WO2022007659A1 - 杂环类免疫调节剂 - Google Patents

杂环类免疫调节剂 Download PDF

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WO2022007659A1
WO2022007659A1 PCT/CN2021/102873 CN2021102873W WO2022007659A1 WO 2022007659 A1 WO2022007659 A1 WO 2022007659A1 CN 2021102873 W CN2021102873 W CN 2021102873W WO 2022007659 A1 WO2022007659 A1 WO 2022007659A1
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alkyl
mmol
compound
cycloalkyl
pharmaceutically acceptable
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French (fr)
Inventor
陈向阳
庞育成
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Beijing Innocare Pharma Tech Co Ltd
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Beijing Innocare Pharma Tech Co Ltd
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Priority to MX2023000382A priority Critical patent/MX2023000382A/es
Priority to CA3189041A priority patent/CA3189041A1/en
Priority to KR1020237004029A priority patent/KR20230037035A/ko
Priority to EP21838438.6A priority patent/EP4177247A4/en
Priority to AU2021303635A priority patent/AU2021303635A1/en
Priority to US18/014,623 priority patent/US20230271933A1/en
Priority to CN202180048474.1A priority patent/CN115776983A/zh
Priority to JP2023500425A priority patent/JP7712703B2/ja
Publication of WO2022007659A1 publication Critical patent/WO2022007659A1/zh
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    • 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
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/496—Non-condensed piperazines containing further heterocyclic rings, e.g. rifampin, thiothixene or sparfloxacin
    • 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
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
    • 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
    • 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
    • A61K31/5386—1,4-Oxazines, e.g. morpholine spiro-condensed or forming part of bridged ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • 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
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • 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
    • 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
    • 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
    • C07D401/14—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 three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/08—Bridged systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D498/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D498/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and oxygen atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D498/08—Bridged systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/07—Optical isomers

Definitions

  • the present invention relates to heterocyclic compounds or pharmaceutically acceptable salts thereof as immunomodulators.
  • the present invention also relates to a process for the preparation of said compound or a pharmaceutically acceptable salt thereof.
  • the present invention further relates to uses and methods of using the compounds or pharmaceutically acceptable salts thereof in immunomodulation, anticancer and anti-inflammatory.
  • Lenalidomide is an immunomodulatory drug (IMiD) with various mechanisms of action.
  • IMD immunomodulatory drug
  • these immunomodulators regulate the function of the CRL4 CRBN -E3 ubiquitin ligase complex by binding to E3 ubiquitin ligase, resulting in Ikaros (IKZF1), Aiolos (IKZF3), protein kinase CK1 ⁇ (CK1 ⁇ ),
  • the transformation termination factor GSPT1 is ubiquitinated and degraded by the 26S proteasome, thereby changing the secretion of various cytokines (such as IL-2, IL-6, IL-10, TNF ⁇ , IL-1 ⁇ , etc.) and affecting the activity of immune cells.
  • cytokines such as IL-2, IL-6, IL-10, TNF ⁇ , IL-1 ⁇ , etc.
  • dommine compounds can inhibit the angiogenesis of tumor cells by inhibiting vascular endothelial growth factor (VEGF), and can also directly inhibit the proliferation of tumor cells and induce the decomposition of abnormal cells.
  • VEGF vascular endothelial growth factor
  • duramide compounds have received extensive attention in the treatment of many types of malignant tumors and immune diseases, such as multiple myeloma, myelodysplastic syndrome, hematological tumors/solid tumors (such as lymphoma, non-small cell lung cancer, pancreas, prostate, brain, kidney, ovary, etc.), and systemic lupus erythematosus.
  • duramide compounds can also be used in combination with other drugs to treat diseases, such as small molecule targeted drugs, chemotherapy drugs, macromolecular drugs (such as PD-1 antibody, CD20 antibody, CD19 antibody, etc.).
  • the present invention relates to a compound with a structure represented by general formula (I), which is used as an immunomodulator to effectively regulate the expression of IL-2, IL-6, IL-10, TNF ⁇ , VEGF and other proteins by combining with E3 ubiquitin ligase. expression and/or biological function.
  • general formula (I) which is used as an immunomodulator to effectively regulate the expression of IL-2, IL-6, IL-10, TNF ⁇ , VEGF and other proteins by combining with E3 ubiquitin ligase. expression and/or biological function.
  • C xy represents the range of carbon atoms, where x and y are integers, for example, C 3-8 cycloalkyl represents a cycloalkyl group with 3-8 carbon atoms, that is, with 3, 4, 5, 6, Cycloalkyl of 7 or 8 carbon atoms. It should also be understood that, “C 3-8” further includes any sub-range comprised therein, e.g. C 3-7, C 3- 6, C 4-7, C 4-6, C 5-6 and the like.
  • Alkyl refers to a saturated straight or branched chain hydrocarbyl group containing 1 to 20 carbon atoms, eg, 1 to 8 carbon atoms, 1 to 6 carbon atoms, or 1 to 4 carbon atoms.
  • alkyl groups 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, etc
  • Alkylene refers to a divalent group of a straight or branched chain saturated hydrocarbon containing 1 to 20 carbon atoms, eg, 1 to 6 carbon atoms or 1 to 4 carbon atoms.
  • alkylene groups include -CH 2 -, - CH (CH 3) -, - CH 2 CH 2 -, - CH 2 CH 2 CH 2 -, - (CH 3) C (CH 3) -, -CH 2 CH 2 CH 2 CH 2 -, - CH 2 CH (CH 3) CH 2 - and the like.
  • Cycloalkyl refers to a saturated cyclic hydrocarbyl substituent containing 3 to 14 carbon ring atoms. Cycloalkyl groups may be monocarbocyclic rings, typically containing 3 to 8, 3 to 7 or 3 to 6 carbon ring atoms. Non-limiting examples of monocyclic cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and the like. Cycloalkyl may also be bi- or tricyclic fused, bridged or spiro together, such as decalinyl, bicyclo[2.2.2]octane, spiro[3.3]heptane, and the like.
  • Heterocyclyl or heterocycle refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic group comprising 3 to 20 ring atoms, such as may be 3 to 14, 3 to 12, 3 to 10 , 3 to 8, 3 to 6, or 5 to 6 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O) m (wherein m is an integer from 0 to 2), but not including The ring moiety of -OO-, -OS- or -SS-, the remaining ring atoms being carbon.
  • It preferably includes 3 to 12 ring atoms, more preferably 3 to 10 ring atoms, more preferably 4 to 7 ring atoms, more preferably 4 to 6 ring atoms, most preferably 5 or 6 ring atoms, of which 1 to 4 is a heteroatom, more preferably 1 to 3 are heteroatoms, and most preferably 1 to 2 are heteroatoms.
  • Non-limiting examples of monocyclic heterocyclyl groups include pyrrolidinyl, oxetanyl, piperidinyl, piperazinyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydrothiopyranyl, morpholinyl, thiomorpho Linoyl, homopiperazinyl, azetidinyl, etc.
  • Polycyclic heterocyclic groups include fused, bridged or spiro polycyclic heterocyclic groups such as octahydrocyclopentadieno[c]pyrrole, octahydropyrrolo[1,2-a]pyrazine, 3,8-diaza Heterobicyclo[3.2.1]octane, 5-azaspiro[2.4]heptane, 2-oxa-7-azaspiro[3.5]nonane, etc.
  • Aryl or aromatic ring refers to an aromatic monocyclic or fused polycyclic group containing 6 to 14 carbon atoms, preferably 6 to 10 membered, such as phenyl and naphthyl, most preferably phenyl.
  • the aryl ring can be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein the ring linked to the parent structure is an aryl ring, non-limiting examples include:
  • Heteroaryl or heteroaromatic ring refers to a heteroaromatic system comprising 5 to 14 ring atoms, wherein 1 to 4 ring atoms are selected from heteroatoms including oxygen, sulfur and nitrogen.
  • Heteroaryl is preferably 5 to 10-membered, more preferably heteroaryl is 5- or 6-membered, such as furyl, thienyl, pyridyl, pyrrolyl, pyrimidinyl, pyrazinyl, pyrazolyl, imidazolyl, tetra oxazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl and the like.
  • the heteroaryl ring can be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring linked to the parent structure is a heteroaryl
  • Halogen refers to fluorine, chlorine, bromine or iodine.
  • Cyano refers to -CN.
  • Carbonyl refers to a -C(O)- group.
  • heterocyclic group optionally substituted with alkyl means that an alkyl group may, but need not, be present, and the expression includes both instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group .
  • Substituted means that one or more hydrogen atoms in a group, preferably 5, more preferably 1 to 3 hydrogen atoms, independently of each other, are substituted by the corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the person skilled in the art can determine (either experimentally or theoretically) possible or impossible substitutions without undue effort. For example, amino or hydroxyl groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (eg, olefinic) bonds.
  • the substituents include but are not limited to halogen, cyano, nitro, oxo, -SF 5 , C 1-4 alkyl, C 3-7 cycloalkyl, 4-7 membered heterocyclyl, phenyl, 5 -6-membered heteroaryl, etc.
  • “Isomers” refer to compounds that have the same molecular formula but differ in the nature or order in which their atoms are bonded or in the spatial arrangement of their atoms. Isomers that differ in the arrangement of atoms in space are called “stereoisomers”. Stereoisomers include optical isomers, geometric isomers and conformational isomers.
  • the compounds of the present invention may exist in the form of optical isomers.
  • Optical isomers include enantiomers and diastereomers.
  • Enantiomers are two stereoisomers that are mirror images of each other but are not mutually superimposable.
  • a racemic mixture or racemate refers to a mixture of equal numbers of left and right-handed enantiomers of a chiral molecule.
  • Diastereomer means that two stereoisomers are not mirror images of each other and are not superimposable.
  • optical isomer When the optical isomer is a single isomer and its absolute configuration is determined, it is the absolute configuration of "R” or “S” according to the configuration of the substituent on the chiral carbon atom; when the absolute configuration of the optical isomer is The configuration is not determined, it is (+) or (-) depending on the optical rotation value measured. Methods for the preparation and separation of optical isomers are known in the art.
  • the compounds of the present invention may also exist as geometric isomers.
  • the present invention contemplates various geometric isomers and mixtures thereof resulting from the distribution of substituents around carbon-carbon double bonds, carbon-nitrogen double bonds, cycloalkyl or heterocyclic groups. Substituents around carbon-carbon double bonds or carbon-nitrogen bonds are assigned the Z or E configuration, and substituents around cycloalkyl or heterocycles are assigned the cis or trans configuration.
  • the compounds of the present invention may also exhibit tautomerism, such as keto-enol tautomerism.
  • the present invention includes any tautomeric or stereoisomeric forms and mixtures thereof, and is not limited to any one tautomeric or stereoisomeric form used in the naming of the compounds or chemical structural formulae.
  • isotopes refer to all isotopes of atoms occurring in the compounds of the present invention. Isotopes include those atoms with the same atomic number but different mass numbers. Examples of isotopes suitable for incorporation into the compounds of the present invention are hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, and chlorine, such as, but not limited to, 2 H(D), 3 H, 13 C, 14 C, 15 N, respectively , 17 O, 18 O, 31 P, 32 P, 35 S, 18 F and 36 Cl.
  • Isotopically labeled compounds of the present invention can generally be prepared by conventional techniques known to those skilled in the art or by methods analogous to those described in the accompanying Examples, using the appropriate isotopically labeled reagent in place of the non-isotopically labeled reagent.
  • Such compounds have various potential uses, such as as standards and reagents in the determination of biological activity.
  • stable isotopes such compounds have the potential to advantageously alter biological, pharmacological or pharmacokinetic properties.
  • Deuterium (D) is a preferred isotope in the present invention, eg the hydrogen in methyl, methylene or methine may be replaced by deuterium.
  • prodrugs refers to a derivative that is converted to a biologically active compound of the present invention under physiological conditions in vivo, eg, by oxidation, reduction, hydrolysis, etc., each with or without enzymes.
  • prodrugs are compounds wherein the amino group in the compounds of the invention is acylated, alkylated or phosphorylated, such as eicosanoylamino, alanylamino, pivaloyloxymethylamino, or Where the hydroxyl group is acylated, alkylated, phosphorylated or converted to a borate, e.g.
  • “Pharmaceutically acceptable salts” or “pharmaceutically acceptable salts” refer to salts prepared from pharmaceutically acceptable bases or acids, including inorganic bases or acids and organic bases or acids. Where the compounds of the present invention contain one or more acidic or basic groups, the present invention also includes their corresponding pharmaceutically acceptable salts. Thus, the compounds of the invention which contain acidic groups can exist in salt form and can be used according to the invention, for example as alkali metal salts, alkaline earth metal salts or as ammonium salts. More precise examples of such salts include sodium, potassium, calcium, magnesium or salts with ammonia or organic amines such as ethylamine, ethanolamine, triethanolamine or amino acids.
  • the compounds according to the invention which contain basic groups can exist in salt form and can be used according to the invention in the form of their addition salts with inorganic or organic acids.
  • suitable acids include hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, nitric acid, methanesulfonic acid, p-toluenesulfonic acid, naphthalenedisulfonic acid, oxalic acid, acetic acid, tartaric acid, lactic acid, salicylic acid, benzoic acid, formic acid, propylene acid, pivalic acid, malonic acid, succinic acid, pimelic acid, fumaric acid, maleic acid, malic acid, sulfamic acid, phenylpropionic acid, gluconic acid, ascorbic acid, isonicotinic acid, citric acid, Adipic acid and other acids known to those skilled in the art.
  • the present invention includes, in addition to the salt forms mentioned, inner or betaine salts.
  • the respective salts can be obtained by conventional methods known to those skilled in the art, for example by contacting these with organic or inorganic acids or bases in solvents or dispersants or by anion exchange or cation exchange with other salts.
  • a “pharmaceutical composition” refers to a compound containing one or more of the compounds described herein, or pharmaceutically acceptable salts, stable isotope derivatives, isomers, prodrugs, and mixtures thereof, and other components such as pharmaceutically acceptable A combination of carriers and excipients.
  • the purpose of the pharmaceutical composition is to facilitate the administration to the organism, facilitate the absorption of the active ingredient and then exert the biological activity.
  • a compound of the present invention or “a compound of the present invention” in this application, it includes all such compound forms, such as pharmaceutically acceptable salts, stable isotope derivatives, isomers thereof body, prodrugs and mixtures thereof.
  • Carcer/tumor includes, but is not limited to, digestive/gastrointestinal cancer, colon cancer, liver cancer, skin cancer (including mast cell tumor and squamous cell carcinoma), breast cancer, ovarian cancer, prostate cancer, lymphoma, leukemia (including acute myeloid leukemia and chronic myeloid leukemia), kidney cancer, lung cancer, muscle cancer, bone cancer, bladder cancer, brain cancer, melanoma (including oral and metastatic melanoma), Kaposi's sarcoma (including Myeloma of multiple myeloma), myeloproliferative diseases, proliferative diabetic retinopathy, vascular proliferation-related disorders/tumors, etc.
  • leukemia including acute myeloid leukemia and chronic myeloid leukemia
  • kidney cancer including acute myeloid leukemia and chronic myeloid leukemia
  • kidney cancer including acute myeloid leukemia and chronic myeloid leukemia
  • kidney cancer including acute myeloid leukemia and chronic myeloid leukemia
  • “Inflammatory disease” includes, but is not limited to, arthritis, Hashimoto's thyroiditis, autoimmune hemolytic anemia, autoimmune atrophic gastritis of pernicious anemia, autoimmune encephalomyelitis, autoimmune orchitis, Goode Pascius disease, autoimmune thrombocytopenia, sympathetic ophthalmia, myasthenia gravis, Graves disease, primary biliary cirrhosis, hepatitis, primary sclerosing cholangitis, chronic invasive hepatitis, non- Alcoholic fatty liver disease, nonalcoholic steatohepatitis, ulcerative colitis, membranous glomerulopathy, systemic lupus erythematosus, rheumatoid arthritis, psoriatic arthritis, Sjögren's syndrome, Wright syndrome, Polymyositis, dermatomyositis, type I interferon disease including Aicardi-Goutines syndrome and other systemic s
  • a “therapeutically effective amount” is meant to include an amount of a compound of the invention effective to treat or prevent the disease.
  • Patient refers to mammals, especially humans.
