WO2024251301A1 - 一种含肟结构的gspt1蛋白降解剂及其制备方法和应用 - Google Patents

一种含肟结构的gspt1蛋白降解剂及其制备方法和应用 Download PDF

Info

Publication number
WO2024251301A1
WO2024251301A1 PCT/CN2024/103853 CN2024103853W WO2024251301A1 WO 2024251301 A1 WO2024251301 A1 WO 2024251301A1 CN 2024103853 W CN2024103853 W CN 2024103853W WO 2024251301 A1 WO2024251301 A1 WO 2024251301A1
Authority
WO
WIPO (PCT)
Prior art keywords
cancer
alkyl
compound
hydrogen
nrarb
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.)
Ceased
Application number
PCT/CN2024/103853
Other languages
English (en)
French (fr)
Inventor
李志裕
徐熙
瞿凡贵
常秀锦
范忠蓬
张艳青
卞金磊
王举波
邱志霞
吴红茜
童超
马萌阳
李春晓
陈冉冉
陈鑫
邹昕羽
孙锋泽
张忠霖
张若萱
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
Original Assignee
China Pharmaceutical University
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 filed Critical China Pharmaceutical University
Publication of WO2024251301A1 publication Critical patent/WO2024251301A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A61K31/405—Indole-alkanecarboxylic acids; Derivatives thereof, e.g. tryptophan, indomethacin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/445—Non condensed piperidines, e.g. piperocaine
    • A61K31/4523—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems
    • A61K31/454—Non condensed piperidines, e.g. piperocaine containing further heterocyclic ring systems containing a five-membered ring with nitrogen as a ring hetero atom, e.g. pimozide, domperidone
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/06—Immunosuppressants, e.g. drugs for graft rejection
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P39/00—General protective or antinoxious agents
    • A61P39/06—Free radical scavengers or antioxidants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Definitions

