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WO2025011623A1 - Cyanoquinoline-targeting protein degradation molecule, preparation method therefor and use thereof - Google Patents

Cyanoquinoline-targeting protein degradation molecule, preparation method therefor and use thereof Download PDF

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Publication number
WO2025011623A1
WO2025011623A1 PCT/CN2024/105021 CN2024105021W WO2025011623A1 WO 2025011623 A1 WO2025011623 A1 WO 2025011623A1 CN 2024105021 W CN2024105021 W CN 2024105021W WO 2025011623 A1 WO2025011623 A1 WO 2025011623A1
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mmol
added
compound
room temperature
methyl
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French (fr)
Chinese (zh)
Inventor
叶斌
唐悦
王新元
金健
王飞
于军
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Shanghai Yidi Biotechnology Co Ltd
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Shanghai Yidi Biotechnology Co Ltd
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    • 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
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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

Definitions

  • the present invention belongs to the field of medicine, and in particular, relates to cyanoquinoline targeted protein degradation molecules, and preparation methods and applications thereof.
  • Prostate cancer is a common male malignancy, second only to lung cancer in incidence, and is the second leading cause of death among adult men worldwide.
  • androgen deprivation therapy ADT is the main treatment for early prostate cancer, including surgical castration and medical castration.
  • ADT cannot cure prostate cancer.
  • CRPC castration-resistant prostate cancer
  • patients are no longer sensitive to ADT, with a median survival of less than 20 months.
  • AR androgen receptor
  • representative androgen receptor inhibitors for the treatment of CRPC include abiraterone, enzalutamide, etc.
  • 15-25% of patients do not respond to second-generation hormone therapy such as abiraterone and enzalutamide, and most responsive patients will eventually develop severe drug resistance, leading to poor prognosis.
  • UPS The ubiquitin-proteasome system
  • UPS The ubiquitin-proteasome system
  • Ub The ubiquitin-proteasome system
  • E1 ubiquitin activating enzyme
  • E2 ubiquitin conjugating enzyme
  • E3 ubiquitin-protein ligase
  • 26S proteasome The ubiquitin activating enzyme system is responsible for activating ubiquitin and binding it to the protein to be degraded to form a polyubiquitin chain of the target protein, i.e. ubiquitination.
  • the proteasome system can recognize ubiquitinated proteins and degrade them.
  • Protein proteolysis-targeting chimeras are hybrid bifunctional small molecule compounds, including a small molecule compound that can bind to a target protein, a linker group at a suitable position, and a small molecule compound that can bind to an E3 ubiquitin ligase.
  • PROTACs work by bringing the target protein and the E3 in the cell together to form a target protein-PROTACs-E3 ternary polymer, adding a ubiquitinated protein tag to the target protein through the E3 ubiquitin ligase, and using the ubiquitin-proteasome system (UPS) to specifically degrade the target protein.
  • UPS ubiquitin-proteasome system
  • the present invention provides a compound represented by general formula (I), its stereoisomers or pharmaceutically acceptable salts thereof:
  • M 1 is selected from N or CR 1 ;
  • M 2 is selected from N or CR 2 ;
  • M 3 is selected from N or CR 3 ;
  • M 4 is selected from N or CR 4 ;
  • M 5 is selected from N or CR 5 ;
  • R 1 , R 2 , R 3 , R 4 and R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, hydroxyalkoxy, cycloalkyl or heterocyclyl;
  • Ring A, Ring B and Ring C are each independently selected from cycloalkyl, heterocyclyl, aryl or heteroaryl;
  • L2 is -Ak1-Cy1-Ak2-Cy2-Ak3-
  • Cy1 and Cy2 are each independently selected from a bond, a cyclohexylene, a piperidinylene, a piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene or a cyclopentylene, and the cyclohexylene, piperidinylene, piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene and a cyclopentylene may optionally be further substituted; or, any two atoms on the cyclohexylene, piperidinylene, piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene and a cyclopentylene may be linked to form a cycloalkyl or heterocyclic group;
  • Ra , Rb and Rc are each independently selected from hydrogen, deuterium, halogen, oxo, cyano, hydroxy, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, and the alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl may be further substituted;
  • any two R c are linked to form a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, and the cycloalkyl group, the heterocyclic group, the aryl group and the heteroaryl group may be further substituted;
  • R a and L 1 are linked to form a cycloalkyl group, a heterocyclic group or a heteroaryl group, and the cycloalkyl group, the heterocyclic group and the heteroaryl group may be further substituted;
  • x, y and z are each independently selected from 0, 1, 2 or 3;
  • n1, n2, n3, n4, n5, n6, n7, n8, n9, n10, n11, n12, n13, n14, n15, n16 and n17 are independent Selected from 0, 1, 2 or 3.
  • R d is selected from hydrogen, halogen or C 1-3 alkyl
  • p1 and p2 are each independently selected from 0, 1, 2, 3 or 4
  • L 4 is selected from a bond, -CH 2 -, -NH-, -O-, -S- or -C(O)-.
  • M 2 , M 3 , M 4 and M 5 are not N at the same time.
  • the ring A mentioned above is selected from phenyl, 5-6 membered monocyclic heteroaryl or 9-10 membered bicyclic heterocyclic group;
  • ring A is selected from phenyl, pyrazolyl, pyrrolyl, pyridyl, pyrimidinyl, benzopyridazinone, benzopyrazolyl or benzopyrrolidinyl;
  • Ring B is selected from cyclohexyl or cyclobutane
  • ring C is selected from phenyl, 5-6 membered monocyclic heteroaryl, 9-10 membered bicyclic heterocyclyl, 9-10 membered bicyclic heteroaryl;
  • ring C is selected from benzopyridyl, phenyl, benzothiazolyl, benzopyrazinyl, pyridopyrazolyl, pyridotriazolyl, pyridophenyl, benzothiadiazole or benzopyridazinyl.
  • the ring A is selected from phenyl, pyridyl,
  • Ra , Rb and Rc described above are each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, oxo, C1-3 alkyl, C1-3 alkoxy, hydroxyC1-3 alkyl, hydroxyC1-3 alkoxy, C1-3 haloalkyl, C1-3 haloalkoxy, C3-4 cycloalkyl or 3-4 membered heterocyclic group.
  • R 1 , R 2 , R 3 , R 4 and R 5 mentioned above are each independently selected from hydrogen, deuterium, halogen, cyano, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, hydroxy C 1-3 alkyl, hydroxy C 1-3 alkoxy, C 3-4 cycloalkyl or 3-4 membered heterocyclic group.
  • L 1 mentioned above is selected from a bond, -C(O)NH-, -NH-, -O-, -NHC(O)-, -C(O)-, -CH 2 - or -(CH 2 ) 2 -;
  • L 2 is selected from
  • L 3 and L 4 are each independently selected from a bond, -CH 2 -, -NH-, -O-, -S- or -C(O)-;
  • M 6 and M 7 are each independently selected from -CR 6 R 7 - or -C(O)-;
  • M 8 is selected from N or CR 8 ;
  • R 6 , R 7 and R 8 are each independently selected from hydrogen, deuterium, fluorine, chlorine, hydroxyl, cyano, C 1-3 alkyl, hydroxy C 1-3 alkyl, C 1-3 alkoxy, hydroxy C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, C 3-4 cycloalkyl or 3-4 membered heterocyclyl;
  • j, p1 and p2 are each independently selected from 0, 1, 2, 3 or 4.
  • Ra described above in the present invention is selected from hydrogen, fluorine, chlorine, methyl or ethyl.
  • the R b mentioned above in the present invention is independently selected from hydrogen, methyl, ethyl, methoxy, ethoxy, hydroxymethyl or hydroxyethyl.
  • R c mentioned above in the present invention is independently selected from hydrogen, cyano, fluorine, chlorine, hydroxyl, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, hydroxymethyl or hydroxyethyl.
  • R d as described above in the present invention is selected from hydrogen, fluorine, chlorine, methyl or ethyl.
  • R 2 , R 3 , R 4 and R 5 described above are each independently selected from hydrogen, fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, hydroxymethyl or hydroxyethyl.
  • the above-mentioned compound is selected from the following compounds in Table 5:
  • the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned compound, its stereoisomer or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.
  • the present invention also relates to a preferred embodiment, involving the use of each compound of the general formula, its stereoisomers or pharmaceutically acceptable salts, or the composition described above in the preparation of drugs for androgen receptor-regulated related diseases.
  • the above-mentioned disease is selected from prostate cancer, breast cancer, and Kennedy's disease.
  • the present invention also provides a drug for treating diseases regulated by androgen receptors, wherein the drug is a preparation prepared with the above-mentioned compound, or its isotope compound, or its optical isomer, or its tautomer, or its pharmaceutically acceptable salt, or its prodrug, or its solvate as an active ingredient and a pharmaceutically acceptable excipient.
  • the present invention also provides a compound represented by formula (A-1), a stereoisomer thereof or a pharmaceutically acceptable salt thereof:
  • R 10 is selected from hydrogen or an amino protecting group
  • Ring A, M 1 , L 1 , L 4 , Ra , p1, p2 and x are as defined above in the present invention.
  • the amino protecting group is preferably SEM, tert-butyloxycarbonyl, -CH2 -tert-butyloxycarbonyl, -CH2COOH , -COCH3 , ethoxycarbonyl, benzyl, p-methoxybenzyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl or allyloxycarbonyl.
  • the present invention also relates to the use of the structural unit of formula (A-1-1) as a structural unit of a protein degradation inhibitor:
  • ring A, M 1 , L 1 , L 4 , Ra , p1, p2 and x are as defined above in the present invention.
  • the compound of formula (A-1) is further represented by formula (A-1-a):
  • formula (A-1-1) is further shown as formula (A-1-1-a):
  • the compound of formula (I) provided by the present invention can target androgen receptors in prostate cancer cells and inhibit the proliferation of prostate cancer cells, while also showing good metabolic stability and pharmacokinetic properties.
  • the compound of the present invention has good application prospects in the preparation of protein degradation targeted chimeras of androgen receptors, and in the preparation of drugs for treating related diseases regulated by androgen receptors (including prostate cancer, breast cancer, and Kennedy's disease).
  • alkyl refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms.
  • Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc., and various branched isomers thereof, etc.
  • Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl, and hydroxy-substituted alkyl are preferred in the present invention.
  • alkylene refers to an alkyl group in which one hydrogen atom is further substituted.
  • methylene refers to -CH2-
  • ethylene refers to -( CH2 ) 2-
  • propylene refers to -( CH2 ) 3-
  • butylene refers to -( CH2 ) 4- , and the like.
  • alkenyl refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc.
  • Alkenyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.
  • alkenylene refers to an alkenyl group in which one of its hydrogen atoms is further substituted, for example: “ethenylene” refers to -(CH) 2 -.
  • cycloalkyl refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms, and further preferably 3 to 4 carbon atoms.
  • Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include spirocyclic, fused and bridged cycloalkyls, non-limiting examples of which include The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, etc.
  • the cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.
  • cycloalkylene refers to a cycloalkyl group in which one hydrogen atom is further substituted.
  • Non-limiting examples include: cyclohexylene,
  • the cycloalkylene group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.
  • it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 12 ring atoms; most preferably, it contains 3 to 6 ring atoms, and further preferably, it contains 3 to 4 ring atoms.
  • Non-limiting examples of monocyclic heterocyclic groups include oxetane, azetane, tetrahydropyranyl, azepanyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably oxetane, thietane, azetane, tetrahydrofuranyl, tetrahydropyranyl, 1-aminoylidene-1-oxothiopyran, azepanyl, piperidinyl and piperazinyl.
  • Polycyclic heterocyclic groups include spirocyclic, condensed ring and bridged ring heterocyclic groups, non-limiting examples include
  • the heterocyclic group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.
  • heterocyclylene refers to a heterocyclyl group in which one hydrogen atom is further substituted, and non-limiting examples thereof include: piperidinylene, piperazinylene, pyrrolopyrrolylene, diazaspiro[5.5]undecylene, azaspiro[5.5]undecylene, benzopiperidinylene, azetidinylene, diazetidinylene, pyrrolidinylene, azaspiro[3.5]nonanylene, diazaspiro[3.5]nonanylene, azabicyclo[3.1.1]heptanylene, azaspiro[2.5]octanylene,
  • the heterocyclylene group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyan
  • aryl refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated ⁇ electron system, preferably 6- to 10-membered, more preferably phenyl.
  • the aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein The ring connected to the parent structure is an aryl ring.
  • the aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.
  • arylene refers to an aryl group in which one hydrogen atom is further substituted, and non-limiting examples thereof include:
  • the arylene group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.
  • heteroaryl refers to a heteroaromatic system containing 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen.
  • Heteroaryl is preferably a 5-8-membered monoheteroaryl or an 8-14-membered diheteroaryl, more preferably a 5-membered monoheteroaryl, a 6-membered monoheteroaryl or a 9-membered diheteroaryl, such as imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, pyridoimidazolyl, pyrimidoimidazolyl, etc., preferably pyridoimidazolyl and pyrimidoimidazo
  • the heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.
  • heteroarylene refers to a heteroaryl group in which one hydrogen atom is further substituted, and non-limiting examples thereof include pyridylene, pyrimidylene, pyrazolylene, pyridazinylene, pyrazinylene,
  • alkoxy refers to-O-(alkyl) and-O-(non-substituted cycloalkyl), wherein the definition of alkyl is as described above.
  • the non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy.
  • Alkoxy can be optionally substituted or non-substituted, and when substituted, substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.
  • substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, hetero
  • Haloalkyl refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above. Non-limiting examples include: trifluoromethyl, difluoromethyl.
  • Haloalkoxy refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.
  • Hydroalkyl refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above. Non-limiting examples include: -C(CH 3 ) 2 (OH).
  • Hydroalkoxy means an alkoxy group substituted with a hydroxy group wherein alkoxy is as defined above.
  • the chiral carbon in the compounds of the present invention is in R configuration or S configuration.
  • the hydrogen atoms described in the present invention may be replaced by their isotope deuterium atoms, and any hydrogen atoms in the example compounds of the present invention may also be replaced by deuterium atoms.
  • Optional or “optionally” means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur.
  • a heterocyclic group optionally substituted with an alkyl group means that an alkyl group may but need not be present, and the description includes 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, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (e.g. olefinic) bonds.
  • substituents in "optionally may be further substituted” are each independently selected from deuterium, halogen, cyano, hydroxy, nitro, amino, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein, halogen, cyano, hydroxy, nitro, amino, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are as defined above.
  • “Pharmaceutical composition” means a mixture containing one or more compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients.
  • the purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.
  • “Pharmaceutically acceptable salts” refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.
  • PG in the compound of formula III represents a protecting group, such as tert-butyl or methyl.
  • the compound of formula III undergoes an aromatic nucleophilic substitution reaction with a compound of formula IV-A or IV-B under alkaline conditions (such as using N,N-diisopropylethylamine as a base) in a suitable solvent such as N,N-dimethylformamide, or undergoes a Buchwald-Hartwig coupling reaction using a palladium catalyst such as [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride in a suitable solvent such as toluene and using an appropriate base such as cesium carbonate to obtain a compound of formula V-A or V-B.
  • alkaline conditions such as using N,N-diisopropylethylamine as a base
  • a suitable solvent such as N,N-dimethylformamide
  • a palladium catalyst such as [1,1'-bis
  • the deprotected product of the compound of formula VA or VB, the compound of formula VI-A or VI-B, is reacted with the compound of formula VII using conventional methods reported in the literature, such as using N,N-diisopropylethylamine as a base and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) or other condensation reagents as an activating reagent to obtain an amide compound of formula VIII-A or VIII-B.
  • HATU 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate
  • the compound of formula VIII-A is oxidized (e.g., using Dess-Martin reagent as an oxidant) to obtain a compound of formula IX; the compound of formula VIII-B is hydrolyzed under acidic conditions (e.g., hydrochloric acid) to obtain a compound of formula IX.
  • the compound of formula IX is subjected to a reductive amination reaction with a compound of formula X in the presence of a suitable reducing agent (e.g., sodium acetate borohydride) to obtain a final product, a compound of formula II.
  • a suitable reducing agent e.g., sodium acetate borohydride
  • the compound of formula VIII-A can be converted into a sulfonate compound of formula XI by conventional methods reported in literature, such as using triethylamine as a base and methylsulfonyl chloride.
  • the compound of formula XI undergoes a nucleophilic substitution reaction with the compound of formula X in a suitable solvent such as N,N-dimethylformamide under alkaline conditions (such as using potassium carbonate as a base) to obtain the final product, the compound of formula II.
  • Another route is to oxidize the compound of formula VA (such as using Dess-Martin reagent as an oxidant) to obtain a compound of formula XII; the compound of formula VB is hydrolyzed under acidic conditions (such as hydrochloric acid) to obtain a compound of formula XII.
  • the compound of formula XII and the compound of formula X are subjected to a reductive amination reaction under a suitable reducing agent (such as sodium acetate borohydride) to obtain a compound of formula XIV.
  • a suitable reducing agent such as sodium acetate borohydride
  • the compound of formula VA is converted into a sulfonate compound of formula XIII using conventional methods reported in the literature, such as using triethylamine as a base and methylsulfonyl chloride.
  • Formula XIII The compound of formula X undergoes nucleophilic substitution reaction with the compound of formula X in a suitable solvent such as N,N-dimethylformamide under alkaline conditions (such as using potassium carbonate as a base) to obtain a compound of formula XIV.
  • the deprotected product of the compound of formula XIV, the compound of formula XV, is reacted with the compound of formula VII using conventional methods reported in the literature, such as using N,N-diisopropylethylamine as a base, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) or other condensation reagents to obtain the final product, the compound of formula II.
  • HATU 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate
  • the starting materials and reagents used in the examples of the present invention are known and can be purchased on the market, or can be synthesized by or according to methods known in the art.
  • tert-butyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate 5 g, 17.18 mmol, 1.0 eq.
  • IBX 5.75 g, 20.61 mmol, 1.2 eq.
  • the reaction mixture was stirred at 60°C for 2 hours. After the reaction was completed, it was cooled to room temperature, diluted with H 2 O (500 mL), and extracted with EA (100 mL*3).
  • tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (1-5) (132 mg, 0.46 mmol, 0.96 eq.) and TEA (18 drops) were added to a solution of 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione (1-4) (180 mg, 0.48 mmol, 1.0 eq.) in DCM/MeOH (6 mL/4 mL).
  • NaBH(OAc) 3 270 mg, 1.30 mmol, 2.7 eq.
  • N,N-diethyl-4,5-difluoro-2-vinylbenzamide (3-3) (10.5 g, 44 mmol, 1.0 eq.), K 2 OsO4 ⁇ 2H 2 O (0.83 g, 1.7 mmol, 0.04 eq.) and NaIO4 (18.2 g, 110 mmol, 2.5 eq.) were added to 1,4-dioxane (100 mL) and H 2 O (40 mL) in sequence. After stirring at room temperature for 2 hours, the mixture was filtered and concentrated under reduced pressure. The residue was then added to 150 mL of EtOAc and 45 mL of H 2 O.
  • MsCl (3.1 g, 26.8 mmol, 1.8 eq.0) was added to a DCM (30 mL) solution of tert-butyl (R)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (3.0 g, 14.9 mmol, 1.0 eq.) and TEA (3.01 g, 29.8 mmol, 2.0 eq.). After stirring at room temperature for 4 hours, 30 mL of saturated brine was added to wash the mixture.
  • N-((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-4-(4-(hydroxymethyl)piperidin-1-yl)benzamide 9-4) (1.1 g, 2.4 mmol, 1.0 eq.) in DCM (120 mL) was added Dess-Martin reagent (1.2 g, 2.9 mmol, 1.2 eq.) at room temperature. After stirring at 25°C for 16 hours, the mixture was diluted with saturated aqueous Na2S2O3 solution (60 mL) and extracted with DCM (40 mL * 2).
  • N-((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (9-5) (108 mg, 0.24 mmol, 0.9 eq.) and TEA (6 drops) were added to 3-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione hydrochloride (9-9) (90 mg, 0.26 mmol, 1.0 eq.) in DCM/MeOH (10 mL/5 mL) at room temperature.
  • reaction mixture was purified by reverse phase chromatography (TFA buffer) to give tert-butyl 4-(1-(2,6-dioxopiperidin-3-yl)indolyl-5-yl)piperazine-1-carboxylate (10-3) (70 mg).
  • Triton-B 400 mg, 40%, 0.96 mmol, 0.7 eq. was added to a solution of 3-(1-(6-bromo-1-methyl-1H-indazol-3-yl)ureido)propanoate (11-4) (485 mg, 1.37 mmol, 1.0 eq.) in MeCN (5 mL) at room temperature. After stirring at 25°C for 16 h, EA (50 mL) and water (50 mL) were added. The organic layer was concentrated to give 1-(6-bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (11-5) (364 mg).
  • N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (103 mg, 0.21 mmol, 1.0 eq.) and 1-(1-methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (11-7) (70 mg, 0.21 mmol, 1.0 eq.) in DCM (5 ml) was added TEA (216 mg, 2.13 mmol, 10.0 eq.) and MgSO 4 (256 mg, 2.13 mmol, 10.0 eq.) at room temperature.
  • 3-Bromopiperidine-2,6-dione (183 mg, 0.96 mmol, 2.0 eq.) and NaHCO 3 (121 mg, 1.44 mmol, 3.0 eq.) were added to a solution of tert-butyl 4-(3-amino-5-chlorophenyl)piperazine- 1 -carboxylate (12-2) (150 mg, 0.48 mmol, 1.0 eq.) in DMF (2 mL) at room temperature.
  • N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (37 mg, 0.077 mmol, 1 eq.) in DCM (1 mL) at room temperature were added 3-((3-chloro-5-(piperazin-1-ylphenyl)amino)piperidine-2,6-dione (12-4) (25.0 mg, 0.077 mmol, 1.0 eq.), TEA (3 drops) and MgSO 4 (93 mg, 0.077 mmol, 1 eq.). 0.07 mmol, 10.0 eq.).
  • Triton-B (409 mg, 40%, 2.44 mmol, 2.5 eq.) was added to a solution of benzyl 4-(4-(1-(3-(3-methoxy-3-oxopropyl)ureido)-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-4) (420 mg, 0.97 mmol, 1.0 eq.) in MeCN (12 mL) at room temperature. After stirring at 25°C for 16 h, EA (50 mL) and water (50 mL) were added.
  • the mixture was stirred at 25°C for 2 hours, and then NaBH(OAc) was added . (120.1 mg, 0.5 mmol, 3.0 eq.).
  • the reaction mixture was stirred at 25°C for 16 hours, and DCM (20 mL) and water (20 mL) were added.
  • Boc 2 O (3.6 g, 16.4 mmol, 1.3 eq.) was added to a solution of 6-chloropyridazine-3-carboxylic acid (2 g, 12.6 mmol, 1.0 eq.) and DMAP (776 mg, 6.3 mmol, 0.5 eq.) in THF (50 mL).
  • H 2 O 400 mL was added and the mixture was extracted with EA (100 mL*2). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo.
  • tert-butyl 6-(4-formylpiperidin-1-yl)pyridazine-3-carboxylate (15-3) (80 mg).
  • N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-(hydroxymethyl)piperidin-1-yl)pyrazine-2-carboxamide (17-3) (100 mg, 0.2 mmol, 1.0 eq.) in DCM (3 mL) was added Dess-Martin reagent (104 mg, 0.26 mmol, 1.3 eq.) at room temperature. After stirring at 25°C for 3 hours, the mixture was diluted with saturated aqueous Na2S2O3 solution (25 mL ) and extracted with DCM (25 mL*3).
  • Pd(dppf)Cl 2 ⁇ CH 2 Cl 2 (15 mg, 0.02 mmol, 0.08 eq.) and KOAc (44 mg, 0.44 mmol, 2.0 eq.) were added to a solution of 6-bromophthalazin-1(2H)-one (50 mg, 0.22 mmol, 1.0 eq.) and B 2 Pin 2 ( 68 mg , 0.26 mmol, 1.2 eq.) in dioxane (2 ml) at room temperature. After stirring at 100° C. for 16 hours, the mixture was cooled to room temperature and concentrated.
  • Triton-B (492 mg, 40% in water, 1.18 mmol, 0.3 eq.) was added to a solution of methyl 3-(1-(1-methyl-7-nitro-1H-indazol-3-yl)ureido)propanoate (23-1-4) (1.26 g, 3.92 mmol, 1.0 eq.) in MeCN (13 mL). The reactant was stirred at 15 ° C for 72 hours. TLC showed that the reaction was complete. Water (100 mL) and EA (100 mL) were added.
  • Triton-B (246 mg, 0.59 mmol, 1.0 eq.) was added to a solution of benzyl 4-(5-(1-(3-(3-methoxy-3-oxopropyl)ureido)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (27-4) (300 mg, 0.59 mmol, 1.0 eq.) in MeCN (3 mL) at room temperature. After stirring for 16 hours, EA (30 mL) and water (30 mL) were added. The organic layer was concentrated.
  • Pd/C (10 mg, 5%) was added to a solution of 4-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid benzyl ester (27-5) (84 mg, 0.176 mmol, 1.0 eq.) in EA (1 mL) at room temperature.
  • Example 35 Compounds 67 and 67A were synthesized by referring to the preparation of compounds 58 and 58A.
  • the nuclear magnetic data (H-NMR) of the compounds of the present invention are as follows:
  • Test Example 1 Detection of the AR protein degradation level of the compounds of the present invention in human prostate cancer cells LNCaP and VCaP
  • %Degradation 100 - (OD value of compound well - OD value of positive reference well) / (OD value of negative reference well - OD value of positive reference well) value) ⁇ 100.
  • Test Example 2 Human prostate cancer cell VCaP, Western blotting to determine compound-induced AR degradation
  • the revived cells were cultured in T25 flasks with DMEM Medium + 10% FBS + 1% P/S. When the cell confluence reached 80-90%, the cells were passaged. Before the experiment, VCaP cells were inoculated into 96-well cell culture plates, 3 ⁇ 10 5 cells were inoculated into each well, and 100 ⁇ L of culture medium was added. The cells were cultured in a 37°C, 5% CO 2 incubator overnight.
  • Cell lysis buffer Add one tablet of protease inhibitor and one tablet of phosphatase inhibitor to 20 mL of cell lysis buffer and mix gently until completely dissolved.
  • Electrophoresis buffer Dilute 20 ⁇ electrophoresis buffer to 1 ⁇ with ultrapure water.
  • Transfer buffer Dilute 10 ⁇ transfer buffer to 1 ⁇ with ultrapure water and add 20% methanol.
  • TBST buffer dilute 10 ⁇ electrophoresis buffer to 1 ⁇ with ultrapure water.
  • Blocking solution Dissolve 5 g BSA in 1 ⁇ TBST to 100 mL and store at 4°C.
  • AR primary antibody working solution add Androgen Receptor (D6F11) to 24 mL of blocking solution Rabbit mAb #5153 antibody 24 ⁇ L;
  • ⁇ -actin primary antibody working solution add 2.4 ⁇ L of ⁇ -Actin (13E5) Rabbit mAb #4970 antibody to 24 mL of blocking solution;
  • the tube was centrifuged inverted at 300 rpm for 30 seconds. 45 ⁇ L of cell lysis solution was added to each well and the tube was placed on ice for 20 minutes. The cell lysis solution was aspirated into a new centrifuge tube, 15 ⁇ L of 4 ⁇ sample preparation working solution was added to each sample, and the tube was heated at 70°C for 10 minutes. The remaining protein sample was stored at -80°C.
  • the ratio of AR/ ⁇ -actin is used as the corrected AR content of the sample well, and the AR content of the sample well without compound is used as 100% AR content, that is, 0% AR Degradation.
  • the degradation rate is calculated by dividing the AR content of the sample well without compound by the AR content of the sample well.
  • the DC50 values were calculated using a 4-parameter model using Graphpad 8.0 software. The results are shown in Table 3.
  • mice Male ICR mice were fasted overnight. Six mice were weighed before administration, and the dosage was calculated based on the body weight. Three mice were administered intravenously or orally.
  • the compound solvent was 5% DMSO + 10% solutol + 85% saline.
  • the compound was administered orally by gavage at 3 mg/kg and by tail vein injection at 1 mg/kg.
  • IV after administration 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h, 24h.
  • PO 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 24h.
  • Blood was collected from the jugular vein, and about 0.03mL of each sample was collected. Heparin sodium was used for anticoagulation and placed on ice after collection.
  • Plasma samples were stored in a -80°C refrigerator before analysis. While analyzing the samples, the intra-day accuracy of the quality control samples was evaluated, and the accuracy of more than 66.7% of the quality control samples was required to be between 80-120%.
  • Phoenix WinNonlin7.0 was used to calculate the pharmacokinetic parameters based on the blood drug concentration data at different time points, providing parameters such as AUC0-t, AUC0- ⁇ , MRT0- ⁇ , Cmax, Tmax, and T1/2 and their mean and standard deviation. The experimental results are shown in Table 4.

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Abstract

Provided are a cyanoquinoline-targeting protein degradation molecule, a preparation method therefor and a use thereof. Particularly, the present invention relates to a compound represented by general formula (I), a preparation method therefor, a pharmaceutical composition comprising the compound, and a use of the compound as a protein degradation targeting chimera of an androgen receptor in preparation of medicaments for treating related diseases (including prostate cancer, breast cancer, and spinal and bulbar muscular atrophy) controlled by the androgen receptor, wherein the definitions of the substituents in general formula (I) are the same as those in the description.

Description

氰基喹啉类靶向蛋白降解分子、其制备方法和应用Cyanoquinoline targeted protein degradation molecules, preparation methods and applications thereof

本申请要求申请日为2023年7月12日的中国专利申请2023108479013和申请日为2024年4月29日的中国专利申请2024105278854的优先权。本申请引用上述中国专利申请的全文。This application claims the priority of Chinese Patent Application No. 2023108479013 filed on July 12, 2023 and Chinese Patent Application No. 2024105278854 filed on April 29, 2024. This application cites the full text of the above Chinese patent application.

技术领域Technical Field

本发明属于医药领域，具体而言，本发明涉及氰基喹啉类靶向蛋白降解分子、其制备方法和应用。The present invention belongs to the field of medicine, and in particular, relates to cyanoquinoline targeted protein degradation molecules, and preparation methods and applications thereof.

背景技术Background Art

前列腺癌是一种常见的男性恶性肿瘤，发病率仅次于肺癌，是导致全球成人男性死亡的第二大杀手。目前，雄激素去势治疗(ADT)是早期前列腺癌主要治疗方法，包括手术去势法和药物去势法。但ADT不能治愈前列腺癌，经过14-30个月的中位治疗时间后，几乎所有患者都会逐渐发展为去势抵抗性前列腺癌(CRPC)，患者对ADT不再敏感，中位生存期小于20个月。在前列腺癌组织中，雄激素受体(AR)的增殖或突变能使其对血清中较低含量的雄激素敏感，是前列腺癌恶化的主要原因。目前，治疗CRPC的雄激素受体抑制剂代表药物有阿比特龙、恩杂鲁胺等。然而，15-25％的患者对阿比特龙和恩杂鲁胺等第二代激素治疗没有反应，大多数产生反应性的患者最终会产生严重的耐药性，导致预后不良。Prostate cancer is a common male malignancy, second only to lung cancer in incidence, and is the second leading cause of death among adult men worldwide. At present, androgen deprivation therapy (ADT) is the main treatment for early prostate cancer, including surgical castration and medical castration. However, ADT cannot cure prostate cancer. After a median treatment time of 14-30 months, almost all patients will gradually develop castration-resistant prostate cancer (CRPC), and patients are no longer sensitive to ADT, with a median survival of less than 20 months. In prostate cancer tissue, the proliferation or mutation of androgen receptor (AR) can make it sensitive to lower levels of androgens in serum, which is the main cause of prostate cancer deterioration. At present, representative androgen receptor inhibitors for the treatment of CRPC include abiraterone, enzalutamide, etc. However, 15-25% of patients do not respond to second-generation hormone therapy such as abiraterone and enzalutamide, and most responsive patients will eventually develop severe drug resistance, leading to poor prognosis.

泛素-蛋白酶体系统(ubiquitin-proteasome system,UPS)是细胞中蛋白降解的主要途径，参与细胞中80％以上的蛋白质的降解。UPS由泛素(Ub)、泛素活化酶(E1)、泛素结合酶(E2)、泛素-蛋白质连接酶(E3)、26S蛋白酶体及其目标蛋白组成。泛素活化酶系统负责活化泛素，并将其结合到待降解的蛋白上，形成靶蛋白多聚泛素链，即泛素化。蛋白酶体系统可以识别已泛素化的蛋白并将其降解。蛋白水解靶向嵌合分子(protein proteolysis-targeting chimeras,PROTACs)是一种杂合双功能小分子化合物，包括一个能够与目标靶蛋白结合的小分子化合物，在其合适位置上连接基团，再与一个能够与E3泛素连接酶结合的小分子化合物连接。PROTACs通过将目标靶蛋白和细胞内的E3拉近，形成靶蛋白-PROTACs-E3三元聚合体，通过E3泛素连接酶给目标靶蛋白加上泛素化蛋白标签，利用泛素-蛋白酶体系统(ubiquitin-proteasome system，UPS)特异性地降解靶蛋白。The ubiquitin-proteasome system (UPS) is the main pathway for protein degradation in cells, and is involved in the degradation of more than 80% of proteins in cells. UPS consists of ubiquitin (Ub), ubiquitin activating enzyme (E1), ubiquitin conjugating enzyme (E2), ubiquitin-protein ligase (E3), 26S proteasome and its target proteins. The ubiquitin activating enzyme system is responsible for activating ubiquitin and binding it to the protein to be degraded to form a polyubiquitin chain of the target protein, i.e. ubiquitination. The proteasome system can recognize ubiquitinated proteins and degrade them. Protein proteolysis-targeting chimeras (PROTACs) are hybrid bifunctional small molecule compounds, including a small molecule compound that can bind to a target protein, a linker group at a suitable position, and a small molecule compound that can bind to an E3 ubiquitin ligase. PROTACs work by bringing the target protein and the E3 in the cell together to form a target protein-PROTACs-E3 ternary polymer, adding a ubiquitinated protein tag to the target protein through the E3 ubiquitin ligase, and using the ubiquitin-proteasome system (UPS) to specifically degrade the target protein.

利用PROTACs策略，制备出能够靶向识别/结合雄激素受体的蛋白降解靶向嵌合体,能够通过细胞内泛素-蛋白酶体降解系统而调控雄激素受体的水平,诱导雄激素受体降解,从而达到治疗前列腺癌等受雄激素受体调控的相关疾病的效果。Using the PROTACs strategy, we prepared a protein degradation targeted chimera that can target and recognize/bind to the androgen receptor. It can regulate the level of androgen receptor through the intracellular ubiquitin-proteasome degradation system and induce androgen receptor degradation, thereby achieving the effect of treating prostate cancer and other related diseases regulated by the androgen receptor.

所以,研发出一种能够与雄激素受体靶向结合、有效降解雄激素受体的双功能嵌合体分子，在治疗受雄激素受体调控的相关疾病上具有良好的应用前景。Therefore, the development of a bifunctional chimeric molecule that can target and bind to the androgen receptor and effectively degrade the androgen receptor has good application prospects in the treatment of related diseases regulated by the androgen receptor.

发明内容Summary of the invention

本发明提供一种通式(I)所示化合物、其立体异构体或其药学上可接受的盐：
The present invention provides a compound represented by general formula (I), its stereoisomers or pharmaceutically acceptable salts thereof:

其中：in:

M1选自N或CR1；M 1 is selected from N or CR 1 ;

M2选自N或CR2；M 2 is selected from N or CR 2 ;

M3选自N或CR3；M 3 is selected from N or CR 3 ;

M4选自N或CR4；M 4 is selected from N or CR 4 ;

M5选自N或CR5；M 5 is selected from N or CR 5 ;

R1、R2、R3、R4和R5各自独立的选自氢、氘、卤素、氰基、烷基、烷氧基、卤代烷基、卤代烷氧基、羟基烷基、羟基烷氧基、环烷基或杂环基；R 1 , R 2 , R 3 , R 4 and R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, hydroxyalkoxy, cycloalkyl or heterocyclyl;

环A、环B和环C各自独立的选自环烷基、杂环基、芳基或杂芳基；Ring A, Ring B and Ring C are each independently selected from cycloalkyl, heterocyclyl, aryl or heteroaryl;

L1选自键、-C(O)NH-、-NH-、-O-、-NHC(O)-、-C(O)-、-S(＝O)-、-S(＝O)2-、-CH2-或-(CH2)2-；L 1 is selected from a bond, -C(O)NH-, -NH-, -O-, -NHC(O)-, -C(O)-, -S(=O)-, -S(=O) 2 -, -CH 2 - or -(CH 2 ) 2 -;

L2为-Ak1-Cy1-Ak2-Cy2-Ak3-， L2 is -Ak1-Cy1-Ak2-Cy2-Ak3-,

Ak1、Ak2和Ak3各自独立的选自键、-(CH2)n1-O-(CH2)n2-、-(CH2)n3-、-(CH2)n4-S-(CH2)n5、-(CH2)n6-C(O)NH-(CH2)n7-、-(CH2)n8-NH-(CH2)n9、-(CH2)n10-NHC(O)-(CH2)n11、-(CH2)n12-C(O)-(CH2)n13、-(CH2)n14-S(＝O)-(CH2)n15-或-(CH2)n16-S(＝O)2-(CH2)n17-，所述的-(CH2)n1-O-(CH2)n2-、-(CH2)n3-、-(CH2)n4-S-(CH2)n5、-(CH2)n6-C(O)NH-(CH2)n7-、-(CH2)n8-NH-(CH2)n9、-(CH2)n10-NHC(O)-(CH2)n11、-(CH2)n12-C(O)-(CH2)n13、-(CH2)n14-S(＝O)-(CH2)n15-和-(CH2)n16-S(＝O)2-(CH2)n17-，任选的可以进一步被取代；Ak1, Ak2 and Ak3 are each independently selected from a bond, -(CH 2 ) n1 -O-(CH 2 ) n2 -, -(CH 2 ) n3 -, -(CH 2 ) n4 -S-(CH 2 ) n5 , -(CH 2 ) n6 -C(O)NH-(CH 2 ) n7 -, -(CH 2 ) n8 -NH-(CH 2 ) n9 , -(CH 2 ) n10 -NHC(O)-(CH 2 ) n11 , -(CH 2 ) n12 -C(O)-(CH 2 ) n13 , -(CH 2 ) n14 -S(＝O)-(CH 2 ) n15 - or -(CH 2 ) n16 -S(＝O) 2 -(CH 2 ) n17 -, wherein -(CH 2 ) n1 -O-(CH 2 ) n2 - 2 ) n2 -, -(CH 2 ) n3 -, -(CH 2 ) n4 -S-(CH 2 ) n5 , -(CH 2 ) n6 -C(O)NH-(CH 2 ) n7 -, -(CH 2 ) n8 -NH-(CH 2 ) n9 , -(CH 2 ) n10 -NHC(O)-(CH 2 ) n11 , -(CH 2 ) n12 -C(O)-(CH 2 ) n13 , -(CH 2 ) n14 -S(═O)-(CH 2 ) n15 - and -(CH 2 ) n16 -S(═O) 2 -(CH 2 ) n17 -, which may be further substituted;

Cy1和Cy2各自独立的选自键、亚环己基、亚哌啶基、亚哌嗪基、亚氮杂环丁烷基、亚氮杂环戊烷基、亚环丁烷基或亚环戊烷基，所述亚环己基、亚哌啶基、亚哌嗪基、亚氮杂环丁烷基、亚氮杂环戊烷基、亚环丁烷基和亚环戊烷基，任选的可以进一步被取代；或者，所述亚环己基、亚哌啶基、亚哌嗪基、亚氮杂环丁烷基、亚氮杂环戊烷基、亚环丁烷基和亚环戊烷基上的任意两个原子可以链接形成环烷基或杂环基；Cy1 and Cy2 are each independently selected from a bond, a cyclohexylene, a piperidinylene, a piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene or a cyclopentylene, and the cyclohexylene, piperidinylene, piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene and a cyclopentylene may optionally be further substituted; or, any two atoms on the cyclohexylene, piperidinylene, piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene and a cyclopentylene may be linked to form a cycloalkyl or heterocyclic group;

Ra、Rb和Rc各自独立的选自氢、氘、卤素、氧代基、氰基、羟基、烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基、杂环基、芳基或杂芳基，所述烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基、杂环基、芳基和杂芳基，任选的可以进一步被取代； Ra , Rb and Rc are each independently selected from hydrogen, deuterium, halogen, oxo, cyano, hydroxy, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, and the alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl may be further substituted;

或者，任意两个Rc链接形成环烷基、杂环基、芳基或杂芳基，所述环烷基、杂环基、芳基和杂芳基，任选的可以进一步被取代；Alternatively, any two R c are linked to form a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, and the cycloalkyl group, the heterocyclic group, the aryl group and the heteroaryl group may be further substituted;

或者，Ra与L1链接形成环烷基、杂环基或杂芳基，所述环烷基、杂环基和杂芳基，任选的可以进一步被取代；Alternatively, R a and L 1 are linked to form a cycloalkyl group, a heterocyclic group or a heteroaryl group, and the cycloalkyl group, the heterocyclic group and the heteroaryl group may be further substituted;

x、y和z各自独立的选自0、1、2或3；x, y and z are each independently selected from 0, 1, 2 or 3;

n1、n2、n3、n4、n5、n6、n7、n8、n9、n10、n11、n12、n13、n14、n15、n16和n17各自独立的 选自0、1、2或3。n1, n2, n3, n4, n5, n6, n7, n8, n9, n10, n11, n12, n13, n14, n15, n16 and n17 are independent Selected from 0, 1, 2 or 3.

在本发明的一个实施方案中，上文所述通式(I)进一步如通式(II)所示：
In one embodiment of the present invention, the general formula (I) described above is further represented by the general formula (II):

其中：Rd选自氢、卤素或C1-3烷基；p1和p2各自独立的选自0、1、2、3或4；L4选自键、-CH2-、-NH-、-O-、-S-或-C(O)-。wherein: R d is selected from hydrogen, halogen or C 1-3 alkyl; p1 and p2 are each independently selected from 0, 1, 2, 3 or 4; L 4 is selected from a bond, -CH 2 -, -NH-, -O-, -S- or -C(O)-.

在发明的一个实施方案中，中的M2、M3、M4和M5不同时为N。In one embodiment of the invention, M 2 , M 3 , M 4 and M 5 are not N at the same time.

在本发明优选的实施方式中，上文所述环A选自苯基、5-6元单环杂芳基或9-10元双环杂环基；In a preferred embodiment of the present invention, the ring A mentioned above is selected from phenyl, 5-6 membered monocyclic heteroaryl or 9-10 membered bicyclic heterocyclic group;

优选的，环A选自苯基、吡唑基、吡咯基、吡啶基、嘧啶基、苯并哒嗪酮基、苯并吡唑基或苯并吡咯烷基；Preferably, ring A is selected from phenyl, pyrazolyl, pyrrolyl, pyridyl, pyrimidinyl, benzopyridazinone, benzopyrazolyl or benzopyrrolidinyl;

或者，环B选自环己烷基或环丁烷基；Alternatively, Ring B is selected from cyclohexyl or cyclobutane;

或者，环C选自苯基、5-6元单环杂芳基、9-10元双环杂环基、9-10元双环杂芳基；Alternatively, ring C is selected from phenyl, 5-6 membered monocyclic heteroaryl, 9-10 membered bicyclic heterocyclyl, 9-10 membered bicyclic heteroaryl;

优选的，环C选自苯并吡啶基、苯基、苯并噻唑基、苯并吡嗪基、吡啶并吡唑基、吡啶并三唑基、吡啶并苯基、苯并噻二唑或苯并哒嗪基。Preferably, ring C is selected from benzopyridyl, phenyl, benzothiazolyl, benzopyrazinyl, pyridopyrazolyl, pyridotriazolyl, pyridophenyl, benzothiadiazole or benzopyridazinyl.

在本发明优选的实施方式中，上文所述化合物进一步如通式(II-A)所示：
In a preferred embodiment of the present invention, the compound described above is further represented by general formula (II-A):

其中：in:

选自且， Selected from and,

当选自时，环A不为 when Selected from When ring A is not

在本发明优选的实施方式中，上文所述环A选自苯基、吡啶基、 In a preferred embodiment of the present invention, the ring A is selected from phenyl, pyridyl,

在本发明优选的实施方式中，上文所述通式(I)或通式(II)中的选自 In a preferred embodiment of the present invention, in the above-mentioned general formula (I) or general formula (II) Selected from

在本发明优选的实施方式中，上文所述Ra、Rb和Rc各自独立的选自氢、氘、卤素、氰基、羟基、氧代基、C1-3烷基、C1-3烷氧基、羟基C1-3烷基、羟基C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C3- 4环烷基或3-4元杂环基。In a preferred embodiment of the present invention, Ra , Rb and Rc described above are each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, oxo, C1-3 alkyl, C1-3 alkoxy, hydroxyC1-3 alkyl, hydroxyC1-3 alkoxy, C1-3 haloalkyl, C1-3 haloalkoxy, C3-4 cycloalkyl or 3-4 membered heterocyclic group.

在本发明优选的实施方式中，上文所述R1、R2、R3、R4和R5各自独立的选自氢、氘、卤素、氰基、C1-3烷基、C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、羟基C1-3烷基、羟基C1-3烷氧基、C3-4环烷基或3-4元杂环基。In a preferred embodiment of the present invention, R 1 , R 2 , R 3 , R 4 and R 5 mentioned above are each independently selected from hydrogen, deuterium, halogen, cyano, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, hydroxy C 1-3 alkyl, hydroxy C 1-3 alkoxy, C 3-4 cycloalkyl or 3-4 membered heterocyclic group.

在本发明优选的实施方式中，上文所述L1选自键、-C(O)NH-、-NH-、-O-、-NHC(O)-、-C(O)-、-CH2-或-(CH2)2-；In a preferred embodiment of the present invention, L 1 mentioned above is selected from a bond, -C(O)NH-, -NH-, -O-, -NHC(O)-, -C(O)-, -CH 2 - or -(CH 2 ) 2 -;

L2选自 L 2 is selected from

在本发明优选的实施方式中，上文所述化合物进一步如通式(II-B)所示：
In a preferred embodiment of the present invention, the compound described above is further represented by general formula (II-B):

其中：in:

L3和L4各自独立的选自键、-CH2-、-NH-、-O-、-S-或-C(O)-；L 3 and L 4 are each independently selected from a bond, -CH 2 -, -NH-, -O-, -S- or -C(O)-;

M6和M7各自独立的选自-CR6R7-或-C(O)-；M 6 and M 7 are each independently selected from -CR 6 R 7 - or -C(O)-;

M8选自N或CR8；M 8 is selected from N or CR 8 ;

R6、R7和R8各自独立的选自氢、氘、氟、氯、羟基、氰基、C1-3烷基、羟基C1-3烷基、C1-3烷氧基、羟基C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C3-4环烷基或3-4元杂环基；R 6 , R 7 and R 8 are each independently selected from hydrogen, deuterium, fluorine, chlorine, hydroxyl, cyano, C 1-3 alkyl, hydroxy C 1-3 alkyl, C 1-3 alkoxy, hydroxy C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, C 3-4 cycloalkyl or 3-4 membered heterocyclyl;

j、p1和p2各自独立的选自0、1、2、3或4。j, p1 and p2 are each independently selected from 0, 1, 2, 3 or 4.

在本发明的一个具体实施方案中，本发明上文所述通式(II)化合物进一步如通式(II-C)所示：
In a specific embodiment of the present invention, the compound of the general formula (II) described above is further represented by the general formula (II-C):

其中：M2选自N或CR2；M3选自N或CR3；M4选自N或CR4；M5选自N或CR5；环B选自环已烷基或环丁烷基；环C选自苯基、5-6元单环杂芳基或10元双环杂芳基；优选地，环C选自苯基、吡啶基或苯并吡啶基；Ra选自氢、卤素或C1-3烷基；Rd选自氢、卤素或C1-3烷基；Rb各自独立地选自氢、C1-3烷基、C1-3烷氧基或C1-3羟基烷基；Rc各自独立地选自氢、氰基、卤素、羟基、C1-3烷基、C1- 3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C1-3羟基烷基或C1-3羟基烷氧基；R2、R3、R4和R5各自独立地选自氢、卤素、氰基、C1-3烷基、C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C1-3羟基烷基或C1-3羟基烷氧基；y和z各自独立的选自0、1、2或3。wherein: M 2 is selected from N or CR 2 ; M 3 is selected from N or CR 3 ; M 4 is selected from N or CR 4 ; M 5 is selected from N or CR 5 ; Ring B is selected from cyclohexyl or cyclobutane; Ring C is selected from phenyl, 5-6-membered monocyclic heteroaryl or 10-membered bicyclic heteroaryl; preferably, Ring C is selected from phenyl, pyridyl or benzopyridyl; Ra is selected from hydrogen, halogen or C 1-3 alkyl; R d is selected from hydrogen, halogen or C 1-3 alkyl; R b is each independently selected from hydrogen, C 1-3 alkyl, C 1-3 alkoxy or C 1-3 hydroxyalkyl; R c is each independently selected from hydrogen, cyano, halogen, hydroxyl, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, C 1-3 hydroxyalkyl or C 1-3 hydroxyalkoxy; R 2 , R 3 , R 4 and R 5 are each independently selected from hydrogen, halogen, cyano, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, C 1-3 hydroxyalkyl or C 1-3 hydroxyalkoxy; y and z are each independently selected from 0, 1, 2 or 3.

在本发明的一个具体实施方案中，本发明上文所述Ra选自氢、氟、氯、甲基或乙基。In a specific embodiment of the present invention, Ra described above in the present invention is selected from hydrogen, fluorine, chlorine, methyl or ethyl.

在本发明的一个具体实施方案中，本发明上文所述Rb各自独立地选自氢、甲基、乙基、甲氧基、乙氧基、羟基甲基或羟基乙基。In a specific embodiment of the present invention, the R b mentioned above in the present invention is independently selected from hydrogen, methyl, ethyl, methoxy, ethoxy, hydroxymethyl or hydroxyethyl.

在本发明的一个具体实施方案中，本发明上文所述Rc各自独立地选自氢、氰基、氟、氯、羟基、甲基、乙基、甲氧基、乙氧基、三氟甲基、三氟乙基、羟基甲基或羟基乙基。In a specific embodiment of the present invention, R c mentioned above in the present invention is independently selected from hydrogen, cyano, fluorine, chlorine, hydroxyl, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, hydroxymethyl or hydroxyethyl.

在本发明的一个具体实施方案中，本发明上文所述Rd选自氢、氟、氯、甲基或乙基。In one embodiment of the present invention, R d as described above in the present invention is selected from hydrogen, fluorine, chlorine, methyl or ethyl.

在本发明的一个具体实施方案中，本发明上文所述R2、R3、R4和R5各自独立地选自氢、氟、氯、氰基、甲基、乙基、甲氧基、乙氧基、三氟甲基、三氟乙基、羟基甲基或羟基乙基。 In a specific embodiment of the present invention, R 2 , R 3 , R 4 and R 5 described above are each independently selected from hydrogen, fluorine, chlorine, cyano, methyl, ethyl, methoxy, ethoxy, trifluoromethyl, trifluoroethyl, hydroxymethyl or hydroxyethyl.

在本发明的一个优选实施方案中，本发明上文所述环选自 In a preferred embodiment of the present invention, the above-mentioned ring Selected from

在本发明的一个优选实施方案中，本发明上文所述为 In a preferred embodiment of the present invention, the present invention described above for

在本发明优选的实施方式中，上文所述化合物选自如下表5化合物：In a preferred embodiment of the present invention, the above-mentioned compound is selected from the following compounds in Table 5:

表5









Table 5









本发明提供一种药物组合物，包括治疗有效量的上文所述的化合物、其立体异构体或其药学上可接受的盐，以及一种或多种药学上可接受的载体。The present invention provides a pharmaceutical composition comprising a therapeutically effective amount of the above-mentioned compound, its stereoisomer or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers.

本发明还涉及一种优选方案，涉及各通式化合物、其立体异构体或其药学上可接受的盐，或上文所述的组合物，在制备雄激素受体调控的相关疾病的药物中的应用。The present invention also relates to a preferred embodiment, involving the use of each compound of the general formula, its stereoisomers or pharmaceutically acceptable salts, or the composition described above in the preparation of drugs for androgen receptor-regulated related diseases.

进一步地，上文所述疾病选自前列腺癌、乳腺癌、肯尼迪氏病。Furthermore, the above-mentioned disease is selected from prostate cancer, breast cancer, and Kennedy's disease.

本发明还提供了一种治疗受雄激素受体调控的相关疾病的药物，所述药物是以上述的化合物、或其同位素化合物、或其光学异构体、或其互变异构体、或其药学上可接受的盐、或其前药、或其溶剂合物为活性成分，加上药学上可接受的辅料制得的制剂。The present invention also provides a drug for treating diseases regulated by androgen receptors, wherein the drug is a preparation prepared with the above-mentioned compound, or its isotope compound, or its optical isomer, or its tautomer, or its pharmaceutically acceptable salt, or its prodrug, or its solvate as an active ingredient and a pharmaceutically acceptable excipient.

本发明还提供一种式(A-1)所示化合物、其立体异构体或其药学上可接受的盐：
The present invention also provides a compound represented by formula (A-1), a stereoisomer thereof or a pharmaceutically acceptable salt thereof:

其中：R10选自氢或氨基保护基团；环A、M1、L1、L4、Ra、p1、p2和x如本发明前文所定义。Wherein: R 10 is selected from hydrogen or an amino protecting group; Ring A, M 1 , L 1 , L 4 , Ra , p1, p2 and x are as defined above in the present invention.

在本发明中，氨基保护基团优选为SEM、叔丁氧羰基、-CH2-叔丁氧羰基、-CH2COOH、-COCH3、乙氧基羰基、苄基、对甲氧基苄基、苄氧羰基、芴甲氧羰基或烯丙氧羰基。In the present invention, the amino protecting group is preferably SEM, tert-butyloxycarbonyl, -CH2 -tert-butyloxycarbonyl, -CH2COOH , -COCH3 , ethoxycarbonyl, benzyl, p-methoxybenzyl, benzyloxycarbonyl, fluorenylmethoxycarbonyl or allyloxycarbonyl.

本发明还涉及式(A-1-1)的结构单元作为蛋白降解抑制剂结构单元中的应用：
The present invention also relates to the use of the structural unit of formula (A-1-1) as a structural unit of a protein degradation inhibitor:

其中：环A、M1、L1、L4、Ra、p1、p2和x如本发明前文所定义。 wherein: ring A, M 1 , L 1 , L 4 , Ra , p1, p2 and x are as defined above in the present invention.

在本发明优选的实施方式中，所述式(A-1)化合物进一步如式(A-1-a)所示：
In a preferred embodiment of the present invention, the compound of formula (A-1) is further represented by formula (A-1-a):

在本发明优选的实施方式中，所述式(A-1-1)进一步如式(A-1-1-a)所示：
In a preferred embodiment of the present invention, the formula (A-1-1) is further shown as formula (A-1-1-a):

通过实验证明，本发明提供的式(I)所示的化合物能够靶向降解前列腺癌细胞中的雄激素受体，并且抑制前列腺癌细胞的增殖，同时还显示了良好的代谢稳定性和药代动力学性质。本发明化合物在制备雄激素受体的蛋白降解靶向嵌合体，以及制备治疗受雄激素受体调控的相关疾病(包括前列腺癌、乳腺癌、肯尼迪氏病)的药物中具有良好的应用前景。Experiments have shown that the compound of formula (I) provided by the present invention can target androgen receptors in prostate cancer cells and inhibit the proliferation of prostate cancer cells, while also showing good metabolic stability and pharmacokinetic properties. The compound of the present invention has good application prospects in the preparation of protein degradation targeted chimeras of androgen receptors, and in the preparation of drugs for treating related diseases regulated by androgen receptors (including prostate cancer, breast cancer, and Kennedy's disease).

发明的详细说明Detailed description of the invention

除非有相反陈述，在说明书和权利要求书中使用的术语具有下述含义。Unless stated otherwise, the terms used in the specification and claims have the following meanings.

术语“烷基”指饱和脂肪族烃基团，其为包含1至20个碳原子的直链或支链基团，优选含有1至8个碳原子的烷基，更优选1至6个碳原子的烷基，最更优选1至3个碳原子的烷基。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、正己基等，及其各种支链异构体等。本发明优选甲基、乙基、异丙基、叔丁基、卤代烷基、氘代烷基、烷氧基取代的烷基和羟基取代的烷基。The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably an alkyl group containing 1 to 6 carbon atoms, and most preferably an alkyl group containing 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, etc., and various branched isomers thereof, etc. Methyl, ethyl, isopropyl, tert-butyl, haloalkyl, deuterated alkyl, alkoxy-substituted alkyl, and hydroxy-substituted alkyl are preferred in the present invention.

术语“亚烷基”是指烷基的一个氢原子进一步被取代，例如：“亚甲基”指-CH2-、“亚乙基”指-(CH2)2-、“亚丙基”指-(CH2)3-、“亚丁基”指-(CH2)4-等。The term "alkylene" refers to an alkyl group in which one hydrogen atom is further substituted. For example, "methylene" refers to -CH2- , "ethylene" refers to -( CH2 ) 2- , "propylene" refers to -( CH2 ) 3- , "butylene" refers to -( CH2 ) 4- , and the like.

术语“烯基”指由至少两个碳原子和至少一个碳-碳双键组成的如上定义的烷基，例如乙烯基、1-丙烯基、2-丙烯基、1-、2-或3-丁烯基等。烯基可以是取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基。The term "alkenyl" refers to an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, such as vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3-butenyl, etc. Alkenyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio.

术语“亚烯基”是指烯基的一个氢原子进一步被取代，例如：“亚乙烯基”指-(CH)2-。The term "alkenylene" refers to an alkenyl group in which one of its hydrogen atoms is further substituted, for example: "ethenylene" refers to -(CH) 2 -.

术语“环烷基”指饱和或部分不饱和单环或多环环状烃取代基，环烷基环包含3至20个碳原子，优选包含3至12个碳原子，更优选包含3至6个碳原子，进一步优选包含3至4各碳原子。单环环烷基的非限制性实例包括环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基、环庚三烯基、环辛基等；多环环烷基包括螺环、稠环和桥环的环烷基，非限制性实例包括 所述环烷基环可以稠合于芳基、杂芳基或杂环烷基环上，其中与母体结构连接在一起的环为环烷基，非限制性实例包括茚满基、四氢萘基、苯并环庚烷基等。环烷基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氧代基、羧基或羧酸酯基。The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, more preferably 3 to 6 carbon atoms, and further preferably 3 to 4 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl, cyclooctyl, etc.; polycyclic cycloalkyls include spirocyclic, fused and bridged cycloalkyls, non-limiting examples of which include The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring, wherein the ring attached to the parent structure is a cycloalkyl, non-limiting examples of which include indanyl, tetrahydronaphthyl, benzocycloheptanyl, etc. The cycloalkyl may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

术语“亚环烷基”指环烷基的一个氢原子进一步被取代，非限制性实例包括：亚环己基、 亚环烷基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氧代基、羧基或羧酸酯基。The term "cycloalkylene" refers to a cycloalkyl group in which one hydrogen atom is further substituted. Non-limiting examples include: cyclohexylene, The cycloalkylene group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

术语“杂环基”指饱和或部分不饱和单环或多环环状烃取代基，其包含3至20个环原子，其中一个或多个环原子为选自氮、氧、C(O)、S(O)(＝NH)或S(O)m(其中m是整数0至2)的杂原子，但不包括-O-O-、-O-S-或-S-S-的环部分，其余环原子为碳。优选包含3至12个环原子，其中1～4个是杂原子；更优选包含3至12个环原子；最优选包含3至6个环原子，进一步优选3至4个环原子。单环杂环基的非限制性实例包括氧杂环丁烷基、氮杂环丁烷基、四氢吡喃基基、氮杂环庚烷基、吡咯烷基、咪唑烷基、四氢呋喃基、四氢噻吩基、二氢咪唑基、二氢呋喃基、二氢吡唑基、二氢吡咯基、哌啶基、哌嗪基、吗啉基、硫代吗啉基、高哌嗪基、吡喃基等，优选氧杂环丁烷、硫杂环丁烷、氮杂环丁烷、四氢呋喃基、四氢吡喃基、1-氨亚基-1-氧代噻喃、氮杂环庚烷基、哌啶基和哌嗪基。多环杂环基包括螺环、稠环和桥环的杂环基，非限制性实例包括杂环基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、氨基、环烷基、杂环烷基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氧代基、羧基或羧酸酯基。The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 20 ring atoms, one or more of which is a heteroatom selected from nitrogen, oxygen, C(O), S(O)(=NH) or S(O) m (wherein m is an integer from 0 to 2), but excluding the ring part of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon. Preferably, it contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably, it contains 3 to 12 ring atoms; most preferably, it contains 3 to 6 ring atoms, and further preferably, it contains 3 to 4 ring atoms. Non-limiting examples of monocyclic heterocyclic groups include oxetane, azetane, tetrahydropyranyl, azepanyl, pyrrolidinyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuranyl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, pyranyl, etc., preferably oxetane, thietane, azetane, tetrahydrofuranyl, tetrahydropyranyl, 1-aminoylidene-1-oxothiopyran, azepanyl, piperidinyl and piperazinyl. Polycyclic heterocyclic groups include spirocyclic, condensed ring and bridged ring heterocyclic groups, non-limiting examples include The heterocyclic group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

术语“亚杂环基”是指杂环基的一个氢原子进一步被取代，其非限制性实例包括：亚哌啶基、亚哌嗪基、亚吡咯并吡咯基、亚二氮杂螺[5.5]十一烷基、亚氮杂螺[5.5]十一烷基、亚苯并哌啶基、亚氮杂环丁烷基、亚二氮杂环丁烷、亚吡咯烷基、亚氮杂螺环[3.5]壬烷基、亚二氮螺环[3.5]壬烷基、亚氮杂双环[3.1.1]庚烷基、亚氮杂螺环[2.5]辛烷基、亚杂环基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、氨基、环烷基、杂环烷基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氧代基、羧基或羧酸酯基。The term "heterocyclylene" refers to a heterocyclyl group in which one hydrogen atom is further substituted, and non-limiting examples thereof include: piperidinylene, piperazinylene, pyrrolopyrrolylene, diazaspiro[5.5]undecylene, azaspiro[5.5]undecylene, benzopiperidinylene, azetidinylene, diazetidinylene, pyrrolidinylene, azaspiro[3.5]nonanylene, diazaspiro[3.5]nonanylene, azabicyclo[3.1.1]heptanylene, azaspiro[2.5]octanylene, The heterocyclylene group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate.

