WO2024051755A1 - Kif18a抑制剂化合物、药物组合物及其制备方法和应用 - Google Patents

Kif18a抑制剂化合物、药物组合物及其制备方法和应用 Download PDF

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WO2024051755A1
WO2024051755A1 PCT/CN2023/117322 CN2023117322W WO2024051755A1 WO 2024051755 A1 WO2024051755 A1 WO 2024051755A1 CN 2023117322 W CN2023117322 W CN 2023117322W WO 2024051755 A1 WO2024051755 A1 WO 2024051755A1
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ring
compound
alkyl
cyano
alkoxy
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PCT/CN2023/117322
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English (en)
French (fr)
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陆标
杨方龙
张辰
王思勤
金磊
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长春金赛药业有限责任公司
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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
    • A61K31/438—The ring being spiro-condensed with carbocyclic or heterocyclic ring systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4427—Non condensed pyridines; Hydrogenated derivatives thereof containing further heterocyclic ring systems
    • 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
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • 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 requires the following:
  • the invention belongs to the field of medicine, and specifically relates to a KIF18A inhibitor compound, a pharmaceutical composition and a preparation method and application thereof.
  • Cancer is one of the most serious diseases affecting human health, with mortality and morbidity often ranking among the top among various diseases. Although the quality of life of some patients has been greatly improved with the continuous development and progress of medical technology and drug research and development, there are still more unidentified drugs on the road to finding effective treatments or cures for different cancers. meet clinical needs, and more new targets will provide new possibilities for future cancer drug development.
  • Unregulated cell proliferation in cancer cells occurs due to damage or deletion of one or more genes that regulate the cell cycle.
  • Various kinases and kinesins have been identified to play critical roles in cell cycle and mitotic regulation and progression in normally dividing cells and cancer cells.
  • Kinesin molecules are dynein molecules that use intracellular microtubules as tracks. They are also called molecular motors. They can convert ATP energy into mechanical energy. They are used in eukaryotic cells and cell mitosis and meiosis, the growth and development of tissues and organs, and the growth of neurons. Development and signal transduction are closely related. Kinesin members share a relatively conserved motor domain.
  • N-type kinesin that is, the amino (-NH 2 ) terminal region of the protein polypeptide chain has a motor domain
  • M-type kinesin that is, the middle region Has a motor domain
  • C-type kinesin that is, the carboxyl (-COOH) terminal region has a motor domain.
  • Chromosomal instability is a hallmark of cancer and results from errors in chromosome segregation during mitosis.
  • Targeting chromosomal instability is an emerging therapeutic strategy in drug development.
  • KIF18A a member of the N-type Kinesin-8 kinesin family, has been shown to play a role in maintaining bipolar spindle integrity and promoting the viability of cancer cells with chromosomal instability.
  • Mitosis is an effective point of intervention, and many antimitotic drugs are used clinically to treat human cancers. The most widely used tubulin inhibitor, it can both stabilize microtubules and prevent microtubule assembly. Current anti-mitotic drugs have limitations such as narrow therapeutic windows, and these problems require the development of new targets to solve.
  • tubulin inhibitors are widely used as standard treatments for many human cancer types, these drugs are associated with collateral damage to normal cells, including myelosuppression and neurotoxicity. Since KIF18A may not be essential in normal diploid somatic cells (KIF18A knockout mice are viable but defective in reproduction, suggesting that KIF18A is not an essential gene for normal somatic cell division), targeting KIF18A It is possible to significantly reduce its toxicity and help improve the therapeutic safety window of tubulin-targeted drugs in clinical practice.
  • KIF18A protein is highly expressed in a variety of tumors, including colon cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer and ovarian cancer.
  • KIF18A plays a key role in the occurrence, development and metastasis of breast cancer, and its high expression predicts poor prognosis of patients.
  • KIF18A is required for proliferation of cells with chromosomal instability derived from triple-negative breast cancer or colorectal cancer, but is not required in diploid cells. Knocking out the KIF18A gene resulted in infertility in male mice, but not in female mice.
  • KIF18A mRNA expression was significantly associated with higher tumor grade and larger tumors in breast cancer patients, and KIF18A was an independent predictor of breast cancer lymph node metastasis, with a risk coefficient of 3.2.
  • inhibiting the expression of KIF18A not only affects the key function of KIF18A in cell mitosis, but also reduces cancer cell migration by stabilizing leading microtubules, ultimately leading to the inactivation of the PI3K-AKT signaling pathway and inducing cell apoptosis.
  • KIF18A protein inhibitors may be a new breakthrough in cancer drugs.
  • the present invention provides a compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope label, solvate, polymorph, pharmaceutical Acceptable salts or prodrug compounds thereof:
  • A is selected from fused ring groups that are unsubstituted or optionally substituted by one, two or more R a ;
  • the fused ring groups include two, three or four independently selected from saturated or Partially unsaturated C 3-14 carbocyclic ring, C 6-14 aromatic ring, 5-14 membered heteroaromatic ring, 3-14 membered heterocyclic ring;
  • Each R a is the same or different, independently selected from H, OH, halogen, cyano, NH 2 , NO 2 , the following groups that are unsubstituted or optionally substituted by one, two or more R a1 : C 1-12 alkyl, C 1-12 alkoxy, C 3-12 cycloalkyl; or, two R a connected to the same ring carbon atom and the carbon atom to which they are connected together form a saturated or partially unsaturated group.
  • each R a1 is the same or different, independently selected from H , OH, halogen, cyano, NH 2 , NO 2 , C 1-12 alkyl, C 1-12 alkoxy, C 3-12 cycloalkyl;
  • Each R e1 , Re2 , Re3 , Re4 , Re5 , Re6 , and Re7 are the same or different, and are independently selected from H, C 1-12 alkyl, hydroxy C 1-12 alkyl, halo C 1-12 alkyl, halo C 1-12 alkoxy, cyano C 1-12 alkyl, cyano C 1-12 alkoxy, C 3-12 cycloalkyl, 3-14 membered heterocycle Base, C 1-12 alkoxy-C 1-12 alkyl;
  • M is selected from C 3-12 cycloalkyl, C 3-12 cycloalkenyl or nitrogen-containing 3-14 membered heterocyclyl which is unsubstituted or optionally substituted by one, two or more R m ; each R m are the same or different, and are independently selected from H, halogen, cyano, C 1-12 alkyl, halo C 1-12 alkyl, cyano C 1-12 alkyl, C 1-12 alkoxy , cyano C 1-12 alkoxy group; or, two R m connected to the same ring carbon atom and the carbon atom to which they are connected together form a saturated or partially unsaturated C 3-14 carbon ring;
  • Y 1 , Y 2 , and Y 3 are the same or different, and are independently selected from N or CH.
  • A is selected from fused ring groups that are unsubstituted or optionally substituted with one, two or more Ra ; said fused ring groups comprise two, three or four independently selected from each other.
  • Each R a is the same or different, and is independently selected from H, OH, halogen, cyano, NH 2 , NO 2 , C 1-6 alkyl, C 1-6 alkoxy, C 3-8 cycloalkyl , Halogenated C 1-6 alkyl, Halogenated C 1-6 alkoxy, Cyano C 1-6 alkyl, Cyano C 1-6 alkoxy, C 3-8 cycloalkyl -C 1- 6 alkoxy; or, two R a connected to the same ring carbon atom and the carbon atom to which it is connected together form a saturated or partially unsaturated C 3-8 carbocyclic ring; or, two non-adjacent R a is connected with its end group to form a C 1-3 alkylene group;
  • A is selected from fused ring groups that are unsubstituted or optionally substituted by one, two or more Ra ; said fused ring groups are selected from,
  • T is selected from CH 2 , CH, NH, NR a or O;
  • Z is selected from CH 2 , CH, NH, NR a or O;
  • X is selected from N or CH
  • p and q are independently selected from 0, 1, 2, and 3, and p and q are not 0 at the same time;
  • r and s are independently selected from 0, 1, 2, and 3, and r and s are not 0 at the same time.
  • A is selected from fused ring groups that are unsubstituted or optionally substituted with one, two or more R a ; said fused ring groups are selected from
  • Z is selected from CH 2 , CH, NH, NR a or O;
  • X is selected from N or CH
  • r and s are independently selected from 0, 1, 2, and 3, and r and s are not 0 at the same time.
  • A is selected from
  • T is selected from CH 2 , CH, NH, NR a or O;
  • Z is selected from CH 2 , CH, NH, NR a or O;
  • X is selected from N or CH
  • n is selected from 0, 1, 2, 3, 4 or 5;
  • p and q are independently selected from 0, 1, 2, and 3, and p and q are not 0 at the same time;
  • r and s are independently selected from 0, 1, 2, and 3, and r and s are not 0 at the same time.
  • A is selected from fused ring groups that are unsubstituted or optionally substituted by one, two or more R; the fused ring group consists of two, three, four or more Selected from benzene ring, pyridine ring, imidazole ring, piperazine ring, piperidine ring, tetrahydropyrrole ring, tetrahydropyran ring, tetrahydrofuran ring, furan ring, morpholine ring, cyclopropane ring, cyclobutane ring, ring It is formed by the fusion of pentane ring, cyclohexane ring, cycloheptane ring and cycloheximine ring.
  • A is selected from unsubstituted or optionally substituted with one, two or more Ra
  • each R a is the same or different and is independently selected from the group consisting of H, OH, F, Cl, cyano, methyl, ethyl, isopropyl, methoxy, ethoxy, trifluoromethyl group, difluoromethoxy, trifluoromethoxy, cyclopropyl, cyclopropylmethoxy; or, two R a connected to the same ring carbon atom and the carbon atom to which they are connected together form cyclopropane Ring, cyclobutane ring, cyclopentane ring, cyclohexane ring, cycloheptane ring; or, two non-adjacent R a are connected with their end groups to form a methylene or ethylene group together.
  • A is selected from
  • E is selected from
  • M is selected from C 3-8 cycloalkyl, C 3-8 cycloalkenyl or nitrogen-containing 3-8 membered hetero, which is unsubstituted or optionally substituted by one, two or more R m Ring group; each R m is the same or different, independently selected from H, halogen, cyano, C 1-6 alkyl, halo C 1-6 alkyl, cyano C 1-6 alkyl, C 1 -6 alkoxy, cyano C 1-6 alkoxy; alternatively, two R m connected to the same ring carbon atom and the carbon atom to which they are connected together form a saturated or partially unsaturated C 3-8 carbon ring;
  • M is selected from
  • Y 1 , Y 2 , and Y 3 are not N at the same time;
  • Y 1 when Y 1 is CH, Y 2 is CH, Y 3 is N or CH; when Y 1 is CH, Y 2 is N, Y 3 is N or CH; when Y 1 is N, Y 2 is CH, Y 3 is N or CH; when Y 1 is N, Y 2 is N, Y 3 is CH.
  • the compound represented by formula (I) has the following structure:
  • A, M, E, Y 1 , Y 2 , Y 3 independently of each other have the definitions set forth herein.
  • the compound represented by formula (I) has the following structure:
  • M, E, Y 1 , Y 2 , Y 3 , Ra , X, Z, T, n, p, q, r, s independently of each other have the definitions set forth herein.
  • the compound represented by formula (I) has the following structure:
  • A, Y 1 , Y 2 and Y 3 independently have the definitions described herein.
  • the compound represented by formula (I) has the following structure:
  • Y 1 , Y 2 , Y 3 , R a , X, Z, T, n, p, q, r, s independently of each other have the definitions set forth herein.
  • the compound represented by formula (I) has the following structure:
  • Y 1 , Y 2 , Y 3 , R a , X, Z, n, r, s independently of each other have the definitions set forth herein.
  • the compound represented by formula (I) is selected from the following structures:
  • the present invention also provides a preparation method of the compound represented by formula (I), which includes the following steps:
  • A, E, M, Y 1 , Y 2 and Y 3 independently have the definitions mentioned above; L is selected from halogen, such as Cl, Br, I.
  • the present invention also provides a pharmaceutical composition, which contains a therapeutically effective amount of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope label, solvate, polymorph At least one of a compound, a pharmaceutically acceptable salt or a prodrug compound thereof.
  • the pharmaceutical composition further includes one or more pharmaceutically acceptable excipients.
  • the pharmaceutical composition may further contain one or more additional therapeutic agents.
  • the present invention also provides a method for treating tumor diseases, which includes administering to a patient a prophylactically or therapeutically effective amount of the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope label, At least one of a solvate, a polymorph, a pharmaceutically acceptable salt or a prodrug compound thereof.
  • the present invention also provides a method for treating tumor diseases, which includes administering to a patient a prophylactically or therapeutically effective amount of the above-mentioned pharmaceutical composition.
  • the neoplastic diseases include intestinal cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer and ovarian cancer.
  • the patient includes a mammal, preferably a human.
  • the present invention also provides compounds represented by formula (I) for treating tumor diseases, their racemates, stereoisomers, tautomers, isotope labels, solvates, polymorphs, and pharmaceutically acceptable compounds. At least one of the accepted salts or prodrug compounds thereof, or a pharmaceutical composition thereof.
  • the present invention also provides the compound represented by formula (I), its racemate, stereoisomer, tautomer, isotope label, solvate, polymorph, pharmaceutically acceptable salt or its precursor. Use of at least one of the pharmaceutical compounds in the preparation of a medicament.
  • the use may be the use in the preparation of a medicament for treating KIF18A-mediated conditions and/or diseases, such as the use in the preparation of a KIF18A inhibitor medicament.
  • the disease is, for example, cancer, including bowel cancer, breast cancer, lung cancer, pancreatic cancer, prostate cancer, bladder cancer, head and neck cancer, cervical cancer or ovarian cancer.
  • the present invention creatively obtains a class of structurally novel compounds, which not only have good KIF18A inhibitory effect and OVCAR-3 in vitro cell activity, but also have significantly improved physical and chemical properties (solubility, permeability), and OVCAR-3 can be used as medicine in vivo.
  • the efficiency is significantly improved and the safety is good.
  • Such compounds can be used to treat KIF18A-mediated conditions and/or diseases, such as tumor diseases, and to prepare medicaments for treating such conditions or diseases.
  • C 1-12 alkyl is understood to mean straight-chain and branched alkyl groups having 1 to 12 carbon atoms
  • C 1-8 alkyl means having 1, 2, 3, 4, 5, 6 , 7, or 8 carbon atoms in straight-chain and branched-chain alkyl groups
  • C 1-6 alkyl means straight-chain and branched-chain alkyl groups with 1, 2, 3, 4, 5 or 6 carbon atoms.
  • the alkyl group is, for example, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, 2-methylbutyl, 1-Methylbutyl, 1-ethylpropyl, 1,2-dimethylpropyl, neopentyl, 1,1-dimethylpropyl, 4-methylpentyl, 3-methylpentyl base, 2-methylpentyl, 1-methylpentyl, 2-ethylbutyl, 1-ethylbutyl, 3,3-dimethylbutyl, 2,2-dimethylbutyl, 1,1-dimethylbutyl, 2,3-dimethylbutyl, 1,3-dimethylbutyl or 1,2-dimethylbutyl, etc. or their isomers.
  • C 3-12 cycloalkyl is understood to mean a saturated monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spirocyclic) hydrocarbon ring or tricycloalkane, which has 3 to 12 carbon atoms, "C 3-10 cycloalkyl” is preferred, and “C 3-8 cycloalkyl” is more preferred.
  • C 3-12 cycloalkyl is understood to mean a saturated monovalent monocyclic, bicyclic (such as bridged, spiro) hydrocarbon ring or tricycloalkane having 3, 4, 5, 6, 7, 8 , 9, 10, 11 or 12 carbon atoms.
  • the C 3-12 cycloalkyl group may be a monocyclic hydrocarbon group, such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecanyl, or bicyclic Hydrocarbon groups such as bornyl, indolyl, hexahydroindolyl, tetrahydronaphthyl, decahydronaphthyl, bicyclo[2.1.1]hexyl, bicyclo[2.2.1]heptyl, bicyclo[2.2.
  • a monocyclic hydrocarbon group such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl or cyclodecanyl, or bicyclic Hydrocarbon groups such as borny
  • C 3-12 cycloalkenyl should be understood to mean a monovalent monocyclic, bicyclic (such as fused ring, bridged ring, spirocyclic ring) or tricyclic alkene containing a carbon-carbon double bond, which has 3 to 12 carbon atoms. atoms, preferably "C 3-10 cycloalkenyl", more preferably “C 3-8 cycloalkenyl”, which may have 3, 4, 5, 6, 7, 8, 9, 10, 11 or 12 carbon atoms .
  • the C 3-12 cycloalkenyl group may be a monocyclic hydrocarbon group, such as cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclohexenyl, cycloheptenyl, cyclooctenyl, cyclononenyl or Cyclodecenyl, or bicyclic hydrocarbon radical such as spiro[2.5]oct-5-enyl, spiro[3.5]non-6-enyl, spiro[4.5]dec-7-enyl.
  • C 6-14 aryl is understood to mean preferably a monovalent aromatic or partially aromatic monocyclic, bicyclic or tricyclic hydrocarbon ring having 6 to 14 carbon atoms, which may be a monoaromatic ring or Polyaromatic rings fused together are preferably "C 6-10 aryl”.
  • C 6-14 aryl is understood to mean preferably a monovalent or partially aromatic monocyclic, bicyclic or Tricyclic hydrocarbon rings (“C 6-14 aryl”), especially rings with 6 carbon atoms (“C 6 aryl”), such as phenyl; or biphenyl, or with 9 carbon atoms a ring (“C 9 aryl”), such as indanyl or indenyl, or a ring having 10 carbon atoms (“C 10 aryl”), such as tetrahydronaphthyl, dihydronaphthyl or naphthyl, Either a ring with 13 carbon atoms (“C 13 aryl”), such as fluorenyl, or a ring with 14 carbon atoms (“C 14 aryl”), such as anthracenyl.
  • C 6-20 aryl group When the C 6-20 aryl group is substituted, it may be mono- or poly-substituted. Moreover, there is no restriction on the substitution
  • 5-14 membered heteroaryl is understood to include monovalent monocyclic, bicyclic (such as fused, bridged, spiro) or tricyclic aromatic ring systems having 5 to 14 ring atoms And contain 1-5 heteroatoms independently selected from N, O and S, such as "5-10 membered heteroaryl".
  • the term "5-14 membered heteroaryl” is understood to include monovalent monocyclic, bicyclic or tricyclic aromatic ring systems having 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 ring atoms, in particular 5 or 6 or 9 or 10 carbon atoms, and it contains 1 to 5, preferably 1 to 3 heteroatoms each independently selected from N, O and S and, additionally in each case The following can be benzo-fused.
  • Heteroaryl also refers to a group in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic or heterocyclyl rings, wherein the radical or point of attachment is on the heteroaromatic ring.
  • Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7- or 8-indolizinyl, 1-, 3-, 4-, 5-, 6- or 7-isoindolyl , 2-, 3-, 4-, 5-, 6- or 7-indolyl, 2-, 3-, 4-, 5-, 6- or 7-indazolyl, 2-, 4-, 5 -, 6-, 7- or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8- or 9-quinolinyl, 2-, 3-, 4-, 5 -, 6-, 7- or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl, 1-, 4-, 5-, 6- , 7- or 8-phthalazinyl, 2-, 3-, 4-, 5- or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7- or 8-quinazole Phylyl, 3-, 4-, 5-, 6-, 7- or 8-cinnolinyl, 2-,
  • Typical fused heteroaryl groups include, but are not limited to, 2-, 3-, 4-, 5-, 6-, 7- or 8-quinolyl, 1-, 3-, 4-, 5-, 6-, 7- or 8-isoquinolyl, 2-, 3-, 4-, 5-, 6- or 7-indolyl, 2-, 3-, 4-, 5-, 6- or 7-benzo [b]Thienyl, 2-, 4-, 5-, 6- or 7-benzoxazolyl, 2-, 4-, 5-, 6- or 7-benzimidazolyl and 2-, 4-, 5-, 6- or 7-benzothiazolyl.
  • the carbon atom on the 5-14-membered heteroaryl ring can be connected to other groups, or it can be a 5-14-membered heteroaryl group.
  • Heteroatoms on the aryl ring are attached to other groups.
  • the 5-14 membered heteroaryl group is substituted, it may be mono- or poly-substituted.
  • there is no restriction on the substitution position for example, the hydrogen bonded to the carbon atom on the heteroaryl ring may be substituted, or the hydrogen bonded to the heteroatom on the heteroaryl ring may be substituted.
  • 3-14 membered heterocyclyl refers to a saturated or unsaturated non-aromatic ring or ring system, for example, which is a 4-, 5-, 6- or 7-membered monocyclic ring or ring system.
  • the heterocyclyl group may be selected from "3-10 membered heterocyclyl group".
  • heterocyclyl means a saturated or unsaturated non-aromatic ring or ring system and containing at least one heteroatom selected from O, S and N.
  • the heterocyclyl group may be attached to the remainder of the molecule through any of the carbon atoms or a nitrogen atom, if present.
  • the heterocyclyl group may include fused or bridged rings as well as spirocyclic rings.
  • the heterocyclyl group may include, but is not limited to: 4-membered rings, such as azetidinyl, oxetanyl; 5-membered rings, such as tetrahydrofuranyl, dioxolyl, pyrrole Alkyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl; or 6-membered ring, such as tetrahydropyranyl, piperidinyl, morpholinyl, dithianyl, thiomorpholinyl, piperazinyl Or trithialkyl; or 7-membered ring, such as diazacycloheptyl.
  • 4-membered rings such as azetidinyl, oxetanyl
  • 5-membered rings such as tetrahydrofuranyl, dioxolyl, pyrrole Alkyl, imidazolidinyl, pyrazolidinyl, pyrrolinyl
  • the heterocyclyl group may be benzo-fused.
  • the heterocyclyl group may be bicyclic, such as but not limited to a 5,5-membered ring, such as a hexahydrocyclopenta[c]pyrrole-2(1H)-yl ring, or a 5,6-membered bicyclic ring, such as a hexahydropyrrole And [1,2-a]pyrazine-2(1H)-yl ring.
  • Heterocyclyl may be partially unsaturated, i.e.
  • the carbon atom on the 3-14-membered heterocyclic group can be connected to other groups, or it can be a 3-14-membered heterocyclic group.
  • the heterocyclic atoms on the ring are connected to other groups.
  • the nitrogen atom on the piperazinyl may be connected to other groups.
  • the nitrogen atom on the piperidinyl ring and the carbon atom in the para position may be connected to other groups.
  • spiro ring refers to a ring system in which two rings share one ring-forming atom.
  • fused ring refers to a ring system in which two rings share 2 ring-forming atoms.
  • bridged ring refers to a ring system in which two rings share more than three ring-forming atoms.
  • halogen means fluorine, chlorine, bromine and iodine.
  • Halo means substituted with one or more halogens.
  • R a described herein is a substituent on the fused ring group A, which means that R a can be optionally substituted at any position of the fused ring group A, as shown in the structural formula means that R a can be in the ring
  • the N-containing heterocycle on the left is substituted, which also means that R a can be in the ring
  • the ring containing X on the right side is substituted, and the total number of substitutions of R a on the fused ring group A is n.
  • C 1-12 alkyl also applies to C 1-12 alkyloxy (i.e., C 1-12 alkoxy) .
  • alkylene refers to a divalent group as defined above, for example the term “alkylene” refers to a divalent alkyl group, wherein the alkyl group is as defined above As defined, the alkylene group is preferably an alkylene group having 1 to 12 carbon atoms (i.e., C 1-12 alkylene), and more preferably an alkylene group containing 1 to 6 carbon atoms (i.e. , C 1-6 alkylene). base).
  • C 3-14 carbocyclic ring refers to a saturated or unsaturated aliphatic hydrocarbon ring with 3 to 14 carbon atoms, including cycloalkane ring (C 3-14 cycloalkane ring) with 3 to 14 carbon atoms, A cycloalkene ring of 3 to 14 carbon atoms (C 3-12 cycloalkene ring) or a cycloalkyne ring of 3 to 14 carbon atoms (C 3-12 cycloalkyne ring).
  • the compound represented by formula (I) may exist in the form of various pharmaceutically acceptable salts. If these compounds have a basic center, they can form acid addition salts; if these compounds have an acidic center, they can form base addition salts; if these compounds contain both an acidic center (such as a carboxyl group) and a basic center (such as amino), which can also form internal salts.
  • the compounds of the invention may exist in the form of solvates (eg hydrates), wherein the compounds of the invention comprise as structural elements of the crystal lattice of said compounds a polar solvent, in particular such as water, methanol or ethanol.
  • a polar solvent in particular such as water, methanol or ethanol.
  • the amount of polar solvent, especially water, may be present in stoichiometric or non-stoichiometric ratios.
  • the compounds of the invention may be chiral and therefore may exist in various enantiomeric forms. These compounds may thus exist in racemic or optically active form.
  • the compounds of the present invention cover isomers in which each chiral carbon is in the R or S configuration, or their mixtures and racemates.
  • the compounds of the invention or intermediates thereof can be separated into enantiomeric compounds by chemical or physical methods known to those skilled in the art, or used in synthesis in this form. In the case of racemic amines, the diastereomers are prepared from the mixture by reaction with optically active resolving reagents.
  • suitable resolving agents are optically active acids, such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, appropriately N-protected amino acids (e.g. N- Benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids.
  • optically active acids such as the R and S forms of tartaric acid, diacetyltartaric acid, dibenzoyltartaric acid, mandelic acid, malic acid, lactic acid, appropriately N-protected amino acids (e.g. N- Benzoylproline or N-phenylsulfonylproline) or various optically active camphorsulfonic acids.
  • Suitable eluents for this purpose are aqueous or alcoholic solvent mixtures, for example, hexane/isopropanol/acetonitrile.
  • the corresponding stable isomers can be separated according to known methods, for example by extraction, filtration or column chromatography.
  • patient refers to any animal including mammals, preferably mice, rats, other rodents, rabbits, dogs, cats, pigs, cattle, sheep, horses or primates, most preferably humans.
  • terapéuticaally effective amount refers to the amount of an active compound or drug that a researcher, veterinarian, physician, or other clinician is seeking to elicit a biological or medical response in a tissue, system, animal, individual, or human, and includes one of the following or more of: (1) Prevention of disease: e.g., prevention of a disease, disorder, or condition in an individual who is susceptible to the disease, disorder, or condition but who has not yet experienced or developed the pathology or symptoms of the disease. (2) Inhibition of disease: e.g., inhibition of a disease, disorder, or condition (i.e., preventing further progression of pathology and/or symptoms) in an individual who is experiencing or developing pathology or symptoms of the disease, disorder, or condition.
  • Prevention of disease e.g., prevention of a disease, disorder, or condition in an individual who is susceptible to the disease, disorder, or condition but who has not yet experienced or developed the pathology or symptoms of the disease.
  • Inhibition of disease e.g., inhibition of a disease, disorder
  • Disease amelioration e.g., alleviation of a disease, disorder, or condition (i.e., reversal of the pathology and/or symptoms) in an individual who is experiencing or developing the pathology or symptoms of the disease, disorder, or condition.
  • the first step is the synthesis of 6-methoxynaphthalene-1-amine (compound 1-2):
  • hydroxylamine hydrochloride (206.2mg, 2.967mmol, 1.5eq) to a solution of 6-methoxynaphthalene-1-carboxylic acid (400mg, 1.978mmol, 1eq) in polyphosphoric acid (2mL) at room temperature, and stir at 80 degrees Celsius. Reaction takes 2 hours. The reaction mixture was quenched with ice water (30 mL) at 0°C. The aqueous phase was extracted with ethyl acetate (1 ⁇ 50 mL). The aqueous phase was then basified to pH ⁇ 9 with solid sodium hydroxide, and extracted with ethyl acetate (3 ⁇ 100 mL).
  • the crude product was purified by high performance liquid phase under the following conditions (chromatography column specifications: XBridge Prep OBD C18 Column, 30 )--HPLC--merk; flow rate: 60mL/min; elution gradient: 40%B to 58%B, 10min, 58%B; detection wavelength: UV 220nm; retention time (minutes): 8.23), get 2- ⁇ 6-Azaspiro[2.5]octane-6-yl ⁇ -4-(2-hydroxyethanesulfonylamino)-N-(6-methoxynaphthalen-1-yl)benzamide (compound 1, 30.32 mg, 38.11%).
  • the first step is the synthesis of 4-(4-bromo-1H-benzimidazol-2-yl)butan-1-ol (compound 21-3):
  • the second step is the synthesis of 9-bromo-benzo[4,5]imidazo[1,2-a]piperidine (compound 21-4):
  • the third step is the synthesis of benzo[4,5]imidazo[1,2-a]piperidin-9-ylcarbamic acid tert-butyl ester (compound 21-6):
  • the fourth step is the synthesis of benzo[4,5]imidazo[1,2-a]piperidin-9-amine (compound 21-7):