  • the present invention belongs to the technical field of chemical medicine, and in particular relates to a GSPT1 protein degradation agent containing an oxime structure, 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
  • Targeted protein degradation technology has broad application prospects and development space in a variety of disease fields, such as malignant tumors, neurodegenerative diseases, metabolic diseases and other fields.
  • 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
  • CC-885 as a 5-substituted isoindoline compound, has strong CRBN-dependent antiproliferative activity in various tumor cell lines and has enhanced antiproliferative activity compared with lenalidomide and pomalidomide (Nature 2016, 535, 252-257). Immunoprecipitation assays and immunoblotting assays showed that CC-885 promoted the binding of CRBN to the novel substrate GSPT1, targeting its degradation. Using CC-885 as a structural template, a skeleton with difluoroacetamide was discovered through SAR, which showed good selectivity in vitro (J.Med.Chem.2021, 64, 1835-1843).
  • CC-90009 is a highly efficient GSPT1 selective degrader that shows good PK/PD and safety in the treatment of acute myeloid leukemia and has entered Phase I/II clinical trials (NCT04297124; NCT02848001; NCT04336982).
  • 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.
  • 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.
  • the purpose of the present invention is to provide a GSPT1 protein degradation agent containing an oxime structure and its preparation method and application.
  • the constructed compounds have excellent GSPT1 degradation activity and can be used as GSPT1 protein degraders. They broaden the skeleton structure of GSPT1 molecular glue degraders, enhance the anti-proliferative effect on tumors, improve the therapeutic window, and provide more options for the research and development of GSPT1 molecular glue.
  • the present invention provides a compound represented by general formula I or a pharmaceutically acceptable salt, tautomer, mesomer, racemate, enantiomer, diastereomer thereof:
  • X is selected from -OH, -NH 2 , -OCH 3 or -NHCOCH 3 ;
  • Ring Y is selected from C 6 -C 12 aryl, 5-10 membered heteroaryl, C 3 -C 8 cycloalkyl or 3-10 membered heterocyclyl, wherein the aryl, heteroaryl, cycloalkyl and heterocyclyl are unsubstituted or optionally substituted with 1-3 Ra ;
  • R 1 is selected from
  • R is 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, wherein the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are unsubstituted or optionally substituted with 1-3
  • 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
  • R 4 is selected from hydrogen, deuterium, halogen or C 1 -C 6 alkyl
  • R5 is selected from hydrogen, deuterium, C1 - C12 alkyl
  • Z is selected from -CH2- , -CD2- , -C(O)- or -C(S)-;
  • 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
  • each Ra and Rb are 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, and the alkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl, heteroaryl or benzyl is unsubstitute
  • Each R c and R d is 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 p are independently selected from 0, 1, 2 or 3.
  • X is selected from -OH, -NH 2 or -OCH 3 .
  • R2 is 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 ;
  • 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;
  • Ring Y is selected from C 6 -C 12 aryl or 5-10 membered heteroaryl.
  • R 3 is 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 or nitro;
  • R 4 is selected from hydrogen, deuterium, halogen or C 1 -C 6 alkyl
  • R 5 is selected from hydrogen, deuterium or C 1 -C 12 alkyl
  • n and n are independently selected from 1;
  • p is selected from 0 or 1.
  • 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:
  • the present invention also provides a method for preparing compounds of formula Ia, Ib, and Ic.
  • the synthetic route is as follows:
  • the starting material 1 is brominated with N-bromosuccinimide in the presence of a free radical initiator to obtain an intermediate 2; intermediate 2 is substituted with a nucleophilic reagent 3-aminopiperidine-2,6-dione hydrochloride and further formed into a lactam on the ester to obtain an intermediate 3; a cyano group is inserted into the aromatic bromide through palladium mediation to obtain an intermediate 4; followed by reduction and de-tert-butyloxycarbonylation to obtain an intermediate 6, and finally in the presence of a condensation agent EDCI, compounds related to formulas Ia, Ib, and Ic are obtained.
  • Another object of the present invention is to provide a pharmaceutical composition, which comprises a therapeutically effective amount of a compound of formula I 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 a compound of formula I or a pharmaceutically acceptable salt, tautomer, mesomer, racemate, enantiomer, diastereomer thereof for use in the preparation of a medicament 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, viral infection, 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
  • the present invention also provides the use of the compound of general formula I or its pharmaceutically acceptable salt, tautomer, mesomer, racemate, enantiomer, diastereomer in the preparation of GSPT1 degradation agent.
  • 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 pairs of isomers. 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 invention 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.
  • the oxime compounds of the present invention have significantly improved degradation effects on GSPT1 protein and anti-proliferation inhibitory activity of AML cells.
  • 200nM compound OX-7 was administered to KG-1 cells for 4 hours, and the degradation percentage of GSPT1 protein was higher than 80%. Under the same conditions, the degradation percentage of GSPT1 protein by compound NX-7 was higher than 90%.
  • compounds OX-7 and NX-7 can also significantly degrade C-MYC protein; compound OX-7 can also significantly induce the production of c-PARP protein, has a significant effect of inducing cell apoptosis, and broadens the scope of tumor indications.
  • compound OX-7 began to have a significant degradation effect on GSPT1 and C-MYC at 2h, and as time goes by, the degradation effect is better, showing a significant time dependence.
  • 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.
  • Figure 1 shows that the degradation effect of the compound OX-7 of the present invention on GSPT1 in AML cells is concentration-dependent.
  • Figure 1A shows the degradation effect of the compound OX-7 and the clinical drug CC-90009 on GSPT1 in KG-1 cells;
  • Figure 1B shows the degradation effect of the compound OX-7 and the clinical drug CC-90009 on GSPT1 in MOLM-13 cells.
  • FIG. 2 shows that the degradation effect of the compound OX-7 of the present invention on GSPT1 in AML cells is time-dependent.
  • Figure 3 shows the antiproliferative activity of the compound OX-7 of the present invention and the clinical drug CC-90009 on different tumor cells.
  • Figures 3A and 3B show the antiproliferative activity of OX-7 and CC-90009 on KG-1 cells, respectively;
  • Figures 3C and 3D show the antiproliferative activity of OX-7 and CC-90009 on MOLM-13 cells, respectively.
  • 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.
  • the reaction is complete as monitored by TLC, and the reaction solution is poured into 200 mL of water to precipitate a light blue solid, which is allowed to stand for complete precipitation, and then filtered.
  • the filter cake is vacuum dried to obtain a light blue solid 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 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 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 can be directly used in the next step without further purification.
  • Step 1 Benzaldehyde (2.00 g, 18.86 mmol) was dissolved in 20 mL of acetic anhydride, and N-acetylglycine (3.31 g, 28.29 mmol) and sodium acetate (2.32 g, 28.29 mmol) were added in sequence, and the mixture was heated to 120°C and stirred for 5 hours.
  • Step 2 (Z)-4-benzylidene-2-methyloxazol-5(4H)-one (2.20 g, 11.76 mmol) was dissolved in 50 mL of 10% hydrochloric acid aqueous solution, heated to 100°C and stirred for 10 hours. The reaction was complete as monitored by TLC, and the reaction solution was cooled to room temperature. A brown solid was precipitated under ice bath, and the precipitation was complete after standing. The solid was filtered and the filter cake was dried under vacuum to obtain a brown solid, which was intermediate-2: 2-oxo-3-phenylpropionic acid (1.52 g, yield 78.7%). The product can be used directly in the next step without further purification.
  • Step 3 2-oxo-3-phenylpropionic acid (1.52 g, 9.27 mmol) was dissolved in 20 mL of pyridine, and O-(tetrahydro-2H-pyran-2-yl)hydroxylamine (1.30 g, 11.12 mmol) was added, and stirred at room temperature for 12 hours under nitrogen protection.
  • Step 1 2-oxo-3-phenylpropionic acid (1.52 g, 9.26 mmol) was dissolved in 20 mL of pyridine, methoxyamine hydrochloride (1.16 g, 13.89 mmol) was added, and the mixture was stirred at room temperature for 12 hours under nitrogen protection.
  • Step 1 Dissolve 5-nitroisobenzofuran-1,3-dione (14.50 g, 75.00 mmol) in 20 mL of acetic acid solution, add 3-aminopiperidine-2,6-dione hydrochloride (8.20 g, 50.00 mmol) and sodium acetate (5.00 g, 60.00 mmol) in turn, heat to reflux and stir for 5 h.
  • the reaction is complete as monitored by TLC. After the reaction solution is cooled to room temperature, it is poured into 100 mL of ice water to precipitate a white solid, which is allowed to stand for complete precipitation.
  • Step 2 Dissolve 2-(2,6-dioxopiperidin-3-yl)-5-nitroisoindoline-1,3-dione (3.63 g, 11.98 mmol) in 10 mL of N,N-dimethylformamide, add 10% palladium carbon (0.54 g, 5.07 mmol), and stir for 12 hours at room temperature under full replacement of hydrogen. The reaction is complete as monitored by TLC, and the filtrate is filtered and poured into 100 mL of ice water to precipitate a yellow solid.
  • 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.
  • Step 1 Dissolve (E)-3-phenyl-2-(((tetrahydro-2H-pyran-2-yl)oxy)imino)propionic acid (Intermediate-3, 2.00 g, 7.60 mmol) in 10 mL of N,N-dimethylformamide, add EDCI (2.18 g, 11.40 mmol) and HOBT (1.54 g, 11.40 mmol) in turn, stir at room temperature for 1 hour after addition. Add 3-(5-aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (1.17 g, 3.80 mmol) and stir at room temperature overnight. The reaction was completed as monitored by TLC.
  • Step 2 Compound 10 (0.50 g, 0.96 mmol) was dissolved in 10 mL of hydrochloric acid-dioxane solution (4 M) and stirred at room temperature for 3 hours. The reaction was completed as monitored by TLC, and the mixture was filtered and the filter cake was purified by methanol slurry and vacuum dried to obtain a white solid OX-1 (0.20 g, yield 47.6%, purity 98.61%).
  • Step 1 (E)-3-(3-chloro-4-methylphenyl)-2-(((tetrahydro-2H-pyran-2-yl)oxy)imino)propanoic acid (Compound 21, 1.25 g, 4.03 mmol) was dissolved in 10 mL of N,N-dimethylformamide, and EDCI (1.05 g, 5.49 mmol) and HOBT (0.74 g, 5.49 mmol) were added in sequence. After the addition was completed, the mixture was stirred at room temperature for 1 hour.
  • Step 1 Dissolve 2-oxo-3-phenylpropionic acid (Intermediate-2, 1.06 g, 6.47 mmol) in 10 mL of N,N-dimethylformamide, add EDCI (1.24 g, 6.47 mmol) and HOBT (0.87 g, 6.47 mmol) in turn, stir at room temperature for 1 hour after addition. Add 3-(5-aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (1.00 g, 3.23 mmol) and stir at room temperature overnight. The reaction was completed as monitored by TLC, and the reaction solution was poured into 50 mL of water and extracted with ethyl acetate.
  • Step 2 Compound 32 (0.50 g, 1.15 mmol) was dissolved in 10 mL of acetic acid solution (50%), 80% hydrazine hydrate (0.07 g, 1.15 mmol) was added, and the mixture was stirred at room temperature for 5 h. The reaction was completed as monitored by TLC. The reaction solution was poured into 50 mL of saturated sodium bicarbonate solution, extracted with ethyl acetate, and the organic phases were combined and washed with water and saturated brine in sequence. After drying over anhydrous sodium sulfate, the mixture was concentrated in vacuo.
  • Step 1 Dissolve (E)-2-(2-aminothiazol-4-yl)-2-(methoxyimino)acetic acid (compound 54, 0.5 g, 2.48 mmol) in a mixed solvent of 4 mL of N,N-dimethylformamide and 10 mL of tetrahydrofuran, add dropwise 4 mL of a tetrahydrofuran solution of sodium bis(trimethylsilyl)amide (2 M) under ice bath, stir under ice bath for 1 h, add dropwise di-tert-butyl dicarbonate (0.58 g, 2.68 mmol) at 0°C, stir under room temperature for 2 h.
  • Step 2 Dissolve 3-(5-aminomethyl)-1-oxoisoindolin-2-yl)piperidine-2,6-dione hydrochloride (Intermediate-1, 0.15 g, 0.49 mmol) in 6 mL of N,N-dimethylformamide, add compound 55 (0.15 g, 0.49 mmol) and HATU (0.28 g, 0.74 mmol) in turn, stir at room temperature for 1 hour after addition, add DIPEA (0.19 g, 1.47 mmol), and stir at room temperature overnight. The reaction was completed as monitored by TLC. The reaction solution was poured into 30 mL of ice water to precipitate a brown solid. The solution was allowed to stand for complete precipitation and filtered.
  • Step 3 Compound 56 (0.11 g, 0.19 mmol) was dissolved in 30 mL of hydrochloric acid-dioxane solution (4 M) and stirred at room temperature for 3 hours. The reaction was completed as monitored by TLC, and the filter cake was vacuum-dried to obtain a light yellow solid CX-11 (0.04 g, yield 45.9%, purity 98.87%).
  • Biological test example 1 Study on the degradation of GSPT1 in KG-1 cells.
  • 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.
  • the control group was a blank solvent group.
  • 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 degradation effect of the dominant compounds in each series of compounds of the present invention is better than that of the clinical drug, and the degradation effect of most other compounds is comparable to that of the clinical drug. It can be seen that the degradation effect of the oxime compounds OX, NX, and CX series on GSPT1 protein can be significantly improved.
  • the present invention takes compound OX-7 as an example, and provides a graph showing that its degradation effect on GSPT1 in AML cells is concentration-dependent and time-dependent, as shown in Figures 1 and 2.
  • Figure 1A shows the degradation effect of compound OX-7 and clinical drug CC-90009 on GSPT1 in KG-1 cells.
  • Figure 1B shows the degradation effect of compound OX-7 and clinical drug CC-90009 on GSPT1 in MOLM-13 cells.
  • the degradation effect of the compound OX-7 of the present invention on GSPT1 in KG-1 and MOLM-13 cells is concentration-dependent and time-dependent, and the effect is more significant than that of CC-90009.
  • Biological test example 2 Anti-KG-1 cell proliferation activity of the compounds of the present invention.
  • 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 antiproliferative inhibitory activity of the compounds OX, NX, and CX series of the present invention on KG-1 cells is significantly improved, which is better than the clinical drug CC-90009.
  • the superior compounds of each series such as OX-7, NX-7, CX-7, and CX-8 have antiproliferative effects. It has excellent proliferation inhibitory activity, with IC50 ranging from 1-10 nM.
  • Biological test example 3 The compounds of the present invention have anti-proliferative activity against a variety of 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 antiproliferative inhibitory activity of the dominant compounds in each series of compounds of the present invention is better than CC-90009, and the other compounds are also comparable to CC-90009. Therefore, it can be seen that the antiproliferative inhibitory activity of the oxime compounds OX, NX, and CX series of the present invention on various AML cells is significantly improved.

Landscapes

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Epidemiology (AREA)
  • Oncology (AREA)
  • Toxicology (AREA)
  • Communicable Diseases (AREA)
  • Biomedical Technology (AREA)
  • Neurology (AREA)
  • Neurosurgery (AREA)
  • Biochemistry (AREA)
  • Transplantation (AREA)
  • Hematology (AREA)
  • Virology (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

一种含肟结构的GSPT1蛋白降解剂及其制备方法和应用，本发明公开了一种含肟结构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。本发明的化合物具有优良的GSPT1降解活性，可以作为GSPT1蛋白降解剂，拓宽了GSPT1分子胶降解剂的骨架结构，提高了对肿瘤的抗增殖作用，改善治疗窗，为GSPT1分子胶的研发提供更多的选择。与临床药物CC-90009相比，本发明的肟类化合物如OX、NX、CX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。