  • the present invention relates to a compound represented by general formula (I), or a pharmaceutically acceptable salt, stable isotope derivative, isomer and prodrug thereof:
  • Ring A is a 4-10 membered heterocycle containing N;
  • R 1 is D or halogen
  • R 2 and R 3 are each independently selected from H, D, halogen, cyano, C 1-6 alkyl, C 3-6 cycloalkyl or 3-8 membered heterocyclyl, but both cannot be H and / or D, or R 2 and R 3 and the carbon atoms to which they are attached together form a C 3-6 cycloalkane or a 3-8 membered heterocyclic ring;
  • R 4 is D, halogen, cyano, oxo, C 1-6 alkyl, C 3-6 cycloalkyl, -OC 1-6 alkyl, -C(O)C 1-6 alkyl, -C (O)C 3-6 cycloalkyl, -S(O) 2 C 1-6 alkyl, -S(O) 2 C 3-6 cycloalkyl, aryl or heteroaryl, wherein the alkyl , cycloalkyl, aryl and heteroaryl optionally substituted with D, halogen, cyano, C 1-2 alkyl or fluoro C 1-2 alkyl at one or more hydrogens;
  • n is an integer 0-4;
  • n is an integer 0-2.
  • Ring A is a 4-10 membered monocyclic heterocycle containing one N atom, or a fused, bridged or spirobicyclic heterocycle (such as morpholine, piperidine, thiomorpholine-1,1- dioxide, 2-oxa-5-azabicyclo [2.2.1] heptane and the like); R 4 is H, D, halo, oxo, -CF 3 or C 1- 6 alkyl.
  • Ring A is a 6-10 membered monoheterocycle containing two N atoms, or a fused, bridged, or spirobicyclic heterocycle (eg, piperazine, 3,6-diazabicyclo[3.1.
  • R 4 is C 1-6 alkyl, C 3-6 cycloalkyl, -C ( O)C 1-6 alkyl, -C(O)C 3-6 cycloalkyl, -S(O) 2 C 1-6 alkyl, -S(O) 2 C 3-6 cycloalkyl, benzene or 5-6 membered heteroaryl containing N, O and/or S, wherein one or more hydrogens of the alkyl, cycloalkyl, phenyl and heteroaryl are optionally replaced by D, halogen, cyano , C 1-2 alkyl or fluoro C 1-2 alkyl substituted.
  • Ring A is piperazinyl;
  • C R 4 is a piperazine and the second N atom 1-6 alkyl, C 3-6 cycloalkyl, -C (O) C 1- 6 alkyl, -C(O)C 3-6 cycloalkyl, -S(O) 2 C 1-6 alkyl, phenyl, pyridyl or pyrimidinyl, wherein the alkyl, cycloalkyl, benzene
  • One or more hydrogens of the radical, pyridyl or pyrimidinyl are optionally substituted with D, halogen, cyano or C1-2 alkyl; n is 0 or 1.
  • R 2 is H;
  • R 3 is cyano, C 1-6 alkyl or C 3-6 cycloalkyl.
  • R 2 and R 3 are methyl.
  • R 2 and R 3 together form a C 3-6 cycloalkane with the carbon atom to which they are attached.
  • n is zero.
  • the compound of general formula (I) has the structure shown in the following general formula (II):
  • R 5 is H, C 1-6 alkyl, C 3-6 cycloalkyl, -C(O)C 1-6 alkyl, -C(O)C 3-6 cycloalkyl, -S(O) 2 C 1-6 alkyl, phenyl or 5-6 membered heteroaryl containing N, O and/or S, wherein one or more hydrogens of the alkyl, cycloalkyl, phenyl and heteroaryl optionally replaced by D, halogen, cyano or C 1-2 substituted alkyl.
  • R 5 is phenyl, pyridyl or pyrimidinyl, wherein said phenyl, optionally one or two hydrogen pyridinyl and pyrimidinyl is substituted with F or cyano.
  • the present invention further relates to the following compounds 1-22, or pharmaceutically acceptable salts, stable isotope derivatives, isomers, prodrugs and mixtures thereof:
  • the compounds of the present invention can effectively inhibit the proliferation of NCI-H929 cells, preferably with an IC 50 of less than 50 nM, more preferably with an IC 50 of less than 10 nM.
  • the compounds of the present invention have a significant inhibitory effect on the secretion of TNF ⁇ in human PBMC cells, preferably with an IC 50 of less than 20 nM, more preferably with an IC 50 of less than 10 nM.
  • Compounds of the invention also has a human PBMC cells secreting IL-2 stimulation significantly, preferably less than 50 nM EC 50, EC 50 and more preferably less than 10nM.
  • the present invention also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof, stable isotope derivatives, isomers, prodrugs and mixtures thereof, and a or multiple pharmaceutically acceptable carriers or excipients.
  • One aspect of the present invention provides a compound represented by general formula (I) or a pharmaceutically acceptable salt, stable isotope derivative, isomer, prodrug and mixture thereof, or the pharmaceutical composition for regulating or inhibiting E3
  • the activity of ubiquitin ligase thereby affecting the expression and/or biological function of proteins including but not limited to Aiolos, Ikaros, Helios, CK1 ⁇ , GSPT1, IL-2, IL-6, TNF ⁇ , IFN ⁇ , VEGF, etc.
  • Another aspect of the present invention provides a method for treating or preventing related diseases mediated by Aiolos, Ikaros, Helios, CK1 ⁇ , GSPT1, IL-2, IL-6, TNF ⁇ , IFN ⁇ , VEGF, etc.
  • diseases include but are not limited to hematological tumors (such as multiple myeloma, lymphoma, leukemia, etc.), solid tumors (such as lung cancer, prostate cancer, head and neck cancer, breast cancer, ovarian cancer, uterine cancer, pancreatic cancer, colon cancer, Rectal cancer, gastric cancer, esophageal cancer, brain cancer, liver cancer, kidney cancer, skin cancer, epithelial cancer, bladder cancer, neuroblastoma, etc.), autoimmune diseases (such as system
  • the present invention also relates to a pharmaceutical composition
  • a pharmaceutical composition comprising the compound represented by the general formula (I) or a pharmaceutically acceptable salt thereof, stable isotope derivatives, isomers, prodrugs and mixtures thereof and at least one An additional drug, wherein the at least one additional drug can be a small molecule chemotherapeutic agent (such as NSAIDs, steroid anti-inflammatory drugs, kinase targeting drugs, cytotoxic drugs, DNA damage related drugs, etc.) or macromolecular immune and/or inflammatory modulators (such as PD-1 antibody, CD20 antibody, CD19 antibody, TNF ⁇ antibody, IL-6 antibody, etc.).
  • a small molecule chemotherapeutic agent such as NSAIDs, steroid anti-inflammatory drugs, kinase targeting drugs, cytotoxic drugs, DNA damage related drugs, etc.
  • macromolecular immune and/or inflammatory modulators such as PD-1 antibody, CD20 antibody, CD19 antibody, TNF ⁇ antibody, IL-6 antibody, etc.
  • the drug can be in any pharmaceutical dosage form, including but not limited to tablets, capsules, solutions, freeze-dried preparations, and injections.
  • the pharmaceutical formulations of the present invention may be administered in dosage unit form containing a predetermined amount of active ingredient per dosage unit.
  • a unit may contain, for example, from 0.1 mg to 500 mg, preferably from 0.5 mg to 100 mg, of a compound of the invention, depending on the condition to be treated, the method of administration and the age, weight and condition of the patient.
  • pharmaceutical formulations of this type can be prepared using methods known in the pharmaceutical art, such as by admixing the active ingredient with one or more excipients and/or adjuvants.
  • compositions of the present invention may be suitable for administration by any desired suitable method, such as oral (including buccal or sublingual), rectal, nasal, topical (including buccal, sublingual or transdermal), vaginal or parenteral (including subcutaneous, intramuscular, intravenous or intradermal) methods of administration.
  • oral including buccal or sublingual
  • rectal including buccal or sublingual
  • nasal including buccal, sublingual or transdermal
  • vaginal or parenteral including subcutaneous, intramuscular, intravenous or intradermal
  • the present invention also provides methods for preparing the compounds.
  • the preparation of compounds of formula (I) of the present invention can be accomplished by the following illustrative methods and examples, which should not be construed in any way as limiting the scope of the invention.
  • the compounds of the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of methods known in the art and methods of the present invention are used.
  • the products obtained in each step of the reaction are obtained by separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, and the like.
  • the starting materials and chemical reagents required for the synthesis can be routinely synthesized or purchased according to the literature (available from SciFinder).
  • the heterocyclic compounds described in the general formula (I) of the present invention can be synthesized according to the following route: 1) amine compound A1 and intermediate B with a leaving group X (such as halogen) in a base (such as halogen) Potassium carbonate) catalyzed heating substitution reaction to obtain the target compound A4, of which the amine compound A1 can be purchased or synthesized according to literature methods (eg CN107739389).
  • the target compound A4 can also be obtained by performing a substitution reaction between compound A2 and intermediate B to obtain A3, and A3 is then heated to close the ring under the catalysis of an acid (such as TsOH), wherein compound A2 can be purchased or synthesized according to literature methods (such as WO2014025978).
  • Intermediate B can be synthesized according to the route shown below: 1) Starting material B1 and a starting material or reagent with a double leaving group X (such as halogen) under basic conditions (such as an organic base DIPEA or an inorganic base in DMF) Potassium carbonate) heating cyclization to generate intermediate B2; 2) the methanol solution of B2 is catalyzed by a palladium catalyst (such as PdCl 2 (dppf)) to carry out carbon monoxide intercalation reaction in carbon monoxide atmosphere to obtain intermediate ester B3, B3 can also be obtained by B4 and containing The raw materials or reagents of NH heterocycle A are obtained by substitution reaction under the catalysis of base (such as potassium carbonate); 3) The ester group in B3 is reduced by LAH to form intermediate alcohol B5; 4) B5 is further halogenated (such as using CBr 4 ) /PPh 3 or SOCl 2 ), thereby obtaining an important intermediate B.
  • the starting materials of the present invention can be synthesized according to methods known in the art, or can be purchased from chemical companies such as ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Shaoyuan Chemical Technology (Accela ChemBio Inc.), Beijing Coupling and other chemical companies .
  • the structures of the compounds of the present invention were determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS).
  • NMR nuclear magnetic resonance
  • MS mass spectrometry
  • reaction temperature is room temperature (20°C-30°C).
  • the reactions were carried out in an argon atmosphere or a nitrogen atmosphere.
  • Argon or nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon with a volume of about 1 L.
  • the hydrogen atmosphere means that after the reaction bottle is evacuated and then filled with hydrogen (repeated 3 times), a hydrogen balloon with a volume of about 1 L is connected.
  • the microwave reaction used a CEM Discover-SP type microwave reactor.
  • the compounds were purified by column chromatography or thin-layer chromatography, wherein the column chromatography used 200-300 mesh silica gel of Qingdao Ocean, and the thin-layer chromatography used GF254 silica gel plate of Qingdao Ocean with a thickness of 0.4-0.5 mm.
  • Column chromatography or thin layer chromatography developing solvent systems usually have a) dichloromethane and methanol system, b) petroleum ether and ethyl acetate system, or as shown in the examples.
  • the volume ratio of the solvent is adjusted according to the polarity of the compound, and can also be further adjusted by adding a small amount of triethylamine, or other acidic or basic reagents.
  • Compounds were also purified using a Waters mass spectrometry-guided automatic preparation system (mass detector: SQD2), with an appropriate acetonitrile/water (containing 0.1% trifluoroacetic acid or formic acid, or 0.05% ammonia water) gradient in 20 mL/water according to the polarity of the compound.
  • a reversed-phase high pressure column (XBridge-C18, 19 ⁇ 150 mm, 5 ⁇ m) was eluted at a flow rate of min.
  • Some of the examples can be purified using an automated prep system by adding 1N dilute hydrochloric acid, and then removing the solvent under reduced pressure to give the hydrochloride salt.
  • PdCl 2 refers to [1,1'-bis (diphenylphosphino) ferrocene] palladium dichloride.
  • LAH lithium aluminum hydride
  • DIPEA N,N-diisopropylethylamine
  • TsOH ⁇ H 2 O refers to p-toluenesulfonic acid monohydrate.
  • THF tetrahydrofuran
  • DMF refers to N,N-dimethylformamide
  • the characterization data for 18 are as follows:
  • the characterization data for 19 are as follows:
  • Compound 22 was synthesized according to the experimental procedures of steps 1 to 5 in compound 21, but in the third step 6-chloro-5-fluoronicotinonitrile was used instead of 2-cyano-5-fluoropyridine.
  • the experimental method is outlined as follows:
  • NCI-H929 (Nanjing Kebai, Cat. No. CBP60243) cells were cultured in RPMI 1640 complete medium [containing 10% FBS (GBICO, Cat. No. 10099-141) and 100 units/mL penicillin-streptomycin mixture (Thermo Fisher, Item No. 15140122)]. Cells (15,000 cells/mL) were seeded in 90 ⁇ L of complete medium in a 96-well plate. After overnight culture, 10 ⁇ L of compound solution was added to each well, and the cells were incubated at 37°C in a 5% CO 2 incubator for 6 days.
  • - Glo (CTG) kit Promega, Cat. No.
  • hPBMC peripheral blood mononuclear cells
  • the experimental method is outlined as follows:
  • Peripheral blood was collected from healthy volunteers with EDTA anticoagulant tubes, diluted with phosphate buffered saline (PBS) containing 2% fetal bovine serum (Gibco, Cat. No. 10099-141) in an equal volume, and then transferred 30 mL to the pre-added 15 mL density Gradient centrifuge (Sigma, Catalog No. 10771) was placed in a Sepmate-50 centrifuge tube (Stemcell, Catalog No. 86450), centrifuged at 1200 ⁇ g for 10 minutes at room temperature, and the PBMC-containing liquid in the upper layer was transferred to a new 50 mL centrifuge tube.
  • PBS phosphate buffered saline
  • Gibco Gibco, Cat. No. 10099-141
  • the compounds were dissolved and diluted in DMSO to 5mM (IC 50 values lower if the compound can reduce this initial concentration), followed by 4-fold dilution series to a minimum concentration of 0.31 ⁇ M with DMSO, and then each concentration point RPMI 1640 medium was diluted 50 times, and 10 ⁇ L was added to the cells in the above 96-well plate.
  • the 96 well plate was placed 37 °C, 5% CO 2 incubator After incubation of 1 hour, 10 ⁇ L a concentration of 100ng / mL of LPS (Sigma, Product Number L-2880), at 37 °C, 5% CO 2 in Continue to incubate overnight in the incubator. The supernatant was collected for the detection of TNF ⁇ .
  • TNF ⁇ and IL-2 were carried out according to the instructions of the respective ELISA kits (R&D, product numbers DY210 and DY202, respectively), and the OD450 value of each well was obtained. Taking the medium group containing 0.2% DMSO as 0% inhibition or stimulation, use XLfit software to draw compound inhibition or stimulation curves and calculate the corresponding IC 50 or EC 50 . The experimental results are shown in Table 1.
  • the experimental method is outlined as follows:
  • Compound solution The compound to be tested was dissolved in DMSO and made into a 10 mM stock solution, which was then diluted to 3 mM with DMSO, and then diluted to a 3 ⁇ M solution with extracellular fluid for subsequent use.
  • HEK293 cell line stably overexpressing the hERG potassium channel (Creacell, Cat. No. A-0320) was cultured in cells containing 10% fetal bovine serum (Gibco, Cat. No. 1428478) and 0.8 mg/mL G418 (Amresco, Cat. No. E859-5G). ) in DMEM medium (Gibco, product number 11995-065) at a temperature of 37°C and a carbon dioxide concentration of 5%. The old medium was removed and washed once with PBS (Gibco, Cat. No. 1009-141), then 1 mL of TrypLETM Express solution (Gibco, Cat. No.
  • Voltage stimulation protocol for whole-cell patch-clamp recording of whole-cell hERG potassium currents The cell membrane voltage was clamped at -80mV after the formation of the whole-cell seal. The clamping voltage was depolarized from -80mV to -50mV and held for 0.5 seconds (as leakage current detection), then stepped to 30mV and held for 2.5 seconds, and then quickly returned to -50mV and held for 4 seconds to excite the tail current of the hERG channel ( Peak tail current), hERG potassium current was recorded every 10 seconds. Experimental data were acquired with an EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA v2x73) software.
  • HEKA EPC-10 amplifier
  • PatchMaster HEKA v2x73
  • a capillary glass tube (Sutter Instruments) was drawn into a recording electrode using a microelectrode puller (Sutter Instruments).
  • Cell-laden coverslips were removed from the 24-well plate placed in the incubator and placed under an inverted microscope.
