S phase, some researchers have speculated that interfering with the mRNA expression of GSPT1 can change the cell cycle, thereby increasing the percentage of cells in the G1 phase, leading to a decrease in the vitality and proliferation of tumor cells. Although GSPT1 has a carcinogenic effect in the occurrence and development of liver cancer, some researchers have found that GSPT1 may inhibit the carcinogenic process in the development of liver cancer by constructing HepG2 cells that overexpress and knock down the GSTP1 gene.
  • Colorectal cancer is one of the most important malignant tumors of the digestive system.
  • Xiao et al.'s study showed that GSPT18 was overexpressed in human colorectal cancer HCT116 cells compared with colorectal cells in the normal control group. Further experiments also showed that knocking down GSPT1 in colorectal cancer cells with high GSPT1 expression could inhibit the proliferation and migration of colorectal cancer cells. Research results showed that GSPT1 inhibited the cell cycle progression of HCT116 cells through the mTOR pathway. When the mTOR pathway was activated, it would lead to increased translation of mRNA subgroups. Therefore, GSPT1 may play the role of a proto-oncogene in the occurrence and development of colorectal cancer.
  • GSPT1 plays an important role in the occurrence and development of related solid tumors. However, due to its diverse functions and complex regulatory mechanisms, it plays different roles in different tumors. However, it is generally recognized that GSPT1 is a proto-oncogene in most tumors, and in certain specific types of tumors, it plays the role of a tumor suppressor gene. At present, there is still a lack of in-depth research on the relevant mechanism of action of GSPT1 in tumors, but there is no doubt that GSPT1 has become a powerful potential target in tumor treatment.
  • One of the objectives of the present invention is to provide a benzo six-membered heterocyclic compound of general formula I or a pharmaceutically acceptable salt, tautomer, mesomorph, racemate, enantiomer, diastereomer thereof:
  • X is selected from -CH2- , -NH-, -O-, -S- or -Se-;
  • Y is selected from carbon or nitrogen
  • Z is selected from -CH2- , -CD2- , -C(O)- or -C(S)-;
  • each R 1 is independently selected from hydrogen, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O) 2 R a , or a 3-10 membered cycloalkyl, heterocyclyl, aryl and heteroaryl group containing 0-3 heteroatoms, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl,
  • each R2 is independently selected from hydrogen, C1 - C6 alkyl, haloC1- C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, cyano, nitro, -C(O) NRaRb , -C(O) Ra , -C(O ) ORa , -ORa, -OC(O) Ra , -OC (O) ORa , -OC ( O ) NRaRb, -NRaRb , -SRa , -S(O) Ra , -S(O) 2Ra , or 3-10 membered cycloalkyl, heterocyclyl , aryl and heteroaryl containing 0-3 heteroatoms, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are unsubstituted or optionally substituted with
  • each R3 is independently selected from hydrogen, C1 - C6 alkyl, haloC1-C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, cyano, nitro, -C(O) NRaRb , -C(O) Ra , -C(O ) ORa , -ORa, -OC(O) Ra , -OC (O) ORa , -OC ( O ) NRaRb, -NRaRb , -SRa , -S(O) Ra , -S(O) 2Ra , or 3-10 membered cycloalkyl, heterocyclyl , aryl and heteroaryl containing 0-3 heteroatoms, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are unsubstituted or optionally substituted with
  • R4 is selected from hydrogen, deuterium, C1 - C12 alkyl
  • R5 , R6 , R7 , R8 , R9 , and R10 are each independently selected from hydrogen, C1 - C6 alkyl, halogenated C1- C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, cyano, nitro, -C(O) NRaRb , -C(O) Ra , -C(O) ORa , -ORa , -OC (O) Ra , -OC (O) ORa , -OC(O) NRaRb , -NRaRb , -SRa , -S (O )Ra, -S(O)2Ra , or 3-10 membered cycloalkyl, heterocyclyl, aryl and heteroaryl containing 0-3 heteroatoms, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycly
  • Ra and Rb are each independently selected from hydrogen, C1 - C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, hydroxyl, cyano, nitro, benzyl, -C(O) NRcRd , -C(O) Rc , -C(O) ORc , -ORc , -OC(O) Rc , -OC(O) ORc , -OC(O) NRcRd , -NRcRd , -SRc , -S(O ) Rc , -S(O) 2Rc , or 3-10 membered cycloalkyl, heterocyclyl , aryl and heteroaryl containing 0-3 heteroatoms, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl or benzyl is unsubstit
  • R c and R d are each independently selected from hydrogen, halogen, carbonyl, hydroxyl, cyano, nitro, phenyl, benzyl, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkoxy, C 3 -C 6 cycloalkyl or halogenated C 3 -C 6 cycloalkyl;
  • n and n are independently selected from 0, 1, 2 or 3.
  • Another object of the present invention is to provide a compound as shown in the general formula II or a pharmaceutically acceptable salt, tautomer, mesomer, racemate, enantiomer, diastereomer thereof:
  • X is selected from -CH2- , -NH-, -O-, -S- or -Se-;
  • Y is selected from carbon or nitrogen
  • Z is selected from -CH2- , -CD2- , -C(O)- or -C(S)-;
  • each R 1 is independently selected from hydrogen, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, halogen, cyano, nitro, -C(O)NR a R b , -C(O)R a , -C(O)OR a , -OR a , -OC(O)R a , -OC(O)OR a , -OC(O)NR a R b , -NR a R b , -SR a , -S(O)R a , -S(O) 2 R a , or a 3-10 membered cycloalkyl, heterocyclyl, aryl and heteroaryl group containing 0-3 heteroatoms, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl,
  • each R2 is independently selected from hydrogen, C1 - C6 alkyl, haloC1- C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, cyano, nitro, -C(O) NRaRb , -C(O) Ra , -C(O ) ORa , -ORa, -OC(O) Ra , -OC (O) ORa , -OC ( O ) NRaRb, -NRaRb , -SRa , -S(O) Ra , -S(O) 2Ra , or 3-10 membered cycloalkyl, heterocyclyl , aryl and heteroaryl containing 0-3 heteroatoms, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are unsubstituted or optionally substituted with
  • each R3 is independently selected from hydrogen, C1 - C6 alkyl, haloC1- C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, cyano, nitro, -C(O) NRaRb , -C(O) Ra , -C(O ) ORa , -ORa, -OC(O) Ra , -OC (O) ORa , -OC ( O ) NRaRb, -NRaRb , -SRa , -S(O) Ra , -S(O) 2Ra , or 3-10 membered cycloalkyl, heterocyclyl , aryl and heteroaryl containing 0-3 heteroatoms, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are unsubstituted or optionally substituted with
  • R4 is selected from hydrogen, deuterium, C1 - C12 alkyl
  • R 5 is selected from hydrogen, deuterium, C 1 -C 12 alkyl
  • R6 , R7 , R8 , R9 , R10 , and R11 are each independently selected from hydrogen, C1 - C6 alkyl, halogenated C1- C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, cyano, nitro, -C(O) NRaRb , -C(O) Ra , -C(O) ORa , -ORa , -OC (O) Ra , -OC (O) ORa , -OC(O) NRaRb , -NRaRb , -SRa , -S (O )Ra, -S(O)2Ra , or 3-10 membered cycloalkyl, heterocyclyl, aryl and heteroaryl containing 0-3 heteroatoms, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocycly
  • Ra and Rb are each independently selected from hydrogen, C1 - C6 alkyl, C2 - C6 alkenyl, C2 - C6 alkynyl, halogen, hydroxyl, cyano, nitro, benzyl, -C(O) NRcRd , -C(O) Rc , -C(O) ORc , -ORc , -OC(O) Rc , -OC(O) ORc , -OC(O) NRcRd , -NRcRd , -SRc , -S(O ) Rc , -S(O) 2Rc , or 3-10 membered cycloalkyl, heterocyclyl , aryl and heteroaryl containing 0-3 heteroatoms, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl or benzyl is unsubstit
  • R c and R d are each independently selected from hydrogen, halogen, carbonyl, hydroxyl, cyano, nitro, phenyl, benzyl, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkoxy, C 3 -C 6 cycloalkyl or halogenated C 3 -C 6 cycloalkyl;
  • n and n are independently selected from 0, 1, 2 or 3.
  • R 1 , X, Y, Z, m, n As defined in Formula I.
  • X is selected from -NH- or -O-.
  • Each R 1 is independently selected from hydrogen, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkoxy, C 2 -C 6 alkenyl, C 2 -C 6 alkynyl, halogen, cyano, hydroxyl, nitro, C 1 -C 10 aryl or -NR a R b ;
  • R a and R b are independently selected from hydrogen, C 1 -C 6 alkyl or -C(O)R c ;
  • R c is selected from hydrogen, halogen, C 1 -C 6 alkyl, halogenated C 1 -C 6 alkyl, C 1 -C 6 alkoxy, halogenated C 1 -C 6 alkoxy or C 3 -C 6 cycloalkyl.
  • Each R 1 is independently selected from hydrogen, methyl, fluorine, chlorine, methoxy, trifluoromethoxy, nitro, cyano, amino, hydroxyl, vinyl, ethynyl, isobutyl, phenyl,
  • the above-mentioned compounds of general formula I of the present invention may also exist in the form of their salts, which are converted in vivo into compounds of general formula I.
  • the compounds of the present invention are converted into pharmaceutically acceptable salt forms according to processes known in the art, and they are used in the form of salts.
  • the pharmaceutically acceptable salts include, but are not limited to, acid addition salts formed between the compound of formula I and the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, naphthalenesulfonic acid, citric acid, tartaric acid, lactic acid, pyruvic acid, acetic acid, maleic acid or succinic acid, fumaric acid, salicylic acid, phenylacetic acid, mandelic acid; and also include acid salts formed between the compound of formula I and inorganic bases.
  • the pharmaceutically acceptable salts include, but are not limited to, basic metal cation salts, alkaline earth metal cation salts, and ammonium cation salts.
  • the compounds of the general formula I of the present invention are preferably the following compounds:
  • Another object of the present invention is to provide a pharmaceutical composition, which comprises a therapeutically effective amount of a compound of Formula I, Formula II or a pharmaceutically acceptable salt, tautomer, mesomer, racemate, enantiomer, diastereomer thereof and a pharmaceutically acceptable carrier or excipient.
  • the pharmaceutical composition of the present invention can be administered in various known ways, such as orally, parenterally, by inhalation spray or via an implanted reservoir.
  • the pharmaceutical composition of the present invention can be administered alone or in combination with other drugs.
  • Oral compositions can be in any orally acceptable dosage form, including but not limited to tablets, capsules, emulsions, suspensions, dispersions and solutions.
  • Commonly used pharmaceutically acceptable carriers or excipients include stabilizers, diluents, surfactants, lubricants, antioxidants, adhesives, colorants, fillers, emulsifiers, etc.
  • Sterile injectable compositions can be formulated using suitable dispersing agents or wetting agents and suspending agents according to techniques known in the art.
  • Pharmaceutically acceptable carriers and solvents that can be used include water, mannitol, sodium chloride solution, and the like.
  • the actual dosage level of the active ingredient in the pharmaceutical composition of the present invention can be varied to obtain an amount of the active ingredient that is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.
  • the dosage level selected depends on a variety of factors, including the activity of the specific compound of the present invention or its salt used, the route of administration, the time of administration, the excretion rate of the specific composition used, the duration of treatment, other drugs, compounds and/or materials used in combination with the specific composition used, the age, sex, weight, general health and previous medical history of the patient being treated, and similar factors well known in the medical field.
  • Another object of the present invention is to provide the use of the compounds of formula I, formula II or their pharmaceutically acceptable salts, tautomers, mesomers, racemates, enantiomers, diastereomers in the preparation of drugs for treating or preventing diseases associated with GSPT1, IKZF1, IKZF2, IKZF3, CK1 ⁇ , N-MYC or C-MYC protein mutations, expression imbalances, allostery and functional abnormalities.
  • the related diseases are cancer, aging, immune diseases, and neurological diseases; wherein the cancer is selected from acute myeloid leukemia, liver cancer, acute lymphocytic leukemia, bladder cancer, bone cancer, brain cancer, breast cancer, cervical cancer, choriocarcinoma, chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), colon cancer, esophageal cancer, gallbladder cancer, gastric cancer, gastrointestinal stromal tumors, head and neck cancer, Hodgkin's lymphoma, laryngeal cancer, leukemia, lung cancer, melanoma, mesothelioma, multiple myeloma, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, kidney cancer, sarcoma, skin cancer, small cell lung cancer, testicular cancer, pharyngeal cancer, thyroid cancer or uterine cancer.
  • the cancer is selected from acute myeloid leukemia, liver cancer, acute lymphocytic le
  • the present invention also provides the use of the compounds of Formula I, Formula II or their pharmaceutically acceptable salts, tautomers, mesomers, racemates, enantiomers, and diastereomers in the preparation of GSPT1 degradation agents.
  • the benzo six-membered heterocyclic compounds of the present invention such as A, B, and C series, have significantly improved degradation effects on GSPT1 protein and anti-proliferation inhibitory activity of AML cells.
  • the degradation percentage of GSPT1 protein was higher than 80%.
  • the in vitro rat liver microsome metabolic half-life of compounds A-1, A-3, and A-10 is about 2-5 times that of the clinical drug CC-90009, and has excellent metabolic stability.
  • the compounds of the present invention can be used to prepare drugs for treating or preventing diseases associated with GSPT1, IKZF1, IKZF2, IKZF3, CK1 ⁇ , N-MYC or C-MYC protein mutations, expression imbalances, conformational changes and functional abnormalities; and can be used to prepare GSPT1 degraders.
  • isomer includes enantiomeric, diastereomeric, and geometric (or conformational) isomeric forms of a given structure.
  • the present application includes R and S configurations for each asymmetric center, Z and E double bond isomers, Z and E conformational isomers, single stereochemical isomers and enantiomers, diastereomers, and geometric (or conformational) isomer mixtures.
  • Suitable acid addition salts are formed from acids, which form non-toxic salts, such as hydrochlorides/chlorides.
  • Suitable base salts are formed from bases, which form non-toxic salts, such as calcium and sodium salts. Hemi-salts of acids and bases, such as hemisulphate and hemicalcium salts, can also be formed.
  • terapéuticaally effective amount refers to an amount of a compound of the present application that (i) treats a specific disease, condition or disorder; (ii) alleviates, relieves or eliminates one or more symptoms of a specific disease, condition or disorder; or (iii) prevents or delays the onset of one or more symptoms of a specific disease, condition or disorder described herein.
  • pharmaceutically acceptable carrier or excipient refers to a non-toxic carrier, adjuvant or vehicle that does not destroy the pharmacological activity of the compound formulated therewith.
  • alkyl refers to a saturated aliphatic hydrocarbon group, and the present invention preferably contains alkyl groups of 1 to 6 carbon atoms.
  • lower alkyl groups containing 1 to 6 carbon atoms include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl,
  • alkenyl refers to an aliphatic hydrocarbon having at least one carbon-carbon double bond, including straight and branched chains having at least one carbon-carbon double bond.
  • the alkenyl groups of the present invention preferably have 2 to 6 carbon atoms.
  • C2-6 alkenyl includes straight or branched unsaturated groups (having at least one carbon-carbon double bond) of 2 to 6 carbon atoms, including but not limited to vinyl, 1-propenyl, 2-propenyl (allyl), isopropenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, etc.
  • alkynyl refers to an aliphatic hydrocarbon having at least one carbon-carbon triple bond, including straight and branched chains having at least one carbon-carbon triple bond.
  • the alkynyl groups of the present invention have 2 to 6 carbon atoms.
  • C2-6 alkynyl includes straight or branched hydrocarbon chain alkynyl groups as defined above having 2 to 6 carbon atoms.
  • alkoxy refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein alkyl is as defined above.
  • alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy.
  • cycloalkyl refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, and the cycloalkyl of the present invention preferably contains 3 to 10 carbon atoms.
  • monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.
  • polycyclic cycloalkyls include spirocyclic, condensed and bridged cycloalkyls.
  • spirocycloalkyl refers to a polycyclic group that shares a carbon atom (called spiral atom) between 5 to 20 monocyclic rings, which may contain one or more double bonds, but no ring has a completely conjugated ⁇ electron system.
  • spiral atom a carbon atom
  • it is 6 to 14 yuan, more preferably 7 to 10 yuan (e.g., 7, 8, 9 or 10 yuan).
  • the spirocycloalkyl is divided into a single spiral cycloalkyl, a double spiral cycloalkyl or a multi-spirocycloalkyl, preferably a single spiral cycloalkyl and a double spiral cycloalkyl, more preferably a 4 yuan/4 yuan, 4 yuan/5 yuan, 4 yuan/6 yuan, 5 yuan/5 yuan or 5 yuan/6 yuan single spiral cycloalkyl.
  • condensed cycloalkyl refers to a 5 to 20-membered, all-carbon polycyclic group in which each ring in the system shares a pair of adjacent carbon atoms with other rings in the system, wherein one or more rings may contain one or more double bonds, but no ring has a completely conjugated ⁇ electron system.
  • it is 6 to 14 members, more preferably 7 to 10 members.
  • it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic condensed cycloalkyl, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic alkyl.
  • bridged cycloalkyl refers to a 5-20 membered, all-carbon polycyclic group in which any two rings share two carbon atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a completely conjugated ⁇ electron system. It is preferably 6-14 members, and more preferably 7-10 members. According to the number of constituent rings, it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged cycloalkyl, preferably bicyclic, tricyclic or tetracyclic, and more preferably bicyclic or tricyclic.
  • heterocyclyl refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which are heteroatoms selected from nitrogen, oxygen or S(O)m (wherein m is an integer from 0 to 2), but excluding the ring part of -O-O-, -O-S- or -S-S-, and the remaining ring atoms are carbon.
  • it contains 3 to 10 ring atoms, of which 1 to 4 are heteroatoms; most preferably, it contains 3 to 8 ring atoms, of which 1 to 3 are heteroatoms; most preferably, it contains 5 to 6 ring atoms, of which 1 to 2 or 1 to 3 are heteroatoms.
  • monocyclic heterocyclic radicals include pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, etc., preferably tetrahydropyranyl, piperidinyl, pyrrolidinyl.
  • Polycyclic heterocyclic radicals include spiro heterocyclic radicals, condensed heterocyclic radicals and bridged heterocyclic radicals.
  • spiro heterocyclic group refers to a polycyclic heterocyclic group in which one atom (called a spiro atom) is shared between 5 to 20 monocyclic rings, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O)m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon. It may contain one or more double bonds, but no ring has a completely conjugated ⁇ electron system. It is preferably 6 to 14 yuan, more preferably 7 to 10 yuan.
  • the spiro heterocyclic group is divided into a single spiro heterocyclic group, a double spiro heterocyclic group or a multi-spiro heterocyclic group, preferably a single spiro heterocyclic group and a double spiro heterocyclic group. More preferably, it is a 4-yuan/4-yuan, 4-yuan/5-yuan, 4-yuan/6-yuan, 5-yuan/5-yuan or 5-yuan/6-yuan single spiro heterocyclic group.
  • fused heterocyclic group refers to a polycyclic heterocyclic group of 5 to 20 members, each ring in the system shares a pair of adjacent atoms with other rings in the system, one or more rings may contain one or more double bonds, but no ring has a completely conjugated ⁇ electron system, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O)m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon.
  • it is 6 to 14 members, more preferably 7 to 10 members.
  • the number of constituent rings it can be divided into a bicyclic, tricyclic, tetracyclic or polycyclic fused heterocyclic group, preferably a bicyclic or tricyclic, more preferably a 5-membered/5-membered or 5-membered/6-membered bicyclic fused heterocyclic group.
  • bridged heterocyclic group refers to a polycyclic heterocyclic group of 5 to 14 members, any two rings sharing two atoms that are not directly connected, which may contain one or more double bonds, but none of the rings has a completely conjugated ⁇ electron system, wherein one or more ring atoms are heteroatoms selected from nitrogen, oxygen or S(O)m (wherein m is an integer from 0 to 2), and the remaining ring atoms are carbon.
  • it is 6 to 14 members, more preferably 7 to 10 members.
  • the number of constituent rings it can be divided into bicyclic, tricyclic, tetracyclic or polycyclic bridged heterocyclic groups, preferably bicyclic, tricyclic or tetracyclic, more preferably bicyclic or tricyclic.
  • the heterocyclic group includes the heterocyclic group as described above (including monocyclic, spiro heterocyclic, fused heterocyclic and bridged heterocyclic) fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, and non-limiting examples thereof include:
  • aryl refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (ie, rings which share adjacent pairs of carbon atoms) group having a conjugated ⁇ electron system, preferably 6- to 10-membered, such as phenyl and naphthyl.
  • heteroaryl refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen.
  • the heteroaryl group is preferably 5 to 10-membered, containing 1 to 3 heteroatoms; more preferably 5 or 6-membered, containing 1 to 2 heteroatoms; preferably, for example, imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, tetrazolyl, pyridyl, pyrimidinyl, thiadiazole, pyrazinyl and pyridazinyl.
  • the heteroaryl group includes a heteroaryl group as described above fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, and non-limiting examples thereof include:
  • hydroxyalkyl refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.
  • haloalkyl refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
  • haloalkoxy refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.
  • Figure 1 shows that the degradation effect of compound A-10 of the present invention on GSPT1 in AML cells is concentration-dependent.
  • Figure 1A shows the degradation effect of compound A-10 and clinical drug CC-90009 on GSPT1 in KG-1 cells;
  • Figure 1B shows the quantitative results of GSPT1 protein in Figure A;
  • Figure 1C shows the degradation effect of compound A-10 and clinical drug CC-90009 on GSPT1 in MV4-11 cells;
  • Figure 1D shows the quantitative results of GSPT1 protein in Figure C.
  • FIG. 2 shows that the degradation effect of compound A-10 of the present invention on GSPT1 in AML cells is time-dependent.
  • FIG3 shows the results of a study on the metabolic stability of compounds A-1, A-3, A-10, A-42, B-1 of the present invention and the clinical drug CC-90009 in rat liver microsomes in vitro.
  • Figure 4 shows the results of pharmacokinetic studies of compounds A-1, A-10 and A-42 of the present invention in rats; wherein, Figures 4A and 4B respectively show the drug-time curve results of CC-90009 after intravenous and oral administration; Figures 4C and 4D respectively show the drug-time curve results of A-1 after intravenous and oral administration; Figures 4E and 4F respectively show the drug-time curve results of A-10 after intravenous and oral administration; Figures 4G and 4H respectively show the drug-time curve results of A-42 after intravenous and oral administration.
  • FIG5 is an in vivo pharmacodynamic study of compound A-10 of the present invention in a MV411 xenograft mouse model;
  • FIG5A is the body weight of mice in each group;
  • FIG5B is the tumor growth curve of mice in each group
  • FIG5C is the tumor mass of mice in each group
  • FIG5D is the tumor tissue dissection of mice in each group.
  • FIG6 is an in vivo pharmacodynamic study of compound A-42 of the present invention in a MV411 xenograft mouse model;
  • FIG6A is the body weight of mice in each group;
  • FIG6B is the tumor growth curve of mice in each group.