Description

一种含肟结构的GSPT1蛋白降解剂及其制备方法和应用 技术领域
本发明属于化学医药技术领域，具体涉及一种含肟结构的GSPT1蛋白降解剂及其制备方法和应用。
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多种疾病领域具有广阔的应用前景和发展空间，如恶性肿瘤、神经退行性疾病、代谢性疾病等领域。
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。
CC-885作为5-取代异吲哚啉化合物，在各类肿瘤细胞系中广泛具有强CRBN依赖性抗增殖活性，与来那度胺和泊马度胺相比具有增强的抗增殖活性(Nature 2016,535,252-257)。免疫沉淀测定和免疫印迹测定表明，CC-885促进CRBN与新型底物GSPT1的结合，靶向其降解。使用CC-885作为结构模板，通过SAR发现了一种具有二氟乙酰胺的骨架，在体外显示良好的选择性(J.Med.Chem.2021,64,1835-1843)。从这个基础上，进一步筛选出BMS/Celgene的第三代先导化合物CC-90009。CC-90009是一种高效的GSPT1选择性降解剂，在治疗急性髓系白血病方面表现出良好的PK/PD和安全性，目前已进入Ⅰ/Ⅱ期临床试验(NCT04297124；NCT02848001；NCT04336982)。
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。
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：本发明目的在于提供一种含肟结构的GSPT1蛋白降解剂及其制备方法和应用。本发明含肟结 构的化合物具有优良的GSPT1降解活性，可以作为GSPT1蛋白降解剂，拓宽了GSPT1分子胶降解剂的骨架结构，提高了对肿瘤的抗增殖作用，改善治疗窗，为GSPT1分子胶的研发提供更多的选择。
技术方案：本发明的目的通过下述技术方案实现：
本发明提供了一种通式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：
X选自-OH、-NH2、-OCH3或-NHCOCH3；
环Y选自C6-C12芳基、5-10元杂芳基、C3-C8环烷基或3-10元杂环基，所述芳基、杂芳基、环烷基和杂环基未被取代或任选地被1-3个Ra取代；
R1选自
R2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、卤素或C1-C6烷基；
R5选自氢、氘、C1-C12烷基、
Z选自-CH2-，-CD2-，-C(O)-或-C(S)-；
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；
每个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；
每个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、p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，X、Y、Z、m、n、p如通式I中所定义。
在某些优选的实施方式中，
X选自-OH、-NH2或-OCH3。
在某些优选的实施方式中，
R2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；
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；
环Y选自C6-C12芳基或5-10元杂芳基。
在某些更优选的实施方式中，
R3选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基或硝基；
R4选自氢、氘、卤素或C1-C6烷基；
R5选自氢、氘或C1-C12烷基；
m、n分别独立地选自1；
p选自0或1。
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：