术语“芳基”指具有共轭的π电子体系的6至14元全碳单环或稠合多环(也就是共享毗邻碳原子对的环)基团，优选为6至10元，更优选苯基。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。芳基可以是取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、羧基或羧酸酯基。The term "aryl" refers to a 6- to 14-membered all-carbon monocyclic or fused polycyclic (i.e., rings that share adjacent pairs of carbon atoms) group having a conjugated π electron system, preferably 6- to 10-membered, more preferably phenyl. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, wherein The ring connected to the parent structure is an aryl ring. The aryl group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

术语“亚芳基”是指芳基的一个氢原子进一步被取代，其非限制性实例包括：亚芳基可以是取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、羧基或羧酸酯基。The term "arylene" refers to an aryl group in which one hydrogen atom is further substituted, and non-limiting examples thereof include: The arylene group may be substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from alkyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

术语“杂芳基”指包含1至4个杂原子、5至14个环原子的杂芳族体系，其中杂原子选自氧、硫和氮。杂芳基优选为5-8元单杂芳基或8-14原双杂芳基，更优选为5元单杂芳基、6元单杂芳基或9元双杂芳基，例如咪唑基、呋喃基、噻吩基、噻唑基、吡唑基、吡咯基、三唑基、四唑基、吡啶基、嘧啶基、吡嗪基、哒嗪基、哌嗪基、吡啶并咪唑基、嘧啶并咪唑基等，优选吡啶并咪唑基、嘧啶并咪唑基。杂芳基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、氨基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、羧基或羧酸酯基。The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably a 5-8-membered monoheteroaryl or an 8-14-membered diheteroaryl, more preferably a 5-membered monoheteroaryl, a 6-membered monoheteroaryl or a 9-membered diheteroaryl, such as imidazolyl, furanyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, triazolyl, tetrazolyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, piperazinyl, pyridoimidazolyl, pyrimidoimidazolyl, etc., preferably pyridoimidazolyl and pyrimidoimidazolyl. The heteroaryl group may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxy, nitro, cyano, amino, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate.

术语“亚杂芳基”是指杂芳基的一个氢原子进一步被取代，其非限制性实例包括：亚吡啶基、亚嘧啶基、亚吡唑基、亚哒嗪基、亚吡嗪基、 The term "heteroarylene" refers to a heteroaryl group in which one hydrogen atom is further substituted, and non-limiting examples thereof include pyridylene, pyrimidylene, pyrazolylene, pyridazinylene, pyrazinylene,

术语“烷氧基”指-O-(烷基)和-O-(非取代的环烷基)，其中烷基的定义如上所述。烷氧基的非限制性实例包括：甲氧基、乙氧基、丙氧基、丁氧基、环丙氧基、环丁氧基、环戊氧基、环己氧基。烷氧基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、羧基或羧酸酯基。非限制性的实例包括三氟甲氧基、三氟乙氧基、二氟甲氧基。The term "alkoxy" refers to-O-(alkyl) and-O-(non-substituted cycloalkyl), wherein the definition of alkyl is as described above. The non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropyloxy, cyclobutyloxy, cyclopentyloxy, cyclohexyloxy. Alkoxy can be optionally substituted or non-substituted, and when substituted, substituents are preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, sulfydryl, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkyloxy, heterocycloalkyloxy, cycloalkylthio, heterocycloalkylthio, carboxyl or carboxylate. Non-limiting examples include trifluoromethoxy, trifluoroethoxy, difluoromethoxy.

“卤代烷基”指被一个或多个卤素取代的烷基，其中烷基如上所定义。非限制性实例包括：三氟甲基、二氟甲基。"Haloalkyl" refers to an alkyl group substituted with one or more halogens, wherein alkyl is as defined above. Non-limiting examples include: trifluoromethyl, difluoromethyl.

“卤代烷氧基”指被一个或多个卤素取代的烷氧基，其中烷氧基如上所定义。"Haloalkoxy" refers to an alkoxy group substituted with one or more halogens, wherein alkoxy is as defined above.

“羟基烷基”指被羟基取代的烷基，其中烷基如上所定义。非限制性实例包括：-C(CH3)2(OH)。"Hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above. Non-limiting examples include: -C(CH 3 ) 2 (OH).

“羟基烷氧基”指羟基取代的烷氧基，其中烷氧基如上所定义。"Hydroxyalkoxy" means an alkoxy group substituted with a hydroxy group wherein alkoxy is as defined above.

“X选自A、B、或C”、“X选自A、B和C”、“X为A、B或C”、“X为A、B和C”等不同用语均表达了相同的意义，即表示X可以是A、B、C中的任意一种或几种。Different expressions such as “X is selected from A, B, or C”, “X is selected from A, B and C”, “X is A, B or C”, “X is A, B and C” all express the same meaning, that is, X can be any one or more of A, B, C.

本发明中表示键或不存在；In the present invention Indicates that a key may not exist;

对于本发明化合物中的手性碳，除特别说明外，为R构型或S构型。Unless otherwise specified, the chiral carbon in the compounds of the present invention is in R configuration or S configuration.

本发明所述的氢均可被其同位素氘所取代，本发明涉及的实施例化合物中的任一氢也均可被氘原子取代。 The hydrogen atoms described in the present invention may be replaced by their isotope deuterium atoms, and any hydrogen atoms in the example compounds of the present invention may also be replaced by deuterium atoms.

“任选”或“任选地”意味着随后所描述的事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生的场合。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，该说明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。"Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "a heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes instances where the heterocyclic group is substituted with an alkyl group and instances where the heterocyclic group is not substituted with an alkyl group.

“取代的”指基团中的一个或多个氢原子，优选为最多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。"Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in the group are replaced independently of each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and the skilled person can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, amino or hydroxy groups with free hydrogens may be unstable when combined with carbon atoms with unsaturated (e.g. olefinic) bonds.

“任选的可以进一步被取代”中的取代基各自独立的选自氘、卤素、氰基、羟基、硝基、氨基、氧代基、烷基、烷氧基、卤代烷基、卤代烷氧基、羟基烷基、环烷基、杂环基、芳基或杂芳基；其中，卤素、氰基、羟基、硝基、氨基、氧代基、烷基、烷氧基、卤代烷基、卤代烷氧基、羟基烷基、环烷基、杂环基、芳基和杂芳基如上文所定义。The substituents in "optionally may be further substituted" are each independently selected from deuterium, halogen, cyano, hydroxy, nitro, amino, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl or heteroaryl; wherein, halogen, cyano, hydroxy, nitro, amino, oxo, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are as defined above.

“药物组合物”表示含有一种或多种本文所述化合物或其生理学上/可药用的盐或前体药物与其他化学组分的混合物，以及其他组分例如生理学/可药用的载体和赋形剂。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。"Pharmaceutical composition" means a mixture containing one or more compounds described herein or their physiologically/pharmaceutically acceptable salts or prodrugs and other chemical components, as well as other components such as physiologically/pharmaceutically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitate the absorption of the active ingredient, and thus exert biological activity.

“可药用盐”是指本发明化合物的盐，这类盐用于哺乳动物体内时具有安全性和有效性，且具有应有的生物活性。"Pharmaceutically acceptable salts" refer to salts of the compounds of the present invention, which are safe and effective when used in mammals and have the desired biological activity.

式(II)化合物通用合成方法：
General synthesis method of compound of formula (II):

在路线1中，式III的化合物中PG代表保护基，如叔丁基、甲基。如路线1所示，式III的化合物在碱性条件下(如用N,N-二异丙基乙胺做碱)，在合适的溶剂中如N,N-二甲基甲酰胺与式IV-A或IV-B的化合物发生芳香亲核取代反应，或利用钯催化剂如[1,1'-双(二苯基膦基)二茂铁]二氯化钯在合适的溶剂中如甲苯，和使用适当的碱如碳酸铯进行Buchwald-Hartwig偶联反应而得到式V-A或V-B的化合物。In route 1, PG in the compound of formula III represents a protecting group, such as tert-butyl or methyl. As shown in route 1, the compound of formula III undergoes an aromatic nucleophilic substitution reaction with a compound of formula IV-A or IV-B under alkaline conditions (such as using N,N-diisopropylethylamine as a base) in a suitable solvent such as N,N-dimethylformamide, or undergoes a Buchwald-Hartwig coupling reaction using a palladium catalyst such as [1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride in a suitable solvent such as toluene and using an appropriate base such as cesium carbonate to obtain a compound of formula V-A or V-B.

式V-A或V-B的化合物的脱保护产物式VI-A或VI-B的化合物利用常规的文献报道方法如用N,N-二异丙基乙胺做碱，2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU)或其他缩合试剂做活化试剂，与式VII的化合物反应得酰胺式VIII-A或VIII-B的化合物。 The deprotected product of the compound of formula VA or VB, the compound of formula VI-A or VI-B, is reacted with the compound of formula VII using conventional methods reported in the literature, such as using N,N-diisopropylethylamine as a base and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) or other condensation reagents as an activating reagent to obtain an amide compound of formula VIII-A or VIII-B.

式VIII-A的化合物经氧化(如用Dess-Martin试剂做氧化剂)得式IX的化合物；式VIII-B的化合物在酸性条件下(如盐酸)水解缩醛得式IX的化合物得式IX的化合物。式IX的化合物与式X的化合物在合适的还原剂下(如醋酸硼氢化钠)发生还原胺化反应得最终产物式II的化合物。The compound of formula VIII-A is oxidized (e.g., using Dess-Martin reagent as an oxidant) to obtain a compound of formula IX; the compound of formula VIII-B is hydrolyzed under acidic conditions (e.g., hydrochloric acid) to obtain a compound of formula IX. The compound of formula IX is subjected to a reductive amination reaction with a compound of formula X in the presence of a suitable reducing agent (e.g., sodium acetate borohydride) to obtain a final product, a compound of formula II.

或者利用常规的文献报道方法如用三乙胺做碱，用甲基磺酰氯将式VIII-A的化合物转化成式XI的磺酸酯化合物。式XI的化合物在碱性条件下(如用碳酸钾做碱)，在合适的溶剂中如N,N-二甲基甲酰胺与式X的化合物发生亲核取代反应得最终产物式II的化合物。
Alternatively, the compound of formula VIII-A can be converted into a sulfonate compound of formula XI by conventional methods reported in literature, such as using triethylamine as a base and methylsulfonyl chloride. The compound of formula XI undergoes a nucleophilic substitution reaction with the compound of formula X in a suitable solvent such as N,N-dimethylformamide under alkaline conditions (such as using potassium carbonate as a base) to obtain the final product, the compound of formula II.

另一条路线是将式V-A的化合物经氧化(如用Dess-Martin试剂做氧化剂)得式XII的化合物；式V-B的化合物在酸性条件下(如盐酸)水解缩醛得式XII的化合物。式XII的化合物与式X的化合物在合适的还原剂下(如醋酸硼氢化钠)发生还原胺化反应得式XIV的化合物。或利用常规的文献报道方法如用三乙胺做碱，用甲基磺酰氯将式V-A的化合物转化成式XIII的磺酸酯化合物。式XIII 的化合物在碱性条件下(如用碳酸钾做碱)，在合适的溶剂中如N,N-二甲基甲酰胺与式X的化合物发生亲核取代反应得式XIV的化合物。Another route is to oxidize the compound of formula VA (such as using Dess-Martin reagent as an oxidant) to obtain a compound of formula XII; the compound of formula VB is hydrolyzed under acidic conditions (such as hydrochloric acid) to obtain a compound of formula XII. The compound of formula XII and the compound of formula X are subjected to a reductive amination reaction under a suitable reducing agent (such as sodium acetate borohydride) to obtain a compound of formula XIV. Or, the compound of formula VA is converted into a sulfonate compound of formula XIII using conventional methods reported in the literature, such as using triethylamine as a base and methylsulfonyl chloride. Formula XIII The compound of formula X undergoes nucleophilic substitution reaction with the compound of formula X in a suitable solvent such as N,N-dimethylformamide under alkaline conditions (such as using potassium carbonate as a base) to obtain a compound of formula XIV.

式XIV的化合物的脱保护产物式XV的化合物利用常规的文献报道方法如用N,N-二异丙基乙胺做碱，2-(7-氮杂苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯(HATU)或其他缩合试剂，与式VII的化合物反应得最终产物式II的化合物。The deprotected product of the compound of formula XIV, the compound of formula XV, is reacted with the compound of formula VII using conventional methods reported in the literature, such as using N,N-diisopropylethylamine as a base, 2-(7-azabenzotriazole)-N,N,N',N'-tetramethyluronium hexafluorophosphate (HATU) or other condensation reagents to obtain the final product, the compound of formula II.

具体实施方式DETAILED DESCRIPTION

以下结合实施例进一步描述本发明，但这些实施例并非限制着本发明的范围。The present invention is further described below with reference to examples, but these examples are not intended to limit the scope of the present invention.

本发明实施例中的起始原料和实验所用试剂是已知的并且可以在市场上买到，或者可以采用或按照本领域已知的方法来合成。The starting materials and reagents used in the examples of the present invention are known and can be purchased on the market, or can be synthesized by or according to methods known in the art.

中间体1-4的制备Preparation of intermediate 1-4

第一步：化合物1-2的制备
Step 1: Preparation of compound 1-2

室温下，向6-溴酞嗪-1(2H)-酮(1-1)(5.0g，22.2mmol，1.0eq.)，叔丁基哌嗪-1-羧酸酯(8.0g，44.4mmol，2.0eq.)，NaOtBu(4.3g，44.4mmol，2.0eq.)的甲苯(60mL)溶液中加入Pd2(dba)3(0.8g，0.89mmol，0.04eq.)和RuPhos(0.85g，1.78mmol，0.08eq.)。将反应混合物在110℃下在N2气氛下搅拌5小时。冷至室温，用饱和NH4Cl水溶液(60mL)淬灭混合物，然后用EA(120mL)萃取混合物。将有机层浓缩至约60mL并形成沉淀。过滤混合物，用DCM(15mL)和MeOH(5mL)洗涤滤饼，干燥，得到4-(1-氧代-1，2-二氢酞嗪-6-基)哌嗪-1-羧酸叔丁酯(1-2)(5.5g)。MS:m/z＝331[M+H]+。Pd 2 (dba) 3 (0.8 g, 0.89 mmol, 0.04 eq.) and RuPhos (0.85 g, 1.78 mmol, 0.08 eq.) were added to a solution of 6 -bromophthalazin- 1 (2H)-one (1-1) (5.0 g, 22.2 mmol, 1.0 eq.), tert-butylpiperazine-1-carboxylate (8.0 g, 44.4 mmol, 2.0 eq.) and NaO t Bu (4.3 g, 44.4 mmol, 2.0 eq.) in toluene (60 mL) at room temperature. The reaction mixture was stirred at 110° C. under N 2 atmosphere for 5 hours. After cooling to room temperature, the mixture was quenched with saturated NH 4 Cl aqueous solution (60 mL), and then extracted with EA (120 mL). The organic layer was concentrated to about 60 mL and a precipitate was formed. The mixture was filtered, and the filter cake was washed with DCM (15 mL) and MeOH (5 mL), and dried to give tert-butyl 4-(1-oxo-1,2-dihydrophthalazin-6-yl)piperazine-1-carboxylate (1-2) (5.5 g). MS: m/z=331 [M+H] + .

第二步：化合物1-3的制备
Step 2: Preparation of Compound 1-3

室温下，向4-(1-氧代-1，2-二氢酞嗪-6-基)哌嗪-1-羧酸叔丁酯(1-2)(0.2g，0.61mmol，1.0eq.)的DMF(5mL)溶液中加入NaH(60％ Wt,60mg,1.53mmol,2.5eq.)。在35℃下搅拌30分钟后，将3-溴哌啶-2，6-二酮(174mg，0.92mmol，1.5eq.)在2mL DMF中的溶液滴加到混合物中，并继续在35℃下搅拌4小时。冷至室温，用H2O(10mL)稀释，并用EA(10mL*2)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物经反相色谱(HCOOH缓冲液)纯化，得到4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-羧酸叔丁酯(1-3)(95mg)。MS:m/z＝442[M+H]+。To a solution of tert-butyl 4-(1-oxo-1,2-dihydrophthalazin-6-yl)piperazine-1-carboxylate (1-2) (0.2 g, 0.61 mmol, 1.0 eq.) in DMF (5 mL) was added NaH (60% Wt, 60 mg, 1.53 mmol, 2.5 eq.) at room temperature. After stirring at 35°C for 30 minutes, a solution of 3-bromopiperidine-2,6-dione (174 mg, 0.92 mmol, 1.5 eq.) in 2 mL of DMF was added dropwise to the mixture, and stirring was continued at 35°C for 4 hours. The mixture was cooled to room temperature, diluted with H 2 O (10 mL), and extracted with EA (10 mL*2). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (HCOOH buffer) to give tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazine-1-carboxylate (1-3) (95 mg). MS: m/z = 442 [M+H] + .

第三步：化合物1-4的制备
Step 3: Preparation of Compound 1-4

室温下，向4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-羧酸叔丁酯(1-3)(95mg,0.22mmol)的EA(2毫升)溶液中加入HCl(5mL,4M in EA)。搅拌1小时后，浓缩得到琥珀色固体的3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮盐酸盐(1-4)(95mg)。MS:m/z＝342[M+H]+。At room temperature, HCl (5 mL, 4 M in EA) was added to a solution of tert-butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazine-1-carboxylate (1-3) (95 mg, 0.22 mmol) in EA (2 mL). After stirring for 1 hour, the mixture was concentrated to give 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione hydrochloride (1-4) (95 mg) as an amber solid. MS: m/z=342[M+H] + .

中间体2-3的制备
Preparation of intermediate 2-3

第一步：化合物9-1的制备Step 1: Preparation of compound 9-1

室温下，向4-氟苯甲酸叔丁酯(5.0g，25.5mmol，1.0eq.)和哌啶-4-基甲醇(5.9g，51.0mmol，2.0eq.)的DMF(40mL)溶液中加入K2CO3(8.8g，51.0mmol，2.0eq.)。在118℃搅拌16小时后，冷至室温，加入H2O(60mL)和EtOAc(200mL)。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物用硅胶柱纯化PE/EA(50/1至1/2)，得到4-(4-(羟甲基)哌啶-1-基)苯甲酸叔丁酯(9-1)(5.9g)。MS:m/z＝292[M+H]+ To a solution of tert-butyl 4-fluorobenzoate (5.0 g, 25.5 mmol, 1.0 eq.) and piperidin-4-ylmethanol (5.9 g, 51.0 mmol, 2.0 eq.) in DMF (40 mL) was added K 2 CO 3 (8.8 g, 51.0 mmol, 2.0 eq.) at room temperature. After stirring at 118° C. for 16 hours, the mixture was cooled to room temperature, and H 2 O (60 mL) and EtOAc (200 mL) were added. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column PE/EA (50/1 to 1/2) to give tert-butyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate (9-1) (5.9 g). MS: m/z=292[M+H] +

第一步：化合物2-3的制备Step 1: Preparation of compound 2-3

向化合物9-1(2.9g，10mmol)和Et3N(3g，30mmol，3eq.)的DCM(50mL)溶液中，在0℃下分批加入MsCl(1.72g，15mmol，1.5eq.)。将反应混合物在室温下搅拌2h，用H2O(100mL)稀释并用DCM(50mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，并减压浓缩。残余物通过硅胶柱(PE：EA＝5：1)纯化，得到4-(4-(((甲基磺酰基)氧基)甲基)哌啶-1-基)苯甲酸叔丁酯2-3(3.24g)。MS:m/z＝370[M+H]+。To a solution of compound 9-1 (2.9 g, 10 mmol) and Et 3 N (3 g, 30 mmol, 3 eq.) in DCM (50 mL) was added MsCl (1.72 g, 15 mmol, 1.5 eq.) in portions at 0°C. The reaction mixture was stirred at room temperature for 2 h, diluted with H 2 O (100 mL) and extracted with DCM (50 mL*3). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column (PE:EA=5:1) to give tert-butyl 4-(4-(((methylsulfonyl)oxy)methyl)piperidin-1-yl)benzoate 2-3 (3.24 g). MS: m/z=370[M+H] + .

中间体5-4的制备Preparation of intermediate 5-4

第一步：化合物5-3的制备
Step 1: Preparation of compound 5-3

室温下，向(R)-3-((甲苯磺酰氧基)甲基)吡咯烷-1-羧酸叔丁酯(1.50g、4.22mmol、1.0eq.)、2-(2，6-二氧代哌啶-3-基)-5-羟基异吲哚啉-1，3-二酮(1.16g、4.22mmol、1.0eq.)的DMF(15mL)溶液中，加入K2CO3(1.75g，12.66mmol，3.0eq.)和NaI(63mg，0.422mmol，0.1eq.)。在80℃下搅拌3小时后，冷至室温，加入H2O(100mL)和EA(100mL)，并用EA(3x 100mL)萃取水 层。将有机层干燥并蒸发。残余物通过用PE/EA(1：1至1：2)洗脱的硅胶上色谱纯化，得到叔丁基(3R)-3-((((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-羧酸酯(5-3)(1.16g)。MS:m/z＝457[M+H]+ To a solution of (R)-tert-butyl 3-((tosyloxy)methyl)pyrrolidine-1-carboxylate (1.50 g, 4.22 mmol, 1.0 eq.) and 2-(2,6-dioxopiperidin-3-yl)-5-hydroxyisoindoline-1,3-dione (1.16 g, 4.22 mmol, 1.0 eq.) in DMF (15 mL) at room temperature were added K 2 CO 3 (1.75 g, 12.66 mmol, 3.0 eq.) and NaI (63 mg, 0.422 mmol, 0.1 eq.). After stirring at 80° C. for 3 hours, the mixture was cooled to room temperature, H 2 O (100 mL) and EA (100 mL) were added, and water was extracted with EA (3×100 mL). The organic layer was dried and evaporated. The residue was purified by chromatography on silica gel eluting with PE/EA (1:1 to 1:2) to give tert-butyl (3R)-3-((((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (5-3) (1.16 g). MS: m/z=457 [M+H] +

第二步：化合物5-4的制备
Step 2: Preparation of compound 5-4

室温下，向叔丁基(3R)-3-((((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-羧酸酯(5-3)(200毫克，0.437毫摩尔，1.0eq.)的EA溶液中加入4M HCl in ethyl acetate(1mL,4mmol)。室温下搅拌2小时,浓缩得到2-(2，6-二氧代哌啶-3-基)-5-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-1，3-二酮盐酸盐(5-4)(160毫克)。MS:m/z＝357[M+H]+ To a solution of tert-butyl (3R)-3-((((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (5-3) (200 mg, 0.437 mmol, 1.0 eq.) in EA was added 4M HCl in ethyl acetate (1 mL, 4 mmol) at room temperature. The mixture was stirred at room temperature for 2 hours and concentrated to give 2-(2,6-dioxopiperidin-3-yl)-5-(((R)-pyrrolidin-3-yl)methoxy)isoindolin-1,3-dione hydrochloride (5-4) (160 mg). MS: m/z=357 [M+H] +

中间体5-5的制备
Preparation of intermediate 5-5

第一步：化合物5-5-1的制备Step 1: Preparation of compound 5-5-1

室温下，向8-溴-5-氟喹啉(2克，8.85毫摩尔，1.0等量,Zn(CN)2(2.07克，17.7毫摩尔，2.0当量) 的DMF(20毫升)溶液中加入RuPhos Pd G3(300毫克，0.35毫摩尔，0.04当量)。将反应混合物在N2气氛下在100℃搅拌16小时。反应完成后，用H2O(200mL)稀释，并用EA(50mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，真空过滤浓缩，硅胶柱纯化(PE：EA＝2：1)，得到5-氟喹啉-8-甲腈(5-5-1)(1.4g)。MS:m/z＝173[M+H]+ At room temperature, 8-bromo-5-fluoroquinoline (2 g, 8.85 mmol, 1.0 equiv., Zn(CN) 2 (2.07 g, 17.7 mmol, 2.0 equiv.) RuPhos Pd G3 (300 mg, 0.35 mmol, 0.04 eq) was added to a DMF (20 mL) solution. The reaction mixture was stirred at 100 °C for 16 hours under N2 atmosphere. After the reaction was completed, it was diluted with H2O (200 mL) and extracted with EA (50 mL*3). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by silica gel column (PE:EA=2:1) to obtain 5-fluoroquinoline-8-carbonitrile (5-5-1) (1.4 g). MS: m/z=173[M+H] +

第二步：化合物5-5-2的制备Step 2: Preparation of compound 5-5-2

室温下，向5-氟喹啉-8-甲腈(5-5-1)(200毫克，1.16毫摩尔，1.0当量)，叔丁基((1r，4r)-4-羟基环己基)氨基甲酸叔丁酯(274毫克，1.28毫摩尔，1.1当量)的MeCN(2毫升)溶液中加入Cs2CO3(755毫克，2.32毫摩尔，2.0当量)。将反应混合物在90℃下搅拌16小时。反应完成后，冷至室温，用H2O(20mL)稀释，并用EA(10mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，真空过滤浓缩，硅胶柱纯化(MeCN：H2O/9:1)，得到((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)氨基甲酸叔丁酯(5-5-2)(220毫克)。MS:m/z＝367[M+H]+ Cs 2 CO 3 (755 mg, 2.32 mmol, 2.0 eq) was added to a solution of 5-fluoroquinoline-8-carbonitrile ( 5-5-1 ) (200 mg, 1.16 mmol, 1.0 eq) and tert-butyl ((1r, 4r)-4-hydroxycyclohexyl)carbamate (274 mg, 1.28 mmol, 1.1 eq) in MeCN (2 ml) at room temperature. The reaction mixture was stirred at 90° C. for 16 hours. After completion of the reaction, the mixture was cooled to room temperature, diluted with H 2 O (20 mL), and extracted with EA (10 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by silica gel column (MeCN: H2O /9:1) to obtain tert-butyl ((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)carbamate (5-5-2) (220 mg). MS: m/z=367[M+H] +

第三步：化合物1-7的制备Step 3: Preparation of Compound 1-7

室温下,向((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)氨基甲酸叔丁酯(5-5-2)(3g，8.17mmol，1.0eq.)的EA(10mL)溶液中加入4M HCl的EA(30mL)溶液。搅拌16小时后，将反应液真空下浓缩，得到5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(2.9克，粗品)。MS:m/z＝267[M+H]+ At room temperature, a 4M HCl solution in EA (30 mL) was added to a solution of tert-butyl ((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)carbamate (5-5-2) (3 g, 8.17 mmol, 1.0 eq.) in EA (10 mL). After stirring for 16 hours, the reaction solution was concentrated under vacuum to give 5-(((1r, 4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (2.9 g, crude product). MS: m/z = 267 [M+H] +

第四步：化合物1-5的制备Step 4: Preparation of Compound 1-5

室温下，将4-(4-(羟甲基)哌啶-1-基)苯甲酸叔丁酯(5g，17.18mmol，1.0eq.)，IBX(5.75g，20.61mmol，1.2eq.)在DMSO(50mL)溶液中混合。将反应混合物在60℃下搅拌2小时。反应完成后，冷至室温，用H2O(500mL)稀释，并用EA(100mL*3)萃取。合并有机层用饱和食盐水洗涤，无水硫酸钠干燥，真空过滤浓缩，硅胶柱(PE：EA＝1：1)，得到4-(4-甲酰基哌啶-1-基)苯甲酸叔丁酯(1-5)(4.2g)。MS:m/z＝289[M+H]+ At room temperature, tert-butyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate (5 g, 17.18 mmol, 1.0 eq.) and IBX (5.75 g, 20.61 mmol, 1.2 eq.) were mixed in a DMSO (50 mL) solution. The reaction mixture was stirred at 60°C for 2 hours. After the reaction was completed, it was cooled to room temperature, diluted with H 2 O (500 mL), and extracted with EA (100 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, vacuum filtered and concentrated, and silica gel column (PE:EA=1:1) was used to obtain tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (1-5) (4.2 g). MS: m/z=289[M+H] +

第五步：化合物5-5-5的制备Step 5: Preparation of compound 5-5-5

室温下,向4-(4-甲酰基哌啶-1-基)苯甲酸叔丁酯(1-5)(2克，6.92毫摩尔，1.0当量)的DCM(50毫升)溶液中加入TFA(5毫升)。搅拌16小时后，将反应液混合物在真空下浓缩，得到4-(4-(1,3-二氧杂环戊-2-基)哌啶-1-基)苯甲酸叔丁酯(5-5-5)(1.9克，粗品)。MS:m/z＝233[M+H]+ At room temperature, TFA (5 mL) was added to a solution of tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (1-5) (2 g, 6.92 mmol, 1.0 eq) in DCM (50 mL). After stirring for 16 hours, the reaction mixture was concentrated under vacuum to give tert-butyl 4-(4-(1,3-dioxolan-2-yl)piperidin-1-yl)benzoate (5-5-5) (1.9 g, crude). MS: m/z = 233 [M+H] +

第六步：化合物5-5-6的制备Step 6: Preparation of compound 5-5-6

室温下，向4(4-(1，3-二氧杂环戊-2-基)哌啶-1-基)苯甲酸叔丁酯(5-5-5)(2g，8.58mmol，1.0当量)，乙二醇(800μL，12.87毫摩尔，1.5等量)的甲苯(20mL)溶液中，加入TsOH(40mg，0.25毫摩尔，0.03当量)。将反应混合物在100℃搅拌1小时。反应完成后，将混合物在真空下浓缩并用硅胶柱纯化(MeCN：H2O/55％:45％)，得到4-(4-(1，3-二氧戊环-2-基)哌啶-1-基)苯甲酸(5-5-6)(1.29克)。MS:m/z＝277[M+H]+ To a solution of tert-butyl 4-(4-(1,3-dioxolan-2-yl)piperidin-1-yl)benzoate (5-5-5) (2 g, 8.58 mmol, 1.0 equiv), ethylene glycol (800 μL, 12.87 mmol, 1.5 equiv) in toluene (20 mL) at room temperature, TsOH (40 mg, 0.25 mmol, 0.03 equiv) was added. The reaction mixture was stirred at 100°C for 1 hour. After the reaction was complete, the mixture was concentrated under vacuum and purified with a silica gel column (MeCN:H 2 O/55%:45%) to give 4-(4-(1,3-dioxolan-2-yl)piperidin-1-yl)benzoic acid (5-5-6) (1.29 g). MS: m/z=277[M+H] +

第七步：化合物5-5-7的制备Step 7: Preparation of compound 5-5-7

室温下,向4-(4-(1，3-二氧戊环-2-基)哌啶-1-基)苯甲酸(5-5-6)(1.29g，4.65mmol，1.0 eq.)、5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(1.25g、4.65mmol、1.0当量)，HATU(2.13g、5.58mmol、1.2eq.)的DMF(25mL)溶液中，加入DIEA(4mL、23.2mmol、5.0当量)。将反应混合物在80℃搅拌6小时，冷至室温，过滤混合物，用DCM(5mL)洗涤滤饼。滤液在真空下浓缩，得到4-(4-(1，3-二氧戊环-2-基)哌啶-1-基)-N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)苯甲酰胺(5-5-7)(2.0g)。MS:m/z＝526[M+H]+ At room temperature, 4-(4-(1,3-dioxolan-2-yl)piperidin-1-yl)benzoic acid (5-5-6) (1.29 g, 4.65 mmol, 1.0 eq.), 5-(((1r, 4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (1.25 g, 4.65 mmol, 1.0 eq.), HATU (2.13 g, 5.58 mmol, 1.2 eq.) in DMF (25 mL), DIEA (4 mL, 23.2 mmol, 5.0 eq.) was added. The reaction mixture was stirred at 80°C for 6 hours, cooled to room temperature, the mixture was filtered, and the filter cake was washed with DCM (5 mL). The filtrate was concentrated under vacuum to give 4-(4-(1,3-dioxolan-2-yl)piperidin-1-yl)-N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)benzamide (5-5-7) (2.0 g). MS: m/z=526[M+H] +

第八步：化合物5-5的制备Step 8: Preparation of compound 5-5

室温下，向4-(4-(1，3-二氧戊环-2-基)哌啶-1-基)-N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)苯甲酰胺(5-5-7)(2g，3.8mmol，1.0eq.)的MeCN(10mL)和H2O(50mL)溶液中加入12N HCl(5mL，28.5mmol，7.5eq.)。将反应混合物在60℃搅拌16小时后，柱层析纯化(MeCN：H2O＝75％)，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(1.8克)。MS:m/z＝482[M+H]+ To a solution of 4-(4-(1,3-dioxolan-2-yl)piperidin-1-yl)-N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)benzamide (5-5-7) (2 g, 3.8 mmol, 1.0 eq.) in MeCN (10 mL) and H 2 O (50 mL) was added 12N HCl (5 mL, 28.5 mmol, 7.5 eq.) at room temperature. The reaction mixture was stirred at 60°C for 16 hours and then purified by column chromatography (MeCN:H 2 O=75%) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (1.8 g). MS: m/z=482[M+H] +

中间体21-6的制备
Preparation of Intermediate 21-6

第一步：化合物21-2的制备Step 1: Preparation of compound 21-2

室温下，向3-(2，6-双(苄氧基)吡啶-3-基)-7-溴-1-甲基-1H-吲唑(21-1)(100毫克，0.2毫摩尔，1.0eq.)的溶液中加入B2Pin2(60毫克，0.24毫摩尔，1.2eq.)的1,4-dioxane(5mL)溶液中加入KOAc(40毫克，0.4毫摩尔，2.0eq.)和Pd(dppf)Cl2(10毫克，0.01毫摩尔，0.05eq.)。在85℃搅拌16小时后，冷至室温，加入用H2O(20mL)，并用EA(5mL*2)萃取。合并有机层饱和食盐水洗涤，无水硫酸钠干燥，过滤，真空浓缩。残余物用硅胶色谱(PE：EA＝50：1)纯化，得到3-(2，6-双(苄氧基)吡啶-3-基)-1-甲基-7-(4，4，5，5-四甲基-1，3，2-二氧杂硼烷-2-基)-1H-吲唑(21-2)(100mg)。MS:m/z＝547[M+H]+ To a solution of 3-(2,6-bis(benzyloxy)pyridin-3-yl)-7-bromo-1-methyl-1H-indazole (21-1) (100 mg, 0.2 mmol, 1.0 eq.) was added a solution of B 2 Pin 2 (60 mg, 0.24 mmol, 1.2 eq.) in 1,4-dioxane (5 mL) at room temperature, KOAc (40 mg, 0.4 mmol, 2.0 eq.) and Pd(dppf)Cl 2 (10 mg, 0.01 mmol, 0.05 eq.) were added. After stirring at 85°C for 16 hours, the mixture was cooled to room temperature, H 2 O (20 mL) was added, and the mixture was extracted with EA (5 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (PE:EA=50:1) to give 3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (21-2) (100 mg). MS: m/z=547 [M+H] +

第二步：化合物21-3的制备Step 2: Preparation of compound 21-3

室温下，向3-(2，6-双(苄氧基)吡啶-3-基)-1-甲基-7-(4，4，5，5-四甲基-1，3，2-二氧杂硼烷-2-基)-1H-吲唑(21-2)(100毫克，0.18毫摩尔，1.0eq.)的THF(3mL)溶液中加入30％H2O2(100μL，0.9毫摩尔，10.0eq)。搅拌6小时后，浓缩，得到3-(2，6-双(苄氧基)吡啶-3-基)-1-甲基-1H-吲唑-7-醇(21-3)(100毫克)。MS:m/z＝437[M+H]+ To a solution of 3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-indazole (21-2) (100 mg, 0.18 mmol, 1.0 eq.) in THF (3 mL) was added 30% H 2 O 2 (100 μL, 0.9 mmol, 10.0 eq.) at room temperature. After stirring for 6 hours, the mixture was concentrated to give 3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-7-ol (21-3) (100 mg). MS: m/z=437[M+H] +

第三步：化合物21-4的制备Step 3: Preparation of compound 21-4

室温下，向3-(2，6-双(苄氧基)吡啶-3-基)-1-甲基-1H-吲唑-7-醇(21-3)(50毫克，0.23毫摩 尔，1.0eq.)和叔丁基(R)-3-((甲苯磺酰氧基)甲基)吡咯烷-1-羧酸酯(97毫克，0.27毫摩尔，1.2eq.)的DMF(2mL)溶液中加入Cs2CO3(150毫克，0.46毫摩尔，2.0eq.)，在80℃下搅拌1小时后，冷至室温，浓缩，并在硅胶柱纯化(PE：EA＝2：1)，得到((3-(2，6-双(苄氧基)吡啶-3-基)-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-羧酸酯(21-4)(100毫克)。MS:m/z＝620[M+H]+ At room temperature, 3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-7-ol (21-3) (50 mg, 0.23 mmol) was added to To a solution of tert-butyl (R)-3-((tosyloxy)methyl)pyrrolidine-1-carboxylate (97 mg, 0.27 mmol, 1.2 eq.) in DMF (2 mL) was added Cs 2 CO 3 (150 mg, 0.46 mmol, 2.0 eq.), stirred at 80°C for 1 hour, cooled to room temperature, concentrated, and purified on a silica gel column (PE:EA=2:1) to give ((3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (21-4) (100 mg). MS: m/z=620[M+H] +

第四步：化合物21-5的制备Step 4: Preparation of compound 21-5

室温下,向((3-(2，6-双(苄氧基)吡啶-3-基)-1-甲基-1H-吲唑-7-氧基)甲基)吡咯烷-1-羧酸叔丁酯(21-4)(100毫克，0.16毫摩尔，1.0eq.)，10％Pd/C(20毫克，20％w.t)在EA(5mL)中的混合物在H2气氛下室温搅拌16小时。过滤并在真空下浓缩，并用硅胶柱纯化(PE：EA＝1：1)，得到(3-(2，6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-羧酸酯(21-5)(40mg)。MS:m/z＝442[M+H]+ At room temperature, a mixture of ((3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester (21-4) (100 mg, 0.16 mmol, 1.0 eq.), 10% Pd/C (20 mg, 20% wt) in EA (5 mL) was stirred at room temperature under H 2 atmosphere for 16 hours. Filtered and concentrated under vacuum, and purified by silica gel column (PE:EA=1:1) to give (3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (21-5) (40 mg). MS: m/z=442[M+H] +

第五步：化合物21-6的制备Step 5: Preparation of compound 21-6

向((3-(2，6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-羧酸叔丁酯(21-5)(40mg，0.09mmol，1.0eq.)的EA(2mL)溶液中加入HCl/EA(1mL)。室温搅拌16小时后，浓缩，得到3-(1-甲基-7-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2，6-二酮(21-6)(40毫克)。MS:m/z＝342[M+H]+ To a solution of tert-butyl ((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (21-5) (40 mg, 0.09 mmol, 1.0 eq.) in EA (2 mL) was added HCl/EA (1 mL). After stirring at room temperature for 16 hours, the mixture was concentrated to give 3-(1-methyl-7-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione (21-6) (40 mg). MS: m/z=342[M+H] +

中间体58-1的制备
Preparation of Intermediate 58-1

向4-溴苯甲酸叔丁酯(200mg，0.77mmol，1.0eq.)、(2R,4R)-2-甲基哌啶-4-甲酸甲酯(120mg，0.77mmol，1.0eq.)、Cs2CO3(750mg，2.31mmol，3.0eq.)于甲苯(10mL)中的溶液中添加RuPhosPdG3(100mg，0.15mmol，0.2eq.)、RuPhos(70mg，0.15mmol，0.2eq.)。将反应混合物在110℃下在N2气氛下搅拌16小时。在反应完成后，将混合物用H2O(100mL)稀释，并用EA(50mL×2)萃取。将合并的有机层用盐水洗涤，经无水硫酸钠干燥，过滤并真空浓缩。将残余物通过硅胶色谱用PE:EA＝10:1洗脱纯化，得到呈黄色固体状的(2R,4R)-1-(4-(叔丁氧基羰基)苯基)-2-甲基哌啶-4-甲酸甲酯(58-1-1)(230mg，88％)。MS:m/z＝277[M+H-56]+。To a solution of tert-butyl 4-bromobenzoate (200 mg, 0.77 mmol, 1.0 eq.), methyl (2R, 4R)-2-methylpiperidine-4-carboxylate (120 mg, 0.77 mmol, 1.0 eq.), Cs 2 CO 3 (750 mg, 2.31 mmol, 3.0 eq.) in toluene (10 mL) were added RuPhosPdG3 (100 mg, 0.15 mmol, 0.2 eq.), RuPhos (70 mg, 0.15 mmol, 0.2 eq.). The reaction mixture was stirred at 110° C. under N 2 atmosphere for 16 hours. After completion of the reaction, the mixture was diluted with H 2 O (100 mL) and extracted with EA (50 mL×2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo. The residue was purified by silica gel chromatography eluting with PE:EA=10:1 to obtain methyl (2R,4R)-1-(4-(tert-butoxycarbonyl)phenyl)-2-methylpiperidine-4-carboxylate (58-1-1) (230 mg, 88%) as a yellow solid. MS: m/z=277 [M+H-56] + .

向(2R,4R)-1-(4-(叔丁氧基羰基)苯基)-2-甲基哌啶-4-甲酸甲酯(58-1-1)(230mg，0.69mmol，1.0eq.)于DCM(4mL)中的溶液中添加TFA(2mL)。将反应混合物在室温下搅拌16小时。在反应完成后，真空浓缩混合物，得到呈黄色油状的4-((2R,4R)-4-(甲氧基羰基)-2-甲基哌啶-1-基)苯甲酸(58-1)(250mg，粗物质)。MS:m/z＝277[M+H]+。To a solution of (2R, 4R)-1-(4-(tert-butoxycarbonyl)phenyl)-2-methylpiperidine-4-carboxylic acid methyl ester (58-1-1) (230 mg, 0.69 mmol, 1.0 eq.) in DCM (4 mL) was added TFA (2 mL). The reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete, the mixture was concentrated in vacuo to give 4-((2R, 4R)-4-(methoxycarbonyl)-2-methylpiperidin-1-yl)benzoic acid (58-1) (250 mg, crude material) as a yellow oil. MS: m/z=277[M+H] + .

中间体67-6的制备
Preparation of Intermediate 67-6

向4-((2R,4R)-4-(甲氧基羰基)-2-甲基哌啶-1-基)苯甲酸(58-1)(100mg，0.36mmol，1.0eq.)、5-((1r,3r)-3-氨基-2,2,4,4-四甲基环丁氧基)喹啉-8-甲腈(67-3)(106mg，0.36mmol，1.0eq.)、HATU(164mg，0.43mmol，1.2eq.)于DMF(2mL)中的溶液中添加DIEA(200μL，1.08mmol，3.0eq.)。将反应混合物在室温下搅拌16小时。在反应完成后，将混合物通过色谱用MeCN:H2O＝90％纯化，得到呈黄色固体状的(2R,4R)-1-(4-(((1r,3R)-3-((8-氰基喹啉-5-基)氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基)苯基)-2-甲基哌啶-4-甲酸甲酯(67-4)(150mg，75％)。MS:m/z＝554[M+H]+。DIEA (200 μL, 1.08 mmol, 3.0 eq.) was added to a solution of 4-((2R,4R)-4-(methoxycarbonyl)-2-methylpiperidin-1-yl)benzoic acid (58-1) (100 mg, 0.36 mmol, 1.0 eq.), 5-((1r,3r)-3-amino-2,2,4,4-tetramethylcyclobutyloxy)quinoline-8-carbonitrile (67-3) (106 mg, 0.36 mmol, 1.0 eq.), HATU (164 mg, 0.43 mmol, 1.2 eq.) in DMF (2 mL). The reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was purified by chromatography with MeCN:H 2 O=90% to give methyl (2R,4R)-1-(4-(((1r,3R)-3-((8-cyanoquinolin-5-yl)oxy)-2,2,4,4-tetramethylcyclobutyl)carbamoyl)phenyl)-2-methylpiperidine-4-carboxylate (67-4) (150 mg, 75%) as a yellow solid. MS: m/z=554 [M+H] + .

向(2R,4R)-1-(4-(((1r,3R)-3-((8-氰基喹啉-5-基)氧基)-2,2,4,4-四甲基环丁基)氨基甲酰基)苯基)-2-甲基哌啶-4-甲酸甲酯(67-4)(140mg，0.25mmol，1.0eq.)、LiBH4(55mg，2.5mmol，10eq.)于THF(10mL)中的溶液中。将反应混合物在室温下搅拌16小时。在反应完成后，将混合物通过色谱用MeCN:H2O＝90％纯化，得到呈白色固体状的N-((1r,3R)-3-((8-氰基喹啉-5-基)氧基)-2,2,4,4-四甲基环丁基)-4-((2R,4R)-4-(羟甲基)-2-甲基哌啶-1-基)苯甲酰胺(67-5)(130mg，98％)。MS:m/z＝526[M+H]+。To a solution of methyl (2R,4R)-1-(4-(((1r,3R)-3-((8-cyanoquinolin-5-yl)oxy)-2,2,4,4-tetramethylcyclobutyl)carbamoyl)phenyl)-2-methylpiperidine-4-carboxylate (67-4) (140 mg, 0.25 mmol, 1.0 eq.), LiBH4 (55 mg, 2.5 mmol, 10 eq.) in THF (10 mL) was added. The reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was purified by chromatography with MeCN:H 2 O=90% to give N-((1r,3R)-3-((8-cyanoquinolin-5-yl)oxy)-2,2,4,4-tetramethylcyclobutyl)-4-((2R,4R)-4-(hydroxymethyl)-2-methylpiperidin-1-yl)benzamide (67-5) (130 mg, 98%) as a white solid. MS: m/z=526[M+H] + .

向N-((1r,3R)-3-((8-氰基喹啉-5-基)氧基)-2,2,4,4-四甲基环丁基)-4-((2R,4R)-4-(羟甲基)-2-甲基哌啶-1-基)苯甲酰胺(67-5)(130mg，0.24mmol，1.0eq.)于DCM(5mL)中的溶液中添加DMP(135mg，0.32mmol，1.2eq.)。将反应混合物在室温下搅拌16小时。在反应完成后，将混合物用饱和NaS2O3(50mL)稀释，并分别用饱和NaHCO3(50mL)和H2O(50mL)洗涤，水层用DCM(10mL)萃取。合并的有机层用盐水洗涤，经无水硫酸钠干燥，过滤并真空浓缩，得到呈黄色固体状的N-((1r,3R)-3-((8-氰基喹啉-5-基)氧基)-2,2,4,4-四甲基环丁基)-4-((2R,4R)-4-甲酰基-2-甲基哌啶-1-基)苯甲酰胺(67-6)(120mg，粗物质)。MS:m/z＝524[M+H]+。To a solution of N-((1r,3R)-3-((8-cyanoquinolin-5-yl)oxy)-2,2,4,4-tetramethylcyclobutyl)-4-((2R,4R)-4-(hydroxymethyl)-2-methylpiperidin-1-yl)benzamide (67-5) (130 mg, 0.24 mmol, 1.0 eq.) in DCM (5 mL) was added DMP (135 mg, 0.32 mmol, 1.2 eq.). The reaction mixture was stirred at room temperature for 16 hours. After the reaction was completed, the mixture was diluted with saturated NaS 2 O 3 (50 mL) and washed with saturated NaHCO 3 (50 mL) and H 2 O (50 mL), respectively, and the aqueous layer was extracted with DCM (10 mL). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give N-((1r,3R)-3-((8-cyanoquinolin-5-yl)oxy)-2,2,4,4-tetramethylcyclobutyl)-4-((2R,4R)-4-formyl-2-methylpiperidin-1-yl)benzamide (67-6) (120 mg, crude) as a yellow solid. MS: m/z=524 [M+H] + .

实施例1化合物1的制备Example 1 Preparation of Compound 1

第一步：化合物1-6的制备
Step 1: Preparation of Compound 1-6

室温下，向3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮(1-4)(180mg，0.48mmol，1.0eq.)的DCM/MeOH(6mL/4mL)溶液中加入4-(4-甲酰基哌啶-1-基)苯甲酸叔丁酯(1-5)(132mg，0.46mmol，0.96eq.)和TEA(18滴)，搅拌3小时后，加入NaBH(OAc)3(270mg，1.30mmol，2.7eq.)。室温下搅拌1小时，减压浓缩至干。向残留物中在加入EA/H2O(20mL+20mL)，并用EA(20mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤浓缩，得到粗品叔丁基4-(4-((2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸酯(1-6)(280mg，粗品)，直接用于下一步反应。MS:m/z＝615[M+H]+ At room temperature, tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (1-5) (132 mg, 0.46 mmol, 0.96 eq.) and TEA (18 drops) were added to a solution of 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione (1-4) (180 mg, 0.48 mmol, 1.0 eq.) in DCM/MeOH (6 mL/4 mL). After stirring for 3 hours, NaBH(OAc) 3 (270 mg, 1.30 mmol, 2.7 eq.) was added. The mixture was stirred at room temperature for 1 hour and concentrated to dryness under reduced pressure. EA/H 2 O (20 mL+20 mL) was added to the residue and extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give crude tert-butyl 4-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (1-6) (280 mg, crude), which was directly used in the next step. MS: m/z=615[M+H] +

第二步：化合物1的制备
Step 2: Preparation of compound 1

室温下，向4-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(1-6)(90mg，0.15mmol，1.0eq.)的DCM(4mL)溶液中加入TFA(2mL)。室温下搅拌1h后，浓缩。将残留物溶解在DMF(4.5mL)中，并加入5-(((1r，4r)-4-氨基环己氧基)喹啉-8-甲腈(1-7)(56mg，0.19mmol，1.25eq.)，HATU(62mg，0.17mmol，1.1eq.)和DIPEA(193mg，1.5mmol，10.0eq.)。将反应混合物在室温下搅拌1h后，用H2O(20mL)稀释混合物并用EA(20mL*3)萃取。合并有机层，用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。所得残余物用高效液相色谱(HCOOH缓冲液)纯化，得到N N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1-氧代-1,2-二氢邻苯二甲酸-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酰胺(1)(11mg)。MS:m/z＝808.4[M+H]+ To a solution of tert-butyl 4-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (1-6) (90 mg, 0.15 mmol, 1.0 eq.) in DCM (4 mL) was added TFA (2 mL) at room temperature. The mixture was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DMF (4.5 mL), and 5-(((1r, 4r)-4-aminocyclohexyloxy)quinoline-8-carbonitrile (1-7) (56 mg, 0.19 mmol, 1.25 eq.), HATU (62 mg, 0.17 mmol, 1.1 eq.) and DIPEA (193 mg, 1.5 mmol, 10.0 eq.) were added. The reaction mixture was stirred at room temperature for 1 h, then diluted with H 2 O (20 mL) and extracted with EA (20 mL*3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by high performance liquid chromatography (HCOOH buffer) to give N N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalic acid-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (1) (11 mg). MS: m/z=808.4 [M+H] +

1H NMR(400MHz,DMSO-d6)δ10.99(s,1H),9.06(d,J＝1.6Hz,1H),8.64(dd,J＝8.5,1.6Hz,1H),8.28(d,J＝8.5Hz,1H),8.24(s,1H),8.04(d,J＝9.2Hz,1H),7.98(d,J＝7.5Hz,1H),7.74(d,J＝8.8Hz,2H),7.68(dd,J＝8.5,4.3Hz,1H),7.49(dd,J＝9.3,1.6Hz,1H),7.35(d,J＝8.5Hz,1H),7.27–7.22(m,1H),6.93(d,J＝8.8Hz,2H),5.77–5.72(m,1H),4.77–4.68(m,1H),3.93–3.78(m,3H),3.42(br,3H),2.98–2.85(m,1H),2.78(t,J＝11.6Hz,2H),2.54–2.52(m,3H),2.29–2.19(m,4H),1.98–1.92(m,2H),1.81(d,J＝10.3Hz,3H),1.72–1.51(m,5H),1.29–1.20(m,5H). 1 H NMR (400MHz, DMSO-d 6 )δ10.99(s,1H),9.06(d,J＝1.6Hz,1H),8.64(dd,J＝8.5,1.6Hz,1H),8.28(d,J＝8.5Hz,1H),8.24( s,1H),8.04(d,J＝9.2Hz,1H),7.98 (d,J＝7.5Hz,1H),7.74(d,J＝8.8Hz,2H),7.68(dd,J＝8.5,4.3Hz,1H),7.49(dd,J＝9.3,1.6Hz,1H) ,7.35(d,J＝8.5Hz,1H),7.27–7.22( m,1H),6.93(d,J＝8.8Hz,2H),5.77–5.72(m,1H),4.77–4.68(m,1H),3.93–3.78(m,3H),3.42(br,3H) ,2.98–2.85(m,1H),2.78(t,J＝ 11.6Hz,2H),2.54–2.52(m,3H),2.29–2.19(m,4H),1.98–1.92(m,2H),1.81(d,J＝10.3Hz,3H),1.72–1.51(m ,5H),1.29–1.20(m,5H).

实施例2化合物2的制备
Example 2 Preparation of Compound 2

第一步：化合物2-1的制备
Step 1: Preparation of compound 2-1

将4-氟苯甲酸甲酯(2.1g，13.6mmol)，哌嗪-1-羧酸叔丁酯(5.06g，27.2mmol 2.0eq.)和K2CO3(3.75g，27.2mmol 2.0eq.)在DMSO(20mL)中的混合物在120℃下搅拌24h。冷至室温，用H2O(50mL)稀释混合物并用EA(50mL*3)萃取。合并有机层，用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物经硅胶柱(PE：EA＝1：1)纯化，得到4-(4-(甲氧基羰基)苯基)哌嗪-1-羧酸叔丁酯(2-1)(1.74g)。MS:m/z＝321[M+H]+ A mixture of methyl 4-fluorobenzoate (2.1 g, 13.6 mmol), tert-butyl piperazine-1-carboxylate (5.06 g, 27.2 mmol 2.0 eq.) and K 2 CO 3 (3.75 g, 27.2 mmol 2.0 eq.) in DMSO (20 mL) was stirred at 120°C for 24 h. After cooling to room temperature, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL*3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column (PE:EA=1:1) to give tert-butyl 4-(4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate (2-1) (1.74 g). MS: m/z=321[M+H] +

第二步：化合物2-2的制备
Step 2: Preparation of compound 2-2

室温下，向4-(4-(甲氧基羰基)苯基)哌嗪-1-羧酸叔丁酯(2-1)(1.74g，5.4mmol)的二氧六环(20mL)的溶液中加入HCl(5mL,4M in dioxane)。室温下搅拌6h后，将溶液浓缩，得到琥珀色固体的4-(哌嗪-1-基)苯甲酸甲酯(2-2)(1.1克)。MS:m/z＝221[M+H]+ At room temperature, HCl (5 mL, 4 M in dioxane) was added to a solution of tert-butyl 4-(4-(methoxycarbonyl)phenyl)piperazine-1-carboxylate (2-1) (1.74 g, 5.4 mmol) in dioxane (20 mL). After stirring at room temperature for 6 h, the solution was concentrated to give methyl 4-(piperazin-1-yl)benzoate (2-2) (1.1 g) as an amber solid. MS: m/z = 221 [M+H] +

第三步：化合物2-4的制备Step 3: Preparation of Compound 2-4

室温下，将4-(哌嗪-1-基)苯甲酸甲酯(2-2)(1.0g，4.5mmol)、4-(4-((((甲基磺酰基)氧)甲基)哌啶-1-基)苯甲酸叔丁酯(2-3)(1.66g，4.5mmol，1.0eq.)和Et3N(1.36克，13.5mmol，3.0eq.)在乙腈(20毫升)中的化合物在80℃搅拌12h。冷至室温，加H2O(50mL)稀释并用EA(50mL*3)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物经硅胶柱(PE：EA＝1：1)纯化，得到4-(4-((4-(甲氧基羰基)苯基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(2-4)(500mg)。MS:m/z＝494[M+H]+A mixture of methyl 4-(piperazin-1-yl)benzoate (2-2) (1.0 g, 4.5 mmol), tert-butyl 4-(4-((((methylsulfonyl)oxy)methyl)piperidin-1-yl)benzoate (2-3) (1.66 g, 4.5 mmol, 1.0 eq.) and Et 3 N (1.36 g, 13.5 mmol, 3.0 eq.) in acetonitrile (20 mL) was stirred at 80°C for 12 h at room temperature. The mixture was cooled to room temperature, diluted with H 2 O (50 mL) and extracted with EA (50 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column (PE:EA=1:1) to give tert-butyl 4-(4-((4-(methoxycarbonyl)phenyl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (2-4) (500 mg). MS: m/z = 494 [M + H] +

第四步：化合物2-5的制备
Step 4: Preparation of compound 2-5

室温下，向4-(4-(4-((4-(4-((甲氧基羰基)苯基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸叔 丁酯(2-4)(500mg 1.0mmol)在20mL甲醇：THF：H2O(3：1：1)的溶液中，加入LiOH(96mg 4.0mmol 4.0eq.)。在45℃搅拌12h后，冷至室温，用4N HCl(2mL)中和混合物并用EA(20mL*3)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩，得到粗品4-(4-((4-(叔丁氧羰基)苯基)哌啶-4-基)甲基)哌嗪-1-基)苯甲酸。然后将上述粗品溶解在DMF(10mL)中，加入3-氨基哌啶-2，6-二酮(140mg，1.1mmol，1.1eq.)，HATU(761mg，2.0mmol，2.0eq.)和DIPEA(516mg，4.0mmol，4.0eq.)。室温下搅拌4h后，加日H2O(100mL)，并用EA(100mL*3)萃取。合并有机层用饱和食盐水洗涤，无水硫酸钠干燥，过滤，减压浓缩。残留物通过硅胶柱(PE:EA＝1∶5)纯化，得到白色固体的4-(4-(4-〔4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(2-5)(220mg)。MS:m/z＝590[M+H]+ At room temperature, tert-4-(4-(4-((4-(4-((methoxycarbonyl)phenyl)piperazin-1-yl)methyl)piperidin-1-yl)benzoic acid was added. To a solution of butyl ester (2-4) (500 mg 1.0 mmol) in 20 mL of methanol:THF:H 2 O (3:1:1), LiOH (96 mg 4.0 mmol 4.0 eq.) was added. After stirring at 45°C for 12 h, the mixture was cooled to room temperature, neutralized with 4N HCl (2 mL) and extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude 4-(4-((4-(tert-butyloxycarbonyl)phenyl)piperidin-4-yl)methyl)piperazin-1-yl)benzoic acid. Then the crude product was dissolved in DMF (10 mL), and 3-aminopiperidine-2,6-dione (140 mg, 1.1 mmol, 1.1 eq.), HATU (761 mg, 2.0 mmol, 2.0 eq.) and DIPEA (516 mg, 4.0 mmol, 4.0 eq.) were added. After stirring at room temperature for 4 h, H 2 O (100 mL) was added and extracted with EA (100 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column (PE:EA=1:5) to obtain tert-butyl 4-(4-(4-[4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenyl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (2-5) (220 mg) as a white solid. MS: m/z=590[M+H] +

第五步：化合物2的制备Step 5: Preparation of Compound 2

室温下，向4-((4-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(2-5)(170mg，0.29mmol)的DCM(10mL)中的溶液中加入TFA(5mL)。室温下搅拌4h后，浓缩。将残余物溶解在DMF(20mL)中，并加入5-(((1r，4r)-4-氨基环己氧基)喹啉-8-甲腈(1-7)(310mg，1.16mmol，4.0eq.)，HATU(221mg，0.58毫摩尔，2.0eq.)和DIPEA(150毫克，1.16毫摩尔，4.0eq.)。室温下搅拌4h后，加H2O(50mL)，并用EA(50mL*3)萃取。合并有机层用饱和食盐水洗涤，无水硫酸钠干燥，过滤，减压浓缩。通过制备高效液相色谱纯化残余物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酰胺(2)(2.8mg)。MS:m/z＝783[M+H]+ To a solution of tert-butyl 4-((4-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenyl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (2-5) (170 mg, 0.29 mmol) in DCM (10 mL) was added TFA (5 mL) at room temperature. After stirring at room temperature for 4 h, the mixture was concentrated. The residue was dissolved in DMF (20 mL), and 5-(((1r,4r)-4-aminocyclohexyloxy)quinoline-8-carbonitrile (1-7) (310 mg, 1.16 mmol, 4.0 eq.), HATU (221 mg, 0.58 mmol, 2.0 eq.) and DIPEA (150 mg, 1.16 mmol, 4.0 eq.) were added. After stirring at room temperature for 4 h, H 2 O (50mL), and extracted with EA (50mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative high performance liquid chromatography to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenyl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (2) (2.8 mg). MS: m/z=783[M+H] +

实施例3化合物3的制备
Example 3 Preparation of Compound 3

第一步：化合物3-2的制备
Step 1: Preparation of compound 3-2

室温下，向2-溴-4，5-二氟苯甲酸(11.4g，48.3mmol，1.0eq.)的SOCl2(45mL)溶液中加入0.38mL DMF。在80℃下搅拌2小时后，冷至室温，浓缩至干燥，然后依次加入150mL DCM和DEA(17.6g,242mmol,5.0eq.)。室温下搅拌6小时后，加入45mL饱和NaHCO3水溶液和45mL饱和食盐水洗涤混合物。有机层用无水硫酸钠干燥，减压过滤浓缩，得到粗品2-溴-N，N-二乙基-4，5-二氟苯甲酰胺(3-2)(13.7g，粗品)，直接用于下一步。MS:m/z＝292[M+H]+ At room temperature, 0.38 mL of DMF was added to a solution of 2-bromo-4,5-difluorobenzoic acid (11.4 g, 48.3 mmol, 1.0 eq.) in SOCl 2 (45 mL). After stirring at 80°C for 2 hours, the mixture was cooled to room temperature and concentrated to dryness, and then 150 mL of DCM and DEA (17.6 g, 242 mmol, 5.0 eq.) were added in sequence. After stirring at room temperature for 6 hours, 45 mL of saturated aqueous NaHCO 3 solution and 45 mL of saturated brine were added to wash the mixture. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain a crude product of 2-bromo-N,N-diethyl-4,5-difluorobenzamide (3-2) (13.7 g, crude product), which was used directly in the next step. MS: m/z=292[M+H] +

第二步：化合物3-3的制备
Step 2: Preparation of compound 3-3

室温下，向2-溴-N，N-二乙基-4，5-二氟苯甲酰胺(3-2)(13.7g，粗品，1.0eq.)在1，4-二氧六环(120mL)和H2O(30mL)中的溶液中加入乙烯基三氟硼酸钾(7.5g，56mmol，1.2eq.)，Pd(dppf)Cl2(1.6g，2.3mmol，0.05eq.)和K2CO3(15.2g，116mmol，2.5eq.)。在95℃下搅拌16小时后，冷至室温，滤去固体，减压浓缩。然后将残余物加入到150mL EtOAc和45mL H2O中。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤减压浓缩，得到粗N，N-二乙基-4，5-二氟-2-乙烯基苯甲酰胺(3-3)(10.5g，粗品)，直接用于下一步。MS:m/z＝240[M+H]+ To a solution of 2-bromo-N,N-diethyl-4,5-difluorobenzamide (3-2) (13.7 g, crude product, 1.0 eq.) in 1,4-dioxane (120 mL) and H 2 O (30 mL) were added potassium vinyl trifluoroborate (7.5 g, 56 mmol, 1.2 eq.), Pd(dppf)Cl 2 (1.6 g, 2.3 mmol, 0.05 eq.) and K 2 CO 3 (15.2 g, 116 mmol, 2.5 eq.) at room temperature. After stirring at 95° C. for 16 hours, the mixture was cooled to room temperature, the solid was filtered off, and the mixture was concentrated under reduced pressure. The residue was then added to 150 mL of EtOAc and 45 mL of H 2 O. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude N,N-diethyl-4,5-difluoro-2-vinylbenzamide (3-3) (10.5 g, crude product), which was used directly in the next step. MS: m/z = 240 [M+H] +