  • the recording electrode was perfused with the liquid in the electrode, and then the microelectrode manipulator (Sutter Instruments) was manipulated to touch the recording electrode to the cell surface and give negative pressure suction to form a G ⁇ seal; then perform rapid capacitance compensation; then continue to give negative pressure suction Aspirate until the cell membrane is broken to form a whole-cell recording mode.
  • test compound was dissolved in 5% DMA + 20% Solutol + 75% Saline vehicle to prepare a 0.5 mg/mL dosing solution.
  • Sprague-Dawley rats Three fed male Sprague-Dawley rats were given 2 mL/kg of the dosing solution by intravenous injection (IV) at a dose of 1 mg/kg, respectively, at 0.083, 0.25, 0.5, 1, 2, 4, 8 and Blood samples were collected 24 hours.
  • IV intravenous injection
  • Three other fed male Sprague-Dawley rats were given 10 mL/kg of the dosing solution by gavage (PO) at a dose of 5 mg/kg, respectively, at 0.25, 0.5, 1, 2, 4, 8, and 24 post-dose. Blood samples were collected every hour.

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Description

杂环类免疫调节剂 技术领域
本发明涉及作为免疫调节剂的杂环类化合物或其可药用的盐。本发明还涉及所述化合物或其可药用的盐的制备方法。本发明进一步涉及所述化合物或其可药用的盐在免疫调节、抗癌以及抗炎等方面的用途和使用方法。
背景技术
来那度胺等度胺类化合物是一种免疫调节剂(immunomodulatory drug；IMiD)，具有多种作用机制。在分子水平上，这类免疫调节剂通过结合E3泛素连接酶，调控CRL4 CRBN-E3泛素连接酶复合物的功能，引致Ikaros(IKZF1)、Aiolos(IKZF3)、蛋白激酶CK1α(CK1α)、转化终止因子GSPT1等泛素化而被26S蛋白酶体降解，从而改变多种细胞因子(比如IL-2、IL-6、IL-10、TNFα、IL-1β等)的分泌，影响免疫细胞活性。不同的度胺类化合物和CRL4 CRBN-E3泛素连接酶复合物结合后，表现出不同的底物蛋白降解特异性，因此具有不同的适应症。来那度胺和泊马度胺都能诱导cereblon(CRBN)降解在血液发展与分化过程中起重要作用的转录因子Ikaros和Aiolos，被用于治疗多发性骨髓瘤；但其中只有来那度胺能引起CK1α降解而被用于治疗5q缺失相关骨髓增生异常综合征。此外，CC-885能促进GSPT1的降解，而来那度胺和泊马度胺都无此作用。在非免疫调节方面，度胺类化合物可通过抑制血管内皮生长因子(VEGF)，而抑制肿瘤细胞的血管生成，也可直接抑制肿瘤细胞的增生及诱发异常细胞的分解。
通过免疫及非免疫的调节，度胺类化合物在多类恶性肿瘤和免疫性疾病的治疗中受到广泛关注，比如多发性骨髓瘤、骨髓增生异常综合症、血液瘤/实体瘤(比如淋巴瘤、非小细胞肺癌、胰脏、前列腺、脑部、肾脏、卵巢等)，以及系统性红斑狼疮等。此外，度胺类化合物还可与其它药物进行联合治疗疾病，比如与小分子靶向药和化疗药、大分子药物(比如PD-1抗体、CD20抗体、CD19抗体等)的联用。随着这类化合物在免疫调节、抗癌以及抗炎等方面研发的进一步开展，其临床适应症也在不断增加。
虽然度胺类化合物的化学结构相似，但各却具有不同的作用机理、临床治疗效果和毒副作用。沙利度胺在临床上的常见副作用，如神经疾病、便秘、嗜睡，在来那度胺却很少见；而泊马度胺则没有来那度胺的皮疹副作用。因此，需要开发新的免疫调节剂，扩展适应症，以满足不同临床需求。本发明涉及具有通式(I)所示结构的化合物，作为免疫 调节剂，通过与E3泛素连接酶结合而有效地调控IL-2、IL-6、IL-10、TNFα、VEGF等蛋白的表达和/或生物功能。
发明内容
定义
除非另有相反陈述，否则下列用在说明书和权利要求书中的术语具有下述含义。
“C x-y”表示碳原子数的范围，其中x和y均为整数，例如C 3-8环烷基表示具有3-8个碳原子的环烷基，即具有3、4、5、6、7或8个碳原子的环烷基。还应理解，“C 3-8”还包含其中的任意亚范围，例如C 3-7、C 3- 6、C 4-7、C 4-6、C 5-6等。
“烷基”指含有1至20个碳原子，例如1至8个碳原子、1至6个碳原子或1至4个碳原子的饱和的直链或支链的烃基基团。烷基的非限制性实例包括甲基、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-乙基丁基等。
“亚烷基”指含有1至20个碳原子，例如1至6个碳原子或1至4个碳原子的直链或支链的饱和烃的二价基团。亚烷基的非限制性实例包括-CH 2-、-CH(CH 3)-、-CH 2CH 2-、-CH 2CH 2CH 2-、-(CH 3)C(CH 3)-、-CH 2CH 2CH 2CH 2-、-CH 2CH(CH 3)CH 2-等。
“环烷基”指含有3至14个碳环原子的饱和环状烃基取代基。环烷基可以是单碳环，通常含有3至8个、3至7个或3至6个碳环原子。单环环烷基的非限制性实例包括环丙基、环丁基、环戊基、环己基、环庚基等。环烷基还可以是稠合、桥接或螺合到一起的双或三环，如十氢萘基、二环[2.2.2]辛烷、螺[3.3]庚烷等。
“杂环基或杂环”指饱和或部分不饱和的单环或多环环状基团，其包括3至20个环原子，例如可以是3至14个、3至12个、3至10个、3至8个、3至6个或5至6个环原子，其中一个或多个环原子选自氮、氧或S(O) m(其中m是整数0至2)，但不包括-O-O-、-O-S-或-S-S-的环部分，其余环原子为碳。优选包括3至12个环原子，更优选3至10个环原子，更优选4至7个环原子，更优选4至6个环原子，最优选5或6个环原子，其中1～4个是杂原子，更优选1～3个是杂原子，最优选1～2个是杂原子。单环杂环基的非限制性实例包含吡咯烷基、噁丁环基、哌啶基、哌嗪基、四氢呋喃基、四氢吡喃基、四氢噻喃基、吗啉基、硫代吗啉基、高哌嗪基、吖丁啶基等。多环杂环基包括稠、桥或螺多环杂环基，如八氢环戊二烯并[c]吡咯、八氢吡咯并[1,2-a]吡嗪、3,8-二氮杂二环[3.2.1]辛烷、5-氮杂螺[2.4]庚烷、2-氧杂-7-氮杂螺[3.5]壬烷等。
“芳基或芳环”指含有6至14个碳原子的芳香族单环或稠合多环基团，优选为6至10元，例如苯基和萘基，最优选苯基。所述芳基环可以稠合于杂芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为芳基环，非限制性实例包括：
Figure PCTCN2021102873-appb-000001
Figure PCTCN2021102873-appb-000002
等。
“杂芳基或杂芳环”指包含5至14个环原子的杂芳族体系，其中1至4个环原子选自包括氧、硫和氮的杂原子。杂芳基优选为5至10元，更优选杂芳基是5元或6元，例如呋喃基、噻吩基、吡啶基、吡咯基、嘧啶基、吡嗪基、吡唑基、咪唑基、四唑基、噁唑基、异噁唑基、噻唑基、异噻唑基、喹啉基、异喹啉基、吲哚基、异吲哚基等。所述杂芳基环可以稠合于芳基、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，非限制性实例包括：
Figure PCTCN2021102873-appb-000003
Figure PCTCN2021102873-appb-000004
等。
“卤素”指氟、氯、溴或碘。
“氰基”指-CN。
“氧代”指＝O。
“羰基”指-C(O)-基团。
“磺酰基”指-S(O) 2-基团。
“亚硫酰基”指-S(O)-基团。
“任选”意味着随后所描述的事件或环境可以但不必发生，该表述包括该事件或环境发生或不发生的情形。例如，“任选被烷基取代的杂环基团”意味着烷基可以但不必须存在，该表述包括杂环基团被烷基取代的情形和杂环基团不被烷基取代的情形。
“取代”指基团中的一个或多个氢原子，优选为5个，更优选为1～3个氢原子彼此独立地被相应数目的取代基取代。不言而喻，取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。所述取代基包括但不限于卤素、氰基、硝基、氧代、-SF 5、C 1-4烷基、C 3-7环烷基、4-7元杂环基、苯基、5-6元杂芳基等。
“异构体”指具有相同分子式但其原子结合的性质或顺序或其原子的空间排列不同的化合物。原子空间排列不同的异构体称为“立体异构 体”。立体异构体包括旋光异构体、几何异构体和构象异构体。
本发明的化合物可以以旋光异构体形式存在。旋光异构体包括对映异构体和非对映异构体。对映异构体是指两个立体异构体彼此镜像但相互不可重叠。外消旋混合物或外消旋体是指手性分子的左右手对映异构体数量相等的混合物。非对映异构体是指两个立体异构体不是彼此的镜像且不可重叠。当旋光异构体为单一异构体且其绝对构型确定，根据手性碳原子上取代基的构型，其为“R”或“S”的绝对构型；当旋光异构体的绝对构型未确定，依据所测的旋光值，其为(+)或(-)。制备和分离光学异构体的方法是本领域中已知的。
本发明的化合物也可以存在几何异构体。本发明考虑由碳-碳双键、碳-氮双键、环烷基或杂环基团周围的取代基的分布所产生的各种几何异构体和其混合物。碳-碳双键或碳-氮键周围的取代基指定为Z或E构型，环烷基或杂环周围的取代基指定为顺式或反式构型。
本发明的化合物还可能显示互变异构现象，例如酮-烯醇互变异构。
应该理解，本发明包括任何互变异构或立体异构形式和其混合物，并且不仅仅限于化合物的命名或化学结构式中所使用的任何一个互变异构或立体异构形式。
“同位素”是指在本发明化合物中出现的原子的所有同位素。同位素包括具有相同原子序数但不同质量数的那些原子。适合并入本发明化合物中的同位素的实例是氢、碳、氮、氧、磷、硫、氟和氯，分别例如但不限于 2H(D)、 3H、 13C、 14C、 15N、 17O、 18O、 31P、 32P、 35S、 18F和 36Cl。本发明的同位素标记化合物通常可通过本领域技术人员已知的传统技术或通过与所附实施例中描述的那些类似的方法使用适当的同位素标记的试剂代替非同位素标记的试剂来制备。这样的化合物具有各种潜在用途，例如作为测定生物活性中的标样和试剂。在稳定同位素的情况下，这样的化合物具有有利地改变生物、药理学或药代动力学性质的潜力。氘(D)为本发明的优选同位素，比如甲基、亚甲基或次甲基中的氢可被氘取代。
本发明的化合物可以以前药的形式给予。“前药”是指在活体内的生理条件下例如通过氧化、还原、水解等(它们各自利用酶或在没有酶参与下进行)转化成本发明的生物活性化合物的衍生物。前药的实例有下述化合物：其中本发明的化合物中的氨基被酰化、烷基化或磷酸化，例如二十烷酰基氨基、丙氨酰氨基、新戊酰氧基甲基氨基，或其中羟基被酰化、烷基化、磷酸化或转化成硼酸盐，例如乙酰氧基、棕榈酰氧基、新戊酰氧基、琥珀酰氧基、富马酰氧基、丙氨酰氧基，或其中羧基被酯化或酰胺化，或其中巯基与选择性地向靶和/或向细胞的胞质溶胶递送药物的载体分子，例如肽形成二硫桥键。这些化合物可以由本发明的化合物根据公知方法制备。
“可药用的盐”或者“药学上可接受的盐”是指由可药用的碱或酸，包 括无机碱或酸和有机碱或酸制成的盐。在本发明的化合物含有一个或多个酸性或碱性基团的情况下，本发明还包含它们相应的可药用盐。因此，含有酸性基团的本发明的化合物可以以盐形式存在并可根据本发明使用，例如作为碱金属盐、碱土金属盐或作为铵盐。这样的盐的更确切实例包括钠盐、钾盐、钙盐、镁盐或与氨或有机胺，例如乙胺、乙醇胺、三乙醇胺或氨基酸的盐。含有碱性基团的本发明的化合物可以以盐形式存在并可根据本发明以它们与无机或有机酸的加成盐的形式使用。合适的酸的实例包括盐酸、氢溴酸、磷酸、硫酸、硝酸、甲磺酸、对甲苯磺酸、萘二磺酸、草酸、乙酸、酒石酸、乳酸、水杨酸、苯甲酸、甲酸、丙酸、特戊酸、丙二酸、琥珀酸、庚二酸、富马酸、马来酸、苹果酸、氨基磺酸、苯基丙酸、葡糖酸、抗坏血酸、异烟酸、柠檬酸、己二酸和本领域技术人员已知的其它酸。如果本发明的化合物在分子中同时含有酸性和碱性基团，本发明除所提到的盐形式外还包括内盐或内铵盐。各盐可通过本领域技术人员已知的常规方法获得，例如通过在溶剂或分散剂中使这些与有机或无机酸或碱接触或通过与其它盐阴离子交换或阳离子交换。
“药物组合物”指含有一种或多种本发明所述的化合物或其可药用的盐、稳定同位素衍生物、异构体、前药及其混合物形式以及其他组分例如可药用的载体和赋形剂的组合物。药物组合物的目的是促进对生物体的给药，利于活性成分的吸收进而发挥生物活性。
因此，在本申请中当提及“化合物”、“本发明化合物”或“本发明所述化合物”时，包括所有所述化合物形式，例如其可药用的盐、稳定同位素衍生物、异构体、前药及其混合物。
“癌症/肿瘤”包括但不限于消化道/胃肠道癌、结肠癌、肝癌、皮肤癌(包括肥大细胞瘤和鳞状细胞癌)、乳腺癌、卵巢癌、前列腺癌、淋巴瘤、白血病(包括急性骨髓性白血病和慢性髓细胞性白血病)、肾癌、肺癌、肌肉癌、骨癌、膀胱癌、脑癌、黑素瘤(包括口腔和转移性黑素瘤)、卡波西肉瘤(包括多发性骨髓瘤的骨髓瘤)、骨髓增殖性疾病、增殖性糖尿病视网膜病变、血管增生相关病症/肿瘤等。
“炎性疾病”包括但不限于关节炎、桥本氏甲状腺炎、自身免疫性溶血性贫血、恶性贫血的自身免疫性萎缩性胃炎、自身免疫性脑脊髓炎、自身免疫性睾丸炎、古德帕斯丘病、自身免疫性血小板减少症、交感性眼炎、重症肌无力、格雷夫斯病、原发性胆汁性肝硬化、肝炎、原发性硬化性胆管炎、慢性侵袭性肝炎、非酒精性脂肪性肝病、非酒精性脂肪性肝炎、溃疡性结肠炎、膜性肾小球病、系统性红斑狼疮、类风湿性关节炎、牛皮癣性关节炎、干燥综合征、赖特综合征、多肌炎、皮肌炎、I-型干扰素病包括Aicardi-Goutières综合征和其他过度表达I-型干扰素的系统性硬化症、孟德尔疾病、结节性多动脉炎、多发性硬化症、复发缓解型多发性硬化症、原发性进行性多发性硬化症、继发性进展性多发 性硬化症、大疱性类天疱疮；另外基于O-细胞(体液)或T-细胞的自身免疫性疾病，包括科根综合征、强直性脊柱炎、韦格纳肉芽肿病、自身免疫性脱发、I-型或青少年型糖尿病、甲状腺炎。
“治疗有效量”是指包括可有效治疗或防止所述疾病的本发明化合物的量。
“患者”是指哺乳动物，尤其是人类。
本发明涉及一种通式(I)所示的化合物，或其可药用的盐、稳定同位素衍生物、异构体及其前药：
Figure PCTCN2021102873-appb-000005
其中：
环A为含N的4-10元杂环；
R 1为D或卤素；
R 2和R 3各自独立地选自H、D、卤素、氰基、C 1-6烷基、C 3-6环烷基或3-8元杂环基，但两者不可同时为H和/或D，或者R 2和R 3与其相连的碳原子共同组成C 3-6环烷烃或3-8元杂环；
R 4为D、卤素、氰基、氧代、C 1-6烷基、C 3-6环烷基、-OC 1-6烷基、-C(O)C 1-6烷基、-C(O)C 3-6环烷基、-S(O) 2C 1-6烷基、-S(O) 2C 3-6环烷基、芳基或杂芳基，其中所述烷基、环烷基、芳基和杂芳基的一个或多个氢任选被D、卤素、氰基、C 1-2烷基或氟代C 1-2烷基所取代；
m为整数0-4；且
n为整数0-2。
在一个实施方案中，环A为含一个N原子的4-10元单环杂环，或稠合、桥接或螺双环杂环(比如吗啉、哌啶、硫代吗啉-1,1-二氧化物、2-氧杂-5-氮双环[2.2.1]庚烷等)；R 4为H、D、卤素、氧代、-CF 3或C 1- 6烷基。
在一个实施方案中，环A为含两个N原子的6-10元单杂环，或稠合、桥接或螺双环杂环(比如哌嗪、3,6-二氮杂二环[3.1.1]庚烷、2,6-二氮杂螺[3.3]庚烷等)；R 4为与第二个N原子相连的C 1-6烷基、C 3-6环烷基、-C(O)C 1-6烷基、-C(O)C 3-6环烷基、-S(O) 2C 1-6烷基、-S(O) 2C 3-6环烷基、苯基或5-6元含N、O和/或S的杂芳基，其中所述烷基、环烷基、苯基和杂芳基的一个或多个氢任选被D、卤素、氰基、C 1-2烷基或氟代C 1-2烷基所取代。
在一个优选实施方案中，环A为哌嗪；R 4为与哌嗪的第二个N原 子相连的C 1-6烷基、C 3-6环烷基、-C(O)C 1-6烷基、-C(O)C 3-6环烷基、-S(O) 2C 1-6烷基、苯基、吡啶基或嘧啶基，其中所述烷基、环烷基、苯基、吡啶基或嘧啶基的一个或多个氢任选被D、卤素、氰基或C 1-2烷基所取代；n为0或1。
在一个实施方案中，R 2为H；R 3为氰基、C 1-6烷基或C 3-6环烷基。
在一个实施方案中，R 2和R 3都为甲基。
在一个实施方案中，R 2和R 3与其相连的碳原子共同组成C 3-6环烷烃。
在一个优选实施方案中，m为0。
在一些实施方案中，所述通式(I)化合物具有以下通式(II)所示的结构：
Figure PCTCN2021102873-appb-000006
其中：
R 5为H、C 1-6烷基、C 3-6环烷基、-C(O)C 1-6烷基、-C(O)C 3-6环烷基、-S(O) 2C 1-6烷基、苯基或5-6元含N、O和/或S的杂芳基，其中所述烷基、环烷基、苯基和杂芳基的一个或多个氢任选被D、卤素、氰基或C 1- 2烷基所取代。
在一个优选实施方案中，R 5为苯基、吡啶基或嘧啶基，其中所述苯基、吡啶基和嘧啶基的一个或两个氢任选被F或氰基所取代。
本发明进一步涉及以下化合物1-22，或其可药用的盐、稳定同位素衍生物、异构体、前药及其混合物：
Figure PCTCN2021102873-appb-000007
Figure PCTCN2021102873-appb-000008
Figure PCTCN2021102873-appb-000009
Figure PCTCN2021102873-appb-000010
Figure PCTCN2021102873-appb-000011
本发明的化合物能够有效抑制NCI-H929细胞的增殖，优选其IC 50小于50nM，更优选IC 50小于10nM。本发明的化合物对人PBMC细胞中TNFα的分泌具有显著抑制效应，优选其IC 50小于20nM，更优选IC 50小于10nM。本发明的化合物也对人PBMC细胞中IL-2的分泌具有显著刺激作用，优选其EC 50小于50nM，更优选EC 50小于10nM。
本发明还涉及一种药物组合物，所述药物组合物包含通式(I)所示的化合物或其可药用的盐、稳定同位素衍生物、异构体、前药及其混合物和一种或多种药学上可接受的载体或辅料。
本发明的一方面提供通式(I)所示的化合物或其可药用的盐、稳定同位素衍生物、异构体、前药及其混合物，或所述药物组合物用于调控或抑制E3泛素连接酶的活性，从而影响包括但不限于Aiolos、Ikaros、 Helios、CK1α、GSPT1、IL-2、IL-6、TNFα、IFNγ、VEGF等蛋白的表达和/或生物功能。
本发明的另一方面提供一种治疗或者预防Aiolos、Ikaros、Helios、CK1α、GSPT1、IL-2、IL-6、TNFα、IFNγ、VEGF等介导的相关疾病的方法，所述方法包括给予有需要的患者治疗有效量的通式(I)所示的化合物或其可药用的盐、稳定同位素衍生物、异构体、前药及其混合物，或包含所述化合物的药物组合物，所述疾病包括但不限于血液瘤(比如多发性骨髓瘤、淋巴瘤、白血病等)、实体瘤(比如肺癌、前列腺癌、头颈癌、乳腺癌、卵巢癌、子宫癌、胰脏癌、结肠癌、直肠癌、胃癌、食道癌、脑癌、肝癌、肾癌、皮肤癌、上皮癌、膀胱癌、成神经细胞瘤等)、自身免疫性疾病(比如系统性红斑狼疮、银屑病、炎性肠病等)、炎症(比如类风湿性关节炎)、神经退行性疾病(比如多发性硬化、阿尔茨海默氏病、帕金森氏病)、纤维化(比如肺纤维化)、皮肤疾病(比如黑色素瘤)、眼病、慢性阻塞性肺部疾病等，尤其是多发性骨髓瘤、淋巴瘤、骨髓增生异常综合征、系统性红斑狼疮、实体瘤等。
本发明还涉及一种药物组合物，所述药物组合物包含通式(I)所示的化合物或其可药用的盐、稳定同位素衍生物、异构体、前药及其混合物和至少一种额外的药物，其中所述至少一种额外的药物可为小分子化学治疗剂(比如NSAIDs、类固醇抗炎药、激酶靶向药、细胞毒类药物、DNA损伤相关药物等)或大分子免疫和/或炎症调节剂(比如PD-1抗体、CD20抗体、CD19抗体、TNFα抗体、IL-6抗体等)。
根据本发明，所述药物可以是任何药物剂型，包括但不限于片剂、胶囊剂、溶液剂、冻干制剂、注射剂。
本发明的药物制剂可以以每剂量单位包含预定量的活性成分的剂量单位形式给药。这种单位可根据治疗的病症、给药方法和患者的年龄、体重和状况包含例如0.1毫克至500毫克，优选0.5毫克至100毫克的本发明的化合物。此外，可以使用制药领域中所知的方法制备这种类型的药物制剂，比如把活性成分与一种或多种辅料和/或佐剂混合。
本发明药物制剂可适于通过任何所需的合适方法给药，例如通过经口(包括口腔或舌下)、直肠、经鼻、局部(包括口腔、舌下或经皮)、阴道或肠道外(包括皮下、肌内、静脉内或皮内)方法给药。
本发明还提供制备所述化合物的方法。本发明通式(I)所述化合物的制备可通过以下示例性方法和实施例完成，但这些方法和实施例不应以任何方式被认为是对本发明范围的限制。也可通过本领域技术人员所知的合成技术合成本发明所述的化合物，或者综合使用本领域已知方法和本发明所述方法。每步反应所得的产物用本领域已知的分离技术得到，包括但不限于萃取、过滤、蒸馏、结晶、色谱分离等。合成所需的起始物料和化学试剂可以根据文献(可从SciFinder上查询)常规合成或购买。
合成方法
本发明通式(I)所述的杂环类化合物可按照以下所示的路线合成：1)度胺类化合物A1与带有离去基团X(比如卤素)的中间体B在碱(比如碳酸钾)催化下加热取代反应得到目标化合物A4，其中度胺类化合物A1可购买或按照文献方法合成(比如CN107739389)。本目标化合物A4还可由化合物A2先与中间体B进行取代反应得到A3，A3再在酸(比如TsOH)催化下加热关环而得到，其中化合物A2可购买或按照文献方法合成(比如WO2014025978)。
Figure PCTCN2021102873-appb-000012
中间体B可按照以下所示的路线合成：1)起始原料B1与带有双离去基团X(比如卤素)的原料或试剂在碱性条件下(比如DMF中有机碱DIPEA或无机碱碳酸钾)加热环合生成中间体B2；2)B2的甲醇溶液在钯催化剂(比如PdCl 2(dppf))催化下在一氧化碳气氛中进行插羰反应得到中间体酯B3，B3也可由B4与含NH杂环A的原料或试剂在碱(比如碳酸钾)催化下进行取代反应而得到；3)B3中的酯基被LAH还原生成中间体醇B5；4)B5进一步被卤化(比如使用CBr 4/PPh 3或SOCl 2)，从而得到重要中间体B。
Figure PCTCN2021102873-appb-000013
实施例
本发明的起始原料可以按照本领域已知的方法来合成，或可购自ABCR GmbH&Co.KG、Acros Organics、Aldrich Chemical Company、韶远化学科技(Accela ChemBio Inc.)、北京偶合等化学品公司。
本发明化合物的结构是通过核磁共振(NMR)和/或质谱(MS)来确定的。NMR的测定使用Bruker ASCEND-400核磁仪，所用溶剂为氘代二甲基亚砜(DMSO-d 6)、氘代氯仿(CDC1 3)或氘代甲醇(CD 3OD)等，内标为四甲基甲硅烷(TMS)，化学位移是以10 -6(ppm)作为单位给出。MS的测定使用Agilent SQD(ESI)质谱仪(安捷伦6120)。
HPLC使用安捷伦1260DAD高压液相色谱仪(Poroshell 120EC-C18，50×3.0mm，2.7μm色谱柱)或Waters Arc高压液相色谱仪(Sunfirc C18，150×4.6mm，5μm色谱柱)。
实施例中如无特殊说明，反应温度为室温(20℃-30℃)。
实施例中如无特殊说明，反应均在氩气气氛或氮气气氛下进行。氩气气氛或氮气气氛是指反应瓶连接一个约1L容积的氩气或氮气气球。
氢气气氛是指反应瓶在抽真空再充入氢气(反复3次)后，连接一个约1L容积的氢气气球。
微波反应使用CEM Discover-SP型微波反应器。
实施例中的反应进程的监测使用安捷伦的液质联用色谱仪(1260/6120)，也可采用薄层层析(TLC)，所用硅胶板的厚度为0.15～0.2mm(青岛海洋GF254)。
化合物的纯化采用柱层析或薄层层析，其中柱层析使用青岛海洋的200～300目硅胶，薄层层析使用青岛海洋的厚度为0.4～0.5mm的GF254硅胶板。
柱层析或薄层层析展开溶剂体系通常有a)二氯甲烷和甲醇体系，b)石油醚和乙酸乙酯体系，或如实施例中所示。溶剂的体积比根据化合物的极性不同而进行调节，也可以加入少量的三乙胺、或其它酸性或碱性试剂进一步调节。
化合物的纯化还采用Waters的质谱导向自动制备系统(质谱检测器：SQD2)，根据化合物的极性用适当的乙腈/水(含0.1％三氟乙酸或甲酸，或0.05％氨水)梯度于20mL/min的流速洗脱反相高压柱(XBridge-C18，19×150mm，5μm)。部分实施例可在使用自动制备系统纯化后加入1N稀盐酸，然后减压除去溶剂，得到盐酸盐。
缩写PdCl 2(dppf)是指[1,1'-双(二苯基膦基)二茂铁]二氯化钯。
缩写LAH是指氢化铝锂。
缩写DIPEA是指N,N-二异丙基乙胺。
缩写TsOH·H 2O是指对甲基苯磺酸一水合物。
缩写THF是指四氢呋喃。
缩写DMF是指N,N-二甲基甲酰胺。
实施例1
3-(4-((4-(1-吗啉代乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物1)
Figure PCTCN2021102873-appb-000014
第一步
3-(1-氧代-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)异吲哚啉-2-基)哌啶-2,6-二酮(1b)
室温下将3-(7-氨基-3-氧代-1H-异吲哚-2-基)哌啶-2,6-二酮1a(5g，19.3mmol)、联硼酸频那醇酯(7.35g，29.0mmol)和乙腈(100mL) 混合，然后加入亚硝酸叔丁酯(2.19g，21.3mmol)。将混合物在室温搅拌18小时，然后减压除去溶剂。残余物用硅胶柱层析(二氯甲烷/甲醇＝100/1)纯化，得到目标产物1b(3.9g,55％)。
MS m/z(ESI):371[M+1]
第二步
(2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)硼酸(1c)
室温下将1b(3.9g，10.5mmol)、THF(100mL)和水(20mL)混合，然后缓慢地加入偏高碘酸钠(6.7g，31.6mmol)。将混合物在室温下搅拌2小时，然后加入稀盐酸(1N，7.3mL，7.3mmol)。将混合物在室温下搅拌18小时后，减压除去溶剂，残余物溶于二氯甲烷(20mL)和水(20mL)。室温搅拌18小时后过滤，得到目标产物1c(2.3g，76％)。
MS m/z(ESI):289[M+1]
第三步
3-(4-羟基-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(1d)
氮气气氛下向(1c(1g，3.5mmol)的二甲基亚砜(20mL)溶液中加入过氧化氢水溶液(2.76mL，27mmol)。将混合物在室温搅拌18小时后，加入水(30mL)，然后用乙酸乙酯(3×50mL)萃取。合并的有机相用饱和食盐水(3×50mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/1至85/15)纯化，得到目标产物1d(110mg，12％)。
MS m/z(ESI):261[M+1]
第四步
4-(1-吗啉代乙基)苯甲酸甲酯(1f)
室温下将4-(1-溴乙基)苯甲酸甲酯1e(242mg，1mmol)、吗啉(87mg，1mmol)、碳酸钾(151.8mg，1.1mmol)和乙腈(1mL)混合并搅拌18小时，然后加入水(3mL)并用乙酸乙酯(3×3mL)萃取。合并的有机相用水(3×2mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，得到目标产物1f(280mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):250[M+1]
第五步
(4-(1-吗啉代乙基)苯基)甲醇(1g)
将1f(280mg，粗品)溶于THF(2mL)并冷却到0℃，然后在氮气气氛下加入LAH(128mg，3.37mmol)。0℃下搅拌1小时后，然后在室温下再搅拌3小时。向混合物中加入水(2mL)，然后用乙酸乙酯(3×3mL)萃取。合并的有机相用水(3×2mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/1至1/1)纯化，得到目标产物1g(160mg， 73％)。
MS m/z(ESI):222[M+1]
第六步
4-(1-(4-(溴甲基)苯基)乙基)吗啉(1h)
将1g(80mg，0.36mmol)、四溴化碳(143mg，0.43mmol)和二氯甲烷(5mL)混合，然后在氮气气氛下加入三苯基膦(114mg，0.43mmol)。室温下搅拌48小时后，减压除去溶剂，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/1至7/3)纯化，得到目标产物1h(20mg，20％)。
MS m/z(ESI):284[M+1]
第七步
3-(4-((4-(1-吗啉代乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(1)
室温下将1h(20mg，0.07mmol)、1d(18.4mg，0.07mmol)、碘化钾(17.6mg，0.11mmol)、碳酸钾(14.6mg，0.11mmol)和乙腈(1mL)混合，加热到80℃并搅拌18小时。冷却到室温后，减压除去溶剂，残余物用反相制备高效液相色谱纯化，得到目标产物1(6.1mg，固体，19％)。
MS m/z(ESI):464[M+1]
1H NMR(400MHz,CD 3OD)δ7.41-7.36(m,3H),7.32-7.27(m,3H),7.19(d,J＝8.0Hz,1H),5.14(s,2H),5.06-5.02(m,1H),4.35(q,J＝17.4Hz,3H),3.63-3.57(m,4H),2.80-2.75(m,1H),2.72-2.67(m,1H),2.48-2.34(m,4H),2.12-2.01(m,2H),1.36(d,J＝6.7Hz,3H).
实施例2
3-(4-((4-(1-吗啉代环丙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物2)
Figure PCTCN2021102873-appb-000015
第一步
4-(1-吗啉代环丙基)苯甲酸甲酯(2b)
室温下将4-(1-氨基环丙基)苯甲酸甲酯2a(191mg，1mmol)、1-溴-2-(2-溴乙氧基)乙烷(696mg，3mmol)、DIPEA(645mg，5mmol)和DMF(3mL)混合，加热到110℃并搅拌18小时。冷却到室温后，加入水(10mL)，然后用乙酸乙酯(3×10mL)萃取。合并的有机相 用水(3×5mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/1至19/1)纯化，得到目标产物2b(150mg，57％)。
MS m/z(ESI):262[M+1]
第二步
(4-(1-吗啉代环丙基)苯基)甲醇(2c)
0℃下向2b(150mg，0.57mmol)的THF(5mL)溶液中加入LAH(43.6mg，1.15mmol)，室温下搅拌2小时后，加入水(2mL)，然后用乙酸乙酯(3×10mL)萃取。合并的有机相用水(3×5mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，得到目标产物2c(110mg，84％)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):234[M+1]
第三步
4-(1-(4-(氯甲基)苯基)环丙基)吗啉(2d)
室温下向2c(110mg，0.47mmol)的二氯甲烷(5mL)溶液中加入氯化亚砜(84mg，0.71mmol)。室温下搅拌2小时后，减压除去溶剂，得到目标产物2d(100mg，20％)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):252[M+1]
第四步
3-(4-((4-(1-吗啉代环丙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(2)
室温下将2d(100mg，0.4mmol)、1d(52mg，0.2mmol)、碘化钾(66mg，0.4mmol)、碳酸钾(55mg，0.4mmol)和乙腈(5mL)混合，加热到100℃并搅拌18小时。冷却到室温后过滤，滤液在减压条件下浓缩至干，残余物用反相制备高效液相色谱纯化，得到目标产物2(5.2mg，固体，5.5％)。
MS m/z(ESI):476[M+1]
1H NMR(400MHz,CD 3OD)δ7.48(t,J＝7.3Hz,3H),7.41(d,J＝7.3Hz,3H),7.29(d,J＝7.9Hz,1H),5.25(s,2H),5.14(dd,J＝13.3,5.1Hz,1H),4.45(q,J＝17.3Hz,2H),3.73-7.55(m,4H),2.91-2.85(m,1H),2.81-2.64(m,4H),2.50(dd,J＝13.2,4.7Hz,1H),2.20-2.13(m,1H),1.31-1.23(m,1H),1.15-1.03(m,2H),0.98-0.88(m,2H).
实施例3
3-(4-((4-((R)-1-吗啉代乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物3)
Figure PCTCN2021102873-appb-000016
第一步
(R)-4-(1-(4-溴苯基)乙基)吗啉(3b)
将(R)-1-(4-溴苯基)乙胺3a(2g，10mmol)溶解在DMF(40mL)中，然后加入碳酸钾(4.15g，30mmol)和1-溴-2-(2-溴乙氧基)乙烷(2.58g，11.1mmol)。将混合物在室温搅拌3小时，然后加热至80℃并搅拌3小时。冷却到室温后，补加1-溴-2-(2-溴乙氧基)乙烷(258mg，1.11mmol)，加热到80℃并搅拌2小时。冷却到室温后，用水(250mL)稀释，然后用乙酸乙酯(2×150mL)萃取。合并的有机相依次用饱和碳酸氢钠溶液(2×150mL)和饱和食盐水(150mL)洗涤，然后经无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至96/4)纯化，得到目标产物3b(2.14g，79％)。
MS m/z(ESI):270[M+1]
1H NMR(400MHz,CDCl 3)δ7.44(d,J＝8.3Hz,2H),7.21(d,J＝8.0Hz,2H),3.77-3.60(m,4H),3.27(d,J＝4.6Hz,1H),2.46(s,2H),2.36(s,2H),1.30(t,J＝9.0Hz,3H).
第二步
(R)-4-(1-吗啉代乙基)苯甲酸甲酯(3c)
将3b(2.14g，7.92mmol)溶于甲醇(80mL)，加入三乙胺(4g，39.6mmol)并冷却至-40℃。用一氧化碳气体鼓泡30分钟，然后加入PdCl 2(dppf)(1.16g，1.58mmol)。将混合物在密封管中加热至100℃并搅拌20小时。冷却到室温后过滤，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至3/7)纯化，得到目标产物3c(1.7g，86％)。
MS m/z(ESI):250[M+1]
1H NMR(400MHz,CDCl 3)δ7.99(d,J＝4.7Hz,2H),7.41(s,2H),3.91(s,3H),3.69(s,4H),3.36(s,1H),2.49(s,2H),2.35(s,2H),1.34(s,3H).
第三步
(R)-(4-(1-吗啉代乙基)苯基)甲醇(3d)
将3c(1.7g，6.82mmol)溶于无水THF(40mL)并冷却至0℃，然后加入LAH(600mg，15.8mmol)。将混合物在室温下搅拌16小时后，冷却至0℃，然后用NaOH溶液(1N，8mL)淬灭。过滤后，滤 液经无水硫酸钠干燥，然后再次过滤，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至98/2)纯化，得到目标产物3d(1.45g，96％)。
MS m/z(ESI):222[M+1]
1H NMR(400MHz,CDCl 3)δ7.33(s,4H),4.68(s,2H),3.70(s,4H),3.33(s,1H),2.50(s,2H),2.39(s,2H),1.72(s,1H),1.37(s,3H).
第四步
(R)-4-(1-(4-(氯甲基)苯基)乙基)吗啉(3e)
将3d(98mg，0.44mmol)溶于二氯甲烷(10mL)，然后加入氯化亚砜(1mL)。室温下搅拌2小时后，浓缩至干，得到目标产物3e(120mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):240[M+1]
第五步
3-(4-((4-((R)-1-吗啉代乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮甲酸盐(3)
将3e(120mg，粗品)、碳酸钾(182mg，1.32mmol)、1d(92mg，0.35mmol)、碘化钾(73mg，0.44mmol)和乙腈(10mL)混合，加热到80℃并搅拌3小时。冷却到室温后，减压除去溶剂，残余物用水(20mL)稀释，然后用乙酸乙酯(2×20mL)萃取。合并的有机相经无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用反相制备高效液相色谱纯化，得到目标产物3(34.5mg，固体，19％)。
MS m/z(ESI):464[M+1]
1H NMR(400MHz,CD 3OD)δ8.29(s,1H),7.48(dd,J＝7.9,6.3Hz,3H),7.40(dd,J＝7.7,3.0Hz,3H),7.28(d,J＝8.0Hz,1H),5.23(s,2H),5.14(dd,J＝13.3,5.2Hz,1H),4.45(q,J＝17.4Hz,2H),3.71(t,J＝4.7Hz,4H),3.62(dd,J＝13.5,6.7Hz,1H),2.95-2.84(m,1H),2.80-2.66(m,3H),2.58-2.44(m,3H),2.16(dtd,J＝12.8,5.2,2.3Hz,1H),1.46(d,J＝6.8Hz,3H).
化合物4参照化合物3中第一步至第五步的实验步骤操作，但在第一步中用(S)-1-(4-溴苯基)乙胺代替3a。
Figure PCTCN2021102873-appb-000017
化合物4的核磁数据如下：
Figure PCTCN2021102873-appb-000018
Figure PCTCN2021102873-appb-000019
实施例4
(S)-3-(4-((4-(2-吗啉代丙烷-2-基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物5)
Figure PCTCN2021102873-appb-000020
第一步
3-羟基-2-甲基苯甲酸甲酯(5b)
将3-羟基-2-甲基苯甲酸5a(5g，33mmol)溶于甲醇(50mL)中，然后加入浓硫酸(980mg，9.9mmol)。将混合物加热至70℃并搅拌20小时。冷却到室温后，浓缩至约10mL，然后将残余物缓慢加入冷水(100mL)中，然后用饱和碳酸氢钠溶液调节至pH＝4并搅拌20分钟。过滤收集沉淀物，得到目标产物5b(4.48g，82％)。
MS m/z(ESI):167[M+1]
1H NMR(400MHz,DMSO-d 6)δ9.67(s,1H),7.17(d,J＝7.7Hz,1H),7.08(t,J＝7.8Hz,1H),6.99(d,J＝7.9Hz,1H),3.79(s,3H),2.27(s,3H).
第二步
3-((叔丁基二甲基甲硅烷基)氧基)-2-甲基苯甲酸甲酯(5c)
将5b(4.48g，27mmol)溶于DMF(25mL)，冷却到0℃并依次 加入咪唑(4.59g，67mmol)和叔丁基二甲基氯硅烷(4.47g，30mmol)。将混合物升温至室温并搅拌16小时，然后用水(125mL)稀释并用乙酸乙酯(2×100mL)萃取。合并的有机相用水(3×100mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至93/7)纯化，得到目标产物5c(7.57g，100％)。
MS m/z(ESI):281[M+1]
1H NMR(400MHz,CDCl 3)δ7.21(dd,J＝7.8,1.0Hz,1H),6.87(t,J＝7.9Hz,1H),6.71(dd,J＝8.0,0.9Hz,1H),3.66(s,3H),2.20(s,3H),0.83-0.77(m,9H),0.02–-0.04(m,6H).
第三步
2-(溴甲基)-3-((叔丁基二甲基甲硅烷基)氧基)苯甲酸甲酯(5d)
将5c(561mg，2mmol)溶于四氯化碳(30mL)中，加入N-溴代琥珀酰亚胺(374mg，2.1mmol)和过氧化二苯甲酰(41mg，0.2mmol)。将混合物加热至80℃并搅拌3小时。冷却到室温后过滤，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至97/3)纯化，得到目标产物5d(630mg，88％)。
1H NMR(400MHz,CDCl 3)δ7.52(dd,J＝7.8,1.2Hz,1H),7.23(t,J＝8.0Hz,1H),7.00(dd,J＝8.2,1.1Hz,1H),5.02(s,2H),3.93(s,3H),1.10–1.03(m,9H),0.33–0.26(m,6H).
第四步
(S)-5-氨基-4-(4-((叔丁基二甲基硅烷基)氧基)-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(5e)
将5d(630mg，1.75mmol)和(S)-4,5-二氨基-5-氧代戊酸叔丁酯盐酸盐(460mg，1.93mmol)溶于乙腈(5mL)中，然后加入DIPEA(566mg，4.38mmol)。将混合物加热至50℃并搅拌18小时。冷却到室温后，减压除去溶剂，残余物用硅胶柱层析(石油醚/乙酸乙酯＝44/56至0/100)纯化，得到目标产物5e(545mg，69％)。
MS m/z(ESI):449[M+1]
1H NMR(400MHz,CDCl 3)δ7.47-7.43(m,1H),7.38-7.32(m,1H),6.98-6.94(m,1H),6.33(s,1H),5.36-5.31(m,1H),4.91-4.85(m,1H),4.43-4.30(m,2H),2.48-2.09(m,4H),1.42(s,9H),1.03-0.98(m,9H),0.28-0.22(m,6H).
第五步
(S)-5-氨基-4-(4-羟基-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(5f)
室温下将5e(545mg，1.21mmol)溶于THF(3mL)中，加入四丁基氟化铵的THF溶液(1M，1.5mL，1.5mmol)并搅拌30分钟。将混合物用水(20mL)稀释，然后用乙酸乙酯(3×20mL)萃取。合并的有机相用无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝1/1至0/100)纯化，得到目 标产物5f(270mg，67％)。
MS m/z(ESI):335[M+1]
第六步
4-(2-(4-溴苯基)丙-2-基)吗啉(5h)
将2-(4-溴苯基)丙-2-胺5g(214mg，1mmol)溶于DMF(4mL)，加入碳酸钾(415mg，3mmol)和1-溴-2-(2-溴乙氧基)乙烷(278mg，1.2mmol)。将混合物加热到80℃并搅拌20小时。冷却到室温后，用水(80mL)稀释，用乙酸乙酯(3×40mL)萃取。合并的有机相依次用水(2×40mL)和盐水(40mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至3/2)纯化，得到目标产物5h(207mg，73％)。
MS m/z(ESI):284[M+1]
1H NMR(400MHz,CDCl 3)δ7.42(s,4H),3.73-3.61(m,4H),2.50-2.41(m,4H),1.31(s,6H).
第七步
4-(2-吗啉代丙烷-2-基)苯甲酸甲酯(5i)
将5h(207mg，0.73mmol)溶于甲醇(20mL)，然后加入三乙胺(369mg，3.64mmol)。冷却至-40℃后用一氧化碳气体鼓泡20分钟，然后加入PdCl 2(dppf)(107mg，0.15mmol)。将混合物在封管中加热至100℃并搅拌20小时。冷却到室温后，混合物通过硅藻土过滤。滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至1/1)纯化，得到目标产物5i(189mg，98％)。
MS m/z(ESI):264[M+1]
1H NMR(400MHz,CDCl 3)δ7.97(d,J＝8.5Hz,2H),7.62(d,J＝8.5Hz,2H),3.91(s,3H),3.71-3.63(m,4H),2.50-2.41(m,4H),1.35(s,6H).
第八步
(4-(2-吗啉代丙烷-2-基)苯基)甲醇(5j)
将5i(189mg，0.72mmol)溶于THF(20mL)并冷却至0℃，然后加入LAH(35mg，1.44mmol)。将混合物在室温下搅拌2小时，然后用NaOH溶液(1N，0.2mL)淬灭并过滤。滤液用无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝1/1至3/7)纯化，得到目标产物5j(77mg，46％)。
MS m/z(ESI):236[M+1]
第九步
4-(2-(4-(氯甲基)苯基)丙-2-基)吗啉(5k)
将5j(77mg，0.33mmol)溶于二氯甲烷(2mL)，然后加入氯化亚砜(0.5mL)。将混合物在室温下搅拌16小时，然后浓缩至干，得到目标产物5k(固体，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):254[M+1]
第十步
(S)-5-氨基-4-(4-((4-(2-吗啉代丙烷-2-基)苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氧戊酸叔丁酯(5l)
将5k(粗品，约0.33mmol)溶于DMF(2mL)，然后加入碳酸钾(138mg，1mmol)和5f(110mg，0.33mmol)。将混合物加热到50℃并搅拌24小时。冷却到室温后，用水(20mL)稀释，然后用乙酸乙酯(3×20mL)萃取。合并的有机相用水(3×20mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物通过硅胶柱层析(二氯甲烷/甲醇＝100/0至19/1)纯化，得到目标产物5l(163mg，两步90％)。
MS m/z(ESI):552[M+1]
第十一步
(S)-3-(4-((4-(2-吗啉代丙烷-2-基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮甲酸盐
将5l(163mg，0.3mmol)溶于乙腈(2mL)中，加入TsOH·H 2O(56mg，0.3mmol)。将混合物加热至90℃并搅拌18小时。冷却到室温后，再次加入TsOH·H 2O(56mg，0.3mmol)，然后加热至90℃并搅拌2小时。冷却至室温后，加入饱和碳酸氢钠溶液(20mL)，然后用乙酸乙酯(3×20mL)萃取。合并的有机相经无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用反相制备高效液相色谱纯化，得到目标产物5(37mg，固体，26％)。
MS m/z(ESI):478[M+1]
1H NMR(400MHz,CD 3OD)δ8.26(s,1H),7.62(d,J＝8.4Hz,2H),7.52-7.45(m,3H),7.42(d,J＝7.0Hz,1H),7.31(d,J＝7.6Hz,1H),5.25(s,2H),5.16(dd,J＝13.3,5.2Hz,1H),4.47(q,J＝17.3Hz,2H),3.75–3.67(m,4H),2.96-2.87(m,1H),2.82-2.76(m,1H),2.69–2.61(m,4H),2.57-2.46(m,1H),2.21-2.15(m,1H),1.50(s,6H).
实施例5
4-(4-((R)-1-(4-(((2-((S)-2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氧基)甲基)苯基)乙基)哌嗪-1-基)-3-氟苄腈(化合物6)
Figure PCTCN2021102873-appb-000021
第一步
(R)-1-(1-(4-溴苯基)乙基)-4-甲苯磺酰基哌嗪(6a)
将(R)-1-(4-溴苯基)乙胺3a(2g，10mmol)、N,N-双(2-氯乙基)-4-甲基苯磺酰胺(3.11g，10.5mmol)和DIPEA(2.58g，20mmol)放入30mL封管中，加热到125℃并搅拌18小时。冷却至室温后，加入乙醇(80mL)并搅拌，在搅拌的过程中逐渐滴加水(120mL)。将所得混合物搅拌30分钟后，过滤收集沉淀物，得到目标产物6a(3.97g，94％)。
MS m/z(ESI):423[M+1]
1H NMR(400MHz,CDCl 3)δ7.62(d,J＝8.1Hz,2H),7.40(d,J＝8.3Hz,2H),7.32(d,J＝8.0Hz,2H),7.11(d,J＝8.3Hz,2H),3.32(q,J＝6.6Hz,1H),2.97(s,4H),2.61-2.49(m,2H),2.49-2.36(m,2H),2.44(s,3H),1.27(d,J＝6.7Hz,3H).
第二步
(R)-1-(1-(4-溴苯基)乙基)哌嗪(6b)
将6a(1g，2.36mmol)溶解在三氟乙酸(2.69g，23.6mmol)中，然后加入浓硫酸(1.62g，16.5mmol)。将混合物加热至75℃并搅拌16小时。冷却至室温后，缓慢加入饱和碳酸氢钠溶液(100mL)，然后用乙酸乙酯(3×100mL)萃取。合并的有机相无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，得到目标产物6b(860mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):269[M+1]
第三步
(R)-4-(4-(1-(4-(溴代苯基)乙基)哌嗪-1-基)-3-氟苄腈(6c)
将6b(860mg，粗品，约2.36mmol)和3,4-二氟苄腈(328mg，2.36mmol)溶于DMF(15mL)中，然后加入碳酸钾(978mg，7.08mmol)。将混合物加热至110℃并搅拌16小时。冷却至室温后，加入 水(150mL)并搅拌1小时。通过过滤收集沉淀物，得到目标产物6c(506mg，55％)。
MS m/z(ESI):388[M+1]
1H NMR(400MHz,CDCl 3)δ7.45(d,J＝6.5Hz,2H),7.35(d,J＝8.0Hz,1H),7.29-7.15(m,3H),6.88(t,J＝8.6Hz,1H),3.39(d,J＝5.4Hz,1H),3.19(s,4H),2.63(s,2H),2.54(s,2H),1.36(d,J＝6.0Hz,3H).
第四步
(R)-4-(1-(4-(4-氰基-2-氟苯基)哌嗪-1-基)乙基)苯甲酸甲酯(6d)
将6c(506mg，1.3mmol)和甲醇(40mL)混合，然后加入三乙胺(659mg，6.5mmol)。冷却至-40℃后，用一氧化碳鼓泡20分钟，然后加入PdCl 2(dppf)(190mg，0.26mmol)。将混合物加热至100℃并搅拌18小时。冷却到室温后，将反应混合物通过硅藻土过滤。滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至1/1)纯化，得到目标产物6d(360mg，75％)。
MS m/z(ESI):368[M+1]
1H NMR(400MHz,CDCl 3)δ8.01(d,J＝7.3Hz,2H),7.42(d,J＝7.1Hz,2H),7.35(d,J＝8.1Hz,1H),7.25(d,J＝9.8Hz,1H),6.89(t,J＝8.4Hz,1H),3.92(s,3H),3.48(d,J＝5.5Hz,1H),3.20(s,4H),2.66(s,2H),2.54(s,2H),1.39(d,J＝5.7Hz,3H).
第五步
(R)-3-氟-4-(4-(1-(4-(羟甲基)苯基)乙基)哌嗪-1-基)苄腈(6e)
将6d(360mg，0.98mmol)溶于THF(20mL)，冷却至0℃，然后加入LAH(45mg，1.18mmol)。将混合物在室温下搅拌2小时，冷却至0℃，然后用NaOH溶液(1N，1mL)淬灭。将反应液用无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/乙酸乙酯＝7/3至4/1)纯化，得到目标产物6e(100mg，30％)。
MS m/z(ESI):340[M+1]
第六步
(R)-4-(4-(1-(4-(氯甲基)苯基)乙基)哌嗪-1-基)-3-氟苄腈(6f)
将6e(100mg，0.29mmol)溶于二氯甲烷(5mL)，然后加入氯化亚砜(69mg，0.58mmol)。将混合物在室温下搅拌16小时，浓缩至干，得到目标产物6f(油状物，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):358[M+1]
第七步
(S)-5-氨基-4-(4-((4-((R)-1-(4-(4-氰基-2-氟苯基)哌嗪-1-基)乙基)苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(6g)
将6f(粗品，约0.29mmol)溶于DMF(5mL)，然后加入5f(97mg，0.29mmol)和碳酸钾(200mg，1.45mmol)。将混合物加热至50℃ 并搅拌20小时。冷却到室温后，混合物用水(80mL)稀释，过滤收集沉淀物，得到目标产物6g(197mg，两步104％)。
MS m/z(ESI):656[M+1]
第八步
4-(4-((R)-1-(4-(((2-((S)-2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氧基)甲基)苯基)乙基)哌嗪-1-基)-3-氟苄腈(6)
将6g(197mg，0.29mmol)溶于乙腈(10mL)，然后加入TsOH·H 2O(110mg，0.58mmol)。将混合物加热至90℃并搅拌3小时。冷却至室温后，用饱和碳酸氢钠溶液(30mL)稀释，然后用乙酸乙酯(3×30mL)萃取。合并的有机相用无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用反相制备高效液相色谱纯化，得到目标产物(21mg，固体，12％)。
MS m/z(ESI):582[M+1]
1H NMR(400MHz,CD 3OD)δ7.50–7.34(m,8H),7.28(d,J＝8.0Hz,1H),7.05(t,J＝8.6Hz,1H),5.21(s,2H),5.12(dd,J＝13.4,5.2Hz,1H),4.43(q,J＝17.4Hz,2H),3.47(q,J＝6.7Hz,1H),3.25–3.15(m,4H),2.94–2.82(m,1H),2.75(ddd,J＝17.6,4.6,2.4Hz,1H),2.71–2.63(m,2H),2.58–2.41(m,3H),2.15(tdd,J＝7.6,5.8,3.1Hz,1H),1.41(d,J＝6.7Hz,3H).
化合物7、8、9参照化合物6中第一步至第八步的实验步骤合成，但在第一步中用不同的化合物代替(R)-1-(4-溴苯基)乙胺3a。
Figure PCTCN2021102873-appb-000022
化合物7、8、9的核磁数据如下：
Figure PCTCN2021102873-appb-000023
实施例6
2-(4-(((2-((S)-2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氧基)甲基)苯基)-2-吗啉代乙腈(化合物10)
Figure PCTCN2021102873-appb-000024
第一步
(S)-5-氨基-4-(4-((4-甲酰基苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(10a)
将5f(167mg，0.5mmol)和4-(氯甲基)苯甲醛(77mg，0.5mmol)溶于DMF(3mL)中，然后加入碳酸钾(207mg，1.5mmol)，加热到 50℃并搅拌18小时。冷却到室温后过滤，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至9/1)纯化，得到目标产物10a(164mg，72％)。
MS m/z(ESI):453[M+1]
第二步
(S)-4-(((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氧基)甲基)苯甲醛(10b)
将10a(164mg，0.36mmol)溶于乙腈(20mL)，然后加入TsOH·H 2O(138mg，0.72mmol)，加热至90℃并搅拌18小时。冷却到室温后，减压除去溶剂，残余物中加入用饱和碳酸氢钠溶液(20mL)，然后用乙酸乙酯(3×20mL)萃取。合并的有机相用无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至47/3)纯化，得到目标产物10b(48mg，35％)。
MS m/z(ESI):379[M+1]
第三步
2-(4-(((2-((S)-2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氧基)甲基)苯基)-2-吗啉代乙腈(10)
将10b(48mg，0.13mmol)和二氯甲烷(10mL)混合，然后加入吗啉(23mg，0.26mmol)，三甲基氰硅烷(26mg，0.26mmol)和三氟甲磺酸镱(III)(8mg，0.013mmol)。将混合物在室温下搅拌61小时，然后减压除去溶剂。残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至91/9)和薄层硅胶色谱(二氯甲烷/甲醇＝8/1)纯化，得到目标产物10(44.6mg，固体，72％)。
MS m/z(ESI):475[M+1]
1H NMR(400MHz,DMSO-d 6)δ10.97(s,1H),7.58(d,J＝8.2Hz,2H),7.52-7.46(m,3H),7.33(dd,J＝7.7,3.1Hz,2H),5.40(s,1H),5.28(s,2H),5.12(dd,J＝13.3,5.1Hz,1H),4.43(d,J＝17.5Hz,1H),4.27(d,J＝17.5Hz,1H),3.66-3.52(m,4H),2.96-2.85(m,1H),2.58(d,J＝19.0Hz,2H),2.48-2.34(m,4H),1.99(dd,J＝9.0,3.7Hz,1H).
实施例7
(S)-3-(4-((4-((R)-1-(4-(环丙烷羰基)哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物11)
Figure PCTCN2021102873-appb-000025
第一步
(R)-(4-(1-(4-溴苯基)乙基)哌嗪-1-基)(环丙基)甲酮(11a)
将6b(200mg，0.75mmol)、环丙烷甲酰氯(93mg，0.89mmol)和二氯甲烷(5mL)混合，然后加入三乙胺(225mg，2.23mmol)。将反应混合物室温下搅拌过夜，然后在减压条件下浓缩至干。残余物用硅胶柱层析(石油醚/乙酸乙酯＝19/1至4/1)纯化，得到目标产物11a(200mg，80％)。
MS m/z(ESI):337[M+1]
第二步
(R)-4-(1-(4-(环丙烷羰基)哌嗪-1-基)乙基)苯甲酸甲酯(11b)
将11a(200mg，0.59mmol)、三乙胺(299mg，2.97mmol)和甲醇(20mL)混合，冷却到-40℃后用一氧化碳鼓泡30分钟，然后加入PdCl 2(dppf)(43mg，0.06mmol)。将混合物在封管中加热至100℃并搅拌18小时。冷却到室温后，减压除去溶剂，残余物用硅胶柱层析(石油醚/乙酸乙酯＝9/1至3/2)纯化，得到目标产物11b(120mg，75％)。
MS m/z(ESI):317[M+1]
第三步
(R)-环丙基(4-(1-(4-(羟甲基)苯基)乙基)哌嗪-1-基)甲酮(11c)
0℃下向11b(120mg，0.38mmol)的THF(5mL)溶液中加入LAH(22mg，0.57mmol)。搅拌1小时后，加入十水合硫酸钠(100mg)。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝1/1至0/100)纯化，得到目标产物11c(100mg，59％)。
MS m/z(ESI):289[M+1]
第四步
(R)-(4-(1-(4-(氯甲基)苯基)乙基)哌嗪-1-基)(环丙基)甲酮(11d)
向11c(100mg，0.35mmol)的二氯甲烷(5mL)溶液中添加氯化亚砜(0.5mL)。将反应混合物在室温下搅拌过夜，然后减压除去溶剂，得到目标产物11d(100mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):307[M+1]
第五步
(S)-5-氨基-4-(4-((4-((R)-1-(4-(环丙烷羰基)哌嗪-1-基)乙基)苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(11e)
将11d(100mg，粗品)、5f(101mg，0.30mmol)、碳酸钾(83mg，0.60mmol)和DMF(5mL)混合，加热到50℃并搅拌过夜。冷却到室温后，将混合物用水(10mL)稀释，用乙酸乙酯(3×20mL)萃取。合并的有机相经无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至19/1)纯化，得到目标产物11e(100mg，53％)。
MS m/z(ESI):605[M+1]
第六步
(S)-3-(4-((4-((R)-1-(4-(环丙烷羰基)哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(11)
将11e(100mg，0.17mmol)、TsOH·H 2O(66mg，0.35mmol)和乙腈(2mL)混合，加热到90℃下并搅拌过夜。冷却到室温后，减压除去溶剂，残余物用反相制备高效液相色谱纯化，得到目标产物11(10mg，固体，22％)。
MS m/z(ESI):531[M+1]
1H NMR(400MHz,CD 3OD)δ7.54–7.32(m,6H),7.29(d,J＝7.9Hz,1H),5.22(s,2H),5.13(dd,J＝13.4,5.2Hz,1H),4.45(q,J＝17.3Hz,2H),3.74(s,1H),3.57(s,1H),3.50–3.41(m,1H),3.01–2.83(m,2H),2.83–2.71(m,1H),2.62–2.42(m,4H),2.37(s,1H),2.22–2.10(m,1H),1.90(ddd,J＝12.8,7.9,4.7Hz,1H),1.40(d,J＝6.7Hz,3H),1.36–1.26(m,1H),0.87–0.73(m,4H).
实施例8
(S)-3-(4-((4-((R)-1-(4-环丙基哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物12)
Figure PCTCN2021102873-appb-000026
第一步
(R)-4-(1-(4-溴苯基)乙基)哌嗪-1-甲酸叔丁酯(12a)
将6b(对甲基苯磺酸盐，4g，9mmol)和THF(80mL)混合，然后加入三乙胺(3.7g，36mmol)。将混合物冷却至0℃，滴加二碳酸二叔丁酯(2.2g，9.9mmol)。室温搅拌17小时后，加入水(160mL)，然后用乙酸乙酯(2×100mL)萃取。将合并的有机相用无水硫酸钠干燥，过滤后，滤液浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至1/1)纯化，得到目标产物12a(3.07g，92％)。
MS m/z(ESI):369[M+1]
第二步
(R)-4-(1-(4-(甲氧基羰基)苯基)乙基)哌嗪-1-甲酸叔丁酯(12b)
在封管中将12a(3.07g，8.3mmol)溶于甲醇(120mL)中，加入三乙胺(4.21g，41.6mmol)。冷却至-40℃后，用一氧化碳鼓泡30分钟，然后加入PdCl 2(dppf)(1.21g，1.66mmol)。将混合物加热至100℃并搅拌18小时。冷却到室温后，垫硅藻土过滤。滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至1/1)纯化，得到目标产物12b(2.7g，93％)。
MS m/z(ESI):349[M+1]
第三步
(R)-4-(1-(哌嗪-1-基)乙基)苯甲酸甲酯(12c)
向12b(350mg，1.01mmol)的二氯甲烷(4mL)溶液中加入三氟乙酸(1mL)。室温下搅拌4小时，减压除去溶剂，得到目标产物12c(350mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):249[M+1]
第四步
(R)-4-(1-(4-(环丙基哌嗪-1-基)乙基)苯甲酸甲酯(12d)
将12c(140mg，0.56mmol)，(1-乙氧基环丙氧基)三甲基硅烷(295mg，1.69mmol)，乙酸(100mg，1.69mmol)和甲醇(5mL)的混合物中加入氰基硼氢化钠(106mg，1.69mmol)。将反应混合物在室温下搅拌过夜后，减压除去溶剂，残余物用硅胶柱层析(石油醚/乙酸乙酯＝9/1至7/3)纯化，得到目标产物12d(100mg，62％)。
MS m/z(ESI):289[M+1]
第五步
(R)-(4-(1-(4-(环丙基哌嗪-1-基)乙基)苯基)甲醇(12e)
0℃下向12d(100mg，0.35mmol)的THF(3mL)溶液中加入LAH(20mg，0.52mmol)。搅拌1小时后，加入十水合硫酸钠(100mg)。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝99/1至19/1)纯化，得到目标产物12e(75mg，83％)。
MS m/z(ESI):261[M+1]
第六步
(R)-1-(1-(4-(氯甲基)苯基)乙基)-4-环丙基哌嗪(12f)
向12e(80mg，0.31mmol)的二氯甲烷(5mL)溶液中加入氯化亚砜(1mL)。室温搅拌过夜后，减压除去溶剂，得到目标产物12f(80mg，粗品)，该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):279[M+1]
第七步
(S)-5-氨基-4-(4-((4-((R)-1-(4-环丙基哌嗪-1-基)乙基)苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(12g)
将12f(80mg，粗品)、5f(101mg，0.30mmol)、碳酸钾(83mg，0.60mmol)和DMF(5mL)混合，加热至50℃并搅拌过夜。冷却到室温后，用水(10mL)稀释混合物，然后用乙酸乙酯(3×20mL)萃取。有机相合并后，经无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，得到目标产物12g(120mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):577[M+1]
第八步
(S)-3-(4-((4-((R)-1-(4-环丙基哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮甲酸盐(12)
将12g(120mg，粗品)、TsOH·H 2O(95mg，0.50mmol)和乙腈(2mL)混合，加热至90℃并搅拌过夜。冷去到室温后，减压除去溶剂，残余物用反相制备高效液相色谱纯化，得到目标产物12(10.0mg，固体，6.5％)。
MS m/z(ESI):503[M+1]
1H NMR(400MHz,CD 3OD)δ8.45(s,1H),7.48(dd,J＝12.7,7.9Hz, 3H),7.40(d,J＝7.8Hz,3H),7.28(d,J＝8.0Hz,1H),5.24(s,2H),5.14(dd,J＝13.3,5.1Hz,1H),4.45(q,J＝17.3Hz,2H),3.79(dd,J＝13.4,6.6Hz,1H),2.98–2.71(m,8H),2.67(d,J＝15.8Hz,2H),2.56–2.45(m,1H),2.16(ddd,J＝10.3,5.2,2.9Hz,1H),1.85–1.76(m,1H),1.51(d,J＝6.8Hz,3H),0.50(dd,J＝12.1,5.7Hz,2H),0.48–0.38(m,2H).
实施例9
(S)-3-(4-((4-((R)-1-(4-(甲基磺酰基)哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物13)
Figure PCTCN2021102873-appb-000027
第一步
(R)-4-(1-(4-(甲基磺酰基)哌嗪-1-基)乙基)苯甲酸甲酯(13a)
将12c(140mg，0.56mmol)、甲基磺酰氯(97mg，0.85mmol)、三乙胺(90mg，0.90mmol)和二氯甲烷(5mL)混合，室温搅拌过夜后，减压除去溶剂，残余物用硅胶柱层析(石油醚/乙酸乙酯＝9/1至1/1)纯化，得到目标产物13a(100mg，54％)。
MS m/z(ESI):327[M+1]
第二步
(R)-(4-(1-(4-(甲基磺酰基)哌嗪-1-基)乙基)苯基)甲醇(13b)
0℃下向13a(100mg，0.31mmol)的THF(3mL)溶液加入LAH(18mg，0.46mmol)并搅拌1小时，然后加入十水合硫酸钠(100mg)。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝1％至5％)纯化，得到目标产物13b(80mg，88％)。
MS m/z(ESI):299[M+1]
第三步
(R)-1-(1-(4-(氯甲基)苯基)乙基)-4-(甲基磺酰基)哌嗪(13c)
向13b(40mg，0.13mmol)的二氯甲烷(5mL)溶液中加入氯化亚砜(1mL)。室温下搅拌过夜后，减压除去溶剂，得到目标产物13c(40mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):317[M+1]
第四步
(S)-5-氨基-4-(4-((4-((R)-1-(4-(甲基磺酰基)哌嗪-1-基)乙基)苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(13d)
将13c(40mg，粗品)、5f(101mg，0.30mmol)、碳酸钾(83mg，0.60mmol)和DMF(5mL)混合，加热到50℃并搅拌过夜。冷却到室温后，将混合物用水(20mL)稀释，然后用乙酸乙酯(3×20mL)萃取。合并的有机相经无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，得到目标产物13d(60mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):615[M+1]
第五步
(S)-3-(4-((4-((R)-1-(4-(甲基磺酰基)哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(13)
将13d(60mg，粗品)、TsOH·H 2O(95mg，0.50mmol)和乙腈(2mL)混合，加热到90℃并搅拌过夜。冷却到室温后，在减压条件下浓缩至干，残余物用反相制备高效液相色谱纯化，得到目标产物13(9.2mg，固体，13％)。
MS m/z(ESI):541[M+1]
1H NMR(400MHz,CD 3OD)δ7.52–7.33(m,6H),7.29(d,J＝8.0Hz,1H),5.21(s,2H),5.13(dd,J＝13.4,5.2Hz,1H),4.44(q,J＝17.4Hz,2H),3.50(q,J＝6.7Hz,1H),3.19(t,J＝4.9Hz,4H),2.97–2.84(m,1H),2.81(s,3H),2.76(ddd,J＝17.6,4.5,2.3Hz,1H),2.61(dt,J＝10.0,5.0Hz,2H),2.48(dd,J＝10.6,4.9Hz,3H),2.25–2.10(m,1H),1.39(d,J＝6.7Hz,3H).
实施例10
(S)-3-(4-((4-((R)-1-(4-甲基哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物14)
Figure PCTCN2021102873-appb-000028
第一步
(R)-1-(1-(4-(溴代苯基)乙基)-4-甲基哌嗪(14a)
将6b(200mg，0.75mmol)，甲醛水溶液(40％，0.3mL)，乙酸(122mg，2.0mmol)和甲醇(10mL)混合，然后加入氰基硼氢化钠(127mg，2.0mmol)。室温搅拌过夜后，减压除去溶剂，残余物用硅胶柱层析(石油醚/乙酸乙酯＝3/2至1/4)纯化，得到目标产物14a(200mg，95％)。
MS m/z(ESI):283[M+1]
第二步至第六步
(S)-3-(4-((4-((R)-1-(4-甲基哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(14)
参照化合物11中第二步至第六步的操作合成14，但在第二步中用14a代替11a。
MS m/z(ESI):283[M+1]
1H NMR(400MHz,CD 3OD)δ7.55–7.43(m,3H),7.38(t,J＝9.6Hz,3H),7.28(d,J＝7.7Hz,1H),5.23(s,2H),5.14(d,J＝8.6Hz,1H),4.44(q,J＝17.3Hz,2H),3.58(dd,J＝12.0,5.9Hz,1H),3.03(d,J＝27.6Hz,4H),2.96–2.73(m,4H),2.70(s,3H),2.64(s,2H),2.54–2.42(m,1H),2.17(dt,J＝8.5,6.3,1H),1.41(d,J＝6.3Hz,3H).
实施例11
(S)-3-(1-氧代-4-((4-((R)-1-(4-(嘧啶-4-基)哌嗪-1-基)乙基)苯甲基)氧基)异吲哚啉-2-基)哌啶-2,6-二酮(化合物15)
Figure PCTCN2021102873-appb-000029
第一步
(R)-4-(1-(4-(嘧啶-4-基)哌嗪-1-基)乙基)苯甲酸甲酯(15a)
将12c(200mg，0.81mmol)、4-氯嘧啶盐酸盐(243mg，1.61mmol)、碳酸钾(334mg，2.42mmol)和DMF(2mL)混合，加热到110℃并搅拌过夜。冷却到室温后，用水(10mL)稀释混合物，然后用乙酸乙酯(3×20mL)萃取。合并的有机相用无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝9/1 至1/1)纯化，得到目标产物15a(100mg，38％)。
MS m/z(ESI):327[M+1]
第二步
(R)-(4-(1-(4-(嘧啶-4-基)哌嗪-1-基)乙基)苯基)甲醇(15b)
在0℃下向15a(100mg，0.31mmol)的THF(5mL)溶液中加入LAH(18mg，0.46mmol)并搅拌1小时，然后加入十水合硫酸钠(100mg)。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝99/1至19/1)纯化，得到目标产物15b(40mg，44％)。
MS m/z(ESI):299[M+1]
第三步
(R)-4-(4-(1-(4-(氯甲基)苯基)乙基)哌嗪-1-基)嘧啶(15c)
向15b(40mg，0.13mmol)的二氯甲烷(4mL)溶液中加入氯化亚砜(1mL)。将反应混合物搅拌过夜后，减压浓缩至干，得到目标产物15c(40mg，粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):317[M+1]
第四步
(S)-5-氨基-5-氧代-4-(1-氧代-4-((4-((R)-1-(4-(嘧啶-4-基)哌嗪-1-基)乙基)苄基)氧基)异吲哚啉-2-基)戊酸叔丁酯(15d)
将15c(40mg，粗品)、5f(67mg，0.20mmol)、碳酸钾(56mg，0.40mmol)和DMF(1mL)混合，加热至50℃并搅拌过夜。冷却到室温后，将混合物用水(10mL)稀释，然后用乙酸乙酯(3×10mL)萃取。合并的有机相经无水硫酸钠干燥，过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝99/1至19/1)纯化，得到目标产物15d(40mg，两步49％)。
MS m/z(ESI):615[M+1]
第五步
(S)-3-(1-氧代-4-((4-((R)-1-(4-(嘧啶-4-基)哌嗪-1-基)乙基)苯甲基)氧基)异吲哚啉-2-基)哌啶-2,6-二酮(15)
将15d(40mg，0.07mmol)、TsOH·H 2O(37mg，0.20mmol)和乙腈(1mL)混合，加热到90℃并搅拌过夜。冷却到室温后，在减压条件下浓缩至干，残余物用反相制备高效液相色谱纯化，得到目标产物(20.0mg，固体，57％)。
MS m/z(ESI):541[M+1]
1H NMR(400MHz,CD 3OD)δ8.52(s,1H),8.14(d,J＝6.8Hz,1H),7.70(d,J＝8.2Hz,1H),7.50(t,J＝8.6Hz,2H),7.47–7.37(m,2H),7.29(d,J＝7.7Hz,1H),7.23(d,J＝7.9Hz,1H),6.88(d,J＝6.9Hz,1H),5.25(s,2H),5.14(dd,J＝13.3,5.2Hz,1H),4.45(q,J＝17.3Hz,2H),3.85(s,4H),3.00–2.66(m,6H),2.60-2.41(m,1H),2.36(s,1H),2.24–2.09(m,1H),1.53(d,J＝6.7Hz,3H).
化合物16和20参照化合物15中第一步至第五步的实验步骤合成，但在第一步中用不同的化合物代替4-氯嘧啶盐酸盐。
Figure PCTCN2021102873-appb-000030
化合物16和20的核磁数据如下：
Figure PCTCN2021102873-appb-000031
实施例12
(S)-3-(4-((4-((R)-1-(4-(2,4-二氟苯基)哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物17)
Figure PCTCN2021102873-appb-000032
第一步
(R)-4-(1-(4-(2,4-二氟苯基)哌嗪-1-基)乙基)苯甲酸甲酯(17a)
将12c(150mg，0.60mmol)、1-溴-2,4-二氟苯(232mg，1.20mmol)、叔丁醇钾(202mg，1.80mmol)、三(二亚苄基丙酮)二钯(35mg，0.06mmol)、XPhos(58mg，0.12mmol)和甲苯(2mL)混合，然后在氮气气氛下用微波反应器加热至130℃并搅拌1小时。冷却到室温后，减压除去溶剂，残余物用硅胶柱层析(石油醚/：10％至50％)纯化，得到目标产物17a(40mg，19％)。
MS m/z(ESI):361[M+1]
第二步至第五步
(S)-3-(4-((4-((R)-1-(4-(2,4-二氟苯基)哌嗪-1-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(17)
参照化合物15中第二步至第五步的操作合成17，但在第二步中用17a代替15a。
MS m/z(ESI):575[M+1]
1H NMR(400MHz,CD 3OD)δ7.54–7.44(m,3H),7.43–7.35(m,3H),7.30(d,J＝8.0Hz,1H),7.07–6.98(m,1H),6.93–6.82(m,2H),5.23(s,2H),5.13(dd,J＝13.4,5.2Hz,1H),4.45(q,J＝17.3Hz,2H),3.57(dd,J＝18.3,11.8Hz,1H),3.04(t,J＝4.7Hz,3H),2.95–2.84(m,1H),2.82–2.70(m,3H),2.66–2.58(m,2H),2.57–2.42(m,2H),2.21–2.12(m,1H),1.45(d,J＝6.7Hz,3H).
实施例13
(3S)-3-(4-((4-(1-(2-氧杂-5-氮杂双环[2.2.1]庚烷-5-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(化合物18和19)
Figure PCTCN2021102873-appb-000033
第一步
4-(1-(2-氧杂-5-氮杂双环[2.2.1]庚烷-5-基)乙基)苯甲酸甲酯(18a)
将4-(1-溴乙基)苯甲酸甲酯1e(243mg，1mmol)和2-氧杂-5-氮杂双环[2.2.1]庚烷盐酸盐(136mg，1mmol)溶于DMF(4mL)，然后加入碳酸钾(553mg，4mmol)。室温搅拌19小时后，用水(50mL)稀释，用乙酸乙酯(3×30mL)萃取。合并的有机相用水(3×30mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(石油醚/乙酸乙酯＝100/0至4/1)纯化，得到目标产物18a(212mg，81％)。
MS m/z(ESI):262[M+1]
第二步
(4-(1-(2-氧杂-5-氮杂双环[2.2.1]庚烷-5-基)乙基)苯基)甲醇(18b)
将18a(212mg，0.81mmol)溶于THF(5mL)，冷却至0℃，然后加入LAH(46mg，1.22mmol)。将混合物在0℃下搅拌30分钟，然后用NaOH水溶液(2.5N，1mL)淬灭并用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至19/1)纯化，得到目标产物18b(158mg，84％)。
MS m/z(ESI):234[M+1]
第三步
5-(1-(4-(氯甲基)苯基)乙基)-2-氧杂-5-氮杂双环[2.2.1]庚烷(18c)
向18b(158mg，0.68mmol)的二氯甲烷(5mL)溶液中加入氯化亚砜(161mg，1.35mmol)。将混合物在室温下搅拌2小时，然后减压除去溶剂，得到目标产物18c(粗品)。该产品未经进一步纯化，直接用于下一步反应。
MS m/z(ESI):252[M+1]
第四步
(4S)-4-(4-((4-(1-(2-氧杂-5-氮杂双环[2.2.1]庚烷-5-基)乙基)苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氨基-5-氧代戊酸叔丁酯(18d)
将18c(粗品，约0.68mmol)、5f(226mg，0.68mmol)、碳酸钾(376mg，2.72mmol)和DMF(5mL)混合，加热至50℃并搅拌48小时。冷却到室温后，用水(80mL)稀释，然后过滤收集沉淀物，得到目标产物18d(280mg，两步75％)。
MS m/z(ESI):550[M+1]
第五步
(3S)-3-(4-((4-(1-(2-氧杂-5-氮杂双环[2.2.1]庚烷-5-基)乙基)苯甲基)氧基)-1-氧代异吲哚啉-2-基)哌啶-2,6-二酮(18和19)
向18d(280mg，0.51mmol)的乙腈(30mL)溶液中加入TsOH·H 2O(194mg，1.02mmol)，加热到90℃并搅拌18小时。冷却到室温后，减压除去溶剂，然后加入饱和碳酸氢钠溶液(50mL)并用乙酸乙酯(2×50mL)萃取。合并的有机相用无水硫酸钠干燥，过滤后滤液在减压条件下浓缩至干，残余物用反相制备高效液相色谱纯化，得到目标产物18(异构体1，45mg，固体，19％)和19(异构体2，19mg，固体，8％)。
18的表征数据如下：
MS m/z(ESI):476[M+1]
1H NMR(400MHz,CD 3OD)δ7.51-7.36(m,6H),7.29(d,J＝8.0Hz,1H),5.21(s,2H),5.13(dd,J＝13.4,5.2Hz,1H),4.51-4.38(m,3H),4.13(d,J＝8.1Hz,1H),3.78(dd,J＝12.2,5.9Hz,2H),3.46(d,J＝8.1Hz,1H),3.09(dd,J＝10.4,1.5Hz,1H),2.95-2.84(m,1H),2.76(ddd,J＝17.6,4.6,2.4Hz,1H),2.60-2.43(m,2H),2.20-2.11(m,1H),1.88(d,J＝10.0Hz,1H),1.68(d,J＝10.0Hz,1H),1.38(d,J＝6.4Hz,3H).
19的表征数据如下：
MS m/z(ESI):476[M+1]
1H NMR(400MHz,CD 3OD)δ7.50-7.37(m,6H),7.30-7.26(m,1H),5.20(s,2H),5.13(dd,J＝13.3,5.2Hz,1H),4.43(dd,J＝30.0,19.2Hz,3H),4.07(d,J＝7.9Hz,1H),3.72-3.64(m,2H),3.60(dd,J＝7.9,1.8Hz,1H),2.94-2.84(m,1H),2.80-2.71(m,2H),2.54-2.43(m,2H),2.15(dtd,J＝12.7,5.2,2.3Hz,1H),1.92(dd,J＝9.4,2.3Hz,1H),1.73(d,J＝10.0Hz,1H),1.31(d,J＝6.5Hz,3H).
实施例14
5-(4-((R)-1-(4-(((2-((S)-2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氧基)甲基)苯基)乙基)哌嗪-1-基)-2-氰基吡啶(化合物21)
Figure PCTCN2021102873-appb-000034
第一步
(R)-4-(1-(4-(羟甲基)苯基)乙基)哌嗪-1-甲酸叔丁酯(21a)
0℃下向12b(2.7g，7.75mmol)的THF(30mL)溶液中加入LAH(353mg，9.30mmol)。0℃下搅拌1小时后，将混合物用水(0.5mL)淬灭，然后加入NaOH溶液(2.5N，4mL)。得到的混合物用THF(60mL)稀释后，用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，得到目标产物21a(2.75g，粗品)。其中1.97g粗品通过硅胶柱层析(石油醚/乙酸乙酯＝100/0至1/4)纯化，得到目标产物21a(1.75g)。
MS m/z(ESI):321[M+1]
第二步
(R)-(4-(1-(哌嗪-1-基)乙基)苯基)甲醇双盐酸盐(21b)
将21a(1.75g，5.46mmol)溶于1,4-二氧六环(20mL)，然后加入氯化氢的1,4-二氧六环溶液(4M，20mL)。室温搅拌18小时后，减压除去溶剂，残余物反相制备高效液相色谱纯化，得到目标产物21b(560mg，35％)。
MS m/z(ESI):221[M+1]
1H NMR(400MHz,DMSO-d 6)δ12.37(s,1H),9.76(s,1H),9.52(s,1H),7.60(d,J＝7.9Hz,2H),7.41(d,J＝8.0Hz,2H),4.60(s,1H),4.53(s,2H),4.18(s,2H),3.84(s,1H),3.69-3.29(m,4H),3.05(d,J＝51.8Hz,2H),1.71(d,J＝6.7Hz,3H).
第三步
(R)-5-(4-(1-(4-(羟甲基)苯基)乙基)哌嗪-1-基)-2-氰基吡啶(21c)
将21b(110mg，0.38mmol)、2-氰基-5-氟吡啶(51mg，0.42mmol)、碳酸铯(371mg，1.14mmol)和DMF(4mL)混合，加热至60℃并搅拌2小时。冷却到室温后，反应混合物通过硅藻土过滤。滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至19/1)纯化，得到目标产物21c(24mg，52％)。
MS m/z(ESI):323[M+1]
第四步
(S)-5-氨基-4-(4-((4-((R)-1-(4-(6-氰基吡啶-3-基)哌嗪-1-基)乙基)苄基)氧基)-1-氧代异吲哚啉-2-基)-5-氧代戊酸叔丁酯(21d)
将21c(64mg，0.20mmol)溶于二氯甲烷(10mL)，然后加入氯化亚砜(48mg，0.40mmol)。室温搅拌2小时后，在减压条件下浓缩至干。将残余物溶于DMF(4mL)中，然后加入5f(67mg，0.20mmol)和碳酸钾(138mg，1mmol)。将混合物加热至50℃并搅拌20小时。冷却到室温后过滤，滤液在减压条件下浓缩至干，残余物用硅胶柱层析(二氯甲烷/甲醇＝100/0至19/1)纯化，得到目标产物21d(92mg，72％)。
MS m/z(ESI):639[M+1]
第五步
5-(4-((R)-1-(4-(((2-((S)-2,6-二氧代哌啶-3-基)-1-氧代异吲哚啉-4-基)氧基)甲基)苯基)乙基)哌嗪-1-基)-2-氰基吡啶甲酸盐(21)
将21d(156mg，0.29mmol)、TsOH·H 2O(110mg，0.58mmol)和乙腈(10mL)混合，加热至90℃并搅拌20小时。冷却至室温后，用饱和碳酸氢钠溶液调节至pH＝8，然后用乙酸乙酯(3×50mL)萃取。合并的有机相依次用饱和碳酸氢钠溶液(50mL)和饱和食盐水(50mL)洗涤，然后用无水硫酸钠干燥。过滤后，滤液在减压条件下浓缩至干，残余物用反相制备高校液相色谱纯化，得到目标产物22(62mg，固体，含0.65当量甲酸，45％)。
MS m/z(ESI):565[M+1]
1H NMR(400MHz,DMSO-d 6)δ10.95(s,1H),8.37(d,J＝2.9Hz,1H),8.16(s,0.65H),7.72(d,J＝8.9Hz,1H),7.48(dd,J＝16.8,8.1Hz,3H),7.38–7.28(m,5H),5.23(s,2H),5.11(dd,J＝13.3,5.1Hz,1H),4.42(d,J＝17.4Hz,1H),4.26(d,J＝17.5Hz,1H),3.48(q,J＝6.6Hz,1H),3.37(t,J＝4.9Hz,4H),2.96-2.85(m,1H),2.56(dd,J＝15.0,10.7Hz,3H),2.48-2.38(m,3H),1.98(dt,J＝10.2,5.0Hz,1H),1.33(d,J＝6.7Hz,3H).
化合物22参照化合物21中第一步至第五步的实验步骤合成，但在第三步中用6-氯-5-氟烟腈代替2-氰基-5-氟吡啶。
Figure PCTCN2021102873-appb-000035
化合物22的核磁数据如下：
Figure PCTCN2021102873-appb-000036
生物学实验
实施例15
NCI-H929细胞增殖抑制的测定
使用发光细胞活力测试实验评估本发明的化合物对NCI-H929人骨髓瘤细胞增殖的影响。
实验方法概述如下：
将化合物在DMSO中溶解并稀释至5mM，然后用DMSO进行4倍的系列稀释至最低浓度为0.31μM，每个浓度点再用RPMI 1640培养基(Thermo Fisher，货号为72400-047)稀释50倍。如果化合物IC 50值较低，可降低化合物的起始浓度。
NCI-H929(南京科佰，货号为CBP60243)细胞被培养在RPMI 1640完全培养基[其中含10％FBS(GBICO，货号为10099-141)和100units/mL青链霉素混合液(Thermo Fisher，货号为15140122)]中。将细胞(15000个细胞/mL)接种于96孔板的90μL完全培养基中，培养过夜后每孔加入10μL化合物溶液，于37℃、5％CO 2的培养箱中培养6天，然后按CellTilter-Glo(CTG)试剂盒(Promega，货号为G7572)说明书操作，将细胞培养板取出平衡至室温，加50μL CTG试剂充分裂解，室温放置10分钟后用酶标仪(EnVision，Perkin Elmer)读取冷光信号。以含0.2％DMSO培养基组作为0％抑制，使用XLfit软件绘制化合物抑制曲线并计算其抑制的IC 50，实验结果见表1。
实施例16
影响人外周血单个核细胞的细胞中TNFα、IL-2分泌的测定
通过酶联免疫吸附(ELISA)的方法检测本发明的化合物对人外周血单个核细胞的细胞(hPBMC)中TNFα和IL-2分泌量的影响，从而评估其免疫调节功能。
实验方法概述如下：
用EDTA抗凝管采集健康自愿者的外周血，用含2％胎牛血清(Gibco，货号为10099-141)的磷酸盐缓冲液(PBS)按等体积稀释后，转移30mL至预先加入15mL密度梯度离心液(Sigma，货号为10771)的Sepmate-50离心管(Stemcell，货号为86450)中，室温下1200×g离心10分钟后，上层含PBMC的液体转移至新的50mL离心管，室温下300×g离心8分钟，弃上清，将PBMC用RPMI 1640培养基(Gibco，货号为72400-047)重悬至5×10 6/mL，在96孔板中每孔加入80μL。
将化合物在DMSO中溶解并稀释至5mM(如果化合物IC 50值较低，可降低此起始浓度)，然后用DMSO进行4倍的系列稀释至最低浓度为0.31μM，每个浓度点再用RPMI 1640培养基稀释50倍，取10μL加入上述96孔板的细胞中。将该96孔板置于37℃、5％CO 2的培养箱中培养1小时后，加入10μL浓度为100ng/mL的LPS(Sigma，货号L-2880)，于37℃、5％CO 2的培养箱中继续培养过夜。收集上清，用于TNFα的检测。
也可将上述96孔板置于37℃、5％CO 2的培养箱中培养1小时后，加入10μL浓度为500ng/mL的anti-CD3抗体(Thermo Fisher，货号为14-0037-82)，于37℃、5％CO 2的培养箱中继续培养72小时。收集上清，用于IL-2的检测。
TNFα和IL-2的检测分别按照各自的ELISA试剂盒(R&D，货号分别为DY210和DY202)说明书进行，得到各孔的OD450值。以含0.2％DMSO培养基组作为0％抑制或刺激，使用XLfit软件绘制化合物抑制或刺激曲线并计算相应的IC 50或EC 50，实验结果见表1。
表1
Figure PCTCN2021102873-appb-000037
Figure PCTCN2021102873-appb-000038
实施例17
hERG钾离子通道阻断的测定
通过测定对hERG钾离子通道阻断作用评估本发明的化合物对可能心律失常的影响。
实验方法概述如下：
细胞外液：140mM NaCl、3.5mM KCl、1mM MgCl 2、2mM CaCl 2、10mM D-glucose、10mM HEPES、1.25mM NaH 2PO 4、pH＝7.4。
电极内液：20mM KCl、115mM K-aspartate、1mM MgCl 2、5mM EGTA、10mM HEPES、2mM Na 2-ATP、pH＝7.2。
化合物溶液：将待测化合物在DMSO中溶解并配成10mΜ的储液，再用DMSO将其稀释为3mM，接着用细胞外液稀释成3μΜ的溶液，以备后续使用。
细胞培养：hERG钾通道稳定过表达的HEK293细胞系(Creacell，货号为A-0320)在含有10％胎牛血清(Gibco，货号为1428478)和0.8mg/mL G418(Amresco，货号为E859-5G)的DMEM培养基(Gibco，货号为11995-065)中培养，培养温度为37℃，二氧化碳浓度为5％。除去旧培养基并用PBS(Gibco，货号为1009-141)洗一次，然后加入1mL TrypLE TM Express溶液(Gibco，货号为12604021)，37℃孵育30秒。当细胞从培养皿底脱离，加入5mL 37℃预热的完全培养基，转移细胞悬液至无菌的离心管中，于1000rpm离心5分钟后收集细胞。将细胞接种于6cm细胞培养皿，每个细胞培养皿接种细胞量为2.5*10 5(最终体积：5mL)。膜片钳检测实验之前将3*10 3个细胞铺到盖玻片上，在24孔板中培养(最终体积：500μL)，18个小时后进行检测。
全细胞膜片钳记录全细胞hERG钾电流的电压刺激方案：当形成全细胞封接后细胞膜电压钳制于-80mV。钳制电压由-80mV除极至-50mV并维持0.5秒(作为漏电流检测)，然后阶跃至30mV并维持2.5秒，再迅速恢复至-50mV并维持4秒以激发出hERG通道的尾电流(Peak tail current)，每隔10秒记录1次hERG钾电流。实验数据由 EPC-10放大器(HEKA)进行采集并储存于PatchMaster(HEKA v2x73)软件中。
测定：用微电极拉制仪(Sutter Instruments)将毛细玻璃管(Sutter Instruments)拉制成记录电极。将载有细胞的盖玻片从放置于培养箱的24孔板中取出，然后置于倒置显微镜下。记录电极内灌注电极内液，然后操纵微电极操纵仪(Sutter Instruments)，将记录电极接触到细胞表面，给予负压抽吸，形成GΩ封接；接着进行快速电容补偿；然后继续给予负压抽吸，直至吸破细胞膜，形成全细胞记录模式。在全细胞记录模式下，进行慢速电容的补偿并记录膜电容及串联电阻，期间不给予漏电补偿。当全细胞记录的hERG尾电流稳定3-5分钟后，将8mL不含化合物的细胞外液(空白对照)和8mL 3μM待测化合物溶液利用重力灌流的方法，依次流经记录浴槽从而作用于细胞5分钟(或者至电流稳定)。每一个细胞在不含化合物的细胞外液中检测到的电流作为自己的对照组。独立重复检测2-3个细胞。所有电生理实验在室温下进行。
数据分析：首先将待测化合物作用后的电流和空白对照电流标准化
Figure PCTCN2021102873-appb-000039
然后计算与这个化合物浓度对应的抑制率
Figure PCTCN2021102873-appb-000040
结果见表2.
表2
Figure PCTCN2021102873-appb-000041
实施例18
大鼠体内药代动力学实验
将待测化合物溶解在5％DMA+20％Solutol+75％Saline溶媒中，配制成0.5mg/mL的给药溶液。
在1mg/kg剂量下以静脉注射方式(IV)分别给予3只进食雄性Sprague-Dawley大鼠2mL/kg的给药溶液，在给药后0.083、0.25、0.5、1、2、4、8和24小时采集血样。在5mg/kg剂量下以灌胃方式(PO)分别给予另3只进食雄性Sprague-Dawley大鼠10mL/kg的给药溶液，在给药后0.25、0.5、1、2、4、8和24小时采集血样。
血浆中待测化合物的浓度用API-4500质谱仪进行LC-MS/MS定量分析，血浆定量限(LOQ)为1ng/mL。用WinNonlin计算药代动力学 (PK)参数，结果汇总在表3。
表3
Figure PCTCN2021102873-appb-000042

Claims (13)

  1. 一种通式(I)所示的化合物，或其可药用的盐、稳定同位素衍生物、异构体：
    Figure PCTCN2021102873-appb-100001
    其中：
    环A为含N的4-10元杂环；
    R 1为D或卤素；
    R 2和R 3各自独立地选自H、D、卤素、氰基、C 1-6烷基、C 3-6环烷基或3-8元杂环基，但两者不可同时为H和/或D，或者R 2和R 3与其相连的碳原子共同组成C 3-6环烷烃或3-8元杂环；
    R 4为D、卤素、氰基、氧代、C 1-6烷基、C 3-6环烷基、-OC 1-6烷基、-C(O)C 1-6烷基、-C(O)C 3-6环烷基、-S(O) 2C 1-6烷基、-S(O) 2C 3-6环烷基、芳基或杂芳基，其中所述烷基、环烷基、芳基和杂芳基的一个或多个氢任选被D、卤素、氰基、C 1-2烷基或氟代C 1-2烷基所取代；
    m为整数0-4；且
    n为整数0-2。
  2. 根据权利要求1所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，其中环A为含一个N原子的4-10元单环杂环，或稠合、桥接或螺双环杂环(比如吗啉、哌啶、硫代吗啉-1,1-二氧化物、2-氧杂-5-氮双环[2.2.1]庚烷等)；R 4为D、卤素、氧代、-CF 3或C 1-6烷基。
  3. 根据权利要求1所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，其中环A为含两个N原子的6-10元单杂环，或稠合、桥接或螺双环杂环(比如哌嗪、3,6-二氮杂二环[3.1.1]庚烷、2,6-二氮杂螺[3.3]庚烷等)，优选环A为哌嗪；R 4为与第二个N原子相连的C 1-6烷基、C 3-6环烷基、-C(O)C 1-6烷基、-C(O)C 3-6环烷基、-S(O) 2C 1-6烷基、-S(O) 2C 3-6环烷基、苯基或5-6元含N、O和/或S的杂芳基，其中所述烷基、环烷基、苯基和杂芳基的一个或多个氢任选被D、卤素、氰基、C 1- 2烷基或氟代C 1-2烷基所取代；n为0或1。
  4. 根据权利要求1-3任一项所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，其中R 2为H；R 3为氰基、C 1-6烷基或C 3-6环烷基，优选R 3为甲基或氰基。
  5. 根据权利要求1-3任一项所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，其中R 2和R 3都为甲基。
  6. 根据权利要求1-3任一项所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，其中R 2和R 3与其相连的碳原子共同组成C 3-6环烷烃。
  7. 根据前述权利要求任一项所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，其中m为0。
  8. 根据权利要求1所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，其为以下通式(II)的化合物：
    Figure PCTCN2021102873-appb-100002
    其中：
    R 5为H、C 1-6烷基、C 3-6环烷基、-C(O)C 1-6烷基、-C(O)C 3-6环烷基、-S(O) 2C 1-6烷基、苯基或5-6元含N、O和/或S的杂芳基，其中所述烷基、环烷基、苯基和杂芳基的一个或多个氢任选被D、卤素、氰基或C 1- 2烷基所取代，优选R 5为苯基、吡啶基或嘧啶基，其中所述苯基、吡啶基和嘧啶基的一个或两个氢任选被F或氰基所取代。
  9. 根据权利要求1所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，所述化合物选自：
    Figure PCTCN2021102873-appb-100003
    Figure PCTCN2021102873-appb-100004
    Figure PCTCN2021102873-appb-100005
  10. 权利要求9所述的化合物为：
    Figure PCTCN2021102873-appb-100006
  11. 药物组合物，其中包含权利要求1-10任一项所述的化合物或其可药用的盐、稳定同位素衍生物、异构体和药学上可接受的载体或辅料。
  12. 药物组合物，其中包含权利要求1-10任一项所述的化合物或其可药用的盐、稳定同位素衍生物、异构体和至少一种额外的药物，其中所述至少一种额外的药物为化学治疗剂、免疫和/或炎症调节剂、神经相关疾病调节剂等。
  13. 一种治疗或者预防Aiolos、Ikaros、Helios、CK1α、GSPT1、IL-2、IL-6、TNFα、IFNγ、VEGF等介导的相关疾病的方法，所述方法包括给予有需要的患者治疗有效量的权利要求1-12任一项所述的化合物或其可药用的盐、稳定同位素衍生物、异构体，或包含所述化合物的药物组合物，所述疾病包括但不限于血液瘤、实体瘤、自身免疫性疾病、炎症、神经退行性疾病、皮肤疾病等。
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