本发明还提供了式Ia、Ib、Ic相关化合物的制备方法，合成路线如下：
起始原料1在自由基引发剂存在下，用N-溴代丁二酰亚胺溴化，得到中间体2；中间体2与亲核试剂3-氨基哌啶-2,6-二酮盐酸盐进行取代后，在酯上进一步成内酰胺，得到中间体3；通过钯介导将氰基插入到芳基溴上，得到中间体4；随后还原、脱叔丁氧羰基得到中间体6，最后在缩合剂EDCI的存在下，得到式Ia、Ib、Ic相关化合物。
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、病毒感染、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。
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”是指本发明化合物的以下量，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。
有益效果：
与临床药物CC-90009相比，本发明的肟类化合物如OX、NX、CX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OX-7在KG-1细胞给药4小时，对GSPT1蛋白的降解百分数高于80％。化合物NX-7在相同条件下，对GSPT1蛋白的降解百分数高于90％。与临床药物CC-90009相比，化合物OX-7和NX-7还能显著降解C-MYC蛋白；化合物OX-7还能够显著诱导c-PARP蛋白的产生，具有明显的诱导细胞凋亡的作用，拓宽了肿瘤适应症的范围。在给药浓度100nM时化合物OX-7在2h对GSPT1和C-MYC开始有明显的降解效果，且随着时间的延长，降解效果越好，呈现出明显的时间依赖性。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。
附图说明
图1为本发明化合物OX-7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OX-7与临床药物CC-90009在KG-1细胞中对GSPT1的降解作用；图1B为化合物OX-7与临床药物CC-90009在MOLM-13细胞中对GSPT1的降解作用。
图2为本发明化合物OX-7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OX-7以及临床药物CC-90009对不同肿瘤细胞的抗增殖抑制活性。其中图3A、3B分别为OX-7、CC-90009对KG-1细胞的抗增殖抑制活性；图3C、3D分别为OX-7、CC-90009对MOLM-13细胞的抗增殖抑制活性。
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、中间体-3的合成：
步骤1：将苯甲醛(2.00g，18.86mmol)溶于20mL的乙酸酐中，依次加入N-乙酰甘氨酸(3.31g，28.29mmol)、乙酸钠(2.32g，28.29mmol)加毕，升温至120℃条件下搅拌5小时。经TLC监测反应完全，将反应液冷却至室温，析出橙黄色固体，冰浴下静置使其析出完全，抽滤，滤饼真空干燥得黄色固体7：(Z)-4-亚苄基-2-甲基恶唑-5(4H)-酮(2.2g，收率62.5％)，该产物无需进一步纯化可直接用于下一步。
HRMS(ESI+):计算值C11H10NO2(M+H)+,188.0706；实测值188.0718。
步骤2：将(Z)-4-亚苄基-2-甲基恶唑-5(4H)-酮(2.20g，11.76mmol)溶于50mL的10％盐酸水溶液中，升温至100℃条件下搅拌10小时。经TLC监测反应完全，将反应液冷却至室温，冰浴下析出棕色固体，静置使其析出完全，抽滤，滤饼真空干燥得棕黄色固体，即为中间体-2：2-氧代-3-苯基丙酸(1.52g，收率78.7％)。该产物无需进一步纯化可直接用于下一步。
HRMS(ESI-):计算值C9H7O3(M-H)-,163.0401；实测值163.0412。
步骤3：将2-氧代-3-苯基丙酸(1.52g，9.27mmol)溶于20mL的吡啶中，加入O-(四氢-2H-吡喃-2-基)羟胺(1.30g，11.12mmol)加毕，于氮气保护下室温搅拌12小时。经TLC监测反应完全，将反应液倒入1N的盐酸溶液中，使用乙酸乙酯萃取，合并有机相使用饱和食盐水洗涤，无水硫酸钠干燥后，真空浓缩得到棕褐色固体，即为中间体-3：(E)-3-苯基-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸(1.95g，收率80.0％)。
HRMS(ESI-):计算值C14H16NO4(M-H)-,262.1085；实测值262.1079。
关键中间体-4的合成：
步骤1：将2-氧代-3-苯基丙酸(1.52g，9.26mmol)溶于20mL的吡啶中，加入甲氧胺盐酸盐(1.16g，13.89mmol)加毕，于氮气保护下室温搅拌12小时。经TLC监测反应完全，将反应液倒入1N的盐酸溶液中，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，真空浓缩得到棕褐色固体，即为中间体-4：(E)-2-(甲氧基亚氨基)-3-苯基丙酸(1.21g，收率67.9％)。
HRMS(ESI-):计算值C10H10NO3(M-H)-,192.0666；实测值192.0678。
关键中间体-5的合成：
步骤1：将5-硝基异苯并呋喃-1,3-二酮(14.50g，75.00mmol)溶于20mL的乙酸溶液中，依次加入3-氨基哌啶-2,6-二酮盐酸盐(8.20g，50.00mmol)、乙酸钠(5.00g，60.00mmol)升温至回流条件下搅拌5h。经TLC监测反应完全，将反应液冷却至室温后，倒入100mL的冰水中，析出白色固体，静置使其析出完全， 抽滤，滤饼真空干燥得到灰白色固体9：2-(2,6-二氧哌啶-3-基)-5-硝基异吲哚啉-1,3-二酮(9.03g，收率62.4％)。HRMS(ESI+):计算值C13H10N3O6(M+H)+,304.0564；实测值304.0575。
步骤2：将2-(2,6-二氧哌啶-3-基)-5-硝基异吲哚啉-1,3-二酮(3.63g，11.98mmol)溶于10mL的N,N-二甲基甲酰胺中，加入10％的钯碳(0.54g，5.07mmol)，加毕，于氢气充分置换下，室温条件下搅拌12小时。经TLC监测反应完全，抽滤，将滤液倒入100mL冰水中，析出黄色固体，静置使其析出完全，抽滤，滤饼真空干燥得到黄色固体，即为中间体5：5-氨基-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮(3.00g，收率92.0％)。HRMS(ESI+):计算值C13H12N3O4(M+H)+,274.0822；计算值274.0834。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 化合物OX-1：(E)-N-(2-(2，6-二氧哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟亚胺基)-3-苯基丙酰胺的合成：
步骤1：将(E)-3-苯基-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸(中间体-3，2.00g，7.60mmol)溶解于10mL的N,N-二甲基甲酰胺中，依次加入EDCI(2.18g，11.40mmol)、HOBT(1.54g，11.40mmol)，加毕，室温搅拌1小时。加入3-(5-氨甲基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮盐酸盐(1.17g，3.80mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入30mL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10：(E)-N-(2-(2,6-二氧哌啶-3-基)-1-氧代异吲哚-5-基)甲基)-3-苯基-2-((四氢-2H-吡喃-2-基)氧基)亚胺)丙酰胺(1.01g，收率51.0％)。
步骤2：将化合物10(0.50g，0.96mmol)溶于10mL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，滤饼用甲醇打浆纯化后真空干燥得到白色固体OX-1(0.20g，收率47.6％，纯度98.61％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.96(s,1H),11.02(s,1H),8.70(t,J＝6.3Hz,1H),7.82(d,J＝7.7Hz,1H),7.45(s,1H),7.35-7.20(m,5H),7.13(s,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.50-4.21(m,4H),3.85(s,2H),3.00-2.83(m,1H),2.60(d,J＝17.6Hz,1H),2.47-2.32(m,1H),2.07-1.96(m,1H)。
HRMS(ESI+):计算值C23H23N4O5(M+H)+,435.1663；实测值435.1658。
实施例2 化合物OX-2：(E)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-3-(4-氟苯基)-2-(羟基亚氨基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为对氟苯甲醛，制得化合物11：(E)-3-(4-氟苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸。三步收率共计73.7％。
HRMS(ESI-):计算值C14H15FNO4(M-H)-,280.0991；实测值280.0987。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物11，制得化合物12；将化合物10替换为化合物12，制得白色固体OX-2：(0.18g，收率42.8％，纯度99.40％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.95(s,1H),11.02(s,1H),8.70(t,J＝6.3Hz,1H),7.66(d,J＝7.7Hz,1H),7.39(s,1H),7.35 (d,J＝7.8Hz,1H),7.31-7.18(m,2H),7.16-7.04(m,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.50-4.21(m,4H),3.82(s,2H),3.02-2.83(m,1H),2.60(d,J＝18.0Hz,1H),2.47-2.31(m,1H),2.07-1.94(m,1H)。
HRMS(ESI+):计算值C23H22FN4O5(M+H)+,453.1569；实测值453.1566。
实施例3 化合物OX-3：(E)-3-(4-氯苯基)-N-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为对氯苯甲醛，制得化合物13：(E)-3-(4-氯苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸。三步收率共计49.6％。HRMS(ESI-):计算值C14H15ClNO4(M-H)-,296.0695；实测值296.0688。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物13，制得化合物14：将化合物10替换为化合物14，制得白色固体OX-3(0.15g，收率35.7％，纯度99.02％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.99(s,1H),11.02(s,1H),8.71(t,J＝6.3Hz,1H),7.66(d,J＝7.7Hz,1H),7.39-7.31(m,4H),7.23(d,J＝8.3Hz,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.50-4.20(m,4H),3.83(s,2H),3.01-2.84(m,1H),2.60(d,J＝17.3Hz,1H),2.48-2.29(m,1H),2.06-1.94(m,1H)。
HRMS(ESI+):计算值C23H22ClN4O5(M+H)+,469.1273；实测值469.1268。
实施例4 化合物OX-4：(E)-3-(4-溴苯基)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为对溴苯甲醛，制得化合物15：(E)-3-(4-溴苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸。三步收率共计40.3％。HRMS(ESI-):计算值C14H15BrNO4(M-H)-,340.0190；实测值340.0186。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物15，制得化合物16：将化合物10替换为化合物16，制得白色固体OX-4(0.17g，收率39.5％，纯度98.97％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.97(s,1H),11.00(s,1H),8.68(t,J＝6.3Hz,1H),7.66(d,J＝7.7Hz,1H),7.47(d,J＝8.4Hz,2H),7.35(d,J＝8.4Hz,2H),7.17(d,J＝8.3Hz,2H),5.11(dd,J＝13.2,5.0Hz,1H),4.50-4.18(m,4H),3.81(s,2H),3.02-2.83(m,1H),2.60(d,J＝17.6Hz,1H),2.44-2.29(m,1H),2.08-1.93(m,1H)。HRMS(ESI+):计算值C23H22BrN4O5(M+H)+,513.0768；实测值513.0767。
实施例5 化合物OX-5：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)-3-(4-硝基苯基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为对硝基苯甲醛，制得化合物17：(E)-3-(4-硝基苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸。三步收率共计51.3％。HRMS(ESI-):计算值C14H15N2O6(M-H)-,307.0936；实测值307.0929。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物17，制得化合物18：将化合物10替换为化合物18，制得白色固体OX-5(0.14g，收率32.5％，纯度96.83％)。1H NMR(300MHz,DMSO-d6)δ(ppm)12.12(s,1H),10.99(s,1H),8.74(t,J＝6.3Hz,1H),8.16(d,J＝8.7Hz,2H),7.66(d,J＝7.8Hz,1H),7.48(d,J＝8.6Hz,2H),7.41(s,1H),7.37(d,J＝7.8Hz,1H),5.10(dd,J＝13.2,5.0Hz,1H),4.51-4.21(m,4H),3.97(s,2H),3.00-2.89(m,1H),2.60(d,J＝16.9Hz,1H),2.46-2.28(m,1H),2.05-1.95(m,1H)。HRMS(ESI+):计算值C23H22N5O7(M+H)+,480.1514；实测值480.1510。
实施例6 化合物OX-6：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)-3-(4-吗啉苯基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为4-(4-吗啉)苯甲醛，制得化合物19：(E)-3-(4-吗啉苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸。三步收率共计47.6％。HRMS(ESI-):计算值C18H23N2O5(M-H)-,347.1612；实测值347.1621。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物19，制得化合物20；化合物10替换为化合物20，制得白色固体OX-6(0.15g，收率34.8％，纯度95.80％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.79(s,1H),11.00(s,1H),8.62(t,J＝6.3Hz,1H),7.65(d,J＝7.8Hz,1H),7.38(s,1H),7.33(d,J＝7.9Hz,1H),7.09(s,1H),7.06(s,1H),6.85(s,1H),6.82(s,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.49-4.20(m,4H),3.77-3.68(m,6H),3.03(t,J＝4.8Hz,4H),2.98-2.83(m,1H),2.60(d,J＝17.1Hz,1H),2.45-2.28(m,1H),2.04-1.93(m,1H)。
HRMS(ESI+):计算值C27H30N5O6(M+H)+,520.2191；实测值520.2185。
实施例7 化合物OX-7：(E)-3-(3-氯-4-甲基苯基)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为3-氯-4-甲基苯甲醛，制得化合物21：(E)-3-(3-氯-4-甲基苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸。三步收率共计49.6％。
HRMS(ESI-):计算值C15H17ClNO4(M-H)-,310.0852；实测值310.0847。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物21，制得化合物22；化合物10替换为化合物22，制得白色固体OX-7(0.17g，收率37.2％，纯度99.05％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.96(s,1H),10.99(s,1H),8.69(t,J＝6.3Hz,1H),7.65(d,J＝7.8Hz,1H),7.40(s,1H),7.35(d,J＝7.8Hz,1H),7.24(d,J＝7.8Hz,1H),7.22(s,1H),7.07(d,J＝7.9,1.8Hz,1H),5.11(dd,J＝13.3,5.1Hz,1H),4.49-4.21(m,4H),3.80(s,2H),3.01-2.83(m,1H),2.60(d,J＝17.4Hz,1H),2.47-2.31(m,1H),2.27(s,3H),2.05-1.98(m,1H)。
HRMS(ESI+):计算值C24H24ClN4O5(M+H)+,483.1430；实测值483.1426。
实施例8 化合物OX-8：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)-3-(4-(三氟甲氧基)苯基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为对三氟甲氧基苯甲醛，制得化合物23：(E)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)-3-(4-(三氟甲氧基)苯基)丙酸。三步收率共计51.3％。