第三步：化合物3-4的制备Step 3: Preparation of compound 3-4

室温下，向1,4-dioxane(100mL)和H2O(40mL)中依次加入N，N-二乙基-4，5-二氟-2-乙烯基苯甲酰胺(3-3)(10.5g，44mmol，1.0eq.)，K2OsO4·2H2O(0.83g，1.7mmol，0.04eq.)和NaIO4(18.2g，110mmol，2.5eq.)。室温下搅拌2小时后，将混合物过滤，减压浓缩。然后将残余物加入到150mL EtOAc和45mL H2O中。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物用硅胶柱色谱纯化PE/EA(50/1至1/1)，得到N，N-二乙基-4，5-二氟-2-甲酰基苯甲酰胺(3-4)(3.9g)。MS:m/z＝242[M+H]+ At room temperature, N,N-diethyl-4,5-difluoro-2-vinylbenzamide (3-3) (10.5 g, 44 mmol, 1.0 eq.), K 2 OsO4·2H 2 O (0.83 g, 1.7 mmol, 0.04 eq.) and NaIO4 (18.2 g, 110 mmol, 2.5 eq.) were added to 1,4-dioxane (100 mL) and H 2 O (40 mL) in sequence. After stirring at room temperature for 2 hours, the mixture was filtered and concentrated under reduced pressure. The residue was then added to 150 mL of EtOAc and 45 mL of H 2 O. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with PE/EA (50/1 to 1/1) to give N,N-diethyl-4,5-difluoro-2-formylbenzamide (3-4) (3.9 g). MS: m/z = 242 [M + H] +

第四步：化合物3-5的制备
Step 4: Preparation of compound 3-5

室温下，向N，N-二乙基-4，5-二氟-2-甲酰基苯甲酰胺(3-4)(3.9g，21.4mmol，1.0eq.)的AcOH(11mL)溶液中加入85％水合肼(1.1mL，27.8mmol，1.3eq.)。在145℃搅拌1h后，冷却至室温，过滤，得到粗品6，7-二氟酞嗪-1(2H)-酮(3-5)(2.3克，粗品)，直接用于下一步反应。MS:m/z＝183[M+H]+ At room temperature, 85% hydrazine hydrate (1.1 mL, 27.8 mmol, 1.3 eq.) was added to a solution of N,N-diethyl-4,5-difluoro-2-formylbenzamide (3-4) (3.9 g, 21.4 mmol, 1.0 eq.) in AcOH (11 mL). After stirring at 145°C for 1 h, the mixture was cooled to room temperature and filtered to obtain a crude product of 6,7-difluorophthalazin-1(2H)-one (3-5) (2.3 g, crude product), which was directly used in the next reaction. MS: m/z = 183 [M+H] +

第五步：化合物3-6的制备Step 5: Preparation of Compound 3-6

室温下，向6，7-二氟酞嗪-1(2H)-酮(3-5)(1.2g，6.6mmol，1.0eq.)的DMF(20mL)中加入NaH(60％ Wt,712mg,17.8mmol,2.7eq.)。在38℃下搅拌1小时后，将3-溴哌啶-2，6-二酮(1.9g，9.9mmol，1.5eq.)在DMF(4mL)的溶液滴加到混合物中。在38℃下搅拌3小时后，冷至室温，加入H2O(140mL)，并用EA(400mL)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤减压浓缩，得到粗品3-(6，7-二氟-1-氧代酞嗪-2(1H)-基)哌啶-2，6-二酮(3-6)(1.3g，粗品)，直接用于下一步反应。MS:m/z＝294[M+H]+ At room temperature, NaH (60% Wt, 712 mg, 17.8 mmol, 2.7 eq.) was added to a solution of 6,7-difluorophthalazin-1(2H)-one (3-5) (1.2 g, 6.6 mmol, 1.0 eq.) in DMF (20 mL). After stirring at 38°C for 1 hour, a solution of 3-bromopiperidine-2,6-dione (1.9 g, 9.9 mmol, 1.5 eq.) in DMF (4 mL) was added dropwise to the mixture. After stirring at 38°C for 3 hours, the mixture was cooled to room temperature, H 2 O (140 mL) was added, and the mixture was extracted with EA (400 mL). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain a crude product of 3-(6,7-difluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (3-6) (1.3 g, crude product), which was directly used in the next step. MS: m/z = 294 [M + H] +

第六步：化合物3-7的制备
Step 6: Preparation of compound 3-7

室温下，向3-(6，7-二氟-1-氧代酞嗪-2(1H)-基)哌啶-2，6-二酮(3-6)(1.3克粗品，4.4mmol，1.0eq.)的DMSO(19mL))溶液中中加入哌嗪-1-羧酸叔丁酯(1.65g，8.8mmol，2.0eq.)和DIPEA(2毫升)。在138℃下搅拌2小时后，冷至室温，向混合物中加入450mL EtOAc和130mL水。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物用硅胶柱纯化PE/EA(100/1至1/2)，得到4-(2-(2，6-二氧代哌啶基)-7-氟-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-羧酸叔丁酯(3-7)(0.8g)。MS:m/z＝460[M+H]+ To a solution of 3-(6,7-difluoro-1-oxophthalazin-2(1H)-yl)piperidine-2,6-dione (3-6) (1.3 g crude product, 4.4 mmol, 1.0 eq.) in DMSO (19 mL) at room temperature, piperazine-1-carboxylic acid tert-butyl ester (1.65 g, 8.8 mmol, 2.0 eq.) and DIPEA (2 ml) were added. After stirring at 138°C for 2 hours, the mixture was cooled to room temperature, and 450 mL of EtOAc and 130 mL of water were added to the mixture. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column with PE/EA (100/1 to 1/2) to give tert-butyl 4-(2-(2,6-dioxopiperidinyl)-7-fluoro-1-oxo-1,2-dihydrophthalazin-6-yl)piperazine-1-carboxylate (3-7) (0.8 g). MS: m/z=460 [M+H] +

第七步：化合物3-8的制备Step 7: Preparation of compound 3-8

室温下，向4-(2-(2，6-二氧代哌啶基)-7-氟-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-羧酸叔丁酯(3-7)(350mg，0.76mmol)的EA(2mL))溶液中加入盐酸(5mL,4M in EA)。室温下搅拌1小时后，浓缩得到3-(7-氟-1-氧代-6-(哌嗪-1-基)酞嗪-2(1H-基)哌啶-2，6-二酮盐酸盐(3-8)(0.30g)。MS:m/z＝360[M+H]+ At room temperature, hydrochloric acid (5 mL, 4 M in EA) was added to a solution of tert-butyl 4-(2-(2,6-dioxopiperidinyl)-7-fluoro-1-oxo-1,2-dihydrophthalazin-6-yl)piperazine-1-carboxylate (3-7) (350 mg, 0.76 mmol) in EA (2 mL). After stirring at room temperature for 1 hour, the mixture was concentrated to give 3-(7-fluoro-1-oxo-6-(piperazin-1-yl)phthalazin-2(1H-yl)piperidine-2,6-dione hydrochloride (3-8) (0.30 g). MS: m/z=360 [M+H] +

第八步：化合物3-9的制备
Step 8: Preparation of Compound 3-9

室温下，向3-(7-氟-1-氧代-6-(哌嗪-1-基)酞嗪-2(1H-基)哌啶-2，6-二酮(3-8)(160mg，0.4mmol，1.0eq.)在DCM/MeOH(6mL/4mL))的溶液中，中加入4-(4-甲酰基哌啶-1-基)苯甲酸叔丁酯(1-5)(110mg，0.38mmol，0.95eq.)和TEA(16滴)。在室温下搅拌3小时后，加入NaHB(OAc)3(270mg，1.08mmol，2.7eq.)。室温下搅拌2小时后，浓缩至干。向残余物中加入EA/H2O(20mL/20mL)，并用EA(20mL*3)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤浓缩，得到粗品叔丁基4-((4-(2-(2，6-二氧代哌啶-3-基)-7-氟-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸酯(3-9)(230mg，粗品)，用于下一步，无需纯化。MS:m/z＝633[M+H]+ To a solution of 3-(7-fluoro-1-oxo-6-(piperazin-1-yl)phthalazine-2(1H-yl)piperidine-2,6-dione (3-8) (160 mg, 0.4 mmol, 1.0 eq.) in DCM/MeOH (6 mL/4 mL) at room temperature, tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (1-5) (110 mg, 0.38 mmol, 0.95 eq.) and TEA (16 drops) were added. After stirring at room temperature for 3 hours, NaHB(OAc) 3 (270 mg, 1.08 mmol, 2.7 eq.) was added. After stirring at room temperature for 2 hours, the mixture was concentrated to dryness. EA/H 2 O (20 mL/20 mL) was added to the residue, and extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated to give crude tert-butyl 4-((4-(2-(2,6-dioxopiperidin-3-yl)-7-fluoro-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (3-9) (230 mg, crude product), which was used in the next step without purification. MS: m/z=633[M+H] +

第九步：化合物3的制备Step 9: Preparation of compound 3

室温下，向4-((4-(2-(2，6-二氧代哌啶-3-基)-7-氟-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(3-9)(110mg，0.17mmol，1.0eq.)的DCM(4mL)溶液中，加入TFA(2毫升)。室温下搅拌1h后，浓缩。将残余物溶解在DMF(5mL)中，并加入5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(56mg，0.19mmol，1.1eq.)，HATU(73mg，0.19毫摩尔，1.1eq.)和DIPEA(219毫克，1.7毫摩尔，10.0eq.)，室温下搅拌1h后，用H2O(20mL)稀释混合物并用取EA(20mL*3)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。通过制备型HPLC(HCOOH缓冲液)纯化残余物，得到N-((1r，4r)-((8-氰基喹啉-5-基)氧基)环己基) 4-((4-(2-(2-(2,6-二氧代哌啶-3-基)-7-氟-1-氧代-1,2-二氢邻苯二甲酸-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酰胺(3)(11mg)。MS:m/z＝826.3[M+H]+ To a solution of tert-butyl 4-((4-(2-(2,6-dioxopiperidin-3-yl)-7-fluoro-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzoate (3-9) (110 mg, 0.17 mmol, 1.0 eq.) in DCM (4 mL) at room temperature was added TFA (2 mL). The mixture was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DMF (5 mL), and 5-(((1r, 4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (56 mg, 0.19 mmol, 1.1 eq.), HATU (73 mg, 0.19 mmol, 1.1 eq.) and DIPEA (219 mg, 1.7 mmol, 10.0 eq.) were added. After stirring at room temperature for 1 h, the mixture was diluted with H 2 O (20 mL) and extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (HCOOH buffer) to give N-((1r, 4r)-((8-cyanoquinolin-5-yl)oxy)cyclohexyl) 4-((4-(2-(2-(2,6-dioxopiperidin-3-yl)-7-fluoro-1-oxo-1,2-dihydrophthalic acid-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (3) (11 mg). MS: m/z=826.3 [M+H] +

1H NMR(400MHz,dmso)δ11.02(s,1H),9.07(d,J＝4.2Hz,1H),8.64(dd,J＝8.6,0.9Hz,1H),8.36(s,1H),8.28(d,J＝8.5Hz,1H),7.98(d,J＝7.7Hz,1H),7.84(d,J＝13.3Hz,1H),7.74(d,J＝8.6Hz,2H),7.68(dd,J＝8.7,4.2Hz,1H),7.51(d,J＝8.1Hz,1H),7.35(d,J＝8.3Hz,1H),6.93(d,J＝8.5Hz,2H),5.77(dd,J＝11.6,7.0Hz,1H),4.79–4.66(m,1H),3.85(d,J＝13.5Hz,3H),3.26–3.22(m,4H),2.98–2.86(m,1H),2.77(t,J＝10.9Hz,2H),2.57(br,4H),2.30–2.19(m,4H),1.98–1.92(m,2H),1.86–1.76(m,3H),1.70–1.55(m,4H),1.28–1.20(m,5H). 1 H NMR(400MHz,dmso)δ11.02(s,1H),9.07(d,J＝4.2Hz,1H),8.64(dd,J＝8.6,0.9Hz,1H),8.36(s,1H),8.28( d,J＝8.5Hz,1H),7.98(d,J＝ 7.7Hz,1H),7.84(d,J＝13.3Hz,1H),7.74(d,J＝8.6Hz,2H),7.68(dd,J＝8.7,4.2Hz,1H),7.51(d,J＝ 8.1Hz,1H),7.35(d,J＝8.3Hz,1H),6 .93(d,J＝8.5Hz,2H),5.77(dd,J＝11.6,7.0Hz,1H),4.79–4.66(m,1H),3.85(d,J＝13.5Hz,3H),3.26– 3.22(m,4H),2.98–2.86(m,1H), 2.77(t,J＝10.9Hz,2H),2.57(br,4H),2.30–2.19(m,4H),1.98–1.92(m,2H),1.86–1.76(m,3H),1.70–1.55( m,4H),1.28–1.20(m,5H).

实施例4化合物4的制备
Example 4 Preparation of Compound 4

第一步：化合物4-2的制备
Step 1: Preparation of compound 4-2

在0℃，向(1R，5S，6r)-3-(叔丁氧羰基)-3-氮杂双环[3.1.0]己烷-6-羧酸(4-1)(2.0g，8.8mmol，1.0eq.)的四氢呋喃(20mL)溶液中滴加BH3·THF(1M THF，44mL，44mmol，5.0eq.)。在0℃下搅拌继续2小时，用甲醇(15.0mL)淬灭反应并浓缩。残余物用饱和碳酸钠(5.0mL)稀释，用乙酸乙酯(2×30mL)萃取。将合并的有机层用水洗涤，用无水硫酸钠干燥，过滤浓缩，得到叔丁基(1R，5S，6r)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-羧酸酯(4-2)(1.6g)。MS:m/z＝213[M+H]+ To a solution of (1R, 5S, 6r)-3-(tert-butyloxycarbonyl)-3-azabicyclo[3.1.0]hexane-6-carboxylic acid (4-1) (2.0 g, 8.8 mmol, 1.0 eq.) in tetrahydrofuran (20 mL) was added dropwise BH 3 ·THF (1M THF, 44 mL, 44 mmol, 5.0 eq.) at 0°C. Stirring was continued at 0°C for 2 hours, the reaction was quenched with methanol (15.0 mL) and concentrated. The residue was diluted with saturated sodium carbonate (5.0 mL) and extracted with ethyl acetate (2×30 mL). The combined organic layers were washed with water, dried over anhydrous sodium sulfate, filtered and concentrated to give tert-butyl (1R, 5S, 6r)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (4-2) (1.6 g). MS: m/z=213[M+H] +

第二步：化合物4-3的制备
Step 2: Preparation of compound 4-3

室温下，向叔丁基(1R，5S，6r)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-羧酸酯(4-2)(1.5g，7.5mmol，1.0eq.)的DCM(15mL)溶液中加入4M HCl in dioxane(8mL,2mmol)。室温下搅拌2小时后，减压浓缩，得到((1R，5S，6r)-3-氮杂双环[3.1.0]己烷-6-基)甲醇盐酸盐(1.5克)。To a solution of tert-butyl (1R, 5S, 6r)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (4-2) (1.5 g, 7.5 mmol, 1.0 eq.) in DCM (15 mL) was added 4 M HCl in dioxane (8 mL, 2 mmol) at room temperature. After stirring at room temperature for 2 hours, the mixture was concentrated under reduced pressure to give ((1R, 5S, 6r)-3-azabicyclo[3.1.0]hexane-6-yl)methanol hydrochloride (1.5 g).

向((1R，5S，6r)-3-氮杂双环[3.1.0]己烷-6-基)甲醇盐酸盐(1.5g，13.2mmol，1.0eq.)的DMF(15mL)溶液中加入4-氟苯甲酸叔丁酯(1.02g，14.5mmol，1.1eq)，然后加入K2CO3(7.2g，52.8mmol，4.0eq)。在140℃下搅拌4h后，冷至室温，将混合物分散到H2O(100mL)和EA(100mL)中，并用EA(3x 100mL)萃取。将有机层干燥并蒸发。残余物用硅胶柱纯化PE/EA(10：1至3：1)， 得到4-((1R，5S，6r)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)苯甲酸叔丁酯(4-3)(1.1g)。MS:m/z＝289[M+H]+ To a solution of ((1R, 5S, 6r)-3-azabicyclo[3.1.0]hexane-6-yl)methanol hydrochloride (1.5 g, 13.2 mmol, 1.0 eq.) in DMF (15 mL) was added tert-butyl 4-fluorobenzoate (1.02 g, 14.5 mmol, 1.1 eq) and then K 2 CO 3 (7.2 g, 52.8 mmol, 4.0 eq). After stirring at 140° C. for 4 h, it was cooled to room temperature, and the mixture was dispersed in H 2 O (100 mL) and EA (100 mL), and extracted with EA (3 x 100 mL). The organic layer was dried and evaporated. The residue was purified by silica gel column PE/EA (10:1 to 3:1), Obtained tert-butyl 4-((1R, 5S, 6r)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexane-3-yl)benzoate (4-3) (1.1 g). MS: m/z = 289 [M+H] +

第三步：化合物4-4的制备Step 3: Preparation of compound 4-4

室温下，向4-((1R，5S，6r)-6-(羟甲基)-3-氮杂双环[3.1.0]己-3-基)苯甲酸叔丁酯(4-3)(1.1克，3.8毫摩尔，1.0eq.))的DMSO(15毫升)溶液中，加入IBX(1.1克，4.18毫摩尔，1.1eq.)。室温下搅拌12h后，加入水(10mL)，白色沉淀生成。然后将混合物过滤并干燥滤饼，得到4-((1R，5S，6r)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)苯甲酸叔丁酯(4-4)(1.0g)。MS:m/z＝287[M+H]+ To a solution of tert-butyl 4-((1R, 5S, 6r)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hex-3-yl)benzoate (4-3) (1.1 g, 3.8 mmol, 1.0 eq.)) in DMSO (15 ml) at room temperature, IBX (1.1 g, 4.18 mmol, 1.1 eq.) was added. After stirring at room temperature for 12 h, water (10 mL) was added to form a white precipitate. The mixture was then filtered and the filter cake was dried to give tert-butyl 4-((1R, 5S, 6r)-6-formyl-3-azabicyclo[3.1.0]hex-3-yl)benzoate (4-4) (1.0 g). MS: m/z=287[M+H] +

第四步：化合物4-6的制备
Step 4: Preparation of Compound 4-6

室温下，向4-(1R，5S，6r)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)苯甲酸叔丁酯(4-4)(300mg，1.04mmol，1.0eq.)的DCM(3mL)溶液中加入3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮(4-5)(354mg，1.04毫摩尔，1.0eq.)和AcOH(6.0毫克，0.1毫摩尔，0.1eq.)。室温下搅拌4小时，然后加入NaBH(OAc)3(219mg，1.04mmol，1.0eq.)。室温下继续搅拌4小时，然后在减压下浓缩至干。向残余物加入H2O(20mL)并用EA(20mL*3)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并浓缩。残余物在DCM：EA(1：1，3mL)中洗涤，得到叔丁基4-((1R，5S，6S)-6-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢酞嗪-6-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)苯甲酸酯(4-6)(280毫克)，直接用于下一步反应。MS:m/z＝612[M+H]+ To a solution of tert-butyl 4-(1R,5S,6r)-6-formyl-3-azabicyclo[3.1.0]hexane-3-yl)benzoate (4-4) (300 mg, 1.04 mmol, 1.0 eq.) in DCM (3 mL) at room temperature, 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione (4-5) (354 mg, 1.04 mmol, 1.0 eq.) and AcOH (6.0 mg, 0.1 mmol, 0.1 eq.) were added. The mixture was stirred at room temperature for 4 hours, and then NaBH(OAc)3 (219 mg, 1.04 mmol, 1.0 eq.) was added. The mixture was stirred at room temperature for another 4 hours, and then concentrated to dryness under reduced pressure. H2O (20 mL) was added to the residue, and the mixture was extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated. The residue was washed in DCM:EA (1:1, 3 mL) to give tert-butyl 4-((1R,5S,6S)-6-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)-3-azabicyclo[3.1.0]hexane-3-yl)benzoate (4-6) (280 mg), which was used directly in the next step. MS: m/z=612[M+H] +

第五步：化合物4-7的制备
Step 5: Preparation of Compound 4-7

室温下，向4-((1R，5S，6S)-6-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)苯甲酸叔丁酯(4-6)(280毫克，0.5毫摩尔，1.0eq.)的DCM(4mL)溶液中加入4M HCl的二恶烷(2毫升)。室温下搅拌4小时后，减压浓缩，得到4-((1R，5S，6S)-6-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)苯甲酸(4-7)(270毫克)。To a solution of tert-butyl 4-((1R,5S,6S)-6-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)-3-azabicyclo[3.1.0]hexan-3-yl)benzoate (4-6) (280 mg, 0.5 mmol, 1.0 eq.) in DCM (4 mL) was added 4 M HCl in dioxane (2 mL) at room temperature. After stirring at room temperature for 4 hours, the mixture was concentrated under reduced pressure to give 4-((1R,5S,6S)-6-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)-3-azabicyclo[3.1.0]hexan-3-yl)benzoic acid (4-7) (270 mg).

第六步：化合物4的制备Step 6: Preparation of compound 4

室温下，向4-((1R，5S，6S)-6-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)苯甲酸(4-7)(120毫克，0.2毫摩尔，1.0eq.)， 5-(1r，4R)-4-氨基环己氧基)喹啉-8-甲腈(1-7)(53.4毫克，0.2毫摩尔，1.0eq.)的DMF(2毫升)溶液中加入DIEA(56毫克，0.4毫摩尔，2.0eq.)和HATU(91.2毫克、0.24毫摩尔、1.2eq.)。室温下搅拌2h后，加入H2O(20mL)并用EA(20mL*3)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。残余物通过反相色谱(HCOOH缓冲液)纯化，得到N-((1r，4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(((1R，5S，6S)-6-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)苯甲酰胺(4)(8.2毫克)。MS:m/z＝806[M+H]+ At room temperature, 4-((1R,5S,6S)-6-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)-3-azabicyclo[3.1.0]hexan-3-yl)benzoic acid (4-7) (120 mg, 0.2 mmol, 1.0 eq.) was added to the mixture. DIEA (56 mg, 0.4 mmol, 2.0 eq.) and HATU (91.2 mg, 0.24 mmol, 1.2 eq.) were added to a solution of 5-(1r, 4R)-4-aminocyclohexyloxy)quinoline-8-carbonitrile (1-7) (53.4 mg, 0.2 mmol, 1.0 eq.) in DMF (2 mL). After stirring at room temperature for 2 h, H 2 O (20 mL) was added and extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (HCOOH buffer) to give N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(((1R,5S,6S)-6-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)-3-azabicyclo[3.1.0]hexan-3-yl)benzamide (4) (8.2 mg). MS: m/z=806 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.00(s,1H),9.07(dd,J＝4.4,1.7Hz,1H),8.64(dd,J＝8.4,1.7Hz,1H),8.28(d,J＝8.4Hz,1H),8.26(s,1H),8.13–8.07(m,1H),7.92(d,J＝7.6Hz,1H),7.73(d,J＝8.5Hz,1H),7.68(dd,J＝8.5,4.3Hz,1H),7.56(d,J＝9.0Hz,1H),7.34(d,J＝9.0Hz,2H),6.55(d,J＝8.5Hz,2H),5.79–5.72(m,1H),4.72(br,1H),3.86(br,1H),3.63(d,J＝9.8Hz,2H),2.97–2.86(m,3H),2.64–2.50(m,6H),2.36–2.14(m,3H),2.12–2.05(m,1H),2.03–1.88(m,3H),1.80(s,1H),1.69–1.52(m,4H),1.47–1.38(m,1H),1.24–1.21(m,1H),1.02–0.93(m,1H). 1 H NMR (400MHz, DMSO-d 6 )δ11.00(s,1H),9.07(dd,J＝4.4,1.7Hz,1H),8.64(dd,J＝8.4,1.7Hz,1H),8.28(d,J＝8.4Hz,1H), 8.26(s,1H),8.13–8.07(m,1H),7. 92(d,J＝7.6Hz,1H),7.73(d,J＝8.5Hz,1H),7.68(dd,J＝8.5,4.3Hz,1H),7.56(d,J＝9.0Hz,1H), 7.34(d,J＝9.0Hz,2H),6.55(d,J＝8.5 Hz,2H),5.79–5.72(m,1H),4.72(br,1H),3.86(br,1H),3.63(d,J＝9.8Hz,2H),2.97–2.86(m,3H),2.64 –2.50(m,6H),2.36–2.14(m, 3H),2.12–2.05(m,1H),2.03–1.88(m,3H),1.80(s,1H),1.69–1.52(m,4H),1.47–1.38(m,1H),1.24–1.21( m,1H),1.02–0.93(m,1H).

实施例5化合物5的制备
Example 5 Preparation of Compound 5

室温下，向2-(2，6-二氧代哌啶-3-基)-5-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-1，3-二酮盐酸盐(5-4)(160毫克，0.406毫摩尔，1.0eq.)的DCM(3mL)溶液中加入N-((1r，4r)-4-((8-氰基喹啉-5-基)氧)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(1-7)(196毫克，0.406毫摩尔，1.0eq.)和TEA(62毫克，0.609毫摩尔，1.5eq.)。室温下搅拌4小时后，加入NaBH(OAc)3(86mg，0.406mmol，1.0eq.)。室温下继续搅拌4小时，减压浓缩至干。残余物加入H2O(20mL)并用EA(20mL*3)萃取。合并有机层用饱和食盐水洗涤，过滤，减压浓缩。残余物通过反相色谱(TFA缓冲液)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3S)-(((2，6-二氧代哌啶-3-基)-1,3-二氧代异吲哚-5-基)氧化基)甲基)吡咯烷-1-基)甲基哌啶-1-基)苯甲酰胺(5)(50mg,85％purity)。MS:m/z＝824[M+H]+ To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(((R)-pyrrolidin-3-yl)methoxy)isoindoline-1,3-dione hydrochloride (5-4) (160 mg, 0.406 mmol, 1.0 eq.) in DCM (3 mL) at room temperature were added N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (1-7) (196 mg, 0.406 mmol, 1.0 eq.) and TEA (62 mg, 0.609 mmol, 1.5 eq.). After stirring at room temperature for 4 hours, NaBH(OAc) 3 (86 mg, 0.406 mmol, 1.0 eq.) was added. Stirring was continued at room temperature for 4 hours and the mixture was concentrated to dryness under reduced pressure. The residue was added with H 2 O (20 mL) and extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3S)-(((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindol-5-yl)oxy)methyl)pyrrolidin-1-yl)methylpiperidin-1-yl)benzamide (5) (50 mg, 85% purity). MS: m/z=824[M+H] +

1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),9.07–9.01(m,1H),8.60(d,J＝1.5Hz,1H),8.26(dd,J＝8.4,1.4Hz,1H),7.98(d,J＝7.6Hz,1H),7.86(dd,J＝8.2,1.7Hz,1H),7.73(d,J＝8.4Hz,2H),7.67(ddd,J＝8.5,4.2,1.3Hz,1H),7.49–7.43(m,1H),7.38–7.29(m,2H),7.01–6.90(m,2H),5.11(dd,J＝12.6,5.4Hz,1H),4.71(br,1H),4.28–4.16(m,2H),3.86(d,J＝12.6Hz,3H),3.67(br,4H),3.12(d,J＝6.7Hz,3H),2.95(t,J＝11.3Hz,1H),2.87–2.73(m,3H),2.60(d,J＝3.7Hz,1H),2.48(d,J＝1.8Hz,2H),2.23(d,J＝10.9Hz,2H),1.94(br,3H),1.82(br,2H),1.66–1.52(m,4H),1.31–1.21(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ11.10(s,1H),9.07–9.01(m,1H),8.60(d,J＝1.5Hz,1H),8.26(dd,J＝8.4,1.4Hz,1H),7.98(d,J ＝7.6Hz,1H),7.86(dd,J＝8.2,1.7Hz,1H), 7.73(d,J＝8.4Hz,2H),7.67(ddd,J＝8.5,4.2,1.3Hz,1H),7.49–7.43(m,1H),7.38–7.29(m,2H),7.01–6.90( m,2H),5.11(dd,J＝12.6,5.4Hz, 1H),4.71(br,1H),4.28–4.16(m,2H),3.86(d,J＝12.6Hz,3H),3.67(br,4H),3.12(d,J＝6.7Hz,3H), 2.95(t,J＝11.3Hz,1H),2.87–2.73(m,3 H),2.60(d,J＝3.7Hz,1H),2.48(d,J＝1.8Hz,2H),2.23(d,J＝10.9Hz,2H),1.94(br,3H),1.82(br, 2H),1.66–1.52(m,4H),1.31–1.21(m,2H).

实施例6化合物6的制备
Example 6 Preparation of Compound 6

第一步：化合物6-1的制备
Step 1: Preparation of compound 6-1

将4-氟苯甲酸甲酯(5.00g，32.4mmol，1.0eq.)、1，4-二氧杂-8-氮杂螺[4.5]癸烷(6.97g，48.7mmol，1.5eq.)和K2CO3(8.97g，64.9mmol，2.0eq.)在DMSO(20mL)的混合物在120℃下搅拌16小时。冷至室温，加入EA(100毫升)。过滤混合物。滤液用水(100mL)洗涤。有机层浓缩。残余物用柱(PE/EA2：1)纯化，得到4-(1，4-二氧杂-8-氮杂螺[4.5]癸烷-8-基)苯甲酸甲酯(6-1)(4.90g)。A mixture of methyl 4-fluorobenzoate (5.00 g, 32.4 mmol, 1.0 eq.), 1,4-dioxa-8-azaspiro[4.5]decane (6.97 g, 48.7 mmol, 1.5 eq.) and K 2 CO 3 (8.97 g, 64.9 mmol, 2.0 eq.) in DMSO (20 mL) was stirred at 120° C. for 16 hours. After cooling to room temperature, EA (100 mL) was added. The mixture was filtered. The filtrate was washed with water (100 mL). The organic layer was concentrated. The residue was purified by column (PE/EA 2:1) to give methyl 4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)benzoate (6-1) (4.90 g).

第二步：化合物6-2的制备
Step 2: Preparation of compound 6-2

将4-(1，4-二氧杂-8-氮杂螺[4.5]癸烷-8-基)苯甲酸甲酯(6-1)(4.00g，14.4mmol，1.0eq)和2N HCl(7.2mL，14.4mmol，1.0eq.)在EtOH(30mL)中的溶液在75℃下搅拌16小时。冷至室温，加入EA(100mL)和水(100mL)。有机层浓缩。残余物经制备-TLC(纯EA)纯化，得到4-(4-氧代哌啶-1-基)苯甲酸甲酯(6-2)(1.33g)。A solution of methyl 4-(1,4-dioxa-8-azaspiro[4.5]decane-8-yl)benzoate (6-1) (4.00 g, 14.4 mmol, 1.0 eq) and 2N HCl (7.2 mL, 14.4 mmol, 1.0 eq.) in EtOH (30 mL) was stirred at 75 °C for 16 h. After cooling to room temperature, EA (100 mL) and water (100 mL) were added. The organic layer was concentrated. The residue was purified by preparative-TLC (pure EA) to give methyl 4-(4-oxopiperidin-1-yl)benzoate (6-2) (1.33 g).

第三步：化合物6-3的制备Step 3: Preparation of compound 6-3

室温下，向4-(4-氧代哌啶-1-基)苯甲酸甲酯(6-2)(550毫克，2.36毫摩尔，1.0eq.)的THF(5毫升)和水(5毫升)溶液中加入LiOH·H2O(300毫克，7.07毫摩尔，3.0eq.)。搅拌16小时后，加入EA(50毫升)和水(50毫升)。用HCl酸化水层至pH＝2，并用EA(50mL x 2)萃取。将合并的有机层浓缩得到4-(4-氧代哌啶-1-基)苯甲酸(6-3)(427mg)。To a solution of methyl 4-(4-oxopiperidin-1-yl)benzoate (6-2) (550 mg, 2.36 mmol, 1.0 eq.) in THF (5 mL) and water (5 mL) was added LiOH·H 2 O (300 mg, 7.07 mmol, 3.0 eq.) at room temperature. After stirring for 16 hours, EA (50 mL) and water (50 mL) were added. The aqueous layer was acidified to pH = 2 with HCl and extracted with EA (50 mL x 2). The combined organic layers were concentrated to give 4-(4-oxopiperidin-1-yl)benzoic acid (6-3) (427 mg).

第四步：化合物6-4的制备
Step 4: Preparation of compound 6-4

室温下，向4-(4-氧代哌啶-1-基)苯甲酸(6-3)(105mg，0.48mmol，1.0eq.)的DMF(1mL)溶液中加入DIEA(248mg，1.92mmol，4.0eq.)和HATU(200mg，0.53毫摩尔，1.1eq.)。10分钟后，加入5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(145mg，0.48mmol，1.0eq.)。搅拌16小时后， 加入EA(20毫升)和水(20毫升)。有机层浓缩。残余物通过制备PC-TLC(纯EA)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-氧代哌啶-1-基)苯甲酰胺(6-4)(57mg)。To a solution of 4-(4-oxopiperidin-1-yl)benzoic acid (6-3) (105 mg, 0.48 mmol, 1.0 eq.) in DMF (1 mL) at room temperature were added DIEA (248 mg, 1.92 mmol, 4.0 eq.) and HATU (200 mg, 0.53 mmol, 1.1 eq.). After 10 minutes, 5-(((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (145 mg, 0.48 mmol, 1.0 eq.) was added. After stirring for 16 hours, EA (20 mL) and water (20 mL) were added. The organic layer was concentrated. The residue was purified by preparative PC-TLC (pure EA) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-oxopiperidin-1-yl)benzamide (6-4) (57 mg).

第五步：化合物6的制备Step 5: Preparation of compound 6

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-氧代哌啶-1-基)苯甲酰胺(6-4)(47毫克，0.10毫摩尔，1.0eq.)和2-(2，6-二氧代哌啶-3-基)-5-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-1，3-二酮(5-4)(40毫克，0.10毫摩尔，1.0eq.)在DCM(1毫升)中的溶液中加入TEA(51毫克，0.50毫摩尔，5.0eq.)和MgSO4(120毫克，1.00毫摩尔，10eq.)。在25℃下搅拌1小时。加入NaBH3CN(16毫克，0.25毫摩尔，2.5eq.)。在25℃下搅拌继续16小时后，加入DCM(20毫升)和水(20毫升)。有机层浓缩。通过制备-TLC(DCM/MeOH 10：1)纯化残留物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-((3R)-3-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)哌啶-1-基)苯甲酰胺(6)(2mg)。MS:m/z＝810[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-oxopiperidin-1-yl)benzamide (6-4) (47 mg, 0.10 mmol, 1.0 eq.) and 2-(2,6-dioxopiperidin-3-yl)-5-(((R)-pyrrolidin-3-yl)methoxy)isoindoline-1,3-dione (5-4) (40 mg, 0.10 mmol, 1.0 eq.) in DCM (1 ml) at room temperature were added TEA (51 mg, 0.50 mmol, 5.0 eq.) and MgSO 4 (120 mg, 1.00 mmol, 10 eq.). The mixture was stirred at 25° C. for 1 hour. NaBH 3 CN (16 mg, 0.25 mmol, 2.5 eq.) was added. After stirring was continued for 16 hours at 25°C, DCM (20 mL) and water (20 mL) were added. The organic layer was concentrated. The residue was purified by preparative-TLC (DCM/MeOH 10:1) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-((3R)-3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)piperidin-1-yl)benzamide (6) (2 mg). MS: m/z=810 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),9.07(dd,J＝4.4,2.0Hz,1H),8.64(dd,J＝8.4,1.6Hz,1H),8.28(d,J＝8.4Hz,1H),8.01(d,J＝7.6Hz,1H),7.85(d,J＝8.4Hz,1H),7.76-7.67(m,3H),7.46(s,1H),7.38-7.34(m,2H),6.97(d,J＝8.4Hz,2H),5.14-5.09(m,1H),4.73(bs,1H),4.23-4.17(m,3H),3.93-3.84(m,3H),2.93-2.77(m,5H),2.67-2.57(m,2H),2.32-2.23(m,3H),2.06-1.90(m,6H),170-1.58(m,6H). 1 H NMR (400MHz, DMSO-d 6 )δ11.08(s,1H),9.07(dd,J＝4.4,2.0Hz,1H),8.64(dd,J＝8.4,1.6Hz,1H),8.28(d,J＝8.4Hz,1H) ,8.01(d,J＝7.6Hz,1H),7.85(d,J＝8.4Hz,1H),7.76-7.67(m,3H),7.46(s,1H),7.38-7.34(m,2H ),6.97(d,J＝8.4Hz,2H),5.14-5.09(m,1H),4.73(bs,1H),4.23-4.17(m,3H),3.93-3.84(m,3H) ,2.93-2.77(m,5H),2.67-2.57(m,2H),2.32-2.23(m,3H),2.06-1.90(m,6H),170-1.58(m,6H).

实施例7化合物7的制备
Example 7 Preparation of Compound 7

第一步：化合物7-1的制备Step 1: Preparation of compound 7-1

室温下，向叔丁基(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-羧酸酯(5-3)(100毫克，0.22毫摩尔，1.0eq.)的AcOH(15毫升)溶液中加入锌粉(257毫克，4.39毫摩尔，20eq.)。在80℃下搅拌16小时后，冷至室温，浓缩。残余物通过反相色谱(TFA缓冲液)纯化，得到叔丁基(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-3-羟基-1-氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-羧酸酯(7-1)(15mg)。MS:m/z＝385[M+H]+ To a solution of tert-butyl (3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (5-3) (100 mg, 0.22 mmol, 1.0 eq.) in AcOH (15 ml) at room temperature was added zinc powder (257 mg, 4.39 mmol, 20 eq.). After stirring at 80°C for 16 hours, the mixture was cooled to room temperature and concentrated. The residue was purified by reverse phase chromatography (TFA buffer) to give tert-butyl (3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-3-hydroxy-1-oxoisoindolin-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (7-1) (15 mg). MS: m/z=385[M+H] +

第二步：化合物7-2的制备
Step 2: Preparation of compound 7-2

室温下，向叔丁基(3R)-3-((((2-(2，6-二氧代哌啶-3-基)-3-羟基-1-氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-羧酸酯(7-1)(15mg，0.032毫摩尔，1.0eq.)和三乙基硅烷(15毫克，0.13毫摩尔，4.0eq.)在DCM(2毫升)中的溶液中加入TFA(1毫升)。室温下搅拌3小时后，浓缩得到3-(1-氧代-5-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-2-基)哌啶-2，6-二酮(7-2b)(15毫克，粗品)。MS: m/z＝343[M+H]+ To a solution of tert-butyl (3R)-3-((((2-(2,6-dioxopiperidin-3-yl)-3-hydroxy-1-oxoisoindolin-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (7-1) (15 mg, 0.032 mmol, 1.0 eq.) and triethylsilane (15 mg, 0.13 mmol, 4.0 eq.) in DCM (2 ml) was added TFA (1 ml) at room temperature. After stirring at room temperature for 3 hours, the mixture was concentrated to give 3-(1-oxo-5-(((R)-pyrrolidin-3-yl)methoxy)isoindolin-2-yl)piperidine-2,6-dione (7-2b) (15 mg, crude). MS: m/z＝343[M+H] +

第三步：化合物7的制备Step 3: Preparation of compound 7

室温下，向3-(1-氧代-5-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-2-基)哌啶-2，6-二酮(7-2)(15mg，0.043mmol，1.0eq.)的DMF(1mL)溶液中加入N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(20毫克，0.043毫摩尔，1.0eq.)和TEA(43毫克，0.43毫摩尔，10eq.)。将室温下搅拌4小时后，加入NaBH(OAc)3(86mg，0.406mmol，1.0eq.)。室温下搅拌4小时后，浓缩，加入H2O(20mL)并用EA(20mL*3)萃取。合并有机层用饱和食盐水洗涤，用Na2SO4干燥，过滤，减压浓缩。通过反相色谱(TFA缓冲液)纯化残余物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3S)-3-((2-(2,6-二氧代哌啶-3-基)-1-氧代异吲哚-5-基)甲基)吡咯烷-1-基)甲基(7)(16.3mg)。MS:m/z＝810[M+H]+ To a solution of 3-(1-oxo-5-(((R)-pyrrolidin-3-yl)methoxy)isoindolin-2-yl)piperidine-2,6-dione (7-2) (15 mg, 0.043 mmol, 1.0 eq.) in DMF (1 mL) at room temperature were added N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (20 mg, 0.043 mmol, 1.0 eq.) and TEA (43 mg, 0.43 mmol, 10 eq.). After stirring at room temperature for 4 hours, NaBH(OAc) 3 (86 mg, 0.406 mmol, 1.0 eq.) was added. After stirring at room temperature for 4 hours, the mixture was concentrated, H 2 O (20 mL) was added and extracted with EA (20 mL*3). The combined organic layers were washed with saturated brine, dried over Na 2 SO 4 , filtered, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3S)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindol-5-yl)methyl)pyrrolidin-1-yl)methyl (7) (16.3 mg). MS: m/z=810 [M+H] +

1H NMR(400MHz,DMSO-d6)δ10.98(s,1H),9.08(dd,J＝4.2,1.8Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8.29(d,J＝8.3Hz,1H),8.01(d,J＝7.6Hz,1H),7.76(d,J＝8.4Hz,2H),7.72–7.63(m,2H),7.36(d,J＝8.5Hz,1H),7.20(d,J＝2.5Hz,1H),7.09(dt,J＝8.5,2.5Hz,1H),7.00–6.93(m,2H),5.08(dd,J＝13.3,5.0Hz,1H),4.74(brs,1H),4.43–4.35(m,1H),4.33–4.24(m,1H),4.22–4.06(m,2H),3.94–3.78(m,4H),3.42–3.32(m,1H),3.23–3.12(m,3H),3.05–2.86(m,3H),2.85–2.76(m,3H),2.68–2.57(m,1H),2.44–2.31(m,1H),2.26(d,J＝11.1Hz,2H),2.19–2.08(m,1H),2.04–1.91(m,4H),1.88–1.78(m,2H),1.72–1.54(m,4H),1.35–1.23(m,2H) 1 H NMR (400 MHz, DMSO-d 6 )δ10.98(s,1H),9.08(dd,J＝4.2,1.8Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8 .29(d,J＝8.3Hz,1H),8.01(d,J＝7.6Hz,1H),7.76(d,J＝8.4Hz,2H),7.72–7.6 3(m,2H),7.36(d,J＝8.5Hz,1H),7.20(d,J＝2.5Hz,1H),7.09(dt,J＝8.5,2.5H z,1H),7.00–6.93(m,2H),5.08(dd,J＝13.3,5.0Hz,1H),4.74(brs,1H),4.43 –4.35(m,1H),4.33–4.24(m,1H),4.22–4.06(m,2H),3.94–3.78(m,4H),3.4 2–3.32(m,1H),3.23–3.12(m,3H),3.05–2.86(m,3H),2.85–2.76(m,3H),2.6 8–2.57(m,1H),2.44–2.31(m,1H),2.26(d,J＝11.1Hz,2H),2.19–2.08(m,1H) ,2.04–1.91(m,4H),1.88–1.78(m,2H),1.72–1.54(m,4H),1.35–1.23(m,2H)

实施例8化合物8的制备
Example 8 Preparation of Compound 8

第一步：化合物8-1的制备Step 1: Preparation of compound 8-1

在零度下，向叔丁基(R)-3-(羟甲基)吡咯烷-1-羧酸酯(3.0g，14.9mmol，1.0eq.)和TEA(3.01g，29.8mmol，2.0eq.)的DCM(30mL)溶液中加入MsCl(3.1g，26.8mmol，1.8eq.0)。室温搅拌4小时后，加入30mL饱和食盐水洗涤混合物。有机层用无水硫酸钠干燥，减压过滤浓缩，得到粗品叔丁基(R)-3-(((甲基磺酰基)氧基)甲基)吡咯烷-1-羧酸酯(8-1)(3.0克，粗品)，用于下一步，无需纯 化。MS:m/z＝280[M+H]+ At zero degrees, MsCl (3.1 g, 26.8 mmol, 1.8 eq.0) was added to a DCM (30 mL) solution of tert-butyl (R)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (3.0 g, 14.9 mmol, 1.0 eq.) and TEA (3.01 g, 29.8 mmol, 2.0 eq.). After stirring at room temperature for 4 hours, 30 mL of saturated brine was added to wash the mixture. The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to obtain crude tert-butyl (R)-3-(((methylsulfonyl)oxy)methyl)pyrrolidine-1-carboxylate (8-1) (3.0 g, crude), which was used in the next step without purification. MS: m/z = 280 [M + H] +

第二步：化合物8-2的制备Step 2: Preparation of compound 8-2

室温下，向(R)-3-(((甲基磺酰基)氧基)甲基)吡咯烷-1-羧酸叔丁酯(8-1)(3.0g，粗品，1.4eq.)和4-羟基苯甲酸甲酯(SM02)(1.61g，10.7mmol，1.0eq.)的DMF(20mL)溶液中加入K2CO3(2.9g，21.4mmol，2.0eq.)和NaI(0.8g，5.4mmol，0.5eq.)。在80℃下搅拌16小时后，冷至室温，加入用EtOAc(200mL)和饱和NaHCO3水溶液(60mL)。有机层用饱和NaHCO3水溶液和饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。硅胶柱色纯化PE/EA(100/1至2/1)，得到(R)-3-((4-(甲氧基羰基)苯氧基)甲基)吡咯烷-1-羧酸叔丁酯(8-2)(0.8g)。MS:m/z＝336[M+H]+ To a solution of (R)-tert-butyl 3-(((methylsulfonyl)oxy)methyl)pyrrolidine-1-carboxylate (8-1) (3.0 g, crude product, 1.4 eq.) and methyl 4-hydroxybenzoate (SM02) (1.61 g, 10.7 mmol, 1.0 eq.) in DMF (20 mL) at room temperature, K 2 CO 3 (2.9 g, 21.4 mmol, 2.0 eq.) and NaI (0.8 g, 5.4 mmol, 0.5 eq.) were added. After stirring at 80°C for 16 hours, the mixture was cooled to room temperature, and EtOAc (200 mL) and saturated aqueous NaHCO 3 solution (60 mL) were added. The organic layer was washed with saturated aqueous NaHCO 3 solution and saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. PE/EA (100/1 to 2/1) was purified by silica gel column chromatography to obtain (R)-tert-butyl 3-((4-(methoxycarbonyl)phenoxy)methyl)pyrrolidine-1-carboxylate (8-2) (0.8 g). MS: m/z=336 [M+H] +

第三步：化合物8-3的制备Step 3: Preparation of compound 8-3

室温下，向叔丁基(R)-3-((4-(甲氧基羰基)苯氧基)甲基)吡咯烷-1-羧酸叔丁酯(8-2)(1.8g，5.4mmol，1.0eq.)的DCM(14mL)溶液中加入TFA(3mL)。室温下搅拌1小时，浓缩得到(R)-4-(吡咯烷-3-基甲氧基)苯甲酸甲酯三氟乙酸盐(8-3)(2.0克)。MS:m/z＝236[M+H]+ At room temperature, TFA (3 mL) was added to a solution of tert-butyl (R)-3-((4-(methoxycarbonyl)phenoxy)methyl)pyrrolidine-1-carboxylate (8-2) (1.8 g, 5.4 mmol, 1.0 eq.) in DCM (14 mL). Stirred at room temperature for 1 hour, and concentrated to give (R)-4-(pyrrolidin-3-ylmethoxy)benzoic acid methyl ester trifluoroacetate (8-3) (2.0 g). MS: m/z = 236 [M+H] +

第四步：化合物8-5的制备Step 4: Preparation of compound 8-5

室温下，向(R)-4-(吡咯烷-3-基甲氧基)苯甲酸甲酯三氟乙酸盐(8-3)(2.0克，粗品，1.0eq.)和4-(4-(((((甲基磺酰基)氧基)甲基)哌啶-1-基)苯甲酸叔丁酯(2-3)(2.2克，6.0毫摩尔，1.1eq.)的DMF(8毫升)溶液中加入K2CO3(2.25克，16.4毫摩尔，3.0eq.)和NaI(0.56克，2.7毫摩尔，0.5eq.)。在80℃搅拌1小时后，冷至室温，加入用EtOAc(150mL)和H2O(45mL)。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物用硅胶柱纯化DCM/MeOH(200/1至20/1)得到无色油状4-(3R)-1-((1-(4-(叔丁氧羰基)环己-2，4-二烯-1-基)哌啶-4-基)甲基)吡咯烷-3-基)甲氧基)苯甲酸甲酯(8-5)(1.7克)。MS:m/z＝511[M+H]+ To a solution of (R)-4-(pyrrolidin-3-ylmethoxy)benzoic acid methyl ester trifluoroacetate (8-3) (2.0 g, crude, 1.0 eq.) and tert-butyl 4-(4-(((((methylsulfonyl)oxy)methyl)piperidin-1-yl)benzoate (2-3) (2.2 g, 6.0 mmol, 1.1 eq.) in DMF (8 mL) at room temperature were added K 2 CO 3 (2.25 g, 16.4 mmol, 3.0 eq.) and NaI (0.56 g, 2.7 mmol, 0.5 eq.). After stirring at 80° C. for 1 hour, the mixture was cooled to room temperature and a mixture of EtOAc (150 mL) and H 2 O (45 mL). The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column DCM/MeOH (200/1 to 20/1) to give colorless oily 4-(3R)-1-((1-(4-(tert-butyloxycarbonyl)cyclohexa-2,4-diene-1-yl)piperidin-4-yl)methyl)pyrrolidin-3-yl)methoxy)benzoic acid methyl ester (8-5) (1.7 g). MS: m/z=511[M+H] +

第五步：化合物8-6的制备Step 5: Preparation of compound 8-6

室温下，向4-(((3R)-1-((1-(4-(叔丁氧羰基)环己-2，4-二烯-1-基)哌啶-4-基)甲基)吡咯烷-3-基)甲氧基)苯甲酸甲酯(8-5)(2.0克，3.9毫摩尔，1.0eq.)的THF/H2O(18毫升/12毫升)溶液中加入LiOH·H2O(1.0克23.4毫摩尔,6.0eq.)。在45℃下搅拌40小时后，冷至室温，用1N HCl将混合物酸化至pH＝5，并用EA(50mL*3)萃取。将合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并减压浓缩，得到粗品4-(((3R)-1-((1-(4-(叔丁氧羰基)环己-2，4-二烯-1-基)哌啶-4-基)甲基)吡咯烷-3-基)甲氧基)苯甲酸(8-6)(1.8克，粗品)，用于下一步，无需纯化。MS:m/z＝497[M+H]+ To a solution of methyl 4-(((3R)-1-((1-(4-(tert-butyloxycarbonyl)cyclohexa-2,4-dien-1-yl)piperidin-4-yl)methyl)pyrrolidin-3-yl)methoxy)benzoate (8-5) (2.0 g, 3.9 mmol, 1.0 eq.) in THF/H2O (18 mL/12 mL) was added LiOH· H2O (1.0 g 23.4 mmol, 6.0 eq.) at room temperature. After stirring at 45°C for 40 hours, the mixture was cooled to room temperature, acidified to pH = 5 with 1N HCl, and extracted with EA (50 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give crude 4-(((3R)-1-((1-(4-(tert-butyloxycarbonyl)cyclohexa-2,4-dien-1-yl)piperidin-4-yl)methyl)pyrrolidin-3-yl)methoxy)benzoic acid (8-6) (1.8 g, crude), which was used in the next step without purification. MS: m/z=497[M+H] +

第六步：化合物8-7的制备Step 6: Preparation of compound 8-7

室温下，向4-(((3R)-1-((1-(4-((叔丁氧羰基)环己-2，4-二烯-1-基)哌啶-4-基)甲基)吡咯烷-3-基)甲氧基)苯甲酸(8-6)(1.8克，粗品，1.0eq.)和3-氨基哌啶-2，6-二酮(0.64克，3.8毫摩尔，1.05eq.)的DMF(12毫升)溶液中加入HATU(1.48克，3.8毫摩尔，1.05eq.)。和DIEA(2.4克，18.1毫摩尔，5.0eq.)。室温下搅拌3小时后，加H2O(100mL)，并用EA(100mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。残余物用Al2O3(pH＝9至10) 柱色谱法纯化，用(DCM：MeOH＝100/1至30/1)洗脱，得到4-(4-(3R)-3-((4-(2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸酯(8-7)(0.32g)。MS:m/z＝605[M+H]+ To a solution of 4-(((3R)-1-((1-(4-((tert-butyloxycarbonyl)cyclohexa-2,4-dien-1-yl)piperidin-4-yl)methyl)pyrrolidin-3-yl)methoxy)benzoic acid (8-6) (1.8 g, crude, 1.0 eq.) and 3-aminopiperidine-2,6-dione (0.64 g, 3.8 mmol, 1.05 eq.) in DMF (12 mL) at room temperature were added HATU (1.48 g, 3.8 mmol, 1.05 eq.) and DIEA (2.4 g, 18.1 mmol, 5.0 eq.). After stirring at room temperature for 3 hours, H 2 O (100 mL) was added, and the mixture was extracted with EA (100 mL*3). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by chromatography on a 4% HPLC -MS/MS system. Purification by column chromatography with (DCM:MeOH=100/1 to 30/1) gave 4-(4-(3R)-3-((4-(2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoate (8-7) (0.32 g). MS: m/z=605 [M+H] +

第七步：化合物8的制备Step 7: Preparation of compound 8

室温下，向4-(4-(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(8-7)(90毫克，0.15毫摩尔，1.0eq.)的DCM(4毫升)中的溶液中加入TFA(2毫升)。室温下搅拌1h，浓缩。将残留物溶解在DMF(4.5mL)中，所得混合物加入5-(((1r，4r)-4-氨基环己氧基)喹啉-8-甲腈(1-7)(50mg，0.17mmol，1.1eq.)，HATU(61mg，0.17mmol，1.1eq.)和DIPEA(193mg，1.5mmol，10.0eq.)。室温下搅拌2小时后，加H2O(20mL)，并用EA(20mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。通过制备型HPLC(HCOOH缓冲液)纯化残留物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3R)-3-((4-(2，6-二氧嘧啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基]哌啶-1-基)苯甲酰胺(8)(6mg)。MS:m/z＝798.4[M+H]+ To a solution of tert-butyl 4-(4-(3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoate (8-7) (90 mg, 0.15 mmol, 1.0 eq.) in DCM (4 mL) was added TFA (2 mL) at room temperature. The mixture was stirred at room temperature for 1 h and concentrated. The residue was dissolved in DMF (4.5 mL) and 5-(((1r,4r)-4-aminocyclohexyloxy)quinoline-8-carbonitrile (1-7) (50 mg, 0.17 mmol, 1.1 eq.), HATU (61 mg, 0.17 mmol, 1.1 eq.) and DIPEA (193 mg, 1.5 mmol, 10.0 eq.) were added to the resulting mixture. After stirring at room temperature for 2 h, H 2 O (20mL), and extracted with EA (20mL * 3). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by preparative HPLC (HCOOH buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3R)-3-((4-(2,6-dioxopyrimidine-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl]piperidin-1-yl)benzamide (8) (6 mg). MS: m/z = 798.4 [M + H] +

1H NMR(400MHz,DMSO-d6)δ10.80(s,1H),9.52–9.33(br,1H),7.99(d,J＝7.6Hz,1H),7.86(d,J＝8.8Hz,1H),7.74(d,J＝8.8Hz,2H),7.38(d,J＝2.4Hz,1H),7.14(dd,J＝8.8,2.4Hz,1H),6.95(d,J＝8.8Hz,2H),6.89(t,J＝9.3Hz,1H),6.54(dd,J＝15.1,2.0Hz,1H),6.45(dd,J＝8.5,2.4Hz,1H),4.58–4.47(m,1H),4.28(dd,J＝11.5,4.8Hz,1H),3.87(d,J＝12.6Hz,3H),3.58(d,J＝11.5Hz,3H),3.32–3.12(m,4H),3.13–2.99(m,4H),2.79(t,J＝12.1Hz,2H),2.76–2.64(m,1H),2.57(dt,J＝17.2,3.7Hz,1H),2.13–2.00(m,3H),1.90–1.79(m,3H),1.50(q,J＝11.5Hz,4H),1.33–1.23(m,3H). 1 H NMR (400MHz, DMSO-d 6 )δ10.80(s,1H),9.52–9.33(br,1H),7.99(d,J＝7.6Hz,1H),7.86(d,J＝8.8Hz,1H),7.74(d,J＝8.8 Hz,2H),7.38(d,J＝2.4Hz,1H),7.14(dd, J＝8.8,2.4Hz,1H),6.95(d,J＝8.8Hz,2H),6.89(t,J＝9.3Hz,1H),6.54(dd,J＝15.1,2.0Hz,1H),6.45( dd,J＝8.5,2.4Hz,1H),4.58–4.47(m, 1H),4.28(dd,J＝11.5,4.8Hz,1H),3.87(d,J＝12.6Hz,3H),3.58(d,J＝11.5Hz,3H),3.32–3.12(m,4H), 3.13–2.99(m,4H),2.79(t,J＝12.1H z,2H),2.76–2.64(m,1H),2.57(dt,J＝17.2,3.7Hz,1H),2.13–2.00(m,3H),1.90–1.79(m,3H),1.50(q, J＝11.5Hz,4H),1.33–1.23(m,3H).