  • Figure 7 is an in vivo pharmacodynamic study of compound A-10 of the present invention in a 22RV1 xenograft mouse model; wherein, Figure 7A is the body weight of mice in each group; Figure 7B is the tumor growth curve of mice in each group; Figure 7C is the tumor mass of mice in each group; and Figure 7D is the tumor tissue dissection of mice in each group.
  • Step 1 Dissolve methyl 4-bromo-2-methylbenzoate (10.00 g, 43.86 mmol) in 200 mL of carbon tetrachloride, add N-bromosuccinimide (15.61 g, 87.73 mmol) and azobisisobutyronitrile (0.72 g, 4.38 mmol) in sequence, and heat to reflux and stir for 12 hours. The reaction is complete as monitored by TLC, and the mixture is cooled to room temperature and filtered.
  • Step 2 Dissolve methyl 4-bromo-2-(bromomethyl)benzoate (8.00 g, 26.15 mmol) in 60 mL of N,N-dimethylformamide, add 3-aminopiperidine-2,6-dione hydrochloride (5.15 g, 31.38 mmol) and DIPEA (8.45 g, 65.38 mmol) in sequence, and heat to 90°C under nitrogen protection and stir for 4 hours.
  • reaction solution is poured into 200 mL of water to precipitate a light blue solid, which is allowed to stand for complete precipitation, and filtered, and the filter cake is vacuum dried to obtain a light blue solid compound 3: 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (5.89 g, yield 70.2%).
  • Step 3 Dissolve 3-(5-bromo-1-oxoisoindolin-2-yl)piperidine-2,6-dione (5.00 g, 15.53 mmol) in 50 mL of dry N,N-dimethylformamide, and add 1,1'-bis(diphenylphosphino)ferrocene (0.40 g, 0.71 mmol), tris(dibenzylideneacetone)dipalladium (0.39 g, 0.43 mmol) and zinc cyanide (1.88 g, 16.00 mmol) in sequence. After the addition is complete, heat to 90°C and stir for 4 hours under nitrogen protection.
  • Step 4 2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindoline-5-carbonitrile (3.50 g, 13.00 mmol) was dissolved in 40 mL of N,N-dimethylformamide, and di-tert-butyl dicarbonate (5.67 g, 26.00 mmol) and 15 mL of Raney nickel were added in sequence. After the addition was completed, the mixture was heated to 50°C and stirred for 5 hours under full replacement of hydrogen. The reaction was complete as monitored by TLC, and the filter cake was washed with ethyl acetate.
  • Step 5 Dissolve tert-butyl ((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)carbamate (3.00 g, 8.04 mmol) in 30 mL of hydrochloric acid-dioxane solution (4 M) and stir at room temperature for 3 hours. The reaction was complete as monitored by TLC, and the filter cake was filtered and vacuum dried to obtain a light yellow solid, which was Intermediate-1: 3-(5-aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (1.80 g, yield 72.4%), which was used directly in the next step without further purification.
  • Step 1 Dissolve 2-hydroxybenzaldehyde (2.00 g, 16.39 mmol) in 15 mL of acrylonitrile, add 1,4-diazidebicyclo[2.2.2]octane (1.84 g, 16.39 mmol), and heat to reflux under nitrogen protection for 3 hours.
  • Step 2 Dissolve 2H-chromene-3-carbonitrile (2.30 g, 14.73 mmol) in 40 mL of 10% sodium hydroxide aqueous solution, heat to 60°C and stir for 10 hours. After TLC monitoring, the reaction was complete, and dilute hydrochloric acid (2 M) was added to adjust the pH to 4-5, and a white solid was precipitated. The solid was allowed to stand for complete precipitation, and then filtered and dried to obtain a yellow solid compound 8: 2H-chromene-3-carboxylic acid (1.65 g, yield 63.7%).
  • Step 3 Dissolve 2H-chromene-3-carboxylic acid (1.50 g, 8.52 mmol) in 20 mL of methanol, add 10% Pd/C (1.00 g), stir at room temperature for 8 hours. The reaction was complete as monitored by TLC, filtered, the filter cake was washed with methanol, and the filtrate was concentrated in vacuo to obtain a brown solid, which is Intermediate-2: chroman-3-carboxylic acid (0.76 g, yield 50.2%).
  • Step 1 Dissolve 4-bromo-3-fluorobenzoic acid (9, 15.00 g, 68.49 mmol) in 150 mL of anhydrous tetrahydrofuran under nitrogen protection. Cool to -70 ° C, add lithium diisopropylamide (72 mL, 143.8 mmol) dropwise, continue to react for 1 hour after the addition is complete, add iodomethane (29.16 g, 205.5 mmol) dropwise, move to room temperature and react for 12 hours. The reaction is complete as monitored by TLC, and 150 mL of saturated ammonium chloride solution is added dropwise to the reaction solution.
  • Step 2 Dissolve 4-bromo-3-fluoro-2-methylbenzoic acid (10, 9.67 g, 41.50 mmol) in 80 mL of N, N-dimethylformamide, add potassium carbonate (11.47 g, 83.01 mmol) and iodomethane (8.84 g, 62.26 mmol) in turn, and react at room temperature for 2 hours.
  • Step 3 Dissolve methyl 4-bromo-3-fluoro-2-methylbenzoate (11, 9.29 g, 37.60 mmol) in 150 mL of chloroform, add N-bromosuccinimide (16.73 g, 94.00 mmol) and azobisisobutyronitrile (1.85 g, 11.28 mmol) in sequence, and heat to reflux for 10 hours. The reaction was complete as monitored by TLC, and the filtrate was vacuum-filtered and concentrated. Petroleum ether was added for slurrying to obtain a white solid 12: methyl 4-bromo-2-(bromomethyl)-3-fluorobenzoate (10.36 g, yield 84.5%).
  • Step 4-Step 7 Using the raw material 4-bromo-2-(bromomethyl)-3-fluorobenzoic acid methyl ester, refer to the synthesis method of intermediate-1 to obtain white solid intermediate-3: 3-(5-(aminomethyl)-4-fluoro-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride.
  • reaction solution was poured into 30 mL of ice water to precipitate a brown solid.
  • the solid was allowed to stand for complete precipitation, and then filtered.
  • reaction solution was poured into 30 mL of ice water to precipitate a brown solid.
  • the solid was allowed to stand for complete precipitation, and then filtered.
  • Synthesis of compound A-3 Using raw material 6-fluoro-2H-chromene-3-carboxylic acid, refer to the synthesis method of compound A-1 in Example 1 to obtain a white solid, namely A-3 (0.12 g, yield 83.9%, purity 98.72%).
  • Step 1 3-Formyl-4-hydroxybenzonitrile (25, 1.00 g, 6.80 mmol) was dissolved in 20 mL of 1,4-dioxane, potassium carbonate (0.94 g, 6.80 mmol) and acrolein (0.76 g, 13.60 mmol) were added in sequence, and the mixture was heated to reflux and stirred for 5 hours. The reaction was completed as monitored by TLC, and the reaction solution was poured into 20 mL of water, extracted with ethyl acetate, and the organic phases were combined and washed with 2M sodium hydroxide solution and saturated brine in sequence, dried over anhydrous sodium sulfate, and concentrated in vacuo.
  • Step 2 3-Formyl-2H-chromene-6-carbonitrile (1.03 g, 5.57 mmol) was dissolved in 20 mL of tetrahydrofuran, and sodium chlorite (5.03 g, 55.66 mmol) and sodium dihydrogen phosphate (6.68 g, 55.66 mmol) dissolved in 15 mL of water were added dropwise, and 2-methyl-2-butene (4.27 g, 60.81 mmol) was added. After the addition, the mixture was stirred at room temperature for 4 hours. The reaction was monitored by TLC to be complete. The reaction solution was concentrated in vacuo and diluted with 30 mL of water.
  • Step 3 Prepare the raw material 6-cyano-2H-chromene-3-carboxylic acid according to the synthetic method of compound A-1 to obtain white solid A-11: (0.08 g, yield 54.8%, purity 96.95%).
  • Step 1 N-acetyl-p-aminophenol (28, 10.00 g, 66.20 mmol) was dissolved in 40 mL of trifluoroacetic acid, and hexamethylenetetramine (36.56 g, 0.26 mol) was added to the reaction solution in batches under an ice bath. After the addition was completed, the temperature was raised to 70°C and stirred for 5 hours. The reaction was monitored by TLC to be complete. After the reaction solution was cooled to room temperature, it was poured into 200 mL of water, extracted with ethyl acetate, the organic phases were combined and washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated in vacuo.
  • Step 2 Dissolve N-(3-formyl-4-hydroxyphenyl)acetamide (2.03 g, 11.34 mmol) in ethanol (30 mL) and dilute hydrochloric acid (6 M, 10 mL), heat to reflux and stir for 8 hours. The reaction was complete as monitored by TLC, the reaction solution was concentrated in vacuo, and slurried with ethanol to obtain a black solid 30: 5-amino-2-hydroxybenzaldehyde hydrochloride (1.93 g, yield 98.5%).
  • Step 3 Dissolve 5-amino-2-hydroxybenzaldehyde hydrochloride (1.93 g, 11.15 mmol) in 30 mL of tetrahydrofuran, add triethylamine (1.13 g, 11.15 mmol) and di-tert-butyl dicarbonate (2.92 g, 13.39 mmol) in turn, and stir at room temperature for 16 hours. The reaction was complete as monitored by TLC, and the reaction solution was concentrated in vacuo.
  • Step 4-Step 6 Using the raw material (3-formyl-4-hydroxyphenyl)carbamic acid tert-butyl ester, refer to the synthesis method of intermediate-2 and compound A-1 to obtain white solid 34: ((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)carbamoyl)-2H-chromen-6-yl)carbamic acid tert-butyl ester (1.21 g, yield 20.1%).
  • Step 7 Dissolve tert-butyl ((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)carbamoyl)-2H-chromen-6-yl)carbamate (1.21 g, 2.22 mmol) in 30 mL of hydrochloric acid-dioxane solution (4 M) and stir at room temperature for 2 hours. The reaction was complete as monitored by TLC.
  • reaction solution was concentrated in vacuo and the residue was adjusted to weak alkalinity with saturated sodium bicarbonate solution, filtered, and the filter cake was washed with a small amount of petroleum ether and dried in vacuo to obtain a white solid A-12 (0.99 g, yield 95.0%, purity 98.70%).
  • Step 1 5-bromo-2-hydroxybenzaldehyde (36, 1.50 g, 7.50 mmol) was dissolved in 20 mL of N, N-dimethylformamide, and lithium chloride (0.95 g, 22.51 mmol), tetrakis(triphenylphosphine)palladium (0.87 g, 0.75 mmol) and tributyl vinyl tin (2.85 g, 9.00 mmol) were added in sequence. After the addition was completed, the temperature was raised to 95°C and stirred for 4 hours. The reaction was monitored by TLC to be complete, and the reaction solution was poured into 100 mL of ice water to precipitate a brown solid.
  • Step 2-Step 4 Using the raw material 2-hydroxy-5-vinylbenzaldehyde, refer to the synthesis method of intermediate-2 and compound A-1 to obtain white solid A-14 (0.07 g, yield 50.5%, purity 98.49%).
  • Step 1 Dissolve 5-bromo-2-hydroxybenzaldehyde (36, 1.80 g, 9.00 mmol) in 30 mL of water, add phenylboric acid (1.65 g, 13.50 mmol), potassium carbonate (2.49 g, 18.00 mmol) and tetrakis(triphenylphosphine)palladium (0.02 g, 0.02 mmol) in sequence, and add until complete. After nitrogen protection, the reaction solution was heated to 80 ° C and stirred for 6 hours. The reaction was monitored by TLC.
  • Step 2-Step 4 Use the raw material 4-hydroxy-[1,1'-biphenyl]-3-carboxaldehyde to refer to the synthesis method of intermediate-2 and compound A-1 to obtain white solid A-17 (0.10 g, yield 60.8%, purity 98.56%).
  • N-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-5-yl)methyl)-6-amino-2H-chromene-3-carboxamide (Compound 12, 0.07 g, 0.16 mmol) was dissolved in 5 mL of N,N-dimethylformamide, and glycolic acid (0.01 g, 0.19 mmol) and HATU (0.09 g, 0.24 mmol) were added in sequence. After the addition, the mixture was stirred at room temperature for 1 hour. DIPEA (0.06 g, 0.47 mmol) was added and the mixture was stirred at room temperature overnight.
  • Step 1 3-Chloro-4-methyl-phenol (51, 1.00 g, 7.04 mmol) was dissolved in 20 mL of acetonitrile, and formaldehyde (1.11 g, 36.99 mmol), triethylamine (3.74 g, 36.99 mmol) and magnesium chloride (2.68 g, 28.16 mmol) were added in sequence. After addition, the mixture was heated to 80°C and stirred for 15 hours. The reaction was monitored by TLC to be complete, and 50 mL of hydrochloric acid (1 M) was added to quench the mixture, and the mixture was extracted with ethyl acetate.
  • Step 2-Step 4 Use the raw material 4-chloro-2-hydroxy-5-methylbenzaldehyde to refer to the synthesis method of intermediate-2 and compound A-1 to obtain white solid A-29 (0.03 g, yield 19.4%, purity 98.23%).
  • Step 1-Step 6 Use the raw material methyl 2-bromo-6-methylbenzoate (64) and refer to the synthesis method of intermediate-1 and compound A-1 to obtain white solid A-39 (0.10 g, yield 57.1%, purity 98.16%).
  • Step 1-Step 6 Use the raw material 5-bromo-2-methylbenzoic acid methyl ester (70) to prepare according to the synthesis method of intermediate-1 and compound A-1 to obtain white solid A-40 (0.10 g, yield 62.7%, purity 99.20%).
  • Step 1-Step 6 Using raw material methyl 3-bromo-2-methylbenzoate (76) and referring to the synthesis method of intermediate-1 and compound A-1, a white solid A-41 (0.09 g, yield 54.2%, purity 97.59%) was obtained.
  • the raw material 1,2,3,4-tetrahydroquinoline-3-carboxylic acid was used to prepare the product according to the synthetic method of compound A-1 to obtain a white solid C-1 (0.09 g, yield 63.3%, purity 98.67%).
  • KG-1 cells (ATCC) were grown in IMDM (containing 20% FBS) medium. After centrifugation and counting, the cell concentration was adjusted to 10 6 cells/well and plated in a 6-well plate, with 1350 ⁇ l per well. 150 ⁇ l of DMSO and the test compound of the present invention were added, the compound concentration was 200 nM, and the cells were placed in a 5% CO 2 , 37°C incubator for 4 hours. The cells were centrifuged to discard the culture medium, and PBS was added for washing and discarded.
  • IMDM containing 20% FBS
  • Whole cell lysates were prepared with a mixture of protease inhibitors (100 ⁇ , Fude Biotechnology), a mixture of protein phosphatase inhibitors (100 ⁇ , Fude Biotechnology), super nuclease (Biyuntian) and high-strength RIPA lysis buffer (Thermo Fisher), and placed on ice for 30 minutes. The cell debris precipitate was discarded by centrifugation, and the supernatant whole cell lysate was collected and transferred to a new centrifuge tube. After BCA protein determination, the sample was prepared using 5X loading buffer (Thermo Fisher).
  • the samples were electrophoresed in 4-20% precast gel (SDS-PAGE gel) to separate proteins, then transferred to PVDF membrane and blocked with 5% NFDM/TBST at room temperature for 1 hour, incubated with primary antibody at 4°C overnight, and incubated with secondary antibody at room temperature for 2 hours the next day.
  • the signal was detected using MINICHEMTM imaging system.
  • Anti-GSPT1 Abcam ab234433
  • the degradation effects of the compounds on GSPT1 protein are shown in Table 1, wherein A indicates that the degradation percentage of GSPT1 protein is not less than 80%, B indicates that the degradation percentage is less than 80% but not less than 50%, C indicates that the degradation percentage is less than 50% but not less than 25%, and D indicates that the degradation percentage is less than 25%.
  • the benzo six-membered heterocyclic compounds of the present invention such as A, B, and C series, have significantly improved degradation effects on GSPT1 protein, while the benzo six-membered aliphatic ring loses the degradation effect on GSPT1 protein.
  • the degradation effect of the compound A-10 of the present invention on GSPT1 in KG-1 and MV4-11 cells is concentration-dependent and time-dependent, and the effect is more significant than that of CC-90009.
  • KG-1 cells in the logarithmic growth phase were diluted with culture medium (RPMI + 10% FBS), plated on 96-well plates at 5000 cells/well, and cultured in a 5% CO 2 , 37°C incubator for 24 hours.
  • the test compound of the present invention was prepared into a 10mM stock solution with DMSO, diluted with culture medium to set 9 concentration gradients, 3 replicates for each concentration, and cultured in a 5% CO 2 , 37°C incubator for 72 hours after drug addition. Cell proliferation was detected using CCK8 method.
  • IC 50 value of the compound's inhibition of cell growth was determined by enzyme labeling. IC 50 value was calculated using GraphPad Prism 7 software.
  • the anti-KG-1 cell proliferation activities of the compounds are shown in Table 2, wherein A represents IC 50 ⁇ 1 nM, B represents 1 ⁇ IC 50 ⁇ 10 nM, C represents 10 nM ⁇ IC 50 ⁇ 100 nM, D represents 100 nM ⁇ IC 50 ⁇ 1000 nM, and E represents IC 50 ⁇ 1000 nM.
  • the anti-proliferation inhibitory activity of the benzo six-membered heterocyclic compounds of the present invention is significantly improved, while the benzo six-membered aliphatic ring loses its anti-proliferation inhibitory effect on KG-1 cells.
  • Biological test example 3 The compounds of the present invention have antiproliferative activity against various tumor cells
  • U937, MOLM-13, and MV4-11 cells (ATCC) in the logarithmic growth phase were diluted with culture medium, plated on 96-well plates at 5000 cells/well, and cultured in a 5% CO 2 , 37°C incubator for 24 hours.
  • the compound was prepared into a 10mM stock solution with DMSO, diluted with culture medium to set 9 concentration gradients, 3 replicates for each concentration, and cultured in a 5% CO 2 , 37°C incubator for 72 hours after drug addition. Cell proliferation was detected using the CCK8 method.
  • the IC 50 value of the compound's inhibition of cell growth was determined using the enzyme labeling method. The IC 50 value was calculated using GraphPad Prism 7 software.
  • A represents IC 50 ⁇ 1 nM
  • B represents 1 ⁇ IC 50 ⁇ 10 nM
  • C represents 10 nM ⁇ IC 50 ⁇ 100 nM
  • D represents 100 nM ⁇ IC 50 ⁇ 1000 nM
  • E represents IC 50 ⁇ 1000 nM.
  • the benzo six-membered heterocyclic compounds of the present invention such as series A, B, and C, have significantly improved anti-proliferation inhibitory activity against a variety of AML cells, while the benzo six-membered aliphatic ring loses its anti-proliferation inhibitory effect on AML cells.
  • the compound to be tested was prepared into a 10mM stock solution with DMSO, and diluted with acetonitrile and PBS in turn, vortexed, and the DMSO content was controlled at 0.1%; 10 ⁇ L of the diluted working solution was incubated with 65 ⁇ L of liver microsome system (30 ⁇ L 100mM PBS, 25 ⁇ L 20mM MgCl 2 , 10 ⁇ L 5mg/mL RLM), preheated at 37°C for 5min; 25 ⁇ L 4mM NADPH was added to start the reaction; samples were taken at different time points (0min, 5min, 15min, 30min, 60min, 120min), acetonitrile solution containing internal standard was added to terminate the reaction, and after vortexing and centrifugation, the supernatant was collected for LC-MS/MS analysis and the half-life and clearance rate were calculated.
  • RLM was purchased from Red Liver.
  • FIG3 shows the results of a study on the metabolic stability of compounds A-1, A-3, A-10, and A-42 of the present invention in rat liver microsomes in vitro.
  • the half-life of compounds A-1, A-3, A-10, and A-42 is more than 2 times that of the clinical drug CC-90009, and has excellent metabolic stability in rat liver microsomes in vitro.
  • Sample collection and preparation SPF SD rats (male, 210-230g, 42-48 days, red liver) in good growth state were given intravenous 2mg/kg and oral 10mg/kg. Blood samples were collected from the eye sockets at different blood collection times and transferred to centrifuge tubes containing anticoagulants. The venous blood collection time was 0.08, 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours, and the oral blood collection time was 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours. The blood samples were immediately centrifuged at 4°C, 8000rpm, and 10 minutes to collect plasma, which was stored in an ultra-low temperature refrigerator at -80°C until LC-MS/MS analysis.
  • Pharmacokinetic data analysis Pharmacokinetic software was used to calculate the pharmacokinetic parameters of the compound in a non-compartmental model.
  • the peak blood concentration (C max ) and the time to peak blood concentration (T max ) were taken as observed values.
  • the half-life T 1/2 , the area under the drug-time curve AUC last and AUC inf , the apparent distribution volume V ss , the clearance rate CL and other parameters were determined by regression analysis of the linear end part of the logarithmic blood drug concentration-time curve and the linear trapezoidal rule.
  • FIG4 shows the results of pharmacokinetic studies of compounds A-1, A-10, A-42 and clinical drug CC-90009 in rats.
  • the clearance rate of compound A-10 after intravenous administration is one-third of that of CC-90009, and its half-life is longer than that of CC-90009; the initial blood drug concentration is much higher than that of CC-90009, and the plasma drug exposure is nearly four times that of CC-90009; and the exposure after oral administration is nearly three times that of CC-90009, with a higher peak height and a smaller peak time, indicating that compound A-10 is absorbed faster.
  • Compound A-42 has a lower clearance rate and higher intravenous and oral plasma drug exposure, which are fourteen times and four times that of CC-90009, respectively.
  • Biological Test Example 6 In vivo pharmacodynamic study of the compounds of the present invention in an acute myeloid leukemia xenograft mouse model
  • the human leukemia cells MV411 (cell bank of the Chinese Academy of Sciences) used in this experiment were cultured in an IMDM medium supplemented with 10% FBS in a 37°C incubator containing CO 2. Before the cells were cultured for ten consecutive generations, MV411 cells in the logarithmic growth phase were used to adjust the cell concentration to 1 ⁇ 10 8 cells/mL using serum-free culture medium. Under sterile conditions, they were inoculated subcutaneously into BABL/c Nude mice (Jiangsu Jicui Pharmaceutical Kang Biotechnology Co., Ltd., female, 4-6 weeks old, 14-16g) on the right side of the back (near the armpit) with an inoculation volume of 100 ⁇ L/mouse.
  • mice When the average tumor volume of tumor-bearing mice reached ⁇ 100 mm 3 , they were randomly divided into groups, and the tumor volumes of each group were roughly the same.
  • the day of group administration was defined as day 0.
  • the compound was administered to animals in each treatment group by oral gavage at 15 mg/kg, 30 mg/kg, and 60 mg/kg (once a day). The body weight and tumor volume of the animals were measured every other day.
  • Tumor weight TGI (%) ⁇ 1-[(final average tumor weight of the experimental group)/(final average tumor weight of the control group of tumor-bearing mice not treated with the drug)] ⁇ 100%
  • tumor volume TGI (%) [1-relative tumor volume of the experimental group/relative tumor volume of the control group of tumor-bearing mice not treated with the drug]*100%.
  • Figure 5 shows the in vivo pharmacodynamic study of compound A-10 in the MV411 xenograft mouse model. In the absence of significant weight loss in mice, the tumor weight TGI of compound A-10 reached 20.82%, 56.61%, and 59.76% at doses of 15 mg/kg, 30 mg/kg, and 60 mg/kg, respectively, which was significantly better than the clinical molecule CC-90009.
  • Figure 6 shows the in vivo pharmacodynamic study of compound A-42 in the MV411 xenograft mouse model. In the absence of significant weight loss in mice, the tumor volume TGI of compound A-42 reached 100% at a dose of 60 mg/kg, which is comparable to the level of the clinical molecule MRT-2359.
  • the 22RV1 cells (Cell Bank of the Chinese Academy of Sciences) used in this experiment were cultured in 1640 medium supplemented with 10% FBS in a 37°C incubator containing CO2 . Before the cells were cultured for ten consecutive generations, the 22RV1 cells in the logarithmic growth phase were used to adjust the cell concentration to 5 ⁇ 107 /mL using serum-free culture medium. Under sterile conditions, they were inoculated subcutaneously on the right side of the back (near the armpit) of BABL/c Nude mice with an inoculation volume of 100 ⁇ L/mouse.
  • mice When the average tumor volume of tumor-bearing mice reached ⁇ 50-100mm3, they were randomly divided into 4 groups with 8 mice in each group, and the tumor volume of each group was roughly the same.
  • the day of group administration was defined as day 0.
  • the compound was administered to animals in each treatment group by oral gavage at 60 mg/kg (once a day).
  • the body weight and tumor volume of the animals were measured every other day.
  • Tumor weight TGI (%) ⁇ 1-[(final average tumor weight of the experimental group)/(final average tumor weight of the control group of tumor-bearing mice without medication)] ⁇ 100%.
  • Figure 7 shows the in vivo pharmacodynamic study of compound A-10 in the 22RV1 xenograft mouse model.
  • compound A-10 was administered at a dose of 60 mg/kg, the TGI reached 95.10%, which was significantly better than the clinical molecule CC-90009 and the marketed drug Enzalutamide.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Oncology (AREA)
  • Hematology (AREA)
  • Plural Heterocyclic Compounds (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

本发明公开了一种苯并六元杂环类化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体。本发明的化合物对GSPT1蛋白的降解效果、AML细胞的抗增殖抑制活性显著提高。因此，本发明化合物可以用于制备治疗或预防由GSPT1、IKZF1、IKZF2、IKZF3、CK1α、N-MYC或C-MYC蛋白突变、表达失衡、变构与功能异常相关疾病的药物；可以用于制备GSPT1降解剂。

Description

一种苯并六元杂环类GSPT1蛋白降解剂及其应用 技术领域
本发明属于医药技术领域，具体涉及一种苯并六元杂环类GSPT1蛋白降解剂及其制备方法与应用。
 背景技术
靶向蛋白质降解技术(Target Protein Degradation technology，TPD)是通过利用真核细胞中固有的调控蛋白质稳态的蛋白质降解机制，干扰蛋白质功能的新技术。该技术的兴起一定程度上解决了小分子抑制剂和基因干扰技术所面临的困境。TPD目前主要通过泛素蛋白酶体和溶酶体降解目标蛋白，目前该领域发展最为成熟、研究最多的是基于泛素化-蛋白酶体系统的蛋白水解靶向嵌合体(proteolysis-targeting chimera,PROTAC)和分子胶(Molecular glue，MGs)。靶向蛋白质降解技术在诸如恶性肿瘤、神经退行性疾病、代谢性疾病等多种疾病领域具有广阔的应用前景和发展空间。
分子胶是一种诱导接近的小分子，可以对各种生物过程进行精确的时间控制，例如信号转导、转录、染色质调节以及蛋白质折叠、定位和降解。作为接近的化学诱导剂，能诱导E3泛素连接酶与靶蛋白发生蛋白-蛋白相互作用，导致靶蛋白降解。分子胶可以通过形成三元复合物促进两种蛋白质的二聚化或共定位，从而产生多种生物学和药理学功能。
G1到S期转换1蛋白(G1 to S phase transition 1，GSPT1，也被称为eRF3a)是机体内的一种翻译终止因子，GSPT1的下调可导致关键蛋白的异常表达，并在各种肿瘤细胞中抑制增殖或诱导凋亡。自从发现GSPT1是E3泛素连接酶CRBN的新底物后，它可以被分子胶降解剂定向降解。目前已有多种分子胶降解剂正处于临床及临床前研究，这些分子在治疗血液肿瘤以及某些实体肿瘤方面非常有效。GSPT1具有多功能性，除了作为第二类肽链释放因子，参与到蛋白质翻译终止外，它还与细胞凋亡、细胞周期调控、肿瘤的发生发展等过程都相关。例如在细胞凋亡过程中GSPT1会以蛋白质水解的方式加工形成另外一种异构型，该异构型中含有一个保守的N末端凋亡抑制蛋白结合体。GSPT1通过这种加工形式与凋亡抑制因子(inhibitor of apoptosis proteins，IAPs)相互作用，解除对IAP的抑制释放caspases，从而促进细胞凋亡。而eRF3的M结构域则与Polya-binding protein(PABP)蛋白相互作用，偶联了翻译终止和mRNA降解。
急性髓系白血病(acute myeloid leukemia，AML)是一种遗传和生物学上的异质性髓系恶性肿瘤，以未成熟的髓系祖细胞异常扩增为特征，损害机体正常造血功能，从而引起严重的感染、贫血、出血，其特点是生存率低和复发率高。因此作为一种相对罕见的恶性肿瘤，已被美国食品和药物管理局视为孤儿病。GSPT1作为CRL4-CRBN-E3泛素连接酶的新底物，在治疗AML方面发挥着重要作用。在靶向GSPT1用于治疗AML的临床药物如CC-90009、BTX-1188等正在研发。CC-90009主要通过静脉注射的方式用于治疗AML，而BTX-1188是一种可以口服的药物，用于治疗AML和NHL。通过进一步研究发现GSPT1的降解与ATF3和ATF4基因表达上调有关，而ATF3和ATF4基因对综合应激反应通路是非常重要的。综合应激反应通路的激活与eRF2的磷酸化有很大的关系。当综合应激反应通路被激活后会导致细胞的急性凋亡。
据世界卫生组织报道，乳腺癌是女性恶性肿瘤死亡的首要原因。在2018年Wang等分析了他们收集的乳腺癌样品中提取的总RNA中的基因表达，鉴定出了包括GSPT1在内的可作为三阴性乳腺癌潜在治疗靶标的五种基因。最近，Malta-Vacas等的研究也表示较长的GSPT1等位基因12-GGC存在于5.1％的乳腺癌患者中，另外通过mRNA定量实验也表明，相对于正常邻近组织，GSPT1在具有12-GGC等位基因的患者的肿瘤组织中过表达。此外，Miri等进行了一项乳腺癌患者GSPT1基因与乳腺癌易感性的试验，研究发现GSPT1在乳腺癌组织中表达量明显升高，其中GSPT1外显子中较长的等位基因12-GGC的存在可增加乳腺癌的发生风险3倍。因此有学者认为12-GGC等位基因与癌症发展是GSPT1通过调节mRNA降解或翻译效率等使蛋白质发生功能丧失来实现的。
在发展中国家，胃癌是癌症相关死亡的主要原因。Malta-Vacas等发现GSPT1在肠道型胃肿瘤中的表达水平明显高于弥漫型胃肿瘤。除此之外，他们还评估GSPT1与胃癌的潜在遗传易感性的关系发现，无论基因型如何，具有12-GGC等位基因的患者患胃癌的风险较平常人增加了20倍。Tian等探讨了GSPT1对胃癌发生的潜在影响，发现在胃癌组织中GSPT1的表达水平显著增加。有学者认为GSPT1的过表达通过提高特定致癌基因mRNA的翻译效率来促进肿瘤发展，也有学者认为是通过GSPT1的其他作用如调节细胞周期，细胞凋亡等。由于GSPT1还会参与到细胞骨架形成及控制染色体的分离，因此GSPT1对染色体分离和胞质分裂的影响也可能是该基因影响胃癌发生的一种潜在机制。
肝癌是一种常见肿瘤疾病。从临床数据来看，肝癌组织中GSPT1的表达量较正常细胞中要普遍增高。由于GSPT1的主要功能之一是调节细胞由G1期到S期的转变，据此有研究者推测在干扰GSPT1的mRNA表达后可改变细胞周期，从而使G1期细胞百分比增多，导致肿瘤细胞的活力以及增殖能力下降。虽然GSPT1在肝癌的发生发展过程中具有促癌效应，但是也有研究者在通过构建过表达和敲低GSTP1基因的HepG2细胞中发现GSPT1在肝癌的发展过程中可能会抑制癌变进程。
结直肠癌作为消化系统最主要的恶性肿瘤之一。Xiao等的研究证明与正常对照组的结直肠细胞相比，GSPT18在人结直肠癌HCT116细胞中过表达，进一步实验也表明在具有高GSPT1表达的结直肠癌细胞中，敲低GSPT1可以抑制结直肠癌细胞的增殖和迁移，有研究结果显示GSPT1通过mTOR途径抑制HCT116细胞的周期进展，当mTOR途径被激活后，就会导致mRNA亚组的翻译增加。因此GSPT1在结直肠癌的发生发展过程中可能扮演着原癌基因的角色。
综上所述，GSPT1在相关实体肿瘤的发生发展过程中具有重要作用。但由于其功能的多样性，调节机制复杂性，在不同的肿瘤中发挥着不同的作用。但是目前普遍认可的是GSPT1是大多数肿瘤的原癌基因，在某些特定种类的肿瘤中，扮演着抑癌基因的角色。就目前而言GSPT1在肿瘤中的相关作用机制尚缺乏较为深入的研究，但毋庸置疑的是，GSPT1已成为肿瘤治疗中的一个强有力的潜在靶点。
发明内容
发明目的：本发明目的之一在于提供一种如通式I的苯并六元杂环类化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体：
其中：
表示单键或双键；
X选自-CH2-、-NH-、-O-、-S-或-Se-；
Y选自碳或氮；
Z选自-CH2-，-CD2-，-C(O)-或-C(S)-；
每个R1分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
每个R2分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
每个R3分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
R4选自氢、氘、C1-C12烷基、
R5、R6、R7、R8、R9、R10分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
Ra、Rb分别独立地选自氢、C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、羟基、氰基、硝基、苄基、-C(O)NRcRd、-C(O)Rc、-C(O)ORc、-ORc、-OC(O)Rc、-OC(O)ORc、-OC(O)NRcRd、-NRcRd、-SRc、-S(O)Rc、-S(O)2Rc或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基、杂芳基或苄基未被取代或任选地被1-3个Rc取代；
Rc、Rd分别独立地选自氢、卤素、羰基、羟基、氰基、硝基、苯基、苄基、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基、C3-C6环烷基或卤代C3-C6环烷基；
m、n分别独立地选自0、1、2或3。
本发明另一目的在于提供一种如通式II所示的化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体：
其中：
表示单键或双键；
X选自-CH2-、-NH-、-O-、-S-或-Se-；
Y选自碳或氮；
Z选自-CH2-，-CD2-，-C(O)-或-C(S)-；
每个R1分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
每个R2分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
每个R3分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
R4选自氢、氘、C1-C12烷基、
R5选自氢、氘、C1-C12烷基；
R6、R7、R8、R9、R10、R11分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
Ra、Rb分别独立地选自氢、C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、羟基、氰基、硝基、苄基、-C(O)NRcRd、-C(O)Rc、-C(O)ORc、-ORc、-OC(O)Rc、-OC(O)ORc、-OC(O)NRcRd、-NRcRd、-SRc、-S(O)Rc、-S(O)2Rc或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基、杂芳基或苄基未被取代或任选地被1-3个Rc取代；
Rc、Rd分别独立地选自氢、卤素、羰基、羟基、氰基、硝基、苯基、苄基、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基、C3-C6环烷基或卤代C3-C6环烷基；
m、n分别独立地选自0、1、2或3。
本发明的通式I化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体，优选具有式Ia、式Ib或式Ic结构的化合物：
其中，R1、X、Y、Z、m、n、如通式I中的定义。
在某些优选的实施方式中，
X选自-NH-或-O-。
在某些优选的实施方式中，
每个R1分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基、C2-C6烯基、C2-C6炔基、卤素、氰基、羟基、硝基、C1-C10芳基或-NRaRb；
Ra、Rb分别独立地选自氢、C1-C6烷基或-C(O)Rc；
Rc选自氢、卤素、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基或C3-C6环烷基。
在某些更优选的实施方式中，
每个R1分别独立地选自氢、甲基、氟、氯、甲氧基、三氟甲氧基、硝基、氰基、氨基、羟基、乙烯基、乙炔基、异丁基、苯基、
本发明涉及的上述通式I化合物还可以以其盐形式存在，它们在体内转化为通式I化合物。例如，在本发明的范围内，按照本领域已知的工艺，将本发明化合物转化为药学上可接受的盐的形式，并且以盐形式使用它们。
在一些优选的实施方式中，所述药学上可接受的盐包括但不限于通式I化合物与下列酸形成的酸加成盐：盐酸、氢溴酸、硫酸、磷酸、甲磺酸、苯磺酸、对甲苯磺酸、萘磺酸、柠檬酸、酒石酸、乳酸、丙酮酸、乙酸、马来酸或琥珀酸、富马酸、水杨酸、苯基乙酸、杏仁酸；还包括通式I化合物与无机碱形成的酸式盐。
在一些更优选的实施方式中，所述药学上可接受的盐包括但不限于碱性金属阳离子盐、碱土金属阳离子盐和铵阳离子盐。
本发明通式I的化合物优选以下化合物：
本发明另一目的在于提供一种药物组合物，所述药物组合物包括治疗有效量的通式I、通式II的化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体以及药学上可接受的载体或赋形剂。
本发明的药物组合物可以采用各种已知的方式施用，例如口服、胃肠外施用、通过吸入喷雾施用或经由植入的贮库施用。本发明的药物组合物可单独给药也可与其他药物联合用药。口服组合物可以是任何口服可接受的剂型，包含但不限于片剂、胶囊剂、乳剂以及混悬剂、分散物和溶液。常用的药学上可接受的载体或赋形剂包括稳定剂、稀释剂、表面活性剂、润滑剂、抗氧化剂、粘合剂、着色剂、填充剂、乳化剂等。
无菌可注射组合物可按照本领域已知的技术使用适合的分散剂或润湿剂和助悬剂来配制。可以使用的药学上可接受的载体和溶剂包括水、甘露醇、氯化钠溶液等。
可以改变本发明的药物组合物中活性成分的实际剂量水平以获得对特定患者、组合物和施用方式而言可以有效实现所需治疗响应、对患者无毒的活性成分的量。所选择的的剂量水平取决于多种因素，包括所用的具体的本发明的化合物或其盐的活性、施用途径、施用时间、所用的具体组合物的排泄速率、治疗的持续时间、与所用的具体组合物组合使用的其它药物、化合物和/或材料、所治疗的患者的年龄、性别、体重、一般健康状况和既往病史以及医学领域中公知的类似因素。
本发明另一目的在于提供通式I、通式II的化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体在制备用于治疗或预防由GSPT1、IKZF1、IKZF2、IKZF3、CK1α、N-MYC或C-MYC蛋白突变、表达失衡、变构与功能异常相关疾病的药物中的用途。
所述相关疾病为癌症、衰老、免疫性疾病、神经性疾病；其中，所述癌症选自急性髓白血病、肝癌、急性淋巴细胞白血病、膀胱癌、骨癌、脑癌、乳腺癌、宫颈癌、绒毛膜癌、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)、结肠癌、食道癌、胆囊癌、胃癌、胃肠道间质瘤、头颈癌、霍奇金淋巴瘤、喉癌、白血病、肺癌、黑色素瘤、间皮瘤、多发性骨髓瘤、卵巢癌、胰腺癌、前列腺癌、直肠癌、肾癌、肉瘤、皮肤癌、小细胞肺癌、睾丸癌、咽喉癌、甲状腺癌或子宫癌。
本发明还提供了通式I、通式II的化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体在制备GSPT1降解剂中的用途。
 有益效果：
与临床药物CC-90009相比，本发明的苯并六元杂环化合物如A、B、C系列对GSPT1蛋白的降解效果、AML细胞的抗增殖抑制活性显著提高。其中，200nM化合物A-10在KG-1细胞时给药4小时，对GSPT1蛋白的降解百分数高于80％。化合物A-1、A-3、A-10的体外大鼠肝微粒体代谢半衰期与临床药物CC-90009相比有着约2-5倍的优势，具有优秀的代谢稳定性。药代动力学研究中，化合物A-1静脉注射给药时，体内清除率小于临床药物CC-90009，半衰期与其接近，在体内具有较好的代谢稳定性，初始血药浓度以及血浆药物暴露量与CC-90009相比具有较大优势。化合物A-10静脉注射给药的清除率是CC-90009的三分之一，且半衰期长于CC-90009，初始血药浓度远高于CC-90009，血浆药物暴露量接近其四倍，并且口服给药的暴露量接近CC-90009三倍，具有较高的达峰高度和较小的达峰时间，揭示化合物A-10吸收更快。化合物A-42清除率更低，具有更高的静脉和口服血浆药物暴露量，分别为CC-90009的十四倍和四倍。此外，异种移植小鼠模型的药效学研究表明，化合物A-10、A-42具有较好的体内抗肿瘤作用，其中A-10体内抗肿瘤作用明显优于临床分子CC-90009，A-42体内抗肿瘤作用与临床分子MRT-2359水平相当。因此，本发明化合物可以用于制备治疗或预防由GSPT1、IKZF1、IKZF2、IKZF3、CK1α、N-MYC或C-MYC蛋白突变、表达失衡、变构与功能异常相关疾病的药物；可以用于制备GSPT1降解剂。
除非有相反陈述，在说明书和权利要求书中使用的术语具有下述含义。
术语“异构体”包括给定结构的对映异构形式、非对映异构形式和几何(或构象)异构形式。例如，本申请包括每个不对称中心的R和S构型、Z和E双键异构体、Z和E构象异构体、单一立体化学异构体及对映异构体、非对映异构体和几何(或构象)异构体混合物。
术语“药学上可接受的盐”指，诸如其酸加成盐和/或碱盐。合适的酸加成盐由酸形成，其形成无毒盐，例如盐酸盐/氯化物。合适的碱盐由碱形成，其形成无毒盐，例如钙盐和钠盐。还可形成酸和碱的半盐，例如半硫酸盐和半钙盐。
术语“治疗有效量”是指本申请化合物的以下量，其(i)治疗具体的疾病、病症或障碍；(ii)减轻、缓解或消除具体的疾病、病症或障碍的一种或多种症状；或(iii)预防或延迟本申请所述具体的疾病、病症或障碍的一种或多种症状的发作。
术语“药学上可接受的载体或赋形剂”是指不破坏用其配制的化合物的药理活性的无毒载体、辅料或媒介物。
术语“烷基”指饱和脂肪族烃基团，本发明优选含有1至6个碳原子的烷基。含有1至6个碳原子的低级烷基的非限制性实施例包括甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基等。
术语“烯基”指具有至少一个碳-碳双键的脂族烃，包括具有至少一个碳-碳双键的直链和支链。本发明烯基基团优选具有2至6个碳原子。例如，术语“C2-6烯基”包括2至6个碳原子的直链或支链不饱和基团(具有至少一个碳-碳双键)，包括但不限于乙烯基、1-丙烯基、2-丙烯基(烯丙基)、异丙烯基、2-甲基-1-丙烯基、1-丁烯基、2-丁烯基等。
术语“炔基”指具有至少一个碳-碳三键的脂族烃，包括具有至少一个碳-碳三键的直链和支链。本发明炔基基团具有2至6个碳原子。例如，“C2-6炔基”包括具有2至6个碳原子的如上定义的直链或支链烃链炔基基团。
术语“烷氧基”指-O-(烷基)和-O-(非取代的环烷基)，其中烷基如上述定义。烷氧基的非限制性实例包括：甲氧基、乙氧基、丙氧基、丁氧基、环丙氧基、环丁氧基、环戊氧基、环己氧基。
术语“环烷基”指饱和或部分不饱和单环或多环环状烃取代基，本发明环烷基优选包含3至10个碳原子。单环环烷基的非限制性实例包括环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基、环庚三烯基、环辛基等；多环环烷基包括螺环、稠环和桥环的环烷基。
术语“螺环烷基”指5至20元的单环之间共用一个碳原子(称螺原子)的多环基团，其可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。优选为6至14元，更优选为7至10元(例如7、8、9或10元)。根据环与环之间共用螺原子的数目将螺环烷基分为单螺环烷基、双螺环烷基或多螺环烷基，优选为单螺环烷基和双螺环烷基，更优选为4元/4元、4元/5元、4元/6元、5元/5元或5元/6元单螺环烷基。
术语“稠环烷基”指5至20元，系统中的每个环与体系中的其他环共享毗邻的一对碳原子的全碳多环基团，其中一个或多个环可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。优选为6至14元，更优选为7至10元。根据组成环的数目可以分为双环、三环、四环或多环稠环烷基，优选为双环或三环，更优选为5元/5元或5元/6元双环烷基。
术语“桥环烷基”指5至20元，任意两个环共用两个不直接连接的碳原子的全碳多环基团，其可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。优选为6至14元，更优选为7至10元。根据组成环的数目可以分为双环、三环、四环或多环桥环烷基，优选为双环、三环或四环，更优选为双环或三环。
术语“杂环基”指饱和或部分不饱和单环或多环环状烃取代基，其包含3至20个环原子，其中一个或多个环原子为选自氮、氧或S(O)m(其中m是整数0至2)的杂原子，但不包括-O-O-、-O-S-或-S-S-的环部分，其余环原子为碳。优选包含3至10个环原子，其中1～4个是杂原子；最优选包含3至8个环原子，其中1～3个是杂原子；最优选包含5至6个环原子，其中1～2或1～3个是杂原子。单环杂环基的非限制性实例包括吡咯烷基、咪唑烷基、四氢呋喃基、四氢吡喃基、四氢噻吩基、二氢咪唑基、二氢呋喃基、二氢吡唑基、二氢吡咯基、哌啶基、哌嗪基、吗啉基、硫代吗啉基、高哌嗪基等，优选四氢吡喃基、哌啶基、吡咯烷基。多环杂环基包括螺杂环基、稠杂环基和桥杂环基。
术语“螺杂环基”指5至20元的单环之间共用一个原子(称螺原子)的多环杂环基团，其中一个或多个环原子为选自氮、氧或S(O)m(其中m是整数0至2)的杂原子，其余环原子为碳。其可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。优选为6至14元，更优选为7至10元。根据环与环之间共用螺原子的数目将螺杂环基分为单螺杂环基、双螺杂环基或多螺杂环基，优选为单螺杂环基和双螺杂环基。更优选为4元/4元、4元/5元、4元/6元、5元/5元或5元/6元单螺杂环基。
术语“稠杂环基””指5至20元，系统中的每个环与体系中的其他环共享毗邻的一对原子的多环杂环基团，一个或多个环可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统，其中一个或多个环原子为选自氮、氧或S(O)m(其中m是整数0至2)的杂原子，其余环原子为碳。优选为6至14元，更优选为7至10元。根据组成环的数目可以分为双环、三环、四环或多环稠杂环基，优选为双环或三环，更优选为5元/5元或5元/6元双环稠杂环基。
术语“桥杂环基”指5至14元，任意两个环共用两个不直接连接的原子的多环杂环基团，其可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统，其中一个或多个环原子为选自氮、氧或S(O)m(其中m是整数0至2)的杂原子，其余环原子为碳。优选为6至14元，更优选为7至10元。根据组成环的数目可以分为双环、三环、四环或多环桥杂环基，优选为双环、三环或四环，更优选为双环或三环。
所述杂环基包括如上所述的杂环基(包括单环、螺杂环、稠杂环和桥杂环)稠合于芳基、杂芳基或环烷基环上，其中与母体结构连接在一起的环为杂环基，其非限制性实例包括：
术语“芳基”指具有共轭的π电子体系的6至14元全碳单环或稠合多环(也就是共享毗邻碳原子对的环)基团，优选为6至10元，例如苯基和萘基。
术语“杂芳基”指包含1至4个杂原子、5至14个环原子的杂芳族体系，其中杂原子选自氧、硫和氮。杂芳基优选为5至10元，含1至3个杂原子；更优选为5元或6元，含1至2个杂原子；优选例如咪唑基、呋喃基、噻吩基、噻唑基、吡唑基、噁唑基、吡咯基、四唑基、吡啶基、嘧啶基、噻二唑、吡嗪基和哒嗪基等。
所述杂芳基包括如上所述的杂芳基稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环，其非限制性实例包括：
术语“羟烷基”指被羟基取代的烷基，其中烷基如上所定义。
术语“卤代烷基”指烷基被一个或多个卤素取代，其中烷基如上所定义。
术语“卤代烷氧基”指烷氧基被一个或多个卤素取代，其中烷氧基如上所定义。
 附图说明
图1为本发明化合物A-10对AML细胞中的GSPT1降解作用呈现浓度依赖。其中，图1A为化合物A-10与临床药物CC-90009在KG-1细胞中对GSPT1的降解作用；图1B为A图中的GSPT1蛋白定量结果；图1C为化合物A-10与临床药物CC-90009在MV4-11细胞中对GSPT1的降解作用；图1D为C图中的GSPT1蛋白定量结果。
图2为本发明化合物A-10对AML细胞中的GSPT1降解作用呈现时间依赖。
图3为本发明化合物A-1、A-3、A-10、A-42、B-1以及临床药物CC-90009在体外大鼠肝微粒体中的代谢稳定性研究结果。
图4为本发明化合物A-1、A-10、A-42在大鼠体内药代动力学研究结果；其中，图4A、4B分别表示CC-90009静脉和口服给药的药时曲线结果；图4C、4D分别表示A-1静脉和口服给药的药时曲线结果；图4E、4F分别表示A-10静脉和口服给药的药时曲线结果；图4G、4H分别表示A-42静脉和口服给药的药时曲线结果。
图5为本发明化合物A-10在MV411异种移植小鼠模型中的体内药效学研究；其中，图5A为各组小鼠体重；
图5B为各组小鼠肿瘤生长曲线；图5C为各组小鼠肿瘤质量；图5D为各组小鼠肿瘤组织解剖。
图6为本发明化合物A-42在MV411异种移植小鼠模型中的体内药效学研究；其中，图6A为各组小鼠体重；
图6B为各组小鼠肿瘤生长曲线。
[根据细则91更正 16.05.2024]
图7为本发明化合物A-10在22RV1异种移植小鼠模型中的体内药效学研究；其中，图7A为各组小鼠体重；图7B为各组小鼠肿瘤生长曲线；图7C为各组小鼠肿瘤质量；图7D为各组小鼠肿瘤组织解剖。
[根据细则91更正 16.05.2024]
 具体实施方式
下面通过具体实施例对本发明技术方案进行详细说明，但是本发明的保护范围不局限于所述实施例。关键中间体-1的合成：
步骤1：将4-溴-2-甲基苯甲酸甲酯(10.00g，43.86mmol)溶解于200mL的四氯化碳中，依次加入N-溴代丁二酰亚胺(15.61g，87.73mmol)和偶氮二异丁腈(0.72g，4.38mmol)，加毕，升温至回流搅拌12小时。经TLC监测反应完全，冷却至室温后抽滤，滤液真空浓缩，残余物使用硅胶色谱柱(DCM：MeOH＝40：1，v/v)纯化，得到白色固体化合物2：4-溴-2-(溴甲基)苯甲酸甲酯(8.32g，收率62.0％)。HRMS(ESI+):计算值C9H9Br2O2(M+H)+,306.8964；实测值306.8958。
步骤2：将4-溴-2-(溴甲基)苯甲酸甲酯(8.00g，26.15mmol)溶解于60mL的N,N-二甲基甲酰胺中，依次加入3-氨基哌啶-2,6-二酮盐酸盐(5.15g，31.38mmol)和DIPEA(8.45g，65.38mmol)，加毕，于氮气保护下升温至90℃搅拌4小时。经TLC监测反应完全，将反应液倒入200mL水中，析出浅蓝色固体，静置使其完全析出，抽滤，滤饼真空干燥得到浅蓝色固体化合物3：3-(5-溴-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(5.89g，收率70.2％)。HRMS(ESI+):计算值C13H12BrN2O3(M+H)+,323.0026；实测值323.0030。
步骤3：将3-(5-溴-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(5.00g，15.53mmol)溶于50mL的干燥N,N-二甲基甲酰胺中，依次加入1,1’-双(二苯基磷)二茂铁(0.40g，0.71mmol)、三(二亚苄基丙酮)二钯(0.39g，0.43mmol)和氰化锌(1.88g，16.00mmol)，加毕，于氮气保护下升温至90℃搅拌4小时。经TLC监测反应完全，抽滤，将滤液倒入150mL水中，析出绿色固体，静置使其完全析出，抽滤，滤饼真空干燥得到浅绿色固体化合物4：2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-甲腈(3.84g，收率91.9％)。HRMS(ESI+):计算值C14H12N3O3(M+H)+,270.0873；实测值270.0882。
步骤4：将2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-甲腈(3.50g，13.00mmol)溶于40mL的N,N-二甲基甲酰胺中，依次加入二碳酸二叔丁酯(5.67g，26.00mmol)和15mL的雷尼镍，加毕，于氢气充分置换下升温至50℃搅拌5小时。经TLC监测反应完全，抽滤，滤饼使用乙酸乙酯洗涤，滤液倒入120mL水中，使用乙酸乙酯萃取，合并有机相，并依次使用水和饱和食盐水洗涤，无水硫酸钠干燥后真空浓缩得到棕黄色固体化合物5：((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)氨基甲酸叔丁酯(3.20g，收率65.9％)，该产物无需进一步纯化可直接用于下一步。HRMS(ESI+):计算值C19H24N3O5(M+H)+,374.1710；实测值374.1721。
步骤5：将((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)氨基甲酸叔丁酯(3.00g，8.04mmol)溶于30mL的盐酸-二氧六环溶液(4M)中，室温搅拌3小时。经TLC监测反应完全，抽滤，滤饼真空干燥得到淡黄色固体，即为中间体-1：3-(5-氨甲基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮盐酸盐(1.80g，收率72.4％)，该产物无需进一步纯化可直接用于下一步。HRMS(ESI+):计算值C14H16N3O3(M+H)+,274.1186；实测值274.1184。
关键中间体-2的合成：
步骤1：将2-羟基苯甲醛(2.00g，16.39mmol)溶于15mL的丙烯腈中，加入1,4-二叠氮双环[2.2.2]辛烷(1.84g，16.39mmol)，加毕，于氮气保护下升温至回流搅拌3小时。经TLC监测反应完全，将反应液倒入30mL的10％氢氧化钠水溶液中，使用乙酸乙酯萃取，合并有机相，并使用饱和食盐水洗涤，无水硫酸钠干燥后，真空浓缩得到黄色结晶状固体化合物7：2H-色烯-3-腈(2.30g，收率90.2％)，该产物无需进一步纯化可直接用于下一步。HRMS(ESI+):计算值C10H8NO(M+H)+,158.0600；实测值158.0621。
步骤2：将2H-色烯-3-腈(2.30g，14.73mmol)溶于40mL的10％氢氧化钠水溶液中，升温至60℃搅拌10小时。经TLC监测反应完全，添加稀盐酸(2M)调节pH至4-5，析出白色固体，静置使其完全析出，抽滤，干燥得黄色固体化合物8：2H-色烯-3-羧酸(1.65g，收率63.7％)。HRMS(ESI-):计算值C10H7O3(M-H)-,175.0401；实测值175.0400。
步骤3：将2H-色烯-3-羧酸(1.50g，8.52mmol)溶于20mL的甲醇中，加入10％ Pd/C(1.00g)，加毕，室温搅拌8小时。经TLC监测反应完全，抽滤，滤饼使用甲醇洗涤，滤液真空浓缩得到棕黄色固体，即为中间体-2：苯并二氢吡喃-3-羧酸(0.76g，收率50.2％)。HRMS(ESI-):计算值C10H9O3(M-H)-,177.0557；实测值177.0545。
关键中间体-3的合成：
步骤1：将4-溴-3-氟苯甲酸(9，15.00g，68.49mmol)溶解于150mL的无水四氢呋喃中，氮气保护。降温到-70℃，滴加二异丙基氨基锂(72mL，143.8mmol)，滴加完毕后继续反应1小时，滴加碘甲烷(29.16g，205.5mmol)，移至室温反应12小时。经TLC监测反应完全，向反应液中逐滴加入150mL饱和氯化铵溶液。使用乙酸乙酯萃取，合并有机相，并使用饱和食盐水洗涤，无水硫酸钠干燥后，真空浓缩得到黄色结晶状固体化合物10：4-溴-3-氟-2-甲基苯甲酸(9.67g，收率60.6％)，该产物无需进一步纯化可直接用于下一步。HRMS(ESI-)：计算值C8H5BrFO2(M-H)-，230.9462；实测值230.9455。
步骤2：将4-溴-3-氟-2-甲基苯甲酸(10，9.67g，41.50mmol)溶解于80mL的N,N-二甲基甲酰胺中，依次加入碳酸钾(11.47g，83.01mmol)和碘甲烷(8.84g，62.26mmol)，加毕，室温反应2小时。经TLC监测反应完全，抽滤，滤液倒入300mL水中，使用乙酸乙酯萃取，合并有机相，并依次使用水、饱和食盐水洗涤，无水硫酸钠干燥后，真空浓缩得到黄色油状物11：4-溴-3-氟-2-甲基苯甲酸甲酯(9.29g，收率90.6％)，该产物无需进一步纯化可直接用于下一步。HRMS(ESI+)：计算值C9H9BrFO2(M+H)+，246.9764；实测值246.9752。
步骤3：将4-溴-3-氟-2-甲基苯甲酸甲酯(11，9.29g，37.60mmol)溶解于150mL的氯仿中，依次加入N-溴代丁二酰亚胺(16.73g，94.00mmol)和偶氮二异丁腈(1.85g，11.28mmol)，加毕，升温至回流反应10小时。经TLC监测反应完全，抽滤，滤液真空浓缩，加入石油醚打浆得到白色固体12：4-溴-2-(溴甲基)-3-氟苯甲酸甲酯(10.36g，收率84.5％)。HRMS(ESI+)：计算值C9H8Br2FO2(M+H)+，324.8870；实测值324.8812。
步骤4-步骤7：采用原料4-溴-2-(溴甲基)-3-氟苯甲酸甲酯参照中间体-1的合成方法制备，得到白色固体中间体-3：3-(5-(氨甲基)-4-氟-1-氧代异吲哚啉-2-基)哌啶-2，6-二酮盐酸盐。HRMS(ESI+)：计算值C14H15FN3O3(M+H)+，292.1092；实测值292.1077。
临床药物CC-90009的合成：
将3-(5-氨甲基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮盐酸盐(0.10g，0.32mmol)溶解于5mL的N,N-二甲基甲酰胺中，依次加入2-(4-氯苯基)-2,2-二氟乙酸(0.08g，0.39mmol)、HATU(0.19g，0.49mmol)，加毕，室温搅拌1小时。加入DIPEA(0.13g，0.97mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入30mL冰水中，析出棕色固体，静置使其完全析出，抽滤，将滤饼使用硅胶色谱柱(二氯甲烷：甲醇＝50：1,v/v)纯化，得到白色固体CC-90009(0.10g，收率68.0％，纯度99.37％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),9.70(t,J＝6.1Hz,1H),7.68(d,J＝7.8Hz,1H),7.62(s,4H),7.41(s,1H),7.37(d,J＝7.8Hz,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.53-4.22(m,4H),3.02-2.83(m,1H),2.60(d,J＝17.3Hz,1H),2.47-2.30(m,1H),2.06-1.94(m,1H)。HRMS(ESI+):计算值C22H19ClF2N3O4(M+H)+,462.1027；实测值462.1025。
实施例1：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2H-色烯-3-甲酰胺的合成(化合物A-1)
将3-(5-氨甲基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮盐酸盐(中间体-1，0.10g，0.32mmol)溶解于5mL的N,N-二甲基甲酰胺中，依次加入2H-色烯-3-羧酸(化合物8，0.07g，0.39mmol)、HATU(0.19g，0.49mmol)，加毕，室温搅拌1小时。加入DIPEA(0.13g，0.97mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入30mL冰水中，析出棕色固体，静置使其完全析出，抽滤，将滤饼使用硅胶色谱柱(二氯甲烷：甲醇＝50：1,v/v)纯化，得到白色固体，即化合物A-1(0.10g，收率71.9％，纯度98.95％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.90(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.45(d,J＝7.3Hz,1H),7.33(s,1H),7.29-7.18(m,2H),6.96(t,J＝7.4Hz,1H),6.85(d,J＝8.1Hz,1H),5.11(dd,J＝13.3,5.1Hz,1H),4.94(s,2H),4.56-4.24(m,4H),3.01-2.81(m,1H),2.60(d,J＝17.5Hz,1H),2.49-2.28(m,1H),2.08-1.93(m,1H)。HRMS(ESI+):计算值C24H22N3O5(M+H)+,432.1554；实测值432.1539。
实施例2：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-5-氟-2H-色烯-3-甲酰胺的合成(化合物A-2)
5-氟-2H-色烯-3-羧酸(16)的合成：采用原料6-氟水杨醛参照中间体-2的合成方法制备，两步收率共计52.4％。HRMS(ESI-):计算值C10H6FO3(M-H)-,193.0306；实测值193.0318。
化合物A-2的合成：采用原料5-氟-2H-色烯-3-羧酸参照实施例1化合物A-1的合成方法制备，得到白色固体，即为化合物A-2(0.08g，收率55.9％，纯度95.93％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.96(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.45(d,J＝7.8Hz,1H),7.37(s,1H),7.26-7.16(m,1H),7.08(d,J＝7.6Hz,1H),7.00-6.88(m,1H),5.11(dd,J＝13.3,5.0Hz,1H),5.02(s,2H),4.55-4.23(m,4H),3.02-2.83(m,1H),2.60(d,J＝17.3Hz,1H),2.47-2.30(m,1H),2.06-1.93(m,1H)。HRMS(ESI+):计算值C24H21FN3O5(M+H)+,450.1460；实测值450.1436。
实施例3：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氟-2H-色烯-3-甲酰胺的合成(化合物A-3)
6-氟-2H-色烯-3-羧酸(17)的合成：采用原料5-氟水杨醛参照中间体-2的合成方法制备，两步收率共计62.1％。HRMS(ESI-):计算值C10H6FO3(M-H)-,193.0306；实测值193.0310。
化合物A-3的合成：采用原料6-氟-2H-色烯-3-羧酸参照实施例1化合物A-1的合成方法制备，得白色固体，即为A-3(0.12g，收率83.9％，纯度98.72％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.02(s,1H),8.97(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.52(s,1H),7.45(d,J＝7.2Hz,1H),7.30(s,1H),7.18-7.12(m,1H),7.12-7.03(m,1H),6.89(dd,J＝8.8,4.6Hz,1H),5.12(dd,J＝13.3,5.0Hz,1H),4.94(s,2H),4.54-4.24(m,4H),3.00-2.85(m,1H),2.60(d,J＝17.9Hz,1H),2.47-2.29(m,1H),2.06-1.93(m,1H)。HRMS(ESI+):计算值C24H21FN3O5(M+H)+,450.1460；实测值450.1428。
实施例4：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-7-氟-2H-色烯-3-甲酰胺的合成(化合物A-4)
7-氟-2H-色烯-3-羧酸(18)的合成：采用原料4-氟水杨醛参照中间体-2的合成方法制备，两步收率共计56.8％。HRMS(ESI-):计算值C10H6FO3(M-H)-,193.0306；实测值193.0321。
化合物A-4的合成：采用原料7-氟-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体，即为化合物A-4(0.08g，收率55.9％，纯度95.38％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.02(s,1H),8.92(t,J＝6.1Hz,1H),7.70(d,J＝7.8Hz,1H),7.52(s,1H),7.45(d,J＝7.9Hz,1H),7.34(s,1H),7.29(t,J＝7.4Hz,1H),6.86-6.74(m,2H),5.12(dd,J＝13.2,5.0Hz,1H),4.98(s,2H),4.55-4.24(m,4H),3.02-2.84(m,1H),2.60(d,J＝15.9Hz,1H),2.47-2.30(m,1H),2.07-1.94(m,1H)。HRMS(ESI+):计算值C24H21FN3O5(M+H)+,450.1460；实测值450.1454。
实施例5：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-8-氟-2H-色烯-3-甲酰胺的合成(化合物A-5)
8-氟-2H-色烯-3-羧酸(19)的合成：采用原料3-氟水杨醛参照中间体-2的合成方法制备，两步收率共计59.2％。HRMS(ESI-):计算值C10H6FO3(M-H)-,193.0306；实测值193.0311。
化合物A-5的合成：采用原料8-氟-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-5(0.09g，收率62.9％，纯度99.89％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),9.07(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.51(d,J＝7.3Hz,2H),7.45(d,J＝7.9Hz,1H),7.35-7.21(m,1H),6.84(t,J＝8.8Hz,1H),6.74(d,J＝8.2Hz,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.97(s,2H),4.57-4.24(m,4H),3.02-2.85(m,1H),2.60(d,J＝17.0Hz,1H),2.47-2.33(m,1H),2.10-1.92(m,1H)。HRMS(ESI+):计算值C24H21FN3O5(M+H)+,450.1460；实测值450.1432。
实施例6：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氯-2H-色烯-3-甲酰胺的合成(化合物A-6)
6-氯-2H-色烯-3-羧酸(20)的合成：采用原料5-氯水杨醛参照中间体-2的合成方法制备，两步收率共计62.5％。HRMS(ESI-):计算值C10H6ClO3(M-H)-,209.0011；实测值209.0023。
化合物A-6的合成：采用原料6-氯-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-6(0.06g，收率40.5％，纯度97.87％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.93(t,J＝6.1Hz,1H),7.70(d,J＝7.7Hz,1H),7.53(s,1H),7.45(d,J＝7.8Hz,1H),7.32(s,1H),7.28(s,1H),7.25(s,1H),6.88(d,J＝8.6Hz,1H),5.11(dd,J＝13.4,5.0Hz,1H),4.97(s,2H),4.57-4.26(m,4H),3.04-2.82(m,1H),2.60(d,J＝17.3Hz,1H),2.44-2.29(m,1H),2.08-1.91(m,1H)。HRMS(ESI+):计算值C24H21ClN3O5(M+H)+,466.1164；实测值466.1169。
实施例7：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-甲基-2H-色烯-3-甲酰胺的合成(化合物A-7)
6-甲基-2H-色烯-3-羧酸(21)的合成：采用原料5-甲基水杨醛参照中间体-2的合成方法制备，两步收率共计57.6％。HRMS(ESI-):计算值C11H9O3(M-H)-,189.0557；实测值189.0548.
化合物A-7的合成：采用原料6-甲基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-7(0.07g，收率49.3％，纯度98.76％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.89(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.52(s,1H),7.45(d,J＝7.8Hz,1H),7.29(s,1H),7.04(d,J＝8.1Hz,2H),6.75(d,J＝7.9Hz,1H),5.11(dd,J＝13.2,5.0Hz,1H),4.90(s,2H),4.56-4.24(m,4H),2.99-2.83(m,1H),2.60(d,J＝17.0Hz,1H),2.48-2.31(m,1H),2.23(s,3H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C25H24N3O5(M+H)+,446.1711；实测值446.1706。
实施例8：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-甲氧基-2H-色烯-3-甲酰胺的合成(化合物A-8)
6-甲氧基-2H-色烯-3-羧酸(22)的合成：采用原料5-甲氧基水杨醛参照中间体-2的合成方法制备，两步收率共计58.6％。HRMS(ESI-):计算值C11H9O4(M-H)-,205.0506；实测值205.0511。
化合物A-8的合成：采用原料6-甲氧基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-8(0.06g，收率40.8％，纯度99.05％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.88(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.52(s,1H),7.45(d,J＝7.8Hz,1H),7.30(s,1H),6.88-6.75(m,3H),5.11(dd,J＝13.3,5.1Hz,1H),4.87(s,2H),4.56-4.25(m,4H),3.71(s,3H),3.00-2.83(m,1H),2.60(d,J＝17.1Hz,1H),2.47-2.30(m,1H),2.07-1.94(m,1H)。HRMS(ESI+):计算值C25H24N3O6(M+H)+,462.1660；实测值462.1659。
实施例9：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-5-甲氧基-2H-色烯-3-甲酰胺的合成(化合物A-9)
5-甲氧基-2H-色烯-3-羧酸(23)的合成：采用原料6-甲氧基水杨醛参照中间体-2的合成方法制备，两步收率共计62.5％。HRMS(ESI-):计算值C11H9O4(M-H)-,205.0506；实测值205.0515。
化合物A-9的合成：采用原料5-甲氧基-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-9(0.07g，收率47.6％，纯度99.23％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.97(t,J＝6.0Hz,1H),7.69(d,J＝7.8Hz,1H),7.53(d,J＝11.1Hz,2H),7.44(d,J＝7.9Hz,1H),7.21(t,J＝8.2Hz,1H),6.63(d,J＝8.4Hz,1H),6.49(d,J＝8.2Hz,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.86(s,2H),4.53-4.24(m,4H),3.83(s,3H),2.99-2.83(m,1H),2.60(d,J＝17.1Hz,1H),2.49-2.28(m,1H),2.07-1.89(m,1H)。HRMS(ESI+):计算值C25H24N3O6(M+H)+,462.1660；实测值462.1639。
实施例10：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-三氟甲氧基-2H-色烯-3-甲酰胺的合成(化合物A-10)
6-三氟甲氧基-2H-色烯-3-羧酸(24)的合成：采用原料5-三氟甲氧基水杨醛参照中间体-2的合成方法制备，两步收率共计62.3％。HRMS(ESI-):计算值C11H6F3O4(M-H)-,259.0224；实测值259.0228。
化合物A-10的合成：采用原料6-三氟甲氧基-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-10(0.12g，收率72.7％，纯度99.89％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),8.96(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.45(d,J＝7.8Hz,1H),7.33(s,1H),7.31(d,J＝3.1Hz,1H),7.28-7.20(m,1H),6.96(d,J＝8.8Hz,1H),5.12(dd,J＝13.3,5.1Hz,1H),5.00(s,2H),4.56-4.25(m,4H),3.00-2.81(m,1H),2.60(d,J＝17.6Hz,1H),2.48-2.30(m,1H),2.07-1.94(m,1H)。HRMS(ESI+):计算值C24H21F3N3O6(M+H)+,516.1377；实测值516.1370。
实施例11：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氰基-2H-色烯-3-甲酰胺的合成(化合物A-11)
步骤1：将3-甲酰基-4-羟基苯腈(25，1.00g，6.80mmol)溶于20mL的1,4-二氧六环中，依次加入碳酸钾(0.94g，6.80mmol)、丙烯醛(0.76g，13.60mmol)，加毕，升温至回流搅拌5小时。经TLC监测反应完全，将反应液倒入20mL水中，使用乙酸乙酯萃取，合并有机相，并依次使用2M氢氧化钠溶液和饱和食盐水洗涤，无水硫酸钠干燥后，真空浓缩，将残余物使用硅胶色谱柱(石油醚：乙酸乙酯＝10：1，v/v)纯化，得到黄色固体26：3-甲酰基-2H-色烯-6-腈(1.03g，收率82.0％)。HRMS(ESI+):计算值C11H8NO2(M+H)+,186.0550；实测值186.0548。
步骤2：3-甲酰基-2H-色烯-6-腈(1.03g，5.57mmol)溶于20mL的四氢呋喃中，逐滴加入溶于15mL水中的亚氯酸钠(5.03g，55.66mmol)和磷酸二氢钠(6.68g，55.66mmol)，加入2-甲基-2丁烯(4.27g，60.81mmol)。加毕，室温搅拌4小时。经TLC监测反应完全，将反应液真空浓缩后用30mL水稀释，水层用二氯甲烷萃取，合并有机相，并使用饱和食盐水洗涤，无水硫酸钠干燥后，真空浓缩得到黄色油状物27：6-氰基-2H-色烯-3-羧酸(0.90g，收率80.0％)，该产物无需进一步纯化可直接用于下一步。HRMS(ESI+):计算值C11H8NO3(M+H)+,202.0499；实测值202.0496。
步骤3：采用原料6-氰基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得到白色固体A-11：(0.08g，收率54.8％，纯度96.95％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),8.92(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.52(s,1H),7.42(d,J＝7.3Hz,1H),7.30(s,1H),7.23(t,J＝8.2Hz,1H),6.96(t,J＝7.4Hz,1H),6.85(d,J＝8.1Hz,1H),5.12(dd,J＝13.3,5.1Hz,1H),4.94(s,2H),4.56-4.24(m,4H),2.99-2.80(m,1H),2.59(d,J＝17.5Hz,1H),2.47-2.28(m,1H),2.06-1.95(m,1H)。HRMS(ESI+):计算值C25H21N4O5(M+H)+,457.1507；实测值457.1502。
实施例12：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氨基-2H-色烯-3-甲酰胺的合成(化合物A-12)
步骤1：将N-乙酰对氨基酚(28，10.00g，66.20mmol)溶于40mL的三氟乙酸中，于冰浴下将六亚甲基四胺(36.56g，0.26mol)分批加入到反应液中，加毕，升温至70℃搅拌5小时。经TLC监测反应完全，反应液冷却至室温后，倒入200mL水中，使用乙酸乙酯萃取，合并有机相并使用饱和食盐水洗涤，无水硫酸钠干燥后真空浓缩，将残余物使用硅胶色谱柱(石油醚：乙酸乙酯＝1：1，v/v)纯化，得到白色固体29：N-(3-甲酰基-4-羟基苯基)乙酰胺(2.03g，收率17.1％)。HRMS(ESI+):计算值C9H10NO3(M+H)+,180.0655；实测值180.0638。
步骤2：将N-(3-甲酰基-4-羟基苯基)乙酰胺(2.03g，11.34mmol)溶于乙醇(30mL)和稀盐酸(6M，10mL)中，升温至回流搅拌8小时。经TLC监测反应完全，将反应液真空浓缩，用乙醇打浆得到黑色固体30：5-氨基-2-羟基苯甲醛盐酸盐(1.93g，收率98.5％)。HRMS(ESI+):计算值C7H8NO2(M+H)+,138.0550；实测值138.0545。
步骤3：将5-氨基-2-羟基苯甲醛盐酸盐(1.93g，11.15mmol)溶于30mL的四氢呋喃中，依次加入三乙胺(1.13g，11.15mmol)和二碳酸二叔丁酯(2.92g，13.39mmol)，加毕，室温搅拌16小时。经TLC监测反应完全，将反应液真空浓缩，将残余物使用硅胶色谱柱(石油醚：乙酸乙酯＝10：1，v/v)纯化，得到白色固体31：(3-甲酰基-4-羟基苯基)氨基甲酸叔丁酯(2.61g，收率98.9％)。HRMS(ESI+):计算值C12H16NO4(M+H)+,238.1074；实测值238.1065。
步骤4-步骤6：采用原料(3-甲酰基-4-羟基苯基)氨基甲酸叔丁酯参照中间体-2和化合物A-1的合成方法制备，得到白色固体34：((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)氨基甲酰基)-2H-色烯-6-基)氨基甲酸叔丁酯(1.21g，收率20.1％)。HRMS(ESI+):计算值C29H31N4O7(M+H)+,547.2187；实测值547.2188。
步骤7：将((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)氨基甲酰基)-2H-色烯-6-基)氨基甲酸叔丁酯(1.21g，2.22mmol)溶于30mL的盐酸-二氧六环溶液(4M)中，室温搅拌2小时。经TLC监测反应完全，将反应液真空浓缩后使用饱和碳酸氢钠溶液将残余物调至弱碱性，抽滤，滤饼用少量石油醚洗涤，真空干燥得白色固体A-12(0.99g，收率95.0％，纯度98.70％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.02(s,1H),8.99(t,J＝6.0Hz,1H),7.69(d,J＝7.8Hz,1H),7.52(s,1H),7.44(d,J＝7.3Hz,1H),7.31(s,1H),7.24(t,J＝8.1Hz,1H),6.96(t,J＝7.4Hz,1H),6.85(d,J＝8.1Hz,1H),5.51(s,2H),5.12(dd,J＝13.3,5.1Hz,1H),4.93(s,2H),4.50-4.24(m,4H),2.99-2.81(m,1H),2.61(d,J＝17.5Hz,1H),2.47-2.28(m,1H),2.06-1.95(m,1H)。HRMS(ESI+):计算值C24H23N4O5(M+H)+,447.1663；实测值447.1660。
实施例13：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-(二甲氨基)-2H-色烯-3-甲酰胺的合成(化合物A-13)
6-(二甲氨基)-2H-色烯-3-羧酸(35)的合成：采用原料5-二甲氨基水杨醛参照中间体-2的合成方法制备，两步收率共计48.6％。HRMS(ESI-):计算值C12H12NO3(M-H)-,218.0823；实测值218.0821。
化合物A-13的合成：采用原料6-(二甲氨基)-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得灰白色固体A-13(0.06g，收率40.0％，纯度97.13％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.02(s,1H),8.99(t,J＝6.0Hz,1H),7.67(d,J＝7.7Hz,1H),7.56(s,1H),7.45(d,J＝7.3Hz,1H),7.31(d,J＝8.1,1H),6.67-6.50(m,3H),5.12(dd,J＝13.3,5.1Hz,1H),4.93(s,2H),4.50-4.24(m,4H),3.12(s,6H),2.98-2.80(m,1H),2.60(d,J＝17.5Hz,1H),2.48-2.30(m,1H),2.06-1.96(m,1H)。HRMS(ESI+):计算值C26H27N4O5(M+H)+,475.1976；实测值475.1972。
实施例14：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-乙烯基-2H-色烯-3-甲酰胺的合成(化合物A-14)
步骤1：将5-溴-2-羟基苯甲醛(36，1.50g，7.50mmol)溶于20mL的N,N-二甲基甲酰胺中，依次加入氯化锂(0.95g，22.51mmol)、四(三苯基膦)钯(0.87g，0.75mmol)和三丁基乙烯基锡(2.85g，9.00mmol)，加毕，升温至95℃搅拌4小时。经TLC监测反应完全，将反应液倒入100mL冰水中，析出棕褐色固体，静置使其完全析出，抽滤，将滤饼使用硅胶色谱柱(二氯甲烷：甲醇＝80：1，v/v)纯化，得到黄色固体37：2-羟基-5-乙烯基苯甲醛(0.68g，收率60.9％)。HRMS(ESI+):计算值C9H9O2(M+H)+,149.0597；实测值149.0588。
步骤2-步骤4：采用原料2-羟基-5-乙烯基苯甲醛参照中间体-2和化合物A-1的合成方法制备，得到白色固体A-14(0.07g，收率50.5％，纯度98.49％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.95(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.46(d,J＝7.3Hz,1H),7.33(s,1H),7.25(s,1H),6.96(t,J＝7.4Hz,1H),6.85(d,J＝8.1Hz,1H),6.61-6.45(m,1H),5.70(d,J＝6.8Hz,1H),5.50-5.26(m,1H),5.11(dd,J＝13.3,5.1Hz,1H),4.94(s,2H),4.56-4.24(m,4H),3.01-2.81(m,1H),2.60(d,J＝17.5Hz,1H),2.49-2.28(m,1H),2.08-1.93(m,1H)。HRMS(ESI+):计算值C26H24N3O5(M+H)+,458.1711；实测值458.1702。
实施例15：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-乙炔基-2H-色烯-3-甲酰胺的合成(化合物A-15)
6-(乙炔基)-2H-色烯-3-羧酸(40)的合成：采用原料5-乙炔基水杨醛参照中间体-2的合成方法制备，两步收率共计44.5％。HRMS(ESI-):计算值C12H7O3(M-H)-,199.0401；实测值199.0411。
化合物A-15的合成：采用原料6-(乙炔基)-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-15(0.08g，收率54.7％，纯度98.90％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.92(t,J＝6.0Hz,1H),7.72(d,J＝7.8Hz,1H),7.53(s,1H),7.46(d,J＝7.3Hz,1H),7.33(s,1H),7.25(s,1H),6.99-6.92(m,1H),6.85(d,J＝8.1Hz,1H),5.11(dd,J＝13.3,5.1Hz,1H),4.94(s,2H),4.56-4.24(m,4H),4.15(s,1H),2.99-2.84(m,1H),2.60(d,J＝17.5Hz,1H),2.47-2.25(m,1H),2.08-1.95(m,1H)。HRMS(ESI+):计算值C26H22N3O5(M+H)+,456.1554；实测值456.1547。
实施例16：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-叔丁基-2H-色烯-3-甲酰胺的合成(化合物A-16)
6-(叔丁基)-2H-色烯-3-羧酸(41)的合成：采用原料5-叔丁基水杨醛参照中间体-2的合成方法制备，两步收率共计47.6％。HRMS(ESI-):计算值C14H15O3(M-H)-,231.1027；实测值231.1023。
化合物A-16的合成：采用原料6-(叔丁基)-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-16(0.06g，收率38.4％，纯度97.69％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.03(s,1H),8.90(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.45(d,J＝7.5Hz,1H),7.36(s,1H),7.30-7.19(m,2H),6.79(d,J＝8.3Hz,1H),5.12(dd,J＝13.3,5.0Hz,1H),4.90(s,2H),4.56-4.24(m,4H),3.00-2.84(m,1H),2.60(d,J＝16.9Hz,1H),2.48-2.31(m,1H),2.06-1.93(m,1H),1.25(s,9H)。HRMS(ESI+):计算值C28H30N3O5(M+H)+,487.2107；实测值487.2103。
实施例17：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-苯基-2H-色烯-3-甲酰胺的合成(化合物A-17)
步骤1：将5-溴-2-羟基苯甲醛(36，1.80g，9.00mmol)溶于30mL的水中，依次加入苯硼酸(1.65g，13.50mmol)、碳酸钾(2.49g，18.00mmol)和四(三苯基膦)钯(0.02g，0.02mmol)，加毕。反应液经氮气保护后，升温至80℃搅拌6小时。经TLC监测反应完全，抽滤，滤饼水洗后真空干燥得到灰白色固体42：4-羟基-[1,1'-联苯]-3-甲醛(1.60g，收率89.8％)。HRMS(ESI+):计算值C13H11O2(M+H)+,199.0754；实测值199.0759。
步骤2-步骤4：采用原料4-羟基-[1,1'-联苯]-3-甲醛参照中间体-2和化合物A-1的合成方法制备，得到白色固体A-17(0.10g，收率60.8％，纯度98.56％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.90(t,J＝6.0Hz,1H),7.76-7.68(m,3H),7.55-7.40(m,5H),7.33(s,1H),7.29-7.18(m,2H),6.96-6.89(m,1H),5.11(dd,J＝13.3,5.1Hz,1H),4.85(s,2H),4.55-4.27(m,4H),2.99-2.81(m,1H),2.60(d,J＝17.4Hz,1H),2.49-2.28(m,1H),2.10-1.95(m,1H)。HRMS(ESI+):计算值C30H26N3O5(M+H)+,508.1867；实测值508.1860。
实施例18：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-8-三氟甲基-2H-色烯-3-甲酰胺的合成(化合物A-18)
8-三氟甲基-2H-色烯-3-羧酸(45)的合成：采用原料2-羟基-3-三氟甲基苯甲醛参照中间体-2的合成方法制备，两步收率共计50.6％。HRMS(ESI-):计算值C11H6F3O3(M-H)-,243.0275；实测值243.0265。
化合物A-18的合成：采用原料8-三氟甲基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-18(0.07g，收率43.5％，纯度97.20％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.04(s,1H),9.02(t,J＝6.0Hz,1H),7.71(d,J＝7.8Hz,1H),7.61-7.51(m,3H),7.46(d,J＝7.9Hz,1H),7.39(s,1H),7.12(t,J＝7.7Hz,1H),5.17-5.10(m,1H),5.09(s,2H),4.56-4.24(m,4H),3.00-2.84(m,1H),2.60(d,J＝16.9Hz,1H),2.48-2.30(m,1H),2.05-1.94(m,1H)。HRMS(ESI+):计算值C25H21F3N3O5(M+H)+,500.1428；实测值500.1422。
实施例19：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-8-三氟甲氧基-2H-色烯-3-甲酰胺的合成(化合物A-19)
8-三氟甲氧基-2H-色烯-3-羧酸(46)的合成：采用原料2-羟基-3-三氟甲氧基苯甲醛参照中间体-2的合成方法制备，两步收率共计48.2％。HRMS(ESI-):计算值C11H6F3O4(M-H)-,259.0224；实测值259.0220。
化合物A-19的合成：采用原料8-三氟甲氧基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-19(0.06g，收率35.9％，纯度98.56％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.03(s,1H),9.02(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.46(d,J＝7.9Hz,1H),7.39(s,1H),7.36-7.25(m,2H),7.04(t,J＝7.9Hz,1H),5.12(dd,J＝13.2,5.0Hz,1H),5.04(s,2H),4.58-4.24(m,4H),3.01-2.84(m,1H),2.60(d,J＝16.9Hz,1H),2.47-2.29(m,1H),2.06-1.94(m,1H)。HRMS(ESI+):计算值C25H21F3N3O6(M+H)+,515.1406；实测值515.1402。
实施例20：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-8-甲氧基-2H-色烯-3-甲酰胺的合成(化合物A-20)
8-甲氧基-2H-色烯-3-羧酸(47)的合成：采用原料2-羟基-3-甲氧基苯甲醛参照中间体-2的合成方法制备，两步收率共计56.5％。HRMS(ESI-):计算值C11H9O4(M-H)-,205.0506；实测值205.0500。
化合物A-20的合成：采用原料8-甲氧基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-20(0.07g，收率50.0％，纯度98.56％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.03(s,1H),8.94(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.52(s,1H),7.45(d,J＝7.8Hz,1H),7.32(s,1H),6.99(d,J＝6.5Hz,1H),6.91(t,J＝7.7Hz,1H),6.83(d,J＝7.5Hz,1H),5.12(dd,J＝13.4,5.0Hz,1H),4.91(s,2H),4.57-4.23(m,4H),3.76(s,3H),3.02-2.82(m,1H),2.60(d,J＝16.8Hz,1H),2.44-2.29(m,1H),2.06-1.88(m,1H)。HRMS(ESI+):计算值C25H24N3O6(M+H)+,462.5943；实测值462.5938。
实施例21：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-8-甲基-2H-色烯-3-甲酰胺的合成(化合物A-21)
8-甲基-2H-色烯-3-羧酸(48)的合成：采用原料2-羟基-3-甲基苯甲醛参照中间体-2的合成方法制备，两步收率共计60.2％。HRMS(ESI-):计算值C11H9O3(M-H)-,189.0557；实测值189.0545。
化合物A-21的合成：采用原料8-甲基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-21(0.07g，收率48.6％，纯度98.26％)。1H NMR(300MHz,DMSO-d6)δ(ppm)1.00(s,1H),8.89(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.49(s,1H),7.44(d,J＝7.8Hz,1H),7.29(s,1H),7.04(d,J＝8.1Hz,2H),6.75(d,J＝7.9Hz,1H),5.11(dd,J＝13.2,5.0Hz,1H),4.90(s,2H),4.56-4.24(m,4H),2.99-2.83(m,1H),2.60(d,J＝17.0Hz,1H),2.48-2.31(m,1H),2.19(s,3H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C25H24N3O5(M+H)+,446.1711；实测值446.1707。
实施例22：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-5-甲基-2H-色烯-3-甲酰胺的合成(化合物A-22)
5-甲基-2H-色烯-3-羧酸(49)的合成：采用原料2-羟基-6-甲基苯甲醛参照中间体-2的合成方法制备，两步收率共计60.6％。HRMS(ESI-):计算值C11H9O3(M-H)-,189.0557；实测值189.0546。
化合物A-22的合成：采用原料5-甲基-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-22(0.09g，收率62.5％，纯度97.88％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.89(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.50(s,1H),7.35(d,J＝7.8Hz,1H),7.16(s,1H),7.04(d,J＝8.1Hz,2H),6.75(d,J＝7.9Hz,1H),5.13(dd,J＝13.2,5.0Hz,1H),4.92(s,2H),4.56-4.24(m,4H),2.99-2.84(m,1H),2.59(d,J＝17.0Hz,1H),2.47-2.30(m,1H),2.12(s,3H),2.05-1.90(m,1H)。HRMS(ESI+):计算值C25H24N3O5(M+H)+,446.1711；实测值446.1709。
实施例23：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-(乙酰氨基)-2H-色烯-3-甲酰胺的合成(化合物A-23)
将N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氨基-2H-色烯-3-甲酰胺(化合物12，0.10g，0.22mmol)溶解于5mL的乙腈中，依次加入三乙胺(0.03g，0.34mmol)、乙酰氯(0.02g，0.25mmol)，加毕，室温搅拌1小时。经TLC监测反应完全，反应液真空浓缩，将残余物使用硅胶色谱柱(二氯甲烷：甲醇＝50：1，v/v)纯化，得到白色固体A-23(0.04g，收率36.7％，纯度98.20％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.02(s,1H),10.22(s,1H),8.98(t,J＝6.0Hz,1H),7.67(d,J＝7.8Hz,1H),7.56(s,1H),7.45(d,J＝7.3Hz,1H),7.31(d,J＝8.1,1H),7.10-6.89(m,3H),5.11(dd,J＝13.3,5.1Hz,1H),4.88(s,2H),4.48-4.24(m,4H),2.97-2.80(m,1H),2.58(d,J＝17.5Hz,1H),2.48-2.32(m,1H),2.13(s,3H),2.07-1.96(m,1H)。HRMS(ESI+):计算值C26H25N4O6(M+H)+,489.1769；实测值489.1760。
实施例24：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-(丙烯酰胺)-2H-色烯-3-甲酰胺的合成(化合物A-24)
采用原料丙烯酰氯参照化合物A-15的合成方法制备，得到白色固体A-24(0.05g，收率44.6％，纯度95.96％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),9.54(s,1H),8.90(t,J＝6.0Hz,1H),7.67(d,J＝7.8Hz,1H),7.56(s,1H),7.45(d,J＝7.3Hz,1H),7.33(s,1H),7.30-7.22(m,2H),6.99(d,J＝8.1Hz,1H),6.50-6.44(m,1H),6.08-6.00(m,2H),5.11(dd,J＝13.3,5.1Hz,1H),4.81(s,2H),4.48-4.24(m,4H),2.97-2.80(m,1H),2.58(d,J＝17.5Hz,1H),2.48-2.32(m,1H),2.07-1.96(m,1H)。
HRMS(ESI+):计算值C27H25N4O6(M+H)+,501.1769；实测值501.1757。
实施例25：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-(2-羟基乙酰胺)-2H-色烯-3-甲酰胺的合成(化合物A-25)
将N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氨基-2H-色烯-3-甲酰胺(化合物12，0.07g，0.16mmol)溶解于5mL的N,N-二甲基甲酰胺中，依次加入羟基乙酸(0.01g，0.19mmol)、HATU(0.09g，0.24mmol)，加毕，室温搅拌1小时。加入DIPEA(0.06g，0.47mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入30mL冰水中，使用乙酸乙酯萃取，合并有机相并依次使用水和饱和食盐水洗涤，无水硫酸钠干燥后真空浓缩，将残余物使用硅胶色谱柱(二氯甲烷：甲醇＝50：1,v/v)纯化，得到淡黄色固体A-25(0.03g，收率43.0％，纯度97.48％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),9.54(s,1H),8.90(t,J＝6.0Hz,1H),7.67(d,J＝7.8Hz,1H),7.57(s,1H),7.45(d,J＝7.3Hz,1H),7.33(s,1H),7.28(d,J＝8.0Hz,1H),7.00(d,J＝8.1Hz,1H),6.88(d,J＝7.9Hz,1H),5.15(s,1H),5.07(dd,J＝13.3,5.1Hz,1H),4.79(s,2H),4.55-4.29(m,6H),2.97-2.80(m,1H),2.61(d,J＝17.5Hz,1H),2.47-2.30(m,1H),2.07-1.96(m,1H)。HRMS(ESI+):计算值C26H25N4O7(M+H)+,505.1718；实测值505.1712。
实施例26：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-(环丙烷甲酰胺)-2H-色烯-3-甲酰胺的合成(化合物A-26)
采用原料环丙基甲酰氯参照化合物A-15的合成方法制备，得到白色固体A-26(0.05g，收率43.5％，纯度96.40％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),9.99(s,1H),8.99(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.59(s,1H),7.47(d,J＝7.3Hz,1H),7.35(s,1H),7.25(d,J＝8.0Hz,1H),6.99-6.88(m,2H),5.02(dd,J＝13.3,5.1Hz,1H),4.80(s,2H),4.55-4.30(m,4H),2.97-2.81(m,1H),2.59(d,J＝17.6Hz,1H),2.47-2.31(m,1H),2.07-1.99(m,1H),1.49-1.45(m,1H),0.80-0.61(m,4H)。HRMS(ESI+):计算值C28H27N4O6(M+H)+,515.1925；实测值515.1918。
实施例27：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-(环丁烷甲酰胺)-2H-色烯-3-甲酰胺的合成(化合物A-27)
采用原料环丁基甲酰氯参照化合物A-15的合成方法制备，得到白色固体A-27(0.06g，收率50.8％，纯度97.68％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),9.89(s,1H),8.98(t,J＝6.0Hz,1H),7.68(d,J＝7.8Hz,1H),7.55(s,1H),7.44(d,J＝7.3Hz,1H),7.35(s,1H),7.25(d,J＝8.0Hz,1H),7.01(d,J＝8.2Hz,1H),6.90(d,J＝7.9Hz,1H),5.04(dd,J＝13.3,5.1Hz,1H),4.80(s,2H),4.52-4.29(m,4H),2.97-2.82(m,1H),2.59(d,J＝17.6Hz,1H),2.47-2.31(m,1H),2.07-1.85(m,4H),1.78-1.53(m,4H)。HRMS(ESI+):计算值C29H29N4O6(M+H)+,529.2082；实测值529.2077。
实施例28：N-(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氯-7-甲基-2H-色烯-3-甲酰胺的合成(化合物A-28)
6-氯-7-甲基-2H-色烯-3-羧酸(50)的合成：采用原料5-氯-4-甲基水杨醛参照中间体-2的合成方法制备，两步收率共计54.6％。HRMS(ESI-):计算值C11H8ClO3(M-H)-,223.0167；实测值223.0158。
化合物A-28的合成：采用原料6-氯-7-甲基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-28(0.08g，收率52.2％，纯度97.43％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.89(t,J＝6.0Hz,1H),7.69(d,J＝7.8Hz,1H),7.52(s,1H),7.44(d,J＝7.9Hz,1H),7.28(d,J＝5.9Hz,2H),6.88(s,1H),5.11(dd,J＝13.2,5.0Hz,1H),4.94(s,2H),4.55-4.25(m,4H),3.03-2.85(m,1H),2.60(d,J＝17.8Hz,1H),2.46-2.33(m,1H),2.27(s,3H),2.07-1.96(m,1H)。HRMS(ESI+):计算值C25H23ClN3O5(M+H)+,480.1321；实测值480.1309。
实施例29：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-7-氯-6-甲基-2H-色烯-3-甲酰胺的合成(化合物A-29)
步骤1：将3-氯-4-甲基-苯酚(51，1.00g，7.04mmol)溶于20mL的乙腈中，依次加入甲醛(1.11g，36.99mmol)、三乙胺(3.74g，36.99mmol)和氯化镁(2.68g，28.16mmol)，加毕，升温至80℃搅拌15小时。经TLC监测反应完全，添加50mL盐酸(1M)淬灭，使用乙酸乙酯萃取，合并有机相并使用饱和食盐水洗涤，无水硫酸钠干燥后真空浓缩得到棕色油状物52：4-氯-2-羟基-5-甲基苯甲醛(1.02g，收率84.9％)，该产物无需进一步纯化可直接用于下一步。HRMS(ESI+):计算值C8H8ClO2(M+H)+,171.0207；实测值171.0217。
步骤2-步骤4：采用原料4-氯-2-羟基-5-甲基苯甲醛参照中间体-2和化合物A-1的合成方法制备，得到白色固体A-29(0.03g，收率19.4％，纯度98.23％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.89(t,J＝6.0Hz,1H),7.69(d,J＝7.8Hz,1H),7.52(s,1H),7.44(d,J＝7.9Hz,1H),7.23(d,J＝5.9Hz,2H),6.94(s,1H),5.11(dd,J＝13.2,5.0Hz,1H),4.94(s,2H),4.53-4.25(m,4H),3.00-2.85(m,1H),2.60(d,J＝17.8Hz,1H),2.47-2.35(m,1H),2.22(s,3H),2.07-1.96(m,1H)。HRMS(ESI+):计算值C25H23ClN3O5(M+H)+,480.1321；实测值480.1318。
实施例30：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-7-氟-6-甲基-2H-色烯-3-甲酰胺的合成(化合物A-30)
7-氟-6-甲基-2H-色烯-3-羧酸(55)的合成：采用原料4-氟-5-甲基水杨醛参照中间体-2的合成方法制备，两步收率共计51.2％。HRMS(ESI-):计算值C11H8FO3(M-H)-,207.0463；实测值207.0458。
化合物A-30的合成：采用原料7-氟-6-甲基-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-30(0.07g，收率47.3％，纯度98.32％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),8.92(t,J＝6.0Hz,1H),7.69(d,J＝7.8Hz,1H),7.50(s,1H),7.40(d,J＝7.5Hz,1H),7.31(d,J＝5.9Hz,2H),6.96(s,1H),5.11(dd,J＝13.2,5.0Hz,1H),4.94(s,2H),4.55-4.25(m,4H),3.03-2.85(m,1H),2.59(d,J＝17.5Hz,1H),2.48-2.35(m,1H),2.16(s,3H),2.08-1.95(m,1H)。HRMS(ESI+):计算值C25H23FN3O5(M+H)+,464.1616；实测值464.1612。
实施例31：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氟-8-甲基-2H-色烯-3-甲酰胺的合成(化合物A-31)
6-氟-8-甲基-2H-色烯-3-羧酸(56)的合成：采用原料5-氟-3-甲基水杨醛参照中间体-2的合成方法制备，两步收率共计57.3％。HRMS(ESI-):计算值C11H8FO3(M-H)-,207.0463；实测值207.0465。
化合物A-31的合成：采用原料5-氟-3-甲基-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-31(0.08g，收率54.0％，纯度99.82％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.89(t,J＝6.0Hz,1H),7.69(d,J＝7.8Hz,1H),7.55(s,1H),7.46(d,J＝7.9Hz,1H),7.32(d,J＝5.9Hz,2H),6.99(s,1H),5.12(dd,J＝13.2,5.0Hz,1H),4.94(s,2H),4.55-4.25(m,4H),3.00-2.85(m,1H),2.60(d,J＝17.8Hz,1H),2.45-2.30(m,1H),2.20(s,3H),2.07-1.95(m,1H)。HRMS(ESI+):计算值C25H23FN3O5(M+H)+,464.1616；实测值464.1604。
实施例32：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-7,8-二氟-2H-色烯-3-甲酰胺的合成(化合物A-32)
7,8-二氟-2H-色烯-3-羧酸(57)的合成：采用原料3,4-二氟水杨醛参照中间体-2的合成方法制备，两步收率共计52.4％。HRMS(ESI-):计算值C10H5F2O3(M-H)-,211.0212；实测值211.0215。
化合物A-32的合成：采用原料7,8-二氟-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-32(0.06g，收率40.2％，纯度98.32％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.95(s,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.40(d,J＝8.1Hz,1H),7.29(d,J＝8.9Hz,2H),7.15(d,J＝8.4Hz,1H),5.11(dd,J＝13.2,5.1Hz,1H),5.00(s,2H),4.56-4.24(m,4H),3.04-2.79(m,1H),2.60(d,J＝16.9Hz,1H),2.47-2.33(m,1H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C24H20F2N3O5(M+H)+,468.1366；实测值468.1361。
实施例33：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6,8-二氟-2H-色烯-3-甲酰胺的合成(化合物A-33)
6,8-二氟-2H-色烯-3-羧酸(58)的合成：采用原料3,5-二氟水杨醛参照中间体-2的合成方法制备，两步收率共计52.4％。HRMS(ESI-):计算值C10H5F2O3(M-H)-,211.0212；实测值211.0220。
化合物A-33的合成：采用原料6,8-二氟-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-33(0.07g，收率46.9％，纯度97.14％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),9.02(s,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.45(d,J＝8.1Hz,1H),7.30(d,J＝8.9Hz,2H),7.05(d,J＝8.4Hz,1H),5.11(dd,J＝13.2,5.1Hz,1H),5.01(s,2H),4.56-4.23(m,4H),3.04-2.79(m,1H),2.60(d,J＝16.9Hz,1H),2.44-2.33(m,1H),2.07-1.90(m,1H)。HRMS(ESI+):计算值C24H20F2N3O5(M+H)+,468.1366；实测值468.1362。
实施例34：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-5,7-二氟-2H-色烯-3-甲酰胺的合成(化合物A-34)
5,7-二氟-2H-色烯-3-羧酸(59)的合成：采用原料2,4-二氟水杨醛参照中间体-2的合成方法制备，两步收率共计54.8％。HRMS(ESI-):计算值C10H5F2O3(M-H)-,211.0212；实测值211.0217。
化合物A-34的合成：采用原料5,7-二氟-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-34(0.08g，收率53.6％，纯度97.92％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.96(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.55(s,1H),7.40-7.31(m,2H),7.22(s,1H),6.98(dd,J＝11.5,6.8Hz,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.97(s,2H),4.57-4.26(m,4H),3.00-2.83(m,1H),2.59(d,J＝17.2Hz,1H),2.47-2.32(m,1H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C24H20F2N3O5(M+H)+,468.1366；实测值468.1357。
实施例35：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6,7-二氟-2H-色烯-3-甲酰胺的合成(化合物A-35)
6,7-二氟-2H-色烯-3-羧酸(60)的合成：采用原料4,5-二氟水杨醛参照中间体-2的合成方法制备，两步收率共计54.1％。HRMS(ESI-):计算值C10H5F2O3(M-H)-,211.0212；实测值211.0223。
化合物A-35的合成：采用原料6,7-二氟-2H-色烯-3-羧酸通过化合物A-1的合成方法制备，得白色固体A-35(0.05g，收率33.5％，纯度97.09％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.93(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.52(s,1H),7.47-7.35(m,2H),7.27(s,1H),7.04(dd,J＝11.7,6.9Hz,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.97(s,2H),4.57-4.24(m,4H),3.00-2.81(m,1H),2.60(d,J＝17.2Hz,1H),2.47-2.30(m,1H),2.07-1.91(m,1H)。HRMS(ESI+):计算值C24H20F2N3O5(M+H)+,468.1366；实测值468.1359。
实施例36：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-5,8-二氟-2H-色烯-3-甲酰胺的合成(化合物A-36)
5,8-二氟-2H-色烯-3-羧酸(61)的合成：采用原料3,6-二氟水杨醛参照中间体-2的合成方法制备，两步收率共计49.8％。HRMS(ESI-):计算值C10H5F2O3(M-H)-,211.0212；实测值211.0219。
化合物A-36的合成：采用原料5,8-二氟-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-36(0.07g，收率46.9％，纯度98.62％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.93(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.53(s,1H),7.45(d,J＝8.1Hz,1H),7.32(s,1H),7.20(s,1H),7.04(dd,J＝11.7,6.9Hz,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.98(s,2H),4.55-4.24(m,4H),2.99-2.80(m,1H),2.60(d,J＝17.2Hz,1H),2.47-2.31(m,1H),2.07-1.91(m,1H)。HRMS(ESI+):计算值C24H20F2N3O5(M+H)+,468.1366；实测值468.1360。
实施例37：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6,7-二甲基-2H-色烯-3-甲酰胺的合成(化合物A-37)
6,8-二甲基-2H-色烯-3-羧酸(62)的合成：采用原料3,5-二甲基水杨醛参照中间体-2的合成方法制备，两步收率共计54.1％。HRMS(ESI-):计算值C12H11O3(M-H)-,203.0714；实测值203.0716。
化合物A-37的合成：采用原料6,8-二甲基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-37(0.06g，收率40.8％，纯度97.09％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.89(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.50(s,1H),7.42(d,J＝7.7Hz,1H),7.30(s,1H),7.02(d,J＝8.1Hz,2H),5.11(dd,J＝13.2,5.0Hz,1H),4.91(s,2H),4.56-4.23(m,4H),2.99-2.83(m,1H),2.60(d,J＝17.5Hz,1H),2.48-2.30(m,1H),2.24(s,3H),2.12(s,3H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C26H26N3O5(M+H)+,460.1867；实测值460.1860。
实施例38：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-5,8-二甲基-2H-色烯-3-甲酰胺的合成(化合物A-38)
5,8-二甲基-2H-色烯-3-羧酸(63)的合成：采用原料3,6-二甲基水杨醛参照中间体-2的合成方法制备，两步收率共计52.6％。HRMS(ESI-):计算值C12H11O3(M-H)-,203.0714；实测值203.0720。
化合物A-38的合成：采用原料5,8-二甲基-2H-色烯-3-羧酸参照化合物A-1的合成方法制备，得白色固体A-38(0.07g，收率47.6％，纯度96.34％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.98(s,1H),8.99(t,J＝6.0Hz,1H),7.75(d,J＝7.6Hz,1H),7.50(s,1H),7.42(d,J＝7.7Hz,1H),7.30(s,1H),7.00(d,J＝8.5Hz,2H),5.11(dd,J＝13.2,5.0Hz,1H),4.91(s,2H),4.55-4.24(m,4H),2.99-2.83(m,1H),2.60(d,J＝17.6Hz,1H),2.47(s,3H),2.45-2.33(m,1H),2.15(s,3H)2.05-1.93(m,1H)。HRMS(ESI+):计算值C26H26N3O5(M+H)+,460.1867；实测值460.1853。
实施例39：N-((2-(2,6-二氧代哌啶-3-基)-3-氧代异吲哚啉-4-基)甲基)-6-(三氟甲氧基)-2H-色烯-3-甲酰胺的合成(化合物A-39)
步骤1-步骤6：采用原料2-溴-6-甲基苯甲酸甲酯(64)参照中间体-1和化合物A-1的合成方法制备，得到白色固体A-39(0.10g，收率57.1％，纯度98.16％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),8.96(t,J＝6.0Hz,1H),7.78-7.70(m,2H),7.45(d,J＝7.8Hz,1H),7.33(s,1H),6.96-6.79(m,3H),5.12(dd,J＝13.3,5.1Hz,1H),5.00(s,2H),4.56-4.25(m,4H),3.00-2.81(m,1H),2.62(d,J＝17.6Hz,1H),2.48-2.30(m,1H),2.07-1.94(m,1H)。HRMS(ESI+):计算值C24H21F3N3O6(M+H)+,516.1377；实测值516.1368。
实施例40：N-((2-(2,6-二氧代哌啶-3-基)-3-氧代异吲哚啉-5-基)甲基)-6-(三氟甲氧基)-2H-色烯-3-甲酰胺的合成(化合物A-40)
步骤1-步骤6：采用原料5-溴-2-甲基苯甲酸甲酯(70)参照中间体-1和化合物A-1的合成方法制备，得到白色固体A-40(0.10g，收率62.7％，纯度99.20％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),8.96(t,J＝6.0Hz,1H),7.84(s,1H),7.75(d,J＝7.6Hz,1H),7.50-7.38(m,2H),6.90-6.75(m,3H),5.12(dd,J＝13.3,5.1Hz,1H),5.00(s,2H),4.56-4.29(m,4H),3.00-2.81(m,1H),2.60(d,J＝17.1Hz,1H),2.49-2.35(m,1H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C24H21F3N3O6(M+H)+,516.1377；found,516.1366。
实施例41：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)甲基)-6-(三氟甲氧基)-2H-色烯-3-甲酰胺的合成(化合物A-41)
步骤1-步骤6：采用原料3-溴-2-甲基苯甲酸甲酯(76)参照中间体-1和化合物A-1的合成方法制备，得到白色固体A-41(0.09g，收率54.2％，纯度97.59％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.99(t,J＝6.0Hz,1H),7.85(s,1H),7.75(d,J＝7.6Hz,1H),7.55-7.39(m,2H),6.92-6.74(m,3H),5.12(dd,J＝13.3,5.1Hz,1H),4.95(s,2H),4.55-4.27(m,4H),3.00-2.82(m,1H),2.62(d,J＝17.6Hz,1H),2.49-2.34(m,1H),2.07-1.94(m,1H)。HRMS(ESI+):计算值C24H21F3N3O6(M+H)+,516.1377；found,516.1374。
实施例42：N-((2-(2，6-二氧代哌啶-3-基)-4-氟-1-氧代异吲哚啉-5-基)甲基)-6-(三氟甲氧基)-2H-色烯-3-甲酰胺的合成(化合物A-42)
化合物A-42的合成：采用原料中间体-3和6-三氟甲氧基-2H-色烯-3-羧酸(24)参照化合物A-1的合成方法制备，得白色固体A-42(0.09g，收率56.3％，纯度97.98％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.02(s,1H),8.93(t,J＝5.4Hz,1H),7.70-7.51(m,2H),7.33(d,J＝9.1Hz,2H),7.25(d,J＝8.7Hz,1H),6.97(d,J＝8.8Hz,1H),5.14(dd,J＝13.2,4.8Hz,1H),4.98(s,2H),4.60-4.36(m,4H),2.97-2.86(m,1H),2.63(d,J＝17.2Hz,1H),2.46-2.38(m,1H),2.03-1.99(m,1H)ppm.HRMS(ESI+)：计算值C25H20F4N3O6(M+H)+，534.1283；实测值534.1291。
实施例43：N-((2-(2，6-二氧代哌啶-3-基)-4-氟-1-氧代异吲哚啉-5-基)甲基)-8-(三氟甲氧基)-2H-色烯-3-甲酰胺的合成(化合物A-43)
化合物A-43的合成：采用原料中间体-3和8-三氟甲氧基-2H-色烯-3-羧酸(46)参照化合物A-1的合成方法制备，得白色固体A-43(0.09g，收率56.5％，95.82纯度％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.03(s,1H),8.97(t,J＝6.0Hz,1H),7.60-7.52(m,2H),7.39(s,1H),7.30(t,J＝9.9Hz,2H),7.03(t,J＝7.9Hz,1H),5.15(dd,J＝13.0,4.8Hz,1H),5.02(s,2H),4.60-4.36(m,4H),2.97-2.86(m,1H),2.63(d,J＝17.7Hz,1H),2.46-2.36(m,1H),2.03-1.90(m,1H)ppm.HRMS(ESI+)：计算值C25H20F4N3O6(M+H)+，534.1283；实测值534.1263。
实施例44：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)苯并二氢吡喃-3-甲酰胺的合成(化合物B-1)
采用原料苯并二氢吡喃-3-羧酸参照化合物A-1的合成方法制备，得白色固体B-1(0.08g，收率57.9％，纯度98.03％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.03(s,1H),8.78(t,J＝6.0Hz,1H),7.71(d,J＝7.8Hz,1H),7.48(s,1H),7.43(d,J＝7.9Hz,1H),7.18-7.05(m,2H),6.86(t,J＝7.3Hz,1H),6.79(d,J＝8.1Hz,1H),5.14(dd,J＝13.3,5.0Hz,1H),4.50-4.28(m,5H),3.99(t,J＝9.7Hz,1H),3.04-2.86(m,4H),2.62(d,J＝17.7Hz,1H),2.47-2.33(m,1H),2.09-1.97(m,1H)。HRMS(ESI+):计算值C24H24N3O5(M+H)+,434.1711；实测值434.1702。
实施例45：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氟苯并二氢吡喃-3-甲酰胺的合成(化合物B-2)
6-氟苯并二氢吡喃-3-羧酸(82)的合成：采用原料5-氟水杨醛参照中间体-2的合成方法制备，三步收率共计51.2％。HRMS(ESI-):计算值C10H8FO3(M-H)-,195.0463；实测值195.0468。
化合物B-2的合成：采用原料6-氟苯并二氢吡喃-3-羧酸参照化合物A-1的合成方法制备，得白色固体B-2(0.06g，收率41.6％，纯度97.09％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.86(t,J＝6.0Hz,1H),7.75(d,J＝7.8Hz,1H),7.50(s,1H),7.40(d,J＝7.9Hz,1H),7.15-7.03(m,2H),6.79(d,J＝8.1Hz,1H),5.14(dd,J＝13.3,5.0Hz,1H),4.48-4.25(m,5H),3.99(t,J＝9.7Hz,1H),3.02-2.86(m,4H),2.61(d,J＝17.5Hz,1H),2.48-2.33(m,1H),2.07-1.95(m,1H)。HRMS(ESI+):计算值C24H23FN3O5(M+H)+,452.1616；实测值452.1613。
实施例46：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-氯苯并二氢吡喃-3-甲酰胺的合成(化合物B-3)
6-氯苯并二氢吡喃-3-羧酸(83)的合成：采用原料5-氯水杨醛参照中间体-2的合成方法制备，三步收率共计49.6％。HRMS(ESI-):计算值C10H8ClO3(M-H)-,211.0167；实测值211.0159。
化合物B-3的合成：采用原料6-氯苯并二氢吡喃-3-羧酸参照化合物A-1的合成方法制备，得白色固体B-3(0.04g，收率26.8％，纯度98.73％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.87(t,J＝6.0Hz,1H),7.70(d,J＝7.8Hz,1H),7.55(s,1H),7.40(d,J＝7.9Hz,1H),7.14-7.00(m,2H),6.79(d,J＝8.1Hz,1H),5.10(dd,J＝13.3,5.0Hz,1H),4.48-4.25(m,5H),4.01(t,J＝9.6Hz,1H),3.02-2.85(m,4H),2.60(d,J＝17.0Hz,1H),2.48-2.32(m,1H),2.07-1.95(m,1H)。HRMS(ESI+):计算值C24H23ClN3O5(M+H)+,468.1231；实测值468.1225。
实施例47：N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-6-(三氟甲氧基)苯并二氢吡喃-3-甲酰胺的合成(化合物B-4)
6-三氟甲氧基苯并二氢吡喃-3-羧酸(84)的合成：采用原料5-三氟甲氧基水杨醛参照中间体-2的合成方法制备，三步收率共计50.8％。HRMS(ESI-):计算值C11H8F3O4(M-H)-,261.0380；实测值261.0386。
化合物B-4的合成：采用原料6-三氟甲氧基苯并二氢吡喃-3-羧酸参照化合物A-1的合成方法制备，得白色固体B-4(0.06g，收率36.3％，纯度99.50％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.77(t,J＝5.9Hz,1H),7.68(d,J＝7.8Hz,1H),7.47(s,1H),7.40(d,J＝7.9Hz,1H),7.17(s,1H),7.09(d,J＝8.9Hz,1H),6.87(d,J＝8.9Hz,1H),5.12(dd,J＝13.3,5.0Hz,1H),4.48-4.26(m,5H),4.04(t,J＝9.7Hz,1H),3.09-2.85(m,4H),2.60(d,J＝17.4Hz,1H),2.48-2.32(m,1H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C25H23F3N3O6(M+H)+,518.1534；实测值518.1540。
实施例48：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-1,2,3,4-四氢喹啉-3-甲酰胺的合成(化合物C-1)
采用原料1,2,3,4-四氢喹啉-3-羧酸参照化合物A-1的合成方法制备，得白色固体C-1(0.09g，收率63.3％，纯度98.67％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),8.64(t,J＝5.9Hz,1H),7.68(d,J＝7.8Hz,1H),7.45(s,1H),7.39(d,J＝7.8Hz,1H),6.87(t,J＝8.1Hz,2H),6.49-6.38(m,2H),5.80(s,1H),5.12(dd,J＝13.3,5.0Hz,1H),4.49-4.26(m,4H),3.20-2.71(m,5H),2.68-2.55(m,2H),2.48-2.32(m,1H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C24H25N4O4(M+H)+,433.1871；实测值433.1866。
实施例49：N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-1,2,3,4-四氢萘-2-甲酰胺的合成(化合物D-1)
采用原料1,2,3,4-四氢萘-2-羧酸参照化合物A-1的合成方法制备，得白色固体D-1(0.12g，86.9％，纯度97.13％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.54(t,J＝6.0Hz,1H),7.69(d,J＝7.8Hz,1H),7.48(s,1H),7.41(d,J＝7.2Hz,1H),7.08(s,4H),5.11(dd,J＝13.3,5.1Hz,1H),4.52-4.24(m,4H),2.96-2.83(m,3H),2.83-2.72(m,2H),2.66-2.54(m,2H),2.47-2.32(m,1H),2.07-1.96(m,2H),1.82-1.64(m,1H)。HRMS(ESI+):计算值C25H26N3O4(M+H)+,432.1918；实测值432.1910。
生物测试例1：对KG-1细胞中的GSPT1降解作用的研究
KG-1细胞(ATCC)在IMDM(含20％的FBS)的培养基中生长，离心计数后调整细胞浓度以106个细胞/孔铺板于6孔板，每孔1350微升。加入150微升DMSO和本发明待测化合物，化合物浓度为200nM，置于5％CO2，37℃培养箱培养4小时。将细胞离心弃去培养液，加入PBS清洗后弃去。用含有蛋白酶抑制剂混合液(100×，弗德生物)、蛋白磷酸酶抑制剂混合液(100×，弗德生物)、超级核酸酶(碧云天)和高强度RIPA裂解液(赛默飞)制备全细胞裂解物，置于冰上30分钟。离心弃去细胞碎片沉淀，收集上清全细胞溶解产物并将其转移至新的离心管中。进行BCA蛋白测定后使用5X loading buffer(赛默飞)配制样品。将样品在4-20％的预制胶(SDS-PAGE凝胶)中进行电泳分离蛋白，再转至PVDF膜后用5％的NFDM/TBST进行室温封闭1小时，一抗4℃孵育过夜，次日二抗室温2小时孵育。使用MINICHEMITM成像系统检测信号。
一抗：
Anti-GSPT1：Abcam ab234433
Anti-beta Actin：弗德生物FD0060
二抗：
anti-rabbit peroxidase-linked secondary antibody：弗德生物FDG007
化合物对GSPT1蛋白降解效果如表1所示，其中A表示GSPT1蛋白降解百分数不小于80％，B表示降解百分数小于80％但不小于50％，C表示降解百分数小于50％但不小于25％，D表示降解百分数小于25％。
表1化合物对GSPT1蛋白的降解
从表中可以看出，与临床药物CC-90009相比，本发明的苯并六元杂环化合物如A、B、C系列对GSPT1蛋白的降解效果显著提高，而苯并六元脂肪环丧失对GSPT1蛋白的降解作用。并且从图1，2看出，本发明化合物A-10对KG-1、MV4-11细胞中的GSPT1降解作用呈现浓度依赖与时间依赖，效果较CC-90009更加显著。
生物测试例2：抗KG-1细胞增殖活性
将对数生长期的KG-1细胞用培养基(RPMI+10％的FBS)稀释，以5000个细胞/孔铺板于96孔板铺板，置于5％CO2，37℃培养箱培养24小时。将本发明待测化合物用DMSO配置成10mM母液，用培养基稀释设置9个浓度梯度，每个浓度3个复孔，加药后置于5％CO2，37℃培养箱培养72小时。使用CCK8法检测细胞增殖情况。用酶标法测定化合物对细胞的生长抑制IC50值。用GraphPad Prism 7软件计算IC50值。
化合物抗KG-1细胞增殖活性如表2所示，其中A表示IC50<1nM，B表示1≤IC50＜10nM，C表示10nM≤IC50＜100nM，D表示100nM≤IC50＜1000nM，E表示IC50≥1000nM。
表2化合物抗KG-1细胞增殖活性
从表中可以看出，与临床药物CC-90009相比，本发明的苯并六元杂环化合物如A、B、C系列对KG-1细胞的抗增殖抑制活性显著提高，而苯并六元脂肪环丧失对KG-1细胞的抗增殖抑制作用。
生物测试例3：本发明化合物对多种肿瘤细胞具有抗增殖活性
将对数生长期的U937、MOLM-13、MV4-11细胞(ATCC)分别用培养基稀释，以5000个细胞/孔铺板于96孔板铺板，置于5％CO2，37℃培养箱培养24小时。将化合物用DMSO配置成10mM母液，用培养基稀释设置9个浓度梯度，每个浓度3个复孔，加药后置于5％CO2，37℃培养箱培养72小时。使用CCK8法检测细胞增殖情况。用酶标法测定化合物对细胞的生长抑制IC50值。用GraphPad Prism 7软件计算IC50值。
化合物抗肿瘤细胞增殖活性如表3所示，其中A表示IC50<1nM，B表示1≤IC50＜10nM，C表示10nM≤IC50＜100nM，D表示100nM≤IC50＜1000nM，E表示IC50≥1000nM。
表3化合物抗肿瘤细胞增殖活性
从表中可以看出，与临床药物CC-90009相比，本发明的苯并六元杂环化合物如A、B、C系列对多种AML细胞的抗增殖抑制活性显著提高，而苯并六元脂肪环丧失对AML细胞的抗增殖抑制作用。
生物测试例4：本发明化合物在体外大鼠肝微粒体中的代谢稳定性研究
将待测化合物用DMSO配成10mM母液，并依次用乙腈、PBS稀释，涡匀，控制DMSO含量在0.1％；取10μL稀释好的工作液与65μL肝微粒体体系(30μL 100mM PBS，25μL 20mM MgCl2，10μL 5mg/mL RLM)培养孵育，37℃预热5min；加入25μL 4mM NADPH开启反应；于不同时间点(0min、5min、15min、30min、60min、120min)取样，加入含内标的乙腈溶液终止反应，经涡旋、离心后，收集上清液进行LC-MS/MS分析并计算半衰期和清除率。RLM购于瑞德肝脏。
本发明化合物在体外大鼠肝微粒体中的代谢稳定性研究结果见表4：
表4化合物在体外大鼠肝微粒体中的代谢稳定性研究
图3为本发明化合物A-1、A-3、A-10、A-42在体外大鼠肝微粒体中的代谢稳定性研究结果。
从图表可以看出，化合物A-1、A-3、A-10、A-42的半衰期与临床药物CC-90009相比有着2倍以上的优势，在体外大鼠肝微粒体中具有优秀的代谢稳定性。
生物测试例5：本发明化合物在大鼠体内药代动力学研究
样品采集和准备：生长状态良好的SPF级SD大鼠(雄性，210-230g，42-48天，瑞德肝脏)经静脉2mg/kg和口服10mg/kg给药后，分别于不同采血时间经眼眶采集血样并转移至含抗凝剂的离心管中，其中静脉采血时间为0.08、0.25、0.5、1、2、4、6、8、24小时，口服采血时间为0.25、0.5、1、2、4、6、8、24小时。血样立即在4℃、8000rpm、10分钟条件下离心收集血浆，保存于-80℃超低温冰箱直至LC-MS/MS分析。
药代动力学数据分析：使用药代动力学软件在非室间模型中计算该化合物的药代动力学参数。以血药浓度峰值(Cmax)和血药浓度达峰时间(Tmax)为观察值。通过对数血药浓度-时间曲线的线性末端部分的回归分析以及线性梯形法则，确定半衰期T1/2、药时曲线下面积AUClast和AUCinf、表观分布容积Vss、清除率CL等参数。
本发明化合物在大鼠体内药代动力学研究结果见表5：
表5化合物在大鼠体内药代动力学研究
图4为化合物A-1、A-10、A-42与临床药物CC-90009在大鼠体内药代动力学研究结果。
从图表可以看出，化合物A-1静脉注射给药时，体内清除率小于临床药物CC-90009，半衰期与其接近，在体内具有较好的代谢稳定性，初始血药浓度以及血浆药物暴露量与CC-90009相比具有较大优势。
化合物A-10静脉注射给药的清除率是CC-90009的三分之一，且半衰期长于CC-90009；初始血药浓度远高于CC-90009，血浆药物暴露量接近其四倍；并且口服给药的暴露量接近CC-90009三倍，具有较高的达峰高度和较小的达峰时间，揭示化合物A-10吸收更快。
化合物A-42清除率更低，具有更高的静脉和口服血浆药物暴露量，分别为CC-90009的十四倍和四倍。生物测试例6：本发明化合物在急性髓系白血病异种移植小鼠模型中的体内药效学研究
用于本实验的人源白血病细胞MV411(中科院细胞库)，以IMDM培养基添加10％ FBS培养于含CO2的37℃培养箱。细胞连续培养十代之前，用对数生长期的MV411细胞，用不含血清的培养基调整细胞浓度至1×108个/mL，无菌条件下，通过皮下注射接种于BABL/c Nude小鼠(江苏集萃药康生物科技股份有限公司，雌性，4-6周龄，14-16g)背部右边(靠近腋下)，接种体积为100μL/只。荷瘤小鼠的平均瘤体积达到≈100mm3时，随机分组，各组别肿瘤体积大致相同。分组给药当天定义为第0天。通过口服灌胃15mg/kg、30mg/kg、60mg/kg向每一治疗组中的动物给予化合物(每天一次)。每隔一天测量动物体重及肿瘤体积。
实验指标：化合物的抑瘤疗效用相对肿瘤抑制率TGI评价。瘤重TGI(％)＝{1-[(实验组的最终平均瘤重)/(未给药荷瘤小鼠对照组的最终平均瘤重)]}×100％，瘤体积TGI(％)＝[1-实验组相对肿瘤体积/未给药荷瘤小鼠对照组相对肿瘤体积]*100％。
结果：图5显示了化合物A-10在MV411异种移植小鼠模型中的体内药效学研究。在小鼠体重无明显丢失情况下，化合物A-10在给药剂量分别为15mg/kg、30mg/kg、60mg/kg时，瘤重TGI达到20.82％、56.61％、59.76％，明显优于临床分子CC-90009。图6显示了化合物A-42在MV411异种移植小鼠模型中的体内药效学研究。在小鼠体重无明显丢失情况下，化合物A-42在给药剂量为60mg/kg时，瘤体积TGI达到100％，与临床分子MRT-2359水平相当。
生物测试例7：本发明化合物在前列腺癌异种移植小鼠模型中的体内药效学研究
用于本实验的22RV1细胞(中科院细胞库)，以1640培养基添加10％ FBS培养于含CO2的37℃培养箱。细胞连续培养十代之前，用对数生长期的22RV1细胞，用不含血清的培养基调整细胞浓度至5×107个/mL，无菌条件下，通过皮下注射接种于BABL/c Nude小鼠背部右边(靠近腋下)，接种体积为100μL/只。荷瘤小鼠的平均瘤体积达到≈50-100mm3时，按每组8只，随机分为4个组别，各组别肿瘤体积大致相同。分组给药当天定义为第0天。通过口服灌胃60mg/kg向每一治疗组中的动物给予化合物(每天一次)。每隔一天测量动物体重及肿瘤体积。
实验指标：化合物的抑瘤疗效用相对肿瘤抑制率TGI评价。瘤重TGI(％)＝{1-[(实验组的最终平均瘤重)/(未给药荷瘤小鼠对照组的最终平均瘤重)]}×100％。
结果：图7显示了化合物A-10在22RV1异种移植小鼠模型中的体内药效学研究。化合物A-10在给药剂量为60mg/kg时，TGI达到95.10％，明显优于临床分子CC-90009与上市药物Enzalutamide。
如上所述，尽管参照特定的优选实施例已经表示和表述了本发明，但其不得解释为对本发明自身的限制。在不脱离所附权利要求定义的本发明的精神和范围前提下，可对其在形式上和细节上作出各种变化。

Claims (11)

  1. 一种通式I所示的化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体：
    其中：
    表示单键或双键；
    X选自-CH2-、-NH-、-O-、-S-或-Se-；
    Y选自碳或氮；
    Z选自-CH2-，-CD2-，-C(O)-或-C(S)-；
    每个R1分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    每个R2分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    每个R3分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    R4选自氢、氘、C1-C12烷基、
    R5、R6、R7、R8、R9、R10分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    Ra、Rb分别独立地选自氢、C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、羟基、氰基、硝基、苄基、-C(O)NRcRd、-C(O)Rc、-C(O)ORc、-ORc、-OC(O)Rc、-OC(O)ORc、-OC(O)NRcRd、-NRcRd、-SRc、-S(O)Rc、-S(O)2Rc或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基、杂芳基或苄基未被取代或任选地被1-3个Rc取代；
    Rc、Rd分别独立地选自氢、卤素、羰基、羟基、氰基、硝基、苯基、苄基、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基、C3-C6环烷基或卤代C3-C6环烷基；
    m、n分别独立地选自0、1、2或3。
  2. 一种通式Ⅱ所示的化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体：
    其中：
    表示单键或双键；
    X选自-CH2-、-NH-、-O-、-S-或-Se-；
    Y选自碳或氮；
    Z选自-CH2-，-CD2-，-C(O)-或-C(S)-；
    每个R1分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    每个R2分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    每个R3分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    R4选自氢、氘、C1-C12烷基、
    R5选自氢、氘、C1-C12烷基；
    R6、R7、R8、R9、R10、R11分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基、硝基、-C(O)NRaRb、-C(O)Ra、-C(O)ORa、-ORa、-OC(O)Ra、-OC(O)ORa、-OC(O)NRaRb、-NRaRb、-SRa、-S(O)Ra、-S(O)2Ra或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基和杂芳基未被取代或任选地被1-3个Ra取代；
    Ra、Rb分别独立地选自氢、C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、羟基、氰基、硝基、苄基、-C(O)NRcRd、-C(O)Rc、-C(O)ORc、-ORc、-OC(O)Rc、-OC(O)ORc、-OC(O)NRcRd、-NRcRd、-SRc、-S(O)Rc、-S(O)2Rc或含有0-3个杂原子的3-10元环烷基、杂环基、芳基和杂芳基，所述烷基、烯基、炔基、环烷基、杂环基、芳基、杂芳基或苄基未被取代或任选地被1-3个Rc取代；
    Rc、Rd分别独立地选自氢、卤素、羰基、羟基、氰基、硝基、苯基、苄基、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基、C3-C6环烷基或卤代C3-C6环烷基；
    m、n分别独立地选自0、1、2或3。
  3. 根据权利要求1所述的化合物，其具有式Ia、式Ib或式Ic的结构：
    其中，R1、X、Y、Z、m、n、如通式I中的定义。
  4. 根据权利要求1或2所述的化合物，其特征在于：
    X选自-NH-或-O-。
  5. 根据权利要求1或2所述的化合物，其特征在于：
    每个R1分别独立地选自氢、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基、C2-C6烯基、C2-C6炔基、卤素、氰基、羟基、硝基、C1-C10芳基或-NRaRb；
    Ra、Rb分别独立地选自氢、C1-C6烷基或-C(O)Rc；
    Rc选自氢、卤素、C1-C6烷基、卤代C1-C6烷基、C1-C6烷氧基、卤代C1-C6烷氧基或C3-C6环烷基。
  6. 根据权利要求4所述的化合物，其特征在于：
    每个R1分别独立地选自氢、甲基、氟、氯、甲氧基、三氟甲氧基、硝基、氰基、氨基、羟基、乙烯基、乙炔基、异丁基、苯基、
  7. 根据权利要求1所述的化合物，其特征在于选自：


  8. 一种药物组合物，其特征在于，包括治疗有效量的权利要求1-7任一项所述的化合物或其药学上可接受的盐、互变异构体、内消旋体、外消旋体、对映异构体、非对映异构体以及药学上可接受的载体或赋形剂。
  9. 权利要求1-7任一项所述的化合物在制备用于治疗或预防由GSPT1、IKZF1、IKZF2、IKZF3、CK1α、N-MYC或C-MYC蛋白突变、表达失衡、变构与功能异常相关疾病的药物中的用途。
  10. 根据权利要求9所述的用途，其特征在于，所述相关疾病为癌症、衰老、免疫性疾病、神经性疾病；其中，所述癌症选自急性髓白血病、肝癌、急性淋巴细胞白血病、膀胱癌、骨癌、脑癌、乳腺癌、宫颈癌、绒毛膜癌、慢性淋巴细胞性白血病(CLL)、慢性粒细胞白血病(CML)、结肠癌、食道癌、胆囊癌、胃癌、胃肠道间质瘤、头颈癌、霍奇金淋巴瘤、喉癌、白血病、肺癌、黑色素瘤、间皮瘤、多发性骨髓瘤、卵巢癌、胰腺癌、前列腺癌、直肠癌、肾癌、肉瘤、皮肤癌、小细胞肺癌、睾丸癌、咽喉癌、甲状腺癌或子宫癌。
  11. 权利要求1-7任一项所述的化合物在制备GSPT1降解剂中的用途。
PCT/CN2024/090223 2023-04-28 2024-04-26 一种苯并六元杂环类gspt1蛋白降解剂及其应用 Pending WO2024222918A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202310478561.1 2023-04-28
CN202310478561 2023-04-28

Publications (1)

Publication Number Publication Date
WO2024222918A1 true WO2024222918A1 (zh) 2024-10-31

Family

ID=92237566

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2024/090223 Pending WO2024222918A1 (zh) 2023-04-28 2024-04-26 一种苯并六元杂环类gspt1蛋白降解剂及其应用

Country Status (2)

Country Link
CN (1) CN118496213A (zh)
WO (1) WO2024222918A1 (zh)

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
TW202519521A (zh) * 2023-10-31 2025-05-16 中國藥科大學 一種酞嗪酮類gspt1蛋白降解劑及其應用

Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101291924A (zh) * 2005-08-31 2008-10-22 细胞基因公司 异吲哚-酰亚胺化合物和包含它们的组合物及其使用方法
CN102112463A (zh) * 2008-05-30 2011-06-29 细胞基因公司 5-取代的异二氢吲哚化合物
CN111285850A (zh) * 2018-12-06 2020-06-16 中国科学院上海药物研究所 一类异吲哚啉类化合物、其制备方法、药物组合物及其应用
CN112601751A (zh) * 2018-06-13 2021-04-02 拜欧斯瑞克斯公司 稠合噻吩化合物
CN112745298A (zh) * 2019-10-30 2021-05-04 中国科学院上海药物研究所 多取代异吲哚啉类化合物、其制备方法、药物组合物及用途
CN114085212A (zh) * 2018-09-30 2022-02-25 中国科学院上海药物研究所 异吲哚啉类化合物、其制备方法、药物组合物及用途
CN114502543A (zh) * 2019-05-24 2022-05-13 拜欧斯瑞克斯公司 靶向蛋白化合物、其药物组合物及治疗应用
CN114650868A (zh) * 2019-10-30 2022-06-21 达纳-法伯癌症研究公司 Helios的小分子降解剂及其使用方法
CN114835680A (zh) * 2022-04-29 2022-08-02 成都分迪药业有限公司 卤素取代异吲哚啉化合物及其应用
WO2022257897A1 (en) * 2021-06-08 2022-12-15 Hangzhou Glubio Pharmaceutical Co., Ltd. Isoindolinone compounds, and uses thereof
WO2023274246A1 (zh) * 2021-06-28 2023-01-05 成都分迪药业有限公司 酰胺类化合物及其用途
WO2023085785A1 (ko) * 2021-11-09 2023-05-19 한국화학연구원 글루타이미드 모핵을 갖는 이소인돌리논 유도체 및 이의 용도

Patent Citations (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101291924A (zh) * 2005-08-31 2008-10-22 细胞基因公司 异吲哚-酰亚胺化合物和包含它们的组合物及其使用方法
CN102112463A (zh) * 2008-05-30 2011-06-29 细胞基因公司 5-取代的异二氢吲哚化合物
CN112601751A (zh) * 2018-06-13 2021-04-02 拜欧斯瑞克斯公司 稠合噻吩化合物
CN114085212A (zh) * 2018-09-30 2022-02-25 中国科学院上海药物研究所 异吲哚啉类化合物、其制备方法、药物组合物及用途
CN111285850A (zh) * 2018-12-06 2020-06-16 中国科学院上海药物研究所 一类异吲哚啉类化合物、其制备方法、药物组合物及其应用
CN114502543A (zh) * 2019-05-24 2022-05-13 拜欧斯瑞克斯公司 靶向蛋白化合物、其药物组合物及治疗应用
CN112745298A (zh) * 2019-10-30 2021-05-04 中国科学院上海药物研究所 多取代异吲哚啉类化合物、其制备方法、药物组合物及用途
CN114650868A (zh) * 2019-10-30 2022-06-21 达纳-法伯癌症研究公司 Helios的小分子降解剂及其使用方法
WO2022257897A1 (en) * 2021-06-08 2022-12-15 Hangzhou Glubio Pharmaceutical Co., Ltd. Isoindolinone compounds, and uses thereof
WO2023274246A1 (zh) * 2021-06-28 2023-01-05 成都分迪药业有限公司 酰胺类化合物及其用途
WO2023085785A1 (ko) * 2021-11-09 2023-05-19 한국화학연구원 글루타이미드 모핵을 갖는 이소인돌리논 유도체 및 이의 용도
CN114835680A (zh) * 2022-04-29 2022-08-02 成都分迪药业有限公司 卤素取代异吲哚啉化合物及其应用

Also Published As

Publication number Publication date
CN118496213A (zh) 2024-08-16

Similar Documents

Publication Publication Date Title
US9890168B2 (en) 2,4-disubstituted 7H-pyrrolo[2,3-d]pyrimidine derivative, preparation method and medicinal use thereof
CN104045642B (zh) 含嘧啶或吡啶的稠环化合物及其作为抗肿瘤药物的应用
CN112672997A (zh) 哒嗪酮衍生物
US10196358B2 (en) KCNQ2-5 channel activator
JP6054402B2 (ja) ＨＩＦ−１α活性を阻害する新規化合物及びその製造方法
CN106488918B (zh) 三唑并嘧啶酮或三唑并吡啶酮衍生物及其用途
AU2021400725A9 (en) Compounds useful as t cell activators
CN107922348A (zh) 双环杂环酰胺衍生物
JP6545262B2 (ja) Ｒｉｐｋ２阻害剤およびそれを用いた癌の治療方法
WO2016050165A1 (zh) 氮杂双环衍生物、其制法与医药上的用途
CN109535164B (zh) Jak激酶抑制剂及其制备方法和在医药领域的应用
WO2016117647A1 (ja) 新規ベンズイミダゾール誘導体およびその医薬用途
WO2024222918A1 (zh) 一种苯并六元杂环类gspt1蛋白降解剂及其应用
WO2019085808A1 (zh) 一种含三氮唑的ido抑制剂、其制备方法及其医药用途
CN111978325B (zh) 咪唑并哒嗪类mnk1/mnk2激酶抑制剂及其制备方法和应用
CN103509024B (zh) 嘧啶并苯并氮杂卓类化合物及其作为抗肿瘤药物的应用
CN112996507B (zh) 肥胖症的治疗
CN103626761B (zh) 苯并吡啶氮杂卓类化合物及其作为抗肿瘤药物的应用
WO2025092775A1 (zh) 一种酞嗪酮类gspt1蛋白降解剂及其应用
CN114805371B (zh) 含2-氨基嘧啶大环类化合物及其制备方法和用途
WO2024042163A1 (en) Novel substituted 2,3,4,9-tetrahydro-1h-pyrido[3,4-b]indole carboxylic acid derivatives, processes for their preparation and therapeutic uses thereof
WO2024251301A1 (zh) 一种含肟结构的gspt1蛋白降解剂及其制备方法和应用
CN108117551A (zh) 取代(1H-吡唑[3,4-b]吡啶)脲类化合物及其抗肿瘤用途
WO2024256574A1 (en) Process for preparing macrocyclic btk inhibitors
CN120040418A (zh) 一类含有单环芳基取代的cdk12/13的降解剂，及其制备方法、药物组合物和应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 24796287

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 2024796287

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2024796287

Country of ref document: EP

Effective date: 20251128

ENP Entry into the national phase

Ref document number: 2024796287

Country of ref document: EP

Effective date: 20251128

ENP Entry into the national phase

Ref document number: 2024796287

Country of ref document: EP

Effective date: 20251128