HRMS(ESI-):计算值C15H15F3NO5(M-H)-,346.0908；实测值346.0911。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物23，制得化合物24；化合物10替换为化合物24，制得白色固体OX-8(0.11g，收率25.6％，纯度98.88％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.88(s,1H),11.00(s,1H),8.65(t,J＝6.3Hz,1H),7.64(d,J＝7.7Hz,1H),7.41(s,1H),7.34(d,J＝7.8Hz,1H),7.27-7.15(m,2H),7.08-6.90(m,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.50-4.21(m,4H),3.81(s,2H),3.02-2.84(m,1H),2.60(d,J＝17.5Hz,1H),2.48-2.31(m,1H),2.07-1.93(m,1H)。
HRMS(ESI+):计算值C24H22F3N4O6(M+H)+,519.1486；实测值519.1482。
实施例9 化合物OX-9：(E)-N-((2-(2，6-二氧哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)-3-(4-(三氟甲基)苯基)丙酰胺的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为对三氟甲基苯甲醛，制得化合物25：(E)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)-3-(4-(三氟甲基)苯基)丙酸。三步收率共计45.3％。HRMS(ESI-):计算值C15H15F3NO4(M-H)-,330.0959；实测值330.0961。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物25，制得化合物26；化合物10替换为化合物26，制得黄色固体OX-9(0.12g，收率27.9％，纯度98.43％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.95(s,1H),11.05(s,1H),8.67(t,J＝6.3Hz,1H),7.64(d,J＝7.7Hz,1H),7.50-7.32(m,4H),7.25-7.13(m,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.51-4.22(m,4H),3.78(s,2H),2.99-2.83(m,1H),2.60(d,J＝17.5Hz,1H),2.47-2.32(m,1H),2.08-1.94(m,1H)。
HRMS(ESI+):计算值C24H22F3N4O5(M+H)+,503.1537；实测值503.1532。
实施例10 化合物OX-10：(E)-3-([1，1'-联苯]-4-基)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(羟基亚氨基)丙酰胺7的合成：
按照中间体-3的合成方法，将起始原料苯甲醛替换为对苯基苯甲醛，制得化合物27：(E)-3-([1，1'-联苯]-4-基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸。三步收率共计43.6％。HRMS(ESI-):计算值C20H20NO4(M-H)-,338.1398；实测值338.1390。
按照实施例1中化合物OX-1的合成方法，将起始原料中间体-3替换为化合物27，制得化合物28；化合物10替换为化合物28，制得淡黄色固体OX-10(0.10g，收率23.3％，纯度97.28％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.90(s,1H),11.05(s,1H),8.70(t,J＝6.3Hz,1H),7.80-7.65(m,4H),7.55-7.38(m,4H),7.33(s,1H)7.25-7.06(m,3H),5.11(dd,J＝13.3,5.0Hz,1H),4.56-4.23(m,4H),3.83(s,2H),3.00-2.83(m,1H),2.60(d,J＝17.7Hz,1H),2.48-2.32(m,1H),2.07-1.95(m,1H)。HRMS(ESI+):计算值C29H27N4O5(M+H)+,511.1976；实测值511.1972。
实施例11 化合物OX-11：(E)-3-(3-氯-4-甲基苯基)-N-(2-(2，6-二氧哌啶-3-基)-1，3-二氧异吲哚啉-5-基)-2-(羟基亚氨基)丙酰胺的合成：
步骤1：将(E)-3-(3-氯-4-甲基苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸(化合物21，1.25g，4.03mmol)溶解于10mL的N,N-二甲基甲酰胺中，依次加入EDCI(1.05g，5.49mmol)、HOBT(0.74g，5.49mmol)，加毕，室温搅拌1小时。加入5-氨基-2-(2,6-二氧哌啶-3-基)异吲哚啉-1,3-二酮(1.0g，3.66mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入50mL水中，降温析出黄色固体，抽滤，滤饼干燥，将残余物使用硅胶色谱柱(二氯甲烷：甲醇＝60：1，v/v)纯化，得到黄色固体29：(E)-3-(3-氯-4-甲基苯基)-N-(2-(2,6-二氧代哌啶-3-基)-1,3-二氧代异吲哚啉-5-基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酰胺(1.07g，收率50.2％)。
步骤2：化合物29(0.20g，0.35mmol)溶于10mL的盐酸-二氧六环溶液(4M)中，室温搅拌20小时。经TLC监测反应完全，反应液真空浓缩并干燥，将残余物使用硅胶色谱柱(二氯甲烷：甲醇＝60：1，v/v)纯化，得到黄白色固体OX-11(0.13g，收率75.0％，纯度97.77％)。1H NMR(300MHz,DMSO-d6)δ(ppm)12.40(s,1H),11.13(s,1H),10.64(s,1H),8.34(s,1H),8.20-8.11(m,1H),7.89(d,J＝8.2Hz,1H),7.32-7.22(m,2H),7.14(d,J＝7.8Hz,1H),5.13(dd,J＝12.9,5.3Hz,1H),3.90(s,2H),2.98-2.80(m,1H),2.66-2.53(m,2H),2.27(s,3H),2.11-2.00(m,1H)。HRMS(ESI+):计算值C23H20ClN4O6(M+H)+,483.1066；实测值483.1060。
实施例12 化合物OX-12：(E)-N-(2-(2,6-二氧哌啶-3-基)-1,3-二氧异吲哚啉-5-基)-3-(4-氟苯基)-2-(羟基亚氨基)丙酰胺的合成：
按照实施例11中化合物OX-11的合成方法，将起始原料化合物21替换为化合物11：(E)-3-(4-氟苯基)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)丙酸，制得化合物30；将化合物29替换为化合物30，制 得浅黄色固体OX-12(0.10g，收率58.8％，纯度96.95％)。1H NMR(300MHz,DMSO-d6)δ(ppm)12.05(s,1H),11.03(s,1H),10.58(s,1H),8.30(s,1H),8.20-8.09(m,1H),7.95(d,J＝8.2Hz,1H),7.40-7.28(m,1H),7.24-7.08(m,3H),5.13(dd,J＝12.9,5.3Hz,1H),3.88(s,2H),2.98-2.82(m,1H),2.59(d,J＝17.5Hz,1H),2.48-2.32(m,1H),2.09-1.94(m,1H)。HRMS(ESI+):计算值C22H18FN4O6(M+H)+,453.1205；实测值453.1200。
实施例13 化合物OX-13：(E)-N-(2-(2，6-二氧哌啶-3-基)-1，3-二氧异吲哚啉-5-基)-2-(羟基亚氨基)-3-(4-(三氟甲基)苯基)丙酰胺的合成：
按照实施例11中化合物OX-11的合成方法，将起始原料化合物21替换为化合物25：(E)-2-(((四氢-2H-吡喃-2-基)氧基)亚氨基)-3-(4-(三氟甲基)苯基)丙酸，制得化合物31；将化合物29替换为化合物31，制得浅黄色固体OX-13(0.09g，收率52.9％，纯度97.29％)。1H NMR(300MHz,DMSO-d6)δ(ppm)12.01(s,1H),10.98(s,1H),10.44(s,1H),8.33(s,1H),8.15-8.04(m,1H),7.95(d,J＝8.2Hz,1H),7.48-7.32(m,1H),7.23-7.08(m,3H),5.11(dd,J＝12.9,5.3Hz,1H),3.90(s,2H),3.00-2.82(m,1H),2.60(d,J＝17.7Hz,1H),2.48-2.32(m,1H),2.10-1.96(m,1H)。HRMS(ESI+):计算值C23H18F3N4O6(M+H)+,503.1173；实测值503.1168。
实施例14 化合物NX-1：(E)-N-((2-(2，6-二氧哌啶-3-基)-1，3-二氧异吲哚啉-5-基)甲基)-2-肼亚基-3-苯基丙酰胺的合成：
步骤1：将2-氧代-3-苯基丙酸(中间体-2，1.06g，6.47mmol)溶解于10mL的N,N-二甲基甲酰胺中，依次加入EDCI(1.24g，6.47mmol)、HOBT(0.87g，6.47mmol)，加毕，室温搅拌1小时。加入3-(5-氨甲基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮盐酸盐(1.00g，3.23mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入50mL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＝65：1，v/v)纯化，得到棕黄色固体32：N-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)甲基)-2-氧代-3-苯基丙酰胺(0.83g，收率59.0％)。
步骤2：化合物32(0.50g，1.15mmol)溶于10mL的乙酸溶液中(50％)，加入80％的水合肼(0.07g，1.15mmol)，室温搅拌5h，经TLC监测反应完全，将反应液倒入50mL饱和碳酸氢钠溶液中，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＝70：1，v/v)纯化，得淡黄色固体NX-1(0.15g，收率30.0％，纯度98.37％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.49(t,J＝6.4Hz,1H),7.82(d,J＝7.8Hz,1H),7.51(s,1H),7.38(d,J＝9.1Hz,1H),7.30(s,3H),7.21(s,2H),6.62(s,2H),5.13(dd,J＝13.3,5.1Hz,1H),4.50-4.25(m,4H),3.81(s,2H),2.99-2.85(m,1H),2.60(d,J＝17.0Hz,1H),2.47-2.33(m,1H),2.08-1.94(m,1H)。HRMS(ESI+):计算值C23H24N5O4(M+H)+,434.1823；实测值434.1816。
实施例15 化合物NX-2：N-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)甲基)-3-(4-氟苯基)-2-氧代丙酰胺的合成：
按照中间体-2的合成方法，将起始原料苯甲醛替换为对氟苯甲醛，制得化合物33：3-(4-氟苯基)-2-氧代丙酸。两步收率共计68.5％。HRMS(ESI-):计算值C9H6FO3(M-H)-,181.0306；实测值181.0306。
按照实施例14中化合物NX-1的合成方法，将起始原料中间体-2替换为化合物33，制得化合物34；将化合物32替换为化合物34，制得黄色固体NX-2(0.13g，收率26.0％，纯度99.60％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.39(t,J＝6.4Hz,1H),7.75(d,J＝7.8Hz,1H),7.50(s,1H),7.38-7.22(m,3H),7.15-7.04(m,2H),6.49(s,2H),5.13(dd,J＝13.3,5.1Hz,1H),4.50-4.23(m,4H),3.75(s,2H),3.00-2.83(m,1H),2.60(d,J＝17.3Hz,1H),2.47-2.32(m,1H),2.08-1.94(m,1H)。HRMS(ESI+):计算值C23H23FN5O4(M+H)+,452.1729；实测值452.1726。
实施例16 化合物NX-3：N-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)甲基)-2-氧代-3- (对甲苯基)丙酰胺的合成：
按照中间体-2的合成方法，将起始原料苯甲醛替换为对甲基苯甲醛，制得化合物35：2-氧代-3-(对甲苯基)丙酸。两步收率共计65.3％。HRMS(ESI-):计算值C10H9O3(M-H)-,177.0557；实测值177.0568。
按照实施例14中化合物NX-1的合成方法，将起始原料中间体-2替换为化合物35，制得化合物36；将化合物32替换为化合物36，制得棕黄色固体NX-3(0.17g，收率34.0％，纯度97.49％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.05(s,1H),8.45(t,J＝6.4Hz,1H),7.75(d,J＝7.8Hz,1H),7.50(s,1H),7.34-7.20(m,3H),7.12-7.02(m,2H),6.45(s,2H),5.11(dd,J＝13.3,5.1Hz,1H),4.50-4.23(m,4H),3.75(s,2H),3.00-2.85(m,1H),2.60(d,J＝17.5Hz,1H),2.47-2.32(m,1H),2.20(s,3H),2.07-1.92(m,1H)。HRMS(ESI+):计算值C24H26N5O4(M+H)+,448.1980；实测值448.1976。
实施例17 化合物NX-4：N-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)甲基)-2-氧代-3-(4-(三氟甲氧基)苯基)丙酰胺的合成：
按照中间体-2的合成方法，将起始原料苯甲醛替换为对三氟甲氧基苯甲醛，制得化合物37：2-氧代-3-(4-(三氟甲氧基)苯基)丙酸。两步收率共计62.3％。HRMS(ESI-):计算值C10H9O3(M-H)-,177.0557；实测值177.0568。
按照实施例14中化合物NX-1的合成方法，将起始原料中间体-2替换为化合物37，制得化合物38；将化合物32替换为化合物38，制得黄白色固体NX-4(0.11g，收率22.0％，纯度99.33％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.10(s,1H),8.56(t,J＝6.4Hz,1H),7.83(d,J＝7.8Hz,1H),7.45(s,1H),7.35-7.19(m,3H),6.99-6.87(m,2H),6.28(s,2H),5.11(dd,J＝13.3,5.1Hz,1H),4.46-4.20(m,4H),3.84(s,2H),3.00-2.83(m,1H),2.59(d,J＝17.0Hz,1H),2.47-2.30(m,1H),2.09-1.94(m,1H)。HRMS(ESI+):计算值C24H23F3N5O5(M+H)+,518.1646；实测值518.1642。
实施例18 化合物NX-5：3-(4-氯苯基)-N-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)甲基)-2-氧代丙酰胺的合成：
按照中间体-2的合成方法，将起始原料苯甲醛替换为对氯苯甲醛，制得化合物39：3-(4-氯苯基)-2-氧代丙酸。两步收率共计52.4％。HRMS(ESI-):计算值C9H6ClO3(M-H)-,197.0011；实测值197.0021。
按照实施例14中化合物NX-1的合成方法，将起始原料中间体-2替换为化合物39，制得化合物40；将化合物32替换为化合物40，制得黄白色固体NX-5(0.14g，收率28.0％，纯度97.90％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.95(s,1H),8.40(t,J＝6.4Hz,1H),7.88(d,J＝7.8Hz,1H),7.48(s,1H),7.40-7.29(m,3H),7.22(d,J＝7.8Hz,2H),6.33(s,2H),5.11(dd,J＝13.3,5.1Hz,1H),4.49-4.23(m,4H),3.80(s,2H),3.02-2.84(m,1H),2.60(d,J＝17.7Hz,1H),2.47-2.33(m,1H),2.07-1.93(m,1H)。HRMS(ESI+):计算值C23H23ClN5O4(M+H)+,468.1433；实测值468.1427。
实施例19 化合物NX-6：N-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)甲基)-2-氧代-3-(4-(三氟甲基)苯基)丙酰胺的合成：
按照中间体-2的合成方法，将起始原料苯甲醛替换为对三氟甲基苯甲醛，制得化合物41：2-氧代-3-(4-(三氟甲基)苯基)丙酸。两步收率共计53.6％。HRMS(ESI-):计算值C10H6F3O3(M-H)-,231.0275；实测值231.0268。
按照实施例14中化合物NX-1的合成方法，将起始原料中间体-2替换为化合物41，制得化合物42；将化合物32替换为化合物42，制得浅黄色固体NX-6(0.10g，收率20.0％，纯度97.51％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),8.49(t,J＝6.3Hz,1H),7.83(d,J＝7.8Hz,1H),7.55-7.43(m,3H),7.37-7.29(m,2H),7.19(d,J＝7.0Hz,1H),6.28(s,2H),5.11(dd,J＝13.3,5.1Hz,1H),4.51-4.25(m,4H),3.85(s,2H),2.98-2.85(m,1H),2.58(d,J＝17.0Hz,1H),2.47-2.32(m,1H),2.10-1.94(m,1H)。HRMS(ESI+):计算值 C24H23F3N5O4(M+H)+,502.1697；实测值502.1694。
实施例20 化合物NX-7：3-(3-氯-4-甲基苯基)-N-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)甲基)-2-氧代丙酰胺的合成：
按照中间体-2的合成方法，将起始原料苯甲醛替换为3-氯-4-甲基苯甲醛，制得化合物43：3-(3-氯-4-甲基苯基)-2-氧代丙酸。两步收率共计58.2％。HRMS(ESI-):计算值C10H8ClO3(M-H)-,211.0167；实测值211.0159。
按照实施例14中化合物NX-1的合成方法，将起始原料中间体-2替换为化合物43，制得化合物44；将化合物32替换为化合物44，制得浅黄白色固体NX-7(0.15g，收率30.0％，纯度97.31％)。1H NMR(400MHz,DMSO-d6)δ(ppm)11.02(s,1H),8.37(t,J＝6.4Hz,1H),7.66(d,J＝7.8Hz,1H),7.43(s,1H),7.40-7.32(m,3H),7.25(s,1H),7.23(d,J＝7.8Hz,1H),7.09(d,J＝9.5Hz,1H),5.11(dd,J＝13.3,5.1Hz,1H),4.51-4.21(m,4H),3.75(s,2H),2.96-2.85(m,1H),2.60(d,J＝17.3Hz,1H),2.45-2.31(m,1H),2.26(s,3H),2.03-1.94(m,1H)。HRMS(ESI+):计算值C24H25ClN5O4(M+H)+,482.1590；实测值482.1578。