实施例9化合物9的制备
Example 9 Preparation of Compound 9

第一步：化合物9-1的制备Step 1: Preparation of compound 9-1

室温下，向4-氟苯甲酸叔丁酯(5.0g，25.5mmol，1.0eq.)和哌啶-4-基甲醇(5.9g，51.0mmol，2.0eq.)的DMF(40mL)溶液中加入K2CO3(8.8g，51.0mmol，2.0eq.)。在118℃搅拌16小时后，冷至室温，加入H2O(60mL)和EtOAc(200mL)。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物用硅胶柱纯化PE/EA(50/1至1/2)，得到4-(4-(羟甲基)哌啶-1-基)苯甲酸叔丁酯(9-1)(5.9g)。MS:m/z＝292[M+H]+ To a solution of tert-butyl 4-fluorobenzoate (5.0 g, 25.5 mmol, 1.0 eq.) and piperidin-4-ylmethanol (5.9 g, 51.0 mmol, 2.0 eq.) in DMF (40 mL) was added K 2 CO 3 (8.8 g, 51.0 mmol, 2.0 eq.) at room temperature. After stirring at 118° C. for 16 hours, the mixture was cooled to room temperature, and H 2 O (60 mL) and EtOAc (200 mL) were added. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column PE/EA (50/1 to 1/2) to give tert-butyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate (9-1) (5.9 g). MS: m/z=292[M+H] +

第二步：化合物9-2的制备Step 2: Preparation of compound 9-2

室温下，向4-(4-(羟甲基)哌啶-1-基)苯甲酸叔丁酯(9-1)(4.5g，15.5mmol，1.0eq.)的二恶烷(64mL)溶液中加入HCl(84mL,4M in dioxane)。在75℃下搅拌16小时后，冷至室温，浓缩至干。将残留物在EA(40mL)中在室温下搅拌30分钟。将所得混合物过滤，滤饼用乙腈(40mL)洗涤并干燥，得到4-(4-(羟甲基)哌啶-1-基)苯甲酸(9-2)(3.6克)。MS:m/z＝236[M+H]+ At room temperature, HCl (84 mL, 4 M in dioxane) was added to a solution of tert-butyl 4-(4-(hydroxymethyl)piperidin-1-yl)benzoate (9-1) (4.5 g, 15.5 mmol, 1.0 eq.) in dioxane (64 mL). After stirring at 75 ° C for 16 hours, the mixture was cooled to room temperature and concentrated to dryness. The residue was stirred in EA (40 mL) at room temperature for 30 minutes. The resulting mixture was filtered, and the filter cake was washed with acetonitrile (40 mL) and dried to give 4-(4-(hydroxymethyl)piperidin-1-yl)benzoic acid (9-2) (3.6 g). MS: m/z = 236 [M + H] +

第三步：化合物9-4的制备Step 3: Preparation of compound 9-4

室温下，向4-(4-(羟甲基)哌啶-1-基)苯甲酸(9-2)(1.5g，6.4mmol，1.0eq.)和4-((((1r，4r)-4-氨基环己基)氧基)-2-氯苯腈(9-3)(1.6g，5.8mmol，0.9eq.)的DMF(36mL)溶液中加入HOAt(1.7g，12.8mmol，2.0eq.)，EDCI·盐酸(2.5克，12.8毫摩尔，2.0eq.)和DIPEA(4.2毫升，25.6毫摩尔，4.0eq.)。室温下搅拌3小时后，加H2O(80mL)并用EA(80mL*3)萃取。合并的有机层，用饱和食盐水洗涤，干燥，浓缩。将残留物在室温下在正庚烷/EA(50mL/10mL)中搅拌30分钟。将所得混合物过滤并干燥，得到N-((1r，4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-(羟甲基)哌啶-1-基)苯甲酰胺(9-4)(2.2克)。MS:m/z＝468[M+H]+ To a solution of 4-(4-(hydroxymethyl)piperidin-1-yl)benzoic acid (9-2) (1.5 g, 6.4 mmol, 1.0 eq.) and 4-((((1r,4r)-4-aminocyclohexyl)oxy)-2-chlorobenzonitrile (9-3) (1.6 g, 5.8 mmol, 0.9 eq.) in DMF (36 mL) were added HOAt (1.7 g, 12.8 mmol, 2.0 eq.), EDCI·hydrochloric acid (2.5 g, 12.8 mmol, 2.0 eq.) and DIPEA (4.2 mL, 25.6 mmol, 4.0 eq.) at room temperature. After stirring for 3 h at room temperature, H 2 O (80mL) and extracted with EA (80mL*3). The combined organic layer was washed with saturated brine, dried and concentrated. The residue was stirred in n-heptane/EA (50mL/10mL) at room temperature for 30 minutes. The resulting mixture was filtered and dried to give N-((1r, 4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-4-(4-(hydroxymethyl)piperidin-1-yl)benzamide (9-4) (2.2 g). MS: m/z=468[M+H] +

第四步：化合物9-5的制备Step 4: Preparation of compound 9-5

室温下，向N-((1r，4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-(羟甲基)哌啶-1-基)苯甲酰胺(9-4)(1.1g，2.4mmol，1.0eq.)的DCM(120mL)溶液中加入Dess-Martin试剂(1.2g，2.9mmol，1.2eq.)。在25℃下搅拌16小时后，加入饱和Na2S2 O3水溶液(60mL)中稀释，并用DCM(40mL*2)萃取。合并的有机层用饱和Na2S2O3水溶液(50mL)、饱和NaHCO3水溶液(50mL)和水(50mL)洗涤，用无水硫酸钠干燥，过滤减压浓缩，得到N-((1r，4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(9-5)(1.02g)。MS:m/z＝466[M+H]+ To a solution of N-((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-4-(4-(hydroxymethyl)piperidin-1-yl)benzamide (9-4) (1.1 g, 2.4 mmol, 1.0 eq.) in DCM (120 mL) was added Dess-Martin reagent (1.2 g, 2.9 mmol, 1.2 eq.) at room temperature. After stirring at 25°C for 16 hours, the mixture was diluted with saturated aqueous Na2S2O3 solution (60 mL) and extracted with DCM (40 mL * 2). The combined organic layer was washed with saturated aqueous Na2S2O3 solution (50 mL ), saturated aqueous NaHCO3 solution (50 mL) and water (50 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give N-((1r, 4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (9-5) (1.02 g). MS: m/z=466[M+H] +

第五步：化合物9-6的制备Step 5: Preparation of compound 9-6

室温下，向1，2-二氟-4-硝基苯(2.5g，15.7mmol，1.0eq.)和叔丁基哌嗪-1-羧酸酯(3.2g，17.3mmol，1.1eq.)的DMF(12mL)溶液中加入K2CO3(4.4g，31.4mmol，2.0eq.)。在60℃下搅拌3小时后，冷却至室温，将混合物倒入冰水(38mL)中并搅拌20分钟。将固体过滤并用水(15mL*2)洗涤，干燥得到4-(2-氟-4-硝基苯基)哌嗪-1-羧酸叔丁酯(9-6)(4.8克)。MS:m/z＝326[M+H]+ K 2 CO 3 (4.4 g, 31.4 mmol, 2.0 eq.) was added to a DMF (12 mL) solution of 1,2-difluoro-4-nitrobenzene (2.5 g, 15.7 mmol, 1.0 eq.) and tert-butylpiperazine-1-carboxylate (3.2 g, 17.3 mmol, 1.1 eq.) at room temperature. After stirring at 60°C for 3 hours, the mixture was cooled to room temperature and poured into ice water (38 mL) and stirred for 20 minutes. The solid was filtered and washed with water (15 mL*2) and dried to give tert-butyl 4-(2-fluoro-4-nitrophenyl)piperazine-1-carboxylate (9-6) (4.8 g). MS: m/z=326[M+H] +

第六步：化合物9-7的制备Step 6: Preparation of compound 9-7

室温下，向4-(2-氟-4-硝基苯基)哌嗪-1-羧酸叔丁酯(9-6)(2.0g，6.2mmol，1.0eq.)的EA(30mL)中加入Pd/C(5％ Wt,0.26g,0.69mmol,0.02eq.)。将反应混合物在H2气氛下于25℃搅拌16小时。 反应完成后，将混合物通过硅藻土垫过滤并用EA(10mL)洗涤。滤液真空浓缩，得4-(4-氨基-2-氟苯基)哌嗪-1-羧酸叔丁酯(9-7)(1.80克)。MS:m/z＝296[M+H]+ To tert-butyl 4-(2-fluoro-4-nitrophenyl)piperazine-1-carboxylate (9-6) (2.0 g, 6.2 mmol, 1.0 eq.) in EA (30 mL) was added Pd/C (5% Wt, 0.26 g, 0.69 mmol, 0.02 eq.) at room temperature. The reaction mixture was stirred at 25° C. under H 2 atmosphere for 16 hours. After the reaction was completed, the mixture was filtered through a celite pad and washed with EA (10 mL). The filtrate was concentrated in vacuo to give tert-butyl 4-(4-amino-2-fluorophenyl)piperazine-1-carboxylate (9-7) (1.80 g). MS: m/z = 296 [M+H] +

第七步：化合物9-8的制备Step 7: Preparation of compound 9-8

室温下，向4-(4-氨基-2-氟苯基)哌嗪-1-羧酸叔丁酯(9-7)(0.9g，3.1毫摩尔，1.0eq.)和3-溴哌啶-2，6-二酮(1.76g，9.3毫摩尔，3.0eq.)的DMF(6mL)溶液中加入Cs2CO3(2.5克，31毫摩尔，10.0eq.)。在85℃下搅拌24小时后，冷至室温，加入冰水(50mL)稀释混合物并用MTBE(50mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。残余物用硅胶柱色纯化PE/EA(50/1至1/2)，得到4-(4-((2，6-二氧代哌啶-3-基)氨基)-2-氟苯基)哌嗪-1-羧酸叔丁酯(9-8)(0.24g)。MS:m/z＝407[M+H]+ To a solution of tert-butyl 4-(4-amino-2-fluorophenyl)piperazine-1-carboxylate (9-7) (0.9 g, 3.1 mmol, 1.0 eq.) and 3-bromopiperidine-2,6-dione (1.76 g, 9.3 mmol, 3.0 eq.) in DMF (6 mL) at room temperature was added Cs 2 CO 3 (2.5 g, 31 mmol, 10.0 eq.). After stirring at 85° C. for 24 hours, the mixture was cooled to room temperature, ice water (50 mL) was added to dilute the mixture, and extracted with MTBE (50 mL*3). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with PE/EA (50/1 to 1/2) to give tert-butyl 4-(4-((2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazine-1-carboxylate (9-8) (0.24 g). MS: m/z=407 [M+H] +

第八步：化合物9-9的制备Step 8: Preparation of compound 9-9

室温下，向4-(4-(2，6-二氧代哌啶-3-基氨基)-2-氟苯基)哌嗪-1-羧酸叔丁酯(9-8)(120mg，0.30毫摩尔，1.0eq.)的dioxane(3mL)溶液中加入HCl(15mL,4M in dioxane)。在50℃搅拌16小时后，冷至室温，浓缩，得到3-((3-氟-4-(哌嗪-1-基)苯基)氨基)哌啶-2，6-二酮(9-9)(130毫克)。MS:m/z＝307[M+H]+ At room temperature, HCl (15 mL, 4 M in dioxane) was added to a solution of tert-butyl 4-(4-(2,6-dioxopiperidin-3-ylamino)-2-fluorophenyl)piperazine-1-carboxylate (9-8) (120 mg, 0.30 mmol, 1.0 eq.) in dioxane (3 mL). After stirring at 50°C for 16 hours, the mixture was cooled to room temperature and concentrated to give 3-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione (9-9) (130 mg). MS: m/z=307 [M+H] +

第九步：化合物9的制备Step 9: Preparation of compound 9

室温下，向3-((3-氟-4-(哌嗪-1-基)苯基)氨基)哌啶-2，6-二酮盐酸盐(9-9)(90毫克，0.26毫摩尔，1.0eq.)in DCM/MeOH(10mL/5mL)中加入N-((1r，4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(9-5)(108毫克，0.24毫摩尔，0.9eq.)和TEA(6滴)。在室温下搅拌16小时后，加入NaBH(OAc)3(132mg，0.65mmol，2.5eq.)。在室温下继续搅拌1小时，减压下浓缩至干。加入EA/H2O(20mL+20mL)，并用EA(20mL*2)提取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。通过反相色谱(TFA缓冲液)纯化残留物，得到N-((1r，4r)-4-(3-氯-4-氰基苯氧基)环己基)-4-(4-((4-(4-(2,6-二氧代哌啶-3-基)氨基)-2-氟苯基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酰胺(9)(75mg)。MS:m/z＝756.3[M+H]+ N-((1r,4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (9-5) (108 mg, 0.24 mmol, 0.9 eq.) and TEA (6 drops) were added to 3-((3-fluoro-4-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione hydrochloride (9-9) (90 mg, 0.26 mmol, 1.0 eq.) in DCM/MeOH (10 mL/5 mL) at room temperature. After stirring at room temperature for 16 hours, NaBH(OAc) 3 (132 mg, 0.65 mmol, 2.5 eq.) was added. Stirring was continued at room temperature for 1 hour, and the mixture was concentrated to dryness under reduced pressure. EA/H 2 O (20 mL+20 mL) was added, and the mixture was extracted with EA (20 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-(3-chloro-4-cyanophenoxy)cyclohexyl)-4-(4-((4-(4-(2,6-dioxopiperidin-3-yl)amino)-2-fluorophenyl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (9) (75 mg). MS: m/z=756.3[M+H] +

1H NMR(400MHz,DMSO-d6)δ10.80(s,1H),9.52–9.33(br,1H),7.99(d,J＝7.6Hz,1H),7.86(d,J＝8.8Hz,1H),7.74(d,J＝8.8Hz,2H),7.38(d,J＝2.4Hz,1H),7.14(dd,J＝8.8,2.4Hz,1H),6.95(d,J＝8.8Hz,2H),6.89(t,J＝9.3Hz,1H),6.54(dd,J＝15.1,2.0Hz,1H),6.45(dd,J＝8.5,2.4Hz,1H),4.58–4.47(m,1H),4.28(dd,J＝11.5,4.8Hz,1H),3.87(d,J＝12.6Hz,3H),3.58(d,J＝11.5Hz,3H),3.32–3.12(m,4H),3.13–2.99(m,4H),2.79(t,J＝12.1Hz,2H),2.76–2.64(m,1H),2.57(dt,J＝17.2,3.7Hz,1H),2.13–2.00(m,3H),1.90–1.79(m,3H),1.50(q,J＝11.5Hz,4H),1.33–1.23(m,3H). 1 H NMR (400MHz, DMSO-d 6 )δ10.80(s,1H),9.52–9.33(br,1H),7.99(d,J＝7.6Hz,1H),7.86(d,J＝8.8Hz,1H),7.74(d,J＝8.8 Hz,2H),7.38(d,J＝2.4Hz,1H),7.14(dd, J＝8.8,2.4Hz,1H),6.95(d,J＝8.8Hz,2H),6.89(t,J＝9.3Hz,1H),6.54(dd,J＝15.1,2.0Hz,1H),6.45( dd,J＝8.5,2.4Hz,1H),4.58–4.47(m, 1H),4.28(dd,J＝11.5,4.8Hz,1H),3.87(d,J＝12.6Hz,3H),3.58(d,J＝11.5Hz,3H),3.32–3.12(m,4H), 3.13–2.99(m,4H),2.79(t,J＝12.1H z,2H),2.76–2.64(m,1H),2.57(dt,J＝17.2,3.7Hz,1H),2.13–2.00(m,3H),1.90–1.79(m,3H),1.50(q, J＝11.5Hz,4H),1.33–1.23(m,3H).

实施例10化合物10的制备
Example 10 Preparation of Compound 10

第一步：化合物10-1的制备Step 1: Preparation of compound 10-1

室温下，向5-溴吲哚啉溶液(120毫克，0.6毫摩尔，1.0eq.)的DCM(2mL)中加入DIEA(234毫克，1.8毫摩尔，3.0eq.)和CbzCl(124毫克，0.7毫摩尔，1.2eq.)。室温搅拌4小时后，浓缩。残余物在用己烷/乙酸乙酯(8：1)洗脱的硅胶上快速色谱纯化，得到5-溴吲哚啉-1-羧酸苄酯(10-1)(195mg)。MS:m/z＝332[M+H]+ To a solution of 5-bromoindoline (120 mg, 0.6 mmol, 1.0 eq.) in DCM (2 mL) at room temperature were added DIEA (234 mg, 1.8 mmol, 3.0 eq.) and CbzCl (124 mg, 0.7 mmol, 1.2 eq.). After stirring at room temperature for 4 hours, the mixture was concentrated. The residue was purified by flash chromatography on silica gel eluted with hexane/ethyl acetate (8:1) to give 5-bromoindoline-1-carboxylic acid benzyl ester (10-1) (195 mg). MS: m/z = 332 [M+H] +

第二步：化合物10-2的制备Step 2: Preparation of compound 10-2

室温下，向5-溴吲哚啉-1-羧酸苄酯(10-1)(195毫克，0.61毫摩尔，1.0eq.)的甲苯溶液中加入哌嗪-1-羧酸叔丁酯(136.4毫克，0.73毫摩尔，1.2eq.)，NaOtBu(82毫克，0.85毫摩尔，1.4eq.)，XantPhos(28毫克，0.05毫摩尔，0.08eq.)和Pd2(dba)3(22毫克，0.02毫摩尔，0.04eq.)。将反应容器抽真空并回填氮气(三次)。在90℃下搅拌4小时后，冷却至室温，加入H2O(20mL)并EA(20mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。用己烷/乙酸乙酯(8：1)洗脱的硅胶上通过快速色谱纯化残余物，得到5-(4-(叔丁氧羰基)哌嗪-1-基)吲哚啉-1-羧酸苄酯(100mg)。MS:m/z＝438[M+H]+ To a toluene solution of 5-bromoindoline-1-carboxylic acid benzyl ester (10-1) (195 mg, 0.61 mmol, 1.0 eq.) was added tert-butyl piperazine-1-carboxylate (136.4 mg, 0.73 mmol, 1.2 eq.), NaOtBu (82 mg, 0.85 mmol, 1.4 eq.), XantPhos (28 mg, 0.05 mmol, 0.08 eq.) and Pd 2 (dba) 3 (22 mg, 0.02 mmol, 0.04 eq.) at room temperature. The reaction vessel was evacuated and backfilled with nitrogen (three times). After stirring at 90°C for 4 hours, the mixture was cooled to room temperature, H 2 O (20 mL) was added and extracted with EA (20 mL*3). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by flash chromatography on silica gel eluting with hexane/ethyl acetate (8:1) to give benzyl 5-(4-(tert-butoxycarbonyl)piperazin-1-yl)indoline-1-carboxylate (100 mg). MS: m/z = 438 [M+H] +

向5-(4-(叔丁氧羰基)哌嗪-1-基)吲哚啉-1-羧酸酯(100mg，0.3mmol，1.0eq.)in MeOH(2mL)中加入Pd/C(10.0mg，0.1eq.)。在H2气氛下搅拌6小时后，过滤混合物并用MeOH(5mL)洗涤。滤液浓缩得到白色固体的4-(吲哚啉-5-基)哌嗪-1-羧酸叔丁酯(10-2)(60毫克)。MS:m/z＝304[M+H]+ Pd/C (10.0 mg, 0.1 eq.) was added to 5-(4-(tert-butyloxycarbonyl)piperazine-1-yl)indoline-1-carboxylate (100 mg, 0.3 mmol, 1.0 eq.) in MeOH (2 mL). After stirring for 6 hours under H2 atmosphere, the mixture was filtered and washed with MeOH (5 mL). The filtrate was concentrated to give tert-butyl 4-(indoline-5-yl)piperazine-1-carboxylate (10-2) (60 mg) as a white solid. MS: m/z = 304 [M+H] +

第三步：化合物10-3的制备Step 3: Preparation of compound 10-3

室温下，向4-(吲哚啉-5-基)哌嗪-1-羧酸叔丁酯(10-2)(60.0mg，0.19mmol，1.0eq.)的MeCN(1mL)溶液中，加入3-溴哌啶-2，6-二酮(75mg，0.39mmol，2.0eq.)和NaHCO3(49mg，0.59毫摩尔，3eq.)。在95℃下搅拌2小时后，通过反相色谱(TFA缓冲液)纯化反应混合物，得到4-(1-(2，6-二氧代哌啶-3-基)吲哚啉-5-基)哌嗪-1-羧酸叔丁酯(10-3)(70mg)。To a solution of tert-butyl 4-(indolyl-5-yl)piperazine-1-carboxylate (10-2) (60.0 mg, 0.19 mmol, 1.0 eq.) in MeCN (1 mL) at room temperature, 3-bromopiperidine-2,6-dione (75 mg, 0.39 mmol, 2.0 eq.) and NaHCO 3 (49 mg, 0.59 mmol, 3 eq.) were added. After stirring at 95° C. for 2 hours, the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give tert-butyl 4-(1-(2,6-dioxopiperidin-3-yl)indolyl-5-yl)piperazine-1-carboxylate (10-3) (70 mg).

MS:m/z＝415[M+H]+ MS: m/z = 415 [M + H] +

第四步：化合物10-4的制备Step 4: Preparation of compound 10-4

室温下，向4-(1-(2，6-二氧代哌啶-3-基)吲哚啉-5-基)哌嗪-1-羧酸叔丁酯(10-3)(70mg，0.2mmol，1.0eq.)的DCM(3mL)溶液中加入4M HCl的EA(3mL)。室温下搅拌2小时后，减压浓缩，得到3-(5-(5-(哌嗪-1-基)吲哚啉-1-基)哌啶-2，6-二酮盐酸盐(10-4)(50毫克)。MS:m/z＝314[M+H]+ To a solution of tert-butyl 4-(1-(2,6-dioxopiperidin-3-yl)indolin-5-yl)piperazine-1-carboxylate (10-3) (70 mg, 0.2 mmol, 1.0 eq.) in DCM (3 mL) was added 4M HCl in EA (3 mL) at room temperature. After stirring at room temperature for 2 hours, the mixture was concentrated under reduced pressure to give 3-(5-(5-(piperazin-1-yl)indolin-1-yl)piperidine-2,6-dione hydrochloride (10-4) (50 mg). MS: m/z=314[M+H] +

第五步：化合物10的制备Step 5: Preparation of compound 10

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(52mg，0.1mmol，1eq.)的DCM(1mL)溶液中加入3-(5-((哌嗪-1-基)吲哚啉-1-基) 哌啶-2，6-二酮(10-4)(34毫克，0.1毫摩尔，1.0eq.)，TEA(一滴)和MgSO4(129毫克，1.0毫摩尔，10.0eq.)。室温下搅拌30分钟后，加入NaHB(OAc)3(68mg，0.3mmol，3.0eq.)。在室温下继续搅拌1小时后，通过反相色谱(TFA缓冲液)纯化反应混合物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(4-(1-(2，6-二氧代哌啶-3-基)吲哚啉-5-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酰胺(10)(9.5mg)。MS:m/z＝782[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (52 mg, 0.1 mmol, 1 eq.) in DCM (1 mL) was added 3-(5-((piperazin-1-yl)indolin-1-yl)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (52 mg, 0.1 mmol, 1 eq.) at room temperature. Piperidine-2,6-dione (10-4) (34 mg, 0.1 mmol, 1.0 eq.), TEA (one drop) and MgSO 4 (129 mg, 1.0 mmol, 10.0 eq.). After stirring at room temperature for 30 minutes, NaHB(OAc) 3 (68 mg, 0.3 mmol, 3.0 eq.) was added. After stirring at room temperature for 1 hour, the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(4-(1-(2,6-dioxopiperidin-3-yl)indolin-5-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (10) (9.5 mg). MS: m/z=782[M+H] +

1H NMR(400MHz,DMSO-d6)δ10.74(s,1H),9.05(dd,J＝4.0,1.8Hz,1H),8.62(dd,J＝8.6,1.4Hz,1H),8.27(d,J＝8.2Hz,1H),7.97(d,J＝7.6Hz,1H),7.72(s,1H),7.67(dd,J＝8.4,4.2Hz,1H),7.34(d,J＝8.4Hz,1H),6.91(d,J＝8.6Hz,2H),6.73(s,1H),6.54(d,J＝7.4Hz,1H),6.35(d,J＝8.4Hz,1H),4.74–4.68(m,1H),4.50(dd,J＝12.8,4.8Hz,1H),3.94–3.75(m,4H),2.97–2.89(m,3H),2.87–2.83(m,1H),2.80–2.69(m,3H),2.25–2.13(m,4H),2.00–1.88(m,4H),1.80–1.74(m,2H),1.66–1.50(m,5H),1.24–1.16(m,10H). 1 H NMR (400MHz, DMSO-d 6 )δ10.74(s,1H),9.05(dd,J＝4.0,1.8Hz,1H),8.62(dd,J＝8.6,1.4Hz,1H),8.27(d,J＝8.2Hz,1H), 7.97(d,J＝7.6Hz,1H),7. 72(s,1H),7.67(dd,J＝8.4,4.2Hz,1H),7.34(d,J＝8.4Hz,1H),6.91(d,J＝8.6Hz,2H),6.73(s,1H ),6.54(d,J＝7.4Hz,1H),6 .35(d,J＝8.4Hz,1H),4.74–4.68(m,1H),4.50(dd,J＝12.8,4.8Hz,1H),3.94–3.75(m,4H),2.97–2.89(m ,3H),2.87–2.83(m ,1H),2.80–2.69(m,3H),2.25–2.13(m,4H),2.00–1.88(m,4H),1.80–1.74(m,2H),1.66–1.50(m,5H),1.24 –1.16(m,10H).

实施例11化合物11的制备
Example 11 Preparation of Compound 11

第一步：化合物11-2的制备Step 1: Preparation of compound 11-2

室温下，向4-溴-2-氟苯腈(11-1)(15.0克，75.0毫摩尔，1.0eq.)的DMA(75毫升)溶液中加入MeNHNH2·H2SO4(13.0克，90.0毫摩尔，1.2eq.)和DIEA(33.9克，262.5毫摩尔，3.5eq.)。在150℃搅拌24小时后，冷至室温，加入EA(750mL)和水(750mL)。用EA(500mL)萃取。合并有机层真空浓缩。残余物用PE/EA(1：1)洗脱硅胶色谱纯化，得到6-溴-1-甲基-1H-吲唑-3-胺(11-2)(13.4g)。To a solution of 4-bromo-2-fluorobenzonitrile (11-1) (15.0 g, 75.0 mmol, 1.0 eq.) in DMA (75 mL) at room temperature were added MeNHNH 2 ·H 2 SO 4 (13.0 g, 90.0 mmol, 1.2 eq.) and DIEA (33.9 g, 262.5 mmol, 3.5 eq.). After stirring at 150°C for 24 hours, the mixture was cooled to room temperature, and EA (750 mL) and water (750 mL) were added. The mixture was extracted with EA (500 mL). The combined organic layers were concentrated in vacuo. The residue was purified by silica gel chromatography eluting with PE/EA (1:1) to give 6-bromo-1-methyl-1H-indazol-3-amine (11-2) (13.4 g).

第二步：化合物11-3的制备Step 2: Preparation of compound 11-3

室温下，将乳酸(420mg，85％，3.97mmol，1.3eq)和DBU(604mg，3.97mmol，1.3eq.)的混合物在25℃下搅拌16h，然后加入6-溴-1-甲基-1H-吲唑-3-胺(11-2)(670mg，2.96mmol，1.0eq.)和丙烯酸甲酯(1.84g，21.4mmol，7.0eq.)。在85℃搅拌16小时后，冷至室温，加入EA(20mL)和水(20mL)之间。浓缩有机层，残余物在用PE/EA(1：1)洗脱的硅胶上色谱纯化，得到3-((6-溴-1-甲基-1H-吲唑-3-氨基)丙酸甲酯(11-3)(650mg)。At room temperature, a mixture of lactic acid (420 mg, 85%, 3.97 mmol, 1.3 eq.) and DBU (604 mg, 3.97 mmol, 1.3 eq.) was stirred at 25°C for 16 h, and then 6-bromo-1-methyl-1H-indazole-3-amine (11-2) (670 mg, 2.96 mmol, 1.0 eq.) and methyl acrylate (1.84 g, 21.4 mmol, 7.0 eq.) were added. After stirring at 85°C for 16 hours, the mixture was cooled to room temperature and EA (20 mL) and water (20 mL) were added. The organic layer was concentrated and the residue was purified by chromatography on silica gel eluted with PE/EA (1:1) to give methyl 3-((6-bromo-1-methyl-1H-indazole-3-amino)propanoate (11-3) (650 mg).

第三步：化合物11-4的制备 Step 3: Preparation of compound 11-4

室温下，向3-((6-溴-1-甲基-1H-吲唑-3-氨基)丙酸酯(11-3)(650mg，2.08mmol，1.0eq.)的AcOH(6.5mL)中加入KCNO(338mg，4.16mmol，2.0eq.)。在25℃下搅拌16后，加入EA(50mL)和水(50mL)。浓缩有机层，残余物在用硅胶柱纯化EA/MeOH(10：1)，得到无色油状3-(1-(6-溴-1-甲基-1H-吲唑-3-基)脲基)丙酸甲酯(11-4)(518mg)。To a solution of 3-((6-bromo-1-methyl-1H-indazole-3-amino)propanoate (11-3) (650 mg, 2.08 mmol, 1.0 eq.) in AcOH (6.5 mL) was added KCNO (338 mg, 4.16 mmol, 2.0 eq.) at room temperature. After stirring at 25°C for 16, EA (50 mL) and water (50 mL) were added. The organic layer was concentrated and the residue was purified by silica gel column with EA/MeOH (10:1) to give methyl 3-(1-(6-bromo-1-methyl-1H-indazole-3-yl)ureido)propanoate (11-4) (518 mg) as a colorless oil.

第四步：化合物11-5的制备Step 4: Preparation of compound 11-5

室温下，向3-(1-(6-溴-1-甲基-1H-吲唑-3-基)脲基)丙酸酯(11-4)(485mg，1.37mmol，1.0eq.)的MeCN(5mL)溶液中加入Triton-B(400mg，40％，0.96mmol，0.7eq.)。在25℃下搅拌16h后，加入EA(50mL)和水(50mL)。有机层浓缩得到1-(6-溴-1-甲基-1H-吲唑-3-基)二氢嘧啶-2，4(1H，3H)-二酮(11-5)(364mg)。Triton-B (400 mg, 40%, 0.96 mmol, 0.7 eq.) was added to a solution of 3-(1-(6-bromo-1-methyl-1H-indazol-3-yl)ureido)propanoate (11-4) (485 mg, 1.37 mmol, 1.0 eq.) in MeCN (5 mL) at room temperature. After stirring at 25°C for 16 h, EA (50 mL) and water (50 mL) were added. The organic layer was concentrated to give 1-(6-bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (11-5) (364 mg).

第五步：化合物11-6的制备Step 5: Preparation of compound 11-6

室温下，向1-(6-溴-1-甲基-1H-吲唑-3-基)二氢嘧啶-2，4(1H，3H)-二酮(11-5)(1.00g，3.09mmol，1.0eq.)甲苯溶液中加入哌嗪-1-羧酸叔丁酯(690mg，3.71mmol，1.2eq.)、Pd2(dba)3(140mg、0.15mmol、0.05eq.)、XantPhos(180mg、0.31mmol，0.1eq.)和t-BuONa(890mg、9.28mmol、3.0eq.)。在85℃下搅拌4小时后，将反应物冷却至室温，用NH4Cl水溶液(20％，30mL)淬灭并用EA(30mL)萃取。有机层浓缩。用制备的TLC(纯EA)纯化残余物，得到哌嗪-1-羧酸叔丁酯，4-(3-(2，4-二氧代四氢嘧啶-1(2H)-基)-1-甲基-1H-吲唑-6-基)哌嗪-1-羧酸叔丁酯(11-6)(440mg)。MS:m/z＝429[M+H]+ To a toluene solution of 1-(6-bromo-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (11-5) (1.00 g, 3.09 mmol, 1.0 eq.) were added tert-butyl piperazine-1-carboxylate (690 mg, 3.71 mmol, 1.2 eq.), Pd 2 (dba) 3 (140 mg, 0.15 mmol, 0.05 eq.), XantPhos (180 mg, 0.31 mmol, 0.1 eq.) and t-BuONa (890 mg, 9.28 mmol, 3.0 eq.) at room temperature. After stirring at 85°C for 4 hours, the reaction was cooled to room temperature, quenched with aqueous NH 4 Cl solution (20%, 30 mL) and extracted with EA (30 mL). The organic layer was concentrated. The residue was purified by preparative TLC (pure EA) to give tert-butyl piperazine-1-carboxylate, 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate (11-6) (440 mg). MS: m/z=429 [M+H] +

第六步：化合物11-7的制备Step 6: Preparation of compound 11-7

室温下，向4-(3-(2，4-二氧代四氢嘧啶-1(2H)-基)-1-甲基-1H-吲唑-6-基)哌嗪-1-羧酸叔丁酯(10-6)(170mg，0.397mmol，1.0eq.)的EA(5mL)溶液中加入4M HCl in dioxane(2.5mL)。继续搅拌16小时，LCMS显示反应完成。将EA(20mL)加入混合物中，并在过滤前搅拌5分钟。滤饼干燥得到1-(1-甲基-6-(哌嗪-1-基)-1H-吲唑-3-基)二氢嘧啶-2，4(1H，3H)-二酮(11-7)(100mg)。MS:m/z＝329[M+H]+ To a solution of tert-butyl 4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazine-1-carboxylate (10-6) (170 mg, 0.397 mmol, 1.0 eq.) in EA (5 mL) was added 4M HCl in dioxane (2.5 mL) at room temperature. Stirring was continued for 16 hours and LCMS showed that the reaction was complete. EA (20 mL) was added to the mixture and stirred for 5 minutes before filtering. The filter cake was dried to give 1-(1-methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (11-7) (100 mg). MS: m/z=329[M+H] +

第七步：化合物11的制备Step 7: Preparation of compound 11

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(103毫克，0.21毫摩尔，1.0eq.)和1-(1-甲基-6-(哌嗪-1-基)-1H-吲唑-3-基)二氢嘧啶-2，4(1H，3H)-二酮(11-7)(70毫克，0.21毫摩尔，1.0eq.)的DCM(5毫升)中加入TEA(216毫克，2.13毫摩尔，10.0eq.)和MgSO4(256毫克，2.13毫摩尔，10.0eq.)。在25℃下搅拌2小时后，加入NaBH(OAc)3(113mg，0.53mmol，2.5eq.)。在25℃下搅拌16小时，加入DCM(20mL)和水(20mL)。过滤混合物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-((4-(3-(2，4-二氧代四氢嘧啶-1(2H)-基)-1-甲基-1H-吲唑-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酰胺(11)(60毫克，产率35％)。MS:m/z＝795[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (103 mg, 0.21 mmol, 1.0 eq.) and 1-(1-methyl-6-(piperazin-1-yl)-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (11-7) (70 mg, 0.21 mmol, 1.0 eq.) in DCM (5 ml) was added TEA (216 mg, 2.13 mmol, 10.0 eq.) and MgSO 4 (256 mg, 2.13 mmol, 10.0 eq.) at room temperature. After stirring at 25° C. for 2 hours, NaBH(OAc) 3 (113 mg, 0.53 mmol, 2.5 eq.) was added. The mixture was stirred at 25°C for 16 hours, and DCM (20 mL) and water (20 mL) were added. The mixture was filtered to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-((4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (11) (60 mg, 35% yield). MS: m/z=795[M+H] +

1H NMR(400MHz,DMSO-d6)δ10.48(s,1H),9.04(d,J＝3.2Hz,1H),8.62(d,J＝7.6Hz,1H),8.26(d,J＝8.0Hz,1H),7.97(d,J＝8.0Hz,1H),7.73-7.65(m,2H),7.43(d,J＝9.2Hz,1H),7.33(d,J＝8.8Hz, 1H),6.92-6.89(m,2H),6.80(s,1H),4.73-4.68(m,1H),3.89-3.81(m,5H),3.30(s,10H),3.21(bs,3H),2.83-2.69(m,3H),2.51(bs,3H),2.24-2.19(m,3H),1.96-1.92(m,2H),1.81-1.77(m,2H),1.68-1.55(m,3H),1.23-1.13(m,2H). 1 H NMR (400MHz, DMSO-d6) δ10.48(s,1H),9.04(d,J＝3.2Hz,1H),8.62(d,J＝7.6Hz,1H),8.26(d,J＝ 8.0Hz,1H),7.97(d,J＝8.0Hz,1H),7.73-7.65(m,2H),7.43(d,J＝9.2Hz,1H),7.33(d,J＝8.8Hz, 1H),6.92-6.89(m,2H),6.80(s,1H),4.73-4.68(m,1H),3.89-3.81(m,5H),3.30(s,10H),3.21(bs,3H) ,2.83-2.69( m,3H),2.51(bs,3H),2.24-2.19(m,3H),1.96-1.92(m,2H),1.81-1.77(m,2H),1.68-1.55(m,3H),1.23- 1.13(m,2H).

实施例12化合物12的制备
Example 12 Preparation of Compound 12

第一步：化合物12-1的制备Step 1: Preparation of compound 12-1

室温下，向1-溴-3-氯-5-硝基苯(100毫克，0.42毫摩尔，1eq.)的toluene(2mL)中加入哌嗪-1-羧酸叔丁酯(94.5毫克，0.51毫摩尔，1.2eq.)，Pd2(dba)3(19.3毫克，0.02毫摩尔，0.05eq.)，BINAP(26.3毫克，0.042毫摩尔，0.10eq.)和NaOtBu(56.9毫克，0.59毫摩尔，1.4eq.)。在100℃下搅拌4小时。冷却后，将反应混合物通过硅藻土垫过滤，滤液浓缩。经快速色谱(己烷/乙酸乙酯＝18：1)纯化，得到4-(3-氯-5-硝基苯基)哌嗪-1-羧酸叔丁酯(12-1)(195mg)。MS:m/z＝267[M+H]+ To 1-bromo-3-chloro-5-nitrobenzene (100 mg, 0.42 mmol, 1 eq.) in toluene (2 mL) were added tert-butyl piperazine-1-carboxylate (94.5 mg, 0.51 mmol, 1.2 eq.), Pd 2 (dba) 3 (19.3 mg, 0.02 mmol, 0.05 eq.), BINAP (26.3 mg, 0.042 mmol, 0.10 eq.) and NaOtBu (56.9 mg, 0.59 mmol, 1.4 eq.) at room temperature. The mixture was stirred at 100°C for 4 hours. After cooling, the reaction mixture was filtered through a celite pad and the filtrate was concentrated. Purification by flash chromatography (hexane/ethyl acetate = 18:1) gave tert-butyl 4-(3-chloro-5-nitrophenyl)piperazine-1-carboxylate (12-1) (195 mg). MS: m/z = 267 [M + H] +

第二步：化合物12-2的制备Step 2: Preparation of compound 12-2

室温下，向4-(3-氯-5-硝基苯基)哌嗪-1-羧酸叔丁酯(12-1)(190mg，0.55mmol，1.0eq.)的MeOH(40mL)溶液中加入PtO2(20mg，0.1w.)。在H2气氛下搅拌4小时后，过滤混合物，滤液浓缩，得到4-(3-氨基-5-氯苯基)哌嗪-1-羧酸叔丁酯(12-2)(150毫克)。MS:m/z＝312[M+H]+ PtO 2 (20 mg, 0.1 w.) was added to a solution of tert-butyl 4-(3-chloro-5-nitrophenyl)piperazine-1-carboxylate (12-1) (190 mg, 0.55 mmol, 1.0 eq.) in MeOH (40 mL) at room temperature. After stirring for 4 hours under H 2 atmosphere, the mixture was filtered and the filtrate was concentrated to give tert-butyl 4-(3-amino-5-chlorophenyl)piperazine-1-carboxylate (12-2) (150 mg). MS: m/z=312 [M+H] +

第三步：化合物12-3的制备Step 3: Preparation of compound 12-3

室温下，向4-(3-氨基-5-氯苯基)哌嗪-1-羧酸叔丁酯(12-2)(150mg，0.48mmol，1.0eq.)的DMF(2mL)溶液中加入3-溴哌啶-2，6-二酮(183mg，0.96mmol，2.0eq.)和NaHCO3(121mg，1.44mmol，3.0eq.)。在70℃下搅拌后，冷至室温，通过反相色谱(TFA缓冲液)纯化反应混合物，得到4-(3-氯-5-((2，6-二氧代哌啶-3-基)氨基)苯基)哌嗪-1-羧酸叔丁酯(12-3)(100mg)。MS:m/z＝423[M+H]+ 3-Bromopiperidine-2,6-dione (183 mg, 0.96 mmol, 2.0 eq.) and NaHCO 3 (121 mg, 1.44 mmol, 3.0 eq.) were added to a solution of tert-butyl 4-(3-amino-5-chlorophenyl)piperazine- 1 -carboxylate (12-2) (150 mg, 0.48 mmol, 1.0 eq.) in DMF (2 mL) at room temperature. After stirring at 70°C, the mixture was cooled to room temperature and the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give tert-butyl 4-(3-chloro-5-((2,6-dioxopiperidin-3-yl)amino)phenyl)piperazine-1-carboxylate (12-3) (100 mg). MS: m/z=423 [M+H] +

第四步：化合物12-4的制备Step 4: Preparation of compound 12-4

室温下，向4-(3-氯-5-((2，6-二氧代哌啶-3-基氨基)苯基)哌嗪-1-羧酸叔丁酯(12-3)(100mg，0.23mmol，1.0eq.)的DCM(3mL)溶液中加入4M HCl in EA(3mL)。在室温下搅拌2小时，浓缩，得到3-((3-氯-5-(哌嗪-1-基)苯基)氨基)哌啶-2，6-二酮盐酸盐(12-4)(70毫克，粗品)。MS:m/z＝323[M+H]+ To a solution of tert-butyl 4-(3-chloro-5-((2,6-dioxopiperidin-3-ylamino)phenyl)piperazine-1-carboxylate (12-3) (100 mg, 0.23 mmol, 1.0 eq.) in DCM (3 mL) was added 4M HCl in EA (3 mL) at room temperature. The mixture was stirred at room temperature for 2 hours and concentrated to give 3-((3-chloro-5-(piperazin-1-yl)phenyl)amino)piperidine-2,6-dione hydrochloride (12-4) (70 mg, crude). MS: m/z=323 [M+H] +

第五步：化合物12的制备Step 5: Preparation of compound 12

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(37mg，0.077mmol，1eq.)的DCM(1mL)溶液中加入3-((3-氯-5-(哌嗪-1-基苯基)氨基)哌啶-2，6-二酮(12-4)(25.0毫克，0.077毫摩尔，1.0eq.)，TEA(3滴)和MgSO4(93毫克， 0.07毫摩尔，10.0eq.)。室温下搅拌30分钟后，加入NaBH(OAc)3(16mg，0.077mmol，1.0eq.)。在室温下搅拌1小时后，通过反相色谱(TFA缓冲液)纯化反应混合物，得到4-(4-(3-氯-5-((2，6-二氧代哌啶-3-基)氨基)苯基)哌嗪-1-基)甲基)哌啶-1-基)-N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)苯甲酰胺(12)(17.6mg)。MS:m/z＝789[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (37 mg, 0.077 mmol, 1 eq.) in DCM (1 mL) at room temperature were added 3-((3-chloro-5-(piperazin-1-ylphenyl)amino)piperidine-2,6-dione (12-4) (25.0 mg, 0.077 mmol, 1.0 eq.), TEA (3 drops) and MgSO 4 (93 mg, 0.077 mmol, 1 eq.). 0.07 mmol, 10.0 eq.). After stirring at room temperature for 30 minutes, NaBH(OAc) 3 (16 mg, 0.077 mmol, 1.0 eq.) was added. After stirring at room temperature for 1 hour, the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give 4-(4-(3-chloro-5-((2,6-dioxopiperidin-3-yl)amino)phenyl)piperazin-1-yl)methyl)piperidin-1-yl)-N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)benzamide (12) (17.6 mg). MS: m/z=789[M+H] +

1H NMR(400MHz,DMSO-d6)δ10.78(s,1H),9.37(s,1H),9.05(dd,J＝4.4,1.7Hz,1H),8.62(dd,J＝8.4,1.8Hz,1H),8.27(d,J＝8.4Hz,1H),7.99(d,J＝7.6Hz,1H),7.74(d,J＝8.6Hz,2H),7.67(dd,J＝8.4,4.2Hz,1H),7.33(d,J＝8.4Hz,1H),6.94(d,J＝8.8Hz,2H),6.27–6.19(m,3H),4.76–4.69(m,2H),4.37(dd,J＝11.8,4.9Hz,2H),3.86(d,J＝10.2Hz,4H),3.74(d,J＝12.2Hz,3H),3.57(d,J＝11.0Hz,3H),3.05–2.99(m,2H),2.85–2.74(m,2H),2.58(d,J＝4.2Hz,1H),2.55–2.51(m,1H),2.25–2.19(m,2H),2.07–2.02(m,2H),1.97–1.89(m,2H),1.85–1.78(m,3H),1.68–1.62(m,2H),1.58–1.53(m,1H),1.31–1.24(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ10.78(s,1H),9.37(s,1H),9.05(dd,J＝4.4,1.7Hz,1H),8.62(dd,J＝8.4,1.8Hz,1H),8.27(d,J ＝8.4Hz,1H),7.99(d,J＝7.6Hz,1H),7.74(d,J＝8. 6Hz,2H),7.67(dd,J＝8.4,4.2Hz,1H),7.33(d,J＝8.4Hz,1H),6.94(d,J＝8.8Hz,2H),6.27–6.19(m,3H ),4.76–4.69(m,2H),4.37(dd,J＝11.8,4.9Hz ,2H),3.86(d,J＝10.2Hz,4H),3.74(d,J＝12.2Hz,3H),3.57(d,J＝11.0Hz, 3H),3.05–2.99(m,2H),2.85–2.74(m,2H),2.58(d,J＝4.2Hz,1H),2.55–2 .51(m,1H),2.25–2.19(m,2H),2.07–2.02(m,2H),1.97–1.89(m,2H),1.8 5–1.78(m,3H),1.68–1.62(m,2H),1.58–1.53(m,1H),1.31–1.24(m,2H).

实施例13化合物13的制备
Example 13 Preparation of Compound 13

第一步：化合物13-1的制备Step 1: Preparation of compound 13-1

室温下，向4-羟基哌啶-1-羧酸苄酯溶液(3.0g，21.1mmol，1eq.)的THF(30mL)溶液中加入4-硝基-1H-吡唑(2.39克，21.1毫摩尔，1.0eq.)、DIAD(5.13克、25.4毫摩尔、1.2eq.)和PPh3(6.6克、25.4毫摩尔、1.2eq.)。在室温下搅拌12h，加水(30mL)，并用EA(3x 30mL)萃取。将有机层用Na2SO4干燥并浓缩。残余物在用PE/EA(10：1至3：1)洗脱的硅胶上色谱纯化，得到4-(4-硝基-1H-吡唑-1-基)哌啶-1-羧酸苄酯(13-1)(3.3g)。MS:m/z＝331[M+H]+ To a solution of 4-hydroxypiperidine-1-carboxylic acid benzyl ester (3.0 g, 21.1 mmol, 1 eq.) in THF (30 mL) at room temperature, 4-nitro-1H-pyrazole (2.39 g, 21.1 mmol, 1.0 eq.), DIAD (5.13 g, 25.4 mmol, 1.2 eq.) and PPh 3 (6.6 g, 25.4 mmol, 1.2 eq.) were added. The mixture was stirred at room temperature for 12 h, water (30 mL) was added, and extracted with EA (3 x 30 mL). The organic layer was dried over Na 2 SO 4 and concentrated. The residue was chromatographed on silica gel eluted with PE/EA (10:1 to 3:1) to give 4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid benzyl ester (13-1) (3.3 g). MS: m/z=331 [M+H] +

第二步：化合物13-2的制备Step 2: Preparation of compound 13-2

室温下，向4-(4-硝基-1H-吡唑-1-基)哌啶-1-羧酸苄酯(13-1)(3.3克，16.4毫摩尔，1.0eq.)的甲醇(40毫升)溶液中加入PtO2(330毫克，0.1毫)。在H2气氛下搅拌4小时后，过滤混合物，滤液浓缩得到4-(4-氨基-1H-吡唑-1-基)哌啶-1-羧酸酯(13-2)(3.0克)。MS:m/z＝301[M+H]+ PtO 2 (330 mg, 0.1 mmol) was added to a solution of 4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylic acid benzyl ester (13-1) (3.3 g, 16.4 mmol, 1.0 eq.) in methanol (40 ml) at room temperature. After stirring for 4 hours under H 2 atmosphere, the mixture was filtered and the filtrate was concentrated to give 4-(4-amino-1H-pyrazol-1-yl)piperidine-1-carboxylic acid ester (13-2) (3.0 g). MS: m/z=301 [M+H] +

第三步：化合物13-3的制备Step 3: Preparation of compound 13-3

室温下，将乳酸(5.0g，56.3mmol，5.0eq)和DBU(8.5g，56.3mmol，5.0eq.)的混合物在25℃下搅拌16h，然后加入4-(4-氨基-1H-吡唑-1-基)哌啶-1-羧酸酯(13-2)(3.0g，11.27mmol，1.0eq)和丙烯酸甲酯(30mL)。在室温下搅拌16小时后，加入EA(50mL)和水(50mL)。浓缩有机层，通过反相色谱(TFA缓冲液)纯化反应混合物，得到4-(4-(3-甲氧基-3-氧代丙基)氨基)-1H-吡唑-1- 基)哌啶-1-羧酸苄酯(13-3)(2.28克)。MS:m/z＝387[M+H]+ At room temperature, a mixture of lactic acid (5.0 g, 56.3 mmol, 5.0 eq) and DBU (8.5 g, 56.3 mmol, 5.0 eq.) was stirred at 25 ° C for 16 h, and then 4-(4-amino-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-2) (3.0 g, 11.27 mmol, 1.0 eq) and methyl acrylate (30 mL) were added. After stirring at room temperature for 16 hours, EA (50 mL) and water (50 mL) were added. The organic layer was concentrated and the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give 4-(4-(3-methoxy-3-oxopropyl)amino)-1H-pyrazole-1- Benzyl)piperidine-1-carboxylate (13-3) (2.28 g). MS: m/z = 387 [M+H] +

第四步：化合物13-4的制备Step 4: Preparation of compound 13-4

室温下，向4-(4-(3-甲氧基-3-氧代丙基)氨基)-1H-吡唑-1-基)哌啶-1-羧酸苄酯(13-3)(620毫克，1.6毫摩尔，1.0eq.)的ACOH(6.2毫升)溶液中加入KCNO(260毫克，3.2毫摩尔，2.0eq.)。在25℃下搅拌16h后，加入EA(20mL)和水(20mL)。浓缩有机层，残余物在用EA/MeOH(10：1)洗脱的硅胶上色谱纯化，得到4-(4-(1-(3-甲氧基-3-氧代丙基)脲基)-1H-吡唑-1-基)哌啶-1-羧酸苄酯(13-4)(470mg)。MS:m/z＝430[M+H]+ To a solution of benzyl 4-(4-(3-methoxy-3-oxopropyl)amino)-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-3) (620 mg, 1.6 mmol, 1.0 eq.) in ACOH (6.2 mL) was added KCNO (260 mg, 3.2 mmol, 2.0 eq.) at room temperature. After stirring at 25°C for 16 h, EA (20 mL) and water (20 mL) were added. The organic layer was concentrated and the residue was purified by chromatography on silica gel eluted with EA/MeOH (10:1) to give benzyl 4-(4-(1-(3-methoxy-3-oxopropyl)ureido)-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-4) (470 mg). MS: m/z=430 [M+H] +

第五步：化合物13-5的制备Step 5: Preparation of compound 13-5

室温下，向4-(4-(1-(3-(3-甲氧基-3-氧代丙基)脲基)-1H-吡唑-1-基)哌啶-1-羧酸苄酯(13-4)(420mg，0.97mmol，1.0eq.)的MeCN(12mL)溶液中加入Triton-B(409mg，40％，2.44mmol，2.5eq.)。在25℃下搅拌16h后，加入EA(50mL)和水(50mL)。将有机层浓缩得到4-(4-(2，4-二氧代四氢嘧啶-1(2H)-基)-1H-吡唑-1-基)哌啶-1-羧酸苄酯(13-5)(450毫克)。MS:m/z＝398[M+H]+ Triton-B (409 mg, 40%, 2.44 mmol, 2.5 eq.) was added to a solution of benzyl 4-(4-(1-(3-(3-methoxy-3-oxopropyl)ureido)-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-4) (420 mg, 0.97 mmol, 1.0 eq.) in MeCN (12 mL) at room temperature. After stirring at 25°C for 16 h, EA (50 mL) and water (50 mL) were added. The organic layer was concentrated to give benzyl 4-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-5) (450 mg). MS: m/z=398 [M+H] +

第六步：化合物13-6的制备Step 6: Preparation of compound 13-6

室温下，向4-(4-(2，4-二氧代四氢嘧啶-1(2H)-基)-1H-吡唑-1-基)哌啶-1-羧酸苄酯(13-5)(450毫克，1.13毫摩尔，1.0eq.)的溶液中加入MeOH(40毫升)中PtO2(45毫克，0.1瓦)。在H2气氛下搅拌4小时后，过滤混合物，滤液浓缩，得到1-(1-(哌啶-4-基)-1H-吡唑-4-基)二氢嘧啶-2，4(1H，3H)-二酮(13-6)(300毫克)。MS:m/z＝283[M+H]+ To a solution of benzyl 4-(4-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-5) (450 mg, 1.13 mmol, 1.0 eq.) was added PtO 2 (45 mg, 0.1 W) in MeOH (40 mL) at room temperature. After stirring for 4 h under H 2 atmosphere, the mixture was filtered and the filtrate was concentrated to give 1-(1-(piperidin-4-yl)-1H-pyrazol-4-yl)dihydropyrimidine-2,4(1H,3H)-dione (13-6) (300 mg). MS: m/z=283[M+H] +

第七步：化合物13的制备Step 7: Preparation of compound 13

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(90mg，0.19mmol，1.0eq.)和1-(1-((哌啶-4-基)-1H-吡唑-4-基)二氢嘧啶-2，4(1H，3H)-二氢嘧啶-2，4(1H，3H)-二酮(13-6)(50mg，0.19mmol，1.0eq.)的DCM(5mL)溶液中加入TEA(4滴)。在25℃下搅拌2小时，然后加入NaBH(OAc)3(120.1mg，0.5mmol，3.0eq.)。将反应混合物在25℃下搅拌16小时，加入DCM(20mL)和水(20mL)。过滤混合物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-((4-(3-(2，4-二氧代四氢嘧啶-1(2H)-基)-1-甲基-1H-吲唑-6-基)哌嗪-1-基)甲基)哌啶-1-基)苯甲酰胺(13)(21.4mg)。MS:m/z＝730[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (90 mg, 0.19 mmol, 1.0 eq.) and 1-(1-((piperidin-4-yl)-1H-pyrazol-4-yl)dihydropyrimidine-2,4(1H,3H)-dihydropyrimidine-2,4(1H,3H)-dione (13-6) (50 mg, 0.19 mmol, 1.0 eq.) in DCM (5 mL) was added TEA (4 drops) at room temperature. The mixture was stirred at 25°C for 2 hours, and then NaBH(OAc) was added . (120.1 mg, 0.5 mmol, 3.0 eq.). The reaction mixture was stirred at 25°C for 16 hours, and DCM (20 mL) and water (20 mL) were added. The mixture was filtered to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-((4-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)benzamide (13) (21.4 mg). MS: m/z=730 [M+H] +

1H NMR(400MHz,DMSO-d6)δ10.40(s,1H),9.32(brs,1H),9.08(dd,J＝4.2,1.7Hz,1H),8.65(dd,J＝8.5,1.7Hz,1H),8.29(d,J＝8.3Hz,1H),8.02(d,J＝7.6Hz,1H),7.99(s,1H),7.76(d,J＝8.7Hz,2H),7.69(dd,J＝8.5,4.3Hz,1H),7.36(d,J＝8.5Hz,1H),6.97(d,J＝8.9Hz,2H),4.74(brs,1H),4.47(brs,1H),3.89(d,J＝11.1Hz,3H),3.77(t,J＝6.8Hz,3H),3.68(d,J＝11.1Hz,2H),3.18–3.00(m,4H),2.81(t,J＝11.5Hz,2H),2.70(t,J＝6.8Hz,2H),2.30–2.19(m,5H),2.07(brs,1H),2.01–1.89(m,2H),1.86(d,J＝12.6Hz,2H),1.69–1.55(m,4H),1.39–1.20(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ10.40(s,1H),9.32(brs,1H),9.08(dd,J＝4.2,1.7Hz,1H),8.65(dd,J＝8.5,1.7Hz,1H),8.29(d,J ＝8.3Hz,1H),8.02(d,J＝7.6Hz,1 H),7.99(s,1H),7.76(d,J＝8.7Hz,2H),7.69(dd,J＝8.5,4.3Hz,1H),7.36(d,J＝8.5Hz,1H),6.97( d,J＝8.9Hz,2H),4.74(brs,1H),4.4 7(brs,1H),3.89(d,J＝11.1Hz,3H),3.77(t,J＝6.8Hz,3H),3.68(d,J＝11.1Hz,2H),3.18–3.00(m,4H ),2.81(t,J＝11.5Hz,2H),2.70( t,J＝6.8Hz,2H),2.30–2.19(m,5H),2.07(brs,1H),2.01–1.89(m,2H),1.86(d,J＝12.6Hz,2H),1.69–1.55 (m,4H),1.39–1.20(m,2H).

实施例14化合物14的制备
Example 14 Preparation of Compound 14

第一步：化合物14-1的制备Step 1: Preparation of compound 14-1

室温下，向4-羟基哌啶-1-羧酸叔丁酯溶液(300mg，1.49mmol，1eq.)的THF(3mL)溶液中加入4-硝基-1H-吡唑(168毫克，1.49毫摩尔，1.0eq.)，DIAD(361毫克，1.79毫摩尔，1.2eq.)和PPh3(469毫克，1.79毫摩尔，1.2eq.)。在室温下搅拌4h后，加入水(20mL)稀释混合物并用EA(3x 20mL)萃取。将有机层用Na2SO4干燥并浓缩。残余物用柱层析纯化(PE/EA(10：1至3：1))，得到4-(4-硝基-1H-吡唑-1-基)哌啶-1-羧酸叔丁酯(14-1)(100mg)。MS:m/z＝297[M+H]+ To a solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (300 mg, 1.49 mmol, 1 eq.) in THF (3 mL) at room temperature were added 4-nitro-1H-pyrazole (168 mg, 1.49 mmol, 1.0 eq.), DIAD (361 mg, 1.79 mmol, 1.2 eq.) and PPh 3 (469 mg, 1.79 mmol, 1.2 eq.). After stirring at room temperature for 4 h, water (20 mL) was added to dilute the mixture and extracted with EA (3 x 20 mL). The organic layer was dried over Na 2 SO 4 and concentrated. The residue was purified by column chromatography (PE/EA (10:1 to 3:1)) to give tert-butyl 4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (14-1) (100 mg). MS: m/z=297 [M+H] +

第二步：化合物14-2的制备Step 2: Preparation of compound 14-2

室温下，向4-(4-硝基-1H-吡唑-1-基)哌啶-1-羧酸叔丁酯(13-1)(100mg，0.55mmol，1.0eq.)的MeOH(40mL)溶液中加入PtO2(10mg，0.1w.)。将所得混合物在H2气氛下搅拌4小时后，过滤，滤液浓缩，得到4-(4-氨基-1H-吡唑-1-基)哌啶-1-羧酸叔丁酯(14-2)(70mg，粗品)。MS:m/z＝267[M+H]+ PtO 2 (10 mg, 0.1 w.) was added to a solution of tert-butyl 4-(4-nitro-1H-pyrazol-1-yl)piperidine-1-carboxylate (13-1) (100 mg, 0.55 mmol, 1.0 eq.) in MeOH (40 mL) at room temperature. The resulting mixture was stirred under H 2 atmosphere for 4 hours, filtered, and the filtrate was concentrated to give tert-butyl 4-(4-amino-1H-pyrazol-1-yl)piperidine-1-carboxylate (14-2) (70 mg, crude product). MS: m/z=267 [M+H] +

第三步：化合物14-3的制备Step 3: Preparation of compound 14-3

室温下，向4-(4-氨基-1H-吡唑-1-基)哌啶-1-羧酸叔丁酯(14-2)(70mg，0.48mmol，1.0eq.)的DMF(2mL)溶液中加入3-溴哌啶-2，6-二酮(183mg，0.96mmol，2.0eq.)和NaHCO3(121mg，1.44mmol，3.0eq.)。在70℃下搅拌1小时后，冷至室温，通过反相色谱(TFA缓冲液)纯化反应混合物，得到4-(4-((2，6-二氧代哌啶-3-基氨基)-1H-吡唑-1-基)哌啶-1-羧酸叔丁酯(14-3)(50mg)。MS:m/z＝378[M+H]+ To a solution of tert-butyl 4-(4-amino-1H-pyrazol-1-yl)piperidine-1-carboxylate (14-2) (70 mg, 0.48 mmol, 1.0 eq.) in DMF (2 mL) at room temperature were added 3-bromopiperidine-2,6-dione (183 mg, 0.96 mmol, 2.0 eq.) and NaHCO 3 (121 mg, 1.44 mmol, 3.0 eq.). After stirring at 70° C. for 1 hour, the mixture was cooled to room temperature and the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give tert-butyl 4-(4-((2,6-dioxopiperidin-3-ylamino)-1H-pyrazol-1-yl)piperidine-1-carboxylate (14-3) (50 mg). MS: m/z=378 [M+H] +

第四步：化合物14-4的制备Step 4: Preparation of compound 14-4

室温下，向4-(4-(((2，6-二氧代哌啶-3-基氨基)-1H-吡唑-1-基)哌啶-1-羧酸叔丁酯(14-3)(50.0mg，0.23毫摩尔，1.0eq.)的DCM(3mL)溶液中加入4M HCl的EA(3mL)溶液。室温下搅拌2小时后，浓缩，得到化合物14-4(25毫克)。MS:m/z＝278[M+H]+ To a solution of tert-butyl 4-(4-(((2,6-dioxopiperidin-3-ylamino)-1H-pyrazol-1-yl)piperidine-1-carboxylate (14-3) (50.0 mg, 0.23 mmol, 1.0 eq.) in DCM (3 mL) was added 4M HCl in EA (3 mL) at room temperature. After stirring at room temperature for 2 hours, the mixture was concentrated to give compound 14-4 (25 mg). MS: m/z=278[M+H] +

第五步：化合物14的制备Step 5: Preparation of compound 14

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(43mg，0.09mmol，1eq.)的DCM(1mL)溶液中加入化合物14-4(25.0毫克，0.09毫摩尔，1.0eq.)，TEA(3滴)和MgSO4(108毫克，0.07毫摩尔，10.0eq.)。在室温下搅拌30分钟，然后加入NaBH(OAc)3(19mg，0.09mmol，1.0eq.)。将反应混合物在室温下搅拌1小时后，通过反相色谱(TFA缓冲液)纯化反应混合物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-((4-((2，6-二氧代哌啶-3-基)氨基)-1H-吡唑-1-基)哌啶-1-基)甲基)哌啶-1-基)苯甲酰胺(14)(6.1mg)。MS:m/z＝744[M+H]+ Compound 14-4 (25.0 mg, 0.09 mmol, 1.0 eq.), TEA (3 drops) and MgSO4 (108 mg, 0.07 mmol, 10.0 eq.) were added to a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (43 mg, 0.09 mmol, 1 eq.) in DCM (1 mL) at room temperature. The mixture was stirred at room temperature for 30 minutes, and then NaBH(OAc) 3 (19 mg, 0.09 mmol, 1.0 eq.) was added. After the reaction mixture was stirred at room temperature for 1 hour, the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-((4-((2,6-dioxopiperidin-3-yl)amino)-1H-pyrazol-1-yl)piperidin-1-yl)methyl)piperidin-1-yl)benzamide (14) (6.1 mg). MS: m/z=744 [M+H] +

1H NMR(400MHz,DMSO-d6)δ10.77(s,1H),9.05(dd,J＝4.2,1.8Hz,2H),8.62(dd,J＝8.4,1.8Hz, 1H),8.27(d,J＝8.4Hz,1H),8.02–7.97(m,1H),7.74(d,J＝8.4Hz,2H),7.67(dd,J＝8.4,4.3Hz,1H),7.33(d,J＝8.4Hz,1H),6.94(d,J＝9.0Hz,2H),4.75–4.68(m,1H),4.32–4.25(m,1H),3.92–3.77(m,6H),3.15–2.96(m,4H),2.78(t,J＝12.2Hz,2H),2.26–2.11(m,6H),1.97–1.88(m,2H),1.85–1.78(m,2H),1.70–1.49(m,4H),1.31–1.20(m,2H). 1 H NMR (400MHz, DMSO-d 6 ) δ10.77 (s, 1H), 9.05 (dd, J＝4.2, 1.8Hz, 2H), 8.62 (dd, J＝8.4, 1.8Hz, 1H),8.27(d,J＝8.4Hz,1H),8.02–7.97(m,1H),7.74(d,J＝8.4Hz,2H),7.67(dd,J＝8.4 ,4.3Hz,1H),7.33(d,J＝8.4Hz,1H),6.94(d,J＝9.0Hz,2H),4.75–4.68(m,1H),4.32–4 .25(m,1H),3.92–3.77(m,6H),3.15–2.96(m,4H),2.78(t,J＝12.2Hz,2H),2.26–2.11 (m,6H),1.97–1.88(m,2H),1.85–1.78(m,2H),1.70–1.49(m,4H),1.31–1.20(m,2H).

实施例15化合物15的制备
Example 15 Preparation of Compound 15

第一步：化合物15-1的制备Step 1: Preparation of compound 15-1

室温下，向6-氯哒嗪-3-羧酸(2g，12.6mmol，1.0eq.)，DMAP(776mg，6.3mmol，0.5eq.)的THF(50mL)中，加入Boc2O(3.6g，16.4mmol，1.3eq.)，在50℃下搅拌6小时后，加日H2O(400mL)，用EA(100mL*2)萃取，合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，真空浓缩。残余物用硅胶色谱(PE：EA(1：1))纯化，得到6-氯哒嗪-3-羧酸叔丁酯(15-1)(2.2g)。MS:m/z＝158[M+H-56]+ At room temperature, Boc 2 O (3.6 g, 16.4 mmol, 1.3 eq.) was added to a solution of 6-chloropyridazine-3-carboxylic acid (2 g, 12.6 mmol, 1.0 eq.) and DMAP (776 mg, 6.3 mmol, 0.5 eq.) in THF (50 mL). After stirring at 50°C for 6 hours, H 2 O (400 mL) was added and the mixture was extracted with EA (100 mL*2). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo. The residue was purified by silica gel chromatography (PE:EA (1:1)) to give tert-butyl 6-chloropyridazine-3-carboxylate (15-1) (2.2 g). MS: m/z=158[M+H-56] +

第二步：化合物15-2的制备Step 2: Preparation of compound 15-2

室温下，向6-氯哒嗪-3-羧酸叔丁酯(15-1)(1克，4.66毫摩尔，1.0eq.)，哌啶-4-基甲醇(1.33克，9.31毫摩尔，2.0eq.)的1,4-dioxane(25mL)中加入DIPEA(1.5毫升，9.31毫摩尔，2.0eq.)。在100℃搅拌16小时后，冷至室温，浓缩。残余物用柱色谱PE：EA＝1：3纯化，得到6-(4-(羟甲基)哌啶-1-基)哒嗪-3-羧酸叔丁酯(15-2)(900mg)。MS:m/z＝293[M+H-56]+ At room temperature, DIPEA (1.5 mL, 9.31 mmol, 2.0 eq.) was added to tert-butyl 6-chloropyridazine-3-carboxylate (15-1) (1 g, 4.66 mmol, 1.0 eq.), piperidin-4-ylmethanol (1.33 g, 9.31 mmol, 2.0 eq.) in 1,4-dioxane (25 mL). After stirring at 100°C for 16 hours, the mixture was cooled to room temperature and concentrated. The residue was purified by column chromatography PE:EA=1:3 to obtain tert-butyl 6-(4-(hydroxymethyl)piperidin-1-yl)pyridazine-3-carboxylate (15-2) (900 mg). MS: m/z=293[M+H-56] +

第三步：化合物15-3的制备Step 3: Preparation of compound 15-3

室温下，将化合物15-2(100mg，0.34mmol，1.0eq.)和DMP(173mg，0.41mmol，1.2eq.)在DCM(2mL)中的混合物室温搅拌2小时。加入饱和Na2S2O3(50mL)稀释混合物，并分别用饱和NaHCO3(50mL)和H2O(50mL)洗涤，水层用DCM(10mL)萃取，合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，真空浓缩，得6-(4-甲酰基哌啶-1-基)哒嗪-3-羧酸叔丁酯(15-3)(80mg)。A mixture of compound 15-2 (100 mg, 0.34 mmol, 1.0 eq.) and DMP (173 mg, 0.41 mmol, 1.2 eq.) in DCM (2 mL) was stirred at room temperature for 2 hours. Saturated Na 2 S 2 O 3 (50 mL) was added to dilute the mixture, and the mixture was washed with saturated NaHCO 3 (50 mL) and H 2 O (50 mL), respectively. The aqueous layer was extracted with DCM (10 mL), and the combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to obtain tert-butyl 6-(4-formylpiperidin-1-yl)pyridazine-3-carboxylate (15-3) (80 mg).

第四步：化合物15-4的制备Step 4: Preparation of compound 15-4

室温下，将化合物15-3(60毫克，0.21毫摩尔，1.0eq.)，3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮4-5(69毫克，0.21毫摩尔，1.0eq.)，TEA(90μL，0.62毫摩尔，3.0eq.)，MgSO4 (20毫克)在DCM(2mL)的化合物，温搅拌16小时。然后加入NaBH(OAc)3(51毫克，0.25毫摩尔，1.2eq.)并在室温搅拌2小时后，加入H2O(10mL)，并用DCM(10mL*2)萃取。合并有机层，真空浓缩。残余物用硅胶色谱(PE：EA(1：1))纯化，得到6-(4-((2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哒啶-1-基)哒嗪-3-羧酸叔丁基酯(15-4)(100mg)。MS:m/z＝616[M+H]+ At room temperature, compound 15-3 (60 mg, 0.21 mmol, 1.0 eq.), 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione 4-5 (69 mg, 0.21 mmol, 1.0 eq.), TEA (90 μL, 0.62 mmol, 3.0 eq.), MgSO 4 (20 mg) of the compound in DCM (2 mL) was stirred at room temperature for 16 hours. Then NaBH(OAc) 3 (51 mg, 0.25 mmol, 1.2 eq.) was added and stirred at room temperature for 2 hours, then H 2 O (10 mL) was added and extracted with DCM (10 mL*2). The organic layers were combined and concentrated in vacuo. The residue was purified by silica gel chromatography (PE: EA (1: 1)) to give 6-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)pyridin-1-yl)pyridazine-3-carboxylic acid tert-butyl ester (15-4) (100 mg). MS: m/z=616[M+H] +

第五步：化合物15-5的制备Step 5: Preparation of compound 15-5

室温下，向化合物15-4(100毫克，0.16毫摩尔，1.0eq.)的DCM(1mL)溶液中加入TFA(0.5毫升)，搅拌3小时后，浓缩，得到6-(4-((2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哒啶-1-基)哒嗪-3-羧酸(15-5)(90毫克)。MS:m/z＝561[M+H]+ At room temperature, TFA (0.5 ml) was added to a DCM (1 mL) solution of compound 15-4 (100 mg, 0.16 mmol, 1.0 eq.), and the mixture was stirred for 3 hours and concentrated to give 6-(4-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)pyridin-1-yl)pyridazine-3-carboxylic acid (15-5) (90 mg). MS: m/z=561[M+H] +

第六步：化合物15的制备Step 6: Preparation of compound 15

室温下，向化合物15-5(40毫克，0.071毫摩尔，1.0eq.)，HATU(33毫克，0.082毫摩尔，1.2等量)和DIEA(27毫克，0.231毫摩尔，3.0eq.)的DMF(2mL)中加入5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(20毫克，0.071毫摩尔，1.0eq.)。温搅拌16小时后，加入H2O(10mL)，并用DCM(10mL*2)萃取，合并有机层，真空浓缩。残余物通过反相色谱(TFA缓冲液)纯化得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-甲酰胺(15)(21mg)。MS：m/z＝810[M+H]+ To a solution of compound 15-5 (40 mg, 0.071 mmol, 1.0 eq.), HATU (33 mg, 0.082 mmol, 1.2 eq.) and DIEA (27 mg, 0.231 mmol, 3.0 eq.) in DMF (2 mL) was added 5-(((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (20 mg, 0.071 mmol, 1.0 eq.) at room temperature. After warm stirring for 16 hours, H 2 O (10 mL) was added, and the mixture was extracted with DCM (10 mL*2). The organic layers were combined and concentrated in vacuo. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)pyridazine-3-carboxamide (15) (21 mg). MS: m/z=810 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.00(s,1H),9.08–9.03(m,1H),8.61(m,2H),8.26(m,2H),8.10(d,J＝8.9Hz,1H),7.83(d,J＝9.5Hz,1H),7.67(dd,J＝8.5,4.3Hz,1H),7.57(d,J＝8.7Hz,1H),7.42–7.27(m,3H),5.78–5.71(m,1H),4.71(m,1H),4.50(d,J＝13.2Hz,2H),4.14(d,J＝12.9Hz,2H),3.92(m,2H),3.65(m,2H),3.11(m,6H),2.91(s,2H),2.59(m,2H),2.23(m,3H),2.07(m,1H),2.01–1.92(m,2H),1.87(d,J＝12.4Hz,2H),1.70(m,3H),1.23(m,3H). 1 H NMR (400MHz, DMSO-d 6 )δ11.00(s,1H),9.08–9.03(m,1H),8.61(m,2H),8.26(m,2H),8.10(d,J＝8.9Hz,1H),7.83(d,J =9.5Hz,1 H),7.67(dd,J＝8.5,4.3Hz,1H),7.57(d,J＝8.7Hz,1H),7.42–7.27(m,3H),5.78–5.71(m,1H),4.71(m ,1H) ,4.50(d,J＝13.2Hz,2H),4.14(d,J＝12.9Hz,2H),3.92(m,2H),3.65(m,2H),3.11(m,6H),2.91(s, 2H),2.5 9(m,2H),2.23(m,3H),2.07(m,1H),2.01–1.92(m,2H),1.87(d,J＝12.4Hz,2H),1.70(m,3H),1.23 (m,3H).

实施例16化合物16的制备
Example 16 Preparation of Compound 16

第一步：化合物16-1的制备Step 1: Preparation of compound 16-1

室温下，向5-氟吡啶甲酸甲酯(1.0g，6.4mmol，1.0eq.)和哌啶-4-基甲醇(0.89g，7.7mmol，1.2eq.)的DMSO(3.5mL)溶液中加入K2CO3(1.1g，7.7mmol，1.2eq.)。在110℃搅拌1小时后，冷至室温，加入DCM(40mL)和H2O(20mL)。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩，得到5-(4-(羟甲基)哌啶-1-基)吡啶甲酸甲酯(16-1)(1.52g)。MS:m/z＝251[M+H]+ To a solution of methyl 5-fluoropicolinate (1.0 g, 6.4 mmol, 1.0 eq.) and piperidin-4-ylmethanol (0.89 g, 7.7 mmol, 1.2 eq.) in DMSO (3.5 mL) was added K 2 CO 3 (1.1 g, 7.7 mmol, 1.2 eq.) at room temperature. After stirring at 110°C for 1 hour, the mixture was cooled to room temperature and DCM (40 mL) and H 2 O (20 mL) were added. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give methyl 5-(4-(hydroxymethyl)piperidin-1-yl)picolinate (16-1) (1.52 g). MS: m/z=251[M+H] +

第二步：化合物16-2的制备Step 2: Preparation of compound 16-2

室温下，向5-(4-(羟甲基)哌啶-1-基)吡啶甲酸甲酯(16-1)(0.9g，3.6mmol，1.0eq.)的THF/H2O(10mL/10mL)溶液中加入LiOH·H2O(0.6克14.4毫摩尔，4.0eq.)。在35℃下搅拌16小时后，浓缩以除去THF，然后用1N HCl酸化至pH＝5。所得混合物通过反相色谱(HCOOH缓冲液)纯化，得到5-(4-(羟甲基)哌啶-1-基)皮考啉酸(16-2)(0.35g)。MS:m/z＝237[M+H]+ To a solution of 5-(4-(hydroxymethyl)piperidin-1-yl)picolinic acid methyl ester (16-1) (0.9 g, 3.6 mmol, 1.0 eq.) in THF/H 2 O (10 mL/10 mL) at room temperature was added LiOH·H 2 O (0.6 g 14.4 mmol, 4.0 eq.). After stirring at 35° C. for 16 hours, the mixture was concentrated to remove THF and then acidified to pH=5 with 1N HCl. The resulting mixture was purified by reverse phase chromatography (HCOOH buffer) to give 5-(4-(hydroxymethyl)piperidin-1-yl)picolinic acid (16-2) (0.35 g). MS: m/z=237 [M+H] +

第三步：化合物16-3的制备Step 3: Preparation of compound 16-3

室温下，向5-(4-(羟甲基)哌啶-1-基)吡啶甲酸(16-2)(120毫克，0.51毫摩尔，1.0eq.)和5-((((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(160毫克，0.54毫摩尔，1.05eq.)的DMF(6毫升)溶液中加入HBTU(208毫克，0.54毫摩尔，1.05eq.)和DIPEA(505uL，3.06毫摩尔，6.0eq.)。室温下搅拌16小时后，加入用H2O(80mL)，并用EA(80mL*3)萃取。合并有机层用饱和食盐水洗涤，减压浓缩。通过反相色谱(TFA缓冲液)纯化残留物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡啶酰胺(16-3)(100mg)。MS:m/z＝486[M+H]+ To a solution of 5-(4-(hydroxymethyl)piperidin-1-yl)picolinic acid (16-2) (120 mg, 0.51 mmol, 1.0 eq.) and 5-((((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (160 mg, 0.54 mmol, 1.05 eq.) in DMF (6 ml) at room temperature were added HBTU (208 mg, 0.54 mmol, 1.05 eq.) and DIPEA (505 uL, 3.06 mmol, 6.0 eq.). After stirring at room temperature for 16 hours, H 2 O (80 mL), and extracted with EA (80 mL * 3). The combined organic layers were washed with saturated brine and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-(hydroxymethyl)piperidin-1-yl)picolinamide (16-3) (100 mg). MS: m/z = 486 [M + H] +

第四步：化合物16-4的制备Step 4: Preparation of compound 16-4

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡啶酰胺(16-3)(92毫克，0.19毫摩尔，1.0eq.)的DCM(14毫升)溶液中加入Dess-Martin试剂(104毫克，0.25毫摩尔，1.3eq.)。在25℃下搅拌3小时后，加入饱和Na2S2O3水溶液(25mL)，并用DCM(25mL*3)萃取。合并有机层用饱和Na2S2O3水溶液(15mL)、饱和NaHCO3水溶液(15mL)和H2O(15mL)洗涤，用无水硫酸钠干燥，过滤减压浓缩，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡啶酰胺(16-4)(140mg)。MS:m/z＝484[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-(hydroxymethyl)piperidin-1-yl)picolinamide (16-3) (92 mg, 0.19 mmol, 1.0 eq.) in DCM (14 mL) was added Dess-Martin reagent (104 mg, 0.25 mmol, 1.3 eq.) at room temperature. After stirring at 25°C for 3 hours, saturated aqueous Na2S2O3 solution (25 mL ) was added, and the mixture was extracted with DCM (25 mL*3). The combined organic layers were washed with saturated aqueous Na2S2O3 solution ( 15 mL), saturated aqueous NaHCO3 solution (15 mL) and H2O (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-formylpiperidin-1-yl)picolinamide (16-4) (140 mg). MS: m/z=484 [M+H] +

第五步：化合物16的制备Step 5: Preparation of compound 16

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡啶酰胺(16-4)(130毫克，粗品，1.0eq.)和3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮(4-5)(90毫克，0.24毫摩尔，0.9eq.)的DCM(14毫升)溶液中加入TEA(13滴)。室温下搅拌2小时后，加入NaBH(OAc)3(143mg，0.67mmol，2.5eq.)。在室温下搅拌继续1小时，浓缩至干。向残留物中加入DCM/H2O(20mL+20mL)，并用DCM(20mL*2)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。通过反相色谱(TFA缓冲液)纯化残留物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1-氧代-1,2-二氢邻苯二甲酸-6-基)哌嗪-1-基)甲基)哌啶-1-基)吡啶甲酰胺(16)(31mg)。MS:m/z＝809[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-formylpiperidin-1-yl)picolinamide (16-4) (130 mg, crude, 1.0 eq.) and 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione (4-5) (90 mg, 0.24 mmol, 0.9 eq.) in DCM (14 mL) at room temperature was added TEA (13 drops). After stirring at room temperature for 2 hours, NaBH(OAc) 3 (143 mg, 0.67 mmol, 2.5 eq.) was added. Stirring was continued at room temperature for 1 hour and concentrated to dryness. DCM/H 2 O (20 mL+20 mL) was added to the residue and extracted with DCM (20 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalic acid-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)picolinamide (16) (31 mg). MS: m/z=809[M+H] +

1H NMR(400MHz,DMSO-d6)δ11.02(s,1H),9.54(br,1H),9.07(dd,J＝4.2,1.8Hz,1H),8.64(dd,J＝8.4,1.8Hz,1H),8.29(dd,J＝10.8,2.5Hz,3H),8.19(d,J＝8.2Hz,1H),8.12(d,J＝9.0Hz,1H),7.86(d,J＝8.8Hz,1H),7.69(dd,J＝8.4,4.2Hz,1H),7.60(dd,J＝9.0,1.4Hz,1H),7.44(dd,J＝8.8,2.5Hz,1H),7.38(d,J＝1.6Hz,1H),7.32(d,J＝8.4Hz,1H),5.77(dd,J＝12.0,5.4Hz,1H),4.79–4.66(br,1H),4.23–4.09(m,2H),3.97(d,J＝12.9Hz,2H),3.92–3.84(br,2H),3.33(t,J＝12.6Hz,2H),3.24–3.10(m,3H), 2.96–2.85(m,3H),2.64–2.57(m,1H),2.26–2.19(m,2H),2.15–2.05(m,2H),1.98–1.92(m,2H),1.90–1.82(m,2H),1.71–1.61(m,4H),1.35–1.27(m,2H),1.25–1.17(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ11.02(s,1H),9.54(br,1H),9.07(dd,J＝4.2,1.8Hz,1H),8.64(dd,J＝8.4,1.8Hz,1H),8.29(dd,J ＝10.8,2.5Hz,3H), 8.19(d,J＝8.2Hz,1H),8.12(d,J＝9.0Hz,1H),7.86(d,J＝8.8Hz,1H),7.69(dd,J＝8.4,4.2Hz,1H), 7.60(dd,J＝9.0,1.4H z,1H),7.44(dd,J＝8.8,2.5Hz,1H),7.38(d,J＝1.6Hz,1H),7.32(d,J＝8.4Hz,1H),5.77(dd,J＝12.0 ,5.4Hz,1H),4.79–4 .66(br,1H),4.23–4.09(m,2H),3.97(d,J＝12.9Hz,2H),3.92–3.84(br,2H),3.33(t,J＝12.6Hz,2H), 3.24–3.10(m,3H), 2.96–2.85(m,3H),2.64–2.57(m,1H),2.26–2.19(m,2H),2.15–2.05(m,2H),1.98–1.9 2(m,2H),1.90–1.82(m,2H),1.71–1.61(m,4H),1.35–1.27(m,2H),1.25–1.17(m,2H).

实施例17化合物17的制备
Example 17 Preparation of Compound 17

第一步：化合物17-1的制备Step 1: Preparation of compound 17-1

室温下，向5-氯吡嗪-2-羧酸甲酯(1.0g，5.8mmol，1.0eq.)和哌啶-4-基甲醇(0.89g，5.8mmol，1.0eq.)的DMSO(5mL)溶液中加入K2CO3(0.96g，6.9mmol，1.2eq.)。在110℃搅拌1小时后，冷至室温，加入DCM(40mL)和H2O(20mL)。有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并减压浓缩，得到5-(4-(羟甲基)哌啶-1-基)吡嗪-2-羧酸甲酯(17-1)(1.29克)。MS:m/z＝252[M+H]+ K 2 CO 3 (0.96 g, 6.9 mmol, 1.2 eq.) was added to a solution of 5-chloropyrazine-2-carboxylic acid methyl ester (1.0 g, 5.8 mmol, 1.0 eq.) and piperidin-4-ylmethanol (0.89 g, 5.8 mmol, 1.0 eq.) in DMSO (5 mL) at room temperature. After stirring at 110°C for 1 hour, the mixture was cooled to room temperature and DCM (40 mL) and H 2 O (20 mL) were added. The organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give 5-(4-(hydroxymethyl)piperidin-1-yl)pyrazine-2-carboxylic acid methyl ester (17-1) (1.29 g). MS: m/z=252[M+H] +

第二步：化合物17-2的制备Step 2: Preparation of compound 17-2

室温下，向5-(4-(羟甲基)哌啶-1-基)吡啶甲酸甲酯(17-1)(500mg，1.9mmol，1.0eq.)的THF/H2O(10mL/10mL)溶液中加入LiOH·H2O(316毫克7.9毫摩尔，4.0eq.)。在35℃下搅拌16小时后，浓缩以除去THF，然后用1N HCl酸化至pH＝5。所得混合物通过反相色谱(TFA缓冲液)纯化，得到5-(4-(羟甲基)哌啶-1-基)吡嗪-2-羧酸(17-2)(146mg)。MS:m/z＝238[M+H]+ To a solution of 5-(4-(hydroxymethyl)piperidin-1-yl)picolinic acid methyl ester (17-1) (500 mg, 1.9 mmol, 1.0 eq.) in THF/H 2 O (10 mL/10 mL) at room temperature was added LiOH·H 2 O (316 mg 7.9 mmol, 4.0 eq.). After stirring at 35° C. for 16 hours, the mixture was concentrated to remove THF and then acidified to pH=5 with 1N HCl. The resulting mixture was purified by reverse phase chromatography (TFA buffer) to give 5-(4-(hydroxymethyl)piperidin-1-yl)pyrazine-2-carboxylic acid (17-2) (146 mg). MS: m/z=238[M+H] +

第三步：化合物17-3的制备Step 3: Preparation of compound 17-3

室温下，向5-(4-(羟甲基)哌啶-1-基)吡嗪-2-羧酸(17-2)(120毫克，0.51毫摩尔，1.0eq.)和5-((((1r，4R)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(160毫克，0.54毫摩尔，1.05eq.)的DMF(6毫升)溶液中加入HBTU(208毫克，0.54毫摩尔，1.05eq.)和DIPEA(505uL，3.06毫摩尔，6.0eq.)。室温下搅拌16小时后，加入H2O(80mL)，并用EA(80mL*3)萃取。合并有机层用饱和食盐水洗涤并在减压下浓缩。残余物通过反相色谱(TFA缓冲液)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡嗪-2-甲酰胺(17-3)(100mg)。MS:m/z＝487[M+H]+ To a solution of 5-(4-(hydroxymethyl)piperidin-1-yl)pyrazine-2-carboxylic acid (17-2) (120 mg, 0.51 mmol, 1.0 eq.) and 5-((((1r,4R)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (160 mg, 0.54 mmol, 1.05 eq.) in DMF (6 mL) at room temperature were added HBTU (208 mg, 0.54 mmol, 1.05 eq.) and DIPEA (505 uL, 3.06 mmol, 6.0 eq.). After stirring at room temperature for 16 h, H 2 O (80mL), and extracted with EA (80mL*3). The combined organic layers were washed with saturated brine and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-(hydroxymethyl)piperidin-1-yl)pyrazine-2-carboxamide (17-3) (100 mg). MS: m/z=487[M+H] +

第四步：化合物17-4的制备Step 4: Preparation of compound 17-4

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-(羟甲基)哌啶-1-基)吡嗪-2-甲酰胺(17-3)(100毫克，0.2毫摩尔，1.0eq.)的DCM(3毫升)溶液中加入Dess-Martin试剂(104毫克，0.26毫摩尔，1.3eq.)。在25℃下搅拌3小时后，加入饱和Na2S2O3水溶液(25mL)中稀释，并用DCM(25mL*3)萃取。合并的有机层用饱和Na2S2O3水溶液(15mL)、饱和NaHCO3水溶液(15mL)和H2O(15mL)洗涤，用无水硫酸钠干燥，过滤减压浓缩，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺(17-4)(72mg)。MS:m/z＝485[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-(hydroxymethyl)piperidin-1-yl)pyrazine-2-carboxamide (17-3) (100 mg, 0.2 mmol, 1.0 eq.) in DCM (3 mL) was added Dess-Martin reagent (104 mg, 0.26 mmol, 1.3 eq.) at room temperature. After stirring at 25°C for 3 hours, the mixture was diluted with saturated aqueous Na2S2O3 solution (25 mL ) and extracted with DCM (25 mL*3). The combined organic layer was washed with saturated aqueous Na2S2O3 solution ( 15 mL), saturated aqueous NaHCO3 solution (15 mL) and H2O (15 mL), dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-formylpiperidin-1-yl)pyrazine-2-carboxamide (17-4) (72 mg). MS: m/z=485 [M+H] +

第五步：化合物17的制备Step 5: Preparation of compound 17

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-甲酰基哌啶-1-基)吡嗪-2-甲酰胺(17-4)(50毫克，粗品，1.0eq.)和3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮(4-5)(30毫克，0.09毫摩尔，0.9eq.)的DCM(4毫升)溶液中加入TEA(4滴)。室温下搅拌2h后，加入NaBH(OAc)3(63mg，0.3mmol，3.0eq.)。将所得混合物在室温下搅拌1小时，然后减压浓缩至干。向残留物中加入DCM/H2O(20mL+20mL)，并用DCM(20mL*2)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，减压浓缩。残余物通过反相色谱(TFA缓冲液)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-5-(4-((4-(2-(2,6-二氧代哌啶-3-基)-1-氧代-1,2-二氢邻苯二甲酸-6-基)哌嗪-1-基)甲基)哌啶-1-基)吡嗪-2-甲酰胺(17)(9.6mg)。MS:m/z＝810[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-formylpiperidin-1-yl)pyrazine-2-carboxamide (17-4) (50 mg, crude, 1.0 eq.) and 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione (4-5) (30 mg, 0.09 mmol, 0.9 eq.) in DCM (4 mL) was added TEA (4 drops) at room temperature. After stirring at room temperature for 2 h, NaBH(OAc) 3 (63 mg, 0.3 mmol, 3.0 eq.) was added. The resulting mixture was stirred at room temperature for 1 hour and then concentrated to dryness under reduced pressure. DCM/H 2 O (20 mL+20 mL) was added to the residue and extracted with DCM (20 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-5-(4-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalic acid-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)pyrazine-2-carboxamide (17) (9.6 mg). MS: m/z=810[M+H] +

1H NMR(400MHz,DMSO-d6)δ11.01(s,1H),9.58(s,1H),9.08(dd,J＝4.2,1.8Hz,1H),8.67–8.60(m,2H),8.30(s,1H),8.28(d,J＝2.8Hz,1H),8.16–8.08(m,2H),7.70(dd,J＝8.4,4.2Hz,1H),7.59(d,J＝8.8Hz,1H),7.37(s,1H),7.32(d,J＝8.4Hz,1H),5.79–5.74(m,1H),4.72(s,2H),4.51(d,J＝13.0Hz,2H),4.15(s,2H),3.91(s,1H),3.67(s,2H),3.07–3.01(m,2H),2.94–2.89(m,1H),2.68–2.65(m,1H),2.35–2.31(m,1H),2.24(s,2H),2.12–2.08(m,1H),1.98–1.86(m,4H),1.73–1.63(m,4H),1.30–1.15(m,4H). 1 H NMR (400MHz, DMSO-d 6 )δ11.01(s,1H),9.58(s,1H),9.08(dd,J＝4.2,1.8Hz,1H),8.67–8.60(m,2H),8.30(s,1H),8.28(d ,J＝2.8Hz,1H),8.16– 8.08(m,2H),7.70(dd,J＝8.4,4.2Hz,1H),7.59(d,J＝8.8Hz,1H),7.37(s,1H),7.32(d,J＝8.4Hz,1H ),5.79–5.74(m,1H),4 .72(s,2H),4.51(d,J＝13.0Hz,2H),4.15(s,2H),3.91(s,1H),3.67(s,2H),3.07–3.01(m,2H), 2.94–2.89(m,1H),2.68–2 .65(m,1H),2.35–2.31(m,1H),2.24(s,2H),2.12–2.08(m,1H),1.98–1.86(m,4H),1.73–1.63(m,4H) ,1.30–1.15(m,4H).

实施例18化合物18的制备
Example 18 Preparation of Compound 18

第一步：化合物18-1的制备Step 1: Preparation of compound 18-1

室温下，向2-(4-(羟甲基)哌啶-1-基(1.0g，4.1mmol，1.0eq.)和哌啶-4-基甲醇(668mg，5.8mmol，1.0eq.)的THF(10mL)溶液中加入DIEA(1.5g，11.6mmol，1.5eq.)。在室温下搅拌3小时，加入H2O(100mL)和DCM(100mL)，并用EA(3x 100mL)萃取。合并有机层用1N HCl(100mL)洗涤，用无水硫酸钠干燥，过滤浓缩，得到2-(4-(羟甲基)哌啶-1-基)嘧啶-5-羧酸乙酯(18-1)。MS:m/z＝265[M+H]+ To a solution of 2-(4-(hydroxymethyl)piperidin-1-yl (1.0 g, 4.1 mmol, 1.0 eq.) and piperidin-4-ylmethanol (668 mg, 5.8 mmol, 1.0 eq.) in THF (10 mL) was added DIEA (1.5 g, 11.6 mmol, 1.5 eq.) at room temperature. The mixture was stirred at room temperature for 3 hours, H 2 O (100 mL) and DCM (100 mL) were added, and the mixture was extracted with EA (3×100 mL). The combined organic layers were washed with 1N HCl (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to give ethyl 2-(4-(hydroxymethyl)piperidin-1-yl)pyrimidine-5-carboxylate (18-1). MS: m/z=265[M+H] +

第二步：化合物18-2的制备Step 2: Preparation of compound 18-2

室温下，向2-(4-(羟甲基)哌啶-1-基)嘧啶-5-羧酸乙酯(18-1)(1.1克，7.5毫摩尔，1.0eq.)的THF(10毫升)溶液中加入LiOH·H2O(945毫克，22.5毫摩尔，3.0eq.)的H2O(3mL)。在室温下搅拌 16小时后，加入2N HCl酸化至pH<4，并形成白色沉淀。然后将混合物过滤并干燥，得到2-(4-(羟甲基)哌啶-1-基)嘧啶-5-羧酸(18-2)(400毫克)。MS:m/z＝237[M+H]+ To a solution of ethyl 2-(4-(hydroxymethyl)piperidin-1-yl)pyrimidine-5-carboxylate (18-1) (1.1 g, 7.5 mmol, 1.0 eq.) in THF (10 mL) was added LiOH·H 2 O (945 mg, 22.5 mmol, 3.0 eq.) in H 2 O (3 mL) at room temperature. The mixture was stirred at room temperature. After 16 hours, 2N HCl was added to acidify to pH <4, and a white precipitate was formed. The mixture was then filtered and dried to give 2-(4-(hydroxymethyl)piperidin-1-yl)pyrimidine-5-carboxylic acid (18-2) (400 mg). MS: m/z = 237 [M+H] +

第三步：化合物18-3的制备Step 3: Preparation of compound 18-3

室温下，向2-(4-(羟甲基)哌啶-1-基)嘧啶-5-羧酸(18-2)(100mg，0.42mmol，1.0eq.)的DMF(15mL)溶液中加入DIEA(218mg，1.68mmol，4.0eq.)和HATU(160mg，0.42毫摩尔，1.0eq.)。室温下搅拌20分钟后，加入5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈盐酸盐(128毫克，0.42毫摩尔，1.0eq.)。将反应物在室温下搅拌2小时，加入H2O(20mL)和EA(20mL)，并用EA(3x 20mL)萃取水层。将有机层干燥并蒸发。残余物硅胶色谱(DCM/MeOH(20：1))纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-2-(4-(羟甲基)哌啶-1-基)嘧啶-5-甲酰胺(18-3)(50mg)。MS:m/z＝486[M+H]+ To a solution of 2-(4-(hydroxymethyl)piperidin-1-yl)pyrimidine-5-carboxylic acid (18-2) (100 mg, 0.42 mmol, 1.0 eq.) in DMF (15 mL) at room temperature were added DIEA (218 mg, 1.68 mmol, 4.0 eq.) and HATU (160 mg, 0.42 mmol, 1.0 eq.). After stirring at room temperature for 20 minutes, 5-(((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile hydrochloride (128 mg, 0.42 mmol, 1.0 eq.) was added. The reaction was stirred at room temperature for 2 hours, H 2 O (20 mL) and EA (20 mL) were added, and the aqueous layer was extracted with EA (3 x 20 mL). The organic layer was dried and evaporated. The residue was purified by silica gel chromatography (DCM/MeOH (20:1)) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-2-(4-(hydroxymethyl)piperidin-1-yl)pyrimidine-5-carboxamide (18-3) (50 mg). MS: m/z=486 [M+H] +

第四步：化合物18-4的制备Step 4: Preparation of compound 18-4

将N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-2-(4-(羟甲基)哌啶-1-基)嘧啶-5-甲酰胺(18-3)(50mg，0.102mmol，1.0eq.)和Dess-Martin试剂(49mg，0.113mmol，1.1eq.)的DCM(4mL)溶液在室温下搅拌12小时。加入H2O(50mL)和DCM(50mL)，并用DCM(3x50mL)萃取水层。将有机层干燥并蒸发。残余物通过反相色谱(TFA缓冲液)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-2-(4-甲酰基哌啶-1-基)嘧啶-5-甲酰胺(18-4)(20mg)。MS:m/z＝484[M+H]+ A solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-2-(4-(hydroxymethyl)piperidin-1-yl)pyrimidine-5-carboxamide (18-3) (50 mg, 0.102 mmol, 1.0 eq.) and Dess-Martin reagent (49 mg, 0.113 mmol, 1.1 eq.) in DCM (4 mL) was stirred at room temperature for 12 hours. H 2 O (50 mL) and DCM (50 mL) were added, and the aqueous layer was extracted with DCM (3×50 mL). The organic layer was dried and evaporated. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-2-(4-formylpiperidin-1-yl)pyrimidine-5-carboxamide (18-4) (20 mg). MS: m/z = 484 [M + H] +

第五步：化合物18的制备Step 5: Preparation of compound 18

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-2-(4-甲酰基哌啶-1-基)嘧啶-5-甲酰胺(18-4)(20毫克，0.04毫摩尔，1.0eq.)和3-(1-氧代-6-((哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮盐酸盐(4-5)(15.6毫克，0.04毫摩尔，1.0eq.)的DCM(2毫升)溶液中加入TEA(41毫克，0.41毫摩尔，10.0eq.)和MgSO4(49毫克，0.41毫摩尔，10.0eq.)。搅拌2小时后，加入NaBH(OAc)3(26mg，0.12mmol，3.0eq.)。在25℃下继续搅拌16小时，加入DCM(20毫升)和水(20毫升)。有机层浓缩。残余物经反相色谱(TFA缓冲液)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-2-(4-(4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哌啶-1-基)嘧啶-5-甲酰胺(18)(9mg)。MS:m/z＝810[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-2-(4-formylpiperidin-1-yl)pyrimidine-5-carboxamide (18-4) (20 mg, 0.04 mmol, 1.0 eq.) and 3-(1-oxo-6-((piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione hydrochloride (4-5) (15.6 mg, 0.04 mmol, 1.0 eq.) in DCM (2 ml) at room temperature were added TEA (41 mg, 0.41 mmol, 10.0 eq.) and MgSO 4 (49 mg, 0.41 mmol, 10.0 eq.). After stirring for 2 h, NaBH(OAc) 3 (26 mg, 0.12 mmol, 3.0 eq.). Stirring was continued at 25°C for 16 hours, and DCM (20 ml) and water (20 ml) were added. The organic layer was concentrated. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-2-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)pyrimidine-5-carboxamide (18) (9 mg). MS: m/z=810 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.02(s,1H),9.08(dd,J＝4.2,1.7Hz,1H),8.79(s,2H),8.64(dd,J＝8.5,1.8Hz,1H),8.35–8.26(m,2H),8.18(d,J＝7.4Hz,1H),8.12(d,J＝8.9Hz,1H),7.69(dd,J＝8.5,4.3Hz,1H),7.63–7.54(m,1H),7.43–7.32(m,2H),5.77(dd,J＝12.0,5.5Hz,1H),4.75(d,J＝12.5Hz,3H),4.16(d,J＝13.5Hz,2H),3.87(brs,1H),3.75–3.61(m,2H),3.38–3.27(m,2H),3.26–3.09(m,4H),3.08–2.98(m,2H),2.97–2.88(m,1H),2.68–2.55(m,2H),2.31–2.16(m,3H),2.15–2.06(m,1H),1.99(d,J＝11.5Hz,2H),1.88(d,J＝12.6Hz,2H),1.73–1.53(m,4H),1.26–1.13(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ11.02(s,1H),9.08(dd,J＝4.2,1.7Hz,1H),8.79(s,2H),8.64(dd,J＝8.5,1.8Hz,1H),8.35–8.26(m ,2H),8.18(d,J＝7.4Hz,1H),8.12(d, J＝8.9Hz,1H),7.69(dd,J＝8.5,4.3Hz,1H),7.63–7.54(m,1H),7.43–7.32(m,2H),5.77(dd,J＝12.0,5.5Hz ,1H),4.75(d,J＝12.5Hz,3H),4. 16(d,J＝13.5Hz,2H),3.87(brs,1H),3.75–3.61(m,2H),3.38–3.27(m,2H),3.26–3.09(m,4H),3.08–2.98( m,2H),2.97–2.88(m,1H),2.68 –2.55(m,2H),2.31–2.16(m,3H),2.15–2.06(m,1H),1.99(d,J＝11.5Hz,2H),1.88(d,J＝12.6Hz,2H), 1.73–1.53(m,4H),1.26–1.13(m,2H).

实施例19化合物19的制备
Example 19 Preparation of Compound 19

第一步：化合物19-1的制备Step 1: Preparation of compound 19-1

室温下，向6-氯烟酸甲酯(1.0g，5.8mmol，1.0eq.)和哌啶-4-甲醇(671mg，5.8mmol，1.0eq.)的DMSO(10mL)溶液中加入DIEA(2.2g，17.5mmol，3.0eq.)。在120℃搅拌3小时，冷至室温，加入H2O(50mL)和DCM(50mL)，并用EA(3x 50mL)萃取水层。合并有机层用饱和食盐水(100mL)洗涤，用无水硫酸钠干燥，过滤浓缩，得到1.2g 6-(4-(羟甲基)哌啶-1-基)烟酸甲酯(19-1)。MS:m/z＝251[M+H]+ To a solution of methyl 6-chloronicotinate (1.0 g, 5.8 mmol, 1.0 eq.) and piperidine-4-methanol (671 mg, 5.8 mmol, 1.0 eq.) in DMSO (10 mL) at room temperature was added DIEA (2.2 g, 17.5 mmol, 3.0 eq.). The mixture was stirred at 120°C for 3 hours, cooled to room temperature, H 2 O (50 mL) and DCM (50 mL) were added, and the aqueous layer was extracted with EA (3 x 50 mL). The combined organic layers were washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain 1.2 g of methyl 6-(4-(hydroxymethyl)piperidin-1-yl)nicotinate (19-1). MS: m/z=251[M+H] +

第二步：化合物19-2的制备Step 2: Preparation of compound 19-2

室温下，向6-(4-(羟甲基)哌啶-1-基)烟酸甲酯(19-1)(1.2g，4.8mmol，1.0eq.)的THF(10mL)溶液中加入LiOH·H2O(604毫克，14.3毫摩尔，3.0eq.)的H2O(3mL)溶液。室温下搅拌16小时后，减压浓缩。残余物用2N盐酸中和至pH＝7。残余物通过反相色谱(TFA缓冲液)纯化，得到6-(4-(羟甲基)哌啶-1-基)烟酸(19-2)(500mg)。MS:m/z＝237[M+H]+ To a solution of methyl 6-(4-(hydroxymethyl)piperidin-1-yl)nicotinate (19-1) (1.2 g, 4.8 mmol, 1.0 eq.) in THF (10 mL) was added a solution of LiOH·H 2 O (604 mg, 14.3 mmol, 3.0 eq.) in H 2 O (3 mL) at room temperature. After stirring at room temperature for 16 hours, the mixture was concentrated under reduced pressure. The residue was neutralized with 2N hydrochloric acid to pH = 7. The residue was purified by reverse phase chromatography (TFA buffer) to give 6-(4-(hydroxymethyl)piperidin-1-yl)nicotinic acid (19-2) (500 mg). MS: m/z = 237 [M+H] +

第三步：化合物19-3的制备Step 3: Preparation of compound 19-3

室温下，向6-(4-(羟甲基)哌啶-1-基)烟酸(19-2)(120mg，0.5mmol，1.0eq.)in DMF(15mL)中加入DIEA(258mg，2.0mmol，4.0eq.)和HBTU(285mg，0.75mmol，1.5eq.)。室温下搅拌20分钟后，加入5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈盐酸盐(1-7盐酸盐)(152毫克，0.5毫摩尔，1.0eq.)。在室温下搅拌2小时后，加入H2O(20mL)和EA(20mL)，并用EA(3x 20mL)萃取水层。将有机层干燥并蒸发。残余物用硅胶色谱(DCM/MeOH(20：1))纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)烟酰胺(19-3)(50mg)。MS:m/z＝486[M+H]+ To 6-(4-(hydroxymethyl)piperidin-1-yl)nicotinic acid (19-2) (120 mg, 0.5 mmol, 1.0 eq.) in DMF (15 mL) at room temperature were added DIEA (258 mg, 2.0 mmol, 4.0 eq.) and HBTU (285 mg, 0.75 mmol, 1.5 eq.). After stirring at room temperature for 20 minutes, 5-(((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile hydrochloride (1-7 hydrochloride) (152 mg, 0.5 mmol, 1.0 eq.) was added. After stirring at room temperature for 2 hours, H 2 O (20 mL) and EA (20 mL) were added, and the aqueous layer was extracted with EA (3 x 20 mL). The organic layer was dried and evaporated. The residue was purified by silica gel chromatography (DCM/MeOH (20:1)) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-(hydroxymethyl)piperidin-1-yl)nicotinamide (19-3) (50 mg). MS: m/z=486 [M+H] +

第四步：化合物19-4的制备Step 4: Preparation of compound 19-4

将N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-(羟甲基)哌啶-1-基)烟酰胺(19-3)(50mg，0.102mmol，1.0eq.)和Dess-Martin试剂(49mg，0.113mmol，1.1eq.)的DCM(4mL)溶液，室温下搅拌12小时。加入H2O(50mL)和DCM(50mL)稀释，并用DCM(3x 50mL)萃取水层。将有机层干燥并蒸发。残余物通过反相色谱(TFA缓冲液)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-甲酰基哌啶-1-基)烟酰胺(19-4)(30mg)。MS:m/z＝484[M+H]+ A solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-(hydroxymethyl)piperidin-1-yl)nicotinamide (19-3) (50 mg, 0.102 mmol, 1.0 eq.) and Dess-Martin reagent (49 mg, 0.113 mmol, 1.1 eq.) in DCM (4 mL) was stirred at room temperature for 12 hours. H 2 O (50 mL) and DCM (50 mL) were added to dilute, and the aqueous layer was extracted with DCM (3 x 50 mL). The organic layer was dried and evaporated. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-formylpiperidin-1-yl)nicotinamide (19-4) (30 mg). MS: m/z = 484 [M + H] +

第五步：化合物19的制备 Step 5: Preparation of compound 19

室温下，向N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-甲酰基哌啶-1-基)烟酰胺(19-4)(30毫克，0.06毫摩尔，1.0eq.)和3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮盐酸盐(4-5盐酸盐)(23毫克，0.06毫摩尔，1.0eq.)i的DCM(2mL)溶液中加入TEA(61毫克，0.6毫摩尔，10.0eq.)和MgSO4(73毫克，0.6毫摩尔，10.0eq.)。搅拌2小时后，加入NaBH(OAc)3(26mg，0.12mmol，3.0eq.)。继续搅拌16小时后，加入DCM(20毫升)和水(20毫升)。有机层浓缩。残余物经反相色谱(TFA缓冲液)纯化，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-(4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)哌啶-1-基)烟酰胺(19)(16.0mg)。MS:m/z＝809[M+H]+ To a solution of N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-formylpiperidin-1-yl)nicotinamide (19-4) (30 mg, 0.06 mmol, 1.0 eq.) and 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione hydrochloride (4-5 hydrochloride) (23 mg, 0.06 mmol, 1.0 eq.) in DCM (2 mL) at room temperature were added TEA (61 mg, 0.6 mmol, 10.0 eq.) and MgSO 4 (73 mg, 0.6 mmol, 10.0 eq.). After stirring for 2 hours, NaBH(OAc) 3 (26 mg, 0.12 mmol, 3.0 eq.) was added. After stirring for 16 hours, DCM (20 ml) and water (20 ml) were added. The organic layer was concentrated. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)piperazin-1-yl)methyl)piperidin-1-yl)nicotinamide (19) (16.0 mg). MS: m/z=809[M+H] +

1H NMR(400MHz,DMSO-d6)δ11.00(s,1H),9.05(dd,J＝4.3,1.8Hz,1H),8.65–8.54(m,2H),8.29–8.20(m,2H),8.08(dd,J＝9.5,2.6Hz,2H),7.96(dd,J＝9.0,2.5Hz,1H),7.67(dd,J＝8.5,4.3Hz,1H),7.57(d,J＝9.1Hz,1H),7.39–7.30(m,2H),6.87(d,J＝9.1Hz,1H),5.75(dd,J＝9.0,5.2Hz,1H),4.80–4.65(m,1H),4.48–4.42(m,1H),4.13–4.09(m,1H),3.90–3.78(m,1H),3.70–3.33(m,8H),3.20–3.11(m,3H),2.95–2.85(m,2H),2.65–2.50(m,1H),2.24–2.04(m,3H),2.24–2.20(m,4H),1.95–1.78(m,3H),1.65–1.50(m,4H),1.23–1.10(m,1H). 1 H NMR (400MHz, DMSO-d 6 )δ11.00(s,1H),9.05(dd,J＝4.3,1.8Hz,1H),8.65–8.54(m,2H),8.29–8.20(m,2H),8.08(dd,J＝9.5, 2.6Hz,2H),7.96(dd,J＝9.0 ,2.5Hz,1H),7.67(dd,J＝8.5,4.3Hz,1H),7.57(d,J＝9.1Hz,1H),7.39–7.30(m,2H),6.87(d,J＝9.1Hz ,1H),5.75(dd,J＝9.0,5.2H z,1H),4.80–4.65(m,1H),4.48–4.42(m,1H),4.13–4.09(m,1H),3.90–3.78(m,1H),3.70–3.33(m,8H), 3.20–3.11(m,3H),2.95– 2.85(m,2H),2.65–2.50(m,1H),2.24–2.04(m,3H),2.24–2.20(m,4H),1.95–1.78(m,3H),1.65–1.50(m,4H ),1.23–1.10(m,1H).

实施例20化合物20的制备
Example 20 Preparation of Compound 20

第一步：化合物20-1的制备Step 1: Preparation of compound 20-1

室温下，向叔丁基(1R，5S，6r)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-羧酸酯(100mg，0.47mmol，1.0eq.)的EA(0.5mL)溶液中加入HCl/EA(4M，0.5mL)。搅拌6小时后，浓缩，得到((1R，5S，6r)-3-氮杂双环[3.1.0]己烷-6-基)甲醇(20-1)(100毫克)。MS:m/z＝113[M+H]+ To a solution of tert-butyl (1R, 5S, 6r)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (100 mg, 0.47 mmol, 1.0 eq.) in EA (0.5 mL) was added HCl/EA (4 M, 0.5 mL) at room temperature. After stirring for 6 hours, the mixture was concentrated to give ((1R, 5S, 6r)-3-azabicyclo[3.1.0]hexane-6-yl)methanol (20-1) (100 mg). MS: m/z = 113 [M+H] +

第二步：化合物20-2的制备Step 2: Preparation of compound 20-2

室温下，向6-氯哒嗪-3-羧酸(142mg，0.89mmol，1.2eq.)，5-((((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(200mg，0.74mmol，1.0eq.)的DMF(4mL)溶液中加入HATU(313mg，1.0mmol，1.1eq.)和DIEA(400μL，2.67毫摩尔，3.0eq.)。搅拌6小时后，加入H2O(40mL)，并用EA(10mL *2)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在真空下浓缩。残余物在PE：EA＝3：1洗脱的硅胶上色谱纯化，得到6-氯-N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)哒嗪-3-甲酰胺(20-2)(250毫克)。MS:m/z＝407[M+H]+ To a solution of 6-chloropyridazine-3-carboxylic acid (142 mg, 0.89 mmol, 1.2 eq.), 5-((((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (200 mg, 0.74 mmol, 1.0 eq.) in DMF (4 mL) were added HATU (313 mg, 1.0 mmol, 1.1 eq.) and DIEA (400 μL, 2.67 mmol, 3.0 eq.) at room temperature. After stirring for 6 h, H 2 O (40 mL) was added and the mixture was washed with EA (10 mL) for 2 h. *2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum. The residue was chromatographed on silica gel eluted with PE:EA=3:1 to give 6-chloro-N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)pyridazine-3-carboxamide (20-2) (250 mg). MS: m/z=407[M+H] +

第三步：化合物20-3的制备Step 3: Preparation of compound 20-3

室温下，向6-氯-N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)哒嗪-3-甲酰胺(20-2)(50毫克，0.12毫摩尔，1.0eq.)和((1R，5S，6r)-3-氮杂双环[3.1.0]己烷-6-基)甲醇(20-1)(27毫克，0.24毫摩尔，2.0eq.)的1,4-dioxane(1mL)溶液中加入DIEA(32毫克，0.24毫摩尔，2.0eq.)。在100℃搅拌16小时后，冷至室温，浓缩，并用硅胶柱纯化(PE：EA＝1：1)，得到N-((1r，4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-((1R，5S，6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(20-3)(45mg)。MS:m/z＝484[M+H]+ To a solution of 6-chloro-N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)pyridazine-3-carboxamide (20-2) (50 mg, 0.12 mmol, 1.0 eq.) and ((1R,5S,6r)-3-azabicyclo[3.1.0]hexan-6-yl)methanol (20-1) (27 mg, 0.24 mmol, 2.0 eq.) in 1,4-dioxane (1 mL) was added DIEA (32 mg, 0.24 mmol, 2.0 eq.) at room temperature. After stirring at 100°C for 16 hours, the mixture was cooled to room temperature, concentrated, and purified by silica gel column (PE:EA=1:1) to give N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-((1R,5S,6R)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexane-3-yl)pyridazine-3-carboxamide (20-3) (45 mg). MS: m/z=484[M+H] +

第四步：化合物20-4的制备Step 4: Preparation of compound 20-4

将N-((1r，4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-((1R，5S，6R)-6-(羟甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(20-3)(40毫克，0.08毫摩尔，1.0eq.)，Dess-Martin高碘酸烷(40毫克，0.1毫摩尔，1.2eq.)的DCM(2毫升)溶液，室温下搅拌2小时。加入饱和Na2S2O3(10mL)稀释混合物，并分别用饱和NaHCO3(10mL)和H2O(10mL)洗涤，水层用DCM(10mL)萃取。将合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤，真空浓缩，得到N-((1r，4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-((1R，5S，6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(20-4)(40毫克)。MS:m/z＝482[M+H]+ A solution of N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-((1R,5S,6R)-6-(hydroxymethyl)-3-azabicyclo[3.1.0]hexan-3-yl)pyridazine-3-carboxamide (20-3) (40 mg, 0.08 mmol, 1.0 eq.) and Dess-Martin periodate (40 mg, 0.1 mmol, 1.2 eq.) in DCM (2 mL) was stirred at room temperature for 2 hours. Saturated Na 2 S 2 O 3 (10 mL) was added to dilute the mixture, and the mixture was washed with saturated NaHCO 3 (10 mL) and H 2 O (10 mL), respectively, and the aqueous layer was extracted with DCM (10 mL). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated in vacuo to give N-((1r, 4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-((1R, 5S, 6R)-6-formyl-3-azabicyclo[3.1.0]hexane-3-yl)pyridazine-3-carboxamide (20-4) (40 mg). MS: m/z=482[M+H] +

第五步：化合物20的制备Step 5: Preparation of compound 20

向N-((1r，4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(((1R，5S，6R)-6-甲酰基-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(20-4)(100毫克，0.21毫摩尔，1.0eq.)，3-(1-氧代-6-(哌嗪-1-基)酞嗪-2(1H)-基)哌啶-2，6-二酮(4-5)(70毫克，0.21毫摩尔，1.0eq.)的DCM(4毫升)溶液中加入TEA(0.1毫升)和MgSO4(50毫克)，将反应混合物室温下搅拌16小时，然后加入NaBH(OAc)3(53mg，0.25mmol，1.2eq.)。加入H2O(10mL)稀释混合物，并用DCM(10mL*2)萃取。合并有机层，真空浓缩。残余物通过反相色谱(TFA缓冲液)纯化得到N-((1r，4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-((1R，5S，6S)-6-((4-(2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)哌嗪-1-基)甲基)-3-氮杂双环[3.1.0]己烷-3-基)哒嗪-3-甲酰胺(20)(11毫克)。MS:m/z＝808[M+H]+ To a solution of N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(((1R,5S,6R)-6-formyl-3-azabicyclo[3.1.0]hexan-3-yl)pyridazine-3-carboxamide (20-4) (100 mg, 0.21 mmol, 1.0 eq.), 3-(1-oxo-6-(piperazin-1-yl)phthalazin-2(1H)-yl)piperidine-2,6-dione (4-5) (70 mg, 0.21 mmol, 1.0 eq.) in DCM (4 ml) were added TEA (0.1 ml) and MgSO 4 (50 mg), the reaction mixture was stirred at room temperature for 16 h, then NaBH(OAc) 3 (53 mg, 0.25 mmol, 1.2 eq.) was added. H 2 O was added. O (10mL) dilute the mixture and extract with DCM (10mL * 2). Combine the organic layers and concentrate in vacuo. The residue is purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-((1R, 5S, 6S)-6-((4-(2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazine-6-yl)piperazine-1-yl)methyl)-3-azabicyclo[3.1.0]hexane-3-yl)pyridazine-3-carboxamide (20) (11 mg). MS: m/z=808[M+H] +

1H NMR(400MHz,DMSO-d6)δ11.00(s,1H),9.08–9.02(m,1H),8.64–8.55(m,2H),8.26(m,2H),8.09(d,J＝9.0Hz,1H),7.84(d,J＝9.4Hz,1H),7.67(dd,J＝8.5,4.3Hz,1H),7.57(d,J＝9.0Hz,1H),7.38–7.28(m,2H),6.97(d,J＝9.4Hz,1H),5.74(m,1H),4.71(m,1H),4.17(m,2H),3.90(m,2H),3.67(m,2H),3.55(m,2H),3.24–3.11(m,4H),2.88(m,2H),2.59(m,3H),2.22(m,2H),2.07(m,2H),1.92(m,3H),1.70(m,3H),1.21(m,2H),1.05(m,1H). 1 H NMR (400MHz, DMSO-d 6 )δ11.00(s,1H),9.08–9.02(m,1H),8.64–8.55(m,2H),8.26(m,2H),8.09(d,J＝9.0Hz,1H),7.84(d ,J＝ 9.4Hz,1H),7.67(dd,J＝8.5,4.3Hz,1H),7.57(d,J＝9.0Hz,1H),7.38–7.28(m,2H),6.97(d,J＝9.4Hz, 1H ),5.74(m,1H),4.71(m,1H),4.17(m,2H),3.90(m,2H),3.67(m,2H),3.55(m,2H),3.24–3.11(m, 4H),2. 88(m,2H),2.59(m,3H),2.22(m,2H),2.07(m,2H),1.92(m,3H),1.70(m,3H),1.21(m,2H),1.05 (m,1H).

实施例21化合物21的制备
Example 21 Preparation of Compound 21

室温下,向3-(1-甲基-7-(((R)-吡咯烷-3-基甲氧基)-1H-吲唑-3-基)哌啶-2，6-二酮(21-6)(30毫克，0.09毫摩尔，1.0eq.)，N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(42毫克，0.09毫摩尔，1.0eq.)的DCM(2毫升)溶液中加入TEA(0.5毫升)。搅拌16小时后，加入NaBH(OAc)3(23毫克，0.10毫摩尔，1.2eq.)。反应完成后，浓缩并通过反相柱纯化，MeCN：H2O＝55％，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(3R)-3-((3-(2，6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(21)(25毫克)。MS:m/z＝809[M+H]+ To a solution of 3-(1-methyl-7-(((R)-pyrrolidin-3-ylmethoxy)-1H-indazol-3-yl)piperidine-2,6-dione (21-6) (30 mg, 0.09 mmol, 1.0 eq.), N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (42 mg, 0.09 mmol, 1.0 eq.) in DCM (2 ml) was added TEA (0.5 ml) at room temperature. After stirring for 16 h, NaBH(OAc) 3 (23 mg, 0.10 mmol, 1.2 eq.) was added. After completion of the reaction, it was concentrated and purified by reverse phase column, MeCN:H 2 O = 55%, to obtain N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(3R)-3-((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (21) (25 mg). MS: m/z = 809 [M+H] +

1H NMR(400MHz,DMSO-d6)δ10.87(s,1H),9.04(dd,J＝4.2,1.7Hz,1H),8.61(dd,J＝8.6,1.8Hz,1H),8.26(d,J＝8.3Hz,1H),7.99(d,J＝7.6Hz,1H),7.73(d,J＝8.5Hz,2H),7.66(dd,J＝8.5,4.3Hz,1H),7.32(d,J＝8.4Hz,1H),7.23(d,J＝8.1Hz,1H),7.04-6.90(m,3H),6.83(d,J＝7.2Hz,1H),4.76-4.64(m,2H),4.31(dd,J＝9.7,5.1Hz,1H),4.16(s,3H),3.92-3.78(m,4H),3.74-3.66(m,1H),3.20-3.10(m,4H),2.76(t,J＝12.1Hz,2H),2.70–2.53(m,3H),2.40–2.10(m,5H),2.04-1.90(m,4H),1.88-1.76(m,3H),1.70-1.50(m,4H),1.34-1.19(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ10.87(s,1H),9.04(dd,J＝4.2,1.7Hz,1H),8.61(dd,J＝8.6,1.8Hz,1H),8.26(d,J＝8.3Hz,1H), 7.99(d,J＝7.6Hz,1H),7.73(d, J＝8.5Hz,2H),7.66(dd,J＝8.5,4.3Hz,1H),7.32(d,J＝8.4Hz,1H),7.23(d,J＝8.1Hz,1H),7.04-6.90( m,3H),6.83(d,J=7.2Hz,1H) ,4.76-4.64(m,2H),4.31(dd,J＝9.7,5.1Hz,1H),4.16(s,3H),3.92-3.78(m,4H),3.74-3.66(m,1H),3.20 -3.10(m,4H),2.76(t,J ＝12.1Hz,2H),2.70–2.53(m,3H),2.40–2.10(m,5H),2.04-1.90(m,4H),1.88-1.76(m,3H),1.70-1.50(m,4H ),1.34-1.19(m,2H).

实施例22化合物22的制备
Example 22 Preparation of Compound 22

第一步：化合物22-1的制备Step 1: Preparation of compound 22-1

室温下,向6-溴酞嗪-1(2H)-酮(50毫克，0.22毫摩尔，1.0eq.)和B2Pin2(68毫克，0.26毫摩尔，1.2eq.)的二恶烷(2毫升)溶液中加入Pd(dppf)Cl2·CH2Cl2(15毫克，0.02毫摩尔，0.08eq.)和KOAc(44毫克，0.44毫摩尔，2.0eq.)。在100℃下搅拌16小时后，冷至室温，浓缩。残余物通过反相柱纯化，得到6-(4，4，5，5-四甲基-1，3，2-二氧杂硼烷-2-基)酞嗪-1(2H)-酮(22-1)(60mg)。MS:m/z＝273[M+H]+ Pd(dppf)Cl 2 ·CH 2 Cl 2 (15 mg, 0.02 mmol, 0.08 eq.) and KOAc (44 mg, 0.44 mmol, 2.0 eq.) were added to a solution of 6-bromophthalazin-1(2H)-one (50 mg, 0.22 mmol, 1.0 eq.) and B 2 Pin 2 ( 68 mg , 0.26 mmol, 1.2 eq.) in dioxane (2 ml) at room temperature. After stirring at 100° C. for 16 hours, the mixture was cooled to room temperature and concentrated. The residue was purified by reverse phase column to give 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phthalazin-1(2H)-one (22-1) (60 mg). MS: m/z=273[M+H] +

第二步：化合物22-2的制备Step 2: Preparation of compound 22-2

室温下,向6-(4，4，5，5-四甲基-1，3，2-二氧杂硼烷-2-基)酞嗪-1(2H)-酮(22-1)(60毫克，0.22毫摩尔，1.0eq.)的THF(3毫升)溶液中加入30％H2O2(125μL，1.1毫摩尔，5.0eq.)。室温搅拌16小时后，浓缩，得到6-羟基酞嗪-1(2H)-酮(22-2)(60mg)，用于下一步而不进行纯化。MS:m/z＝163[M+H]+ To a solution of 6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phthalazin-1(2H)-one (22-1) (60 mg, 0.22 mmol, 1.0 eq.) in THF (3 ml) was added 30% H 2 O 2 (125 μL, 1.1 mmol, 5.0 eq.) at room temperature. After stirring at room temperature for 16 hours, the mixture was concentrated to give 6-hydroxyphthalazin-1(2H)-one (22-2) (60 mg), which was used in the next step without purification. MS: m/z=163[M+H] +

第三步：化合物22-3的制备Step 3: Preparation of compound 22-3

室温下,向6-羟基酞嗪-1(2H)-酮(22-2)(60毫克，0.22毫摩尔，1.0eq.)和叔丁基(S)-3-((对甲苯磺酰氧基)甲基)吡咯烷-1-羧酸叔丁酯(118毫克，0.33毫摩尔，1.5eq.)的DMF(2毫升)溶液中加入K2CO3(76毫克，0.55毫摩尔，2.5eq.)。在80℃搅拌16小时后，冷至室温，通过反相色谱(TFA缓冲液)纯化反应混合物，得到(S)-3-(((1-氧代-1，2-二氢邻苯二甲嗪-6-基)氧基)甲基)吡咯烷-1-羧酸酯(22-3)(40mg)。MS:m/z＝346[M+H]+ K 2 CO 3 (76 mg, 0.55 mmol, 2.5 eq.) was added to a solution of 6-hydroxyphthalazin- 1 (2H)-one (22-2) (60 mg, 0.22 mmol, 1.0 eq.) and tert-butyl (S)-3-((toluenesulfonyloxy)methyl)pyrrolidine-1-carboxylate (118 mg, 0.33 mmol, 1.5 eq.) in DMF (2 ml) at room temperature. After stirring at 80° C. for 16 hours, the mixture was cooled to room temperature and the reaction mixture was purified by reverse phase chromatography (TFA buffer) to give (S)-3-(((1-oxo-1,2-dihydrophthalazin-6-yl)oxy)methyl)pyrrolidine-1-carboxylate (22-3) (40 mg). MS: m/z=346 [M+H] +

第四步：化合物22-4的制备Step 4: Preparation of compound 22-4

室温下,向(S)-3-(((1-氧代-1，2-二氢邻苯二甲嗪-6-基)氧基)甲基)吡咯烷-1-羧酸叔丁酯(22-3)(40mg，0.12mmol，1.0eq.)的DMF溶液中加入NaH(60％重量，10毫克，0.36毫摩尔，3.0eq.)。在40℃下搅拌1小时后，加入3-溴哌啶-2，6-二酮(34mg，0.18mmol，1.5eq.)在0.3mL DMF中的溶液，并在40℃下搅拌16小时。冷至室温，加入饱和NH4Cl水溶液(10mL)，并用EA(10mL*3)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。残余物通过反相色谱(TFA缓冲液)纯化，得到叔丁基(3S)-3-(((2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)氧基)甲基)吡咯烷-1-羧酸酯(22-4)(52mg)。MS:m/z＝457[M+H]+ To a solution of (S)-tert-butyl 3-(((1-oxo-1,2-dihydrophthalazin-6-yl)oxy)methyl)pyrrolidine-1-carboxylate (22-3) (40 mg, 0.12 mmol, 1.0 eq.) in DMF was added NaH (60% weight, 10 mg, 0.36 mmol, 3.0 eq.) at room temperature. After stirring at 40°C for 1 hour, a solution of 3-bromopiperidine-2,6-dione (34 mg, 0.18 mmol, 1.5 eq.) in 0.3 mL of DMF was added and stirred at 40°C for 16 hours. After cooling to room temperature, a saturated aqueous NH 4 Cl solution (10 mL) was added and extracted with EA (10 mL*3). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give tert-butyl (3S)-3-(((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)oxy)methyl)pyrrolidine-1-carboxylate (22-4) (52 mg). MS: m/z=457 [M+H] +

第五步：化合物22-5的制备Step 5: Preparation of compound 22-5

室温下,向叔丁基(3S)-3-((2-(2，6-二氧代哌啶-3-基)-1-氧代-1，2-二氢邻苯二甲嗪-6-基)氧基)甲基)吡咯烷-1-羧酸酯(22-4)(52毫克，0.12毫摩尔，1.0eq.)在EA(1毫升)中的溶液中加入HCl(5mL,4M in EA)。搅拌16小时后，浓缩，得到3-(1-氧代-6-((S)-吡咯烷-3-基甲氧基)酞嗪-2(1H)-基)哌啶-2，6-二酮盐酸盐(22-5)(35毫克)。MS:m/z＝357[M+H]+ To a solution of tert-butyl (3S)-3-((2-(2,6-dioxopiperidin-3-yl)-1-oxo-1,2-dihydrophthalazin-6-yl)oxy)methyl)pyrrolidine-1-carboxylate (22-4) (52 mg, 0.12 mmol, 1.0 eq.) in EA (1 ml) was added HCl (5 mL, 4 M in EA) at room temperature. After stirring for 16 hours, the mixture was concentrated to give 3-(1-oxo-6-((S)-pyrrolidin-3-ylmethoxy)phthalazin-2(1H)-yl)piperidine-2,6-dione hydrochloride (22-5) (35 mg). MS: m/z=357[M+H] +

第六步：化合物22的制备Step 6: Preparation of compound 22

室温下,向3-(1-氧代-6-(((S)-吡咯烷-3-甲氧基)酞嗪-2(1H)-基)哌啶-2，6-二酮盐酸盐(22-5)(35毫克，0.12毫摩尔，1.0eq.)和N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(43毫克，0.12毫摩尔，1.0eq.)的DCM(7mL)中加入TEA(4滴)和MgSO4(107毫克，1.2毫摩尔，10.0eq.)。搅拌16小时后，加入NaBH(OAc)3(48毫克，0.30毫摩尔，2.5eq.)。在室温下再搅拌1小时后，浓缩至干。加入EA/H2O(20mL+20mL)，并用EA(20mL*2)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在减压下浓缩。通过反相色谱(TFA缓冲液)纯化残留物，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3S)-3-((2-(2,6-二氧代阿片苷-3-基)-1-氧代-1,2-二氢邻苯二甲酸-6-基)氧)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(22)(9mg)。MS:m/z＝823.0[M+H]+ To a solution of 3-(1-oxo-6-(((S)-pyrrolidin-3-methoxy)phthalazin-2(1H)-yl)piperidine-2,6-dione hydrochloride (22-5) (35 mg, 0.12 mmol, 1.0 eq.) and N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (43 mg, 0.12 mmol, 1.0 eq.) in DCM (7 mL) was added TEA (4 drops) and MgSO 4 (107 mg, 1.2 mmol, 10.0 eq.) at room temperature. After stirring for 16 h, NaBH(OAc) 3 (48 mg, 0.30 mmol, 2.5 eq.) was added. After stirring for another 1 h at room temperature, the mixture was concentrated to dryness. EA/H 2 O (20mL + 20mL), and extracted with EA (20mL * 2). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r, 4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3S)-3-((2-(2,6-dioxoopioside-3-yl)-1-oxo-1,2-dihydrophthalic acid-6-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (22) (9 mg). MS: m / z = 823.0 [M + H] +

实施例23化合物23的制备
Example 23 Preparation of Compound 23

23-1的合成：
Synthesis of 23-1:

向2-氟-3-硝基苯甲腈(23-1-1)(8.60g，51.8mmol，1.0eq.)于DMF(86mL)中的溶液中添加甲基肼(22.4g，155mmol，3.0eq.)和DIEA(50.2g，388mmol，7.5eq.)。将反应物在25℃下搅拌1小时。TLC(PE/EA 1:1)显示反应完成。添加EA(800mL)和水(850mL)。将有机层浓缩至100mL。将混合物冷却至室温并过滤。干燥残余物，得到呈红色固体状的1-甲基-7-硝基-1H-吲唑-3-胺(23-1-2)(7.71g，77％产率)。To a solution of 2-fluoro-3-nitrobenzonitrile (23-1-1) (8.60 g, 51.8 mmol, 1.0 eq.) in DMF (86 mL) was added methylhydrazine (22.4 g, 155 mmol, 3.0 eq.) and DIEA (50.2 g, 388 mmol, 7.5 eq.). The reactants were stirred at 25 °C for 1 hour. TLC (PE/EA 1:1) showed that the reaction was complete. EA (800 mL) and water (850 mL) were added. The organic layer was concentrated to 100 mL. The mixture was cooled to room temperature and filtered. The residue was dried to give 1-methyl-7-nitro-1H-indazole-3-amine (23-1-2) (7.71 g, 77% yield) as a red solid.

将乳酸(2.15g，85％，20.3mmol，1.3eq.)和DBU(3.09g，20.3mmol，1.3eq.)的混合物在25℃下搅拌16小时。添加1-甲基-7-硝基-1H-吲唑-3-胺(23-1-2)(3.00g，15.6mmol，1.0eq.)和丙烯酸甲酯(9.40g，109mmol，7.0eq.)。将反应物在85℃下搅拌40小时。TLC(PE/EA 1:1)显示反应未完成。添加EA(100mL)和水(100mL)。浓缩有机层。将残余物用柱(PE/EA 3:1)纯化，得到呈红色固体状的3-((1-甲基-7-硝基-1H-吲唑-3-基)氨基)丙酸甲酯(23-1-3)(2.08g，54％产率)。A mixture of lactic acid (2.15 g, 85%, 20.3 mmol, 1.3 eq.) and DBU (3.09 g, 20.3 mmol, 1.3 eq.) was stirred at 25 °C for 16 h. 1-Methyl-7-nitro-1H-indazol-3-amine (23-1-2) (3.00 g, 15.6 mmol, 1.0 eq.) and methyl acrylate (9.40 g, 109 mmol, 7.0 eq.) were added. The reaction was stirred at 85 °C for 40 h. TLC (PE/EA 1:1) showed that the reaction was not completed. EA (100 mL) and water (100 mL) were added. The organic layer was concentrated. The residue was purified by column (PE/EA 3:1) to give methyl 3-((1-methyl-7-nitro-1H-indazol-3-yl)amino)propanoate (23-1-3) (2.08 g, 54% yield) as a red solid.

向3-((1-甲基-7-硝基-1H-吲唑-3-基)氨基)丙酸甲酯(23-1-3)(1.98g，7.13mmol，1.0eq.)于AcOH(20mL)中的溶液中添加KCNO(1.73g，21.4mmol，3.0eq.)。将反应物在25℃下搅拌16小时。TLC(纯EA)显示反应几乎完成。添加水(100mL)和EA(100mL)。浓缩有机层。将残余物用柱(PE/EA 1:1至纯EA)纯化，得到呈黄色固体状的3-(1-(1-甲基-7-硝基-1H-吲唑-3-基)脲基)丙酸甲酯(23-1-4)(1.26g，61％产率)。To a solution of methyl 3-((1-methyl-7-nitro-1H-indazol-3-yl)amino)propanoate (23-1-3) (1.98 g, 7.13 mmol, 1.0 eq.) in AcOH (20 mL) was added KCNO (1.73 g, 21.4 mmol, 3.0 eq.). The reactants were stirred at 25 °C for 16 hours. TLC (pure EA) showed that the reaction was almost complete. Water (100 mL) and EA (100 mL) were added. The organic layer was concentrated. The residue was purified by column (PE/EA 1:1 to pure EA) to give methyl 3-(1-(1-methyl-7-nitro-1H-indazol-3-yl)ureido)propanoate (23-1-4) (1.26 g, 61% yield) as a yellow solid.

向3-(1-(1-甲基-7-硝基-1H-吲唑-3-基)脲基)丙酸甲酯(23-1-4)(1.26g，3.92mmol，1.0eq.)于MeCN(13mL)中的溶液中添加Triton-B(492mg，40％于水中，1.18mmol，0.3eq.)。将反应物在15℃下搅拌72小时。TLC显示反应完成。添加水(100mL)和EA(100mL)。浓缩有机层，得到呈黄色固体状的1-(1-甲基-7-硝基-1H-吲唑-3-基)二氢嘧啶-2,4(1H,3H)-二酮(23-1-5)(1.04g，92％产率)。Triton-B (492 mg, 40% in water, 1.18 mmol, 0.3 eq.) was added to a solution of methyl 3-(1-(1-methyl-7-nitro-1H-indazol-3-yl)ureido)propanoate (23-1-4) (1.26 g, 3.92 mmol, 1.0 eq.) in MeCN (13 mL). The reactant was stirred at 15 ° C for 72 hours. TLC showed that the reaction was complete. Water (100 mL) and EA (100 mL) were added. The organic layer was concentrated to give 1-(1-methyl-7-nitro-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (23-1-5) (1.04 g, 92% yield) as a yellow solid.

向1-(1-甲基-7-硝基-1H-吲唑-3-基)二氢嘧啶-2,4(1H,3H)-二酮(23-1-5)(1.00g，3.11mmol，1.0eq.)于MeOH(10mL)和THF(10mL)中的溶液中添加Fe(520mg，9.34mmol，1.0eq.)和NH4Cl水溶液(10％，10mL)。将反应物在70℃下搅拌4小时。TLC显示反应完成。过滤混合物用THF(50mL)洗涤滤饼。浓缩滤液。添加水(100mL)和EA(100mL)。用EA(5×100mL)萃取水层。浓缩合并的有机层，得到呈红色固体状的1-(7-氨基-1-甲基-1H-吲唑-3-基)二氢嘧啶-2,4(1H,3H)-二酮(23-1)(820mg，92％产率)。To a solution of 1-(1-methyl-7-nitro-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (23-1-5) (1.00 g, 3.11 mmol, 1.0 eq.) in MeOH (10 mL) and THF (10 mL) was added Fe (520 mg, 9.34 mmol, 1.0 eq.) and NH 4 Cl aqueous solution (10%, 10 mL). The reactants were stirred at 70° C. for 4 hours. TLC showed that the reaction was complete. The mixture was filtered and the filter cake was washed with THF (50 mL). The filtrate was concentrated. Water (100 mL) and EA (100 mL) were added. The aqueous layer was extracted with EA (5×100 mL). The combined organic layers were concentrated to give 1-(7-amino-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (23-1) (820 mg, 92% yield) as a red solid.

23-3的合成：
Synthesis of 23-3:

室温下，向叔丁基(1R，5S，6r)-6-((苄氧基)甲基)-3-氮杂双环[3.1.0]己烷-3-羧酸酯(2-1)(1.2g，3.7mmol，1.0eq.)的THF/H2O/MeOH(8mL/2mL/2mL)溶液中加入LiOH·H2O(333毫克，9.3毫摩尔，2.5当量)。将反应混合物在40℃搅拌16小时后，用H2O(30mL)和EA(15mL)稀释混合物，并在减压下浓缩。将所得混合物用1N HCl中和至pH＝5，并用EA(50mL*3)萃取。合并有机层，饱和食盐水洗涤，无水硫酸钠干燥，过滤，减压浓缩，得到4-(4-(叔丁氧羰基)哌嗪-1-基)苯甲酸(23-3-1)(1.03g)。MS:m/z＝307[M+H]+ To a solution of tert-butyl (1R, 5S, 6r)-6-((benzyloxy)methyl)-3-azabicyclo[3.1.0]hexane-3-carboxylate (2-1) (1.2 g, 3.7 mmol, 1.0 eq.) in THF/H 2 O/MeOH (8 mL/2 mL/2 mL) was added LiOH·H 2 O (333 mg, 9.3 mmol, 2.5 eq.) at room temperature. The reaction mixture was stirred at 40°C for 16 hours, then diluted with H 2 O (30 mL) and EA (15 mL), and concentrated under reduced pressure. The resulting mixture was neutralized to pH=5 with 1N HCl, and extracted with EA (50 mL*3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)benzoic acid (23-3-1) (1.03 g). MS: m/z = 307 [M + H] +

室温下，向4-(4-(叔丁氧羰基)哌嗪-1-基)苯甲酸(23-3-1)(120mg，0.39mmol，1.0当量)的DMF(5mL)溶液中加入HBTU(180g，46.8mmol，1.2当量)和DIEA(435微升，2.34毫摩尔，6.0当量)。在30℃搅拌1小时后，然后加入5-(((1r，4r)-4-氨基环己氧基)喹啉-8-甲腈盐酸盐(1-7)(230mg，0.74mmol，1.9当量)。再将反应混合物在30℃下搅拌3小时后，用H2O(50mL)稀释，并用EA(50mL*3)萃取。合并有机层，饱和食盐水洗涤，无水硫酸钠干燥，过滤，减压浓缩。残余物通过反相色谱(TFA缓冲液)纯化，得到4-(4-(1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)氨基甲酰基)苯基)哌嗪-1-羧酸叔丁酯(23-3-2)(210mg)。MS:m/z＝556[M+H]+ To a solution of 4-(4-(tert-butoxycarbonyl)piperazin-1-yl)benzoic acid (23-3-1) (120 mg, 0.39 mmol, 1.0 eq) in DMF (5 mL) at room temperature were added HBTU (180 g, 46.8 mmol, 1.2 eq) and DIEA (435 μL, 2.34 mmol, 6.0 eq). After stirring at 30°C for 1 hour, 5-(((1r,4r)-4-aminocyclohexyloxy)quinoline-8-carbonitrile hydrochloride (1-7) (230 mg, 0.74 mmol, 1.9 equivalents) was added. After stirring the reaction mixture at 30°C for 3 hours, it was diluted with H 2 O (50 mL) and extracted with EA (50 mL*3). The organic layers were combined, washed with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by reverse phase chromatography (TFA buffer) to give tert-butyl 4-(4-(1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)carbamoyl)phenyl)piperazine-1-carboxylate (23-3-2) (210 mg). MS: m/z=556[M+H] +

室温下，向4-(4-(1r，4r)-4-((8-氰基喹啉-5-氧基)环己基)氨基甲酰基)苯基)哌嗪-1-羧酸叔丁酯(23-3-2)(210毫克，0.38毫摩尔，1.0当量)的EA(2毫升)溶液中加入HCl(5毫升，4M in EA)。室温下搅拌16小时后，浓缩得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(哌嗪-1-基)苯甲酰胺(23-3)(205毫克)。MS:m/z＝456[M+H]+ To a solution of tert-butyl 4-(4-(1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)carbamoyl)phenyl)piperazine-1-carboxylate (23-3-2) (210 mg, 0.38 mmol, 1.0 eq) in EA (2 ml) was added HCl (5 ml, 4 M in EA) at room temperature. After stirring at room temperature for 16 hours, the mixture was concentrated to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(piperazin-1-yl)benzamide (23-3) (205 mg). MS: m/z=456[M+H] +

合物23的制备Preparation of Compound 23

室温下,向1-(7-氨基-1-甲基-1H-吲唑-3-基)二氢嘧啶-2，4(1H，3H)-二酮(23-1)(50mg、0.19mmol、1.0eq.)在DCM(1mL)和H2O(0.5mL)的溶液中，依次加入溴乙酰氯(18μL、0.21mmol、1.1eq.)、K2CO3(30mg、0.21mmol、1.1eq.)。搅拌6小时后，加入H2O(20mL)，并用EA(5mL*2)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在真空下浓缩，得到2-溴-N-(3-(2，4-二氧代四氢嘧啶-1(2H)-基)-1-甲基-1H-吲唑-7-基)乙酰胺(23-2)(80毫克)。MS:m/z＝379[M+H]+ To a solution of 1-(7-amino-1-methyl-1H-indazol-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (23-1) (50 mg, 0.19 mmol, 1.0 eq.) in DCM (1 mL) and H 2 O (0.5 mL) at room temperature, bromoacetyl chloride (18 μL, 0.21 mmol, 1.1 eq.) and K 2 CO 3 (30 mg, 0.21 mmol, 1.1 eq.) were added in sequence. After stirring for 6 hours, H 2 O (20 mL) was added and extracted with EA (5 mL*2). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to give 2-bromo-N-(3-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-1-methyl-1H-indazol-7-yl)acetamide (23-2) (80 mg). MS: m/z = 379 [M + H] +

第二步：化合物23的制备Step 2: Preparation of compound 23

室温下,向2-溴-N-(3-(2，4-二氧代四氢嘧啶-1(2H)-基)-1-甲基-1H-吲唑-7-基)乙酰胺(23-2)(55毫克，0.15毫摩尔，1.5eq.)，N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(哌嗪 -1-基)苯甲酰胺23-3(48毫克，0.1毫摩尔，1.0eq.)的DMF(1mL)溶液中加入K2CO3(28毫克，0.2毫摩尔，2.0eq.)。在80℃搅拌2小时，冷至室温，浓缩，HPLC纯化(MeCN：H2O＝55％)，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-((2-(3-(2，4-二氧代四氢嘧啶-1(2H)-基)-1-甲基-1H-吲唑-7-基)氨基)-2-氧代乙基)哌嗪-1-基)苯甲酰胺(23)(18mg)。MS:m/z＝755[M+H]+ At room temperature, 2-bromo-N-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-7-yl)acetamide (23-2) (55 mg, 0.15 mmol, 1.5 eq.), N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(piperazine To a solution of 4-((1r,4r) -4 -((8-cyanoquinolin-5-yl) oxy )cyclohexyl)-4-(4-((2-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-7-yl)amino)-2-oxoethyl)piperazin-1-yl)benzamide (23-3) (48 mg, 0.1 mmol, 1.0 eq.) in DMF (1 mL) was added K 2 CO 3 (28 mg, 0.2 mmol, 2.0 eq.). The mixture was stirred at 80°C for 2 hours, cooled to room temperature, concentrated, and purified by HPLC (MeCN:H 2 O=55%) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-((2-(3-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-1-methyl-1H-indazol-7-yl)amino)-2-oxoethyl)piperazin-1-yl)benzamide (23) (18 mg). MS: m/z=755[M+H] +

1H NMR(400MHz,DMSO-d6)δ10.56(s,1H),9.04(dd,J＝4.2,1.7Hz,1H),8.61(dd,J＝8.5,1.7Hz,1H),8.26(d,J＝8.3Hz,1H),8.04(d,J＝7.6Hz,1H),7.76(d,J＝8.4Hz,2H),7.66(dd,J＝8.5,4.2Hz,1H),7.52(d,J＝8.2Hz,1H),7.32(d,J＝8.4Hz,1H),7.27(d,J＝7.3Hz,1H),7.09(t,J＝7.7Hz,1H),6.98(d,J＝8.5Hz,2H),4.76-4.64(m,1H),4.08(s,3H),3.92–3.80(m,3H),3.48-3.18(m,8H),2.75(t,J＝6.7Hz,2H),2.28–2.18(m,2H),1.96–1.88(m,2H),1.70–1.51(m,4H),1.26–1.19(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ10.56(s,1H),9.04(dd,J＝4.2,1.7Hz,1H),8.61(dd,J＝8.5,1.7Hz,1H),8.26(d,J＝8.3Hz,1H), 8.04(d,J＝7.6Hz ,1H),7.76(d,J＝8.4Hz,2H),7.66(dd,J＝8.5,4.2Hz,1H),7.52(d,J＝8.2Hz,1H),7.32(d,J＝8.4Hz ,1H),7.27(d,J＝ 7.3Hz,1H),7.09(t,J＝7.7Hz,1H),6.98(d,J＝8.5Hz,2H),4.76-4.64(m,1H),4.08(s,3H),3.92–3.80( m,3H),3.48 -3.18(m,8H),2.75(t,J＝6.7Hz,2H),2.28–2.18(m,2H),1.96–1.88(m,2H),1.70–1.51(m,4H),1.26–1.19 (m,2H).

实施例24化合物24的制备
Example 24 Preparation of Compound 24

第一步：化合物24-1的制备Step 1: Preparation of compound 24-1

室温下,向6-(4-甲酰基哌啶-1-基)哒嗪-3-羧酸叔丁酯(15-3)(300mg，1.03mmol，1.0eq.)的DCM(3mL)溶液中加入2-(2，6-二氧代哌啶-3-基)-5-(((R)-吡咯烷-3-甲氧基)异吲哚啉-1，3-二酮盐酸盐(5-4)(405mg，1.03毫摩尔，1.0eq.)和TEA(1.04克，10.3毫摩尔，10.0eq.)。搅拌2小时hou1，加入NaBH(OAc)3(436毫克，2.06毫摩尔，2.0eq.)。室温下继续搅拌16h后，浓缩。残余物用反相色谱(水(0.05％ TFA)和MeCN)纯化，得到6-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哒啶-1-基)哒嗪-3-羧酸酯(24-1)(170mg)。MS:m/z＝632[M+H]+ To a solution of tert-butyl 6-(4-formylpiperidin-1-yl)pyridazine-3-carboxylate (15-3) (300 mg, 1.03 mmol, 1.0 eq.) in DCM (3 mL) at room temperature were added 2-(2,6-dioxopiperidin-3-yl)-5-(((R)-pyrrolidine-3-methoxy)isoindoline-1,3-dione hydrochloride (5-4) (405 mg, 1.03 mmol, 1.0 eq.) and TEA (1.04 g, 10.3 mmol, 10.0 eq.). The mixture was stirred for 2 h and then NaBH(OAc) 3 (436 mg, 2.06 mmol, 2.0 eq.) was added. The mixture was stirred for 16 h at room temperature and then concentrated. The residue was purified by reverse phase chromatography (water (0.05% TFA) and MeCN) to give 6-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)pyridin-1-yl)pyridazine-3-carboxylate (24-1) (170 mg). MS: m/z=632[M+H] +

第二步：化合物24-2的制备Step 2: Preparation of compound 24-2

室温下,向6-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哒嗪-3-羧酸叔丁酯(24-1)(85毫克，0.147毫摩尔，1.0eq.)的DCM(2毫升)溶液中加入TFA(1毫升)。室温下搅拌2小时后，浓缩得到6-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哒啶-1-基)哒嗪-3-羧酸(24-2)(70毫克)。MS:m/z＝576[M+H]+ To a solution of tert-butyl 6-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)pyridazine-3-carboxylate (24-1) (85 mg, 0.147 mmol, 1.0 eq.) in DCM (2 ml) was added TFA (1 ml) at room temperature. After stirring at room temperature for 2 hours, the mixture was concentrated to give 6-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)pyrididin-1-yl)pyridazine-3-carboxylic acid (24-2) (70 mg). MS: m/z=576[M+H] +

第三步：化合物24的制备Step 3: Preparation of compound 24

室温下,向6-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基) 甲基)吡咯烷-1-基)甲基)哒啶-1-基)哒嗪-3-羧酸(24-2)(70毫克，0.12毫摩尔，1.0eq.)和5-(1r，4R)-4-氨基环己氧基)喹啉-8-甲腈(1-7)(33毫克，0.12毫摩尔，1.0eq.)的DMF(2毫升)溶液中加入DIEA(31毫克，0.24毫摩尔，2.0eq.)和HATU(55毫克)，0.14毫摩尔，1.2eq.)。搅拌2h后，反相色谱法纯化混合物，用水(0.05％TFA)和MeCN洗脱得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哒啶-1-基)哒嗪-3-甲酰胺(24)(29.4mg)。MS:m/z＝826[M+H]+ At room temperature, 6-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy) To a solution of 5-(1R,4R)-4-aminocyclohexyloxy)quinoline-8-carbonitrile (1-7) (33 mg, 0.12 mmol, 1.0 eq.) in DMF (2 ml) were added DIEA (31 mg, 0.24 mmol, 2.0 eq.) and HATU (55 mg, 0.14 mmol, 1.2 eq.). After stirring for 2 h, the mixture was purified by reverse phase chromatography, eluting with water (0.05% TFA) and MeCN to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)pyridin-1-yl)pyridazine-3-carboxamide (24) (29.4 mg). MS: m/z=826 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.09(s,1H),9.08–9.02(m,1H),8.60(dd,J＝10.5,8.2Hz,2H),8.26(d,J＝8.2Hz,1H),7.84(dd,J＝16.9,8.9Hz,2H),7.67(dd,J＝8.4,4.3Hz,1H),7.47(dd,J＝5.2,2.2Hz,1H),7.36(dd,J＝8.2,2.5Hz,2H),7.32(d,J＝8.3Hz,1H),5.10(dd,J＝12.8,5.4Hz,1H),4.71(brs,1H),4.49(d,J＝13.2Hz,2H),4.31–4.13(m,2H),3.97–4.82(m,1H),3.75–3.63(m,1H),3.21–3.11(m,3H),3.08–2.94(m,3H),2.91–2.78(m,3H),2.62–2.49(m,2H),2.27–2.18(m,2H),2.17–2.00(m,3H),1.97–1.78(m,5H),1.74–1.58(m,4H),1.28–1.14(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ11.09(s,1H),9.08–9.02(m,1H),8.60(dd,J＝10.5,8.2Hz,2H),8.26(d,J＝8.2Hz,1H),7.84(dd,J ＝16.9,8.9Hz,2H),7.67(dd,J＝8 .4,4.3Hz,1H),7.47(dd,J＝5.2,2.2Hz,1H),7.36(dd,J＝8.2,2.5Hz,2H),7.32(d,J＝8.3Hz,1H),5.10 (dd,J＝12.8,5.4Hz,1H),4.71( brs,1H),4.49(d,J＝13.2Hz,2H),4.31–4.13(m,2H),3.97–4.82(m,1H),3.75–3.63(m,1H),3.21–3.11(m, 3H),3.08–2.94(m,3H),2. 91–2.78(m,3H),2.62–2.49(m,2H),2.27–2.18(m,2H),2.17–2.00(m,3H),1.97–1.78(m,5H),1.74–1.58(m ,4H),1.28–1.14(m,2H).

实施例25化合物25的制备
Example 25 Preparation of Compound 25

第一步：化合物25-1的制备Step 1: Preparation of compound 25-1

室温下,向叔丁基((1s，4s)-4-羟基环己基)氨基甲酸酯(1g，3.46mmol，1.0eq.)，4-甲基苯磺酰氯(1g，5.2mmol，1.5eq.)在CHCl3(8mL)中的溶液中加入吡啶(3mL)。在80℃下搅拌16小时后，冷至室温，加入H2O(50mL)，并用DCM(20mL*2)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，真空过滤浓缩，用硅胶色谱纯化(PE：EA＝2：1)，得到(1s，4s)-4-((叔丁氧羰基)氨基)环己基-4-甲基苯磺酸酯(25-1)(1g)。Pyridine ( 3 mL) was added to a solution of tert-butyl ((1s, 4s)-4-hydroxycyclohexyl)carbamate (1 g, 3.46 mmol, 1.0 eq.) and 4-methylbenzenesulfonyl chloride (1 g, 5.2 mmol, 1.5 eq.) in CHCl 3 (8 mL) at room temperature. After stirring at 80°C for 16 hours, the mixture was cooled to room temperature, H 2 O (50 mL) was added, and the mixture was extracted with DCM (20 mL*2). The combined organic layer was washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by silica gel chromatography (PE:EA=2:1) to give (1s, 4s)-4-((tert-butyloxycarbonyl)amino)cyclohexyl-4-methylbenzenesulfonate (25-1) (1 g).

MS:m/z＝391[M+H+23]+ MS: m/z = 391 [M + H + 23] +

第二步：化合物25-2的制备Step 2: Preparation of compound 25-2

将5.8-二溴喹恶啉(800毫克，2.78毫摩尔，1.0eq.)，CuI(529毫克，2.78毫摩尔，1.0eq.)，NaOMe(165毫克，3.06毫摩尔，1.1eq.)在甲醇(0.5毫升)和DMF(20毫升)中的反应混合物在N2气氛下于100℃搅拌16小时。冷至室温，加入H2O(20mL)，并用EA(50mL*2)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，真空过滤浓缩，硅胶色谱纯化，PE：EA＝1：1洗脱，得到5-溴-8-甲 氧基喹恶啉(25-2)(300mg)。MS:m/z＝239[M+H]+ The reaction mixture of 5,8-dibromoquinoxaline (800 mg, 2.78 mmol, 1.0 eq.), CuI (529 mg, 2.78 mmol, 1.0 eq.), NaOMe (165 mg, 3.06 mmol, 1.1 eq.) in methanol (0.5 ml) and DMF (20 ml) was stirred at 100°C for 16 hours under N2 atmosphere. After cooling to room temperature, H2O (20 mL) was added and extracted with EA (50 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by silica gel chromatography with PE:EA=1:1 as eluent to obtain 5-bromo-8-methyl Oxyquinoxaline (25-2) (300 mg). MS: m/z = 239 [M + H] +

第三步：化合物25-3的制备Step 3: Preparation of compound 25-3

将5-溴-8-甲氧基喹恶啉(25-2)(200mg，0.69毫摩尔，1.0eq.)，CuCN(200毫克，2.08毫摩尔，3.0eq.)的DMF(5mL)溶液在N2气氛下在100℃搅拌16小时。冷至室温，加入H2O(10mL)，并用EA(10mL*2)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，过滤并在真空下浓缩，用MeCN：H2O＝95％色谱纯化，得到8-甲氧基喹恶啉-5-甲腈(25-3)(60mg)。MS:m/z＝185[M+H]+ A solution of 5-bromo-8-methoxyquinoxaline (25-2) (200 mg, 0.69 mmol, 1.0 eq.) and CuCN (200 mg, 2.08 mmol, 3.0 eq.) in DMF (5 mL) was stirred at 100°C for 16 hours under N2 atmosphere. After cooling to room temperature, H2O (10 mL) was added and extracted with EA (10 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, filtered and concentrated under vacuum, and purified by chromatography using MeCN:H2O=95% to obtain 8-methoxyquinoxaline-5-carbonitrile (25-3) (60 mg). MS: m/z=185[M+H] +

第四步：化合物25-4的制备Step 4: Preparation of compound 25-4

室温下,向8-甲氧基喹恶啉-5-甲腈(25-3)(45mg，0.05mmol，1.0eq.)的DCM(1mL)溶液中缓慢加入BBr3(1M in DCM)(0.8mL，0.3mmol，6.0eq.)中，然后在40℃下搅拌反应混合物1小时。冷至室温，加入用H2O(10mL)淬灭混合物，并用DCM(10mL*3)萃取。将组合的有机层在真空下浓缩，得到黄色固体(70mg，粗)的8-羟基喹喔啉-5-甲腈(25-4)(70mg)。MS:m/z＝171[M+H]+ At room temperature, BBr 3 (1M in DCM) (0.8 mL, 0.3 mmol, 6.0 eq.) was slowly added to a solution of 8-methoxyquinoxaline-5-carbonitrile (25-3) (45 mg, 0.05 mmol, 1.0 eq.) in DCM (1 mL), and the reaction mixture was stirred at 40° C. for 1 hour. After cooling to room temperature, the mixture was quenched with H 2 O (10 mL) and extracted with DCM (10 mL*3). The combined organic layers were concentrated under vacuum to give 8-hydroxyquinoxaline-5-carbonitrile (25-4) (70 mg) as a yellow solid (70 mg, crude). MS: m/z=171[M+H] +

第五步：化合物25-5的制备Step 5: Preparation of compound 25-5

室温下,向8-羟基喹恶啉-5-甲腈(25-4)(70毫克，0.41毫摩尔，1.0eq.)，(1s，4s)-4-((叔丁氧羰基)氨基)环己基-4-甲基苯磺酸酯(25-1)(227毫克，0.61毫摩尔，1.5eq.)的DMF(2毫升)溶液中加入K2CO3(115毫克，0.82毫摩尔，2.0eq.)。在80℃搅拌16小时后，冷至室温，加入H2O(20mL)，并用EA(20mL*2)萃取。合并的有机层用饱和食盐水洗涤，用无水硫酸钠干燥，真空过滤浓缩，HPLC纯化(MeCN：H2O＝80％)，得到叔丁基((1r，4r)-4-((8-氰基喹喔啉-5-基)氧基)环己基)氨基甲酸酯(25-5)(11mg)。MS:m/z＝368[M+H]+ K 2 CO 3 (115 mg, 0.82 mmol, 2.0 eq.) was added to a DMF (2 mL) solution of 8-hydroxyquinoxaline-5-carbonitrile (25-4) (70 mg, 0.41 mmol, 1.0 eq.), (1s, 4s)-4-((tert-butyloxycarbonyl)amino ) cyclohexyl-4-methylbenzenesulfonate (25-1) (227 mg, 0.61 mmol, 1.5 eq.) at room temperature. After stirring at 80° C. for 16 hours, the mixture was cooled to room temperature, H 2 O (20 mL) was added, and the mixture was extracted with EA (20 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by HPLC (MeCN: H2O =80%) to obtain tert-butyl ((1r,4r)-4-((8-cyanoquinoxalin-5-yl)oxy)cyclohexyl)carbamate (25-5) (11 mg). MS: m/z=368 [M+H] +

第六步：化合物25-6的制备Step 6: Preparation of compound 25-6

室温下,向叔丁基(((1r，4r)-4-((8-氰基喹喔啉-5-基)氧基)环己基)氨基甲酸酯(25-5)(11mg，0.03mmol，1.0eq.)在EA(1mL)中的溶液中加入4M HCl/EA(0.5mL)。搅拌16小时后，浓缩，得到8-(((1r，4r)-4-氨基环己基)氧基)喹恶啉-5-甲腈(25-6)(10毫克)。MS:m/z＝268[M+H]+ To a solution of tert-butyl (((1r, 4r)-4-((8-cyanoquinoxalin-5-yl)oxy)cyclohexyl)carbamate (25-5) (11 mg, 0.03 mmol, 1.0 eq.) in EA (1 mL) was added 4M HCl/EA (0.5 mL) at room temperature. After stirring for 16 hours, the mixture was concentrated to give 8-(((1r, 4r)-4-aminocyclohexyl)oxy)quinoxaline-5-carbonitrile (25-6) (10 mg). MS: m/z=268 [M+H] +

第七步：化合物25的制备Step 7: Preparation of compound 25

室温下,向8-(((1r，4r)-4-氨基环己氧基)喹恶啉-5-甲腈(25-6)(10毫克，0.03毫摩尔，1.0eq.)的溶液中，加入6-(4-(3R)-3-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哒啶-1-基)哒嗪-3-羧酸(24-2)(26毫克，0.04毫摩尔，1.2eq.)，HATU(17毫克，0.04毫摩尔，1.2eq.)的DMF(1mL)溶液中加入DIEA(25μL，0.1毫摩尔，3.0eq.)。搅拌4小时后，HPLC纯化(用MeCN：H2O＝55％)，得到N-((1r，4r)-4-((8-氰基喹恶啉-5-基)氧基)环己基)-6-(4-(3R)-3-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哒啶-3-甲酰胺(25)(10毫克)。MS:m/z＝827[M+H]+ To a solution of 8-(((1r,4r)-4-aminocyclohexyloxy)quinoxaline-5-carbonitrile (25-6) (10 mg, 0.03 mmol, 1.0 eq.), 6-(4-(3R)-3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)pyridin-1-yl)pyridazine-3-carboxylic acid (24-2) (26 mg, 0.04 mmol, 1.2 eq.), HATU (17 mg, 0.04 mmol, 1.2 eq.) in DMF (1 mL) was added DIEA (25 μL, 0.1 mmol, 3.0 eq.) at room temperature. After stirring for 4 hours, the mixture was purified by HPLC (using MeCN: H2O =55%) to give N-((1r,4r)-4-((8-cyanoquinoxalin-5-yl)oxy)cyclohexyl)-6-(4-(3R)-3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)pyrididine-3-carboxamide (25) (10 mg). MS: m/z=827 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.17(s,1H),9.18(d,J＝6.2Hz,1H),9.12(d,J＝6.1Hz,1H),8.71–8.63(m,1H),8.48–8.41(m,1H),7.93–7.85(m,2H),7.66–7.58(m,1H),7.54–7.49(m,1H),7.47–7.39(m,2H),5.43–5.35(m,1H),5.24–5.11(m,2H),4.57–4.50(m,2H),4.21–4.14(m,2H),3.13–3.03(m,4H),2.98–2.91(m,2H),2.35–2.25(m,4H),2.13–1.99(m,8H),1.94–1.87(m,2H),1.78–1.71(m, 4H),1.58–1.49(m,4H). 1 H NMR (400MHz, DMSO-d6) δ11.17(s,1H),9.18(d,J＝6.2Hz,1H),9.12(d,J＝6.1Hz,1H),8.71–8.63( m,1H),8.48–8.41(m,1H),7.93–7.85(m,2H),7.66–7.58(m,1H),7.54–7.49(m,1H),7.47–7.39(m ,2H),5.43–5.35(m,1H),5.24–5.11(m,2H),4.57–4.50(m,2H),4.21–4.14(m,2H),3.13–3.03(m ,4H),2.98–2.91(m,2H),2.35–2.25(m,4H),2.13–1.99(m,8H),1.94–1.87(m,2H),1.78–1.71(m, 4H),1.58–1.49(m,4H).

实施例26化合物26的制备
Example 26 Preparation of Compound 26

第一步：化合物26-1的制备Step 1: Preparation of compound 26-1

室温下,向2-甲基异烟酸甲酯(1mL，6.6mmol，1.0eq.)的MeOH(10mL)溶液中加入4M HCl的EA(5mL)。搅拌1小时后，浓缩以得到白色固体。将白色固体加入MeOH(20毫升)和PtO2(100毫克)，将反应混合物在H2气氛(50Psi)下室温搅拌6小时，过滤，滤饼在真空下浓缩，得到顺-2-甲基哌啶-4-羧酸甲酯(26-1)(1.0克)。MS:m/z＝157[M+H]+ To a solution of methyl 2-methylisonicotinate (1 mL, 6.6 mmol, 1.0 eq.) in MeOH (10 mL) at room temperature was added 4 M HCl in EA (5 mL). After stirring for 1 hour, the mixture was concentrated to give a white solid. The white solid was added to MeOH (20 mL) and PtO 2 (100 mg), and the reaction mixture was stirred at room temperature for 6 hours under H 2 atmosphere (50 Psi), filtered, and the filter cake was concentrated under vacuum to give cis-2-methylpiperidine-4-carboxylic acid methyl ester (26-1) (1.0 g). MS: m/z=157 [M+H] +

第二步：化合物26-2的制备Step 2: Preparation of compound 26-2

室温下,向顺-2-甲基哌啶-4-羧酸甲酯(26-1)(1g，6.37毫摩尔，1.0eq.)THF(20mL)溶液中缓慢加入LiAlH4(1M in THF)(9.5mL，9.55mmol，1.5eq.)。搅拌16小时后，依次加入H2O(0.3mL)、15％NaOH(0.3mL)和H2O(0.9mL)淬灭混合物。将反应混合物过滤并在真空下浓缩滤液，得到顺-2-甲基哌啶-4-基甲醇(26-2)(1克)。MS:m/z＝129[M+H]+ At room temperature, LiAlH 4 (1M in THF) (9.5 mL, 9.55 mmol, 1.5 eq.) was slowly added to a solution of cis-2-methylpiperidine-4-carboxylic acid methyl ester (26-1) (1 g, 6.37 mmol, 1.0 eq.) in THF (20 mL). After stirring for 16 hours, H 2 O (0.3 mL), 15% NaOH (0.3 mL) and H 2 O (0.9 mL) were added in sequence to quench the mixture. The reaction mixture was filtered and the filtrate was concentrated under vacuum to give cis-2-methylpiperidin-4-ylmethanol (26-2) (1 g). MS: m/z=129[M+H] +

第三步：化合物26-3的制备Step 3: Preparation of compound 26-3

室温下,向6-氯哒嗪-3-羧酸叔丁酯(500毫克，2.33毫摩尔，1.0eq.)，((2R，4R)-2-甲基哌啶-4-基)甲醇(26-2)(600毫克，4.66毫摩尔，2.0eq.)的溶液中加入DIEA(1毫升，4.66毫摩尔，2.0eq.)。在100℃下搅拌16小时后，冷至室温，浓缩，并在硅胶柱纯化(PE：EA＝1：1)，得到6-(顺-4-(羟甲基)-2-甲基哌啶-1-基)哒嗪-3-羧酸叔丁酯(26-3)(110mg)。MS:m/z＝307[M+H]+ At room temperature, DIEA (1 ml, 4.66 mmol, 2.0 eq.) was added to a solution of tert-butyl 6-chloropyridazine-3-carboxylate (500 mg, 2.33 mmol, 1.0 eq.) and ((2R, 4R)-2-methylpiperidin-4-yl)methanol (26-2) (600 mg, 4.66 mmol, 2.0 eq.). After stirring at 100°C for 16 hours, the mixture was cooled to room temperature, concentrated, and purified on a silica gel column (PE:EA=1:1) to give tert-butyl 6-(cis-4-(hydroxymethyl)-2-methylpiperidin-1-yl)pyridazine-3-carboxylate (26-3) (110 mg). MS: m/z=307[M+H] +

第四步：化合物26-4的制备Step 4: Preparation of compound 26-4

室温下,将6-(顺-4-(羟甲基)-2-甲基哌啶-1-基)哒嗪-3-羧酸叔丁酯(26-3)(110mg，0.36mmol，1.0eq.)，Dess-Martin高碘酸烷(180mg，0.43mmol，1.2eq.)在DCM(2mL)中的混合物以室温搅拌2小时。加入DCM(10mL)稀释，并分别用饱和Na2S2O3(10mL)，饱和NaHCO3(10mL)和H2O(10mL)洗涤。将有机层在真空下浓缩，得到6-(顺-4-甲酰基-2-甲基哌啶-1-基)哒嗪-3-羧酸叔丁酯(26-4)(100mg)。MS:m/z＝305[M+H]+ At room temperature, a mixture of 6-(cis-4-(hydroxymethyl)-2-methylpiperidin-1-yl)pyridazine-3-carboxylic acid tert-butyl ester (26-3) (110 mg, 0.36 mmol, 1.0 eq.), Dess-Martin periodic acid alkane (180 mg, 0.43 mmol, 1.2 eq.) in DCM (2 mL) was stirred at room temperature for 2 hours. DCM (10 mL) was added for dilution, and washed with saturated Na 2 S 2 O 3 (10 mL), saturated NaHCO 3 (10 mL) and H 2 O (10 mL), respectively. The organic layer was concentrated under vacuum to give 6-(cis-4-formyl-2-methylpiperidin-1-yl)pyridazine-3-carboxylic acid tert-butyl ester (26-4) (100 mg). MS: m/z=305[M+H] +

第五步：化合物26-5的制备Step 5: Preparation of compound 26-5

室温下,将6-(顺-4-甲基哌啶-1-基)哒嗪-3-羧酸叔丁酯(26-4)(100mg，0.32mmol，1.0eq.)，2-(2，6-二氧代哌啶-3-基)-5-(((R)-吡咯烷-3-基甲氧基)异吲哚啉-1，3-二酮(5-4)(140mg，0.39毫摩尔，1.2eq.)和TEA(200μL，0.96毫摩尔，3.0eq.)在DCM(2mL)溶液中的反应混合物室温搅拌2小时，然后将反应混合物加入NaBH(OAc)3(85毫克，0.39毫摩尔，1.2eq.)，搅拌16小时后，加H2O(20mL)稀释，并用DCM(10mL*2)萃取。合并有机层用饱和食盐水洗涤，用无水硫酸钠干燥，真空过滤浓缩，用MeCN：H2O＝60％色谱纯化，得到6-(顺-4-(((3R)-3-(((2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)哒嗪-3-羧酸叔丁酯(26-5)(100mg)。MS:m/z＝646[M+H]+ At room temperature, a reaction mixture of tert-butyl 6-(cis-4-methylpiperidin-1-yl)pyridazine-3-carboxylate (26-4) (100 mg, 0.32 mmol, 1.0 eq.), 2-(2,6-dioxopiperidin-3-yl)-5-(((R)-pyrrolidin-3-ylmethoxy)isoindoline-1,3-dione (5-4) (140 mg, 0.39 mmol, 1.2 eq.) and TEA (200 μL, 0.96 mmol, 3.0 eq.) in DCM (2 mL) solution was stirred at room temperature for 2 hours, and then the reaction mixture was added with NaBH(OAc) 3 (85 mg, 0.39 mmol, 1.2 eq.), stirred for 16 hours, diluted with H 2 O (20 mL), and extracted with DCM (10 mL*2). The combined organic layers were washed with saturated brine, dried over anhydrous sodium sulfate, concentrated by vacuum filtration, and purified by filtration with MeCN:H 2 O=60% chromatography purification gave tert-butyl 6-(cis-4-(((3R)-3-(((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)-2-methylpiperidin-1-yl)pyridazine-3-carboxylate (26-5) (100 mg). MS: m/z=646[M+H] +

第六步：化合物26-6的制备Step 6: Preparation of compound 26-6

室温下,向6-(顺-4-(((3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)哒嗪-3-羧酸叔丁酯(26-5)(100毫克，0.15毫摩尔，1.0eq.)的DCM(2mL)溶液中加入TFA(1毫升)。搅拌6小时后，浓缩，得到6-(顺-4-(((3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)哒嗪-3-羧酸(26-6)(100毫克)。MS:m/z＝590[M+H]+ To a solution of tert-butyl 6-(cis-4-(((3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)-2-methylpiperidin-1-yl)pyridazine-3-carboxylate (26-5) (100 mg, 0.15 mmol, 1.0 eq.) in DCM (2 mL) at room temperature was added TFA (1 mL). After stirring for 6 hours, the mixture was concentrated to give 6-(cis-4-(((3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)-2-methylpiperidin-1-yl)pyridazine-3-carboxylic acid (26-6) (100 mg). MS: m/z＝590[M+H] +

第七步：化合物26的制备Step 7: Preparation of compound 26

室温下,向6-(顺-4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-氧基)甲基)吡咯烷-1-基)甲基)吡咯烷-1-基)甲基)-2-甲基哒啶-3-羧酸(26-6)(50毫克，0.08毫摩尔，1.0eq.)，5-(1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(1-7)(25毫克，0.09毫摩尔，1.1等量)，HATU(39毫克，0.10毫摩尔，1.2eq.)的DMF(2毫升)溶液加入DIEA(50微升，0.25毫摩尔，3.0eq.)。搅拌16小时后，HPLC纯化(MeCN：H2O＝55％)，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(顺-4-(3R)-3-(((2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)哒嗪-3-甲酰胺(26)(10mg)。MS:m/z＝840[M+H]+ To a solution of 6-(cis-4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-oxy)methyl)pyrrolidin-1-yl)methyl)pyrrolidin-1-yl)methyl)-2-methylpyrididine-3-carboxylic acid (26-6) (50 mg, 0.08 mmol, 1.0 eq.), 5-(1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (1-7) (25 mg, 0.09 mmol, 1.1 eq.), HATU (39 mg, 0.10 mmol, 1.2 eq.) in DMF (2 ml) was added DIEA (50 μl, 0.25 mmol, 3.0 eq.) at room temperature. After stirring for 16 hours, the mixture was purified by HPLC (MeCN: H2O =55%) to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(cis-4-(3R)-3-(((2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)-2-methylpiperidin-1-yl)pyridazine-3-carboxamide (26) (10 mg). MS: m/z=840 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),9.07(dd,J＝4.3,1.7Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8.60(d,J＝8.0Hz,1H),8.27(d,J＝8.3Hz,1H),7.91–7.80(m,2H),7.68(dd,J＝8.5,4.3Hz,1H),7.48(d,J＝6.5Hz,1H),7.42–7.29(m,3H),5.13(dd,J＝12.9,5.3Hz,1H),4.96–4.86(m,1H),4.76–4.70(m,1H),4.45–4.35(m,1H),4.33–4.16(m,3H),4.00–3.87(m,2H),3.24–3.03(m,4H),2.94–2.83(m,2H),2.66–2.53(m,2H),2.36–2.17(m,4H),2.10–2.03(m,1H),2.02–1.89(m,4H),1.83–1.63(m,5H),1.48–1.41(m,1H),1.30–1.14(m,5H). 1 H NMR (400MHz, DMSO-d 6 )δ11.11(s,1H),9.07(dd,J＝4.3,1.7Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8.60(d,J＝8.0Hz,1H), 8.27(d,J＝8.3Hz,1H),7.91–7 .80(m,2H),7.68(dd,J＝8.5,4.3Hz,1H),7.48(d,J＝6.5Hz,1H),7.42–7.29(m,3H),5.13(dd,J＝12.9 ,5.3Hz,1H),4.96–4.86(m, 1H),4.76–4.70(m,1H),4.45–4.35(m,1H),4.33–4.16(m,3H),4.00–3.87(m,2H),3.24–3.03(m,4H),2.94– 2.83(m,2H),2.66–2 .53(m,2H),2.36–2.17(m,4H),2.10–2.03(m,1H),2.02–1.89(m,4H),1.83–1.63(m,5H),1.48–1.41(m, 1H),1.30–1.14(m,5H).

实施例27化合物27的制备
Example 27 Preparation of Compound 27

第一步：化合物27-1的制备Step 1: Preparation of compound 27-1

室温下,向5-溴-2-氯-3-(三氟甲基)吡啶(1.00g，3.84mmol，1.0eq.)的DMSO(6mL)溶液中加入苄基哌嗪-1-羧酸酯(0.85g，3.84mmol，1.0eq)和K2CO3(1.59g，11.5mmol，3.0eq.)。在100℃下搅拌4小时后，冷至室温，加入水(60毫升)和EA(60毫升)。有机层浓缩。残余物用柱(PE/EA 4：1)纯化，得4-(5-溴-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-1)(1.02g)。MS:m/z＝444，446[M+H]+ To a solution of 5-bromo-2-chloro-3-(trifluoromethyl)pyridine (1.00 g, 3.84 mmol, 1.0 eq.) in DMSO (6 mL) at room temperature, benzylpiperazine-1-carboxylate (0.85 g, 3.84 mmol, 1.0 eq.) and K 2 CO 3 (1.59 g, 11.5 mmol, 3.0 eq.) were added. After stirring at 100° C. for 4 hours, the mixture was cooled to room temperature, and water (60 mL) and EA (60 mL) were added. The organic layer was concentrated. The residue was purified by column (PE/EA 4:1) to give benzyl 4-(5-bromo-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (27-1) (1.02 g). MS: m/z=444, 446 [M+H] +

第二步：化合物27-2的制备Step 2: Preparation of compound 27-2

室温下,将4-(5-溴-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-1)(900毫克，2.03毫摩尔，1.0eq.)，CuI(77毫克，0.405毫摩尔，0.2eq.)，L-脯氨酸(93毫克，0.810毫摩尔，0.4eq.)，K2CO3(420毫克，3.04毫摩尔，1.5eq.)和NH4OH(380毫克，28％，3.04毫摩尔，1.5eq.)混合在DMSO(5mL)中。在90℃下搅拌12小时后，冷至室温，加入EA(50毫升)。过滤混合物。滤液用水(50mL)洗涤浓缩，得4-(5-氨基-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-2)(810mg)。MS:m/z＝381[M+H]+ 4-(5-bromo-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid benzyl ester (27-1) (900 mg, 2.03 mmol, 1.0 eq.), CuI (77 mg, 0.405 mmol, 0.2 eq.), L-proline (93 mg, 0.810 mmol, 0.4 eq.), K 2 CO 3 (420 mg, 3.04 mmol, 1.5 eq.) and NH 4 OH (380 mg, 28%, 3.04 mmol, 1.5 eq.) were mixed in DMSO (5 mL) at room temperature. After stirring at 90°C for 12 hours, the mixture was cooled to room temperature and EA (50 mL) was added. The mixture was filtered. The filtrate was washed with water (50 mL) and concentrated to give 4-(5-amino-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid benzyl ester (27-2) (810 mg). MS: m/z = 381 [M + H] +

第三步：化合物27-3的制备Step 3: Preparation of compound 27-3

将乳酸(221mg，85％，2.08mmol，1.3eq)和DBU(317mg，2.08mmol，1.3eq)的混合物在室温下搅拌1h。加入4-(5-氨基-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-2)(610mg，1.52mmol，1.0eq)和丙烯酸甲酯(3mL)。在85℃搅拌16小时后，冷至室温，加入EA(40毫升)和水(40毫升)。有机层浓缩。残余物用柱(PE/EA 2∶1至1∶1)纯化，得到4-(5-(((3-甲氧基-3-氧代丙基)氨基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-3)(445mg)。MS:m/z＝467[M+H]+ A mixture of lactic acid (221 mg, 85%, 2.08 mmol, 1.3 eq) and DBU (317 mg, 2.08 mmol, 1.3 eq) was stirred at room temperature for 1 h. 4-(5-amino-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid benzyl ester (27-2) (610 mg, 1.52 mmol, 1.0 eq) and methyl acrylate (3 mL) were added. After stirring at 85°C for 16 hours, the mixture was cooled to room temperature and EA (40 mL) and water (40 mL) were added. The organic layer was concentrated. The residue was purified by column (PE/EA 2:1 to 1:1) to give 4-(5-(((3-methoxy-3-oxopropyl)amino)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid benzyl ester (27-3) (445 mg). MS: m/z=467[M+H] +

第四步：化合物27-4的制备Step 4: Preparation of compound 27-4

室温下,向4-(5-(3-甲氧基-3-氧代丙基)氨基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-3)(445mg，0.95mmol，1.0eq.)的AcOH(4.5mL)溶液中加入KCNO(155mg，1.91mmol，2.0eq.)。 搅拌16小时后，加入EA(50毫升)和水(50毫升)。有机层浓缩。残余物用柱(EA/MeOH 10：1)纯化，得到4-(5-(1-(3-甲氧基-3-氧代丙基)脲基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-4)(300mg)。MS:m/z＝510[M+H]+ To a solution of benzyl 4-(5-(3-methoxy-3-oxopropyl)amino)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (27-3) (445 mg, 0.95 mmol, 1.0 eq.) in AcOH (4.5 mL) was added KCNO (155 mg, 1.91 mmol, 2.0 eq.) at room temperature. After stirring for 16 hours, EA (50 mL) and water (50 mL) were added. The organic layer was concentrated. The residue was purified by column (EA/MeOH 10:1) to give benzyl 4-(5-(1-(3-methoxy-3-oxopropyl)ureido)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (27-4) (300 mg). MS: m/z=510 [M+H] +

第五步：化合物27-5的制备Step 5: Preparation of compound 27-5

室温下,向4-(5-(1-(3-(3-甲氧基-3-氧代丙基)脲基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-4)(300mg，0.59mmol，1.0eq.)的MeCN(3mL)溶液中加入Triton-B(246mg，0.59mmol，1.0eq.)。搅拌16小时后，加入EA(30mL)和水(30mL)。有机层浓缩。残余物用制备TLC(纯EA)纯化，得到4-(5-(2，4-二氧代四氢嘧啶-1(2H)-基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-5)(84mg)。MS:m/z＝478[M+H]+ Triton-B (246 mg, 0.59 mmol, 1.0 eq.) was added to a solution of benzyl 4-(5-(1-(3-(3-methoxy-3-oxopropyl)ureido)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (27-4) (300 mg, 0.59 mmol, 1.0 eq.) in MeCN (3 mL) at room temperature. After stirring for 16 hours, EA (30 mL) and water (30 mL) were added. The organic layer was concentrated. The residue was purified by preparative TLC (neat EA) to give benzyl 4-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylate (27-5) (84 mg). MS: m/z=478 [M+H] +

第六步：化合物27-6的制备Step 6: Preparation of compound 27-6

室温下,向4-(5-(2，4-二氧代四氢嘧啶-1(2H)-基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-羧酸苄酯(27-5)(84毫克，0.176毫摩尔，1.0eq.)的EA(1mL)溶液中加入Pd/C(10毫克，5％)。在H2(1个大气压)下搅拌16小时，过滤，滤液浓缩得到1-(6-(哌嗪-1-基)-5-(三氟甲基)吡啶-3-基)二氢嘧啶-2，4(1H，3H)-二酮(27-6)(67毫克)。MS:m/z＝344[M+H]+ Pd/C (10 mg, 5%) was added to a solution of 4-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-carboxylic acid benzyl ester (27-5) (84 mg, 0.176 mmol, 1.0 eq.) in EA (1 mL) at room temperature. The mixture was stirred under H 2 (1 atm) for 16 hours, filtered, and the filtrate was concentrated to give 1-(6-(piperazin-1-yl)-5-(trifluoromethyl)pyridin-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (27-6) (67 mg). MS: m/z=344[M+H] +

第七步：化合物27-7的制备Step 7: Preparation of compound 27-7

室温下,向1-(6-((哌嗪-1-基)-5-(三氟甲基)吡啶-3-基)二氢嘧啶-2，4(1H，3H)-二酮(27-6)(67mg，0.176mmol，1.0eq.)和6-(4-甲酰基哌啶-1-基)哒嗪-3-羧酸叔丁酯(51mg，0.176mmol，1.0eq.)的DCM(2mL)和DMF(1mL)溶液中加入TEA(178mg，1.76mmol，10eq.)和MgSO4(212mg，1.76毫摩尔)。搅拌2h后，加入NaBH(OAc)3(93毫克，0.44毫摩尔，2.5eq.)。搅拌1小时后，添加了DCM(20mL)。过滤混合物并浓缩滤液。加入EA(40毫升)和水(40毫升)。有机层浓缩。残余物通过制备TLC(纯MeCN)纯化，得到白色固体的6-((4-(5-(2，4-二氧代四氢嘧啶-1(2H)-基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-基)甲基)哒嗪-3-羧酸叔丁酯(27-7)(30mg)。MS:m/z＝344[M+H]+ To a solution of 1-(6-((piperazin-1-yl)-5-(trifluoromethyl)pyridin-3-yl)dihydropyrimidine-2,4(1H,3H)-dione (27-6) (67 mg, 0.176 mmol, 1.0 eq.) and tert-butyl 6-(4-formylpiperidin-1-yl)pyridazine-3-carboxylate (51 mg, 0.176 mmol, 1.0 eq.) in DCM (2 mL) and DMF (1 mL) at room temperature were added TEA (178 mg, 1.76 mmol, 10 eq.) and MgSO 4 (212 mg, 1.76 mmol). After stirring for 2 h, NaBH(OAc) 3 was added. (93 mg, 0.44 mmol, 2.5 eq.). After stirring for 1 hour, DCM (20 mL) was added. The mixture was filtered and the filtrate was concentrated. EA (40 mL) and water (40 mL) were added. The organic layer was concentrated. The residue was purified by preparative TLC (pure MeCN) to give 6-((4-(5-(2,4-dioxotetrahydropyrimidine-1(2H)-yl)-3-(trifluoromethyl)pyridin-2-yl)piperazine-1-yl)methyl)pyridazine-3-carboxylic acid tert-butyl ester (27-7) (30 mg) as a white solid. MS: m/z=344[M+H] +

第八步：化合物27-8的制备Step 8: Preparation of Compound 27-8

室温下,向6-((4-(5-(2，4-二氧代四氢嘧啶-1(2H)-基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-基)甲基)哒嗪-3-羧酸叔丁酯(27-7)(212毫克，1.76毫摩尔，10eq.)的DCM(1毫升)溶液中加入TFA(1毫升)。搅拌1小时后，浓缩得到6-(4-(5-(2，4-二氧代四氢嘧啶-1(2H)-基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-基)甲基)哒啶-1-基)哒嗪-3-羧酸(27-8)(47毫克)。MS:m/z＝563[M+H]+ At room temperature, TFA (1 ml) was added to a solution of tert-butyl 6-((4-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)methyl)pyridazine-3-carboxylate (27-7) (212 mg, 1.76 mmol, 10 eq.) in DCM (1 ml). After stirring for 1 hour, the mixture was concentrated to give 6-(4-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)methyl)pyridin-1-yl)pyridazine-3-carboxylic acid (27-8) (47 mg). MS: m/z=563[M+H] +

第九步：化合物27的制备Step 9: Preparation of compound 27

室温下,向6-(4-((4-(5-(2，4-二氧代四氢嘧啶-1(2H)-基)-3-(三氟甲基)吡啶-2-基)哌嗪-1-基)甲基)哌啶-1-基)哒嗪-3-羧酸(27-8)(47mg，0.048mmol，1.0eq.)的DMF(1mL)溶液中加入DIEA(25mg，0.192mmol，4.0eq.)和5-(((1r，4r)-4-氨基环己基)氧基)喹啉-8-甲腈(15mg，0.048mmol，1.0eq.)。慢慢加入HATU(20毫克，0.053毫摩尔，1.1eq.)。搅拌16小时后，EA(20毫升)和水(20毫升)。有机层浓缩。残余物经反相色谱法纯化，用水(0.05％TFA)和MeCN洗脱，得到N-((1r，4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-6-(4-(5-(2，4-二氧代四氢嘧啶-1(2H)-基) -3-(三氟甲基)吡啶-2-基)哌嗪-1-基)甲基)哒啶-1-基)哒嗪-3-甲酰胺(27)(14mg)。MS:m/z＝812[M+H]+ To a solution of 6-(4-((4-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl)-3-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)methyl)piperidin-1-yl)pyridazine-3-carboxylic acid (27-8) (47 mg, 0.048 mmol, 1.0 eq.) in DMF (1 mL) at room temperature were added DIEA (25 mg, 0.192 mmol, 4.0 eq.) and 5-(((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile (15 mg, 0.048 mmol, 1.0 eq.). HATU (20 mg, 0.053 mmol, 1.1 eq.) was slowly added. After stirring for 16 hours, EA (20 mL) and water (20 mL) were added. The organic layer was concentrated. The residue was purified by reverse phase chromatography eluting with water (0.05% TFA) and MeCN to afford N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-6-(4-(5-(2,4-dioxotetrahydropyrimidin-1(2H)-yl) -3-(trifluoromethyl)pyridin-2-yl)piperazin-1-yl)methyl)pyridin-1-yl)pyridazine-3-carboxamide (27) (14 mg). MS: m/z=812 [M+H] +

1H NMR(400MHz,DMSO-d6)δ10.59(s,1H),9.65(s,1H),9.08(dd,J＝4.2,1.7Hz,1H),8.67–8.60(m,2H),8.29(d,J＝8.4Hz,1H),8.17(d,J＝2.5Hz,1H),7.85(d,J＝9.5Hz,1H),7.70(dd,J＝8.5,4.2Hz,1H),7.41(d,J＝9.7Hz,1H),7.35(d,J＝8.5Hz,1H),4.76-4.72(m,1H),4.53(d,J＝13.3Hz,2H),3.98-3.93(m,1H),3.86(t,J＝6.6Hz,2H),3.68-3.63(m,2H),3.55-3.49(m,2H),3.36(t,J＝12.4Hz,2H),3.19–3.04(m,6H),2.75(t,J＝6.6Hz,2H),2.26-2.24(m,3H),1.99-1.97(m,2H),1.91-1.86(m,2H),1.73-1.68(m,3H),1.31-1.22(m,3H). 1 H NMR (400MHz, DMSO-d6) δ10.59(s,1H),9.65(s,1H),9.08(dd,J＝4.2,1.7Hz,1H),8.67–8.60(m,2H),8.29(d ,J＝8.4Hz,1H),8.17(d,J ＝2.5Hz,1H),7.85(d,J＝9.5Hz,1H),7.70(dd,J＝8.5,4.2Hz,1H),7.41(d,J＝9.7Hz,1H),7.35(d,J ＝8.5Hz,1H),4.76-4.72(m,1H),4. 53(d,J＝13.3Hz,2H),3.98-3.93(m,1H),3.86(t,J＝6.6Hz,2H),3.68-3.63(m,2H),3.55-3.49(m,2H) ,3.36(t,J＝12.4Hz,2H),3.19– 3.04(m,6H),2.75(t,J＝6.6Hz,2H),2.26-2.24(m,3H),1.99-1.97(m,2H),1.91-1.86(m,2H),1.73-1.68( m,3H),1.31-1.22(m,3H).

实施例28化合物28的制备
Example 28 Preparation of Compound 28

第一步：化合物28-1的制备Step 1: Preparation of compound 28-1

在0℃下,向叔丁基((1r，3r)-3-羟基-2，2，4，4-四甲基环丁基)氨基甲酸叔丁酯(350mg，1.44mmol，1.00eq.)的DMF(5mL)溶液中加入氢化钠(115mg，2.88mmol，60％重量，2.00eq.)，然后滴加4-氟-2-甲氧基苯甲苯(239mg，1.58mmol，1.10eq.)的DMF(10mL)溶液。在室温并搅拌3.5小时，加入饱和氯化铵水溶液(30mL)淬灭，并用乙酸乙酯(30mL)萃取。用饱和食盐水(30mL×2)洗涤有机层，用硫酸钠干燥并减压浓缩，得到残余物。残余物在硅胶(石油醚：乙酸乙酯＝20：1)上柱层析纯化，得到叔丁基((1r，3r)-3-(4-氰基-3-甲氧基苯氧基)-2，2，4，4-四甲基环丁基)氨基甲酸酯(28-1)(258mg)。At 0°C, sodium hydride (115 mg, 2.88 mmol, 60% weight, 2.00 eq.) was added to a solution of tert-butyl ((1r, 3r)-3-hydroxy-2,2,4,4-tetramethylcyclobutyl)carbamate (350 mg, 1.44 mmol, 1.00 eq.) in DMF (5 mL), and then a solution of 4-fluoro-2-methoxybenzene toluene (239 mg, 1.58 mmol, 1.10 eq.) in DMF (10 mL) was added dropwise. The mixture was stirred at room temperature for 3.5 hours, quenched with saturated aqueous ammonium chloride solution (30 mL), and extracted with ethyl acetate (30 mL). The organic layer was washed with saturated brine (30 mL × 2), dried over sodium sulfate and concentrated under reduced pressure to obtain a residue. The residue was purified by column chromatography on silica gel (petroleum ether:ethyl acetate=20:1) to give tert-butyl ((1r,3r)-3-(4-cyano-3-methoxyphenoxy)-2,2,4,4-tetramethylcyclobutyl)carbamate (28-1) (258 mg).

MS:m/z＝374[M+H]+ MS: m/z = 374 [M + H] +

第二步：化合物28-2的制备Step 2: Preparation of compound 28-2

室温下,向叔丁基((1r，3r)-3-(4-氰基-3-甲氧基苯氧基)-2，2，4，4-四甲基环丁基)氨基甲酸叔丁酯(258mg，1.44mmol，1.00eq.)的DCM(5mL)中，滴入4M HCl in dioxane(3mL)。搅拌6小时后，浓缩，得到4-((1r，3r)-3-氨基-2，2，4，4-四甲基环丁氧基)-2-甲氧基苯腈(28-2)(240毫克)。MS:m/z＝274[M+H]+ At room temperature, 4M HCl in dioxane (3 mL) was added dropwise to tert-butyl ((1r, 3r)-3-(4-cyano-3-methoxyphenoxy)-2,2,4,4-tetramethylcyclobutyl)carbamate (258 mg, 1.44 mmol, 1.00 eq.) in DCM (5 mL). After stirring for 6 hours, the mixture was concentrated to give 4-((1r, 3r)-3-amino-2,2,4,4-tetramethylcyclobutyloxy)-2-methoxybenzonitrile (28-2) (240 mg). MS: m/z=274 [M+H] +

第一步：化合物28-3的制备 Step 1: Preparation of compound 28-3

室温下,向2-(2，6-二氧代哌啶-3-基)-5-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-1，3-二酮(5-4)(100毫克，0.28毫摩尔，1.0eq.)，4-(4-甲酰基哌啶-1-基)苯甲酸叔丁酯(81毫克，0.28毫摩尔，1.0eq.)的DCM(2毫升)中加入TEA(0.2毫升)。搅拌1小时后，加入NaBH(OAc)3(71mg，0.33mmol，1.2eq.)。继续搅拌16h后，浓缩。HPLC纯化(MeCN：H2O＝70％)得到4-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(28-3)(80mg)。MS:m/z＝630[M+H]+ To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-(((R)-pyrrolidin-3-yl)methoxy)isoindoline-1,3-dione (5-4) (100 mg, 0.28 mmol, 1.0 eq.), tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (81 mg, 0.28 mmol, 1.0 eq.) in DCM (2 ml) was added TEA (0.2 ml) at room temperature. After stirring for 1 hour, NaBH(OAc) 3 (71 mg, 0.33 mmol, 1.2 eq.) was added. Stirring was continued for 16 hours and then concentrated. HPLC purification (MeCN: H2O =70%) gave tert-butyl 4-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoate (28-3) (80 mg). MS: m/z=630 [M+H] +

第二步：化合物28-4的制备Step 2: Preparation of compound 28-4

室温下,向4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯(28-3)(80毫克，0.12毫摩尔，1.0eq.)的DCM(2毫升)中加入TFA(1毫升)。搅拌16小时后，浓缩，得到4-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸(28-4)(80毫克)。MS:m/z＝574[M+H]+ To tert-butyl 4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoate (28-3) (80 mg, 0.12 mmol, 1.0 eq.) in DCM (2 ml) was added TFA (1 ml) at room temperature. After stirring for 16 hours, the mixture was concentrated to give 4-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoic acid (28-4) (80 mg). MS: m/z=574[M+H] +

第三步：化合物28的制备Step 3: Preparation of compound 28

室温下,向4-(4-(3R)-3-(((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸(28-4)(80毫克，0.14毫摩尔，1.0eq.)，4-((1r，3r)-3-氨基-2，2，4，4-四甲基环丁氧基)-2-甲氧基苯腈(38毫克，0.14毫摩尔，1.0eq.)，HATU(63毫克，0.16毫摩尔，1.2eq.)的DMF(2毫升)中的溶液加入DIEA(0.1毫升，0.42毫摩尔，3.0eq.)。搅拌16小时后，浓缩，HPLC纯化(MeCN：H2O＝55％)，得到N-((1r，3r)-3-(4-氰基-3-甲氧基苯氧基)-2，2，4，4-四甲基环丁基)-4-(4-(3R)-3-((2-(2，6-二氧代哌啶-3-基)-1，3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(28)(22mg)。MS:m/z＝831[M+H]+ To a solution of 4-(4-(3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoic acid (28-4) (80 mg, 0.14 mmol, 1.0 eq.), 4-((1r,3r)-3-amino-2,2,4,4-tetramethylcyclobutyloxy)-2-methoxybenzonitrile (38 mg, 0.14 mmol, 1.0 eq.), HATU (63 mg, 0.16 mmol, 1.2 eq.) in DMF (2 ml) was added DIEA (0.1 ml, 0.42 mmol, 3.0 eq.) at room temperature. After stirring for 16 hours, the mixture was concentrated and purified by HPLC (MeCN: H2O =55%) to give N-((1r,3r)-3-(4-cyano-3-methoxyphenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-(3R)-3-((2-(2,6-dioxopiperidin-3-yl)-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (28) (22 mg). MS: m/z=831 [M+H] +

1H NMR(400MHz,DMSO-d6)δ11.08(s,1H),7.81(d,J＝8.3Hz,1H),7.70(d,J＝8.5Hz,2H),7.62(d,J＝8.6Hz,1H),7.49–7.41(m,2H),7.34(dd,J＝8.3,2.2Hz,1H),6.92(d,J＝8.6Hz,2H),6.61(d,J＝2.2Hz,1H),6.51(dd,J＝8.7,2.2Hz,1H),5.09(dd,J＝13.0,5.4Hz,1H),4.24(s,1H),4.09–3.99(m,3H),3.88(s,3H),3.85–3.77(m,2H),3.48(s,1H),2.93–2.81(m,2H),2.78–2.68(m,2H),2.63–2.52(m,4H),2.41–2.22(m,4H),2.04–1.89(m,3H),1.83–1.74(m,2H),1.20(s,6H),1.12(s,6H). 1 H NMR (400MHz, DMSO-d 6 )δ11.08(s,1H),7.81(d,J＝8.3Hz,1H),7.70(d,J＝8.5Hz,2H),7.62(d,J＝8.6Hz,1H),7.49–7.41( m,2H),7.34(dd, J＝8.3,2.2Hz,1H),6.92(d,J＝8.6Hz,2H),6.61(d,J＝2.2Hz,1H),6.51(dd,J＝8.7,2.2Hz,1H),5.09( dd,J＝13.0,5. 4Hz,1H),4.24(s,1H),4.09–3.99(m,3H),3.88(s,3H),3.85–3.77(m,2H),3.48(s,1H),2.93–2.81(m, 2H),2.78–2 .68(m,2H),2.63–2.52(m,4H),2.41–2.22(m,4H),2.04–1.89(m,3H),1.83–1.74(m,2H),1.20(s,6H) ,1.12(s,6H).

实施例29化合物29的制备
Example 29 Preparation of Compound 29

室温下,向4-羟基苯甲酸甲酯(3.33g，21.8mmol，1.10eq.)和叔丁基(R)-3-(羟甲基)吡咯烷-1-羧酸叔丁酯(4.00g，19.8mmol，1.00eq.)的THF(30mL)溶液中，加入PPh3(6.26g，23.8mmol，1.20eq.)，然后将混合物冷却至0℃，并加入DIAD(5.02g，24.8mmol，1.25eq.)，将混合物在25℃下搅拌16小时。将混合物减压浓缩，粗产物用硅胶柱层析(PE：EtOAc＝10：1)纯化，得到白色固体的叔丁基(R)-3-((4-(甲氧基羰基)苯氧基)甲基)吡咯烷-1-羧酸酯(29-3)(7.00克)。MS:m/z＝335[M+H]+ To a solution of methyl 4-hydroxybenzoate (3.33 g, 21.8 mmol, 1.10 eq.) and tert-butyl (R)-3-(hydroxymethyl)pyrrolidine-1-carboxylate (4.00 g, 19.8 mmol, 1.00 eq.) in THF (30 mL) at room temperature, PPh 3 (6.26 g, 23.8 mmol, 1.20 eq.) was added, and the mixture was cooled to 0° C., DIAD (5.02 g, 24.8 mmol, 1.25 eq.) was added, and the mixture was stirred at 25° C. for 16 hours. The mixture was concentrated under reduced pressure, and the crude product was purified by silica gel column chromatography (PE: EtOAc = 10: 1) to give tert-butyl (R)-3-((4-(methoxycarbonyl)phenoxy)methyl)pyrrolidine-1-carboxylate (29-3) (7.00 g) as a white solid. MS: m/z = 335 [M + H] +

将氢氧化钠(2.98g，74.5mmol，5.00eq.)加入到叔丁基(R)-3-((4-(甲氧基羰基)苯氧基)甲基)吡咯烷-1-羧酸叔丁酯(5.00g，14.1mmol，1.00eq.)的MeOH(10mL)和H2O(10mL)溶液中。室温搅拌6小时，水相用EtOAc(50mL×3)洗涤，然后用1N HCl酸化至pH＝3，所得溶液用EtOAc(50mL×3)萃取，分离出的有机层用Na2SO4干燥，蒸发至干，得(R)-4-((1-(叔丁氧羰基)吡咯烷-3-基)甲氧基)苯甲酸(29-4)(1.00g)。MS:m/z＝321[M+H]+ Sodium hydroxide (2.98 g, 74.5 mmol, 5.00 eq.) was added to a solution of tert-butyl (R)-3-((4-(methoxycarbonyl)phenoxy)methyl)pyrrolidine-1-carboxylate (5.00 g, 14.1 mmol, 1.00 eq.) in MeOH (10 mL) and H 2 O (10 mL). The mixture was stirred at room temperature for 6 hours, the aqueous phase was washed with EtOAc (50 mL×3), and then acidified to pH=3 with 1N HCl. The resulting solution was extracted with EtOAc (50 mL×3), the separated organic layer was dried over Na 2 SO 4 , and evaporated to dryness to give (R)-4-((1-(tert-butyloxycarbonyl)pyrrolidin-3-yl)methoxy)benzoic acid (29-4) (1.00 g). MS: m/z=321[M+H] +

室温下，向(R)-4-((1-(叔丁氧羰基)吡咯烷-3-基)甲氧基)苯甲酸(1.00g，3.11mmol，1.00eq.)的乙腈(20mL)溶液中加入NMI(1.28g，15.1mmol，5.00eq.)，TCFH(1.75g，6.22mmol，2.00eq.)和3-氨基哌啶-2，6-二酮(438mg，3.42mmol，1.10eq.)。搅拌16小时后。加入乙酸乙酯(50mL)和NaHCO3水溶液(50mL)，分离的有机层用水洗涤，用Na2SO4干燥并蒸发至干，得到叔丁基(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-羧酸酯(29-5)(400mg)。MS:m/z＝431[M+H]+ To a solution of (R)-4-((1-(tert-butoxycarbonyl)pyrrolidin-3-yl)methoxy)benzoic acid (1.00 g, 3.11 mmol, 1.00 eq.) in acetonitrile (20 mL) were added NMI (1.28 g, 15.1 mmol, 5.00 eq.), TCFH (1.75 g, 6.22 mmol, 2.00 eq.) and 3-aminopiperidine-2,6-dione (438 mg, 3.42 mmol, 1.10 eq.) at room temperature, and the mixture was stirred for 16 hours. Ethyl acetate (50 mL) and aqueous NaHCO 3 solution (50 mL) were added, and the separated organic layer was washed with water, dried over Na 2 SO 4 and evaporated to dryness to give tert-butyl (3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidine-1-carboxylate (29-5) (400 mg). MS: m/z=431 [M+H] +

室温下，向叔丁基(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-羧酸叔丁酯29-5(260mg，402μmol，1.00eq.)的DCM(3mL)溶液中，滴入4M HCl in dioxane(3mL)。搅拌6小时后，浓缩，得到N-(2，6-二氧代哌啶-3-基)-4-((R)-吡咯烷-3-基)甲氧基)苯甲酰胺(29-6)(200毫克)。MS:m/z＝331[M+H]+ To a solution of tert-butyl (3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidine-1-carboxylate 29-5 (260 mg, 402 μmol, 1.00 eq.) in DCM (3 mL) at room temperature, 4M HCl in dioxane (3 mL) was added dropwise. After stirring for 6 hours, the mixture was concentrated to give N-(2,6-dioxopiperidin-3-yl)-4-((R)-pyrrolidin-3-yl)methoxy)benzamide (29-6) (200 mg). MS: m/z=331[M+H] +

室温下，向N-(2，6-二氧代哌啶-3-基)-4-(((R)-吡咯烷-3-基)甲氧基)苯甲酰胺29-6(200mg，543μmol，1.00eq.)和4-(4-甲酰基哌啶-1-基)苯甲酸叔丁酯(157mg，543μmol，1.00eq.)的DCM(10mL)溶液中加入TEA(275mg，2.72mmol，5.00eq.)。搅拌2小时后，加入NaBH(OAc)3(345mg，1.63毫摩尔，3.00eq.)。继续搅拌16小时后，浓缩，残余物通过反相色谱(TFA缓冲液)纯化，得到4-(4-(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸酯(29-7)(278mg)。MS:m/z＝604[M+H]+ To a solution of N-(2,6-dioxopiperidin-3-yl)-4-(((R)-pyrrolidin-3-yl)methoxy)benzamide 29-6 (200 mg, 543 μmol, 1.00 eq.) and tert-butyl 4-(4-formylpiperidin-1-yl)benzoate (157 mg, 543 μmol, 1.00 eq.) in DCM (10 mL) was added TEA (275 mg, 2.72 mmol, 5.00 eq.) at room temperature. After stirring for 2 hours, NaBH(OAc) 3 (345 mg, 1.63 mmol, 3.00 eq.) was added. After stirring for 16 hours, the mixture was concentrated and the residue was purified by reverse phase chromatography (TFA buffer) to give 4-(4-(3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoate (29-7) (278 mg). MS: m/z=604 [M+H] +

室温下，向4-(4-(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸叔丁酯29-7(270mg，446μmol，1.00eq.)的DCM(4mL)溶液中加入TFA(2mL)。搅拌6小时，浓缩，得到4-(4-(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸(29-8)(232毫克)。MS:m/z＝548[M+H]+ To a solution of tert-butyl 4-(4-(3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoate 29-7 (270 mg, 446 μmol, 1.00 eq.) in DCM (4 mL) at room temperature was added TFA (2 mL). The mixture was stirred for 6 hours and concentrated to give 4-(4-(3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoic acid (29-8) (232 mg). MS: m/z=548[M+H] +

室温下，向4-(4-(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酸29-8(100毫克，182微摩尔，1.00eq.)的乙腈溶液中，加入TCFH(102毫克，364微摩尔，2.00eq.)和NMI(74.8毫克，911微摩尔，5.00eq.)。搅拌1小时后，加入4-((1r，3r)-3-氨基-2，2，4，4-四甲基环丁氧基)-2-甲氧基苯腈(28-2,57.2毫克，184微摩尔，1.01eq.)。继续搅拌16小时后，浓缩至干，得到粗品，残余物经反相色谱(TFA缓冲液)纯化，得到N-((1r，3r)-3-(4-氰基-3-甲氧基苯氧基)-2，2，4，4-四甲基环丁基)-4-(4-(3R)-3-((4-((2，6-二氧代哌啶-3-基)氨基甲酰基)苯氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(29)(43.0毫克)MS:m/z＝805[M+H]+ To a solution of 4-(4-(3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzoic acid 29-8 (100 mg, 182 μmol, 1.00 eq.) in acetonitrile at room temperature, TCFH (102 mg, 364 μmol, 2.00 eq.) and NMI (74.8 mg, 911 μmol, 5.00 eq.) were added. After stirring for 1 hour, 4-((1r,3r)-3-amino-2,2,4,4-tetramethylcyclobutyloxy)-2-methoxybenzonitrile (28-2, 57.2 mg, 184 μmol, 1.01 eq.) was added. After stirring for 16 hours, the mixture was concentrated to dryness to give a crude product. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r,3r)-3-(4-cyano-3-methoxyphenoxy)-2,2,4,4-tetramethylcyclobutyl)-4-(4-(3R)-3-((4-((2,6-dioxopiperidin-3-yl)carbamoyl)phenoxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (29) (43.0 mg) MS: m/z=805 [M+H] +

1H NMR(400MHz,DMSO)δ10.84(s,1H),9.61(br,1H),7.88(d,J＝8.6Hz,2H),7.76(d,J＝8.5Hz,2H),7.65(d,J＝8.8Hz,1H),7.50(d,J＝9.3Hz,1H),7.05(d,J＝8.6Hz,2H),6.98(d,J＝8.8Hz,2H),6.63(s,1H),6.54(d,J＝10.4Hz,1H),4.79–4.72(m,1H),4.27(s,1H),4.12–4.01(m,4H),3.91(s,3H),3.88–3.84(m,3H),3.77–3.65(m,2H),3.57–3.51(m,1H),3.39–3.31(m,1H),3.04–2.91(m,2H),2.85–2.75(m,4H),2.20–2.09(m,2H),2.00–1.94(m,2H),1.86–1.79(m,2H),1.36–1.27(m,2H),1.22(s,6H),1.15(s,6H). 1 H NMR (400MHz, DMSO) δ10.84(s,1H),9.61(br,1H),7.88(d,J＝8.6Hz,2H),7.76(d,J＝8.5Hz,2H),7.65(d, J＝8.8Hz,1H),7.50(d, J＝9.3Hz,1H),7.05(d,J＝8.6Hz,2H),6.98(d,J＝8.8Hz,2H),6.63(s,1H),6.54(d,J＝10.4Hz,1H) ,4.79–4.72(m,1H),4.27(s,1H ),4.12–4.01(m,4H),3.91(s,3H),3.88–3.84(m,3H),3.77–3.65(m,2H),3.57–3.51(m,1H),3.39–3.31(m ,1H),3.04–2.91(m, 2H),2.85–2.75(m,4H),2.20–2.09(m,2H),2.00–1.94(m,2H),1.86–1.79(m,2H),1.36–1.27(m,2H),1.22( s,6H),1.15(s,6H).

实施例30化合物40的制备
Example 30 Preparation of Compound 40

在0℃下，向1-甲基-1H-吲唑-5-胺(3g，20.3mmol，1.0eq.)于H2O/98％ H2SO4＝1:1(40 mL)中的悬浮液中缓慢地添加NaNO2(1.4g，20.3mmol，1.0eq.)。将反应混合物在120℃下搅拌2小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将残余物通过硅胶柱色谱(DCM/MeOH＝20:1)纯化，得到呈黄色油状的1-甲基-1H-吲唑-5-醇(40-2)(752mg，产率：25％)。MS:m/z＝149[M+H]+ At 0°C, 1-methyl-1H-indazol-5-amine (3 g, 20.3 mmol, 1.0 eq.) was added to H 2 O/98% H 2 SO 4 = 1:1 (40 To the suspension in 40-20 mL (10% by volume) was added NaNO 2 (1.4 g, 20.3 mmol, 1.0 eq.) slowly. The reaction mixture was stirred at 120 ° C for 2 hours. After the reaction was completed, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3). The combined organic layer was dried over Na 2 SO 4 and concentrated. The residue was purified by silica gel column chromatography (DCM/MeOH=20:1) to give 1-methyl-1H-indazole-5-ol (40-2) (752 mg, yield: 25%) as a yellow oil. MS: m/z=149[M+H] +

向1-甲基-1H-吲唑-5-醇(40-2)(752mg，5.07mmol，1.0eq.)和(R)-3-((甲苯磺酰基氧基)甲基)吡咯烷-1-甲酸叔丁酯(1.8g，5.07mmol，1.0eq.)于DMF(10mL)中的溶液中添加K2CO3(2.1g，15.21mmol，3.0eq.)。将反应混合物在80℃下搅拌16小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将残余物通过硅胶柱色谱(PE/EA＝4:1)纯化，得到呈黄色油状的(R)-3-(((1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-3)(1.5g，产率：89％)。MS:m/z＝332[M+H]+。To a solution of 1-methyl-1H-indazol-5-ol (40-2) (752 mg, 5.07 mmol, 1.0 eq.) and (R)-tert-butyl 3-((tosyloxy)methyl)pyrrolidine-1-carboxylate (1.8 g, 5.07 mmol, 1.0 eq.) in DMF (10 mL) was added K 2 CO 3 (2.1 g, 15.21 mmol, 3.0 eq.). The reaction mixture was stirred at 80° C. for 16 h. After the reaction was completed, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The residue was purified by silica gel column chromatography (PE/EA=4:1) to give (R)-tert-butyl 3-(((1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-3) (1.5 g, yield: 89%) as a yellow oil. MS: m/z=332 [M+H] + .

向(R)-3-(((1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-3)(1.5g，4.52mmol，1.0eq.)于DMF(10mL)中的溶液中添加NIS(2.5g，11.3mmol，2.5eq.)。将反应混合物在50℃下搅拌4小时。在反应完成后，将混合物通过反相色谱用纯乙腈洗脱纯化，得到呈棕色油状的(R)-3-(((3-碘-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-4)(1.98g，产率：96％)。MS:m/z＝402[M+H-56]+。To a solution of (R)-tert-butyl 3-(((1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-3) (1.5 g, 4.52 mmol, 1.0 eq.) in DMF (10 mL) was added NIS (2.5 g, 11.3 mmol, 2.5 eq.). The reaction mixture was stirred at 50 °C for 4 hours. After the reaction was completed, the mixture was purified by reverse phase chromatography eluting with pure acetonitrile to give (R)-tert-butyl 3-(((3-iodo-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-4) (1.98 g, yield: 96%) as a brown oil. MS: m/z=402[M+H-56] + .

向(R)-3-(((3-碘-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-4)(1.2g，2.62mmol，1.0eq.)、2,6-双(苄氧基)-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊-2-基)吡啶(1.42g，3.41mmol，1.3eq.)和Cs2CO3(2.56g，7.86mmol，3.0eq.)于二恶烷(12mL)和H2O(3mL)中的溶液中添加Pd(dppf)Cl2(213mg，0.262mmol，0.1eq.)，将反应混合物在100℃下搅拌16小时。在反应完成后，将反应物冷却，通过硅藻土床过滤，用乙酸乙酯洗涤。分离有机层，经硫酸钠干燥并浓缩。将粗物质通过硅胶色谱用PE/EA(10:1)洗脱纯化，得到呈棕色油状的(R)-3-(((3-(2,6-双(苄氧基)吡啶-3-基)-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-5)(1.4g，产率：86％)。MS:m/z＝621[M+H]+。To a solution of (R)-tert-butyl 3-(((3-iodo-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-4) (1.2 g, 2.62 mmol, 1.0 eq.), 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine (1.42 g, 3.41 mmol, 1.3 eq.) and Cs2CO3 (2.56 g, 7.86 mmol, 3.0 eq.) in dioxane (12 mL) and H2O (3 mL) was added Pd(dppf) Cl2 (213 mg, 0.262 mmol, 0.1 eq.) and the reaction mixture was stirred at 100°C for 16 hours. After completion of the reaction, the reactant was cooled, filtered through a celite bed, and washed with ethyl acetate. The organic layer was separated, dried over sodium sulfate and concentrated. The crude material was purified by silica gel chromatography eluting with PE/EA (10:1) to give (R)-tert-butyl 3-(((3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-5) (1.4 g, yield: 86%) as a brown oil. MS: m/z=621 [M+H] + .

向(R)-3-(((3-(2,6-双(苄氧基)吡啶-3-基)-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-5)(500mg，0.805mmol，1.0eq.)和10％ Pd/C(100mg，20重量％)于EtOH(5mL)中的溶液中添加(Boc)2O(175mg，0.805mmol，1.0eq.)，将反应混合物在室温下在H2(50Psi)下搅拌16小时。在反应完成后，过滤混合物并真空浓缩，得到呈棕色油状的(3R)-3-(((3-(2,6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-6)(351mg，粗物质)。MS:m/z＝343[M+H-100]+。To a solution of (R)-tert-butyl 3-(((3-(2,6-bis(benzyloxy)pyridin-3-yl)-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-5) (500 mg, 0.805 mmol, 1.0 eq.) and 10% Pd/C (100 mg, 20 wt%) in EtOH (5 mL) was added (Boc) 2O (175 mg, 0.805 mmol, 1.0 eq.) and the reaction mixture was stirred at room temperature under H2 (50 Psi) for 16 h. After the reaction was complete, the mixture was filtered and concentrated in vacuo to give tert-butyl (3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-6) (351 mg, crude) as a brown oil. MS: m/z=343 [M+H-100] + .

向(3R)-3-(((3-(2,6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(40-6)(305mg，0.8mmol，1.0eq.)于EA(2mL)中的溶液中添加4M HCl/EA(3mL)，将反应混合物在室温下搅拌6小时。在反应完成后，真空浓缩混合物，得到呈黑色固体状的3-(1-甲基-5-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2,6-二酮盐酸盐(40-7)(252mg，粗物质)。MS:m/z＝343[M+H]+。 To a solution of tert-butyl (3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (40-6) (305 mg, 0.8 mmol, 1.0 eq.) in EA (2 mL) was added 4M HCl/EA (3 mL), and the reaction mixture was stirred at room temperature for 6 hours. After the reaction was complete, the mixture was concentrated in vacuo to give 3-(1-methyl-5-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione hydrochloride (40-7) (252 mg, crude) as a black solid. MS: m/z=343[M+H] + .

向3-(1-甲基-5-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2,6-二酮盐酸盐(40-7)(80mg，0.21mmol，1.0eq.)和N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(101mg，0.21mmol，1.0eq.)于DMF(2mL)中的溶液中添加TEA(85mg，0.84mmol，4.0eq.)，将反应混合物在室温下搅拌4小时，然后向混合物中添加NaBH(OAc)3(69mg，0.31mmol，1.5eq.)并在室温下搅拌4小时。在反应完成后，将混合物通过反相色谱用MeCN:H2O＝50％洗脱纯化，得到呈白色固体状的N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(40)(29.2mg，产率：17％)。MS:m/z＝810[M+H]+ To a solution of 3-(1-methyl-5-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione hydrochloride (40-7) (80 mg, 0.21 mmol, 1.0 eq.) and N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (101 mg, 0.21 mmol, 1.0 eq.) in DMF (2 mL) was added TEA (85 mg, 0.84 mmol, 4.0 eq.), the reaction mixture was stirred at room temperature for 4 hours, and then NaBH(OAc) 3 (69 mg, 0.31 mmol, 1.5 eq.) was added to the mixture and stirred at room temperature for 4 hours. After the reaction was completed, the mixture was purified by reverse phase chromatography eluting with MeCN:H 2 O=50% to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (40) (29.2 mg, yield: 17%) as a white solid. MS: m/z=810 [M+H] +

1H NMR(400MHz,DMSO-d6)δ10.84(s,1H),9.48(brs,1H),9.05(dd,J＝4.2,1.7Hz,1H),8.62(dd,J＝8.6,1.6Hz,1H),8.27(d,J＝8.3Hz,1H),7.97(d,J＝7.2Hz,1H),7.73(d,J＝8.0Hz,2H),7.67(dd,J＝8.5,4.3Hz,1H),7.52(d,J＝9.1Hz,1H),7.33(d,J＝8.4Hz,1H),7.15(s,1H),7.12–7.02(m,1H),6.94(d,J＝7.1Hz,2H),4.71(brs,1H),4.30(dd,J＝9.6,4.9Hz,1H),4.17–4.04(m,1H),4.03–3.91(m,4H),3.86(d,J＝11.0Hz,3H),3.73–3.59(m,2H),3.19–3.09(m,2H),3.03–2.88(m,2H),2.77(t,J＝10.6Hz,2H),2.71–2.57(m,2H),2.41–2.30(m,1H),2.29–2.21(m,2H),2.21–2.12(m,1H),1.99–1.88(m,3H),1.88–1.71(m,2H),1.71–1.51(m,4H),1.26(dd,J＝20.6,11.1Hz,2H). 1 H NMR (400 MHz, DMSO-d 6 )δ10.84(s,1H),9.48(brs,1H),9.05(dd,J＝4.2,1.7Hz,1H),8.62(dd,J＝8.6,1 .6Hz,1H),8.27(d,J＝8.3Hz,1H),7.97(d,J＝7.2Hz,1H),7.73(d,J＝8.0Hz,2H),7 .67(dd,J＝8.5,4.3Hz,1H),7.52(d,J＝9.1Hz,1H),7.33(d,J＝8.4Hz,1H),7.15(s ,1H),7.12–7.02(m,1H),6.94(d,J＝7.1Hz,2H),4.71(brs,1H),4.30(dd,J＝9.6, 4.9Hz,1H),4.17–4.04(m,1H),4.03–3.91(m,4H),3.86(d,J＝11.0Hz,3H),3.73– 3.59(m,2H),3.19–3.09(m,2H),3.03–2.88(m,2H),2.77(t,J＝10.6Hz,2H),2.71 –2.57(m,2H),2.41–2.30(m,1H),2.29–2.21(m,2H),2.21–2.12(m,1H),1.99–1. 88(m,3H),1.88–1.71(m,2H),1.71–1.51(m,4H),1.26(dd,J＝20.6,11.1Hz,2H).

实施例31化合物43的制备
Example 31 Preparation of Compound 43

向2-(2,6-二氧代哌啶-3-基)-5,6-二氟异吲哚啉-1,3-二酮(43-1)(800mg，2.72mmol，1.0eq.)和Cs2CO3(2.06g，5.44mmol，2.0eq.)于乙腈(8mL)中的溶液中添加(R)-3-(羟甲基)吡咯烷-1-甲酸叔丁酯(950mg，5.0mmol，1.0eq.)。将反应混合物在100℃下搅拌16小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×2)萃取。将合并的有机层用盐水洗涤，经无水硫酸钠干燥并蒸发。将残余物通过硅胶色谱用PE/EA(1:1)洗脱纯化，得到呈白色固体状的(3R)-3-(((2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(43-2)(41mg，3.1％产率)。MS:m/z＝420[M+H-56]+。To a solution of 2-(2,6-dioxopiperidin-3-yl)-5,6-difluoroisoindoline-1,3-dione (43-1) (800 mg, 2.72 mmol, 1.0 eq.) and Cs 2 CO 3 (2.06 g, 5.44 mmol, 2.0 eq.) in acetonitrile (8 mL) was added (R)-tert-butyl 3-(hydroxymethyl)pyrrolidine-1-carboxylate (950 mg, 5.0 mmol, 1.0 eq.). The reaction mixture was stirred at 100° C. for 16 hours. After the reaction was complete, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×2). The combined organic layers were washed with brine, dried over anhydrous sodium sulfate and evaporated. The residue was purified by silica gel chromatography eluting with PE/EA (1:1) to give (3R)-tert-butyl 3-(((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (43-2) (41 mg, 3.1% yield) as a white solid. MS: m/z=420 [M+H-56] + .

向(3R)-3-(((2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(43-2)(41mg，0.086mmol，1.0eq.)于DCM(1mL)中的溶液中添加二恶烷中的4M HCl(2mL)。将混合物在室温下搅拌2小时。在反应完成后。减压浓缩反应混合物，得到呈白色固体状的2-(2,6-二氧代哌啶-3-基)-5-氟-6-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-1,3-二酮盐酸 盐(43-3)(31mg，粗物质)。MS:m/z＝376[M+H]+。To a solution of tert-butyl (3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (43-2) (41 mg, 0.086 mmol, 1.0 eq.) in DCM (1 mL) was added 4M HCl in dioxane (2 mL). The mixture was stirred at room temperature for 2 hours. After the reaction was complete. The reaction mixture was concentrated under reduced pressure to give 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-6-(((R)-pyrrolidin-3-yl)methoxy)isoindolin-1,3-dione hydrochloride as a white solid. Salt (43-3) (31 mg, crude material). MS: m/z=376 [M+H] + .

向2-(2,6-二氧代哌啶-3-基)-5-氟-6-(((R)-吡咯烷-3-基)甲氧基)异吲哚啉-1,3-二酮盐酸盐(43-3)(31mg，0.07mmol，1.0eq.)于DMF(1mL)中的溶液中添加N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(37mg，0.07mmol，1.0eq.)和TEA(30mg，0.3mmol，4.0eq.)。将反应混合物在室温下搅拌4小时，然后添加NaBH(OAc)3(16mg，0.07mmol，1.0eq.)。将反应混合物在室温下搅拌12小时。LCMS显示反应完成，MS 842。将残余物通过反相色谱用水(0.05％ TFA)和MeCN洗脱纯化，得到呈白色固体状的N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3R)-3-(((2-(2,6-二氧代哌啶-3-基)-6-氟-1,3-二氧代异吲哚啉-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(43)(10.8mg，17.1％产率)。MS:m/z＝842[M+H]+。To a solution of 2-(2,6-dioxopiperidin-3-yl)-5-fluoro-6-(((R)-pyrrolidin-3-yl)methoxy)isoindoline-1,3-dione hydrochloride (43-3) (31 mg, 0.07 mmol, 1.0 eq.) in DMF (1 mL) was added N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (37 mg, 0.07 mmol, 1.0 eq.) and TEA (30 mg, 0.3 mmol, 4.0 eq.). The reaction mixture was stirred at room temperature for 4 hours, then NaBH(OAc) 3 (16 mg, 0.07 mmol, 1.0 eq.) was added. The reaction mixture was stirred at room temperature for 12 hours. LCMS showed the reaction was complete, MS 842. The residue was purified by reverse phase chromatography eluting with water (0.05% TFA) and MeCN to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3R)-3-(((2-(2,6-dioxopiperidin-3-yl)-6-fluoro-1,3-dioxoisoindolin-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (43) (10.8 mg, 17.1% yield) as a white solid. MS: m/z = 842 [M+H] + .

1H NMR(400MHz,DMSO-d6)δ11.10(s,1H),9.52(s,1H),9.05(dd,J＝4.4,1.7Hz,1H),8.62(dd,J＝8.6,1.7Hz,1H),8.26(d,J＝8.3Hz,1H),7.98(d,J＝7.6Hz,1H),7.89(d,J＝9.2Hz,1H),7.73(dd,J＝8.0,3.3Hz,2H),7.67(dd,J＝8.5,4.3Hz,1H),7.32(d,J＝8.4Hz,1H),6.93(d,J＝8.5Hz,2H),5.11(dd,J＝12.8,5.4Hz,1H),4.71(m,1H),4.33(br,2H),3.86(m,3H),3.67(s,1H),3.50(m,1H),3.12(m,4H),3.02–2.72(m,5H),2.63–2.50(m,2H),2.23(m,2H),2.16–2.00(m,2H),1.93(m,3H),1.81(m,2H),1.59(m,4H),1.26(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ11.10(s,1H),9.52(s,1H),9.05(dd,J＝4.4,1.7Hz,1H),8.62(dd,J＝8.6,1.7Hz,1H),8.26(d,J ＝8.3Hz,1H),7.98(d,J＝7 .6Hz,1H),7.89(d,J＝9.2Hz,1H),7.73(dd,J＝8.0,3.3Hz,2H),7.67(dd,J＝8.5,4.3Hz,1H),7.32(d, J＝8.4Hz,1H),6.93(d,J ＝8.5Hz,2H),5.11(dd,J＝12.8,5.4Hz,1H),4.71(m,1H),4.33(br,2H),3.86(m,3H),3.67(s,1H),3.50 (m,1H),3.12(m,4H), 3.02–2.72(m,5H),2.63–2.50(m,2H),2.23(m,2H),2.16–2.00(m,2H),1.93(m,3H),1.81(m,2H),1.59( m,4H),1.26(m,2H).

实施例32化合物56的制备
Example 32 Preparation of Compound 56

向3-溴-2,4-二氟苯甲醛(56-1)(2g，9.0mmol，1.0eq.)于DMSO(20mL)中的溶液中添加80％ N2H4·H2O(2.2mL，36.0mmol，4.0eq.)。将反应混合物在120℃下搅拌5小时，使其冷却至室温，倒入H2O(80mL)中，然后过滤。将滤饼用H2O(20mL)洗涤并真空干燥，得到呈白色固体状的7-溴-6-氟-1H-吲唑(56-2)(1.1g，产率：56.8％)。MS:m/z＝216[M+H]+。To a solution of 3-bromo-2,4-difluorobenzaldehyde (56-1) (2 g, 9.0 mmol, 1.0 eq.) in DMSO (20 mL) was added 80% N 2 H 4 ·H 2 O (2.2 mL, 36.0 mmol, 4.0 eq.). The reaction mixture was stirred at 120° C. for 5 hours, allowed to cool to room temperature, poured into H 2 O (80 mL), and then filtered. The filter cake was washed with H 2 O (20 mL) and dried in vacuo to give 7-bromo-6-fluoro-1H-indazole (56-2) (1.1 g, yield: 56.8%) as a white solid. MS: m/z=216[M+H] + .

在0℃下，向7-溴-6-氟-1H-吲唑(56-2)(1.1g，5.1mmol，1.0eq.)和KOH(1.0g，17.8mmol，3.5eq.)于DMF(11mL)中的溶液中添加I2(1.9g，7.6mmol，1.5eq.)。将反应混合物在室温下搅拌2小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。合并 的有机层经Na2SO4干燥并真空浓缩，得到呈棕色油状的7-溴-6-氟-3-碘-1H-吲唑(56-3)(1.2g，粗物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝341[M+H]+。To a solution of 7-bromo-6-fluoro-1H-indazole (56-2) (1.1 g, 5.1 mmol, 1.0 eq.) and KOH (1.0 g, 17.8 mmol, 3.5 eq.) in DMF (11 mL) was added I 2 (1.9 g, 7.6 mmol, 1.5 eq.) at 0°C. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3). Combined The organic layer was dried over Na 2 SO 4 and concentrated in vacuo to give 7-bromo-6-fluoro-3-iodo-1H-indazole (56-3) (1.2 g, crude) as a brown oil. The crude material was used in the next step without purification. MS: m/z=341 [M+H] + .

在0℃下，向7-溴-6-氟-3-碘-1H-吲唑(56-3)(1.2g，3.5mmol，1.0eq.)和碘甲烷(749mg，5.2mmol，1.5eq.)于DMF(12mL)中的溶液中添加t-BuOK(796mg，7.1mmol，2.0eq.)。将反应混合物在室温下搅拌2小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将粗物质通过硅胶色谱用PE/EA(10:1)洗脱纯化，得到呈白色固体状的7-溴-6-氟-3-碘-1-甲基-1H-吲唑(56-4)(651mg，产率：52.2％)。MS:m/z＝355[M+H]+。At 0°C, t-BuOK (796 mg, 7.1 mmol, 2.0 eq.) was added to a solution of 7-bromo-6-fluoro-3-iodo-1H-indazole (56-3) (1.2 g, 3.5 mmol, 1.0 eq.) and iodomethane (749 mg, 5.2 mmol, 1.5 eq.) in DMF (12 mL). The reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The crude material was purified by silica gel chromatography eluting with PE/EA (10:1) to give 7-bromo-6-fluoro-3-iodo-1-methyl-1H-indazole (56-4) (651 mg, yield: 52.2%) as a white solid. MS: m/z=355[M+H] + .

向7-溴-6-氟-3-碘-1-甲基-1H-吲唑(56-4)(651mg，1.8mmol，1.0eq.)、2,6-双(苄氧基)-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊-2-基)吡啶(1.03g，2.4mmol，1.3eq.)和Cs2CO3(1.8g，5.5mmol，3.0eq.)于二恶烷(8mL)和H2O(2mL)中的溶液中添加Pd(dppf)Cl2(155mg，0.19mmol，0.1eq.)，将反应混合物在100℃下在N2下搅拌16小时。在反应完成后，将反应物冷却，通过硅藻土床过滤，用EA洗涤。分离有机层，经硫酸钠干燥并浓缩。将粗物质通过硅胶色谱用PE/EA(10:1)洗脱纯化，得到呈白色固体状的3-(2,6-双(苄氧基)吡啶-3-基)-7-溴-6-氟-1-甲基-1H-吲唑(56-5)(510mg，产率：53.7％)。MS:m/z＝519[M+H]+。To a solution of 7-bromo-6-fluoro-3-iodo-1-methyl-1H-indazole (56-4) (651 mg, 1.8 mmol, 1.0 eq.), 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine (1.03 g, 2.4 mmol, 1.3 eq.) and Cs2CO3 (1.8 g, 5.5 mmol, 3.0 eq.) in dioxane (8 mL) and H2O (2 mL) was added Pd( dppf ) Cl2 (155 mg, 0.19 mmol, 0.1 eq.) and the reaction mixture was stirred at 100°C under N2 for 16 h. After completion of the reaction, the reactant was cooled, filtered through a celite bed, washed with EA. The organic layer was separated, dried over sodium sulfate and concentrated. The crude material was purified by silica gel chromatography eluting with PE/EA (10:1) to give 3-(2,6-bis(benzyloxy)pyridin-3-yl)-7-bromo-6-fluoro-1-methyl-1H-indazole (56-5) (510 mg, yield: 53.7%) as a white solid. MS: m/z=519 [M+H] + .

向3-(2,6-双(苄氧基)吡啶-3-基)-7-溴-6-氟-1-甲基-1H-吲唑(56-5)(510mg，0.98mmol，1.0eq.)、t-Bu Xphos(83mg，0.2mmol，0.2eq.)和KOH(112mg，mmol，2.0eq.)于二恶烷(8mL)和H2O(2mL)中的溶液中添加Pd2(dba)3(92g，0.1mmol，0.1eq.)，将反应混合物在100℃下在N2下搅拌16小时。在反应完成后，将反应物冷却，通过硅藻土床过滤，用EA洗涤。分离有机层，经硫酸钠干燥并浓缩。将粗物质通过硅胶色谱用PE/EA(4:1)洗脱纯化，得到呈白色固体状的3-(2,6-双(苄氧基)吡啶-3-基)-6-氟-1-甲基-1H-吲唑-7-醇(56-6)(101mg，产率：22％)。MS:m/z＝457[M+H]+。To a solution of 3-(2,6-bis(benzyloxy)pyridin-3-yl)-7-bromo-6-fluoro-1-methyl-1H-indazole (56-5) (510 mg, 0.98 mmol, 1.0 eq.), t-BuXphos (83 mg, 0.2 mmol, 0.2 eq.) and KOH (112 mg, mmol, 2.0 eq.) in dioxane (8 mL) and H 2 O (2 mL) was added Pd 2 (dba) 3 (92 g, 0.1 mmol, 0.1 eq.) and the reaction mixture was stirred at 100 °C under N 2 for 16 h. After completion of the reaction, the reactant was cooled, filtered through a celite bed, and washed with EA. The organic layer was separated, dried over sodium sulfate and concentrated. The crude material was purified by silica gel chromatography eluting with PE/EA (4:1) to give 3-(2,6-bis(benzyloxy)pyridin-3-yl)-6-fluoro-1-methyl-1H-indazol-7-ol (56-6) (101 mg, yield: 22%) as a white solid. MS: m/z=457 [M+H] + .

向3-(2,6-双(苄氧基)吡啶-3-基)-6-氟-1-甲基-1H-吲唑-7-醇(56-6)(101mg，0.22mmol，1.0eq.)和(R)-3-((甲苯磺酰基氧基)甲基)吡咯烷-1-甲酸叔丁酯(79mg，0.22mmol，1.0eq.)于DMF(2mL)中的溶液中添加K2CO3(91mg，0.66mmol，3.0eq.)。将反应混合物在80℃下搅拌16小时。在反应完成后，将混合物用H2O(20mL)稀释并用EA(20mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将残余物通过硅胶柱色谱(PE/EA＝10:1)纯化，得到呈白色固体状的(R)-3-(((3-(2,6-双(苄氧基)吡啶-3-基)-6-氟-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(56-7)(121mg，产率：85.7％)。MS:m/z＝639[M+H]+。To a solution of 3-(2,6-bis(benzyloxy)pyridin-3-yl)-6-fluoro-1-methyl-1H-indazol-7-ol (56-6) (101 mg, 0.22 mmol, 1.0 eq.) and (R)-tert-butyl 3-((tosyloxy)methyl)pyrrolidine-1-carboxylate (79 mg, 0.22 mmol, 1.0 eq.) in DMF (2 mL) was added K 2 CO 3 (91 mg, 0.66 mmol, 3.0 eq.). The reaction mixture was stirred at 80° C. for 16 h. After the reaction was completed, the mixture was diluted with H 2 O (20 mL) and extracted with EA (20 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The residue was purified by silica gel column chromatography (PE/EA=10:1) to give (R)-tert-butyl 3-(((3-(2,6-bis(benzyloxy)pyridin-3-yl)-6-fluoro-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (56-7) (121 mg, yield: 85.7%) as a white solid. MS: m/z=639 [M+H] + .

向(R)-3-(((3-(2,6-双(苄氧基)吡啶-3-基)-6-氟-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(56-7)(121mg，0.18mmol，1.0eq.)和10％ Pd/C(24mg，20重量％)于EtOH(3mL)中的溶液中添加(Boc)2O(41mg，0.18mmol，1.0eq.)，将反应混合物在室温下在H2气氛(50Psi)下搅拌16小时。在反应完成后，过滤混合物并真空浓缩，得到呈棕色固体状的(3R)-3-(((3-(2,6-二氧代哌啶-3-基)-6-氟-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(56-8)(81mg，粗 物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝361[M+H-100]+。To a solution of (R)-tert-butyl 3-(((3-(2,6-bis(benzyloxy)pyridin-3-yl)-6-fluoro-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (56-7) (121 mg, 0.18 mmol, 1.0 eq.) and 10% Pd/C (24 mg, 20 wt%) in EtOH (3 mL) was added (Boc) 2O (41 mg, 0.18 mmol, 1.0 eq.) and the reaction mixture was stirred at room temperature under H2 atmosphere (50 Psi) for 16 h. After the reaction was complete, the mixture was filtered and concentrated in vacuo to give tert-butyl (3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-6-fluoro-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (56-8) (81 mg, crude) as a brown solid. The crude material was used in the next step without purification. MS: m/z = 361 [M+H-100] + .

向(3R)-3-(((3-(2,6-二氧代哌啶-3-基)-6-氟-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(56-8)(81mg，0.17mmol，1.0eq.)于EA(2mL)中的溶液中添加4M HCl/EA(2mL)，将反应混合物在室温下搅拌6小时。在反应完成后，真空浓缩混合物，得到呈绿色固体状的3-(6-氟-1-甲基-7-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2,6-二酮盐酸盐(56-9)(55mg，粗物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝361[M+H]+。To a solution of tert-butyl (3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-6-fluoro-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (56-8) (81 mg, 0.17 mmol, 1.0 eq.) in EA (2 mL) was added 4M HCl/EA (2 mL), and the reaction mixture was stirred at room temperature for 6 hours. After the reaction was complete, the mixture was concentrated in vacuo to give 3-(6-fluoro-1-methyl-7-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione hydrochloride (56-9) (55 mg, crude) as a green solid. The crude material was used in the next step without purification. MS: m/z=361 [M+H] + .

向3-(6-氟-1-甲基-7-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2,6-二酮盐酸盐(56-9)(55mg，0.15mmol，1.0eq.)和N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(72mg，0.15mmol，1.0eq.)于DMF(2mL)中的溶液中添加TEA(61mg，0.61mmol，4.0eq.)，将反应混合物在室温下搅拌2小时，然后向混合物中添加NaBH(OAc)3(48mg，0.225mmol，1.5eq.)并在室温下搅拌2小时。在反应完成后，将混合物通过反相色谱用MeCN:H2O＝56％洗脱纯化，得到呈白色固体状的N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)-6-氟-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(56)(31.9mg，产率：25.5％)。MS:m/z＝827[M+H]+。To a solution of 3-(6-fluoro-1-methyl-7-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione hydrochloride (56-9) (55 mg, 0.15 mmol, 1.0 eq.) and N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (72 mg, 0.15 mmol, 1.0 eq.) in DMF (2 mL) was added TEA (61 mg, 0.61 mmol, 4.0 eq.), the reaction mixture was stirred at room temperature for 2 hours, and then NaBH(OAc) 3 (48 mg, 0.225 mmol, 1.5 eq.) was added to the mixture and stirred at room temperature for 2 hours. After the reaction was completed, the mixture was purified by reverse phase chromatography eluting with MeCN:H 2 O=56% to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-6-fluoro-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (56) (31.9 mg, yield: 25.5%) as a white solid. MS: m/z=827[M+H] + .

1H NMR(400MHz,DMSO-d6)δ11.12(s,1H),9.07(dd,J＝4.2,1.8Hz,1H),8.64(dd,J＝8.5,1.8Hz,1H),8.29(d,J＝8.3Hz,1H),7.89(d,J＝8.3Hz,1H),7.75–7.67(m,2H),7.52–7.46(m,2H),7.39(d,J＝8.2Hz,1H),7.34(d,J＝8.4Hz,1H),6.84–6.75(m,2H),5.13(dd,J＝12.8,5.4Hz,1H),4.79–4.70(m,1H),4.24–4.17(m,1H),3.94–3.84(m,4H),3.19–3.09(m,4H),3.03–2.92(m,2H),2.89–2.76(m,4H),2.69–2.58(m,2H),2.25–2.19(m,2H),2.08–1.92(m,7H),1.85–1.78(m,3H),1.71–1.54(m,4H). 1 H NMR (400MHz, DMSO-d 6 )δ11.12(s,1H),9.07(dd,J＝4.2,1.8Hz,1H),8.64(dd,J＝8.5,1.8Hz,1H),8.29(d,J＝8.3Hz,1H), 7.89(d,J＝8.3Hz, 1H),7.75–7.67(m,2H),7.52–7.46(m,2H),7.39(d,J＝8.2Hz,1H),7.34(d,J＝8.4Hz,1H),6.84–6.75(m ,2H),5.13(dd ,J＝12.8,5.4Hz,1H),4.79–4.70(m,1H),4.24–4.17(m,1H),3.94–3.84(m,4H),3.19–3.09(m,4H),3.03–2.92 (m,2H) ,2.89–2.76(m,4H),2.69–2.58(m,2H),2.25–2.19(m,2H),2.08–1.92(m,7H),1.85–1.78(m,3H),1.71–1.54( m,4H).

实施例33化合物57的制备
Example 33 Preparation of Compound 57

向2,4-二氟苯酚(3g，23.0mmol，1.0eq.)和(R)-3-((甲苯磺酰基氧基)甲基)吡咯烷-1-甲酸叔丁酯(8.2g，23.0mmol，1.0eq.)于DMF(30mL)中的溶液中添加K2CO3(9.5g，69.1mmol，3.0eq.)。将反应混合物在80℃下搅拌16小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将残余物通过硅胶柱色谱 (PE/EA＝10:1)纯化，得到呈无色油状的(R)-3-((2,4-二氟苯氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-1)(4.8g，产率：66％)。MS:m/z＝314[M+H]+。To a solution of 2,4-difluorophenol (3 g, 23.0 mmol, 1.0 eq.) and (R)-tert-butyl 3-((tosyloxy)methyl)pyrrolidine-1-carboxylate (8.2 g, 23.0 mmol, 1.0 eq.) in DMF (30 mL) was added K 2 CO 3 (9.5 g, 69.1 mmol, 3.0 eq.). The reaction mixture was stirred at 80° C. for 16 hours. After the reaction was completed, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The residue was purified by silica gel column chromatography After purification by HPLC (PE/EA=10:1), (R)-tert-butyl 3-((2,4-difluorophenoxy)methyl)pyrrolidine-1-carboxylate (57-1) (4.8 g, yield: 66%) was obtained as a colorless oil. MS: m/z=314 [M+H] + .

在-78℃下，向(R)-3-((2,4-二氟苯氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-1)(4.8g，15.3mmol，1.0eq.)于THF(50mL)中的溶液中添加n-BuLi(6.7mL，2.5M)。将反应混合物在-78℃下搅拌1小时。逐滴添加DMF(1.7g，22.9mmol，1.5eq.)并将混合物在-78℃下搅拌1小时。在反应完成后，在相同温度下，缓慢地添加H2O(50mL)。使混合物升温至室温并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩，得到呈无色油状的(R)-3-((2,4-二氟-3-甲酰基苯氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-2)(4.8g，粗物质)。MS:m/z＝342[M+H]+。To a solution of (R)-tert-butyl 3-((2,4-difluorophenoxy)methyl)pyrrolidine-1-carboxylate (57-1) (4.8 g, 15.3 mmol, 1.0 eq.) in THF (50 mL) was added n-BuLi (6.7 mL, 2.5 M) at -78 °C. The reaction mixture was stirred at -78 °C for 1 hour. DMF (1.7 g, 22.9 mmol, 1.5 eq.) was added dropwise and the mixture was stirred at -78 °C for 1 hour. After the reaction was completed, H 2 O (50 mL) was slowly added at the same temperature. The mixture was allowed to warm to room temperature and extracted with EA (50 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated to give (R)-tert-butyl 3-((2,4-difluoro-3-formylphenoxy)methyl)pyrrolidine-1-carboxylate (57-2) (4.8 g, crude) as a colorless oil. MS: m/z = 342 [M+H] + .

向(R)-3-((2,4-二氟-3-甲酰基苯氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-2)(2g，5.8mmol，1.0eq.)于DMSO(20mL)中的溶液中添加水合肼80％(1.5g，23.4mmol，4.0eq.)。将反应混合物在120℃下搅拌16小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将残余物通过硅胶柱色谱(PE/EA＝10:1)纯化，得到呈无色油状的(R)-3-(((4-氟-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-3)(705mg，产率：36％)。MS:m/z＝336[M+H]+。To a solution of (R)-tert-butyl 3-((2,4-difluoro-3-formylphenoxy)methyl)pyrrolidine-1-carboxylate (57-2) (2 g, 5.8 mmol, 1.0 eq.) in DMSO (20 mL) was added hydrazine hydrate 80% (1.5 g, 23.4 mmol, 4.0 eq.). The reaction mixture was stirred at 120° C. for 16 hours. After the reaction was completed, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The residue was purified by silica gel column chromatography (PE/EA=10:1) to give (R)-tert-butyl 3-(((4-fluoro-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-3) (705 mg, yield: 36%) as a colorless oil. MS: m/z = 336 [M+H] + .

在0℃下，向(R)-3-(((4-氟-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-3)(705mg，2.1mmol，1.0eq.)和KOH(412mg，7.3mmol，3.5eq.)于DMF(7mL)中的溶液中添加I2(800mg，3.1mmol，1.5eq.)。将反应混合物在室温下搅拌2小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并真空浓缩，得到呈棕色油状的(R)-3-(((4-氟-3-碘-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-4)(951mg，粗物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝462[M+H]+。To a solution of (R)-tert-butyl 3-(((4-fluoro-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-3) (705 mg, 2.1 mmol, 1.0 eq.) and KOH (412 mg, 7.3 mmol, 3.5 eq.) in DMF (7 mL) at 0°C was added I2 (800 mg, 3.1 mmol, 1.5 eq.). The reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was diluted with H2O (50 mL ) and extracted with EA (50 mL x 3). The combined organic layers were dried over Na2SO4 and concentrated in vacuo to give (R)-tert-butyl 3-(((4-fluoro-3-iodo-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-4) (951 mg, crude) as a brown oil. The crude material was used in the next step without purification. MS: m/z = 462 [M+H] + .

在0℃下，向(R)-3-(((4-氟-3-碘-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-4)(951mg，2.06mmol，1.0eq.)和碘甲烷(438mg，3.09mmol，1.5eq.)于DMF(10mL)中的溶液中添加t-BuOK(462mg，4.12mmol，2.0eq.)。将反应混合物在室温下搅拌2小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。将合并的有机层经Na2SO4干燥并真空浓缩，得到呈棕色油状的(R)-3-(((4-氟-3-碘-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-5)(902mg，粗物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝476[M+H]+。To a solution of (R)-tert-butyl 3-(((4-fluoro-3-iodo-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-4) (951 mg, 2.06 mmol, 1.0 eq.) and iodomethane (438 mg, 3.09 mmol, 1.5 eq.) in DMF (10 mL) at 0°C was added t-BuOK (462 mg, 4.12 mmol, 2.0 eq.). The reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the mixture was diluted with H2O (50 mL) and extracted with EA (50 mL×3). The combined organic layers were dried over Na2SO4 and concentrated in vacuo to give (R)-tert-butyl 3 -(((4-fluoro-3-iodo-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-5) (902 mg, crude) as a brown oil. The crude material was used in the next step without purification. MS: m/z = 476 [M+H] + .

向(R)-3-(((4-氟-3-碘-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-5)(902mg，1.9mmol，1.0eq.)、2,6-双(苄氧基)-3-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊-2-基)吡啶(57-3)(1.03g，2.46mmol，1.3eq.)和Cs2CO3(1.8g，5.7mmol，3.0eq.)于二恶烷(12mL)和H2O(3mL)中的溶液中添加Pd(dppf)Cl2(155mg，0.19mmol，0.1eq.)，将反应混合物在100℃下在N2下搅拌16小时。在反应完成后，将反应物冷却，通过硅藻土床过滤，用EA洗涤。分离有机层，经硫酸钠干燥并浓缩。将粗物质通过硅胶色谱用PE/EA(10:1)洗脱纯化，得到呈棕色油状的(R)-3-(((3-(2,6-双(苄氧基)吡啶-3-基)-4-氟-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-6)(851mg，产率：70％)。MS:m/z＝639[M+H]+。 To a solution of (R)-tert-butyl 3-(((4-fluoro-3-iodo-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-5) (902 mg, 1.9 mmol, 1.0 eq.), 2,6-bis(benzyloxy)-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) pyridine (57-3) (1.03 g, 2.46 mmol, 1.3 eq.) and Cs2CO3 (1.8 g, 5.7 mmol, 3.0 eq.) in dioxane (12 mL) and H2O (3 mL) was added Pd(dppf) Cl2 (155 mg, 0.19 mmol, 0.1 eq.) and the reaction mixture was stirred at 100 °C under N2 for 16 h. After the reaction was completed, the reactant was cooled, filtered through a celite bed, and washed with EA. The organic layer was separated, dried over sodium sulfate and concentrated. The crude material was purified by silica gel chromatography eluting with PE/EA (10:1) to give (R)-3-(((3-(2,6-bis(benzyloxy)pyridin-3-yl)-4-fluoro-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylic acid tert-butyl ester (57-6) (851 mg, yield: 70%) as a brown oil. MS: m/z=639[M+H] + .

向(R)-3-(((3-(2,6-双(苄氧基)吡啶-3-基)-4-氟-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-6)(851mg，1.3mmol，1.0eq.)和10％ Pd/C(170mg，20重量％)于EtOH(8mL)中的溶液中添加(Boc)2O(291mg，1.3mmol，1.0eq.)，将反应混合物在室温下在H2气氛(50Psi)下搅拌16小时。在反应完成后，过滤混合物并真空浓缩，得到呈棕色油状的(3R)-3-(((3-(2,6-二氧代哌啶-3-基)-4-氟-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-7)(604mg，粗物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝361[M+H-100]+。To a solution of (R)-tert-butyl 3-(((3-(2,6-bis(benzyloxy)pyridin-3-yl)-4-fluoro-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-6) (851 mg, 1.3 mmol, 1.0 eq.) and 10% Pd/C (170 mg, 20 wt%) in EtOH (8 mL) was added (Boc) 2O (291 mg, 1.3 mmol, 1.0 eq.) and the reaction mixture was stirred at room temperature under H2 atmosphere (50 Psi) for 16 h. After the reaction was complete, the mixture was filtered and concentrated in vacuo to give tert-butyl (3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-7) (604 mg, crude) as a brown oil. The crude material was used in the next step without purification. MS: m/z=361 [M+H-100] + .

向(3R)-3-(((3-(2,6-二氧代哌啶-3-基)-4-氟-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(57-7)(604mg，1.3mmol，1.0eq.)于EA(2mL)中的溶液中添加4M HCl/EA(3mL)，将反应混合物在室温下搅拌6小时。在反应完成后，真空浓缩混合物，得到呈绿色固体状的3-(4-氟-1-甲基-5-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2,6-二酮盐酸盐(57-8)(352mg，粗物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝361[M+H]+。To a solution of tert-butyl (3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidine-1-carboxylate (57-7) (604 mg, 1.3 mmol, 1.0 eq.) in EA (2 mL) was added 4M HCl/EA (3 mL), and the reaction mixture was stirred at room temperature for 6 hours. After the reaction was complete, the mixture was concentrated in vacuo to give 3-(4-fluoro-1-methyl-5-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione hydrochloride (57-8) (352 mg, crude) as a green solid. The crude material was used in the next step without purification. MS: m/z=361 [M+H] + .

向3-(4-氟-1-甲基-5-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2,6-二酮盐酸盐(57-8)(100mg，0.25mmol，1.0eq.)和N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-甲酰基哌啶-1-基)苯甲酰胺(5-5)(121mg，0.25mmol，1.0eq.)于DMF(2mL)中的溶液中添加TEA(106mg，1.05mmol，4.0eq.)，将反应混合物在室温下搅拌4小时，然后向混合物中添加NaBH(OAc)3(79mg，0.37mmol，1.5eq.)并在室温下搅拌4小时。在反应完成后，将混合物通过反相色谱用MeCN:H2O＝56％洗脱纯化，得到呈白色固体状的N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)-4-氟-1-甲基-1H-吲唑-5-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)苯甲酰胺(57)(47.9mg，产率：23％)。MS:m/z＝827[M+H]+。To a solution of 3-(4-fluoro-1-methyl-5-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione hydrochloride (57-8) (100 mg, 0.25 mmol, 1.0 eq.) and N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-formylpiperidin-1-yl)benzamide (5-5) (121 mg, 0.25 mmol, 1.0 eq.) in DMF (2 mL) was added TEA (106 mg, 1.05 mmol, 4.0 eq.), the reaction mixture was stirred at room temperature for 4 hours, and then NaBH(OAc) 3 (79 mg, 0.37 mmol, 1.5 eq.) was added to the mixture and stirred at room temperature for 4 hours. After the reaction was completed, the mixture was purified by reverse phase chromatography eluting with MeCN:H 2 O=56% to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-4-fluoro-1-methyl-1H-indazol-5-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)benzamide (57) (47.9 mg, yield: 23%) as a white solid. MS: m/z=827[M+H] + .

1H NMR(400MHz,DMSO-d6)δ10.88(s,1H),9.05(dd,J＝4.3,1.7Hz,1H),8.61(dd,J＝8.5,1.7Hz,1H),8.26(d,J＝8.3Hz,1H),7.98(d,J＝7.6Hz,1H),7.73(d,J＝8.4Hz,2H),7.66(dd,J＝8.5,4.2Hz,1H),7.32(d,J＝8.4Hz,1H),6.98–6.89(m,2H),6.80–6.70(m,2H),4.71(brs,1H),4.28(dd,J＝11.4,5.2Hz,1H),4.23–4.04(m,5H),3.85(d,J＝12.7Hz,4H),3.71(brs,1H),3.61–3.51(m,1H),3.45–3.32(m,1H),3.23–3.17(m,3H),3.08–2.95(m,1H),2.93–2.83(m,1H),2.82–2.68(m,3H),2.61–2.51(m,1H),2.32–2.18(m,3H),2.17–2.05(m,1H),2.03–1.87(m,3H),1.86–1.77(m,2H),1.71–1.52(m,4H),1.35–1.18(m,2H). 1 H NMR (400MHz, DMSO-d6) δ10.88(s,1H),9.05(dd,J＝4.3,1.7Hz,1H),8.61(dd,J＝8.5,1.7Hz,1H),8.26(d,J ＝8.3Hz,1H),7.98(d,J＝7.6Hz,1H),7.73(d,J ＝8.4Hz,2H),7.66(dd,J＝8.5,4.2Hz,1H),7.32(d,J＝8.4Hz,1H),6.98–6.89 (m,2H),6.80–6.70(m,2H),4.71(brs,1H),4.28(dd,J＝11.4,5.2Hz,1H),4.2 3–4.04(m,5H),3.85(d,J＝12.7Hz,4H),3.71(brs,1H),3.61–3.51(m,1H),3 .45–3.32(m,1H),3.23–3.17(m,3H),3.08–2.95(m,1H),2.93–2.83(m,1H),2 .82–2.68(m,3H),2.61–2.51(m,1H),2.32–2.18(m,3H),2.17–2.05(m,1H), 2.03–1.87(m,3H),1.86–1.77(m,2H),1.71–1.52(m,4H),1.35–1.18(m,2H).