  • potassium phosphate was added to a solution of benzo[4,5]imidazo[1,2-a]piperidin-9-amine (240.73mg, 0.641mmol, 1.2eq) in dichloromethane (2mL) at room temperature. (340.08mg, 1.602mmol, 3eq), after stirring for 1 minute, 4-iodo-2-(6-azaspiro[2.5]octane-6-yl)benzoyl chloride (100mg, 0.534mmol, was added at room temperature, 1eq) and a solution of N,N-diisopropylethylamine (207.08mg, 1.602mmol, 3eq) in dichloromethane (1mL).
  • the reaction solution was allowed to react at room temperature for 2 hours.
  • the reaction mixture was quenched with water (20 mL) at room temperature.
  • the reaction mixture was extracted with dichloromethane (3 x 20 mL).
  • the organic phases were combined, backwashed with saturated brine (1 ⁇ 30 mL), and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The resulting mixture was used without further purification.
  • Step 6 4-((2-hydroxyethyl)sulfonylamino)-2-(6-azaspiro[2.5]octane-6-yl)-N-(benzo[4,5]imidazo[ Synthesis of 1,2-a]piperidin-9-yl)benzamide (compound 21):
  • the crude product was purified by high performance liquid phase under the following conditions (chromatography column specifications: Kinetex EVO C18 Column, 30*150, 5um; mobile phase A: water (10mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/ min; elution gradient: 10% B to 50% B, 8 min, 50% B; detection wavelength: UV 220nm; retention time (minutes): 7.47), obtain 4-((2-hydroxyethyl)sulfonylamino) -2-(6-azaspiro[2.5]octane-6-yl)-N-(benzo[4,5]imidazo[1,2-a]piperidin-9-yl)benzamide ( Compound 21, 14.1 mg, 17.63%).
  • the first step is the synthesis of (Z)-N-(2,6-dibromophenyl)piperidine-2-imine (compound 22-3):
  • the second step is the synthesis of 6-bromo-benzo[4,5]imidazo[1,2-a]piperidine (compound 22-4):
  • the third step is the synthesis of (benzo[4,5]imidazo[1,2-a]piperidin-6-yl)carbamic acid tert-butyl ester (compound 22-5):
  • reaction solution was cooled to room temperature, diluted with water (20mL), and the reaction mixture was extracted with ethyl acetate (3 ⁇ 20mL). Combine the organic phases and backwash with saturated brine (1 ⁇ 20mL). Dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure.
  • the resulting residue is purified by silica gel column chromatography, petroleum ether/ethyl acetate (2:1), to obtain (benzo[4,5]imidazo[1,2 -a]piperidin-6-yl)carbamate tert-butyl ester (compound 22-5, 490 mg, 85.64%).
  • the fourth step is the synthesis of benzo[4,5]imidazo[1,2-a]piperidin-6-amine (compound 22-6):
  • reaction solution was cooled to room temperature, diluted with water (10 mL), and the reaction mixture was extracted with ethyl acetate (3 ⁇ 10 mL). The organic phases were combined, backwashed with saturated brine (1 ⁇ 10 mL), and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure.
  • the crude product was purified by high performance liquid phase under the following conditions (chromatography column specifications: Xselect CSH C18 OBD Column 30*150mm 5 ⁇ m, n; mobile phase A: water (0.1% formic acid), mobile phase B: acetonitrile; flow rate: 60mL/min; Elution gradient: 10% B to 37% B, 10 min, 37% B; detection wavelength: 220 nm; retention time (minutes): 9.03; number of runs: 0).
  • the first step is the synthesis of (2Z)-N-(2,6-dibromophenyl)pyrrolidine-2-imine (compound 36-2):
  • the second step is the synthesis of 5-bromo-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (compound 36-3):
  • reaction solution was cooled to room temperature, quenched by adding water (80 mL), and extracted with ethyl acetate (3 ⁇ 60 mL). The organic phases were combined, backwashed with saturated brine (1 ⁇ 80 mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure. 5-Bromo-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (2 g, 74.07%) was obtained.
  • the third step is the synthesis of (2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamic acid tert-butyl ester (compound 36-5):
  • reaction solution was raised to 100°C, and the reaction solution was stirred and reacted for 1 hour.
  • the reaction solution was cooled to room temperature and quenched by adding water (80 mL). And extracted with ethyl acetate (3 ⁇ 60mL). The organic phases were combined, backwashed with saturated brine (1 ⁇ 80 mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure.
  • the fourth step is the synthesis of 2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole-5-amine (compound 36-6):
  • reaction solution was cooled to room temperature, quenched by adding water, and the reaction mixture was extracted with ethyl acetate (3 ⁇ 10 mL). The organic phases were combined, backwashed with saturated saline solution (1 ⁇ 10 mL), and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure.
  • the crude product obtained was purified by preparative high-pressure liquid phase under the following conditions: chromatography column specifications: Kinetex EVO C18 column, 30*150, 5 ⁇ m; mobile phase A: water (10mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; Flow rate: 60mL/min; elution gradient: 30%B to 65%B in 8min, 60%B; detection wavelength: UV 220nm; retention time (minutes): 6.12, obtain N-(2,3-dihydro-1H -Benzo[d]pyrrolo[1,2-a]imidazol-5-yl)-4-(2-hydroxyethanesulfonylamino)-2-(6-azaspiro[2.5]octane-6- base) benzamide (4.22 mg, 6.60%).
  • the first step is the synthesis of 5'-bromo-3',4'-dihydrospiro[cyclopentane-1,2'-naphthalene]-1'-one (compound 51-3):
  • the second step is the synthesis of 5'-bromo-3',4'-dihydro-1'H-spiro[cyclopentane-1,2'-naphthalene] (compound 51-4):
  • the third step is the synthesis of ⁇ 3',4'-dihydro-1'H-spiro[cyclopentane-1,2'-naphthalene]-5'-yl ⁇ carbamic acid tert-butyl ester (compound 51-5):
  • the fourth step is the synthesis of 3',4'-dihydro-1'H-spiro[cyclopentane-1,2'-naphthalene]-5'-amine (compound 51-6):
  • reaction solution was stirred at room temperature for 1 hour.
  • the reaction mixture was extracted with ethyl acetate (3 ⁇ 20 mL).
  • the organic phases were combined, backwashed with saturated saline solution (1 ⁇ 20 mL), and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (10:1), to obtain 2- ⁇ 6-azaspiro[2.5]octane-6-yl ⁇ -N- ⁇ 3',4' -Dihydro-1'H-spiro[cyclopentane-1,2'-naphthalene]-5'-yl ⁇ -4-iodobenzamide (100 mg, 90.34%).
  • the reaction solution was heated to 120°C and stirred for 1 hour.
  • the reaction solution was cooled to room temperature, quenched by adding water, and the reaction mixture was extracted with ethyl acetate (3 ⁇ 10 mL).
  • the organic phases were combined, backwashed with saturated saline solution (1 ⁇ 10 mL), and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure.
  • the crude product obtained was purified by preparative high-pressure liquid phase under the following conditions: chromatography column specifications: Kinetex EVO C18 column, 30*150, 5 ⁇ m; mobile phase A: water (10mmol/L ammonium bicarbonate), mobile phase B: acetonitrile; Flow rate: 60mL/min; elution gradient: 45%B to 96%B in 8min, 96%B; detection wavelength: UV 220nm; retention time (minutes): 7.62, obtaining 2- ⁇ 6-azaspiro[2.5] Octan-6-yl ⁇ -N- ⁇ 3',4'-dihydro-1'H-spiro[cyclopentane-1,2'-naphthalene]-5'-yl ⁇ -4-(2-hydroxy Ethylsulfonamido)benzamide (10 mg, 20.06%).
  • the first step is the synthesis of (2-chloroethyl)dimethylsulfonium iodide (compound 56-2):
  • the second step is the synthesis of 5'-bromo-3',4'-dihydrospiro[cyclopropane-1,2'-naphthalene]-1'-one (compound 56-4):
  • reaction solution was quenched with water and extracted with ethyl acetate (3 ⁇ 50 mL). The organic phases were combined and backwashed with saturated brine (2 ⁇ 50 mL). The resulting mixture was filtered and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (5:1), to obtain 5'-bromo-3',4'-dihydrospiro[cyclopropane-1,2'-naphthalene]-1 '-Ketone (1.8g, 80.67%).
  • the third step is the synthesis of 5'-bromo-3',4'-dihydro-1'H-spiro[cyclopropane-1,2'-naphthalene] (compound 56-5):
  • reaction solution was cooled to room temperature, filtered, the filter cake was washed with ethyl acetate, the mixture was extracted with ethyl acetate (3 ⁇ 50mL), the organic phases were combined, backwashed with saturated brine (1 ⁇ 60mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure to obtain crude product N- ⁇ 3',4'-dihydro-1'H-spiro[cyclopropane-1,2'-naphthalene]-5'-yl ⁇ -1,1-diphenyl Methylmethimine (1.1 g, crude product) was directly added to the next step without further purification.
  • the fifth step is the synthesis of 3',4'-dihydro-1'H-spiro[cyclopropane-1,2'-naphthalene]-5'-amine (compound 56-8):
  • reaction solution was heated to 80°C and reacted for 2 hours.
  • the reaction solution was cooled to room temperature, extracted with dichloromethane (3 ⁇ 20 mL), the organic phases were combined, backwashed with saturated brine (1 ⁇ 30 mL), dried with anhydrous sulfuric acid, filtered, and the filtrate was concentrated under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (3:1), to obtain 2- ⁇ 6-azaspiro[2.5]octane-6-yl ⁇ -N- ⁇ 3',4' -Dihydro-1'H-spiro[cyclopropane-1,2'-naphthalene]-5'-yl ⁇ -4-iodobenzamide (56 mg, 18.93%).
  • the first step is the synthesis of 5,5-difluoro-1-(4-methyl-3-nitropyridin-2-yl)piperidin-2-one (compound 64-3):
  • the second step is the synthesis of 8,8-difluoro-4-methyl-pyrido[2,3-d]imidazo[1,2-a]piperidine (compound 64-4):
  • the third step is the synthesis of 8,8-difluoro-4-methyl-pyrido[2,3-d]imidazo[1,2-a]piperidine-1-oxide (compound 64-5):
  • the fifth step is the synthesis of 8,8-difluoro-4-methylpyrido[2,3-d]imidazo[1,2-a]piperidin-2-amine (compound 64-7):
  • the reaction solution was heated to 60 degrees Celsius and reacted for 2 hours.
  • the reaction solution was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (3 ⁇ 10 mL).
  • the organic phases were combined, backwashed with saturated brine (2 ⁇ 10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was decompressed.
  • N-(8,8-difluoro-4-methylpyrido[2,3-d]imidazo[1,2-a]piperidin-2-yl)-4-iodo at room temperature.
  • -2-(6-Azaspiro[2.5]octane-6-yl)benzamide (30mg, 0.052mmol, 1eq)
  • 2-hydroxyethylsulfonamide (7.80mg, 0.062mmol, 1.2eq)
  • muscle Add copper iodide (0.99mg, 0.005mmol, 0.1eq) to a solution of dimethylformamide (0.8mL) of acid (0.93mg, 0.010mmol, 0.2eq) and potassium carbonate (21.54mg, 0.156mmol, 3eq).
  • the reaction solution was heated to 120 degrees Celsius and reacted for 1 hour.
  • the reaction solution was cooled to room temperature, diluted with water (10 mL), and extracted with ethyl acetate (3 ⁇ 10 mL).
  • the organic phases were combined, backwashed with saturated brine (2 ⁇ 10 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was decompressed.
  • the first step is the synthesis of (2Z)-N-(2,6-dibromophenyl)-3,3-difluoropyrrolidine-2-imine (compound 94-2):
  • the second step is the synthesis of 5-bromo-3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole (compound 94-3):
  • N,N-bis(2Z)-N-(2,6-dibromophenyl)-3,3-difluoropyrrolidine-2-imine 500mg, 1.412mmol, 1eq
  • N,N-dimethylethylenediamine (12.45mg, 0.141mmol, 0.1eq)
  • anhydrous potassium carbonate (195.21mg, 1.412mmol, 1eq)
  • copper iodide 13.45 mg, 0.071 mmol, 0.05 eq.
  • the reaction solution was heated to 100°C and stirred for 1 hour.
  • the reaction solution was cooled to room temperature, quenched by adding water, and the reaction mixture was extracted with dichloromethane (3 ⁇ 50 mL).
  • reaction solution was heated to 100°C and stirred for 1 hour. The liquid quality was monitored and the reaction was complete.
  • the reaction mixture was extracted with ethyl acetate (3 ⁇ 50mL). Combine the organic phases, backwash with saturated sodium chloride solution (1 ⁇ 50mL), and dry over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure. The resulting residue is purified by silica gel column chromatography, petroleum ether/ethyl acetate (2:1), obtaining (3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazol-5-yl)carbamic acid tert-butyl ester ( 189 mg, 72.55%).
  • the fourth step is the synthesis of 3,3-difluoro-2,3-dihydro-1H-benzo[d]pyrrolo[1,2-a]imidazole-5-amine (compound 94-5):
  • reaction solution was heated to 50°C and stirred for 1 hour.
  • the reaction solution was cooled to room temperature, diluted with water, the reaction mixture was extracted with dichloromethane (3 ⁇ 50mL), the organic phases were combined, backwashed with saturated brine (1 ⁇ 50mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was reduced to Pressure concentration.
  • reaction solution was heated to 140°C and stirred for 1 hour.
  • the reaction solution was cooled to room temperature, quenched by adding water, and the reaction mixture was extracted with ethyl acetate (3 ⁇ 30 mL). Combine the organic phases, backwash with saturated saline solution (1 ⁇ 30 mL), and dry over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid phase under the following conditions (chromatography column specifications: Kinetex EVO prep C18, 30*150, 5 ⁇ m; mobile phase A: water (10mmol/L sodium bicarbonate), mobile phase B: acetonitrile; Flow rate: 60mL/min; elution gradient: 20%B to 65%B in 8min, 65%B; detection wavelength: 220nm; retention time (min): 7.45).
  • the first step is the synthesis of (3Z)-N-(2,6-dibromophenyl)-2-azabicyclo[2.2.1]heptane-3-imine (compound 96-2):
  • the second step is the synthesis of 6-bromo-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine (compound 96-3) :
  • reaction solution was cooled to room temperature, quenched by adding water, and the reaction mixture was extracted with ethyl acetate (3 ⁇ 50 mL). The organic phases were combined, backwashed with saturated saline solution (1 ⁇ 50 mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure. 6-Bromo-1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridine (140 mg, 96.55%) was obtained.
  • reaction liquid was cooled to room temperature, and the reaction liquid was cooled to room temperature, and the reaction solution was heated to ethyl acetate (3 ⁇ 50mL) extraction. Combine the organic phases, backwash with saturated sodium chloride solution (1 ⁇ 50mL), and dry over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure. The resulting residue is purified by silica gel column chromatography, petroleum ether/acetic acid Ethyl ester (5:4), giving (1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-yl) Tert-butyl carbamate (94 mg, 63.56%).
  • Step 4 Synthesis of 1,2,3,4-tetrahydro-1,4-methylenebenzo[4,5]imidazo[1,2-a]pyridin-6-amine (compound 96-5) :
  • reaction solution was heated to 120°C and stirred for 1 hour.
  • the reaction solution was cooled to room temperature, quenched by adding water, and the reaction mixture was extracted with ethyl acetate (3 ⁇ 30 mL).
  • the organic phases were combined, backwashed with saturated saline solution (1 ⁇ 30 mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure.
  • the obtained residue was purified by preparative high performance liquid phase under the following conditions (chromatography column specifications: Kinetex EVO prep C18, 30*150, 5 ⁇ m; mobile phase A: water (10mmol/L sodium bicarbonate), mobile phase B: acetonitrile; Flow rate: 60mL/min; elution gradient: 20%B to 65%B in 8min, 65%B; detection wavelength: UV 220nm; retention time (min): 7.45).
  • the first step is the synthesis of (2Z)-N-(2,6-dibromoaniline)azepine-2-imine (compound 97-2):
  • the second step is the synthesis of 4-bromo-7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepine (compound 97-3):
  • the reaction solution was heated to 100°C and stirred for 1 hour.
  • the reaction was cooled to room temperature and extracted with ethyl acetate (3 x 50 mL).
  • the organic phases were combined, backwashed with saturated brine (1 ⁇ 50 mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (5:1), to obtain (7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2 -a]azepin-4-yl)carbamic acid tert-butyl ester (150 mg, 52.79%).
  • the fourth step is the synthesis of 7,8,9,10-tetrahydro-6H-benzo[4,5]imidazo[1,2-a]azepine-4-amine (compound 97-5):
  • reaction solution was heated to 120°C and stirred for 1 hour.
  • the reaction solution was cooled to room temperature and extracted with ethyl acetate (3 ⁇ 20 mL).
  • the organic phases were combined, backwashed with saturated saline solution (1 ⁇ 10 mL), and dried over anhydrous sodium sulfate.
  • the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the first step is the synthesis of (2Z)-N-(2,6-dibromophenyl)-3-methylpiperidin-2-imine (compound 98-2):
  • the second step is the synthesis of 6-bromo-4-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyridine (compound 98-3):
  • reaction mix The mixture was cooled to room temperature and quenched with water (10 mL). The reaction mixture was extracted with ethyl acetate (1 x 20 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (1 ⁇ 20 mL), and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was concentrated under reduced pressure. The crude product/resulting mixture was forwarded directly to the next step without further purification.
  • the reaction solution was cooled to room temperature.
  • the reaction mixture was extracted with ethyl acetate (3 ⁇ 30 mL).
  • the organic phases were combined, backwashed with saturated sodium chloride solution (1 ⁇ 40 mL), and dried over anhydrous sodium sulfate.
  • the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (5:1), to obtain (4-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1 , 2-a]pyridin-6-yl)carbamic acid tert-butyl ester (950 mg, 87.98%).
  • the fourth step is the synthesis of 4-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyridin-6-amine (compound 98-5):
  • reaction system continues at room temperature Stir for 1 hour.
  • the reaction mixture was extracted with ethyl acetate (3 ⁇ 40mL).
  • the organic phases were combined, backwashed with saturated sodium chloride solution (1 ⁇ 50mL), and dried over anhydrous sodium sulfate. After the resulting mixture was filtered, the filtrate was decompressed Concentrate.
  • Step 6 4-(2-hydroxyethanesulfonylamino)-N-(4-methyl-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyridine Synthesis of -6-yl)-2-(6-azaspiro[2.5]octane-6-yl)benzamide (compound 98):
  • reaction system is heated to 120°C and continued to stir for 1.5 hours.
  • the reaction solution was cooled to room temperature and extracted with ethyl acetate (3 ⁇ 10 mL). The organic phases were combined, backwashed with saturated sodium chloride solution (1 ⁇ 20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the crude product was purified by preparative high-performance liquid phase under the following conditions: chromatography column specifications: Kinetex 5 ⁇ m EVO C18, 30mm*150mm; mobile phase A: water (10mmol/L sodium bicarbonate), mobile phase B: acetonitrile; flow rate: 60ml/ min; elution gradient: 10%B to 37%B in 10min; detection wavelength: 254nm/220nm; retention time (min): 9.03, obtaining 4-(2-hydroxyethanesulfonylamino)-N-(4-methyl Base-1,2,3,4-tetrahydrobenzo[4,5]imidazo[1,2-a]pyridin-6-yl)-2-(6-azaspiro[2.5]octane-6 -yl)benzamide (35.6 mg, 35.39%).
  • the reaction solution was cooled to room temperature, filtered, the filter cake was washed with ethyl acetate (3 ⁇ 10 mL), and the reaction mixture was extracted with ethyl acetate (3 ⁇ 50 mL). The organic phases were combined, backwashed with saturated brine (2 ⁇ 50 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the sixth step 4-(2-hydroxyethanesulfonylamino)-N-((1S,4R)-8-methyl-1,2,3,4-tetrahydro-1,4-methylenebenzo[ Synthesis of 4,5]imidazo[1,2-a]pyridin-6-yl)-2-(6-azaspiro[2.5]octane-6-yl)benzamide (compound 99):
  • the reaction solution was heated to 120°C and stirred for 2 hours.
  • the reaction solution was cooled to room temperature, filtered, the filter cake was washed with ethyl acetate (3 ⁇ 5 mL), and the mixture was extracted with ethyl acetate (3 ⁇ 20 mL).
  • the organic phases were combined, backwashed with saturated brine (2 ⁇ 30 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the first step is the synthesis of 2-(3-bromo-2-nitrophenyl)-2-azabicyclo[2.2.2]octane-3-one (compound 100-3):
  • the reaction solution was cooled to room temperature and filtered.
  • the filter cake was washed with ethyl acetate (3 ⁇ 10mL).
  • the mixture was extracted with ethyl acetate (3 ⁇ 50mL).
  • the organic phases were combined and backwashed with saturated brine (2 ⁇ 50mL). Dry over sodium sulfate, filter, and concentrate the filtrate under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (1:1), to obtain 2-(3-bromo-2-nitrophenyl)-2-azabicyclo[2.2.2]octane. -3-one (603 mg, 52%).
  • the second step is the synthesis of 6-bromo-1,2,3,4-tetrahydro-1,4-ethylbenzo[4,5]imidazo[1,2-a]pyridine (compound 100-4):
  • reaction solution is cooled to room temperature, the reaction solution is filtered, and the filter cake is washed with ethyl acetate Wash with ester (3 ⁇ 5mL), extract the mixture with ethyl acetate (3 ⁇ 20mL), combine the organic phases, backwash with saturated brine (2 ⁇ 30mL), dry over anhydrous sodium sulfate, filter, and concentrate the filtrate under reduced pressure to obtain
  • the residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (1:1), to obtain (1,2,3,4-tetrahydro-1,4-ethylbenzo[4,5]imidazo[ 1,2-a]pyridin-6-yl)carbamic acid tert-butyl ester (90 mg, 79%).
  • the fourth step is the synthesis of 1,2,3,4-tetrahydro-1,4-ethylbenzo[4,5]imidazo[1,2-a]pyridin-6-amine (compound 100-6):
  • reaction solution was extracted with ethyl acetate (3 ⁇ 30 mL), the organic phases were combined, backwashed with saturated brine (2 ⁇ 20 mL), and dried over anhydrous sodium sulfate. Filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (3:1), to obtain 4-iodo-2-(6-azaspiro[2.5]octane-6- yl)-N-(1,2,3,4-tetrahydro-1,4-ethylbenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (110 mg ,82%).
  • Step 6 4-(2-hydroxyethanesulfonylamino)-2-(6-azaspiro[2.5]octane-6-yl)-N-(1,2,3,4-tetrahydro-1, Synthesis of 4-ethylbenzo[4,5]imidazo[1,2-a]pyridin-6-yl)benzamide (compound 100):
  • the reaction solution was stirred at 120°C for 1.5 hours.
  • the reaction solution was cooled to room temperature, and the reaction solution was filtered.
  • the filter cake was washed with ethyl acetate (3 ⁇ 5mL).
  • the mixture was extracted with ethyl acetate (3 ⁇ 30mL).
  • the organic phases were combined and backwashed with saturated brine (2 ⁇ 20mL). , dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure. The resulting residue is purified by preparative high-performance liquid phase.
  • the first step is the synthesis of methyl 4-bromo-2-(spiro[2.5]oct-5-en-6-yl)benzoate (compound 90-3):
  • 1,4-dioxane of -2- ⁇ spiro[2.5]oct-5-en-6-yl ⁇ -1,3,2-dioxaborolane (480.74 mg, 2.053 mmol, 1 eq) (8 mL) and water (1.6 mL) were added to a solution of [1,1-bis(diphenylphosphine)ferrocene]dichloride palladium dichloromethane complex (167.25 mg, 0.205 mmol, 0.1 eq).
  • the reaction solution was heated to 80°C and stirred for 1 hour.
  • the reaction solution was cooled to room temperature.
  • the reaction solution was filtered, and the filter cake was washed with ethyl acetate (3 ⁇ 50 mL).
  • the filtrate was diluted with water (30 mL). The mixture was extracted with ethyl acetate (2 ⁇ 50 mL), the organic phases were combined, backwashed with saturated brine (1 ⁇ 50 mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure. The resulting residue was purified with a reversed-phase column, and the conditions were as follows: chromatography column specifications, C18; mobile phase: acetonitrile in water, gradient 10% to 70% in 10min; detection wavelength: UV 254nm. Obtain 4-bromo-2-(spiro [2.5]oct-5-en-6-yl)benzoic acid methyl ester (320 mg, 48.52%).
  • the second step is the synthesis of 4-bromo-2- ⁇ spiro[2.5]oct-5-en-6-yl ⁇ benzoic acid (compound 90-4):
  • the reaction solution was diluted with water (15mL), the mixture was extracted with ethyl acetate (2 ⁇ 20mL), the aqueous phase was collected, and acidified to pH 4 ⁇ 5 with 1 mole per liter of dilute hydrochloric acid, and the aqueous phase was extracted with ethyl acetate (3 ⁇ 30mL) ).
  • the organic phases were combined, backwashed with saturated brine (2 ⁇ 20 mL), and dried over anhydrous sodium sulfate. Filter, and the filtrate is concentrated under reduced pressure to obtain 4-bromo-2- ⁇ spiro[2.5]oct-5-en-6-yl ⁇ benzoic acid (250 mg, 87.14%).
  • reaction solution was cooled to room temperature, extracted with ethyl acetate (3 ⁇ 30mL), the organic phases were combined, and backwashed with saturated brine (1 ⁇ 30mL). Dry over anhydrous sodium sulfate. After the resulting mixture is filtered, the filtrate is concentrated under reduced pressure.
  • the crude product is purified by high performance liquid phase under the following conditions (chromatography column specifications: Kinetex 5m EVO C18, 30mm*150mm; mobile phase A: water (10 mmol per liter ammonium bicarbonate), mobile phase B: acetonitrile; flow rate: 60 ml per minute; elution gradient: 23% B to 60% B It takes 8 minutes; detection wavelength: UV 254nm/220nm; retention time (minutes): 7.6).
  • the first step is N-(dibenzo[b,d]furan-4-yl)-4-iodo-2-(6-azaspiro[2.5]octane-6-yl)benzamide (compound 111- 2) Synthesis:
  • reaction solution was cooled to room temperature, and the reaction mixture was quenched by adding water (20 mL).
  • the reaction mixture was extracted with ethyl acetate (3 ⁇ 30 mL).
  • the organic phases were combined, backwashed with saturated brine (3 ⁇ 20 mL), and dried over anhydrous sodium sulfate.
  • the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (10:1), to obtain N-(dibenzo[b,d]furan-4-yl)-4-iodo-2-(6- Azaspiro[2.5]octan-6-yl)benzamide (140 mg, 95.73%).
  • N-(dibenzo[b,d]furan-4-yl)-4-iodo-2-(6-azaspiro[2.5]octane-6-yl)benzamide To a solution of 90 mg, 0.172 mmol, 1 eq) in N,N-dimethylformamide (2 mL), 2-hydroxyethane-1-sulfonamide (32.34 mg, 0.258 mmol, 1.5 eq) and copper iodide (8.2 mg, 0.043mmol, 0.25eq), potassium carbonate (71.43mg, 0.516mmol, 3eq) and 2-(methylamino)acetic acid (6.14mg, 0.069mmol, 0.4eq).
  • the reaction solution was heated to 120°C and stirred for 1 hour. .
  • the reaction solution was cooled to room temperature and quenched with water.
  • the reaction mixture was extracted with ethyl acetate (3 ⁇ 20 mL). The organic phases were combined, backwashed with saturated brine (2 ⁇ 20 mL), and dried over anhydrous sodium sulfate. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the crude product was purified by high performance liquid phase under the following conditions: chromatography column specifications: Kinetex EVO prep C18, 30*150, 5 ⁇ m; mobile phase A: water (10mmol/mL sodium bicarbonate), mobile phase B: acetonitrile; flow rate: 60mL/ min; elution gradient: 45% B to 70% B in 8min; detection wavelength: UV 220nm; retention time (min): 7.03.
  • chromatography column specifications Kinetex EVO prep C18, 30*150, 5 ⁇ m
  • mobile phase A water (10mmol/mL sodium bicarbonate)
  • mobile phase B acetonitrile
  • flow rate 60mL/ min
  • elution gradient 45% B to 70% B in 8min
  • detection wavelength UV 220nm
  • the first step is the synthesis of N-(7-bromoquinolin-4-yl)carbamic acid tert-butyl ester (compound 112-2):
  • the second step is the synthesis of N-(7-bromoquinolin-4-yl)carbamic acid tert-butyl ester (compound 112-4):
  • the reaction solution was heated to 80°C and stirred for one hour.
  • the reaction solution was cooled to room temperature and extracted with ethyl acetate (3 ⁇ 50 mL).
  • the organic phases were combined, backwashed with saturated brine (2 ⁇ 40 mL), and dried over anhydrous sodium sulfate.
  • the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the obtained residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (1:1), to obtain N-(7-cyclopropylquinolin-4-yl)carbamic acid tert-butyl ester (530 mg, 86.05%).
  • the third step is the synthesis of 7-cyclopropylquinolin-4-amine (compound 112-5):
  • Step 4 Preparation of 2-(6-azaspiro[2.5]octane-6-yl-N-(7-cyclopropylquinolin-4-yl)-4-iodobenzamide (compound 112-6) synthesis:
  • reaction mixture was quenched by adding water (3 mL).
  • the reaction mixture was extracted with dichloromethane (2 ⁇ 40 mL).
  • the organic phases were combined and saturated with Backwash with brine (2 ⁇ 20 mL), and dry over anhydrous sodium sulfate.
  • the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the resulting residue was purified by silica gel column chromatography, petroleum ether/ethyl acetate (5:1), to obtain 2- (6-Azaspiro[2.5]octan-6-yl-N-(7-cyclopropylquinolin-4-yl)-4-iodobenzamide (70 mg, 24.64%).
  • the reaction solution was heated to 120°C and stirred for 1.5 hours.
  • the reaction solution was cooled to room temperature and extracted with ethyl acetate (2 ⁇ 30 mL).
  • the organic phases were combined, backwashed with saturated brine (2 ⁇ 20 mL), and dried over anhydrous sodium sulfate.
  • the resulting mixture was filtered, and the filtrate was concentrated under reduced pressure.
  • the crude product was purified by high performance liquid phase to obtain 2- ⁇ 6-azaspiro[2.5]octane-6-yl ⁇ -N-(7-cyclopropylquinolin-4-yl)-4-(2-hydroxyethyl) Sulfonylamino)benzamide (18.96 mg, 31.55%).
  • NCA Imaging-Based Nuclear Count Analysis
  • the final test concentrations of AM-5308 are: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5nM.
  • Test compounds the final test concentrations are: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5nM.
  • Inhibition rate (%) 100 - (compound well reading - low reading control well reading) / (high reading control well reading - low reading control well reading) * 100
  • High reading value control well add 30nL DMSO to cells; low reading value control well: 10 ⁇ M AM-5308 well.
  • Test name KIF18A enzyme activity detection
  • AM-5308 Compound dilution and treatment: The final test concentrations of AM-5308 are: 10000, 3333.3, 1111.1, 370.3, 123.4, 41.1, 13.7, 4.5, 1.5, 0.5nM. The final test concentrations of the test compounds are: 10000, 3333.3, 1111.1, 370.3 , 123.4, 41.1, 13.7, 4.5, 1.5, 0.5nM.
  • the percent inhibition is calculated as follows:
  • Signal Ave_PC The average value of the positive control (AM-5308) on the reaction plate.
  • Signal Ave_VC The average value of the negative control (DMSO) on the reaction plate.

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Abstract

本发明提供了一种式（I）所示的化合物、药物组合物及其制备方法和应用。该化合物具有良好的KIF18A抑制作用，可以单独地或与微管形成结合复合物来调节KIF18A蛋白，用以治疗KIF18A介导的病症和/或疾病，例如肿瘤疾病，以及制备用于此类病症或疾病的药物。

Description

KIF18A抑制剂化合物、药物组合物及其制备方法和应用
本发明要求享有：
于2022年9月8日向中国国家知识产权局提交的，专利申请号为2022110982256，名称为“KIF18A抑制剂化合物、药物组合物及其制备方法和应用”的在先申请的优先权；
于2022年11月16日向中国国家知识产权局提交的，专利申请号为2022114366075，名称为“KIF18A抑制剂化合物、药物组合物及其制备方法和应用”的在先申请的优先权；
于2023年2月10日向中国国家知识产权局提交的，专利申请号为2023100987012，名称为“KIF18A抑制剂化合物、药物组合物及其制备方法和应用”的在先申请的优先权；
于2023年8月30日向中国国家知识产权局提交的，专利申请号为2023111078615，名称为“KIF18A抑制剂化合物、药物组合物及其制备方法和应用”的在先申请的优先权；
所述在先申请的全文通过引用的方式结合于本发明中。
技术领域
本发明属于医药领域，具体涉及一种KIF18A抑制剂化合物、药物组合物及其制备方法和应用。
背景技术
癌症是影响人类健康的最严重的疾病之一，死亡率和发病率常居各种疾病前列。虽然随着医疗技术和药物研发的不断发展和进步，部分患者的生活质量得到了极大的改善，但是在寻求针对不同癌症的有效治疗或治愈的药物的路途中，还有更多的未被满足的临床需求，而更多的新靶点将为未来癌症药物研发提供新的可能。
由于调控细胞周期的一个或多个基因的损伤或缺失，导致癌细胞出现不受调节的细胞增殖。各种激酶和驱动蛋白已经被鉴定在正常分裂的细胞和癌细胞的细胞周期和有丝分裂调节和进展中起关键作用。
驱动蛋白分子是以细胞内微管为轨道的动力蛋白，也称为分子马达，可以将ATP能转变为机械能，在真核细胞与细胞有丝分裂及减数分裂、组织器官的生长发育以及神经元的发育及信号传导等密切相关。驱动蛋白成员共享一个相对保守的马达结构域。根据分子中马达结构域的位置，驱动蛋白家族大致分为三类：N型驱动蛋白，即蛋白质多肽链的氨基(-NH2)末端区域具有一个马达结构域；M型驱动蛋白，即中间区域具有一个马达结构域；C型驱动蛋白，即羧基(-COOH)末端区域具有一个马达结构域。
染色体不稳定性是癌症的一个标志，由有丝分裂期间的染色体分离错误导致。靶向染色体不稳定性是药物开发中一种新兴的治疗策略。KIF18A是N型Kinesin-8驱动蛋白家族中的一员，已被证明在维持双极纺锤体的完整性和促进染色体不稳定性癌细胞的生存能力方面发挥作用。有丝分裂是一个有效的干预点，许多抗有丝分裂药物被用于临床治疗人类癌症。应用最广泛的微管蛋白抑制剂，它既可以稳定微管，也可以防止微管组装。目前抗有丝分裂的药物存在治疗窗较窄的局限性，这些问题需要开发新的靶点来解决。
尽管微管蛋白抑制剂被广泛用作治疗多种人类癌症类型的标准治疗方法，但这些药物存在对正常细胞的附带损伤，包括骨髓抑制和神经毒性。由于KIF18A在正常二倍体体细胞中可能不是必须的(KIF18A基因敲除小鼠是可存活的，但在生殖方面存在缺陷，这表明KIF18A不是正常体细胞分裂的必要基因)，因此靶向KIF18A有可能会显著降低其毒性，有利于提升临床中靶向微管蛋白药物的治疗安全窗。
KIF18A蛋白在多种肿瘤中均有高表达，包括结肠癌、乳腺癌、肺癌、胰腺癌、前列腺癌、膀胱癌、头颈部癌、宫颈癌和卵巢癌。KIF18A在乳腺癌发生、发展和转移过程中起关键作用，其高表达预示患者不良预后。KIF18A是来源于三阴性乳腺癌或结直肠癌的染色体不稳定性细胞增殖所必需的，但在二倍体细胞中不需要KIF18A。敲除KIF18A基因可导致雄性小鼠不育，但雌性小鼠没有受到影响。KIF18A mRNA表达与乳腺癌患者中较高的肿瘤等级和较大的肿瘤显著相关，且KIF18A是乳腺癌淋巴结转移的独立预测因素，风险系数为3.2。此外，抑制KIF18A表达不仅影响KIF18A在细胞有丝分裂中的关键功能，还能够通过稳定前沿微管来减少癌细胞迁移，最终导致PI3K-AKT信号通路失活而诱导细胞凋亡。
因此，对KIF18A蛋白抑制剂的开发可能是癌症药物的一个新的突破。
发明内容
为改善上述技术问题，本发明提供了一种式(I)所示的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物：
其中，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团包含两个、三个或四个彼此独立地选自饱和或部分不饱和的C3-14碳环、C6-14芳环、5-14元杂芳环、3-14元杂环的环；
每个Ra相同或不同，彼此独立地选自H、OH、卤素、氰基、NH2、NO2，无取代或任选被一个、两个或更多个Ra1取代的下列基团：C1-12烷基、C1-12烷氧基、C3-12环烷基；或者，连接在同一个环碳原子上的两个Ra与其连接的碳原子一起共同形成饱和或部分不饱和的C3-14碳环；或者，两个不相邻的Ra以其端基相连，共同形成C1-3的亚烷基；每个Ra1相同或不同，彼此独立地选自H、OH、卤素、氰基、NH2、NO2、C1-12烷基、C1- 12烷氧基、C3-12环烷基；
E选自无取代或任选被一个、两个或更多个Re取代的下列基团：-NH-S(＝O)2-Re1、-S(＝O)2-NH-Re2、-S(＝O)(＝NH)-Re3、-N(Re4)(Re5)、3-14元杂环基；每个Re相同或不同，彼此独立地选自OH、卤素、氰基、C1-12烷基、C1-12烷氧基、卤代C1-12烷基、卤代C1-12烷氧基、氰基C1-12烷基、氰基C1-12烷氧基、-N(Re6)(Re7)；
每个Re1、Re2、Re3、Re4、Re5、Re6、Re7相同或不同，彼此独立地选自H、C1-12烷基、羟基C1-12烷基、卤代C1-12烷基、卤代C1-12烷氧基、氰基C1-12烷基、氰基C1-12烷氧基、C3-12环烷基、3-14元杂环基、C1-12烷氧基-C1-12烷基；
M选自无取代或任选被一个、两个或更多个Rm取代的C3-12环烷基、C3-12环烯基或含氮的3-14元杂环基；每个Rm相同或不同，彼此独立地选自H、卤素、氰基、C1-12烷基、卤代C1-12烷基、氰基C1-12烷基、C1-12烷氧基、氰基C1-12烷氧基；或者，连接在同一个环碳原子上的两个Rm与其连接的碳原子一起共同形成饱和或部分不饱和的C3-14碳环；
Y1、Y2、Y3相同或不同，彼此独立地选自N或CH。
根据一些实施方案，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团包含两个、三个或四个彼此独立地选自饱和或部分不饱和的C3-8碳环、C6-10芳环、5-10元杂芳环、3-8元杂环的环；
每个Ra相同或不同，彼此独立地选自H、OH、卤素、氰基、NH2、NO2、C1-6烷基、C1-6烷氧基、C3-8环烷基、卤代C1-6烷基、卤代C1-6烷氧基、氰基C1-6烷基、氰基C1-6烷氧基、C3-8环烷基-C1-6烷氧基；或者，连接在同一个环碳原子上的两个Ra与其连接的碳原子一起共同形成饱和或部分不饱和的C3-8碳环；或者，两个不相邻的Ra以其端基相连，共同形成C1-3的亚烷基；
根据一些实施方案，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团选自、
其中，T选自CH2、CH、NH、NRa或O；
Z选自CH2、CH、NH、NRa或O；
X选自N或CH；
p、q彼此独立地选自0、1、2、3，且p和q不同时为0；
r、s彼此独立地选自0、1、2、3，且r和s不同时为0。
根据一些实施方案，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团选自
其中，Z选自CH2、CH、NH、NRa或O；
X选自N或CH；
r、s彼此独立地选自0、1、2、3，且r和s不同时为0。
根据一些实施方案，A选自
其中，T选自CH2、CH、NH、NRa或O；
Z选自CH2、CH、NH、NRa或O；
X选自N或CH；
n选自0、1、2、3、4或5；
p、q彼此独立地选自0、1、2、3，且p和q不同时为0；
r、s彼此独立地选自0、1、2、3，且r和s不同时为0。
根据一些实施方案，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团由两个、三个、四个或更多个选自苯环、吡啶环、咪唑环、哌嗪环、哌啶环、四氢吡咯环、四氢吡喃环、四氢呋喃环、呋喃环、吗啉环、环丙烷环、环丁烷环、环戊烷环、环己烷环、环庚烷环、环己亚胺环的环稠合而成。
根据一些实施方案，A选自无取代或任选被一个、两个或更多个Ra取代的
根据一些实施方案，每个Ra相同或不同，彼此独立地选自H、OH、F、Cl、氰基、甲基、乙基、异丙基、甲氧基、乙氧基、三氟甲基、二氟甲氧基、三氟甲氧基、环丙基、环丙基甲氧基；或者，连接在同一个环碳原子上的两个Ra与其连接的碳原子一起共同形成环丙烷环、环丁烷环、环戊烷环、环己烷环、环庚烷环；或者，两个不相邻的Ra以其端基相连，共同形成亚甲基或亚乙基。
根据一些实施方案，A选自
根据一些实施方案，E选自无取代或任选被一个或两个Re取代的下列基团：-NH-S(＝O)2-Re1、-S(＝O)2-NH-Re2、-S(＝O)(＝NH)-Re3；每个Re1、Re2、Re3相同或不同，彼此独立地选自H、C1-6烷基、羟基C1-6烷基、卤代C1- 6烷基、卤代C1-6烷氧基、氰基C1-6烷基、氰基C1-6烷氧基、C3-8环烷基、3-8元杂环基、C1-6烷氧基-C1-6烷基；每个Re相同或不同，彼此独立地选自OH、卤素、氰基、C1-6烷基、C1-6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、氰基C1-6烷基、氰基C1-6烷氧基。
根据一些实施方案，E选自
根据一些实施方案，M选自无取代或任选被一个、两个或更多个Rm取代的C3-8环烷基、C3-8环烯基或含氮的3-8元杂环基；每个Rm相同或不同，彼此独立地选自H、卤素、氰基、C1-6烷基、卤代C1-6烷基、氰基C1- 6烷基、C1-6烷氧基、氰基C1-6烷氧基；或者，连接在同一个环碳原子上的两个Rm与其连接的碳原子一起共同形成饱和或部分不饱和的C3-8碳环；
根据一些实施方案，M选自
根据一些实施方案，Y1、Y2、Y3不同时为N；
根据一些实施方案，当Y1为CH时，Y2为CH，Y3为N或CH；当Y1为CH时，Y2为N，Y3为N或CH；当Y1为N时，Y2为CH，Y3为N或CH；当Y1为N时，Y2为N，Y3为CH。
根据一些实施方案，式(I)所示的化合物具有以下结构：
其中，A、M、E、Y1、Y2、Y3彼此独立地具有本文中所述的定义。
根据一些实施方案，式(I)所示的化合物具有以下结构：
其中，M、E、Y1、Y2、Y3、Ra、X、Z、T、n、p、q、r、s彼此独立地具有本文中所述的定义。
根据一些实施方案，式(I)所示的化合物具有以下结构：
其中，A、Y1、Y2、Y3独立地具有本文中所述的定义。
根据一些实施方案，式(I)所示的化合物具有以下结构：
其中，Y1、Y2、Y3、Ra、X、Z、T、n、p、q、r、s彼此独立地具有本文中所述的定义。
根据一些实施方案，式(I)所示的化合物具有以下结构：
其中，Y1、Y2、Y3、Ra、X、Z、n、r、s彼此独立地具有本文中所述的定义。
根据一些实施方案，式(I)所示化合物选自以下结构：












本发明还提供式(I)所示化合物的制备方法，包括以下步骤：
(1)化合物a与化合物A-NH2反应得到化合物b；
(2)化合物b与化合物E-H反应得到式(I)所示化合物；
其中，A、E、M、Y1、Y2、Y3彼此独立地具有上文所述的定义；L选自卤素，如Cl、Br、I。
本发明还提供一种药物组合物，其包含治疗有效量的式(I)所示的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物中的至少一种。
根据本发明的实施方案，所述药物组合物还包括一种或多种药学上可接受的辅料。
根据本发明的实施方案，所述药物组合物还可以进一步含有一种或多种额外的治疗剂。
本发明还提供一种治疗肿瘤疾病的方法，包括给予患者预防或治疗有效量的式(I)所示的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物中的至少一种。
本发明还提供治疗肿瘤疾病的方法，包括给予患者预防或治疗有效量的上述药物组合物。
所述肿瘤疾病包括肠癌、乳腺癌、肺癌、胰腺癌、前列腺癌、膀胱癌、头颈部癌、宫颈癌和卵巢癌。
在一些实施方案中，所述患者包括哺乳动物，优选是人。
本发明还提供用于治疗肿瘤疾病的式(I)所示的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物中的至少一种，或其药物组合物。
本发明还提供式(I)所示的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物中的至少一种在制备药物中的用途。
根据本发明的实施方案，所述用途可以为在制备治疗KIF18A介导的病症和/或疾病的药物中的用途，如在制备KIF18A抑制剂药物中的用途。
根据本发明的实施方案，所述疾病例如为癌症，包括肠癌、乳腺癌、肺癌、胰腺癌、前列腺癌、膀胱癌、头颈部癌、宫颈癌或卵巢癌。
有益效果
本发明通过结构优化，创造性的获得了一类结构新颖的化合物，其不仅具有良好的KIF18A抑制作用和OVCAR-3体外细胞活性，且理化性质(溶解度、渗透性)显著提升，OVCAR-3体内药效显著提升，且安全性良好。该类化合物可用以治疗KIF18A介导的病症和/或疾病，例如肿瘤疾病，以及制备用于治疗此类病症或疾病的药物。
术语定义与说明
除非另有说明，本申请说明书和权利要求书中记载的基团和术语定义，包括其作为实例的定义、示例性的定义、优选的定义、表格中记载的定义、实施例中具体化合物的定义等，可以彼此之间任意组合和结合。这样的组合和结合后的基团定义及化合物结构，应当被理解为本申请说明书和/或权利要求书记载的范围内。
除非另有说明，本说明书和权利要求书记载的数值范围相当于至少记载了其中每一个具体的整数数值。例如，数值范围“1-14”相当于记载了数值范围“1-14”中的每一个整数数值，即1、2、3、4、5、6、7、8、9、10、11、12、13、14。
本申请通式定义中的术语“任选的”(或“任选地”、“任选”)意味着被零个、一个或多个取代基所取代的情形，例如“任选被一个、两个或更多个R取代”意味着可以不被R取代(无取代)或可以选择被一个、两个或更多个R取代。
“更多个”表示三个或三个以上。
术语“C1-12烷基”应理解为表示具有1～12个碳原子的直链和支链烷基，“C1-8烷基”表示具有1、2、3、4、5、6、7、或8个碳原子的直链和支链烷基，“C1-6烷基”表示具有1、2、3、4、5或6个碳原子的直链和支链烷基。所述烷基是例如甲基、乙基、丙基、丁基、戊基、己基、异丙基、异丁基、仲丁基、叔丁基、异戊基、2-甲基丁基、1-甲基丁基、1-乙基丙基、1,2-二甲基丙基、新戊基、1,1-二甲基丙基、4-甲基戊基、3-甲基戊基、2-甲基戊基、1-甲基戊基、2-乙基丁基、1-乙基丁基、3,3-二甲基丁基、2,2-二甲基丁基、1,1-二甲基丁基、2,3-二甲基丁基、1,3-二甲基丁基或1,2-二甲基丁基等或它们的异构体。
术语“C3-12环烷基”应理解为表示饱和的一价单环、二环(如稠环、桥环、螺环)烃环或三环烷烃，其具有3～12个碳原子，优选“C3-10环烷基”，更优选“C3-8环烷基”。术语“C3-12环烷基”应理解为表示饱和的一价单环、双环(如桥环、螺环)烃环或三环烷烃，其具有3、4、5、6、7、8、9、10、11或12个碳原子。所述C3-12环烷基可以是单环烃基，如环丙基、环丁基、环戊基、环己基、环庚基、环辛基、环壬基或环癸基，或者是双环烃基如龙脑基、吲哚基、六氢吲哚基、四氢萘基、十氢萘基、二环[2.1.1]己基、二环[2.2.1]庚基、二环[2.2.1]庚烯基、6,6-二甲基二环[3.1.1]庚基、2,6,6-三甲基二环[3.1.1]庚基、二环[2.2.2]辛基、2,7-二氮杂螺[3,5]壬烷基、2,6-二氮杂螺[3,4]辛烷基，或者是三环烃基如金刚烷基。
术语“C3-12环烯基”应理解为表示含有碳碳双键的一价单环、二环(如稠环、桥环、螺环)或三环烯烃，其具有3～12个碳原子，优选“C3-10环烯基”，更优选“C3-8环烯基”，其可以具有3、4、5、6、7、8、9、10、11或12个碳原子。所述C3-12环烯基可以是单环烃基，如环丙烯基、环丁烯基、环戊烯基、环己烯基、环庚烯基、环辛烯基、环壬烯基或环癸烯基，或者是二环烃基如螺[2.5]辛-5-烯基、螺[3.5]壬-6-烯基、螺[4.5]癸-7-烯基。
术语“C6-14芳基”应理解为优选表示具有6～14个碳原子的一价芳香性或部分芳香性的单环、二环或三环烃环，其可以是单芳族环或稠合在一起的多芳族环，优选“C6-10芳基”。术语“C6-14芳基”应理解为优选表示具有6、7、8、9、10、11、12、13或14个碳原子的一价芳香性或部分芳香性的单环、双环或三环烃环(“C6-14芳基”)，特别是具有6个碳原子的环(“C6芳基”)，例如苯基；或联苯基，或者是具有9个碳原子的环(“C9芳基”)，例如茚满基或茚基，或者是具有10个碳原子的环(“C10芳基”)，例如四氢化萘基、二氢萘基或萘基，或者是具有13个碳原子的环(“C13芳基”)，例如芴基，或者是具有14个碳原子的环(“C14芳基”)，例如蒽基。当所述C6-20芳基被取代时，其可以为单取代或者多取代。并且，对其取代位点没有限制，例如可以为邻位、对位或间位取代。
术语“5-14元杂芳基”应理解为包括这样的一价单环、二环(如稠环、桥环、螺环)或三环芳族环系：其具有5～14个环原子且包含1-5个独立选自N、O和S的杂原子，例如“5-10元杂芳基”。术语“5-14元杂芳基”应理解为包括这样的一价单环、双环或三环芳族环系：其具有5、6、7、8、9、10、11、12、13或14个环原子，特别是5或6或9或10个碳原子，且其包含1-5个，优选1-3各独立选自N、O和S的杂原子并且，另外在每一种情况下可为苯并稠合的。“杂芳基”还指其中杂芳族环与一个或多个芳基、脂环族或杂环基环稠合的基团，其中所述连接的根基或点在杂芳族环上。非限制性实例包括1-、2-、3-、5-、6-、7-或8-吲嗪基、1-、3-、4-、5-、6-或7-异吲哚基、2-、3-、4-、5-、6-或7-吲哚基、2-、3-、4-、5-、6-或7-吲唑基、2-、4-、5-、6-、7-或8-嘌呤基、1-、2-、3-、4-、6-、7-、8-或9-喹嗪基、2-、3-、4-、5-、6-、7-或8-喹啉基、1-、3-、4-、5-、6-、7-或8-异喹啉基、1-、4-、5-、6-、7-或8-酞嗪基(phthalazinyl)、2-、3-、4-、5-或6-萘啶基、2-、3-、5-、6-、7-或8-喹唑啉基、3-、4-、5-、6-、7-或8-噌啉基、2-、4-、6-或7-蝶啶基、1-、2-、3-、4-、5-、6-、7-或8-4aH咔唑基、1-、2-、3-、4-、5-、6-、7-或8-咔唑基咔唑基、1-、3-、4-、5-、6-、7-、8-或9-咔啉基、1-、2-、3-、 4-、6-、7-、8-、9-或10-菲啶基、1-、2-、3-、4-、5-、6-、7-、8-或9-吖啶基、1-、2-、4-、5-、6-、7-、8-或9-啶基、2-、3-、4-、5-、6-、8-、9-或10-菲咯啉基、1-、2-、3-、4-、6-、7-、8-或9-吩嗪基、1-、2-、3-、4-、6-、7-、8-、9-或10-吩噻嗪基、1-、2-、3-、4-、6-、7-、8-、9-或10-吩嗪基、2-、3-、4-、5-、6-或1-、3-、4-、5-、6-、7-、8-、9-或10-苯并异喹啉基、2-、3-、4-或噻吩并[2,3-b]呋喃基、2-、3-、5-、6-、7-、8-、9-、10-或11-7H-吡嗪并[2,3-c]咔唑基、2-、3-、5-、6-或7-2H-呋喃并[3,2-b]-吡喃基、2-、3-、4-、5-、7-或8-5H-吡啶并[2,3-d]-邻-嗪基、1-、3-或5-1H-吡唑并[4,3-d]-唑基、2-、4-或54H-咪唑并[4,5-d]噻唑基、3-、5-或8-吡嗪并[2,3-d]哒嗪基、2-、3-、5-或6-咪唑并[2,1-b]噻唑基、1-、3-、6-、7-、8-或9-呋喃并[3,4-c]噌啉基、1-、2-、3-、4-、5-、6-、8-、9-、10或11-4H-吡啶并[2,3-c]咔唑基、2-、3-、6-或7-咪唑并[1,2-b][1,2,4]三嗪基、7-苯并[b]噻吩基、2-、4-、5-、6-或7-苯并唑基、2-、4-、5-、6-或7-苯并咪唑基、2-、4-、4-、5-、6-或7-苯并噻唑基、1-、2-、4-、5-、6-、7-、8-或9-苯并氧杂基(benzoxapinyl)、2-、4-、5-、6-、7-或8-苯并嗪基、1-、2-、3-、5-、6-、7-、8-、9-、10-或11-4H-吡咯并[1,2-b][2]苯并氮杂基(benzazapinyl)。典型的稠合杂芳基包括但不限于2-、3-、4-、5-、6-、7-或8-喹啉基、1-、3-、4-、5-、6-、7-或8-异喹啉基、2-、3-、4-、5-、6-或7-吲哚基、2-、3-、4-、5-、6-或7-苯并[b]噻吩基、2-、4-、5-、6-或7-苯并唑基、2-、4-、5-、6-或7-苯并咪唑基和2-、4-、5-、6-或7-苯并噻唑基。当所述5-14元杂芳基与其它基团相连构成本发明的化合物时，可以为5-14元杂芳基环上的碳原子与其它基团相连，也可以为5-14元杂芳基环上的杂原子与其它基团相连。当所述5-14元杂芳基被取代时，其可以为单取代或者多取代。并且，对其取代位点没有限制，例如可以为杂芳基环上与碳原子相连的氢被取代，或者杂芳基环上与杂原子相连的氢被取代。
除非另有定义，术语“3-14元杂环基”是指饱和的或不饱和的非芳族的环或环系，例如，其是4-、5-、6-或7-元的单环、7-、8-、9-、10-、11-或12-元的二环(如稠环、桥环、螺环)或者10-、11-、12-、13-或14-元的三环环系，并且含有至少一个，例如1、2、3、4、5个或更多个选自O、S和N的杂原子，其中N和S还可以任选被氧化成各种氧化状态，以形成氮氧化物、-S(O)-或-S(O)2-的状态。优选地，所述杂环基可以选自“3-10元杂环基”。术语“3-10元杂环基”意指饱和的或不饱和的非芳族的环或环系，并且含有至少一个选自O、S和N的杂原子。所述杂环基可以通过所述碳原子中的任一个或氮原子(如果存在的话)与分子的其余部分连接。所述杂环基可以包括稠合的或桥连的环以及螺环的环。特别地，所述杂环基可以包括但不限于：4元环，如氮杂环丁烷基、氧杂环丁烷基；5元环，如四氢呋喃基、二氧杂环戊烯基、吡咯烷基、咪唑烷基、吡唑烷基、吡咯啉基；或6元环，如四氢吡喃基、哌啶基、吗啉基、二噻烷基、硫代吗啉基、哌嗪基或三噻烷基；或7元环，如二氮杂环庚烷基。任选地，所述杂环基可以是苯并稠合的。所述杂环基可以是双环的，例如但不限于5,5元环，如六氢环戊并[c]吡咯-2(1H)-基环，或者5,6元双环，如六氢吡咯并[1,2-a]吡嗪-2(1H)-基环。杂环基可以是部分不饱和的，即它可以包含一个或多个双键，例如但不限于二氢呋喃基、二氢吡喃基、2,5-二氢-1H-吡咯基、4H-[1,3,4]噻二嗪基、1,2,3,5-四氢噁唑基或4H-[1,4]噻嗪基，或者，它可以是苯并稠合的，例如但不限于二氢异喹啉基。所述3-14元杂环基与其它基团相连构成本发明的化合物时，可以为3-14元杂环基上的碳原子与其它基团相连，也可以为3-14元杂环基环上杂环原子与其它基团相连。例如当3-14元杂环基选自哌嗪基时，可以为哌嗪基上的氮原子与其它基团相连。或当3-14元杂环基选自哌啶基时，可以为哌啶基环上的氮原子和其对位上的碳原子与其它基团相连。
术语“螺环”是指两个环共用1个成环原子的环系。
术语“稠环”是指两个环共用2个成环原子的环系。
术语“桥环”是指两个环共用3个以上成环原子的环系。
术语“卤素”表示氟、氯、溴和碘。
“卤代”指被一个或多个卤素取代。
除非另有说明，本文所述的Ra为稠环基团A上的取代基，即表示Ra可以任选地在稠环基团A的任意位置上发生取代，如结构式表示Ra既可以在环左侧的含N杂环进行取代，也表示Ra可以在环右侧含X的环上取代，Ra在稠环基团A上的总取代个数为n。
除非另有说明，本文中术语的定义同样适用于包含该术语的基团，例如C1-12烷基的定义也适用于C1-12烷基氧基(即C1-12烷氧基)。
术语“亚*基”是指二价的基团，所述基团如上所定义，例如术语“亚烷基”是指二价烷基，其中烷基如上 所定义，亚烷基优选具有1至12个碳原子的亚烷基(即C1-12亚烷基)，更优选含有1至6个碳原子的亚烷基(即C1- 6亚烷基)。
术语“C3-14碳环”是指具有3至14个碳原子形成的饱和或不饱和的脂肪烃环，包括3至14个碳原子的环烷烃环(C3-14环烷烃环)、3至14个碳原子的环烯烃环(C3-12环烯烃环)或3至14个碳原子的环炔烃环(C3-12环炔烃环)。
本领域技术人员可以理解，式(I)所示化合物可以以各种药学上可接受的盐的形式存在。如果这些化合物具有碱性中心，则其可以形成酸加成盐；如果这些化合物具有酸性中心，则其可以形成碱加成盐；如果这些化合物既包含酸性中心(例如羧基)又包含碱性中心(例如氨基)，则其还可以形成内盐。
本发明的化合物可以溶剂合物(如水合物)的形式存在，其中本发明的化合物包含作为所述化合物晶格的结构要素的极性溶剂，特别是例如水、甲醇或乙醇。极性溶剂特别是水的量可以化学计量比或非化学计量比存在。
根据其分子结构，本发明的化合物可以是手性的，因此可能存在各种对映异构体形式。因而这些化合物可以以消旋体形式或光学活性形式存在。本发明的化合物涵盖了各手性碳为R或S构型的异构体或其混合物、消旋体。本发明的化合物或其中间体可以通过本领域技术人员公知的化学或物理方法分离为对映异构体化合物，或者以此形式用于合成。在外消旋的胺的情况中，通过与光学活性的拆分试剂反应，从混合物制得非对映异构体。适当的拆分试剂的示例是光学活性的酸，例如R和S形式的酒石酸、二乙酰酒石酸、二苯甲酰酒石酸、扁桃酸、苹果酸、乳酸、适当的N-保护的氨基酸(例如N-苯甲酰脯氨酸或N-苯磺酰基脯氨酸)或各种光学活性的樟脑磺酸。借助光学活性的拆分试剂(例如固定在硅胶上的二硝基苯甲酰基苯基甘氨酸、三乙酸纤维素或其它碳水化合物的衍生物或手性衍生化的异丁烯酸酯聚合物)，也可有利地进行色谱对映体拆分。用于此目的的适当的洗脱剂是含水或含醇的溶剂混合物，例如，己烷/异丙醇/乙腈。
可以根据已知的方法，例如通过萃取、过滤或柱层析来分离相应的稳定异构体。
术语“患者”是指包括哺乳动物在内的任何动物，优选小鼠、大鼠、其它啮齿类动物、兔、狗、猫、猪、牛、羊、马或灵长类动物，最优选人。
术语“治疗有效量”是指研究人员、兽医、医师或其它临床医师正在组织、系统、动物、个体或人中寻找的引起生物学或医学反应的活性化合物或药物的量，它包括以下一项或多项：(1)预防疾病：例如在易感染疾病、紊乱或病症但尚未经历或出现疾病病理或症状的个体中预防疾病、紊乱或病症。(2)抑制疾病：例如在正经历或出现疾病、紊乱或病症的病理或症状的个体中抑制疾病、紊乱或病症(即阻止病理和/或症状的进一步发展)。(3)缓解疾病：例如在正经历或出现疾病、紊乱或病症的病理或症状的个体中缓解疾病、紊乱或病症(即逆转病理和/或症状)。
具体实施方式
下文将结合具体实施例对本发明的技术方案做更进一步的详细说明。应当理解，下列实施例仅为示例性地说明和解释本发明，而不应被解释为对本发明保护范围的限制。凡基于本发明上述内容所实现的技术均涵盖在本发明旨在保护的范围内。
除非另有说明，以下实施例中使用的原料和试剂均为市售商品，或者可以通过已知方法制备。
实施例1
2-{6-氮杂螺[2.5]辛烷-6-基}-4-(2-羟基乙磺酰氨基)-N-(6-甲氧基萘-1-基)苯甲酰胺(化合物1)

第一步6-甲氧基萘-1-胺(化合物1-2)的合成：
氮气保护，室温下向6-甲氧基萘-1-甲酸(400mg,1.978mmol,1eq)的多聚磷酸(2mL)溶液中加入盐酸羟胺(206.2mg,2.967mmol,1.5eq)，80摄氏度搅拌反应2小时。反应混合物0摄氏度下用冰水(30mL)淬灭。水相用乙酸乙酯萃取(1×50mL)。再将水相用固体氢氧化钠碱化到pH～9，用乙酸乙酯萃取(3×100mL)。合并有机相，用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩，得到6-甲氧基萘-1-胺(化合物1-2，300mg,87.55％)。
MS(ESI,m/z):174.35[M+H]+，RT(min):0.760
第二步2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘-N-(6-甲氧基萘-1-基)苯甲酰胺(化合物1-4)的合成：
氮气保护，室温下向6-甲氧基萘-1-胺(150mg,0.866mmol,1eq)的二氯甲烷(3mL)溶液中加入2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(463.98mg,1.299mmol,1.5eq)，N,N,N',N'-四甲基氯甲脒六氟磷酸(971.9mg,3.464mmol,4eq)和1-甲基咪唑(711.02mg,8.660mmol,10eq)，搅拌反应2小时；液质检测到目标产物。反应混合物室温下用水(50mL)淬灭，并用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘-N-(6-甲氧基萘-1-基)苯甲酰胺(化合物1-4，300mg,67.61％)。
MS(ESI,m/z):513.35[M+H]+，RT(min):1.499
第三步2-{6-氮杂螺[2.5]辛烷-6-基}-4-(2-羟基乙磺酰氨基)-N-(6-甲氧基萘-1-基)苯甲酰胺(化合物1)的合成
在氮气保护和120摄氏度下向2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘-N-(6-甲氧基萘-1-基)苯甲酰胺(80mg,0.156mmol,1eq)的N,N-二甲基甲酰胺(0.5mL)溶液中加入2-羟基乙磺酰胺(23.45mg,0.187mmol,1.2eq)，碘化亚铜(7.43mg,0.039mmol,0.25eq)，碳酸钾(64.73mg,0.468mmol,3eq)和肌氨酸(5.56mg,0.062mmol,0.4eq)，搅拌反应过夜。加水10mL稀释，并用乙酸乙酯萃取(3×10mL)。合并有机相，用饱和食盐水反洗(3×10mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩得到粗品。粗品通过高效液相纯化，条件如下(层析柱规格:XBridge Prep OBD C18 Column,30X150mm,5μm；流动相A:水(10mmol/L碳酸氢铵)，流动相B:乙腈(0.1％二乙胺)--HPLC--merk；流速:60mL/min；洗脱梯度:40％B至58％B，10min,58％B；检测波长:UV 220nm；保留时间(分钟):8.23)，得到2-{6-氮杂螺[2.5]辛烷-6-基}-4-(2-羟基乙磺酰氨基)-N-(6-甲氧基萘-1-基)苯甲酰胺(化合物1，30.32mg,38.11％)。
MS(ESI,m/z):509.90[M+H]+，RT(min):1.572
1H NMR:(400MHz,DMSO-d6)δ11.84(s,1H),10.12(s,1H),8.07(d,1H),8.03–7.98(m,1H),7.94(d,1H), 7.66(d,1H),7.49(t,1H),7.39(d,1H),7.26(d,1H),7.22–7.16(m,1H),7.13–7.03(m,1H),4.96(s,1H),3.90(s,3H),3.78(t,2H),3.36(t,2H),3.15–2.96(m,4H),1.42(s,4H),0.26(s,4H).
使用与实施例1中类似的条件，制备了如下表中的化合物。

实施例2
2-{6-氮杂螺[2.5]辛烷-6-基}-4-(2-羟基乙磺酰氨基)-N-(6-甲氧基萘-1-基)苯甲酰胺(化合物21)

第一步4-(4-溴-1H-苯并咪唑-2-基)丁烷-1-醇(化合物21-3)的合成：
在氮气保护和100摄氏度下，将3-溴苯-1,2-二胺(2g,10.693mmol,1eq)和δ-戊内酯(4.28g,42.772mmol,4eq)的盐酸(4M,20mL)溶液搅拌反应2小时。反应液冷却至室温，混合物用饱和碳酸钾水溶液碱化到pH＝11。反应混合物用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和食盐水反洗(2×50mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得混合物未进一步纯化，直接使用。
MS(ESI,m/z):269.20[M+H]+，RT(min):0.527
第二步9-溴-苯并[4,5]咪唑并[1,2-a]哌啶(化合物21-4)的合成：
在氮气保护和冰浴下向4-(4-溴-1H-1,3-苯并二唑-2-基)丁-1-醇(2g,7.431mmol,1eq)和三苯基膦(2.92g,11.146mmol,1.5eq)的甲苯(20mL)溶液中，滴加偶氮二甲酸二叔丁酯(2.57g,11.146mmol,1.5eq)。反应液升温至85摄氏度下继续搅拌过夜。反应混合物室温下加入水(60mL)淬灭。反应混合物用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和食盐水反洗(2×80mL)，硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，二氯甲烷/甲醇(10:1)，9-溴-苯并[4,5]咪唑并[1,2-a]哌啶(化合物21-4，1g,53.59％)。
MS(ESI,m/z):250.90[M+H]+，RT(min):0.746
第三步苯并[4,5]咪唑并[1,2-a]哌啶-9-基氨基甲酸叔丁酯(化合物21-6)的合成：
氮气保护，室温下向9-溴-苯并[4,5]咪唑并[1,2-a]哌啶(500mg,1.991mmol,1eq)和氨基甲酸叔丁酯(466.49mg,3.982mmol,2eq)的1,4-二氧六环(5mL)溶液中，加入双环已基(3-异丙氧-2′,4′,6′-三异丙基-[1,1′-联苯]-2-基)膦烷(159.72mg,0.299mmol,0.15eq)，(甲磺酸{双环己基(3-异丙氧-2',4′,6′-三异丙基-[1,1′-联苯]-2-基)膦烷}(2'-甲氨基-1,1'-联苯-2-基)钯(II)(106.48mg,0.199mmol,0.1eq)和碳酸铯(1946.14mg,5.973mmol,3eq)。加入完毕后体系90摄氏度下继续搅拌4小时。反应液冷却至室温，混合物室温下加入水(30mL)稀释。反应混合物用乙酸乙酯萃取(3×30mL)。合并有机相，用饱和食盐水反洗(1×40mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(3:1)，得到苯并[4,5]咪唑并[1,2-a]哌啶-9-基氨基甲酸叔丁酯(化合物21-6，400mg,69.91％)。
MS(ESI,m/z):288.40[M+H]+，RT(min):0.728
第四步苯并[4,5]咪唑并[1,2-a]哌啶-9-胺(化合物21-7)的合成：
氮气保护，室温下向苯并[4,5]咪唑并[1,2-a]哌啶-9-基氨基甲酸叔丁酯(400mg,1.392mmol,1eq)的二氯甲烷(4mL)溶液中，加入盐酸的1,4-二氧六环溶液(4mL)。加入完毕后体系室温下继续搅拌2小时。液质检测到目标产物。所得残余物减压浓缩。所得混合物未进一步纯化，直接使用。
MS(ESI,m/z):188.05[M+H]+，RT(min):0.602
第五步4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-9-基)苯甲酰胺(化合物21-9)的合成：
氮气保护，室温下向苯并[4,5]咪唑并[1,2-a]哌啶-9-胺(240.73mg,0.641mmol,1.2eq)的二氯甲烷(2mL)溶液中加入磷酸钾(340.08mg,1.602mmol,3eq)，搅拌反应1分钟后，室温下加入4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰氯(100mg,0.534mmol,1eq)和N,N-二异丙基乙胺(207.08mg,1.602mmol,3eq)的二氯甲烷(1mL)溶液。反应液室温反应2小时。反应混合物室温下用水(20mL)淬灭。反应混合物用二氯甲烷萃取(3×20mL)。合并有机相，用饱和食盐水反洗(1×30mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。 所得混合物未进一步纯化，直接使用。
MS(ESI,m/z):526.80[M+H]+，RT(min):1.178
第六步4-((2-羟乙基)磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-9-基)苯甲酰胺(化合物21)的合成：
氮气保护，室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-9-基)苯甲酰胺(80mg,0.152mmol,1eq)的N,N-二甲基甲酰胺(2mL)溶液中加入2-羟基乙磺酰胺(28.53mg,0.228mmol,1.5eq)，碳酸钾(63.01mg,0.456mmol,3eq)，碘化亚铜(2.89mg,0.015mmol,0.1eq)和2-(甲氨基)乙酸(2.71mg,0.030mmol,0.2eq)。加入完毕后体系120摄氏度下继续搅拌2小时。反应混合物冷却到室温。加水(5mL)稀释，并用乙酸乙酯萃取(3×10mL)。合并有机相，用饱和食盐水反洗(1×10mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。粗品通过高效液相纯化，条件如下(层析柱规格:Kinetex EVO C18 Column,30*150,5um；流动相A:水(10mmol/L碳酸氢铵)，流动相B:乙腈；流速:60mL/min；洗脱梯度:10％B至50％B，8min,50％B；检测波长:UV 220nm；保留时间(分钟):7.47)，得到4-((2-羟乙基)磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-9-基)苯甲酰胺(化合物21，14.1mg,17.63％)。
MS(ESI,m/z):523.85[M+H]+，RT(min):1.157
1H NMR:(400MHz,DMSO-d6)δ11.41(s,1H),10.09(s,1H),7.87(d,1H),7.46(s,1H),7.25–7.12(m,2H),7.09–6.99(m,2H),4.97(s,1H),4.21(t,2H),3.77(t,2H),3.36–3.32(m,2H),3.04(t,4H),2.96(s,2H),1.93(d,2H),1.87(s,2H),1.52(t,4H),0.34(s,4H).
实施例3
4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)苯甲酰胺(化合物22)
第一步(Z)-N-(2,6-二溴苯基)哌啶-2-亚胺(化合物22-3)的合成：
氮气保护，0摄氏度下向2-哌啶酮(790.15mg,7.971mmol,2.0eq)的甲苯溶液中加入三氯氧磷(611.03mg,3.985mmol,1eq)，反应液0摄氏度下搅拌反应2小时后，加入2,6-二溴苯胺(1g,3.985mmol,1eq)，反应液110摄氏度下搅拌16小时。反应混合物冷却到室温。所得残余物减压浓缩，反应混合物0摄氏度下加入水(50mL)淬灭。反应混合物用氢氧化钠碱化到pH＝10。水相用二氯甲烷萃取(3×50mL)，合并有机相，用饱和食盐水反洗(1×80mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，粗品用二氯甲烷/石油醚(1:100)重结晶得到白色固体的(Z)-N-(2,6-二溴苯基)哌啶-2-亚胺(化合物22-3，900mg,68.01％)。
MS(ESI,m/z):332.75[M+H]+，RT(min):0.731
第二步6-溴-苯并[4,5]咪唑并[1,2-a]哌啶(化合物22-4)的合成：
氮气保护,室温下向(Z)-N-(2,6-二溴苯基)哌啶-2-亚胺(850mg,2.560mmol,1eq)的乙腈溶液(10mL)中加入碘化亚铜(1.43mg,0.008mmol,0.05eq),碳酸钾(20.81mg,0.151mmol,1eq)和N,N-二甲基乙二胺(1.33mg,0.015mmol,0.1eq)，反应液100摄氏度下搅拌反应2小时。反应液冷却至室温，用水稀释(30mL)，并用乙酸乙酯萃取(3×30mL)，合并有机相，用饱和食盐水反洗(2×40mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，所得残余物减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(6:1)，得到6-溴-苯并[4,5]咪唑并[1,2-a]哌啶(化合物22-4，360mg,56.00％)。
MS(ESI,m/z):250.85[M+H]+，RT(min):0.646
第三步(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)氨基甲酸叔丁酯(化合物22-5)的合成：
氮气保护,室温下向6-溴-苯并[4,5]咪唑并[1,2-a]哌啶(500mg,1.991mmol,1eq)的N,N-二甲基甲酰胺(15mL)溶液中加入碳酸铯(1946.14mg,5.973mmol,3eq)，氨基甲酸叔丁酯(466.49mg,3.982mmol,2eq)，(甲磺酸{双环己基(3-异丙氧-2',4′,6′-三异丙基-[1,1′-联苯]-2-基)膦烷}(2'-甲氨基-1,1'-联苯-2-基)钯(II)(182.78mg,0.199mmol,0.1eq)和二环己基(3-乙丙氧基-2′,4′,6′-三异丙基-[1,1′-二苯基]-2-基)膦；二环己基(3-异丙基-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦(212.96mg,0.398mmol,0.2eq)，反应液100摄氏度搅拌反应1小时。反应液冷却至室温，用水稀释(20mL)，反应混合物用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水反洗(1×20mL)，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(2:1)，得到(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)氨基甲酸叔丁酯(化合物22-5，490mg,85.64％)。
MS(ESI,m/z):288.35[M+H]+,RT(min):0.768
第四步苯并[4,5]咪唑并[1,2-a]哌啶-6-胺(化合物22-6)的合成：
氮气保护，室温下向(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)氨基甲酸叔丁酯(400mg,1.392mmol,1eq)的二氯甲烷(5mL)溶液中加入氯化氢(气体)溶于1,4-二氧六环(5mL),搅拌反应1小时。所得残余物减压浓缩,得到苯并[4,5]咪唑并[1,2-a]哌啶-6-胺粗品，未进一步纯化，直接投下步。
MS(ESI,m/z):188.35[M+H]+，RT(min):0.450
第五步4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)苯甲酰胺(化合物22-7)的合成：
氮气保护,室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酸(629.51mg,1.763mmol,1.5eq)的二氯甲烷溶液(8mL)中加入氯化亚砜(279.54mg,2.350mmol,2eq)，反应液室温搅拌反应1小时。所得残余物减压浓缩。所得残余物用二氯甲烷溶解(4mL)，室温下向上述体系中加入磷酸钾(498.79mg,2.350mmol,2eq),室温搅拌5分钟，然后向上述体系中加入苯并[4,5]咪唑并[1,2-a]哌啶-6-胺(20mg,0.107mmol,1eq)和N,N-二异丙基乙胺(455.57mg,3.525mmol,3eq)的二氯甲烷溶液(4mL)，加入完毕后体系室温下继续搅拌2小时。反应液加水淬灭(20mL)，反应混合物用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水反洗(1×20mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)苯甲酰胺(化合物22-7，530mg,85.69％)。
MS(ESI,m/z):527.35[M+H]+，RT(min):1.078
第六步4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)苯甲酰胺(化合物22)的合成：
氮气保护,室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)苯甲酰胺(100mg,0.190mmol,1eq)的N,N-二甲基甲酰胺溶液(2mL)中加入碘化亚铜(3.62mg,0.019mmol,0.1eq)，肌氨酸(3.39mg,0.038mmol,0.2eq)，碳酸钾(78.76mg,0.570mmol,3eq)和2-羟基乙磺酰胺(47.54mg,0.380mmol,2eq)，反应液120摄氏度下搅拌反应1.5小时。反应液冷却至室温，并用水稀释(10mL)，反应混合物用乙酸乙酯萃取(3×10mL)。合并有机相，用饱和食盐水反洗(1×10mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。粗品通过高效液相纯化，条件如下(层析柱规格:Xselect CSH C18 OBD Column 30*150mm 5μm,n；流动相A:水(0.1％甲酸),流动相B:乙腈；流速:60mL/min；洗脱梯度:10％B至37％B，10min,37％B；检测波长:220nm；保留时间(分钟):9.03；运行次数:0)。得到4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(苯并[4,5]咪唑并[1,2-a]哌啶-6-基)苯甲酰胺(化合物22，30.7mg,27.69％)。
MS(ESI,m/z):523.90[M+H]+，RT(min):1.530
1H NMR(400MHz,DMSO-d6)δ12.44(s,1H),10.14(s,1H),8.33(d,1H),8.04(d,1H),7.27(d,1H),7.21–7.13(m,2H),7.09(dd,1H),4.93(d,1H),4.13(t,2H),3.76(q,2H),3.32(s,2H),3.00(q,6H),2.11–2.03(m,2H),2.02–1.92(m,2H),1.83(s,4H),0.31(s,4H).
实施例4
N-(2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物36)
第一步(2Z)-N-(2,6-二溴苯基)吡咯烷-2-亚胺(化合物36-2)的合成：
氮气保护，0℃下向吡咯烷酮(1.36g,15.942mmol,2eq)的甲苯(10mL)溶液中，加入三氯氧磷(1.22g,85.106mmol,2eq)，反应液0℃反应2小时，随后加入2,6-二溴苯胺(2g,7.971mmol,1eq)，反应液升温至110℃，搅拌反应16小时，反应液冷却至室温，反应液减压浓缩，加冰水(50mL)淬灭，用饱和碳酸氢钠溶液将PH值调至10，并用乙酸乙酯萃取(3×50mL)，合并有机相，用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用制备色谱板纯化，乙酸乙酯/石油醚(1:10)，得到(2Z)-N-(2,6-二溴苯基)吡咯烷-2-亚胺(2.7g,27.65％)。
MS(ESI,m/z):317.10[M+H]+,RT(min):0.919
第二步5-溴-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑(化合物36-3)的合成：
氮气保护，室温下向(2Z)-N-(2,6-二溴苯基)吡咯烷-2-亚胺(2.7g,8.490mmol,1eq)的乙腈(3mL)溶液中加入N,N'-二甲基-1,2-乙二胺(74.84mg,0.849mmol,0.10eq)，碘化亚铜(80.85mg,0.425mmol,0.05eq)，碳酸钾(1.17g,8.490mmol,1eq)，反应液升温至100℃，搅拌2小时。反应液冷却至室温，加水(80mL)淬灭，并用乙酸乙酯萃取(3×60mL)。合并有机相，用饱和食盐水反洗(1×80mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。得到5-溴-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑(2g,74.07％)。
MS(ESI,m/z):237.15[M+H]+,RT(min):0.657
第三步(2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)氨基甲酸叔丁酯(化合物36-5)的合成：
氮气保护，室温下向5-溴-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑(2g,8.436mmol,1eq)，氨基甲酸叔丁酯(1.99g,16.87mmol,2eq)的N,N-二甲基甲酰胺(10mL)溶液中加入二环己基(3-异丙基-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦(451.12mg,0.844mmol,0.1eq),(甲磺酸{双环乙基(3-异丙基-2,4,6-三异丙基-(1,1-联苯)-2-基)膦烷})(2-甲氨基-1,1-联苯-2-基)钯(Ⅱ)(1.549g,1.688mmol,0.2eq)和碳酸铯(8.245g,25.308mmol,3eq)。反应液升温至100℃，反应液搅拌反应1小时。反应液冷却至室温，加水(80mL)淬灭。并用乙酸乙酯萃取(3×60mL)。合并有机相，用饱和食盐水反洗(1×80mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用制备色谱板纯化，乙酸乙酯/石油醚(1:5)，得到(2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)氨基甲酸叔丁酯(700mg,35.77％)。
MS(ESI,m/z):274.35[M+H]+,RT(min):0.592
第四步2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-胺(化合物36-6)的合成：
(2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)氨基甲酸叔丁酯(300mg,1.098mmol,1eq)，盐酸的1,4-二氧六环(2mL)，二氯甲醇(2mL)的反应液在室温下搅拌30分钟，反应液减压浓缩，得到2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-胺(300mg,57.87％)，直接投下一步。
MS(ESI,m/z):174.35[M+H]+,RT(min):0.602
第五步2-{6-氮杂螺[2.5]辛烷-6-基}-N-{2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基}-4-碘苯甲酰胺(化合物36-7)的合成：
氮气保护，室温下向2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-胺(250mg,1.443mmol,1eq)、2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(618.62mg,1.732mmol,1.2eq)的二氯甲烷(5mL)溶液中加入四甲基氯代脲六氟磷酸酯(1.62g,5.772mmol,4eq)，N-甲基咪唑(1.18g,14.430mmol,10eq)，反应液室温搅拌反应2小时。反应液加水(20mL)稀释，并用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水反洗(1×20mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用制备色谱板纯化(乙酸乙酯:石油醚＝1:5)，得到2-{6-氮杂螺[2.5]辛烷-6-基}-N-{2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基}-4-碘苯甲酰胺.(100mg,40.00％)。
MS(ESI,m/z):513.35[M+H]+,RT(min):1.406
第六步N-(2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物36)的合成：
氮气保护，室温下向N-(2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(60mg,0.117mmol,1eq)和2-羟基乙磺酰胺(17.58mg,0.140mmol,1.2eq)的N,N-二甲基甲酰胺(2mL)溶液中加入肌氨酸(2.09mg,0.023mmol,0.2eq)，碘化亚铜(2.23mg,0.012mmol,0.1eq)和碳酸钾(48.55mg,0.351mmol,3eq)，反应液升温至120℃，反应液搅拌反应1.5小时。反应液冷却至室温，加水淬灭，反应混合物用乙酸乙酯萃取(3×10mL)。合并有机相，用饱和食盐水溶液反洗(1×10mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得粗品用制备型高压液相纯化，条件如下:层析柱规格:Kinetex EVO C18色谱柱,30*150,5μm；流动相A:水(10mmol/L碳酸氢铵)，流动相B:乙腈；流速:60mL/min；洗脱梯度:30％B to 65％B in 8min,60％B；检测波长:UV 220nm；保留时间(分钟):6.12，得到N-(2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(4.22mg,6.60％)。
MS(ESI,m/z):510.15[M+H]+,RT(min):1.143
1H NMR:(400MHz,DMSO-d6)δ12.34(s,1H),10.14(s,1H),8.30(dd,1H),8.01(d,1H),7.25(d,1H),7.19–7.10(m,2H),7.09–7.04(m,1H),4.96(s,1H),4.15(t,2H),3.76(t,2H),3.36(d,2H),2.97(dt,6H),2.67(q,2H),1.78(s,4H),0.30(s,4H).
使用与实施例4中类似的条件，制备了如下表中的化合物。

实施例5
2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}-4-(2-羟基乙磺酰氨基)苯甲酰胺(化合物51)
第一步5'-溴-3',4'-二氢螺[环戊烷-1,2'-萘]-1'-酮(化合物51-3)的合成：
氮气保护，0℃下向5-溴-3,4-二氢-2H-萘-1-酮(2g,8.886mmol,1eq)的四氢呋喃(20mL)溶液中加入氢化钠(0.71g,29.586mmol,3.33eq)，继续搅拌1小时后，0℃下滴加1,4-二碘代丁烷(4.13g,13.329mmol,1.5e)。反应液升温至室温，搅拌反应16小时。反应混合物室温下用冰水淬灭。反应混合物用乙酸乙酯萃取(3×80mL)。合并有机相，用饱和食盐水溶液反洗(1×100mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(10:1)，得到5'-溴-3',4'-二氢螺[环戊烷-1,2'-萘]-1'-酮(2.37g,80.56％)。
MS:(ESI,m/z):278.55[M+H]+,RT(min):1.325
第二步5'-溴-3',4'-二氢-1'H-螺[环戊烷-1,2'-萘](化合物51-4)的合成：
氮气保护，0℃下向5'-溴-3',4'-二氢螺[环戊烷-1,2'-萘]-1'-酮(2g,7.11mmol,1eq)的三氟乙酸(15mL)溶液中加入三乙基硅烷(5.0mL,30.94mmol,4.35eq)，反应液升温至50℃，搅拌反应1小时。反应液冷却至室温，反应混合物用乙酸乙酯萃取(3×80mL)。合并有机相，用饱和食盐水溶液反洗(1×80mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(10:1)，得到5'-溴-3',4'-二氢-1'H-螺[环戊烷-1,2'-萘](1.43g,64.18％).
MS(EI,m/z):264.1[M],RT(min):8.694
第三步{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}氨基甲酸叔丁酯(化合物51-5)的合成：
氮气保护，室温下向5'-溴-3',4'-二氢-1'H-螺[环戊烷-1,2'-萘](1g,3.75mmol,1eq)，氨基甲酸叔丁酯(883.5mg,7.5mmol,2eq)的1,4-二氧六环(10mL)溶液中加入碳酸铯(3.7g,12.75mmol,3eq)，醋酸钯(84.5mg,0.35mmol,0.1eq)和2-双环己基膦-2,4,6-三异丙基联苯(359.5mg,0.75mmol,0.2eq)。反应液升温至100℃，搅拌反应1小时。反应液冷却至室温，并用水淬灭，反应混合物用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和食盐水溶液反洗(1×80mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(10:1)，得到{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}氨基甲酸叔丁酯(800mg,70.79％)。
MS(EI,m/z):201.1[M-100],RT(min):9.866
第四步3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-胺(化合物51-6)的合成：
室温下向{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}氨基甲酸叔丁酯(200mg,0.66mmol,1eq)的二氯甲烷溶液(2mL)中加入盐酸的1,4-二氧六环溶液(2mL)，反应液室温搅拌反应30分钟。反应液减压浓缩，得到3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-胺(60mg,46.78％)。粗品未进一步纯化，直接投下步。
MS:(ESI,m/z):202.45[M+H]+,RT(min):1.041
第五步2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}-4-碘苯甲酰胺(化合物51-7)的合成：
氮气保护，室温下向2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(127.74mg,0.36mmol,1.2eq)的二氯甲烷(1mL)溶液中加入N,N,N,N-四甲基氯甲脒六氟磷酸盐(334.5mg,1.12mmol,4eq)，N-甲基咪唑(244.71mg,2.97mmol,10eq)和3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-胺(60mg,0.300mmol,1eq)。反应液室温下搅拌反应1小时。反应混合物用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水溶液反洗(1×20mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(10:1)，得到2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}-4-碘苯甲酰胺(100mg,90.34％)。
MS(ESI,m/z):541.35[M+H]+,RT(min):1.728
第六步2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}-4-(2-羟基乙磺酰氨基)苯甲酰胺(化合物51)的合成：
氮气保护，室温下向2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}-4-碘苯甲酰胺(50mg,0.095mmol,1eq)，2-羟基乙磺酰胺(13.9mg,0.115mmol,1.2eq)的N,N-二甲基甲酰胺(1mL)溶液中加入碘化亚铜(1.75mg,0.010mmol,0.1eq)，肌氨酸(1.65mg,0.020mmol,0.20eq)，碳酸钾(38.35mg,0.285mmol,3eq)。反应液升温至120℃，搅拌反应1小时。反应液冷却至室温，加水淬灭，反应混合物用乙酸乙酯萃取(3×10mL)。合并有机相，用饱和食盐水溶液反洗(1×10mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得粗品用制备型高压液相纯化，条件如下:层析柱规格:Kinetex EVO C18色谱柱,30*150,5μm；流动相A:水(10mmol/L碳酸氢铵)，流动相B:乙腈；流速:60mL/min；洗脱梯度:45％B to 96％B in 8min,96％B；检测波长:UV 220nm；保留时间(分钟):7.62，得到2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环戊烷-1,2'-萘]-5'-基}-4-(2-羟基乙磺酰氨基)苯甲酰胺(10mg,20.06％)。
MS(ESI,m/z):538.10[M+H]+,RT(min):1.383
1H NMR:(400MHz,DMSO-d6)δ10.87(s,1H),10.07(s,1H),7.85(d,1H),7.58(m,1H),7.19(d,1H),7.17(d,1H),7.03(m,2H),4.93(d,1H),3.76(q,2H),2.99(d,4H),2.78(d,2H),2.07(s,2H),1.89(s,1H),1.78(s,1H),1.70(d,6H),1.46–1.36(m,4H),1.30–1.13(m,4H),0.34(s,4H).
实施例6
2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-4-(2-羟基乙磺酰氨基)苯甲酰胺(化合物56)
第一步碘化(2-氯乙基)二甲基硫鎓(化合物56-2)的合成：
1-氯-2-(甲硫基)乙烷(8g,72.333mmol,1eq)和碘甲烷(27.02mL,433.998mmol,6eq)的溶液在室温下搅拌65小时。混合物中有固体析出，过滤，滤饼用丙酮(3×30mL)洗涤，固体干燥，得到碘化(2-氯乙基)二甲基硫鎓(9.5g,52.01％)。
MS(ESI,m/z):125.02[M]+.RT(min):0.19.
第二步5'-溴-3',4'-二氢螺[环丙烷-1,2'-萘]-1'-酮(化合物56-4)的合成：
在0℃下，向5-溴-3,4-二氢-2H-萘-1-酮(2g,10.461mmol,1.00eq)的叔丁醇(30mL)溶液中，加入碘化钠(0.27g,1.777mmol,0.2eq)，氢化钠(0.71g,60％,分散在煤油中，17.77mmol,2eq)。混合溶液室温下搅拌20分钟，然后分批加入碘化(2-氯乙基)二甲基硫鎓(2.47g,9.775mmol,1.1eq)。搅拌反应16小时。反应溶液用水淬灭，用乙酸乙酯(3×50mL)萃取。合并有机相，用饱和食盐水反洗(2×50mL)，所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到5'-溴-3',4'-二氢螺[环丙烷-1,2'-萘]-1'-酮(1.8g,80.67％)。
MS(EI,m/z):250.1[M]+.RT(min):8.46.
第三步5'-溴-3',4'-二氢-1'H-螺[环丙烷-1,2'-萘](化合物56-5)的合成：
氮气保护下，0℃下向5'-溴-3',4'-二氢螺[环丙烷-1,2'-萘]-1'-酮(800mg,3.186mmol,1eq)的二氯甲烷(30mL)溶液中加入三乙基硅烷(2.57mL,15.930mmol,5eq)，然后滴加三氟化硼乙醚溶液(2.02mL,15.930mmol,5eq)。反应液0℃下搅拌4小时，随后升至室温，搅拌16小时。反应溶液用饱和的碳酸氢钠溶液淬灭，用二氯甲烷(3×50mL)萃取。合并有机相，用饱和食盐水反洗(1×60mL)，无水硫酸钠干燥，过滤， 滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(10:1)，得到5'-溴-3',4'-二氢-1'H-螺[环丙烷-1,2'-萘](550mg,粗品)。
第四步N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-1,1-二苯基甲亚胺(化合物56-7)的合成：
氮气保护下，室温下向5'-溴-3',4'-二氢-1'H-螺[环丙烷-1,2'-萘](500mg,2.108mmol,1eq)，二苯甲酮亚胺(458.56mg,2.530mmol,1.2eq)，叔丁醇钠(303.95mg,3.162mmol,1.5eq)的甲苯(20mL)的溶液中，加入1,1'-联萘-2,2'-双二苯膦(131.29mg,0.211mmol,0.1eq)和三(二亚苄基丙酮)二钯(96.54mg,0.105mmol,0.05eq)。反应液升温至90℃，搅拌反应16小时。反应液冷却至室温，过滤，滤饼用乙酸乙酯洗涤，混合物用乙酸乙酯萃取(3×50mL)，合并有机相，用饱和食盐水反洗(1×60mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，所得粗品N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-1,1-二苯基甲亚胺(1.1g,粗品)未进一步纯化，直接投下步。
MS(ESI,m/z):338.50[M+H]+.RT(min):1.524.
第五步3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-胺(化合物56-8)的合成：
N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-1,1-二苯基甲亚胺(1.1g,粗品)的1M的盐酸(10mL)室温搅拌16小时，反应液用饱和碳酸氢钠水溶液碱化到pH＝8。反应混合物用二氯甲烷萃取(3×50mL)。合并有机相，用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，二氯甲烷/甲醇(20:1)，得到3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-胺(100mg)。
MS(ESI,m/z):174.45[M+H]+.RT(min):0.892.
第六步2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-4-碘苯甲酰胺(化合物56-9)的合成：
氮气保护，室温下向3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-胺(100mg,0.577mmol,1eq)，2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(247.39mg,0.692mmol,1.2eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(647.77mg,2.308mmol,4eq)的二氯甲烷(4mL)溶液中，加入N-甲基咪唑(473.89mg,5.770mmol,10eq)。反应液升温至80℃，反应2小时。反应液冷却至室温，用二氯甲烷萃取(3×20mL)，合并有机相，用饱和食盐水反洗(1×30mL)，无水硫酸干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(3:1)，得到2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-4-碘苯甲酰胺(56mg,18.93％)。
MS(ESI,m/z):513.05[M+H]+.RT(min):1.262.
第七步2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-4-(2-羟基乙磺酰氨基)苯甲酰胺(化合物56)的合成：
氮气保护，室温下向2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-4-碘苯甲酰胺(50mg,0.098mmol,1eq)，2-羟基乙磺酰胺(14.65mg,0.118mmol,1.2eq)和碳酸钾(40.46mg,0.294mmol,3eq)的N,N-二甲基甲酰胺(1mL)溶液中，加入碘化亚铜(1.86mg,0.010mmol,0.1eq)和肌氨酸(1.74mg,0.020mmol,0.2eq)。反应液升温至120℃，反应1小时。反应液冷却至室温，用乙酸乙酯萃取(3×20mL)，合并有机相，用饱和食盐水反洗(3×10mL)，无水硫酸干燥，过滤，滤液减压浓缩，粗品通过制备型高效液相纯化，条件如下(层析柱规格：Xselect CSH C18 OBD色谱柱，30*150mm 5μm；流动相A:水(0.1％甲酸)，流动相B:乙腈；流速:60mL/min；洗脱梯度：45％B to 75％B in 8min；检测波长:UV 220nm；保留时间(分钟):7.4)，得到2-{6-氮杂螺[2.5]辛烷-6-基}-N-{3',4'-二氢-1'H-螺[环丙烷-1,2'-萘]-5'-基}-4-(2-羟基乙磺酰氨基)苯甲酰胺(12mg,24.01％).
MS(ESI,m/z):510.55[M+H]+.RT(min):1.926.
1H NMR(400MHz,DMSO-d6)δ10.93(s,1H),10.08(s,1H),7.86(d,1H),7.65(d,1H),7.18(d,1H),7.12(t,1H),7.02(dd,1H),6.88(d,1H),4.95(s,1H),3.76(t,2H),3.36–3.30(m,2H),3.00(t,4H),2.79(t,2H),2.66(s,2H),1.57(t,2H),1.47(s,4H),0.39(s,4H),0.33(s,4H).
实施例7
N-(8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物64)

第一步5,5-二氟-1-(4-甲基-3-硝基吡啶-2-基)哌啶-2-酮(化合物64-3)的合成：
氮气保护下，室温下向2-氯-4-甲基-3-硝基吡啶(2.4g,13.907mmol,1eq)，5,5-二氟哌啶-2-酮(2.25g,16.688mmol,1.2eq)的1,4-二氧六环(50mL)溶液中加入醋酸钯(156.12mg,0.695mmol,0.05eq)，4,5-双二苯基膦-9,9-二甲基氧杂蒽(804.73mg,1.391mmol,0.1eq)和碳酸铯(6.80g,20.861mmol,1.5eq)。反应液升温至100摄氏度，搅拌16小时。反应液冷却至室温，减压浓缩，所得残余物用硅胶柱层析纯化，正己烷/乙酸乙酯(6:1)，得到5,5-二氟-1-(4-甲基-3-硝基吡啶-2-基)哌啶-2-酮(化合物64-3，980mg,25.98％)。
MS(ESI,m/z):272.30[M+H]+.RT(min):0.786
第二步8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶(化合物64-4)的合成：
室温下向5,5-二氟-1-(4-甲基-3-硝基吡啶-2-基)哌啶-2-酮(1.06g,3.908mmol,1eq)的乙醇和水溶液(10mL:1mL)中加入铁粉(12.35mg,0.222mmol,3eq)和氯化铵(0.13g,2.345mmol,0.6eq)。反应液升温至80摄氏度，搅拌16小时。反应液冷却至室温，过滤，滤液浓缩至干，所得残余物用硅胶柱纯化，石油醚/乙酸乙酯(4:1)，得到8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶(化合物64-4,550mg,63.04％)。
MS(ESI,m/z):224.30[M+H]+.RT(min):0.666
第三步8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-1-氧化物(化合物64-5)的合成：
室温下向8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶(500mg,1.843mmol,1eq)的二氯甲烷溶液(5mL)中，加入间氯过氧苯甲酸(561.38mg,2.764mmol,1.5eq,85％)，在室温下搅拌16小时，反应液直接减压浓缩，所得残余物用硅胶柱柱层析，石油醚/乙酸乙酯(1:3)，得到8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-1-氧化物(化合物64-5，220mg,53.46％)。
MS(ESI,m/z):240.30[M+H]+.RT(min):0.585
第四步1-(8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)吡啶-1-三氟乙酸盐(化合物64-6)的合成：
室温下向8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-1-氧化物(220mg,0.920mmol,1eq)的二氯甲烷溶液(4mL)中加入吡啶(0.37mL,4.600mmol,5eq)，在0摄氏度，滴加三氟乙酸酐(0.38mL,2.760mmol,3eq)，反应液升温至室温搅拌16小时。所得混合物未进一步纯化，直接使用。
MS(ESI,m/z):301.30[M-CF3COO-]+.RT(min):0.501
第五步8,8-二氟-4-甲基吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-胺(化合物64-7)的合成：
室温下向1-(8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)吡啶-1-三氟乙酸盐(220mg,0.53mmol,1eq)的乙醇溶液(3mL)中加入乙醇胺(0.32mL,5.310mmol,10eq)，反应液升温至80摄氏度，搅拌16小时，反应液减压浓缩，所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(2:1)，得到8,8-二氟-4-甲基吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-胺(化合物64-7，75mg)。
MS(ESI,m/z):239.35[M+H]+.RT(min):0.475.
第六步N-(8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物64-8)的合成：
氮气保护，室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酸(112.45mg,0.315mmol,1.00eq)，8,8-二氟-4-甲基吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-胺(75mg,0.315mmol,1.00eq)，四甲基氯甲脒六氟磷酸盐(353.31mg,1.260mmol,4eq)的二氯甲烷(4mL)的溶液中加入N-甲基咪唑(0.25mL,3.150mmol,10eq)。反应液升温至60摄氏度反应2小时。反应液冷却至室温，用水稀释(10mL)，并用乙酸乙酯萃取(3×10mL)，合并有机相，用饱和食盐水反洗(2×10mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，所得残余物用制备色谱板纯化，石油醚/乙酸乙酯(6:1)，得到N-(8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物64-8，30mg,16.50％)。
MS(ESI,m/z):578.30[M+H]+.RT(min):1.432
第七步N-(8,8-二氟-4-甲基-吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物64)的合成：
氮气保护下，室温下向N-(8,8-二氟-4-甲基吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(30mg,0.052mmol,1eq)，2-羟基乙基磺酰胺(7.80mg,0.062mmol,1.2eq)，肌氨酸(0.93mg,0.010mmol,0.2eq)和碳酸钾(21.54mg,0.156mmol,3eq)的二甲基甲酰胺(0.8mL)溶液加入碘化亚铜(0.99mg,0.005mmol,0.1eq)。反应液升温至120摄氏度反应1小时。反应液冷却至室温，用水稀释(10mL)，并用乙酸乙酯萃取(3×10mL)，合并有机相，用饱和食盐水反洗(2×10mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，所得残余物通过高效液相纯化，条件如下(层析柱规格:Kinetex EVO prep C18,30*150,5μm；流动相A:水(10mmol/L碳酸氢铵),流动相B:乙腈；流速:60mL/min；洗脱梯度:20％B至55％B，8min,55％B；检测波长:220nm；保留时间(分钟):7.57)。得到N-(8,8-二氟-4-甲基吡啶并[2,3-d]咪唑并[1,2-a]哌啶-2-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物64，2.24mg,7.46％)。
MS(ESI,m/z):575.35[M+H]+.RT(min):2.150
1H NMR(400MHz,DMSO-d6)δ13.57(s,1H),8.11–8.02(m,2H),7.25(s,1H),7.11(d,1H),4.47(t,2H),3.76(m,4H),3.33(m,2H),3.19(m,2H),2.99(t,4H),2.61(m,2H),2.56(s,3H),1.81(s,4H),0.40(s,4H).
实施例8
N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物94)
第一步(2Z)-N-(2,6-二溴苯基)-3,3-二氟吡咯烷-2-亚胺(化合物94-2)的合成：
氮气保护，0℃下向3,3-二氟吡咯烷-2-酮(0.99g,8.176mmol,1.50eq)的超干甲苯(15mL)溶液中加入POCl3(837.11mg,5.460mmol,1eq)，反应液室温搅拌2小时。向上述溶液中加入2,6-二溴苯胺(1.37g,5.460 mmol,1eq)的甲苯(5mL)溶液，反应液升温至110℃，搅拌16小时。反应液冷却到室温，加水淬灭。反应混合物用二氯甲烷萃取(3×60mL)。合并有机相，用饱和氯化钠溶液反洗(1×80mL),无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用石油醚/二氯甲烷(100:1)重结晶得到(2Z)-N-(2,6-二溴苯基)-3,3-二氟吡咯烷-2-亚胺(1.2g,62.09％)。
MS(ESI,m/z):352.85[M+H]+,RT(min):0.674
第二步5-溴-3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑(化合物94-3)的合成：
氮气保护，室温下向(2Z)-N-(2,6-二溴苯基)-3,3-二氟吡咯烷-2-亚胺(500mg,1.412mmol,1eq)的N,N-二甲基甲酰胺(10.00mL)中加入N,N-二甲基乙二胺(12.45mg,0.141mmol,0.1eq)，无水碳酸钾(195.21mg,1.412mmol,1eq)和碘化亚铜(13.45mg,0.071mmol,0.05eq)。反应液升温至100℃,搅拌反应1h。反应液冷却至室温，加水淬灭，反应混合物用二氯甲烷萃取(3×50mL)。合并有机相，用饱和食盐水溶液反洗(1×50mL),无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(2:1)，得到5-溴-3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑(240mg,62.22％)。
MS(ESI,m/z):272.90[M+H]+,RT(min):0.837
第三步(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)氨基甲酸叔丁酯(化合物94-4)的合成：
氮气保护，室温下向5-溴-3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑(230mg,0.842mmol,1eq)的N,N-二甲基甲酰胺(5.00mL)溶液中加入氨基甲酸叔丁酯(197.33mg,1.684mmol,2eq)，(甲磺酸{双环己基(3-异丙氧-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦烷}(2'-甲氨基-1,1'-联苯-2-基)钯(II)(154.73mg,0.168mmol,0.20eq),二环己基(3-异丙基-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦(45.04mg,0.084mmol,0.10eq),碳酸铯(823.25mg,2.526mmol,3.00eq)，反应液升温至100℃，搅拌反应1h。液质监测反应完全。反应混合物用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和氯化钠溶液反洗(1×50mL),无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(2:1)，得到(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)氨基甲酸叔丁酯(189mg,72.55％)。
MS(ESI,m/z):310.05[M+H]+,RT(min):1.004
第四步3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-胺(化合物94-5)的合成：
氮气保护，室温下向(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)氨基甲酸叔丁酯(189mg,0.611mmol,1eq)的二氯甲烷溶液(2mL)中加入盐酸的1,4-二氧六环(2mL)溶液，反应液室温搅拌反应1小时。反应液减压浓缩。得到3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-胺(127mg,99.35％)。
MS(ESI,m/z):210.10[M+H]+,RT(min):0.193
第五步N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物94-6)的合成：
氮气保护，室温下向2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(204.89mg,0.574mmol,1.2eq)的二氯甲烷溶液(10.00mL)中加入3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-胺(100mg,0.478mmol,1eq)，四甲基氯甲脒六氟磷酸盐(536.48mg,1.912mmol,4eq)，N-甲基咪唑(392.48mg,4.780mmol,10eq)，反应液升温至50℃，搅拌反应1小时。反应液冷却至室温，反应液用水稀释，反应混合物用二氯甲烷萃取(3×50mL)，合并有机相，用饱和食盐水反洗(1×50mL),无水硫酸钠干燥,过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(4:1)，得到N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(150mg,57.22％)。
MS(ESI,m/z):549.05[M+H]+,RT(min):1.311
第六步N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物94)的合成：
氮气保护，室温下向N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(100mg,0.182mmol,1eq)的N,N-二甲基甲酰胺(20.00mL)溶液中加入2-羟基乙磺酰胺(27.38mg,0.218mmol,1.2eq),肌氨酸(3.25mg,0.036mmol,0.2eq)，碘化亚铜(3.47mg,0.018mmol,0.1eq),碳酸钾(75.61mg,0.55mmol,3eq)。反应液升温至140℃，搅拌反应1小时。反应液冷却至室温，加水淬灭，反应混合物用乙酸乙酯萃取(3×30mL)。合并有机相，用饱和食盐水溶液反洗(1×30mL),无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用制备型高效液相纯化，条件如下(层析柱规格:Kinetex EVO prep C18,30*150,5μm；流动相A:水(10mmol/L碳酸氢钠),流动相B:乙腈；流速:60mL/min；洗脱梯度:20％B to 65％B in 8min,65％B；检测波长:220nm；保留时间(min):7.45)。得到N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-(2-羟乙基磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(23.00mg,23.12％)。
MS(ESI,m/z):546.30[M+H]+,RT(min):1.621
1H NMR(400MHz,DMSO-d6)δ12.55(s,1H),10.16(s,1H),8.47(m,1H),8.04(d,1H),7.45–7.33(m,2H),7.28(d,1H),7.09(m,1H),4.92(s,1H),4.66–4.34(m,2H),3.76(t,2H),3.43–3.32(m,2H),3.01(d,4H),1.77(s,4H), 1.23(s,2H),0.27(s,4H).
使用与实施例8中类似的条件，制备了如下表中的化合物。
实施例9
4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(化合物96)
第一步(3Z)-N-(2,6-二溴苯基)-2-氮杂双环[2.2.1]庚烷-3-亚胺(化合物96-2)的合成：
氮气保护，0℃下向2-氮杂双环[2.2.1]庚烷-3-酮(885.89mg,7.970mmol,2eq)的无水甲苯(15mL)溶液中加入三氯氧磷(1833.09mg,11.955mmol,3eq)，反应液室温搅拌2小时。向上述溶液中加入2,6-二溴苯胺 (1g,3.985mmol,1eq)的甲苯(5mL)溶液，反应液升温至100℃，搅拌16小时。反应液冷却到室温，加水(30mL)淬灭。反应混合物用乙酸乙酯(3×80mL)萃取。合并有机相，用饱和氯化钠溶液(1×80mL)反洗，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(3:1)，得到(3Z)-N-(2,6-二溴苯基)-2-氮杂双环[2.2.1]庚烷-3-亚胺(190mg,13.86％)。
MS(ESI,m/z):344.80[M+H]+,RT(min):0.592.
第二步6-溴-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶(化合物96-3)的合成：
氮气保护，室温下向(3Z)-N-(2,6-二溴苯基)-2-氮杂双环[2.2.1]庚烷-3-亚胺(190mg,0.553mmol,1eq)的乙腈(5.0mL)溶液中加入N,N-二甲基乙二胺(4.87mg,0.055mmol,0.1eq)，无水碳酸钾(76.22mg,0.553mmol,1eq)和碘化亚铜(5.26mg,0.028mmol,0.05eq)。反应液升温至100℃，搅拌反应1小时。反应液冷却至室温，加水淬灭，反应混合物用乙酸乙酯(3×50mL)萃取。合并有机相，用饱和食盐水溶液(1×50mL)反洗，无水硫酸钠干燥。过滤，滤液减压浓缩。得到6-溴-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶(140mg,96.55％)。
MS(ESI,m/z):263.00[M+H]+,RT(min):0.602.
第三步(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(化合物96-4)的合成：
氮气保护，室温下向6-溴-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶(130mg,0.494mmol,1eq)的N,N-二甲基甲酰胺(5.0mL)溶液中加入氨基甲酸叔丁酯(115.75mg,0.988mmol,2eq)，(甲磺酸{双环己基(3-异丙氧-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦烷}(2'-甲氨基-1,1'-联苯-2-基)钯(II)(45.38mg,0.049mmol,0.1eq)，二环己基(3-异丙基-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦(52.84mg,0.099mmol,0.2eq)，碳酸铯(482.9mg,1.482mmol,3eq)，反应液升温至100℃，搅拌反应1小时。反应液冷却至室温，用乙酸乙酯(3×50mL)萃取。合并有机相，用饱和氯化钠溶液(1×50mL)反洗，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:4)，得到(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(94mg,63.56％)。
MS(ESI,m/z):300.10[M+H]+,RT(min):0.730.
第四步1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(化合物96-5)的合成：
室温下向(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(94mg,0.314mmol,1eq)的二氯甲烷(2mL)溶液中加入盐酸的1,4-二氧六环溶液(2mL)溶液，反应液室温搅拌反应1小时。反应液减压浓缩。得到1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(62mg,99.10％)。
MS(ESI,m/z):200.00[M+H]+,RT(min):0.166
第五步4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(化合物96-6)的合成：
氮气保护，室温下向1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(60mg,0.301mmol,1eq)，2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(129.07mg,0.361mmol,1.20eq)的二氯甲烷溶液(10.0mL)中加入四甲基氯甲脒六氟磷酸盐(337.95mg,1.204mmol,4eq)，N-甲基咪唑(247.24mg,3.010mmol,10eq)，反应液室温搅拌反应1小时。反应液用二氯甲烷萃取(3×50mL)，合并有机相，用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:2)，得到4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(70mg,43.17％)。
MS(ESI,m/z):539.35[M+H]+,RT(min):1.495.
第六步4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(化合物96)的合成：
氮气保护，室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(65mg,0.121mmol,1eq)，2-羟基乙磺酰胺(30.21mg,0.242mmol,2.0eq)和碳酸钾(50.05mg,0.363mmol,3eq)的N,N-二甲基甲酰胺(5.0mL)溶液中加入肌氨酸(2.16mg,0.024mmol,0.2eq)，碘化亚铜(2.3mg,0.012mmol,0.1eq)。反应液升温至120℃，搅拌反应1小时。反应液冷却至室温，加水淬灭，反应混合物用乙酸乙酯萃取(3×30mL)。合并有机相，用饱和食盐水溶液反洗(1×30mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用制备型高效液相纯化，条件如下(层析柱规格:Kinetex EVO prep C18,30*150,5μm；流动相A:水(10mmol/L碳酸氢钠)，流动相B:乙腈；流速:60mL/min；洗脱梯度:20％B to 65％B in 8min,65％B；检测波长:UV 220nm；保留时间(min):7.45)。得到4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(16.99mg,25.96％)。
MS(ESI,m/z):536.00[M+H]+,RT(min):1.488.
1H NMR(400MHz,DMSO-d6)δ12.32(s,1H),10.13(s,1H),8.28(d,1H),8.00(d,J＝8.6Hz,1H),7.31– 7.18(m,2H),7.12(t,1H),7.06(m,1H),5.15(s,1H),4.94(s,1H),3.76(t,2H),3.59(d,1H),3.35(d,2H),3.08–2.83(m,4H),2.25-2.23(m,1H),2.16-2.10(m,1H),2.02-1.95(m,2H),1.75(s,4H),1.19-1.13(m,2H),0.29(s,4H).
使用与实施例9中类似的条件，制备了如下表中的化合物。
实施例10
4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)苯甲酰胺(化合物97)

第一步(2Z)-N-(2,6-二溴苯胺)氮杂卓-2-亚胺(化合物97-2)的合成：
氮气保护，0℃下向己内酰胺(1.36g,15.94mmol,2eq)的甲苯(10mL)溶液中，滴加三氯氧磷(6.1g,39.85mmol,5eq)，反应液0℃下反应2小时。随后加入2,6-二溴苯胺(2g,7.97mmol,1eq)的甲苯溶液(5mL)，反应液升温至110℃，搅拌16小时。反应液冷却至室温，反应液减压浓缩，加冰水(50mL)淬灭，用饱和碳酸氢钠溶液调节pH值至10，混合物用乙酸乙酯萃取(3×60mL)。合并有机相，用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(10:1)，得到(2Z)-N-(2,6-二溴苯胺)氮杂卓-2-亚胺(2.7g,27.65％)。
MS:(ESI,m/z):345.10[M+H]+,RT(min):0.595.
第二步4-溴-7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓(化合物97-3)的合成：
氮气保护，室温下向(2Z)-N-(2,6-二溴苯胺)氮杂卓-2-亚胺(500mg,1.445mmol,1eq)的乙腈(3mL)溶液中加入N,N-二甲基乙二胺(25.47mg,0.289mmol,0.10eq)，碘化亚铜(27.52mg,0.145mmol,0.05eq)，碳酸钾(599.04mg,4.335mmol,3eq)，反应液升温至100℃，搅拌反应1小时。反应液冷却至室温，反应液用水(40mL)稀释，并用乙酸乙酯萃取(3×30mL)。合并有机相，用饱和食盐水反洗(1×40mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。得到4-溴-7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓(250mg,65.26％)。
MS:(ESI,m/z):264.90[M+H]+,RT(min):1.151.
第三步(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)氨基甲酸叔丁酯(化合物97-4)的合成：
氮气保护，室温下向4-溴-7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓(250mg,0.943mmol,1eq)的N,N-二甲基甲酰胺(10mL)中加入氨基甲酸叔丁酯(220.91mg,1.886mmol,2eq)，二环己基(3-异丙基-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦(50.42mg,0.094mmol,0.1eq)，(甲磺酸{双环乙基(3-异丙基-2,4,6-三异丙基-(1,1-联苯)-2-基)膦烷})(2-甲氨基-1,1-联苯-2-基)钯(Ⅱ)(173.21mg,0.189mmol,0.2eq)和碳酸铯(921.59mg,2.829mmol,3eq)。反应液升温至100℃，搅拌反应1小时。反应冷却至室温，用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)氨基甲酸叔丁酯(150mg,52.79％)。
MS:(ESI,m/z):302.35[M+H]+,RT(min):1.106.
第四步7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-胺(化合物97-5)的合成：
室温下向(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)氨基甲酸叔丁酯(150mg,0.498mmol,1eq)的二氯甲烷(2mL)溶液中，加入盐酸的1,4-二氧六环溶液(4M，2mL)。反应液室温搅拌1小时。反应液减压浓缩，得到7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-胺(150mg,66.67％)。未纯化，直接投下一步反应。
MS:(ESI,m/z):202.40[M+H]+,RT(min):0.544.
第五步4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)苯甲酰胺(化合物97-6)的合成：
氮气保护，室温下向7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-胺(150mg,1.443mmol,1eq)，2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(319.44mg,0.894mmol,1.2eq)和N,N,N,N-四甲基氯甲脒六氟磷酸盐(836.42mg,2.980mmol,4eq)的二氯甲烷(5mL)溶液中加入N-甲基咪唑(611.90mg,7.450mmol,10eq)。反应液室温搅拌2小时。用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水反洗(1×20mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到4-碘-2-(6-氮 杂螺[2.5]辛烷-6-基)-N-(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)苯甲酰胺(60mg,14.90％)。
MS:(ESI,m/z):541.35[M+H]+,RT(min):1.398.
第六步4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)苯甲酰胺(化合物97)的合成：
氮气保护，室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)苯甲酰胺(60mg,0.117mmol,1eq)和2-羟基乙磺酰胺(17.58mg,0.140mmol,1.2eq)的N,N-二甲基甲酰胺(2mL)溶液中，依次加入肌氨酸(2.09mg,0.023mmol,0.2eq)，碘化亚铜(2.23mg,0.012mmol,0.1eq)和碳酸钾(48.55mg,0.351mmol,3eq)。反应液升温至120℃，搅拌反应1小时。反应液冷却至室温，用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水溶液反洗(1×10mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得粗品用制备型高压液相纯化，得到4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(7,8,9,10-四氢-6H-苯并[4,5]咪唑并[1,2-a]氮杂卓-4-基)苯甲酰胺(12.53mg,22.81％)。
MS:(ESI,m/z):538.00[M+H]+,RT(min):1.588.
1H NMR(400MHz,DMSO-d6)δ12.47(s,1H),10.15(s,1H),8.30(d,1H),8.03(s,1H),7.27(d,1H),7.23(d,1H),7.16(d,1H),7.09(s,1H),4.94(s,1H),4.27(d,2H),3.76(s,2H),3.37(s,2H),3.09(s,2H),3.01-2.93(m,4H),1.90(s,4H),1.73(s,6H),0.30(s,4H).
实施例11
4-(2-羟基乙磺酰氨基)-N-(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物98)
第一步(2Z)-N-(2,6-二溴苯基)-3-甲基哌啶-2-亚胺(化合物98-2)的合成：
氮气保护，0℃下，向3-甲基哌啶-2-酮(1.62g,14.347mmol,1.2eq)的甲苯溶液(20mL)中，加入三氯氧磷(3.67g,23.912mmol,2eq)。加入完毕后反应体系在0℃下继续搅拌2小时。然后加入2,6-二溴苯胺(3g,11.956mmol,1eq)，加入完毕后反应体系在110℃下继续搅拌1小时。反应混合物冷却至室温，用水淬灭(20mL)。反应混合物用乙酸乙酯萃取(3×60mL)。合并有机相，用饱和氯化钠溶液反洗(1×60mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到(2Z)-N-(2,6-二溴苯基)-3-甲基哌啶-2-亚胺(2.3g，55.59％)。
MS(ESI,m/z):346.85[M+H]+,RT(min):0.620
第二步6-溴-4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶(化合物98-3)的合成：
氮气保护，室温下向(2Z)-氮-(2,6-二溴苯基)-3-甲基哌啶-2-亚胺(1.3g,3.757mmol,1eq)和碳酸钾(1038.34mg,7.514mmol,2eq)的乙腈溶液(2mL)中加入碘化亚铜(71.54mg,0.376mmol,0.1eq)和N,N-二甲基乙二胺(66.23mg,0.751mmol,0.2eq)。加入完毕后反应体系升温至100℃，继续搅拌1小时。反应混 合物冷却至室温，用水淬灭(10mL)。反应混合物用乙酸乙酯萃取(1×20mL)。合并有机相，用饱和氯化钠溶液反洗(1×20mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。粗品/所得混合物未进一步纯化，直接投下步。
MS(ESI,m/z):264.90[M+H]+,RT(min):0.561
第三步(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(化合物98-4)的合成：
氮气保护，室温下向6-溴-4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶(950mg,3.583mmol,1eq)和氨基甲酸叔丁酯(503.66mg,4.300mmol,1.2eq)的N,N-二甲基甲酰胺(10mL)溶液中，加入碳酸铯(3.50g,10.749mmol,3eq)，双环已基(3-异丙氧-2′,4′,6′-三异丙基-[1,1′-联苯]-2-基)膦烷(383.23mg,0.717mmol,0.2eq)和甲磺酸{双环己基(3-异丙氧-2',4′,6′-三异丙基-[1,1′-联苯]-2-基)膦烷}(2'-甲氨基-1,1'-联苯-2-基)钯(II)(329.11mg,0.358mmol,0.1eq)，加入完毕后反应体系升温至100℃，继续搅拌1小时。反应液冷却至室温。反应混合物用乙酸乙酯萃取(3×30mL)。合并有机相，用饱和氯化钠溶液反洗(1×40mL)，无水硫酸钠干燥。所得混合物过滤后，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(950mg,87.98％)。
MS(ESI,m/z):302.30[M+H]+,RT(min):1.128
第四步4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(化合物98-5)的合成：
氮气保护，室温下向(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(700mg,2.323mmol,1eq)的二氯甲烷溶液(7mL)中加入氯化氢的1,4-二氧六环溶液(7.5mL，4mol/L)，反应液室温继续搅拌1小时。反应液减压浓缩。所得残余物未进一步纯化，直接投下步。
MS(ESI,m/z):202.25[M+H]+,RT(min):0.777
第五步4-碘-N-(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物98-6)的合成：
氮气保护，室温下向4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(520mg,2.584mmol,1eq)和2-(6-氮杂螺[2.5]辛烷-6-基-4-碘苯甲酸(1.11g,3.101mmol,1.2eq)的二氯甲烷溶液(10mL)中，加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(2.90g,10.336mmol,4eq)和N-甲基咪唑(2.12g,25.840mmol,10eq)。加入完毕后，反应体系在室温下继续搅拌1小时。反应混合物用乙酸乙酯萃取(3×40mL)。合并有机相，用饱和氯化钠溶液反洗(1×50mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(4:1)，得到4-碘-N-(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(620mg,44.40％)。
MS(ESI,m/z):541.25[M+H]+,RT(min):1.585
第六步4-(2-羟基乙磺酰氨基)-N-(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物98)的合成：
氮气保护，室温下向4-碘-N-(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(100mg,0.185mmol,1eq)和2-羟基乙磺酰胺(27.79mg,0.222mmol,1.2eq)的N,N-二甲基甲酰胺溶液(2mL)中，加入2-(甲氨基)乙酸(13.19mg,0.148mmol,0.8eq)，碘化亚铜(14.1mg,0.074mmol,0.4eq)和磷酸钾(117.83mg,0.555mmol,3eq)，加入完毕后，反应体系升温至120℃，继续搅拌1.5小时。反应液冷却至室温，用乙酸乙酯萃取(3×10mL)。合并有机相，用饱和氯化钠溶液反洗(1×20mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。粗品通过制备型高效液相纯化，条件如下：层析柱规格:Kinetex 5μm EVO C18,30mm*150mm；流动相A:水(10mmol/L碳酸氢钠),流动相B:乙腈；流速:60ml/min；洗脱梯度:10％B to 37％B in 10min；检测波长:254nm/220nm；保留时间(min):9.03，得到4-(2-羟基乙磺酰氨基)-N-(4-甲基-1,2,3,4-四氢苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(35.6mg,35.39％)。
MS(ESI,m/z):538.40[M+H]+,RT(min):1.536
1H NMR(400MHz,DMSO-d6)δ12.07(s,1H),10.12(s,1H),8.39(dd,1H),8.01(d,1H),7.25(d,1H),7.21–7.10(m,2H),7.07(dd,1H),5.11(d,1H),4.25–4.11(m,1H),4.10–3.93(m,1H),3.76(t,2H),3.35(t,2H),3.20–3.07(m,1H),3.02(s,4H),2.24–2.07(m,2H),2.08–1.94(m,1H),1.93–1.65(m,4H),1.68–1.56(m,1H),1.48(d,3H),0.30(s,4H).
使用与实施例11中类似的条件，制备了如下表中的化合物。


实施例12
4-(2-羟基乙磺酰氨基)-N-((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物99)
第一步(1S,4R)-2-(3-氯-5-甲基-2-硝基苯基)-2-氮杂双环[2.2.1]庚-3-酮(化合物99-3)的合成：
氮气保护，室温下向1,3-二氯-5-甲基-2-硝基苯(1g,4.854mmol,1eq)，磷酸钾(3.09g,14.562mmol,3eq)和(1S,4R)-2-氮杂双环[2.2.1]庚-3-酮(539.48mg,4.854mmol,1eq)的1,4-二氧六环溶液(10mL)中加入醋酸钯(108.97mg,0.485mmol,0.1eq)，1,1'-双(二苯基膦)二茂铁(536.23mg,0.971mmol,0.2eq)，反应液升温至100℃搅拌反应1小时。反应液冷却到室温，过滤，滤饼用乙酸乙酯洗涤(3×10mL)，反应混合物用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和食盐水反洗(2×50mL)，无水硫酸钠干燥。所得混合物过滤后，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(1:1)，得到(1S,4R)-2-(3-氯-5-甲基-2-硝基苯基)-2-氮杂双环[2.2.1]庚-3-酮(800mg,58.71％)。
MS:(ESI,m/z):281.30[M+H]+,RT(min):1.079
第二步(1S,4R)-6-氯-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶(化合物99-4)的合成：
氮气保护，室温下向(1S,4R)-2-(3-氯-5-甲基-2-硝基苯基)-2-氮杂双环[2.2.1]庚-3-酮(790mg,2.814mmol,1eq)的乙醇(10mL)溶液中加入还原铁粉(785.82mg,14.070mmol,5eq)，醋酸(4mL)。反应液升温至100℃继续搅拌2小时。反应液冷却到室温，减压浓缩。粗品用水(50mL)稀释后，混合物用乙酸乙酯萃取(3×40mL)。合并有机相，用饱和食盐水反洗(2×30mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(1:1)，得到(1S,4R)-6-氯-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶(560mg,85.51％)。
MS:(ESI,m/z):233.35[M+H]+,RT(min):0.814
第三步((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(化合物99-5)的合成：
氮气保护，室温下向(1S,4R)-6-氯-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶(550mg,2.363mmol,1eq)，碳酸铯(2.31g,7.089mmol,3eq)和氨基甲酸叔丁酯(553.75mg,4.726mmol,2eq)的N,N-二甲基甲酰胺溶液(6mL)中加入(甲磺酸{双环己基(3-异丙氧-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦烷}(2'-甲氨基-1,1'-联苯-2-基)钯(II)4(217.10mg,0.236mmol,0.1eq)，二环己基(3-异丙氧基-2',4',6'-三异丙基-[1,1'-联苯基]-2-基)磷烷(252.80mg,0.473mmol,0.2eq)，反应液升温至100℃，搅拌16小时。反应液冷却到室温，用乙酸乙酯萃取(3×40mL)。合并有机相，用饱和食盐水反洗(2×30mL)，无水硫酸钠干燥。所得混 合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(1:1)，得到((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(650mg,87.75％)。
MS:(ESI,m/z):314.45[M+H]+,RT(min):0.871
第四步(1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(化合物99-6)的合成：
室温下向((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(650mg,2.074mmol,1eq)的二氯甲烷(10mL)溶液中加入氯化氢的1,4-二氧六环溶液(10mL)，搅拌反应1小时。液质监测反应完全，将混合物真空浓缩，得到(1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺，粗品(700mg)，粗品未进一步纯化，直接投下步。
MS:(ESI,m/z):214.40[M+H]+,RT(min):0.230
第五步4-碘-N-((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物99-7)的合成：
氮气保护，室温下向(1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(107.48mg,0.504mmol,1.2eq)，4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酸(150mg,0.420mmol,1.0eq)和N,N,N',N'-四甲基氯甲脒六氟磷酸盐(471.31mg,1.680mmol,4eq)的二氯甲烷溶液(5mL)中加入N-甲基咪唑(344.80mg,4.200mmol,10eq)。反应液室温下继续搅拌3小时。反应液用二氯甲烷萃取(3×30mL)。合并有机相，用饱和食盐水反洗(2×30mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(2:1)，得到4-碘-N-((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(110mg,47.41％)。
MS:(ESI,m/z):553.10[M+H]+,RT(min):1.118
第六步4-(2-羟基乙磺酰氨基)-N-((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物99)的合成：
氮气保护，室温下向4-碘-N-((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(100mg,0.181mmol,1eq)，碳酸钾(75.05mg,0.543mmol,3eq)和2-羟基乙烷-1-磺酰胺(45.30mg,0.362mmol,2eq)的N,N-二甲基甲酰胺溶液(2mL)中加入碘化亚铜(3.45mg,0.018mmol,0.1eq)，肌氨酸(3.23mg,0.036mmol,0.2eq)，反应液升温至120℃，搅拌反应2小时。反应液冷却到室温，过滤，滤饼用乙酸乙酯洗涤(3×5mL)，混合物用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水反洗(2×30mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。粗品通过制备型高效液相纯化，条件如下(层析柱规格:Kinetex 5μm EVO C18,30mm×150mm；流动相A:水(10mmol/L碳酸氢铵),流动相B:乙腈；流速:60ml/min；洗脱梯度:27％B to 62％B in 8min；检测波长:254nm/220nm；保留时间(分钟):7.92)，得到4-(2-羟基乙磺酰氨基)-N-((1S,4R)-8-甲基-1,2,3,4-四氢-1,4-亚甲基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(38.29mg,38.33％)，
MS:(ESI,m/z):550.45[M+H]+,RT(min):1.525
1H NMR(400MHz,DMSO-d6)δ12.22(s,1H),10.13(s,1H),8.15(d,1H),7.98(d,1H),7.23(d,1H),7.10–7.01(m,2H),5.09(s,1H),4.94(s,1H),3.76(t,2H),3.55(d,1H),3.35(d,2H),2.97(d,4H),2.41(s,3H),2.21(d,1H),2.14–2.05(m,1H),2.00–1.90(m,2H),1.72(s,4H),1.20–1.07(m,2H),0.28(s,4H).
使用与实施例12中类似的条件，制备了如下表中的化合物。
实施例13
4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(化合物100)
第一步2-(3-溴-2-硝基苯基)-2-氮杂双环[2.2.2]辛烷-3-酮(化合物100-3)的合成：
氮气保护，室温下向1,3-二溴-2-硝基苯(1g,3.56mmol,1eq)，2-氮杂双环[2.2.2]辛烷-3-酮(534mg,4.3mmol,1.2eq)和碳酸铯(3.5g,10.7mmol,3eq)的1,4-二氧六环溶液(10mL)中加入三(二亚苄基丙酮)二钯(326mg,0.356mmol,0.1eq),4,5-双二苯基膦-9,9-二甲基氧杂蒽(412mg,0.712mmol,0.2eq)，加入完毕后反应液升温至80℃，搅拌反应3小时。反应液冷却至室温，过滤，滤饼用乙酸乙酯洗涤(3×10mL)，混合物用乙酸乙酯萃取(3×50mL)，合并有机相，并用饱和食盐水反洗(2×50mL)，无水硫酸钠干燥，过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(1:1)，得到2-(3-溴-2-硝基苯基)-2-氮杂双环[2.2.2]辛烷-3-酮(603mg,52％)。
MS:(ESI,m/z):325.30[M+H]+,RT(min):1.021
第二步6-溴-1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶(化合物100-4)的合成：
氮气保护，室温下向2-(3-溴-2-硝基苯基)-2-氮杂双环[2.2.2]辛烷-3-酮(580mg,1.8mmol,1eq)的N,N-二甲基甲酰胺(1mL)溶液中，加入四羟基二硼(480mg,5.4mmol,3eq)，4,4'-联吡啶(1.4mg,0.009mmol,0.005eq)，加入完毕后室温下搅拌反应16小时。反应液室温下加水(10mL)淬灭，混合物用乙酸乙酯萃取(3×40mL)。合并有机相，并用饱和食盐水反洗(2×10mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(1:1)，得到6-溴-1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶(380mg,76％)。
MS:(ESI,m/z):277.20[M+H]+,RT(min):0.683
第三步(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(化合物100-5)的合成：
氮气保护，室温下向6-溴-1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶(100mg,0.36mmol,1eq)的1,4-二氧六环溶液(2mL)中加入氨基甲酸叔丁酯(51mg,0.43mmol,1.2eq)，(甲磺酸{双环己基(3-异丙氧-2',4',6'-三异丙基-[1,1'-联苯]-2-基)膦烷}(2'-甲氨基-1,1'-联苯-2-基)钯(II)4(33mg,0.036mmol,0.1eq)，二环己基(3-异丙氧基-2',4',6'-三异丙基-[1，1'-联苯基]-2-基)磷烷(39mg,0.072mmol,0.2eq)，碳酸铯(588mg,1.8mmol,5eq)，加入完毕后升温至100℃搅拌16小时。反应液冷却至室温，反应液过滤，滤饼用乙酸乙酯洗涤(3×5mL)，混合物用乙酸乙酯萃取(3×20mL)，合并有机相，并用饱和食盐水反洗(2×30mL)，无水硫酸钠干燥，过滤，滤液减压浓缩，所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(1:1)，得到(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(90mg,79％)。
MS:(ESI,m/z):314.40[M+H]+,RT(min):0.768
第四步1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(化合物100-6)的合成：
氮气保护,室温下向(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)氨基甲酸叔丁酯(85mg,0.27mmol,1eq)的二氯甲烷溶液(1mL)中，加入氯化氢的1,4-二氧六环溶液(1mL)，加料完毕后室温搅拌1小时。反应液减压浓缩，得到1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(57mg,98％)。
MS:(ESI,m/z):214.4[M+H]+,RT(min):0.196
第五步4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(化合物100-7)的合成：
氮气保护下，室温下向1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-胺(52mg,0.24mmol,1eq)的二氯甲烷溶液中(1mL)中，加入2-{6-氮杂螺[2.5]辛烷-6-基}-4-碘苯甲酸(104mg,0.23mmol,1.2eq)，N,N,N',N'-四甲基氯甲脒六氟磷酸盐(342mg,1.2mmol,5eq)，N-甲基咪唑(200mg,2.4mmol,10eq)，加料完毕后室温搅拌反应1小时。反应液用乙酸乙酯萃取(3×30mL)，合并有机相，并用饱和食盐水反洗(2×20mL)，无水硫酸钠干燥。过滤，滤液减压浓缩，所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(3:1)，得到4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(110mg,82％)。
MS:(ESI,m/z):553.5[M+H]+,RT(min):1.235
第六步4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(化合物100)的合成：
氮气保护，室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(100mg,0.18mmol,1eq)的N’,N’-二甲基甲酰胺(100mg,0.18mmol,1eq)溶液中，加入2-羟基乙烷磺酰胺(27mg,0.2mmol,1.2eq)，碘化亚铜(3.5mg,0.018mmol,0.1eq)，肌氨酸(3.2mg,0.036mmol,0.2eq)和碳酸钾(75mg,0.5mmol,3eq)。反应液120℃下搅拌1.5小时。反应液冷却至室温，反应液过滤，滤饼用乙酸乙酯洗涤(3×5mL)，混合物用乙酸乙酯萃取(3×30mL)，合并有机相，并用饱和食盐水反洗(2×20mL)，无水硫酸钠干燥。过滤，滤液减压浓缩，所得残余物用制备型高效液相纯化，条件如下：层析柱规格:Kinetex 5μm EVO C18,30mm×150mm；流动相A:水(10mmol/L碳酸氢钠),流动相B:乙腈；流速:60ml/min；洗脱梯度:30％B to 60％B in 8min；检测波长:254nm/220nm；保留时间(min):7.65，得到4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)-N-(1,2,3,4-四氢-1,4-乙基苯并[4,5]咪唑并[1,2-a]吡啶-6-基)苯甲酰胺(28.84mg,28％)。
MS:(ESI,m/z):550.10[M+H]+,RT(min):1.548
1H NMR:(400MHz,CDCl3)δ12.28(s,1H),8.34(d,1H),8.01(d,1H),7.85(d,1H),7.64–7.28(m,1H),7.27(s,1H),7.15(d,1H),6.92(dd,1H),4.83(s,1H),4.05(t,2H),3.52(s,2H),3.27(t,2H),3.08(t,4H),2.02(dt,4H),1.88–1.66(m,8H),0.24(s,4H).
实施例14
N-(3,3-二氟-2,3-二氢-1H-苯并吡咯并[1,2-a]咪唑-5-基)-4-(2-羟基乙磺酰氨基)-2-螺[2.5]辛-5-烯-6-基)苯甲酰胺(化合物90)

第一步4-溴-2-(螺[2.5]辛-5-烯-6-基)苯甲酸甲酯(化合物90-3)的合成：
氮气保护，室温下向4-溴-2-碘苯甲酸甲酯(700mg,2.053mmol,1eq)，碳酸钠(326.41mg,3.079mmol,1.5eq)和4,4,5,5-四甲基-2-{螺[2.5]辛-5-烯-6-基}-1,3,2-二氧硼杂环戊烷(480.74mg,2.053mmol,1eq)的1,4-二氧六环(8mL)和水(1.6mL)的溶液中加入[1,1-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(167.25mg,0.205mmol,0.1eq)。反应液升温至80℃,继续搅拌1小时。反应液冷却至室温。反应液过滤，滤饼用乙酸乙酯洗涤(3×50mL)。滤液用水稀释(30mL)。混合物用乙酸乙酯萃取(2×50mL)，合并有机相，并用饱和食盐水反洗(1×50mL)，无水硫酸钠干燥。过滤，滤液减压浓缩。所得残余物用反相柱纯化，条件如下：层析柱规格,C18；流动相:乙腈in水,梯度10％to 70％in 10min；检测波长:UV 254nm.得到4-溴-2-(螺[2.5]辛-5-烯-6-基)苯甲酸甲酯(320mg,48.52％)。
MS:(ESI,m/z):320.95[M+H]+,RT(min):1.942
第二步4-溴-2-{螺[2.5]辛-5-烯-6-基}苯甲酸(化合物90-4)的合成：
室温下，向甲基4-溴-2-(螺[2.5]辛-5-烯-6-基)苯甲酸酯(300mg,0.934mmol,1eq)的甲醇(2mL)，四氢呋喃(2mL)和水(2mL)的溶液中加入氢氧化锂(111.84mg,4.670mmol,5eq)。反应液室温搅拌16小时。反应液用水稀释(15mL)，混合物用乙酸乙酯萃取(2×20mL)，收集水相，并用1摩尔每升的稀盐酸酸化至pH 4～5，水相用乙酸乙酯萃取(3×30mL)。合并收集有机相，并用饱和食盐水反洗(2×20mL)，无水硫酸钠干燥。过滤，滤液减压浓缩得到4-溴-2-{螺[2.5]辛-5-烯-6-基}苯甲酸(250mg,87.14％)。
MS:(ESI,m/z):308.90[M+H]+,RT(min):1.323
第三步4-溴-N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-2-(螺[2.5]辛-5-烯-6-基)苯甲酰胺(化合物90-5)的合成：
在氮气保护，室温下向3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-胺(112.37mg,0.537mmol,1.1eq)，N,N,N’,N’-四甲基氯甲脒六氟磷酸盐(548.03mg,1.952mmol,4eq)和4-溴-2-(螺[2.5]辛-5-烯-6-基)苯甲酸(150mg,0.488mmol,1.00eq)的二氯甲烷(5mL)溶液中，滴加N-甲基咪唑(400.93mg,4.880mmol,10eq)。加入完毕后体系60℃下继续搅拌2小时。反应液冷却到室温，用乙酸乙酯萃取(3×30mL)。合并有机相，用饱和食盐水反洗(2×20mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(2:1)，得到4-溴-N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-2-(螺[2.5]辛-5-烯-6-基)苯甲酰胺(64mg,26.30％)。
MS:(ESI,m/z):498.40[M+H]+,RT(min):1.449
第四步4-溴-N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-2-(螺[2.5]辛-5-烯-6-基)苯甲酰胺(化合物90)的合成：
氮气保护，室温下向4-溴-N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-2-(螺[2.5]辛-5-烯-6-基)苯甲酰胺(50mg,0.100mmol,1eq)，碳酸铯(98.06mg,0.300mmol,3eq)和2-羟基乙烷-1-磺酰胺(25.11mg,0.200mmol,2eq)的N,N-二甲基甲酰胺(2mL)溶液中加入(甲磺酸{双环己基(3-异丙氧-2’,4’,6’-三异丙基-[1,1’-联苯]-2-基)膦烷}(2’-甲氨基-1,1’-联苯-2-基)钯(II)(9.22mg,0.010mmol,0.1eq)，双环已基(3-异丙氧-2’,4’,6’-三异丙基-[1,1’-联苯]-2-基)膦烷(10.73mg,0.020mmol,0.2eq)反应液升温至100℃，搅拌反应1小时。反应液冷却到室温，用乙酸乙酯萃取(3×30mL)，合并有机相，用饱和食盐水反洗(1×30mL)，无水硫酸钠干燥。所得混合物过滤后，将滤液减压浓缩。粗品通过高效液相纯化，条件如下(层析柱规格: Kinetex 5m EVO C18,30mm*150mm；流动相A:水(10毫摩尔每升碳酸氢铵)，流动相B:乙腈；流速:60毫升每分钟；洗脱梯度:23％B升至60％B用时8分钟；检测波长:UV 254nm/220nm；保留时间(分钟):7.6)。得到N-(3,3-二氟-2,3-二氢-1H-苯并[d]吡咯并[1,2-a]咪唑-5-基)-4-(2-羟基乙磺酰氨基)-2-(螺[2.5]辛-5-烯-6-基)苯甲酰胺(10.84mg,19.83％)，
MS:(ESI,m/z):543.50[M+H]+,RT(min):1.650
1H NMR(400MHz,DMSO-d6)δ9.92(s,1H),9.48(s,1H),8.13(d,1H),7.59(d,1H),7.34–7.29(m,1H),7.26(t,1H),7.14(dd,1H),6.99(d,1H),5.88(s,1H),4.84(s,1H),4.34(d,2H),3.66(t,2H),3.16(d,4H),2.16(s,2H),2.08–1.98(m,2H),1.16(t,2H),0.00(d,4H).
使用与实施例14中类似的条件，制备了如下表中的化合物。
实施例15
N-(二苯并[b,d]呋喃-4-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物111)
第一步N-(二苯并[b,d]呋喃-4-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物111-2)的合成：
氮气保护，室温下向4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酸(100mg,0.280mmol,1eq)的二氯甲烷(2mL)溶液中加入4-氨基氧芴(61.55mg,0.336mmol,1.2eq)，N,N,N’,N’-四甲基氯甲脒六氟磷酸盐(314.21mg,1.120mmol,4eq)和N-甲基咪唑(229.87mg,2.800mmol,10eq)，反应液升温至80℃，搅拌反应1小时。反应液冷却至室温，反应混合物加水(20mL)淬灭。反应混合物用乙酸乙酯萃取(3×30mL)。合并有机相，用饱和食盐水反洗(3×20mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(10:1)，得到N-(二苯并[b,d]呋喃-4-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(140mg,95.73％)。
MS:(ESI,m/z):523.30[M+H]+,RT(min):1.722
第二步N-(二苯并[b,d]呋喃-4-基)-4-((2-羟乙基)磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(化合物111)的合成：
氮气保护，室温下向N-(二苯并[b,d]呋喃-4-基)-4-碘-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(90mg,0.172mmol,1eq)的N,N-二甲基甲酰胺(2mL)溶液中加入2-羟基乙烷-1-磺酰胺(32.34mg,0.258mmol,1.5eq)，碘化亚铜(8.2mg,0.043mmol,0.25eq)，碳酸钾(71.43mg,0.516mmol,3eq)和2-(甲氨基)乙酸(6.14mg,0.069mmol,0.4eq)，反应液升温至120℃，搅拌反应1小时。反应液冷却至室温，用水淬灭。反应混合物用乙酸乙酯萃取(3×20mL)。合并有机相，用饱和食盐水反洗(2×20mL)，无水硫酸钠干燥。所得混合物过滤，将滤液减压浓缩。粗品通过高效液相纯化，条件如下：层析柱规格:Kinetex EVO prep C18,30*150,5μm；流动相A:水(10mmol/mL碳酸氢钠)，流动相B:乙腈；流速:60mL/min；洗脱梯度:45％B升至70％B in 8min；检测波长:UV 220nm；保留时间(min):7.03。得到N-(二苯并[b,d]呋喃-4-基)-4-(2-羟基乙磺酰氨基)-2-(6-氮杂螺[2.5]辛烷-6-基)苯甲酰胺(25.9mg,27.89％)。
MS+:(ESI,m/z):519.90[M+H]+,RT(min):1.784.
1H NMR:(400MHz,DMSO-d6)δ12.85(s,1H),10.21(s,1H),8.74–8.62(m,1H),8.26–8.18(m,1H),8.17–8.07(m,1H),7.95–7.84(m,1H),7.73(d,1H),7.67–7.57(m,1H),7.51–7.45(m,1H),7.44–7.35(m,2H),7.23–7.13(m,1H),4.95(s,1H),3.84–3.72(m,2H),3.40–3.35(m,2H),3.14–2.97(m,4H),1.83(s,4H),0.43(d,4H).
实施例16
2-{6-氮杂螺[2.5]辛烷-6-基}-N-(7-环丙基喹啉-4-基)-4-(2-羟基乙磺酰氨基)苯甲酰胺(化合物112)

第一步N-(7-溴喹啉-4-基)氨基甲酸叔丁酯(化合物112-2)的合成：
在室温下，向7-溴喹啉-4-胺(1.0g,4.483mmol,1eq)的二氯甲烷(15.0mL)溶液中加入三乙胺(1.36g,13.449mmol,3eq)和二碳酸二叔丁酯(978.37mg,4.483mmol,1eq)。反应液室温下搅拌17小时。还有原料剩余，反应体系加热至50℃反应1小时。反应液冷却至室温，反应混合物用二氯甲烷萃取(3×60mL)。合并有机相，用饱和食盐水反洗(2×60mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到N-(7-溴喹啉-4-基)氨基甲酸叔丁酯(1.16g,80.07％)。
MS:(ESI,m/z):323.05[M+H]+,RT(min):1.427
第二步N-(7-溴喹啉-4-基)氨基甲酸叔丁酯(化合物112-4)的合成：
氮气保护，室温下向N-(7-溴喹啉-4-基)氨基甲酸叔丁酯(700mg,2.166mmol,1eq)的1,4-二氧六环溶液(2mL)和水(0.5mL)的混合溶液中加入环丙基硼酸(558.15mg,6.498mmol,3eq)，[1,1'-双(二苯基膦)二茂铁]二氯化钯二氯甲烷络合物(176.44mg,0.217mmol,0.1eq)，碳酸铯(2117.08mg,6.498mmol,3eq)。反应液升温至80℃，搅拌一个小时。反应液冷却至室温，用乙酸乙酯萃取(3×50mL)。合并有机相，用饱和食盐水反洗(2×40mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(1:1)，得到N-(7-环丙基喹啉-4-基)氨基甲酸叔丁酯(530mg,86.05％)。
MS:(ESI,m/z):285.30[M+H]+,RT(min):1.054
第三步7-环丙基喹啉-4-胺(化合物112-5)的合成：
室温下向N-(7-环丙基喹啉-4-基)氨基甲酸叔丁酯(500mg,1.758mmol,1eq)的二氯甲烷(5mL)溶液中加入4摩尔每升氯化氢的1,4-二氧六环中(2.5mL)，搅拌反应30min。所得残余物减压浓缩，得到产物7-环丙基喹啉-4-胺(240mg,74.08％)。
MS:(ESI,m/z):185.20[M+H]+,RT(min):0.672
第四步2-(6-氮杂螺[2.5]辛烷-6-基-N-(7-环丙基喹啉-4-基)-4-碘苯甲酰胺(化合物112-6)的合成：
氮气保护，室温下向7-环丙基喹啉-4-胺(100mg,0.543mmol,1eq)，2-(6-氮杂螺[2.5]辛烷-6-基-4-碘苯甲酸(290.81mg,0.815mmol,1.5eq)的二氯甲烷(4mL)溶液中加入N,N,N',N'-四甲基氯甲脒六氟磷酸盐(609.16mg,2.172mmol,4eq)，N-甲基咪唑(445.64mg,5.430mmol,10eq)。反应液室温搅拌一个小时。反应混合物加入水(3mL)淬灭。反应混合物用二氯甲烷萃取(2×40mL)。合并有机相，用饱和食盐水反洗(2×20mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。所得残余物用硅胶柱层析纯化，石油醚/乙酸乙酯(5:1)，得到2-(6-氮杂螺[2.5]辛烷-6-基-N-(7-环丙基喹啉-4-基)-4-碘苯甲酰胺(70mg,24.64％)。
MS:(ESI,m/z):524.45[M+H]+,RT(min):1.029
第五步2-{6-氮杂螺[2.5]辛烷-6-基}-N-(7-环丙基喹啉-4-基)-4-(2-羟基乙磺酰氨基)苯甲酰胺(化合物112)的合成：
氮气保护，室温下向2-{6-氮杂螺[2.5]辛烷-6-基}-N-(7-环丙基喹啉-4-基)-4-碘苯甲酰胺(60mg,0.115mmol,1eq)的N,N-二甲基甲酰胺(1mL)溶液中加入2-(甲氨基)乙酸(2.04mg,0.023mmol,0.2eq)，碳酸钾(47.53mg,0.345mmol,3eq)，碘化亚铜(2.18mg,0.012mmol,0.1eq)和2-羟基乙烷-1-磺酰胺(17.21mg,0.138mmol,1.2eq)。反应液升温至120℃，搅拌1.5小时。反应液冷却至室温，用乙酸乙酯萃取(2×30mL)。合并有机相，用饱和食盐水反洗(2×20mL)，无水硫酸钠干燥。所得混合物过滤，滤液减压浓缩。粗品通过高效液相纯化，得到2-{6-氮杂螺[2.5]辛烷-6-基}-N-(7-环丙基喹啉-4-基)-4-(2-羟基乙磺酰氨基)苯甲酰胺(18.96mg,31.55％)。
MS:(ESI,m/z):521.40[M+H]+,RT(min):1.352
1H NMR(400MHz,DMSO-d6)δ11.99(s,1H),10.17(s,1H),8.76(d,,1H),8.33(d,1H),8.23(d,1H),7.90 (d,,1H),7.69(d,1H),7.39(dd,,1H),7.23(s,1H),7.06(d,1H),4.96(s,1H),3.78(t,2H),3.35(t,2H),3.03(d,4H),2.18(ddd,1H),1.41(s,4H),1.15–1.06(m,2H),0.91–0.83(m,2H),0.25(s,4H).
生物学评价
测试例1
测试名称：OVCAR-3细胞中基于成像的核计数分析(NCA)
第0天化合物稀释及处理
a)AM-5308最终测试浓度为：10000、3333.3、1111.1、370.3、123.4、41.1、13.7、4.5、1.5、0.5nM。
b)测试化合物，最终测试浓度为：10000、3333.3、1111.1、370.3、123.4、41.1、13.7、4.5、1.5、0.5nM。
c)细胞在37℃、5％CO2的培养箱中培养4天。
d)DMSO浓度为0.1％。
第1天将细胞接种到384孔细胞培养板中
a)当细胞汇合度达到80％-90％时，处理细胞。
b)将细胞重悬于培养基中，然后以所需的密度对细胞进行计数和稀释。
c)在384孔板中加入30μL/孔的含有适当细胞的细胞悬液：600个细胞/孔。
第4天检测
a)加入30μL 8％的固定液(终浓度4％)并在室温下孵育板30分钟。
b)离心板，1000RPM，30s。
c)60μl/孔PBS洗涤两次。
d)固定后，将细胞透化并在含有2μg/mL Hoechst 33342 DNA染料的60μL洗涤缓冲液中染色(1％BSA、0.2％Triton X-100、1X PBS)。
e)将板密封并在室温下避光孵育1小时。
f)用PBS洗涤3次。
g)加入50μLPBS/孔，并使用HCS扫描板子。
h)数据采集与检测
数据分析
抑制率(％)＝100-(化合物孔读值-低读值对照孔读值)/(高读值对照孔读值-低读值对照孔读值)*100
高读值对照孔：细胞加30nL DMSO；低读值对照孔：10μM AM-5308孔。
使用GraphPad Prism 8软件计算IC50(nM)并绘制化合物的效果-剂量曲线。
表1本申请化合物的生物学活性数据

测试例2
测试名称：KIF18A酶活检测
操作步骤：
1)化合物稀释及处理：AM-5308最终测试浓度为：10000、3333.3、1111.1、370.3、123.4、41.1、13.7、4.5、1.5、0.5nM，测试化合物最终测试浓度为：10000、3333.3、1111.1、370.3、123.4、41.1、13.7、4.5、1.5、0.5nM。
2)用Echo 655向反应板每孔转移100nL稀释好的化合物储液。DMSO的最终浓度为1％。
3)用封板膜封住反应板，1000g离心1分钟。
4)用1×反应缓冲液配制2×酶溶液。
5)向反应板中每孔加入5μL 2×酶溶液。用封板膜封住板子1000g离心1分钟，室温放置15分钟。
6)用1×反应缓冲液配制2×ATP溶液。
7)向反应板中加入5μL 2×ATP溶液，1000g离心1分钟，开始反应。
8)室温反应60分钟。
9)加入10μL ADP Glo试剂。1000g离心1分钟，在室温下孵育60分钟。
10)加入20μL激酶检测试剂。1000g离心1分钟，室温下孵育60分钟。
11)1000g离心1分钟。
12)在Envision 2104读取发光信号。
数据分析：
抑制百分率计算如下：
％inhibition＝100-(Signalcmpd-SignalAve_PC)/(SignalAve_VC-SignalAve_PC)×100
Signalcmpd:反应板上测试化合物的平均值。
SignalAve_PC:反应板上阳性对照(AM-5308)的平均值。
SignalAve_VC:反应板上阴性对照(DMSO)的平均值。
计算IC50以及拟合化合物量效曲线:
用GraphPad 8.0，利用非线性拟合公式来得到化合物的IC50。
3)质量控制
Z factor>0.5；S/B>2。
表2本申请化合物的酶活数据
以上对本发明技术方案的实施方式进行了示例性的说明。应当理解，本发明的保护范围不拘囿于上述实施方式。凡在本发明的精神和原则之内，本领域技术人员所做的任何修改、等同替换、改进等，均应包含在本申请权利要求书的保护范围之内。

Claims (10)

  1. 一种式(I)所示的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物：
    其中，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团包含两个、三个或四个彼此独立地选自饱和或部分不饱和的C3-14碳环、C6-14芳环、5-14元杂芳环、3-14元杂环的环；
    每个Ra相同或不同，彼此独立地选自H、OH、卤素、氰基、NH2、NO2，无取代或任选被一个、两个或更多个Ra1取代的下列基团：C1-12烷基、C1-12烷氧基、C3-12环烷基；或者，连接在同一个环碳原子上的两个Ra与其连接的碳原子一起共同形成饱和或部分不饱和的C3-14碳环；或者，两个不相邻的Ra以其端基相连，共同形成C1-3的亚烷基；每个Ra1相同或不同，彼此独立地选自H、OH、卤素、氰基、NH2、NO2、C1-12烷基、C1-12烷氧基、C3-12环烷基；
    E选自无取代或任选被一个、两个或更多个Re取代的下列基团：-NH-S(＝O)2-Re1、-S(＝O)2-NH-Re2、-S(＝O)(＝NH)-Re3、-N(Re4)(Re5)、3-14元杂环基；每个Re相同或不同，彼此独立地选自OH、卤素、氰基、C1- 12烷基、C1-12烷氧基、卤代C1-12烷基、卤代C1-12烷氧基、氰基C1-12烷基、氰基C1-12烷氧基、-N(Re6)(Re7)；
    每个Re1、Re2、Re3、Re4、Re5、Re6、Re7相同或不同，彼此独立地选自H、C1-12烷基、羟基C1-12烷基、卤代C1-12烷基、卤代C1-12烷氧基、氰基C1-12烷基、氰基C1-12烷氧基、C3-12环烷基、3-14元杂环基、C1-12烷氧基-C1-12烷基；
    M选自无取代或任选被一个、两个或更多个Rm取代的C3-12环烷基、C3-12环烯基或含氮的3-14元杂环基；每个Rm相同或不同，彼此独立地选自H、卤素、氰基、C1-12烷基、卤代C1-12烷基、氰基C1-12烷基、C1-12烷氧基、氰基C1-12烷氧基；或者，连接在同一个环碳原子上的两个Rm与其连接的碳原子一起共同形成饱和或部分不饱和的C3-14碳环；
    Y1、Y2、Y3相同或不同，彼此独立地选自N或CH。
  2. 根据权利要求1所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物，其特征在于，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团包含两个、三个或四个彼此独立地选自饱和或部分不饱和的C3-8碳环、C6-10芳环、5-10元杂芳环、3-8元杂环的环；
    每个Ra相同或不同，彼此独立地选自H、OH、卤素、氰基、NH2、NO2、C1-6烷基、C1-6烷氧基、C3-8环烷基、卤代C1-6烷基、卤代C1-6烷氧基、氰基C1-6烷基、氰基C1-6烷氧基、C3-8环烷基-C1-6烷氧基；或者，连接在同一个环碳原子上的两个Ra与其连接的碳原子一起共同形成饱和或部分不饱和的C3-8碳环；或者，两个不相邻的Ra以其端基相连，共同形成C1-3的亚烷基；
    优选地，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团选自、
    其中，T选自CH2、CH、NH、NRa或O；
    Z选自CH2、CH、NH、NRa或O；
    X选自N或CH；
    p、q彼此独立地选自0、1、2、3，且p和q不同时为0；
    r、s彼此独立地选自0、1、2、3，且r和s不同时为0；
    优选地，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团选自
    其中，Z选自CH2、CH、NH、NRa或O；
    X选自N或CH；
    r、s彼此独立地选自0、1、2、3，且r和s不同时为0；
    优选地，A选自
    其中，T选自CH2、CH、NH、NRa或O；
    Z选自CH2、CH、NH、NRa或O；
    X选自N或CH；
    n选自0、1、2、3、4或5；
    p、q彼此独立地选自0、1、2、3，且p和q不同时为0；
    r、s彼此独立地选自0、1、2、3，且r和s不同时为0；
    优选地，A选自无取代或任选被一个、两个或更多个Ra取代的稠环基团；所述稠环基团由两个、三个、四个或更多个选自苯环、吡啶环、咪唑环、哌嗪环、哌啶环、四氢吡咯环、四氢吡喃环、四氢呋喃环、呋喃环、吗啉环、环丙烷环、环丁烷环、环戊烷环、环己烷环、环庚烷环、环己亚胺环的环稠合而成；
    优选地，A选自无取代或任选被一个、两个或更多个Ra取代的
    优选地，每个Ra相同或不同，彼此独立地选自H、OH、F、Cl、氰基、甲基、乙基、异丙基、甲氧基、乙氧基、三氟甲基、二氟甲氧基、三氟甲氧基、环丙基、环丙基甲氧基；或者，连接在同一个环碳原子上的两个Ra与其连接的碳原子一起共同形成环丙烷环、环丁烷环、环戊烷环、环己烷环、环庚烷环；或者，两个不相邻的Ra以其端基相连，共同形成亚甲基或亚乙基；
    优选地，A选自
  3. 根据权利要求1或2所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物，其特征在于，E选自无取代或任选被一个或两个Re取代的下列基团：-NH-S(＝O)2-Re1、-S(＝O)2-NH-Re2、-S(＝O)(＝NH)-Re3；每个Re1、Re2、Re3相同或不同，彼此独立地选自H、C1-6烷基、羟基C1-6烷基、卤代C1-6烷基、卤代C1-6烷氧基、氰基C1-6烷基、氰基C1-6烷氧基、C3-8环烷基、3-8元杂环基、C1-6烷氧基-C1-6烷基；每个Re相同或不同，彼此独立地选自OH、卤素、氰基、C1-6烷基、C1-6烷氧基、卤代C1-6烷基、卤代C1-6烷氧基、氰基C1-6烷基、氰基C1-6烷氧基；
    优选地，E选自
  4. 根据权利要求1-3任一项所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂 化物、多晶型物、药学上可接受的盐或其前药化合物，其特征在于，M选自无取代或任选被一个、两个或更多个Rm取代的C3-8环烷基、C3-8环烯基或含氮的3-8元杂环基；每个Rm相同或不同，彼此独立地选自H、卤素、氰基、C1-6烷基、卤代C1-6烷基、氰基C1-6烷基、C1-6烷氧基、氰基C1-6烷氧基；或者，连接在同一个环碳原子上的两个Rm与其连接的碳原子一起共同形成饱和或部分不饱和的C3-8碳环；
    优选地，M选自
  5. 根据权利要求1-4任一项所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物，其特征在于，Y1、Y2、Y3不同时为N；
    优选地，当Y1为CH时，Y2为CH，Y3为N或CH；当Y1为CH时，Y2为N，Y3为N或CH；当Y1为N时，Y2为CH，Y3为N或CH；当Y1为N时，Y2为N，Y3为CH。
  6. 根据权利要求1-5任一项所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物，其特征在于，所述化合物具有以下结构：
    其中，A、M、E、Y1、Y2、Y3彼此独立地具有权利要求1-5任一项所述的定义；
    优选地，式(I)所示的化合物具有以下结构：
    其中，M、E、Y1、Y2、Y3、Ra、X、Z、T、n、p、q、r、s彼此独立地具有权利要求1-5任一项所述的定义；
    优选地，式(I)所示的化合物具有以下结构：
    其中，A、Y1、Y2、Y3独立地具有权利要求1-5任一项所述的定义；
    优选地，式(I)所示的化合物具有以下结构：
    其中，Y1、Y2、Y3、Ra、X、Z、T、n、p、q、r、s彼此独立地具有权利要求1-5任一项所述的定义；
    优选地，式(I)所示的化合物具有以下结构：
    其中，Y1、Y2、Y3、Ra、X、Z、n、r、s彼此独立地具有权利要求1-5任一项所述的定义。
  7. 根据权利要求1-6任一项所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物，其特征在于，所述化合物选自以下结构：











  8. 权利要求1-7任一项所述化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物的制备方法，包括以下步骤：
    (1)化合物a与化合物A-NH2反应得到化合物b；
    (2)化合物b与化合物E-H反应得到式(I)所示化合物；
    其中，A、E、M、Y1、Y2、Y3彼此独立地具有权利要求1-7任一项所述的定义；L选自卤素，如Cl、Br、I。
  9. 一种药物组合物，其包含治疗有效量的权利要求1-7任一项所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物中的至少一种。
  10. 权利要求1-7任一项所述的化合物、其消旋体、立体异构体、互变异构体、同位素标记物、溶剂化物、多晶型物、药学上可接受的盐或其前药化合物中的至少一种在制备药物中的用途；
    优选地，所述用途为在制备治疗KIF18A介导的病症和/或疾病的药物中的用途，如在制备KIF18A抑制剂药物中的用途；
    优选地，所述疾病例如为癌症，包括肠癌、乳腺癌、肺癌、胰腺癌、前列腺癌、膀胱癌、头颈部癌、宫颈癌或卵巢癌。
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