实施例21 化合物CX-1：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)-3-苯基丙酰胺的合成：
将(E)-2-(甲氧基亚氨基)-3-苯基丙酸(中间体-4，0.31g，1.62mmol)溶解于6mL的N,N-二甲基甲酰胺中，依次加入EDCI(0.38g，2.02mmol)、HOBT(0.27g，2.02mmol)，加毕，室温搅拌1小时，加入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-1，0.25g，0.81mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入30mL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＝60：1，v/v)纯化，得到灰白色固体CX-1(0.08g，22.2％，纯度97.85％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.05(s,1H),9.08(t,J＝6.1Hz,1H),7.86(d,J＝7.8Hz,1H),7.52(s,1H),7.35-7.10(m,6H),5.11(dd,J＝13.3,5.0Hz,1H),4.50-4.23(m,4H),3.98(s,3H),3.85(s,2H),3.00-2.83(m,1H),2.60(d,J＝17.3Hz,1H),2.46-2.34(m,1H),2.08-1.95(m,1H)。HRMS(ESI+):计算值C24H25N4O5(M+H)+,449.1820；实测值449.1816。
实施例22 化合物CX-2：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)-3-(对甲苯基)丙酰胺的合成：
按照中间体-2、中间体-4的合成方法，将起始原料苯甲醛替换为对甲基苯甲醛，制得化合物45：(E)-2-(甲氧基亚氨基)-3-(对甲苯基)丙酸。三步收率共计62.5％。HRMS(ESI-):计算值C11H12NO3(M-H)-,206.0823；实测值206.0835。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物45，制得灰白色固体CX-2(0.11g，收率29.7％，纯度98.34％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.99(t,J＝6.1Hz,1H),7.95(d,J＝7.8Hz,1H),7.52(s,1H),7.35(d,J＝7.7Hz,2H),7.20(d,J＝7.8Hz,1H),7.05(d,J＝7.5Hz,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.52-4.25(m,4H),3.95(s,3H),3.82(s,2H),2.99-2.81(m,1H),2.60(d,J＝17.7Hz,1H),2.47-2.31(m,1H),2.23(s,3H),2.08-1.92(m,1H)。HRMS(ESI+):计算值C25H27N4O5(M+H)+,463.1976；实测值463.1972。
实施例23 化合物CX-3：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-3-(4-氟苯基)-2-(甲氧基亚氨基)丙酰胺的合成：
按照中间体-2、中间体-4的合成方法，将起始原料苯甲醛替换为对氟苯甲醛，制得化合物46：(E)-3-(4-氟苯基)-2-(甲氧基亚氨基)丙酸。三步收率共计65.3％。HRMS(ESI-):计算值C10H9FNO3(M-H)-, 210.0572；实测值210.0568。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物46，制得白色固体CX-3(0.13g，收率34.4％，纯度99.19％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.99(s,1H),8.89(t,J＝6.1Hz,1H),7.92(d,J＝7.6Hz,1H),7.55(s,1H),7.35-7.25(m,3H),7.05(t,J＝8.9Hz,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.50-4.25(m,4H),3.97(s,3H),3.78(s,2H),3.00-2.83(m,1H),2.60(d,J＝17.3Hz,1H),2.47-2.32(m,1H),2.08-1.95(m,1H)。HRMS(ESI+):计算值C24H24FN4O5(M+H)+,467.1725；实测值467.1720。
实施例24 化合物CX-4：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-3-(3-氟苯基)-2-(甲氧基亚氨基)丙酰胺的合成：
按照中间体-2、中间体-4的合成方法，将起始原料苯甲醛替换为3-氟苯甲醛，制得化合物47：(E)-3-(3-氟苯基)-2-(甲氧基亚氨基)丙酸。三步收率共计55.7％。HRMS(ESI-):计算值C10H9FNO3(M-H)-,210.0572；实测值210.0563。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物47，制得灰白色固体CX-4(0.10g，收率26.5％，纯度98.62％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.05(s,1H),8.86(t,J＝6.1Hz,1H),7.88(d,J＝7.6Hz,1H),7.48(s,1H),7.35-7.26(m,2H),7.15(d,J＝7.5Hz,2H),7.05-6.94(m,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.45-4.21(m,4H),3.95(s,3H),3.82(s,2H),3.02-2.85(m,1H),2.60(d,J＝17.3Hz,1H),2.47-2.32(m,1H),2.10-1.95(m,1H)。HRMS(ESI+):计算值C24H24FN4O5(M+H)+,467.1725；实测值467.1718。
实施例25 化合物CX-5：(E)-3-(4-氯苯基)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)丙酰胺的合成：
按照中间体-2、中间体-4的合成方法，将起始原料苯甲醛替换为对氯苯甲醛，制得化合物48：(E)-3-(4-氯苯基)-2-(甲氧基亚氨基)丙酸。三步收率共计59.6％。HRMS(ESI-):计算值C10H9ClNO3(M-H)-,226.0276；实测值226.0268。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物48，制得白色固体CX-5(0.10g，收率25.6％)。1H NMR(300MHz,DMSO-d6)δ(ppm)10.90(s,1H),9.05(t,J＝6.1Hz,1H),7.76(d,J＝7.6Hz,1H),7.50(s,1H),7.36-7.28(m,3H),7.16(t,J＝8.9Hz,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.52-4.28(m,4H),3.95(s,3H),3.79(s,2H),2.97-2.82(m,1H),2.60(d,J＝17.0Hz,1H),2.47-2.32(m,1H),2.08-1.96(m,1H)。HRMS(ESI+):计算值C24H24ClN4O5(M+H)+,483.1430；实测值483.1428。
实施例26 化合物CX-6：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)-3-(4-(三氟甲基)苯基)丙酰胺的合成：
按照中间体-2、中间体-4的合成方法，将起始原料苯甲醛替换为对三氟甲基苯甲醛，制得化合物49：(E)-2-(甲氧基亚氨基)-3-(4-(三氟甲基)苯基)丙酸。三步收率共计61.3％。HRMS(ESI-):计算值C11H9F3NO3(M-H)-,260.0540；实测值260.0537。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物49，制得灰色固体CX-6(0.09g，收率21.4％，纯度98.42％)。1H NMR(300MHz,DMSO-d6)δ(pp m)11.00(s,1H),8.94(t,J＝6.1Hz,1H),7.80(d,J＝7.6Hz,1H),7.52-7.44(m,3H),7.33-7.21(m,3H),5.11(dd,J＝13.3,5.0Hz,1H),4.50-4.25(m,4H),3.99(s,3H),3.75(s,2H),2.98-2.83(m,1H),2.61(d,J＝17.3Hz,1H),2.48-2.33(m,1H),2.10-1.96(m,1H)。HRMS(ESI+):计算值C25H24F3N4O5(M+H)+,517.1694；实测值517.1691。
实施例27 化合物CX-7：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)-3-(4-(三氟甲氧基)苯基)丙酰胺的合成：
按照中间体-2、中间体-4的合成方法，将起始原料苯甲醛替换为对三氟甲氧基苯甲醛，制得化合物50：(E)-2-(甲氧基亚氨基)-3-(4-(三氟甲氧基)苯基)丙酸。三步收率共计62.5％。HRMS(ESI-):计算值C11H9F3NO4(M-H)-,276.0489；实测值276.0475。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物50，制得灰白色固体CX-7(0.08g，收率18.6％，纯度98.66％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.94(t,J＝6.1Hz,1H),7.80(d,J＝7.6Hz,1H),7.49(s,1H),7.34(d,J＝7.2Hz,1H),7.21(d,J＝7.0Hz,2H),6.90(d,J＝7.5Hz,2H),5.11(dd,J＝13.3,5.0Hz,1H),4.46-4.24(m,4H),3.98(s,3H),3.78(s,2H),3.01-2.83(m,1H),2.60(d,J＝17.3Hz,1H),2.47-2.32(m,1H),2.07-1.96(m,1H)。HRMS(ESI+):计算值C25H24F3N4O6(M+H)+,533.1643；实测值533.1636。
实施例28 化合物CX-8：(E)-3-(3-氯-4-甲基苯基)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)丙酰胺的合成：
按照中间体-2、中间体-4的合成方法，将起始原料苯甲醛替换为3-氯-4-甲基苯甲醛，制得化合物51：(E)-3-(3-氯-4-甲基苯基)-2-(甲氧基亚氨基)丙酸。三步收率共计65.6％。HRMS(ESI-):计算值C11H11ClNO3(M-H)-,240.0433；实测值240.0438。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物51，制得白色固体CX-8(0.13g，收率32.5％，纯度99.92％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.00(s,1H),8.86(d,J＝6.6Hz,1H),7.65(d,J＝7.8Hz,1H),7.40(s,1H),7.35(d,J＝7.7Hz,1H),7.25(d,J＝7.8Hz,1H),7.19(s,1H),7.05(d,J＝7.9Hz,1H),5.11(dd,J＝13.3,5.0Hz,1H),4.52-4.21(m,4H),3.98(s,3H),3.80(s,2H),3.00-2.81(m,1H),2.61(d,J＝15.3Hz,1H),2.45-2.34(m,1H),2.27(s,3H),2.08-1.92(m,1H)。HRMS(ESI+):计算值C25H26ClN4O5(M+H)+,497.1586；实测值497.1582。
实施例29 化合物CX-9：(E)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(4-氟苯基)-2-(甲氧基亚氨基)乙酰胺的合成：
按照中间体-4的合成方法，将起始原料中间体-2替换为2-(4-氟苯基)-2-氧代乙酸，制得化合物52：(Z)-2-(4-氟苯基)-2-(甲氧基亚氨基)乙酸，收率78.3％。HRMS(ESI-):计算值C9H7FNO3(M-H)-,196.0415；实测值196.0425。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物52，制得黄白色固体CX-9(0.09g，收率25.0％，纯度96.88％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),9.34(t,J＝6.1Hz,1H),7.73(d,J＝7.9Hz,1H),7.65-7.54(m,3H),7.50(d,J＝7.9Hz,1H),7.30(t,J＝8.9Hz,2H),5.13(dd,J＝13.2,5.1Hz,1H),4.63-4.26(m,4H),3.96(s,3H),3.00-2.85(m,1H),2.61(d,J＝17.4Hz,1H),2.45-2.33(m,1H),2.07-1.97(m,1H)。HRMS(ESI+):计算值C23H22FN4O5(M+H)+,453.1569；实测值453.1568。
实施例30 化合物CX-10：(E)-2-(4-氯苯基)-N-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)乙酰胺的合成：
按照中间体-4的合成方法，将起始原料中间体-2替换为2-(4-氟苯基)-2-氧代乙酸，制得化合物53：(Z)-2-(4-氯苯基)-2-(甲氧基亚氨基)乙酸，收率75.6％。HRMS(ESI-):计算值C9H7ClNO3(M-H)-,212.0120；实测值212.0131。
按照实施例21中化合物CX-1的合成方法，将起始原料中间体-4替换为化合物53，制得浅黄色固体CX-10(0.10g，收率26.3％，纯度98.12％)。1H NMR(300MHz,DMSO-d6)δ(ppm)11.05(s,1H),9.25(t,J＝ 6.1Hz,1H),7.79(d,J＝7.9Hz,1H),7.66-7.54(m,3H),7.48(d,J＝7.9Hz,1H),7.25(t,J＝8.9Hz,2H),5.11(dd,J＝13.2,5.1Hz,1H),4.55-4.26(m,4H),3.90(s,3H),2.98-2.82(m,1H),2.60(d,J＝17.3Hz,1H),2.47-2.33(m,1H),2.08-1.99(m,1H)。HRMS(ESI+):计算值C23H22ClN4O5(M+H)+,469.1273；实测值469.1269。
实施例31 化合物CX-11：(E)-2-(2-氨基噻唑-4-基)-N-((2-(2，6-二氧嘧啶-3-基)-1-氧代异吲哚啉-5-基)甲基)-2-(甲氧基亚氨基)乙酰胺盐酸盐的合成：
步骤1：将(E)-2-(2-氨基噻唑-4-基)-2-(甲氧基亚氨基)乙酸(化合物54，0.5g，2.48mmol)溶于4mL的N,N-二甲基甲酰胺和10mL的四氢呋喃的混合溶剂中，冰浴下滴加4mL的二(三甲基硅基)氨基钠的四氢呋喃溶液(2M)，滴毕，冰浴条件下搅拌1h，0℃条件下滴加二碳酸二叔丁酯(0.58g，2.68mmol)，滴毕，室温条件下搅拌2h。经TLC监测反应完全，将反应液真空浓缩，残余物倒入20mL水中，用稀盐酸(3M)调节PH值至3-4，析出黄白色固体，静置使其完全析出，抽滤，滤饼干燥得浅黄色固体55：(E)-2-(2-((叔丁氧基羰基)氨基)噻唑-4-基)-2-(甲氧基亚氨基)乙酸(0.26g，收率34.6％)。
步骤2：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-1，0.15g，0.49mmol)溶解于6mL的N,N-二甲基甲酰胺中，依次加入化合物55(0.15g，0.49mmol)、HATU(0.28g，0.74mmol)，加毕，室温搅拌1小时。加入DIPEA(0.19g，1.47mmol)，室温过夜搅拌。经TLC监测反应完全，将反应液倒入30mL冰水中，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)纯化得棕黄色固体56：(E)-(4-(2-((2-(2，6-二氧代哌啶-3-基)-1-氧代异吲哚啉-5-基)甲基)氨基)-1-(甲氧基亚氨基)-2-氧代乙基)噻唑-2-基)氨基甲酸叔丁酯(0.11g，收率40.7％)。
步骤3：将化合物56(0.11g，0.19mmol)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CX-11(0.04g，收率45.9％，纯度98.87％)。1HNMR(300MHz,DMSO-d6)δ(ppm)11.01(s,1H),9.08(t,J＝6.1Hz,1H),7.95(s,1H),7.74(d,J＝7.8Hz,1H),7.56(s,1H),7.49(d,J＝7.9Hz,1H),7.26(s,2H),5.13(dd,J＝13.3,5.0Hz,1H),4.53-4.28(m,4H),3.99(s,3H),3.01-2.85(m,1H),2.60(d,J＝19.1Hz,1H),2.45-2.30(m,1H),2.07-1.96(m,1H)。HRMS(ESI+):计算值C20H21N6O5S(M+H)+,457.1289；实测值457.1278。
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。
对照组为空白溶剂组。
以下是本测试例所用抗体：
一抗：
Anti-GSPT1：Abcam ab234433
Anti-IKZF1：Cell signaling technology(CST)5443S
Anti-IKZF3：Cell signaling technology(CST)15103S
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，本发明各系列化合物中的优势化合物的降解效果优于临床药物，其他大部分化合物都与临床药物降解效果相持。可见，肟类化合物OX、NX、CX系列对GSPT1蛋白的降解效果都能够有明显提高。
本发明以化合物OX-7为例，提供了其对AML细胞中的GSPT1降解作用呈现浓度依赖和时间依赖的图谱，见图1、图2。其中，图1A为化合物OX-7与临床药物CC-90009在KG-1细胞中对GSPT1的降解作用。图1B为化合物OX-7与临床药物CC-90009在MOLM-13细胞中对GSPT1的降解作用。
从图中可以看出，本发明化合物OX-7对KG-1、MOLM-13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：本发明化合物抗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
从表中各系列化合物的抗增殖活性可以看出，本发明化合物OX、NX、CX系列对KG-1细胞的抗增殖抑制活性明显提高，优于临床药物CC-90009。其中，各系列优势化合物如OX-7、NX-7、CX-7、CX-8抗增 殖抑制活性优异，IC50处于1-10nM。
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，本发明各系列化合物中优势化合物的抗增殖抑制活性优于CC-90009，其他化合物也与CC-90009相持。因此，可见本发明的肟类化合物OX、NX、CX系列对多种AML细胞的抗增殖抑制活性显著提高。
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 (10)