实施例34化合物58和58A的制备
Example 34 Preparation of Compounds 58 and 58A

向4-((2R,4R)-4-(甲氧基羰基)-2-甲基哌啶-1-基)苯甲酸(80mg，0.28mmol，1.0eq.)于DMF(1mL)和DIEA(149mg，1.15mmol，4.0eq.)中的溶液中添加HATU(164mg，0.43mmol，1.5eq.)。将反应混合物在室温下搅拌20分钟。添加5-(((1r,4r)-4-氨基环己基)氧基)喹啉-8-甲腈盐酸盐(87mg，0.31mmol，1.0eq.)。将反应物在室温下搅拌2小时。在反应完成后，将混合物在H2O(20mL)与EA(20mL)之间分配，并用EA(3×20mL)萃取水层。将有机层干燥并蒸发。将残余物通过硅胶色谱用纯EA洗脱纯化，得到呈黄色固体状的(2R,4R)-1-(4-(((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)氨基甲酰基)苯基)-2-甲基哌啶-4-甲酸甲酯(58-2)(51mg，33％产率)。MS:m/z＝527[M+H]+。To a solution of 4-((2R,4R)-4-(methoxycarbonyl)-2-methylpiperidin-1-yl)benzoic acid (80 mg, 0.28 mmol, 1.0 eq.) in DMF (1 mL) and DIEA (149 mg, 1.15 mmol, 4.0 eq.) was added HATU (164 mg, 0.43 mmol, 1.5 eq.). The reaction mixture was stirred at room temperature for 20 min. 5-(((1r,4r)-4-aminocyclohexyl)oxy)quinoline-8-carbonitrile hydrochloride (87 mg, 0.31 mmol, 1.0 eq.) was added. The reactants were stirred at room temperature for 2 h. After the reaction was complete, the mixture was partitioned between H 2 O (20 mL) and EA (20 mL), and the aqueous layer was extracted with EA (3×20 mL). The organic layer was dried and evaporated. The residue was purified by silica gel chromatography eluting with pure EA to give methyl (2R,4R)-1-(4-(((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)carbamoyl)phenyl)-2-methylpiperidine-4-carboxylate (58-2) (51 mg, 33% yield) as a yellow solid. MS: m/z = 527 [M+H] + .

向(2R,4R)-1-(4-(((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)氨基甲酰基)苯基)-2-甲基哌啶-4-甲酸甲酯(58-2)(51mg，0.09mmol，1.0eq.)于THF(1mL)中的溶液中添加LiBH4(21mg，0.96mmol，10.0eq.)。将反应混合物在室温下搅拌16小时。在反应完成后，将混合物在H2O(20mL)与EA(20mL)之间分配，并用EA(3×20mL)萃取水层。将有机层干燥并蒸发，得到呈黄色固体状的N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4R)-4-(羟甲基)-2-甲基哌啶-1-基)苯甲酰胺(58-3)(31mg，粗物质)。MS:m/z＝499[M+H]+。To a solution of methyl (2R,4R)-1-(4-(((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)carbamoyl)phenyl)-2-methylpiperidine-4-carboxylate (58-2) (51 mg, 0.09 mmol, 1.0 eq.) in THF (1 mL) was added LiBH4 (21 mg, 0.96 mmol, 10.0 eq.). The reaction mixture was stirred at room temperature for 16 h. After completion of the reaction, the mixture was partitioned between H2O (20 mL) and EA (20 mL), and the aqueous layer was extracted with EA (3 x 20 mL). The organic layer was dried and evaporated to give N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4R)-4-(hydroxymethyl)-2-methylpiperidin-1-yl)benzamide (58-3) (31 mg, crude) as a yellow solid. MS: m/z = 499 [M+H] + .

向N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4R)-4-(羟甲基)-2-甲基哌啶-1-基)苯甲酰胺(58-3)(31mg，0.06mmol，1.0eq.)和戴斯-马丁试剂(Dess-Martin reagent)(29mg，0.06mmol，1.1eq.)于DCM(2mL)中的溶液中。将反应混合物在室温下搅拌3小时。在反应完成后，将混合物在H2O(20mL)与DCM(20mL)之间分配，并用DCM(3×20mL)萃取水层。将有机层干燥并蒸发。将残余物通过反相色谱(TFA缓冲液)纯化，得到呈白色固体状的N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4R)-4-甲酰基-2-甲基哌啶-1-基)苯甲酰胺(58-4)(19mg，61％产率)。MS:m/z＝497[M+H]+。To a solution of N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4R)-4-(hydroxymethyl)-2-methylpiperidin-1-yl)benzamide (58-3) (31 mg, 0.06 mmol, 1.0 eq.) and Dess-Martin reagent (29 mg, 0.06 mmol, 1.1 eq.) in DCM (2 mL). The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was partitioned between H 2 O (20 mL) and DCM (20 mL), and the aqueous layer was extracted with DCM (3×20 mL). The organic layer was dried and evaporated. The residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4R)-4-formyl-2-methylpiperidin-1-yl)benzamide (58-4) (19 mg, 61% yield) as a white solid. MS: m/z = 497 [M+H] + .

N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4S)-4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)苯甲酰胺(58)N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4S)-4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidin-1-yl)methyl)-2-methylpiperidin-1-yl)benzamide (58)

N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4R)-4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)苯甲酰胺(58A)N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4R)-4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidin-1-yl)methyl)-2-methylpiperidin-1-yl)benzamide (58A)

向N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4R)-4-甲酰基-2-甲基哌啶-1-基)苯甲酰胺(58-4)(19mg，0.038mmol，1.0eq.)和3-(1-甲基-7-(((R)-吡咯烷-3-基)甲氧基)-1H-吲唑-3-基)哌啶-2,6-二酮盐酸盐(15mg，0.1mmol，1.0eq.)于DMF(1mL)中的溶液中添加TEA(19mg，0.19mmol，5.0eq.)。将反应物在室温下搅拌2小时，然后添加NaBH(OAc)3(12mg，0.057mmol，1.5eq.)。将反应物在室温下搅拌2小时。在反应完成后，将残余物通过反相色谱(TFA缓冲液)纯化，得到呈白色固体状的N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4S)-4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)-1-甲基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)苯甲酰胺(58)(3.6mg，11.5％产率，Rf＝0.6，DCM:MeOH＝10:1)和呈白色固体状的N-((1r,4R)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-((2R,4R)-4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)-1-甲 基-1H-吲唑-7-基)氧基)甲基)吡咯烷-1-基)甲基)-2-甲基哌啶-1-基)苯甲酰胺(58A)(4mg，12％产率，Rf＝0.5，DCM:MeOH＝10:1)。MS:m/z＝823[M+H]+。To a solution of N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4R)-4-formyl-2-methylpiperidin-1-yl)benzamide (58-4) (19 mg, 0.038 mmol, 1.0 eq.) and 3-(1-methyl-7-(((R)-pyrrolidin-3-yl)methoxy)-1H-indazol-3-yl)piperidine-2,6-dione hydrochloride (15 mg, 0.1 mmol, 1.0 eq.) in DMF (1 mL) was added TEA (19 mg, 0.19 mmol, 5.0 eq.). The reaction was stirred at room temperature for 2 hours, then NaBH(OAc) 3 (12 mg, 0.057 mmol, 1.5 eq.) was added. The reaction was stirred at room temperature for 2 hours. After the reaction was complete, the residue was purified by reverse phase chromatography (TFA buffer) to give N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4S)-4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-1-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidin-1-yl)methyl as a white solid. )-2-methylpiperidin-1-yl)benzamide (58) (3.6 mg, 11.5% yield, Rf = 0.6, DCM:MeOH = 10:1) and N-((1r,4R)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-((2R,4R)-4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)-1-methyl)- 1-(4-(4-(4-(4-methyl-1H-indazol-7-yl)oxy)methyl)pyrrolidin-1-yl)methyl)-2-methylpiperidin-1-yl)benzamide (58A) (4 mg, 12% yield, Rf=0.5, DCM:MeOH=10:1). MS: m/z=823 [M+H] + .

化合物58：1H NMR(400MHz,DMSO-d6)δ10.89(s,1H),9.08(dd,J＝4.3,1.8Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8.29(d,J＝8.3Hz,1H),7.98(d,J＝7.6Hz,1H),7.75(d,J＝8.3Hz,2H),7.71–7.68(m,1H),7.36(d,J＝8.5Hz,1H),7.27(d,J＝8.2Hz,1H),7.02(t,J＝7.8Hz,1H),6.92(d,J＝8.4Hz,2H),6.86(d,J＝7.6Hz,1H),4.73(brs,1H),4.43–4.25(m,3H),4.22–4.13(m,4H),3.94–3.83(m,2H),3.78–3.67(m,1H),3.63–3.57(m,2H),3.21–3.04(m,4H),2.99–2.86(m,2H),2.73–2.60(m,2H),2.35–2.31(m,2H),2.28–2.23(m,2H),2.21–2.14(m,2H),1.98–1.91(m,2H),1.90–1.80(m,1H),1.79–1.73(m,1H),1.69–1.56(m,4H),1.27–1.18(m,2H),1.04(d,J＝6.5Hz,3H).Compound 58: 1 H NMR (400 MHz, DMSO-d 6 )δ10.89(s,1H),9.08(dd,J＝4.3,1.8Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8. 29(d,J＝8.3Hz,1H),7.98(d,J＝7.6Hz,1H),7.75(d,J＝8.3Hz,2H),7.71–7.68( m,1H),7.36(d,J＝8.5Hz,1H),7.27(d,J＝8.2Hz,1H),7.02(t,J＝7.8Hz,1H),6. 92(d,J＝8.4Hz,2H),6.86(d,J＝7.6Hz,1H),4.73(brs,1H),4.43–4.25(m,3H), 4.22–4.13(m,4H),3.94–3.83(m,2H),3.78–3.67(m,1H),3.63–3.57(m,2H), 3.21–3.04(m,4H),2.99–2.86(m,2H),2.73–2.60(m,2H),2.35–2.31(m,2H),2 .28–2.23(m,2H),2.21–2.14(m,2H),1.98–1.91(m,2H),1.90–1.80(m,1H),1. 79–1.73(m,1H),1.69–1.56(m,4H),1.27–1.18(m,2H),1.04(d,J＝6.5Hz,3H).

化合物58A：1H NMR(400MHz,DMSO-d6)δ10.88(s,1H),9.07(d,J＝2.8Hz,1H),8.65(d,J＝8.3Hz,1H),8.29(d,J＝8.3Hz,1H),8.11(d,J＝7.6Hz,1H),7.76(d,J＝8.5Hz,2H),7.69(dd,J＝8.5,4.2Hz,1H),7.36(d,J＝8.6Hz,1H),7.22(d,J＝8.0Hz,1H),7.04–6.95(m,3H),6.84(d,J＝7.4Hz,1H),4.77–4.72(m,1H),4.32(dd,J＝9.9,5.1Hz,1H),4.19(s,3H),4.16–4.11(m,2H),4.07–4.02(m,2H),3.95–3.81(m,3H),2.97–2.90(m,1H),2.40–2.22(m,6H),2.21–2.08(m,2H),2.07–1.84(m,6H),1.70–1.57(m,6H),1.31–1.19(m,4H),0.96(d,J＝5.9Hz,3H).Compound 58A: 1 H NMR (400MHz, DMSO-d 6 )δ10.88(s,1H),9.07(d,J＝2.8Hz,1H),8.65(d,J＝8.3Hz,1H),8.29(d,J＝8.3Hz,1H),8.11(d, J＝7.6Hz,1H),7.76(d,J＝8.5Hz, 2H),7.69(dd,J＝8.5,4.2Hz,1H),7.36(d,J＝8.6Hz,1H),7.22(d,J＝8.0Hz,1H),7.04–6.95(m,3H), 6.84(d,J＝7.4Hz,1H),4.77 –4.72(m,1H),4.32(dd,J＝9.9,5.1Hz,1H),4.19(s,3H),4.16–4.11(m,2H),4.07–4.02(m,2H),3.95–3.81 (m,3H),2.97–2.90( m,1H),2.40–2.22(m,6H),2.21–2.08(m,2H),2.07–1.84(m,6H),1.70–1.57(m,6H),1.31–1.19(m,4H), 0.96(d,J＝5.9Hz,3H).

实施例35化合物67和67A参照化合物58和58A的制备合成。Example 35 Compounds 67 and 67A were synthesized by referring to the preparation of compounds 58 and 58A.

实施例36化合物86的制备
Example 36 Preparation of Compound 86

在0℃下，向1-(3-溴-2-羟基苯基)乙-1-酮(5g，25.0mmol，1.0eq.)和碳酸二乙酯(11.8g，100.0mmol，4.0eq.)于甲苯(50mL)中的溶液中添加60％ NaH(4g，100.0mmol，4.0eq.)。将反应混合物在120℃下搅拌16小时。在反应完成后，将残余物与冰-水(200mL)混合，用HCl水溶液(3M)酸化至pH 2-3并用EA(100mL×3)萃取。合并的有机层经Na2SO4干燥并真空浓缩，得到呈黄色固体状的3-(3-溴-2-羟基苯基)-3-氧代丙酸乙酯(86-1)(5g，粗物质)。MS:m/z＝288[M+H]+。To a solution of 1-(3-bromo-2-hydroxyphenyl)ethan-1-one (5 g, 25.0 mmol, 1.0 eq.) and diethyl carbonate (11.8 g, 100.0 mmol, 4.0 eq.) in toluene (50 mL) was added 60% NaH (4 g, 100.0 mmol, 4.0 eq.) at 0°C. The reaction mixture was stirred at 120°C for 16 hours. After the reaction was completed, the residue was mixed with ice-water (200 mL), acidified to pH 2-3 with aqueous HCl (3 M) and extracted with EA (100 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated in vacuo to give ethyl 3-(3-bromo-2-hydroxyphenyl)-3-oxopropanoate (86-1) (5 g, crude material) as a yellow solid. MS: m/z=288[M+H] + .

向3-(3-溴-2-羟基苯基)-3-氧代丙酸乙酯(86-1)(5g，粗物质)于DMF(50mL)中的溶液中添加K2CO3(7.2g，52.2mmol，3.0eq.)。将反应混合物在120℃下搅拌4小时。在反应完成后，将残余物与冰-水(200mL)混合，用HCl水溶液(3M)酸化至pH 2-3并用EA(100mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将残余物通过逆相纯化，得到呈黄色固体状的8- 溴-4-羟基-2H-苯并吡喃-2-酮(86-2)(2.6g，43％两步骤产率)。MS:m/z＝242[M+H]+。To a solution of ethyl 3-(3-bromo-2-hydroxyphenyl)-3-oxopropanoate (86-1) (5 g, crude material) in DMF (50 mL) was added K 2 CO 3 (7.2 g, 52.2 mmol, 3.0 eq.). The reaction mixture was stirred at 120° C. for 4 hours. After the reaction was completed, the residue was mixed with ice-water (200 mL), acidified to pH 2-3 with aqueous HCl (3 M) and extracted with EA (100 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The residue was purified by reverse phase to give 8- Bromo-4-hydroxy-2H-benzopyran-2-one (86-2) (2.6 g, 43% yield in two steps). MS: m/z=242 [M+H] + .

向8-溴-4-羟基-2H-苯并吡喃-2-酮(86-2)(2.6g，10.7mmol，1.0eq.)和乙酸钠(4.4g，53.9mmol，5.0eq)于EtOH(21mL)中的溶液中添加羟胺盐酸盐(3.7g，53.9mmol，5.0eq)。将反应混合物在90℃下搅拌6小时。在反应完成后，将反应溶液冷却至室温，添加3N盐酸以调整pH＝5，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。合并的有机层经Na2SO4干燥并浓缩，得到呈白色固体状的2-(7-溴苯并[d]异恶唑-3-基)乙酸(86-3)(2.1g，粗物质)。MS:m/z＝257[M+H]+。To a solution of 8-bromo-4-hydroxy-2H-benzopyran-2-one (86-2) (2.6 g, 10.7 mmol, 1.0 eq.) and sodium acetate (4.4 g, 53.9 mmol, 5.0 eq) in EtOH (21 mL) was added hydroxylamine hydrochloride (3.7 g, 53.9 mmol, 5.0 eq). The reaction mixture was stirred at 90 ° C for 6 hours. After the reaction was completed, the reaction solution was cooled to room temperature, 3N hydrochloric acid was added to adjust pH = 5, the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3). The combined organic layer was dried over Na 2 SO 4 and concentrated to give 2-(7-bromobenzo[d]isoxazol-3-yl)acetic acid (86-3) (2.1 g, crude material) as a white solid. MS: m/z=257[M+H] + .

向2-(7-溴苯并[d]异恶唑-3-基)乙酸(86-3)(2.1g，粗物质)于EtOH(20mL)中的溶液中添加5滴硫酸，将反应混合物在80℃下搅拌3小时。在反应完成后，浓缩，通过硅胶色谱(PE/EA＝4:1)纯化残余物，得到呈无色油状的2-(7-溴苯并[d]异恶唑-3-基)乙酸乙酯(86-4)(1.1g，两步骤产率：53.7％)。MS:m/z＝284[M+H]+。To a solution of 2-(7-bromobenzo[d]isoxazol-3-yl)acetic acid (86-3) (2.1 g, crude material) in EtOH (20 mL) was added 5 drops of sulfuric acid, and the reaction mixture was stirred at 80° C. for 3 hours. After the reaction was completed, it was concentrated and the residue was purified by silica gel chromatography (PE/EA=4:1) to give ethyl 2-(7-bromobenzo[d]isoxazol-3-yl)acetate (86-4) (1.1 g, two-step yield: 53.7%) as a colorless oil. MS: m/z=284[M+H] + .

向2-(7-溴苯并[d]异恶唑-3-基)乙酸乙酯(86-4)(1.1g，3.8mmol，1.0eq.)、双(频哪醇)二硼(1.27g，5.0mmol，1.3eq.)和KOAc(1.14g，11.6mmol，3.0eq.)于二恶烷(11mL)中的溶液中添加Pd(dppf)Cl2(288mg，0.38mmol，0.1eq.)，将反应混合物在100℃下在N2下搅拌16小时。在反应完成后，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取。分离有机层，经Na2SO4干燥并浓缩，得到呈黄色油状的2-(7-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊-2-基)苯并[d]异恶唑-3-基)乙酸乙酯(86-5)(1.01g，粗物质)。MS:m/z＝332[M+H]+。To a solution of ethyl 2-(7-bromobenzo[d]isoxazol-3-yl)acetate (86-4) (1.1 g, 3.8 mmol, 1.0 eq.), bis(pinacolato)diboron (1.27 g, 5.0 mmol, 1.3 eq.) and KOAc (1.14 g, 11.6 mmol, 3.0 eq.) in dioxane (11 mL) was added Pd(dppf) Cl2 (288 mg, 0.38 mmol, 0.1 eq.), and the reaction mixture was stirred at 100 °C under N2 for 16 h. After the reaction was completed, the mixture was diluted with H2O (50 mL) and extracted with EA (50 mL x 3). The organic layer was separated, dried over Na 2 SO 4 and concentrated to give ethyl 2-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]isoxazol-3-yl)acetate (86-5) (1.01 g, crude) as a yellow oil. MS: m/z=332 [M+H] + .

在0℃下，向2-(7-(4,4,5,5-四甲基-1,3,2-二氧杂硼杂环戊-2-基)苯并[d]异恶唑-3-基)乙酸乙酯(86-5)(1.01g，粗物质)于THF(10mL)和H2O(5mL)中的溶液中添加NaHCO3(63mg，7.6mmol，2.0eq.)和30％ H2O2(1.72g，15.2mmol，4.0eq.)，将反应混合物在室温下搅拌2小时。在反应完成后，用HCl水溶液(1N)将pH调整至5-6，将混合物用H2O(50mL)稀释并用EA(50mL×3)萃取，浓缩并通过逆相和硅胶柱(PE/EA＝4:1)纯化残余物，得到呈无色油状的2-(7-羟基苯并[d]异恶唑-3-基)乙酸乙酯(86-6)(702mg，两步骤产率：82％)。MS:m/z＝222[M+H]+。To a solution of ethyl 2-(7-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)benzo[d]isoxazol-3-yl)acetate (86-5) (1.01 g, crude) in THF (10 mL) and H2O (5 mL) at 0°C were added NaHCO3 ( 63 mg, 7.6 mmol, 2.0 eq.) and 30% H2O2 (1.72 g, 15.2 mmol, 4.0 eq.), and the reaction mixture was stirred at room temperature for 2 hours. After the reaction was completed, the pH was adjusted to 5-6 with aqueous HCl (1 N), the mixture was diluted with H 2 O (50 mL) and extracted with EA (50 mL×3), concentrated and the residue was purified by reverse phase and silica gel column (PE/EA=4:1) to give ethyl 2-(7-hydroxybenzo[d]isoxazol-3-yl)acetate (86-6) (702 mg, two-step yield: 82%) as a colorless oil. MS: m/z=222[M+H] + .

向2-(7-羟基苯并[d]异恶唑-3-基)乙酸乙酯(86-6)(702mg，3.17mmol，1.0eq.)和(R)-3-((甲苯磺酰基氧基)甲基)吡咯烷-1-甲酸叔丁酯(1.12g，3.17mmol，1.0eq.)于DMF(7mL)中的溶液中添加K2CO3(1.3g，9.51mmol，3.0eq.)。将反应混合物在80℃下搅拌16小时。在反应完成后，将混合物用H2O(20mL)稀释并用EA(20mL×3)萃取。将合并的有机层经Na2SO4干燥并浓缩。将残余物通过硅胶柱色谱(PE/EA＝4:1)纯化，得到呈白色固体状的(R)-3-(((3-(2-乙氧基-2-氧代乙基)苯并[d]异恶唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(86-7)(621mg，产率：48％)。MS:m/z＝405[M+H]+。To a solution of ethyl 2-(7-hydroxybenzo[d]isoxazol-3-yl)acetate (86-6) (702 mg, 3.17 mmol, 1.0 eq.) and tert-butyl (R)-3-((tosyloxy)methyl)pyrrolidine-1-carboxylate (1.12 g, 3.17 mmol, 1.0 eq.) in DMF (7 mL) was added K 2 CO 3 (1.3 g, 9.51 mmol, 3.0 eq.). The reaction mixture was stirred at 80° C. for 16 h. After the reaction was completed, the mixture was diluted with H 2 O (20 mL) and extracted with EA (20 mL×3). The combined organic layers were dried over Na 2 SO 4 and concentrated. The residue was purified by silica gel column chromatography (PE/EA=4:1) to give (R)-tert-butyl 3-(((3-(2-ethoxy-2-oxoethyl)benzo[d]isoxazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (86-7) (621 mg, yield: 48%) as a white solid. MS: m/z=405 [M+H] + .

在0℃下，向(R)-3-(((3-(2-乙氧基-2-氧代乙基)苯并[d]异恶唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(86-7)(621mg，1.53mmol，1.0eq.)于DMF(6mL)中的溶液中添加t-BuOK(171mg，1.53mmol，1.0eq.)和丙烯酰胺(108mg，1.53mmol，1.0eq.)，将反应混合物在室温下在N2下搅拌2小时。在反应完成后，将混合物用H2O(20mL)稀释并用EA(20mL×3)萃取。将 合并的有机层经Na2SO4干燥并浓缩。将残余物通过硅胶柱色谱(PE/EA＝1:2)纯化，得到呈白色固体状的(3R)-3-(((3-(2,6-二氧代哌啶-3-基)苯并[d]异恶唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(86-8)(95mg，产率：14.4％)。MS:m/z＝330[M+H-100]+。To a solution of (R)-tert-butyl 3-(((3-(2-ethoxy-2-oxoethyl)benzo[d]isoxazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (86-7) (621 mg, 1.53 mmol, 1.0 eq.) in DMF (6 mL) at 0°C were added t-BuOK (171 mg, 1.53 mmol, 1.0 eq.) and acrylamide (108 mg, 1.53 mmol, 1.0 eq.), and the reaction mixture was stirred at room temperature under N2 for 2 h. After the reaction was completed, the mixture was diluted with H2O (20 mL) and extracted with EA (20 mL×3). The combined organic layers were dried over Na2SO4 and concentrated. The residue was purified by silica gel column chromatography (PE/ EA =1:2) to give (3R)-tert-butyl 3-(((3-(2,6-dioxopiperidin-3-yl)benzo[d]isoxazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (86-8) (95 mg, yield: 14.4%) as a white solid. MS: m/z=330[M+H-100] + .

向(3R)-3-(((3-(2,6-二氧代哌啶-3-基)苯并[d]异恶唑-7-基)氧基)甲基)吡咯烷-1-甲酸叔丁酯(86-8)(95mg，0.22mmol，1.0eq.)于EA(2mL)中的溶液中添加4M HCl/EA(2mL)，将反应混合物在室温下搅拌3小时。在反应完成后，真空浓缩混合物，得到呈白色固体状的3-(7-(((R)-吡咯烷-3-基)甲氧基)苯并[d]异恶唑-3-基)哌啶-2,6-二酮盐酸盐(86-9)(71mg，粗物质)。粗物质未经纯化即用于下一步骤。MS:m/z＝330[M+H]+。To a solution of tert-butyl (3R)-3-(((3-(2,6-dioxopiperidin-3-yl)benzo[d]isoxazol-7-yl)oxy)methyl)pyrrolidine-1-carboxylate (86-8) (95 mg, 0.22 mmol, 1.0 eq.) in EA (2 mL) was added 4M HCl/EA (2 mL) and the reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was concentrated in vacuo to give 3-(7-(((R)-pyrrolidin-3-yl)methoxy)benzo[d]isoxazol-3-yl)piperidine-2,6-dione hydrochloride (86-9) (71 mg, crude) as a white solid. The crude material was used in the next step without purification. MS: m/z=330[M+H] + .

向3-(7-(((R)-吡咯烷-3-基)甲氧基)苯并[d]异恶唑-3-基)哌啶-2,6-二酮盐酸盐(86-9)(35mg，0.1mmol，1.0eq.)和N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-2-氟-4-(4-甲酰基哌啶-1-基)苯甲酰胺(53mg，0.1mmol，1.0eq.)于DMF(2mL)中的溶液中添加TEA(42mg，0.42mmol，4.0eq.)，将反应混合物在室温下搅拌2小时，然后向混合物中添加NaBH(OAc)3(32mg，0.15mmol，1.5eq.)并在室温下搅拌2小时。在反应完成后，将混合物通过反相色谱用MeCN:H2O＝56％洗脱纯化，得到呈白色固体状的N-((1r,4r)-4-((8-氰基喹啉-5-基)氧基)环己基)-4-(4-(((3R)-3-(((3-(2,6-二氧代哌啶-3-基)苯并[d]异恶唑-7-基)氧基)甲基)吡咯烷-1-基)甲基)哌啶-1-基)-2-氟苯甲酰胺(86)(20.1mg，产率：23.3％)。MS:m/z＝814[M+H]+。To a solution of 3-(7-(((R)-pyrrolidin-3-yl)methoxy)benzo[d]isoxazol-3-yl)piperidine-2,6-dione hydrochloride (86-9) (35 mg, 0.1 mmol, 1.0 eq.) and N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-2-fluoro-4-(4-formylpiperidin-1-yl)benzamide (53 mg, 0.1 mmol, 1.0 eq.) in DMF (2 mL) was added TEA (42 mg, 0.42 mmol, 4.0 eq.), the reaction mixture was stirred at room temperature for 2 hours, and then NaBH(OAc) 3 (32 mg, 0.15 mmol, 1.5 eq.) was added to the mixture and stirred at room temperature for 2 hours. After the reaction was completed, the mixture was purified by reverse phase chromatography eluting with MeCN:H 2 O=56% to give N-((1r,4r)-4-((8-cyanoquinolin-5-yl)oxy)cyclohexyl)-4-(4-(((3R)-3-(((3-(2,6-dioxopiperidin-3-yl)benzo[d]isoxazol-7-yl)oxy)methyl)pyrrolidin-1-yl)methyl)piperidin-1-yl)-2-fluorobenzamide (86) (20.1 mg, yield: 23.3%) as a white solid. MS: m/z=814[M+H] + .

1H NMR(400MHz,DMSO-d6)δ11.11(s,1H),9.08(d,J＝8.3Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8.29(d,J＝8.3Hz,1H),7.78–7.65(m,2H),7.51(t,J＝8.8Hz,1H),7.42(d,J＝7.9Hz,1H),7.33(dd,J＝8.3,6.3Hz,2H),7.26(d,J＝7.8Hz,1H),6.86–6.73(m,2H),4.75(brs,1H),4.61(dd,J＝12.0,4.9Hz,1H),4.39–4.21(m,2H),3.90(d,J＝13.0Hz,4H),3.71(brs,1H),3.25–3.12(m,3H),3.09–2.96(m,1H),2.91–2.73(m,4H),2.70–2.53(m,2H),2.31–2.12(m,4H),2.08–1.91(m,4H),1.87–1.80(m,2H),1.72–1.53(m,4H),1.32–1.20(m,2H). 1 H NMR (400MHz, DMSO-d 6) δ11.11(s,1H),9.08(d,J＝8.3Hz,1H),8.64(dd,J＝8.5,1.7Hz,1H),8.29(d,J＝8.3Hz,1H),7.78–7.65 (m,2H),7.51(t,J＝8.8H z,1H),7.42(d,J＝7.9Hz,1H),7.33(dd,J＝8.3,6.3Hz,2H),7.26(d,J＝7.8Hz,1H),6.86–6.73(m,2H ),4.75(brs,1H),4.61(dd, J＝12.0,4.9Hz,1H),4.39–4.21(m,2H),3.90(d,J＝13.0Hz,4H),3.71(brs,1H),3.25–3.12(m,3H),3.09–2.96 (m,1H),2.91–2. 73(m,4H),2.70–2.53(m,2H),2.31–2.12(m,4H),2.08–1.91(m,4H),1.87–1.80(m,2H),1.72–1.53(m,4H ),1.32–1.20(m,2H).

参照以上实施例的制备方法，制备以下化合物：







Referring to the preparation method of the above embodiment, the following compounds were prepared:







本发明化合物的核磁数据(H-NMR)如下：










The nuclear magnetic data (H-NMR) of the compounds of the present invention are as follows:










生物测试评价Biological test evaluation

以下结合测试例进一步描述解释本发明，但这些实施例并非意味着限制本发明的范围。The present invention is further described and explained below in conjunction with test examples, but these embodiments are not intended to limit the scope of the present invention.

测试例1 检测本发明化合物对人前列腺癌细胞LNCaP及VCaP中的AR蛋白降解水平Test Example 1 Detection of the AR protein degradation level of the compounds of the present invention in human prostate cancer cells LNCaP and VCaP

细胞培养，人前列腺癌细胞LNCaP及VCaP，在培养基(90％1640培养基,10％ FBS胎牛血清,1×P/S抗生素)中传代培养。Cell culture, human prostate cancer cells LNCaP and VCaP were subcultured in culture medium (90% 1640 medium, 10% FBS fetal bovine serum, 1× P/S antibiotics).

细胞接种和化合物处理：Cell seeding and compound treatment:

1.细胞培养过程中保证活力>90％,将培养的细胞消化后，在培养基中重悬稀释至100000个细胞每毫升，将100μL细胞种入细胞培养板中10000个细胞每孔，在细胞培养箱37℃，5％CO2下孵育4-8小时。1. Ensure that the cell viability is >90% during cell culture. Digest the cultured cells, resuspend and dilute them to 100,000 cells per ml in the culture medium, seed 100 μL of cells into 10,000 cells per well of the cell culture plate, and incubate them in a cell culture incubator at 37°C and 5% CO2 for 4-8 hours.

2.用DMSO4倍梯度稀释测试化合物(300μM到18nM)。将1μL测试化合物加入细胞培养板中(化合物终浓度3000nM到0.18nM)，阳性参照孔为3000nM ARV110，阴性参照孔为无化合物的DMSO。2. Dilute the test compound 4-fold with DMSO (300μM to 18nM). Add 1μL of the test compound to the cell culture plate (final compound concentration 3000nM to 0.18nM), the positive reference well is 3000nM ARV110, and the negative reference well is DMSO without compound.

3.在培养箱中37℃，5％ CO2下孵育8小时。3. Incubate in an incubator at 37°C, 5% CO2 for 8 hours.

Elisa检测：Elisa test:

1.弃去培养基，添加1X细胞裂解缓冲液处理细胞。将100uL上清液转移到包被有捕获抗体(Elisa检测用各试剂购自Cell Signaling Technology，货号12850)的微孔板中。1. Discard the culture medium and add 1X cell lysis buffer to treat the cells. Transfer 100uL of the supernatant to a microplate coated with capture antibody (ELISA test reagents purchased from Cell Signaling Technology, Cat. No. 12850).

2. 4℃下放置过夜，弃去孔中液体，用1X洗涤缓冲液洗涤4次，每次200μL。向每个孔中加入100μL重组检测抗体，将板在37℃孵育1小时。2. Place at 4°C overnight, discard the liquid in the wells, wash 4 times with 1X wash buffer, 200 μL each time. Add 100 μL of recombinant detection antibody to each well and incubate the plate at 37°C for 1 hour.

3.重复洗涤程序。向每个孔中加入100μL重组的HRP连接的二抗，将板在37℃孵育30分钟。3. Repeat the washing procedure. Add 100 μL of recombinant HRP-linked secondary antibody to each well and incubate the plate at 37°C for 30 minutes.

4.重复洗涤程序。向每个孔中加入100μL TMB底物。4. Repeat the washing procedure. Add 100 μL of TMB substrate to each well.

5.孵育5-15分钟后，Envision仪器上读取光吸收值OD450值。5. After incubation for 5-15 minutes, read the absorbance OD450 value on the Envision instrument.

6.利用OD450值计算降解率。
％Degradation＝100-(化合物孔OD值-阳性参照孔OD值)/(阴性参照孔OD值-阳性参照孔OD
值)×100.6. Calculate the degradation rate using the OD450 value.
%Degradation = 100 - (OD value of compound well - OD value of positive reference well) / (OD value of negative reference well - OD value of positive reference well)
value)×100.

用Graphpad 8.0软件4参数模型计算DC50值。结果如表1、表1-1和表2所示： The DC 50 values were calculated using the Graphpad 8.0 software 4-parameter model. The results are shown in Table 1, Table 1-1 and Table 2:

表1.本发明化合物100nM、10nM浓度下LNCaP细胞AR蛋白降解率
Table 1. AR protein degradation rate of LNCaP cells at 100 nM and 10 nM concentrations of the compounds of the present invention

表1-1.本发明化合物30nM、3nM浓度下VCaP细胞AR蛋白降解率
Table 1-1. AR protein degradation rate of VCaP cells at 30 nM and 3 nM concentrations of the compounds of the present invention

表2.本发明化合物对VCaP细胞AR蛋白降解率
Table 2. AR protein degradation rate of the compounds of the present invention on VCaP cells

测试例2人前列腺癌细胞VCaP，Western blotting测定化合物诱导的AR降解Test Example 2 Human prostate cancer cell VCaP, Western blotting to determine compound-induced AR degradation

1.细胞培养1. Cell Culture

复苏细胞于T25瓶中，DMEM Medium+10％FBS+1％P/S培养，当细胞汇合度达到80-90％，进行传代。试验前，将VCaP细胞接种到96孔细胞培养板中，每个孔接种3×105个细胞，加入100μL培养基。将细胞置于37℃，5％ CO2培养箱中培养过夜。The revived cells were cultured in T25 flasks with DMEM Medium + 10% FBS + 1% P/S. When the cell confluence reached 80-90%, the cells were passaged. Before the experiment, VCaP cells were inoculated into 96-well cell culture plates, 3×10 5 cells were inoculated into each well, and 100 μL of culture medium was added. The cells were cultured in a 37°C, 5% CO 2 incubator overnight.

2.药物处理2. Drug treatment

使用IDOT仪器加化合物至96孔板中，每孔加入稀释好的化合物100nL。细胞培养皿在37℃，5％CO2培养箱中培养8小时。Add the compound to the 96-well plate using an IDOT instrument, adding 100 nL of the diluted compound to each well. The cell culture dish was cultured in a 37°C, 5% CO 2 incubator for 8 hours.

3.试剂配置3. Reagent Configuration

a)细胞裂解液：向20mL细胞裂解液中，加入一片蛋白酶抑制剂，以及一片磷酸酶抑制剂，轻轻混匀至完全溶解。a) Cell lysis buffer: Add one tablet of protease inhibitor and one tablet of phosphatase inhibitor to 20 mL of cell lysis buffer and mix gently until completely dissolved.

b)4×制样工作液：向4×制样缓冲液中加入终浓度200mM DTT，配置成4×制样工作液。b) 4× sample preparation working solution: Add DTT to a final concentration of 200 mM into 4× sample preparation buffer to prepare 4× sample preparation working solution.

c)电泳缓冲液：用超纯水稀释20×电泳缓冲液至1×。 c) Electrophoresis buffer: Dilute 20× electrophoresis buffer to 1× with ultrapure water.

d)转膜缓冲液：用超纯水稀释10×转膜缓冲液至1×，并加入20％甲醇。d) Transfer buffer: Dilute 10× transfer buffer to 1× with ultrapure water and add 20% methanol.

e)TBST缓冲液：用超纯水稀释10×电泳缓冲液至1×。e) TBST buffer: dilute 10× electrophoresis buffer to 1× with ultrapure water.

f)封闭液：5g BSA用1×TBST溶解至100mL，4℃保存。f) Blocking solution: Dissolve 5 g BSA in 1×TBST to 100 mL and store at 4°C.

g)一抗工作液：g) Primary antibody working solution:

AR一抗工作液：在24mL封闭液中加入Androgen Receptor(D6F11)Rabbit mAb#5153抗体24μL；AR primary antibody working solution: add Androgen Receptor (D6F11) to 24 mL of blocking solution Rabbit mAb #5153 antibody 24 μL;

β-actin一抗工作液：在24mL封闭液中加入β-Actin(13E5)Rabbit mAb#4970抗体2.4μL；β-actin primary antibody working solution: add 2.4 μL of β-Actin (13E5) Rabbit mAb #4970 antibody to 24 mL of blocking solution;

h)二抗工作液：将Anti-rabbit IgG，HRP-linked antibody用封闭液分别稀释5000倍和10000倍。h) Secondary antibody working solution: dilute Anti-rabbit IgG and HRP-linked antibody 5000 times and 10000 times respectively with blocking solution.

4.蛋白提取4. Protein Extraction

吸尽上清液后，倒置离心300rpm，30s，每孔加入细胞裂解液45μL，冰上静置20分钟。吸取细胞裂解液至新离心管中，每个样本加入15μL 4×制样工作液，70℃加热10分钟。剩余蛋白样品置于-80℃保存。After the supernatant was completely aspirated, the tube was centrifuged inverted at 300 rpm for 30 seconds. 45 μL of cell lysis solution was added to each well and the tube was placed on ice for 20 minutes. The cell lysis solution was aspirated into a new centrifuge tube, 15 μL of 4× sample preparation working solution was added to each sample, and the tube was heated at 70°C for 10 minutes. The remaining protein sample was stored at -80°C.

5.Western Blot试验5. Western Blot test

取蛋白预制胶，每孔上样10μL，恒压120V进行电泳，60分钟。使用PVDF膜进行转膜，恒流300mA，80分钟。转膜后，用封闭液室温封闭60分钟。一抗工作液4℃孵育过夜。用1×TBST缓冲液洗膜，洗三次，每次10分钟。二抗工作液室温孵育1小时。用1×TBST缓冲液洗膜，洗三次，每次10分钟。用显色液进行曝光显色。数据处理和分析。扫膜仪扫描条带后，用image J分析目的基因AR条带和内参基因β-actin条带的灰度，AR/β-actin的比值作为校正后的样品孔AR的含量，以不加化合物的样品孔AR的含量作为100％ AR量，即0％ AR Degradation。其他样品孔除以不加化合物的样品孔AR的含量，来计算降解率。用Graphpad 8.0软件4参数模型计算DC50值。结果如表3所示。Take the protein precast gel, load 10μL per well, and perform electrophoresis at a constant voltage of 120V for 60 minutes. Use PVDF membrane for transfer, constant current 300mA, 80 minutes. After transfer, block with blocking solution at room temperature for 60 minutes. Incubate with primary antibody working solution at 4℃ overnight. Wash the membrane with 1×TBST buffer three times, 10 minutes each time. Incubate with secondary antibody working solution at room temperature for 1 hour. Wash the membrane with 1×TBST buffer three times, 10 minutes each time. Expose and color with color developing solution. Data processing and analysis. After scanning the bands with a film scanner, use image J to analyze the grayscale of the target gene AR band and the internal reference gene β-actin band. The ratio of AR/β-actin is used as the corrected AR content of the sample well, and the AR content of the sample well without compound is used as 100% AR content, that is, 0% AR Degradation. The degradation rate is calculated by dividing the AR content of the sample well without compound by the AR content of the sample well. The DC50 values were calculated using a 4-parameter model using Graphpad 8.0 software. The results are shown in Table 3.

表3.本发明化合物对VCaP细胞AR蛋白降解率
Table 3. AR protein degradation rate of the compounds of the present invention on VCaP cells

测试例3小鼠药代动力学试验Test Example 3: Pharmacokinetics test in mice

采用雄性ICR小鼠，禁食过夜。取6只小鼠，给药前称重，根据体重计算给药量，通过静脉注射或灌胃口服给药各3只小鼠。化合物溶媒为5％DMSO+10％solutol+85％saline。化合物口服灌胃给药3mg/kg，尾静脉注射给药1mg/kg。给药后IV:0.083h，0.25h，0.5h，1h，2h，4h，8h，24h。PO:0.25h， 0.5h，1h，2h，4h，6h,8h，24h。经颈静脉采血，每个样品采集约0.03mL，肝素钠抗凝，采集后放置冰上。血液样本采集后置于冰上，并于1小时之内离心分离血浆(离心条件：6800g，6分钟，2-8℃)。血浆样本在分析前存放时则放于-80℃冰箱内。分析样品的同时进行质控样品的日内准确度评价，并要求超过66.7％的质控样品的准确度在80-120％之间。通过不同时间点的血药浓度数据，运用Phoenix WinNonlin7.0计算药代动力学参数，提供AUC0-t、AUC0-∞、MRT0-∞、Cmax、Tmax、和T1/2等参数及其平均值和标准差。实验结果如表4。Male ICR mice were fasted overnight. Six mice were weighed before administration, and the dosage was calculated based on the body weight. Three mice were administered intravenously or orally. The compound solvent was 5% DMSO + 10% solutol + 85% saline. The compound was administered orally by gavage at 3 mg/kg and by tail vein injection at 1 mg/kg. IV after administration: 0.083h, 0.25h, 0.5h, 1h, 2h, 4h, 8h, 24h. PO: 0.25h, 0.5h, 1h, 2h, 4h, 6h, 8h, 24h. Blood was collected from the jugular vein, and about 0.03mL of each sample was collected. Heparin sodium was used for anticoagulation and placed on ice after collection. After blood samples were collected, they were placed on ice and centrifuged within 1 hour to separate plasma (centrifugation conditions: 6800g, 6 minutes, 2-8℃). Plasma samples were stored in a -80℃ refrigerator before analysis. While analyzing the samples, the intra-day accuracy of the quality control samples was evaluated, and the accuracy of more than 66.7% of the quality control samples was required to be between 80-120%. Phoenix WinNonlin7.0 was used to calculate the pharmacokinetic parameters based on the blood drug concentration data at different time points, providing parameters such as AUC0-t, AUC0-∞, MRT0-∞, Cmax, Tmax, and T1/2 and their mean and standard deviation. The experimental results are shown in Table 4.

表4.本发明化合物的PK数据
Table 4. PK data of the compounds of the present invention

实验结果表明，本发明化合物具有良好的生物利用度，具有优良的药代动力学性质。 The experimental results show that the compound of the present invention has good bioavailability and excellent pharmacokinetic properties.

Claims (13)

一种通式(I)所示化合物、其立体异构体或其药学上可接受的盐：
A compound represented by general formula (I), its stereoisomer or a pharmaceutically acceptable salt thereof:
其中：in: M1选自N或CR1；M 1 is selected from N or CR 1 ; M2选自N或CR2；M 2 is selected from N or CR 2 ; M3选自N或CR3；M 3 is selected from N or CR 3 ; M4选自N或CR4；M 4 is selected from N or CR 4 ; M5选自N或CR5；M 5 is selected from N or CR 5 ; R1、R2、R3、R4和R5各自独立的选自氢、氘、卤素、氰基、烷基、烷氧基、卤代烷基、卤代烷氧基、羟基烷基、羟基烷氧基、环烷基或杂环基；R 1 , R 2 , R 3 , R 4 and R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, alkyl, alkoxy, haloalkyl, haloalkoxy, hydroxyalkyl, hydroxyalkoxy, cycloalkyl or heterocyclyl; 环A、环B和环C各自独立的选自环烷基、杂环基、芳基或杂芳基；Ring A, Ring B and Ring C are each independently selected from cycloalkyl, heterocyclyl, aryl or heteroaryl; L1选自键、-C(O)NH-、-NH-、-O-、-NHC(O)-、-C(O)-、-S(＝O)-、-S(＝O)2-、-CH2-或-(CH2)2-；L 1 is selected from a bond, -C(O)NH-, -NH-, -O-, -NHC(O)-, -C(O)-, -S(=O)-, -S(=O) 2 -, -CH 2 - or -(CH 2 ) 2 -; L2为-Ak1-Cy1-Ak2-Cy2-Ak3-， L2 is -Ak1-Cy1-Ak2-Cy2-Ak3-, Ak1、Ak2和Ak3各自独立的选自键、-(CH2)n1-O-(CH2)n2-、-(CH2)n3-、-(CH2)n4-S-(CH2)n5、-(CH2)n6-C(O)NH-(CH2)n7-、-(CH2)n8-NH-(CH2)n9、-(CH2)n10-NHC(O)-(CH2)n11、-(CH2)n12-C(O)-(CH2)n13、-(CH2)n14-S(＝O)-(CH2)n15-或-(CH2)n16-S(＝O)2-(CH2)n17-，所述的-(CH2)n1-O-(CH2)n2-、-(CH2)n3-、-(CH2)n4-S-(CH2)n5、-(CH2)n6-C(O)NH-(CH2)n7-、-(CH2)n8-NH-(CH2)n9、-(CH2)n10-NHC(O)-(CH2)n11、-(CH2)n12-C(O)-(CH2)n13、-(CH2)n14-S(＝O)-(CH2)n15-和-(CH2)n16-S(＝O)2-(CH2)n17-，任选的可以进一步被取代；Ak1, Ak2 and Ak3 are each independently selected from a bond, -(CH 2 ) n1 -O-(CH 2 ) n2 -, -(CH 2 ) n3 -, -(CH 2 ) n4 -S-(CH 2 ) n5 , -(CH 2 ) n6 -C(O)NH-(CH 2 ) n7 -, -(CH 2 ) n8 -NH-(CH 2 ) n9 , -(CH 2 ) n10 -NHC(O)-(CH 2 ) n11 , -(CH 2 ) n12 -C(O)-(CH 2 ) n13 , -(CH 2 ) n14 -S(＝O)-(CH 2 ) n15 - or -(CH 2 ) n16 -S(＝O) 2 -(CH 2 ) n17 -, wherein -(CH 2 ) n1 -O-(CH 2 ) n2 - 2 ) n2 -, -(CH 2 ) n3 -, -(CH 2 ) n4 -S-(CH 2 ) n5 , -(CH 2 ) n6 -C(O)NH-(CH 2 ) n7 -, -(CH 2 ) n8 -NH-(CH 2 ) n9 , -(CH 2 ) n10 -NHC(O)-(CH 2 ) n11 , -(CH 2 ) n12 -C(O)-(CH 2 ) n13 , -(CH 2 ) n14 -S(═O)-(CH 2 ) n15 - and -(CH 2 ) n16 -S(═O) 2 -(CH 2 ) n17 -, which may be further substituted; Cy1和Cy2各自独立的选自键、亚环己基、亚哌啶基、亚哌嗪基、亚氮杂环丁烷基、亚氮杂环戊烷基、亚环丁烷基或亚环戊烷基，所述亚环己基、亚哌啶基、亚哌嗪基、亚氮杂环丁烷基、亚氮杂环戊烷基、亚环丁烷基和亚环戊烷基，任选的可以进一步被取代；或者，所述亚环己基、亚哌啶基、亚哌嗪基、亚氮杂环丁烷基、亚氮杂环戊烷基、亚环丁烷基和亚环戊烷基上的任意两个原子可以链接形成环烷基或杂环基；Cy1 and Cy2 are each independently selected from a bond, a cyclohexylene, a piperidinylene, a piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene or a cyclopentylene, and the cyclohexylene, piperidinylene, piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene and a cyclopentylene may optionally be further substituted; or, any two atoms on the cyclohexylene, piperidinylene, piperazinylene, an azetidinylene, an azocyclopentylene, a cyclobutylene and a cyclopentylene may be linked to form a cycloalkyl or heterocyclic group; Ra、Rb和Rc各自独立的选自氢、氘、卤素、氧代基、氰基、羟基、烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基、杂环基、芳基或杂芳基，所述烷基、烷氧基、卤代烷基、卤代烷氧基、环烷基、杂环基、芳基和杂芳基，任选的可以进一步被取代； Ra , Rb and Rc are each independently selected from hydrogen, deuterium, halogen, oxo, cyano, hydroxy, alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl or heteroaryl, and the alkyl, alkoxy, haloalkyl, haloalkoxy, cycloalkyl, heterocyclyl, aryl and heteroaryl may be further substituted; 或者，任意两个Rc链接形成环烷基、杂环基、芳基或杂芳基，所述环烷基、杂环基、芳基和杂芳基，任选的可以进一步被取代； Alternatively, any two R c are linked to form a cycloalkyl group, a heterocyclic group, an aryl group or a heteroaryl group, and the cycloalkyl group, the heterocyclic group, the aryl group and the heteroaryl group may be further substituted; 或者，Ra与L1链接形成环烷基、杂环基或杂芳基，所述环烷基、杂环基和杂芳基，任选的可以进一步被取代；Alternatively, R a and L 1 are linked to form a cycloalkyl group, a heterocyclic group or a heteroaryl group, and the cycloalkyl group, the heterocyclic group and the heteroaryl group may be further substituted; x、y和z各自独立的选自0、1、2或3；x, y and z are each independently selected from 0, 1, 2 or 3; n1、n2、n3、n4、n5、n6、n7、n8、n9、n10、n11、n12、n13、n14、n15、n16和n17各自独立的选自0、1、2或3。n1, n2, n3, n4, n5, n6, n7, n8, n9, n10, n11, n12, n13, n14, n15, n16 and n17 are each independently selected from 0, 1, 2 or 3. 根据权利要求1所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述通式(I)进一步如通式(II)所示：
The compound according to claim 1, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the general formula (I) is further represented by the general formula (II):
其中：Rd选自氢、卤素或C1-3烷基；p1和p2各自独立的选自0、1、2、3或4；L4选自键、-CH2-、-NH-、-O-、-S-或-C(O)-。wherein: R d is selected from hydrogen, halogen or C 1-3 alkyl; p1 and p2 are each independently selected from 0, 1, 2, 3 or 4; L 4 is selected from a bond, -CH 2 -, -NH-, -O-, -S- or -C(O)-. 根据权利要求1或2所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述环A选自苯基、5-6元单环杂芳基或9-10元双环杂环基；The compound according to claim 1 or 2, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the ring A is selected from phenyl, 5-6 membered monocyclic heteroaryl or 9-10 membered bicyclic heterocyclic group; 优选的，环A选自苯基、吡唑基、吡咯基、吡啶基、嘧啶基、苯并哒嗪酮基、苯并吡唑基或苯并吡咯烷基；Preferably, ring A is selected from phenyl, pyrazolyl, pyrrolyl, pyridyl, pyrimidinyl, benzopyridazinone, benzopyrazolyl or benzopyrrolidinyl; 或者，环B选自环己烷基或环丁烷基；Alternatively, Ring B is selected from cyclohexyl or cyclobutane; 或者，环C选自苯基、5-6元单环杂芳基、9-10元双环杂环基、9-10元双环杂芳基；Alternatively, ring C is selected from phenyl, 5-6 membered monocyclic heteroaryl, 9-10 membered bicyclic heterocyclyl, 9-10 membered bicyclic heteroaryl; 优选的，环C选自苯并吡啶基、苯基、吡啶基、苯并噻唑基、苯并吡嗪基、吡啶并吡唑基、吡啶并三唑基、吡啶并苯基、苯并噻二唑或苯并哒嗪基。Preferably, ring C is selected from benzopyridyl, phenyl, pyridyl, benzothiazolyl, benzopyrazinyl, pyridopyrazolyl, pyridotriazolyl, pyridophenyl, benzothiadiazole or benzopyridazinyl. 根据权利要求1或3所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述化合物进一步如通式(II-A)所示：
The compound according to claim 1 or 3, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the compound is further represented by general formula (II-A):
其中：in: 选自 且， Selected from and, 当选自时，环A不为 when Selected from When ring A is not 根据权利要求1-4任一项所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，环A选自苯基、吡啶基、 The compound according to any one of claims 1 to 4, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that ring A is selected from phenyl, pyridyl, 根据权利要求1-4任一项所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述Ra、Rb和Rc各自独立的选自氢、氘、卤素、氰基、羟基、氧代基、C1-3烷基、C1-3烷氧基、羟基C1-3烷基、羟基C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C3-4环烷基或3-4元杂环基。The compound according to any one of claims 1 to 4, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the Ra , Rb and Rc are each independently selected from hydrogen, deuterium, halogen, cyano, hydroxyl, oxo, C1-3 alkyl, C1-3 alkoxy, hydroxyC1-3 alkyl, hydroxyC1-3 alkoxy, C1-3 haloalkyl, C1-3 haloalkoxy, C3-4 cycloalkyl or 3-4 membered heterocyclic group. 根据权利要求1-5任一项所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述R1、R2、R3、R4和R5各自独立的选自氢、氘、卤素、氰基、C1-3烷基、C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、羟基C1-3烷基、羟基C1-3烷氧基、C3-4环烷基或3-4元杂环基。The compound according to any one of claims 1 to 5, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that R 1 , R 2 , R 3 , R 4 and R 5 are each independently selected from hydrogen, deuterium, halogen, cyano, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, hydroxy C 1-3 alkyl, hydroxy C 1-3 alkoxy, C 3-4 cycloalkyl or 3-4 membered heterocyclic group. 根据权利要求1-6任一项所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述L1选自键、-C(O)NH-、-NH-、-O-、-NHC(O)-、-C(O)-、-CH2-或-(CH2)2-；The compound, stereoisomer or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6, characterized in that L 1 is selected from a bond, -C(O)NH-, -NH-, -O-, -NHC(O)-, -C(O)-, -CH 2 - or -(CH 2 ) 2 -; L2选自 L 2 is selected from 根据权利要求1或2所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述化合物进一步如通式(II-B)所示：
The compound according to claim 1 or 2, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the compound is further represented by general formula (II-B):
其中：in: L3和L4各自独立的选自键、-CH2-、-NH-、-O-、-S-或-C(O)-；L 3 and L 4 are each independently selected from a bond, -CH 2 -, -NH-, -O-, -S- or -C(O)-; M6和M7各自独立的选自-CR6R7-或-C(O)-；M 6 and M 7 are each independently selected from -CR 6 R 7 - or -C(O)-; M8选自N或CR8；M 8 is selected from N or CR 8 ; R6、R7和R8各自独立的选自氢、氘、氟、氯、羟基、氰基、C1-3烷基、羟基C1-3烷基、C1-3烷氧基、羟基C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C3-4环烷基或3-4元杂环基；R 6 , R 7 and R 8 are each independently selected from hydrogen, deuterium, fluorine, chlorine, hydroxyl, cyano, C 1-3 alkyl, hydroxy C 1-3 alkyl, C 1-3 alkoxy, hydroxy C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, C 3-4 cycloalkyl or 3-4 membered heterocyclyl; Rd选自氢、卤素或C1-3烷基；R d is selected from hydrogen, halogen or C 1-3 alkyl; j、p1和p2各自独立的选自0、1、2、3或4。j, p1 and p2 are each independently selected from 0, 1, 2, 3 or 4. 根据权利要求2、3、5-7任一项所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述通式(II)化合物进一步如通式(II-C)所示：
The compound according to any one of claims 2, 3, 5-7, its stereoisomer or pharmaceutically acceptable salt thereof, characterized in that the compound of general formula (II) is further represented by general formula (II-C):
其中：M2选自N或CR2；M3选自N或CR3；M4选自N或CR4；M5选自N或CR5；环B选自环已烷基或环丁烷基；环C选自苯基、5-6元单环杂芳基或10元双环杂芳基；优选地，环C选自苯基、吡啶基或苯并吡啶基；Ra选自氢、卤素或C1-3烷基；Rd选自氢、卤素或C1-3烷基；Rb各自独立地选自氢、C1-3烷基、C1-3烷氧基或C1-3羟基烷基；Rc各自独立地选自氢、氰基、卤素、羟基、C1-3烷基、C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C1-3羟基烷基或C1-3羟基烷氧基；R2、R3、R4和R5 各自独立地选自氢、卤素、氰基、C1-3烷基、C1-3烷氧基、C1-3卤代烷基、C1-3卤代烷氧基、C1-3羟基烷基或C1-3羟基烷氧基；y和z各自独立的选自0、1、2或3。wherein: M2 is selected from N or CR2 ; M3 is selected from N or CR3 ; M4 is selected from N or CR4 ; M5 is selected from N or CR5 ; Ring B is selected from cyclohexyl or cyclobutane; Ring C is selected from phenyl, 5-6-membered monocyclic heteroaryl or 10-membered bicyclic heteroaryl; preferably, Ring C is selected from phenyl, pyridyl or benzopyridyl; Ra is selected from hydrogen, halogen or C1-3 alkyl; Rd is selected from hydrogen, halogen or C1-3 alkyl; Rb is each independently selected from hydrogen, C1-3 alkyl, C1-3 alkoxy or C1-3 hydroxyalkyl; Rc is each independently selected from hydrogen, cyano, halogen, hydroxyl, C1-3 alkyl, C1-3 alkoxy, C1-3 haloalkyl, C1-3 haloalkoxy, C1-3 hydroxyalkyl or C1-3 hydroxyalkoxy; R2 , R3 , R4 and R5 are Each is independently selected from hydrogen, halogen, cyano, C 1-3 alkyl, C 1-3 alkoxy, C 1-3 haloalkyl, C 1-3 haloalkoxy, C 1-3 hydroxyalkyl or C 1-3 hydroxyalkoxy; y and z are each independently selected from 0, 1, 2 or 3. 根据权利要求1-10任一项所述的化合物、其立体异构体或其药学上可接受的盐，其特征在于，所述化合物选自表5化合物。The compound according to any one of claims 1 to 10, its stereoisomer or a pharmaceutically acceptable salt thereof, characterized in that the compound is selected from the compounds in Table 5. 一种药物组合物，包括权利要求1-11任一项所述的化合物、其立体异构体或其药学上可接受的盐，以及药学上可接受的载体。A pharmaceutical composition comprising the compound according to any one of claims 1 to 11, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. 根据权利要求1-11任一项所述的化合物、其立体异构体或其药学上可接受的盐，以及权利要求12所述的组合物，在制备雄激素受体调控的相关疾病的药物中的应用。 Use of the compound according to any one of claims 1 to 11, its stereoisomer or a pharmaceutically acceptable salt thereof, and the composition according to claim 12 in the preparation of a drug for androgen receptor-regulated related diseases.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12496301B2 (en) 2024-12-06 2025-12-16 Arvinas Operations, Inc. Use of androgen receptor degrader for the treatment of spinal and bulbar muscular atrophy

Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104684896A (en) * 2012-06-26 2015-06-03 拜耳医药股份有限公司 N-[4-(quinolin-4-yloxy)cyclohexyl(methyl)](hetero)arylcarboxamides as androgen receptor antagonists, production and use thereof as medicinal products
CN110506039A (en) * 2016-10-11 2019-11-26 阿尔维纳斯股份有限公司 Compounds and methods for targeted degradation of androgen receptor
US20210087171A1 (en) * 2019-09-23 2021-03-25 Accutar Biotechnology Inc. Novel substituted quinoline-8-carbonitrile derivatives with androgen receptor degradation activity and uses thereof
CN113329999A (en) * 2019-09-23 2021-08-31 冰洲石生物科技公司 Novel substituted quinoline-8-carbonitrile derivatives having androgen receptor degrading activity and uses thereof
CN114466850A (en) * 2019-08-12 2022-05-10 拜耳股份有限公司 [1,2,4] triazolo [1,5-C ] quinazolin-5-amines
CN115974840A (en) * 2017-01-31 2023-04-18 阿尔维纳斯运营股份有限公司 Human cerebellar protein ligands and bifunctional compounds containing them
WO2023180388A1 (en) * 2022-03-24 2023-09-28 Glaxosmithkline Intellectual Property Development Limited 2,4-dioxotetrahydropyrimidinyl derivatives as degrons in protacs
WO2024102784A1 (en) * 2022-11-09 2024-05-16 Accutar Biotechnology, Inc. Substituted quinolinone-8-carbonitrile derivatives having androgen degradation activity and uses thereof

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104684896A (en) * 2012-06-26 2015-06-03 拜耳医药股份有限公司 N-[4-(quinolin-4-yloxy)cyclohexyl(methyl)](hetero)arylcarboxamides as androgen receptor antagonists, production and use thereof as medicinal products
CN110506039A (en) * 2016-10-11 2019-11-26 阿尔维纳斯股份有限公司 Compounds and methods for targeted degradation of androgen receptor
CN115974840A (en) * 2017-01-31 2023-04-18 阿尔维纳斯运营股份有限公司 Human cerebellar protein ligands and bifunctional compounds containing them
CN114466850A (en) * 2019-08-12 2022-05-10 拜耳股份有限公司 [1,2,4] triazolo [1,5-C ] quinazolin-5-amines
US20210087171A1 (en) * 2019-09-23 2021-03-25 Accutar Biotechnology Inc. Novel substituted quinoline-8-carbonitrile derivatives with androgen receptor degradation activity and uses thereof
CN113329999A (en) * 2019-09-23 2021-08-31 冰洲石生物科技公司 Novel substituted quinoline-8-carbonitrile derivatives having androgen receptor degrading activity and uses thereof
WO2023180388A1 (en) * 2022-03-24 2023-09-28 Glaxosmithkline Intellectual Property Development Limited 2,4-dioxotetrahydropyrimidinyl derivatives as degrons in protacs
WO2024102784A1 (en) * 2022-11-09 2024-05-16 Accutar Biotechnology, Inc. Substituted quinolinone-8-carbonitrile derivatives having androgen degradation activity and uses thereof

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12496301B2 (en) 2024-12-06 2025-12-16 Arvinas Operations, Inc. Use of androgen receptor degrader for the treatment of spinal and bulbar muscular atrophy

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