  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：
    X选自-OH、-NH2、-OCH3或-NHCOCH3；
    环Y选自C6-C12芳基、5-10元杂芳基、C3-C8环烷基或3-10元杂环基，所述芳基、杂芳基、环烷基和杂环基未被取代或任选地被1-3个Ra取代；
    R1选自
    R2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、卤素或C1-C6烷基；
    R5选自氢、氘、C1-C12烷基、
    Z选自-CH2-，-CD2-，-C(O)-或-C(S)-；
    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；
    每个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；
    每个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、p分别独立地选自0、1、2或3。
  2. 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，X、Y、Z、m、n、p如通式I中所定义。
  3. 根据权利要求1或2所述的化合物，其特征在于：
    X选自-OH、-NH2或-OCH3。
  4. 根据权利要求1所述的化合物，其特征在于：
    R2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；
    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；
    环Y选自C6-C12芳基或5-10元杂芳基。
  5. 根据权利要求4所述的化合物，其特征在于：
    R3选自氢、C1-C6烷基、卤代C1-C6烷基、C2-C6烯基、C2-C6炔基、卤素、氰基或硝基；
    R4选自氢、氘、卤素或C1-C6烷基；
    R5选自氢、氘或C1-C12烷基；
    m、n分别独立地选自1；
    p选自0或1。
  6. 根据权利要求1所述的化合物，其特征在于选自：

  7. 一种药物组合物，其特征在于，包含治疗有效量的权利要求1-5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。
  8. 权利要求1-6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。
  9. 根据权利要求8所述的用途，其特征在于，所述相关疾病为癌症、病毒感染、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。
  10. 权利要求1-6任一项所述的化合物在制备GSPT1降解剂中的用途。
PCT/CN2024/103853 2023-06-06 2024-07-05 一种含肟结构的gspt1蛋白降解剂及其制备方法和应用 Ceased WO2024251301A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN202310660635.3 2023-06-06
CN202310660635.3A CN119080738A (zh) 2023-06-06 2023-06-06 一种含肟结构的gspt1蛋白降解剂及其制备方法和应用

Publications (1)

Publication Number Publication Date
WO2024251301A1 true WO2024251301A1 (zh) 2024-12-12

Family

ID=93661049

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2024/103853 Ceased WO2024251301A1 (zh) 2023-06-06 2024-07-05 一种含肟结构的gspt1蛋白降解剂及其制备方法和应用

Country Status (2)

Country Link
CN (1) CN119080738A (zh)
WO (1) WO2024251301A1 (zh)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022200857A1 (en) * 2021-03-22 2022-09-29 Monte Rosa Therapeutics Ag Pharmaceutical compositions for use in the prevention and treatment of a disease or disorder caused by or associated with one or more premature termination codons
CN115304606A (zh) * 2021-06-21 2022-11-08 清华大学 一种同时靶向btk和gspt1蛋白的降解剂
WO2023070120A1 (en) * 2021-10-22 2023-04-27 Biotheryx, Inc. Ketoamides for treating malignancy

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2022200857A1 (en) * 2021-03-22 2022-09-29 Monte Rosa Therapeutics Ag Pharmaceutical compositions for use in the prevention and treatment of a disease or disorder caused by or associated with one or more premature termination codons
CN115304606A (zh) * 2021-06-21 2022-11-08 清华大学 一种同时靶向btk和gspt1蛋白的降解剂
WO2023070120A1 (en) * 2021-10-22 2023-04-27 Biotheryx, Inc. Ketoamides for treating malignancy

Also Published As

Publication number Publication date
CN119080738A (zh) 2024-12-06

Similar Documents

Publication Publication Date Title
KR102858043B1 (ko) Il-12, il-23 및/또는 ifn-알파의 조정과 관련된 상태의 치료를 위한 아미드-치환된 헤테로시클릭 화합물
KR101866858B1 (ko) Lsd1의 아릴사이클로프로필아민 기반 디메틸라아제 억제제 및 이의 의학적 이용
US9453048B2 (en) IAP antagonists
CN104903301B (zh) 可用于调节IL‑12、IL‑23和/或IFNα的烷基酰胺取代的嘧啶化合物
KR20180081584A (ko) Il-12, il-23 및/또는 ifn 알파 반응의 조정제로서 유용한 이미다조피리다진 화합물
KR102602558B1 (ko) Il-12, il-23 및/또는 ifn 알파 반응의 조정제로서의 포스핀 옥시드 알킬 아미드 치환된 헤테로아릴 화합물
WO2008054454A2 (en) Nitrogen-containing heterocycle derivatives, pharmaceutical compositions, and methods of use thereof as antiviral agents
JPH09208584A (ja) アミド誘導体、およびそれを含有する医薬製剤、および合成中間体
JP2016511237A (ja) 選択的ｈｄａｃ３阻害剤
AU2022272725A1 (en) Substituted heterocyclic compounds
US20230150979A1 (en) Amide-disubstituted pyridine or pyridazine compounds
KR20240008337A (ko) 치환된 헤테로시클릭 화합물
WO2022241175A1 (en) Substituted heterocyclic compounds
CN105189469B (zh) 2-氨基-3,4-二氢喹唑啉衍生物及其作为组织蛋白酶d抑制剂的用途
CA2700113A1 (en) Indazole acrylic acid amide compound
EP3628664A1 (en) Irreversible inhibitors of kras g12c mutant
WO2024251301A1 (zh) 一种含肟结构的gspt1蛋白降解剂及其制备方法和应用
WO2024222918A1 (zh) 一种苯并六元杂环类gspt1蛋白降解剂及其应用
CN105228609A (zh) 新的转录因子调节剂
WO2025241483A1 (zh) 一种n-(4-甲基苯基)-吲哚类化合物、其药物组合物及应用
WO2009038411A2 (en) Beta-secretase inhibiting compounds having oxo-dihydro-pyrazole moiety
WO2017008681A1 (zh) 酰胺类衍生物、其制备方法及其在医药上的用途
CN117279906A (zh) 经取代的杂环化合物
CN115838354A (zh) 一种甲状腺激素受体β激动剂化合物及其制备方法和应用
KR20210080378A (ko) 비만의 치료

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: 24818826

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE