WO2018086592A1 - 4,5,6-三取代吲唑类衍生物、其制法与医药上的用途 - Google Patents

4,5,6-三取代吲唑类衍生物、其制法与医药上的用途 Download PDF

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WO2018086592A1
WO2018086592A1 PCT/CN2017/110465 CN2017110465W WO2018086592A1 WO 2018086592 A1 WO2018086592 A1 WO 2018086592A1 CN 2017110465 W CN2017110465 W CN 2017110465W WO 2018086592 A1 WO2018086592 A1 WO 2018086592A1
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alkyl
ring
compound
spiro
group
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PCT/CN2017/110465
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English (en)
French (fr)
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彭建彪
刘洋
王莉肖
方志华
费宗雷
陈曦
兰炯
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上海海雁医药科技有限公司
扬子江药业集团有限公司
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Priority to EP17870388.0A priority Critical patent/EP3441388A1/en
Priority to US16/305,369 priority patent/US11104664B2/en
Priority to CN201780008603.8A priority patent/CN108699029B/zh
Publication of WO2018086592A1 publication Critical patent/WO2018086592A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • 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
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings

Definitions

  • the invention belongs to the field of medical technology.
  • the invention relates in particular to a 4,5,6-trisubstituted oxazole derivative, a process for its preparation and its use as an EZH2 inhibitor, as well as pharmaceutical compositions prepared therefrom.
  • Histone-lysine-N-methyltransferase EZH2 is involved in DNA methylation and final transcriptional repression; methylation of lysine 27 by catalyzed by cofactor S-adenosyl-L-methionine To histidine H3. This methylation promotes the formation of heterochromatin, which triggers gene silencing.
  • EZH2 is a part of the functional enzyme of PRC2, which maintains the genes regulating development and differentiation through epigenetics, thus ensuring the healthy development of the embryo. Mutation or overexpression of EZH2 is associated with the formation of many cancers. EZH2 controls genes to control tumor development, and inhibition of EZH2 activity slows tumor growth. As a targeted inhibitor, EZH2 regulates a variety of cancers including breast cancer, prostate cancer, melanoma and bladder cancer.
  • the steroids WO2011140324A1 and WO2012075080A1 disclose steroids as EZH2 inhibitors for the treatment of cancer.
  • PCT application WO2012118812A2 discloses the use of bicyclic heterocyclic compounds as EZH2 inhibitors for the treatment of cancer.
  • the compounds of the invention provide a solution for disease or EZH2-mediated tumor therapy as an EZH2 inhibitor.
  • a first aspect of the invention provides a compound of formula (I), or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof:
  • R 0 is hydrogen
  • R 1 is CN, halogen (preferably fluorine, chlorine, bromine), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), halogenated C 1-8 alkyl (preferably a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl group), a C 1-8 alkoxy group (preferably a C 1-6 alkoxy group, more preferably a C 1-3) Alkoxy), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl) or C 3-8 cycloalkoxy (preferably C 3-6 cycloalkoxy);
  • C 1-8 alkyl preferably C 1-6 alkyl, more preferably C 1-3 alkyl
  • halogenated C 1-8 alkyl preferably a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl group
  • a C 1-8 alkoxy group preferably a C 1-6 al
  • R 2 is hydrogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), halogenated C 1-8 alkyl (preferably halogenated C 1-6 alkane) a group, more preferably a halogenated C 1-3 alkyl group or a C 3-8 cycloalkyl group (preferably a C 3-6 cycloalkyl group);
  • R 3 is hydrogen, halogen (preferably fluorine, chlorine, bromine), hydroxyl group, CN, C 1-8 alkyl group (preferably C 1-6 alkyl group, more preferably C 1-3 alkyl group), halogenated C 1 -8 alkyl (preferably halogenated C 1-6 alkyl, more preferably halogenated C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), C 1 A-8 alkoxy group (preferably a C 1-6 alkoxy group, more preferably a C 1-3 alkoxy group), a halogenated C 1-8 alkoxy group (preferably a halogenated C 1-6 alkoxy group, More preferably, it is a halogenated C 1-3 alkoxy group, a C 3-8 cycloalkoxy group (preferably a C 3-6 cycloalkoxy group), a C 6-10 aryl group (preferably a phenyl group), a -C ( O) C 1-8 al
  • X is NR 4 , CR 5 R 6 , O, S or S(O) 2 ;
  • R 4 is hydrogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), halogenated C 1-8 alkyl (preferably halogenated C 1-6 alkane) More preferably, it is a halogenated C 1-3 alkyl group, C(O)C 1-8 alkyl group (preferably C(O)C 1-6 alkyl group, more preferably C(O)C 1-3 Alkyl), C(O)OC 1-8 alkyl (preferably C(O)OC 1-6 alkyl, more preferably C(O)OC 1-3 alkyl), CONR a1 R b1 , C 3 -8 cycloalkyl (preferably C 3-6 cycloalkyl), -SO 2 C 1-10 alkyl (preferably -SO 2 C 1-6 alkyl, more preferably -SO 2 C 1-3 alkane Base, -C(O)CH 2 CN, -C(O)CH 2 OH, 4 to 6-membered saturated monoheterocycle
  • R 5 and R 6 are each independently hydrogen, halogen, hydroxy, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), halogenated C 1-8 alkyl (preferably a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl group), a C 3-8 cycloalkyl group (preferably a C 3-6 cycloalkyl group), NR a2 R b2 , 4 a 6-membered saturated monoheterocyclic ring, a 5- to 6-membered monocyclic heteroaryl ring, a spiro ring, a spiro heterocyclic ring, a bridged ring or a bridged heterocyclic ring;
  • R a , R b , R a1 , R b1 , R a2 , and R b2 are each independently hydrogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), C(O)C 1-8 alkyl (preferably C(O)C 1-6 alkyl, more preferably C(O)C 1-3 alkyl);
  • the A ring is a structure represented by the formula (A-1), the formula (A-2), the formula (A-3) or the formula (A-4):
  • R 11 , R 21 , R 31 and R 41 are each independently hydrogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), halogen C 1 -8 alkyl (preferably halogenated C 1-6 alkyl, more preferably halogenated C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), 4 to a 6-membered saturated monoheterocyclic ring, a 5- to 6-membered monocyclic heteroaryl ring, a spiro ring, a spiro heterocyclic ring, a bridged ring or a bridged heterocyclic ring;
  • R 12 , R 22 , R 32 and R 42 are each independently hydrogen, halogen (preferably fluorine, chlorine, bromine), C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 Alkyl), halogenated C 1-8 alkyl (preferably halogenated C 1-6 alkyl, more preferably halogenated C 1-3 alkyl) or C 3-8 cycloalkyl (preferably C 3- 6 cycloalkyl);
  • halogen preferably fluorine, chlorine, bromine
  • C 1-8 alkyl preferably C 1-6 alkyl, more preferably C 1-3 Alkyl
  • halogenated C 1-8 alkyl preferably halogenated C 1-6 alkyl, more preferably halogenated C 1-3 alkyl
  • C 3-8 cycloalkyl preferably C 3- 6 cycloalkyl
  • the bridged or bridged heterocyclic ring is unsubstituted or substituted with 1, 2 or 3 substituents selected from the group consisting of CN, acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, -C(O) OC 1-6 alkyl, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), C 1-8 alkoxy (preferably C 1-6 alkoxy) More preferably, it is a C 1-3 alkoxy group, a halogenated C 1-8 alkyl group (preferably a halogenated C 1-6 alkyl group, more preferably a
  • R 1 is C 1-3 alkyl (preferably methyl or ethyl).
  • R 2 is hydrogen, C 1-3 alkyl or halo C 1-3 alkyl.
  • R 2 is C 1-3 alkyl (preferably methyl or ethyl).
  • R 3 is hydrogen, halogen, C 1-3 alkyl or C 1-3 alkoxy.
  • R 3 is hydrogen, fluorine, chlorine, methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, propoxy or isopropoxy.
  • R 0 is hydrogen and R 1 is C 1-3 alkyl (preferably methyl or ethyl).
  • R 11 , R 21 , R 31 , R 41 are each independently hydrogen or -(CH 2 ) n -L 1 ; wherein L 1 is CN, NR a3 R b3 , C 1-8 alkane a group (preferably a C 1-6 alkyl group, more preferably a C 1-3 alkyl group), a C 1-8 alkoxy group (preferably a C 1-6 alkoxy group, more preferably a C 1-3 alkoxy group) a halogenated C 1-8 alkyl group (preferably a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl group), a C 3-8 cycloalkyl group (preferably a C 3-6 ring)
  • the alkyl group, alkoxy group, cycloalkyl group, 4 to 6 membered saturated monoheterocyclic ring, 5 to 6 membered monocyclic heteroaryl ring, spiro ring, spiro heterocyclic ring, bridged ring or bridged heterocyclic ring is unsubstituted Or substituted with one substituent selected from the group consisting of acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, -C(O)OC 1-6 alkyl, C 1-3 alkyl, C 1 -3 alkoxy, halo C 1-3 alkyl, C 3-6 cycloalkyl, NR a3 R b3 , azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, Piperidine, oxazolidine, piperazine, dioxolane, dioxane,
  • n is 0 or 1.
  • the 4- to 6-membered saturated monoheterocyclic ring is azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, sulfur Demorpholine, thiomorpholine-1,1-dioxide or tetrahydropyran.
  • the 5- to 6-membered monocyclic heteroaryl ring is thiophene, N-alkylpyrrole, furan, thiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, tetrazole, metabolite An oxazole, an oxadiazole, a thiadiazole, a pyridine, a pyridazine, a pyrimidine or a pyrazine.
  • R 11 , R 21 , R 31 , and R 41 are each independently hydrogen, methyl, ethyl, n-propyl, isopropyl, cyclopropyl, monofluoromethyl, difluoromethyl.
  • Base trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl.
  • each of R 12 , R 22 , R 32 and R 42 is independently hydrogen, methyl, ethyl, n-propyl or isopropyl.
  • R 12 , R 22 , R 32 , and R 42 are each independently hydrogen.
  • R 4 is hydrogen or -(CH 2 ) m -L 2 ; wherein L 2 is CN, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1- 3 alkyl), NR a3 R b3 , C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy), halogenated C 1-8 alkyl (preferred) It is a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl group, or a C(O)C 1-8 alkyl group (preferably a C(O)C 1-6 alkyl group, more preferably C(O)C 1-3 alkyl), C(O)OC 1-8 alkyl (preferably C(O)OC 1-6 alkyl, more preferably C(O)OC 1-3 alkyl) , CONR a1 R b1 , C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl (preferably C
  • the bridged or bridged heterocyclic ring is unsubstituted or substituted with one substituent selected from the group consisting of acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxy, -C(O)OC 1-6 alkyl , C 1-3 alkyl, C 1-3 alkoxy, halo C 1-3 alkyl, C 3-6 cycloalkyl, NR a3 R b3 , azetidine, oxetane, Tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, oxazolidine
  • m is 0 or 1.
  • R 4 is -(CH 2 ) m -L 2 ; wherein L 2 is a halogenated C 1-8 alkyl group (preferably a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl) or 4 to 6-membered saturated monoheterocycle; m is 0 or 1.
  • L 2 is a halogenated C 1-8 alkyl group (preferably a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl) or 4 to 6-membered saturated monoheterocycle; m is 0 or 1.
  • R 4 is -CH 2 -L 2 ; wherein L 2 is a halogenated C 1-8 alkyl group (preferably a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl group) alkyl).
  • R 4 is L 2 ; wherein L 2 is a 4 to 6 membered saturated monoheterocyclic ring.
  • the 4- to 6-membered saturated monoheterocyclic ring is azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, sulfur Demorpholine, thiomorpholine-1,1-dioxide or tetrahydropyran.
  • the 5- to 6-membered monocyclic heteroaryl ring is thiophene, N-alkylpyrrole, furan, thiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, tetrazole, metabolite An oxazole, an oxadiazole, a thiadiazole, a pyridine, a pyridazine, a pyrimidine or a pyrazine.
  • R 4 is hydrogen, halogenated C 1-3 alkyl.
  • R 5 is hydrogen and R 6 is -(CH 2 ) p -L 3 ;
  • L 3 is CN, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3 alkyl), C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy), halogenated C 1-8 alkyl (preferably halogenated) C 1-6 alkyl, more preferably halogenated C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), NR a3 R b3 , 4 to 6-member saturated single a heterocyclic ring, a 5- to 6-membered monocyclic heteroaryl ring, a spiro ring, a spiro heterocyclic ring, a bridged ring or a bridged heterocyclic ring; p is 0, 1 or 2;
  • the alkyl group, alkoxy group, cycloalkyl group, 4 to 6 membered saturated monoheterocyclic ring, 5 to 6 membered monocyclic heteroaryl ring, spiro ring, spiro heterocyclic ring, bridged ring or bridged heterocyclic ring is unsubstituted Or substituted with one substituent selected from the group consisting of acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, -C(O)OC 1-6 alkyl, C 1-3 alkyl, C 1 -3 alkoxy, halo C 1-3 alkyl, C 3-6 cycloalkyl, NR a3 R b3 , azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, Piperidine, oxazolidine, piperazine, dioxolane, dioxane,
  • p is 0, 1 or 2;
  • L 3 is NR a3 R b3 or a 4 to 6-membered saturated monoheterocyclic ring;
  • R a3 and R b3 are each independently hydrogen, C 1-3 alkyl or a C 1-3 alkoxy-substituted C 1-3 alkyl group; the alkyl group, alkoxy group or a 4 to 6 membered saturated monoheterocyclic ring is unsubstituted or substituted with one substituent selected from the group consisting of : acetyl, hydroxy, hydroxymethyl, hydroxyethyl, carboxyl, -C(O)OCH 3 , -C(O)OCH 2 CH 3 , -C(O)OC(CH 3 ) 3 , -C(O OCH(CH 3 ) 2 , methyl, ethyl, n-propyl, isopropyl, methoxy, ethoxy, propoxy, isopropoxy,
  • the 4- to 6-membered saturated monoheterocyclic ring is azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, piperazine, morpholine, sulfur Demorpholine, thiomorpholine-1,1-dioxide or tetrahydropyran.
  • the 5- to 6-membered monocyclic heteroaryl ring is thiophene, N-alkylpyrrole, furan, thiazole, imidazole, oxazole, pyrrole, pyrazole, triazole, tetrazole, metabolite An oxazole, an oxadiazole, a thiadiazole, a pyridine, a pyridazine, a pyrimidine or a pyrazine.
  • R 11 , R 12 , R 21 , R 22 , R 31 , R 32 , R 41 and R 42 are as defined above.
  • the compound is selected from the following Table A:
  • the compound is selected from the following Table B:
  • a second aspect of the invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising the compound of the first aspect of the invention, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof; A pharmaceutically acceptable carrier.
  • a third aspect of the invention provides a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, or a pharmaceutical composition according to the second aspect of the invention In the preparation of EZH2 inhibitors.
  • a third aspect of the invention provides a compound according to the first aspect of the invention, or a pharmaceutically acceptable salt, solvate, stereoisomer or prodrug thereof, or a pharmaceutical composition according to the second aspect of the invention Use in the preparation of a disease or condition mediated by EZH2.
  • a fourth aspect of the invention provides a method of treating a disease or condition mediated by EZH2 comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or a solvent thereof A compound, a stereoisomer or a prodrug, or a pharmaceutical composition according to the second aspect of the invention.
  • a fifth aspect of the invention provides a method of treating a disease or condition mediated by EZH2 comprising administering to a subject in need thereof a therapeutically effective amount of a compound of the first aspect of the invention, or a pharmaceutically acceptable salt thereof, or a solvent thereof a compound, a stereoisomer or a prodrug, and another therapeutically active agent.
  • the disease or condition mediated by EZH2 is selected from the group consisting of cancer, pulmonary hypertension, myelofibrosis, human immunodeficiency virus (HIV) disease, graft versus host disease (GVHD), Weaver syndrome, Psoriasis vulgaris or liver fibrosis.
  • HIV human immunodeficiency virus
  • GVHD graft versus host disease
  • Weaver syndrome Psoriasis vulgaris or liver fibrosis.
  • the disease or condition mediated by EZH2 is cancer.
  • the cancer mediated by EZH2 includes, but is not limited to, thyroid cancer, cardiac sarcoma, lung cancer, gastrointestinal cancer, genitourinary tract tumor, liver cancer, mantle cell lymphoma, osteosarcoma, nervous system sarcoma, Gynecological cancer, hematological tumor, adrenal neuroblastoma, skin cancer, astrocytic tumor, breast cancer, colorectal cancer, endometrial cancer, head and neck cancer, oral cancer.
  • the inventors have unexpectedly discovered such 4,5,6-trisubstituted oxazole derivatives, especially the 7-substituted unsubstituted 4,5,6-trisubstituted oxazoles at the 7-position of carbazole.
  • the derivative has high inhibitory activity against enzymes such as EZH2Y641F and SU-DHL-6 and SU-DHL-10. Therefore, this series of compounds is expected to be developed as a drug for treating tumors. On this basis, the inventors completed the present invention.
  • alkyl refers to both straight and branched saturated aliphatic hydrocarbon groups, and C1-8 alkyl is alkyl having from 1 to 8 carbon atoms, preferably C1-6 alkyl, more preferably C 1-3 alkyl, the definition is similar; non-limiting examples of alkyl include: methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl , n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3 -methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl Butyl, 2,2-dimethylbutyl, 1,3-dimethylbut
  • cycloalkyl refers to a saturated or partially unsaturated monocyclic cyclic hydrocarbon group
  • C 3-8 cycloalkyl refers to a cyclic hydrocarbon group containing from 3 to 8 carbon atoms, preferably a C 3-6 ring.
  • Non-limiting examples of cycloalkyl groups include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatrienyl
  • a cyclooctyl group or the like is preferably a cyclopropyl group, a cyclopentyl group or a cyclohexenyl group.
  • spirocyclic refers to a polycyclic group that shares a carbon atom (called a spiro atom) between the individual rings, which may contain one or more double bonds, but none of the rings have fully conjugated ⁇ electrons. system.
  • the spiro ring is divided into a double spiro ring or a multi-spiral ring depending on the number of rings, preferably a double spiro ring. More preferably, it is preferably a 4 member/5 member, a 5 member/5 member or a 5 member/6 member double screw ring.
  • spiroheterocycle refers to a polycyclic hydrocarbon in which one atom (called a spiro atom) is shared between monocyclic rings, wherein one or two ring atoms are selected from nitrogen, oxygen or S(O) n (where n is an integer) From 0 to 2), the remaining atoms are carbon. These may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system.
  • the spiroheterocycle is classified into a bispiral heterocyclic ring or a polyspirocyclic ring according to the number of rings, preferably a double spiro heterocyclic ring. More preferably, it is 4 yuan/5 yuan, 5 yuan/5 yuan or 5 yuan / 6 yuan double spiro heterocycle.
  • bridged ring refers to a polycyclic group that shares two or more carbon atoms.
  • the shared carbon atom is called the bridgehead carbon.
  • the two bridgehead carbons may be carbon chains or a bond. , called the bridge. These may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system. It is preferably a bicyclic or tricyclic bridged ring.
  • bridge heterocycle refers to a polycyclic group that shares two or more atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or S(O) n (where n is an integer from 0 to 2) a hetero atom, the remaining ring atoms being carbon. These may contain one or more double bonds, but none of the rings have a fully conjugated pi-electron system. It is preferably a bicyclic or tricyclic bridge heterocycle. E.g:
  • 8- to 10-membered bicyclic refers to a bridged ring containing two rings of 8 to 10 ring atoms, which may be a saturated all-carbon bicyclic or partially unsaturated all-carbon bicyclic ring, examples of which include ( But not limited to):
  • 8- to 10-membered bicyclic heterocycle refers to a two-ring-containing bridged heterocyclic ring containing from 8 to 10 ring atoms, wherein 1, 2, 3, 4 or 5 ring carbon atoms are selected from nitrogen Substituted by a hetero atom of oxygen or sulfur.
  • bicyclic heterocycles include, but are not limited to, tetrahydroquinoline rings, tetrahydroisoquinoline rings, decahydroquinoline rings, and the like.
  • C 1-8 alkoxy refers to -O-(C 1-8 alkyl), wherein alkyl is as defined above.
  • a C 1-6 alkoxy group is preferred, and a C 1-3 alkoxy group is more preferred.
  • Non-limiting examples include methoxy, ethoxy, propoxy, isopropoxy, butoxy, tert-butoxy, isobutoxy, pentyloxy and the like.
  • C 3-8 cycloalkoxy refers to -O-(C 3-8 cycloalkyl), wherein cycloalkyl is as defined above. Preference is given to C 3-6 cycloalkoxy. Non-limiting examples include cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy, and the like.
  • C 6-10 aryl refers to an all-carbon monocyclic or fused polycyclic ring (ie, a ring that shares a pair of adjacent carbon atoms) having a conjugated ⁇ -electron system, meaning 6 to 10
  • An aryl group of a carbon atom preferably a phenyl group and a naphthyl group, more preferably a phenyl group.
  • a bond refers to the attachment of two groups attached thereto through a covalent bond.
  • halogen refers to fluoro, chloro, bromo or iodo.
  • halo means that one or more (eg 1, 2, 3, 4 or 5) hydrogens in the group are replaced by a halogen.
  • halo C 1-8 alkyl refers to an alkyl group substituted with one or more (eg 1, 2, 3, 4 or 5) halo, wherein alkyl is as defined above. It is selected as a halogenated C 1-6 alkyl group, more preferably a halogenated C 1-3 alkyl group.
  • halogenated C 1-8 alkyl groups include, but are not limited to, monochloromethyl, dichloromethyl, trichloromethyl, monochloroethyl, 1,2-dichloroethyl, trichloroethyl, Monobromoethyl, monofluoromethyl, difluoromethyl, trifluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, and the like.
  • halo C 1-8 alkoxy means that the alkoxy group is substituted by one or more (eg 1, 2, 3, 4 or 5) halogens, wherein the alkoxy group is as defined above. It is preferably a halogenated C 1-6 alkoxy group, more preferably a halogenated C 1-3 alkoxy group. These include, but are not limited to, trifluoromethoxy, trifluoroethoxy, monofluoromethoxy, monofluoroethoxy, difluoromethoxy, difluoroethoxy, and the like.
  • halo C 3-8 cycloalkyl refers to a cycloalkyl group substituted with one or more (eg, 1, 2, 3, 4, or 5) halo, wherein cycloalkyl is as defined above.
  • Preferred is a halogenated C 3-6 cycloalkyl group. These include, but are not limited to, trifluorocyclopropyl, monofluorocyclopropyl, monofluorocyclohexyl, difluorocyclopropyl, difluorocyclohexyl, and the like.
  • deuterated C 1-8 alkyl refers to an alkyl group substituted with one or more (eg 1, 2, 3, 4 or 5) deuterium atoms, wherein alkyl is as defined above. It is preferably a deuterated C 1-6 alkyl group, more preferably a deuterated C 1-3 alkyl group. Examples of deuterated C 1-20 alkyl groups include, but are not limited to, monodeuterated methyl, monodeuterated ethyl, dideuterated methyl, didecanoethyl, triterpene methyl, triterpenoid Base.
  • amino refers to NH 2
  • cyano refers to the CN
  • Niro refers to NO 2
  • benzyl refers to -CH 2 - phenyl
  • carboxy refers to -C (O) OH
  • acetyl means a -C (O) CH 3
  • hydroxymethyl group refers to -CH 2 OH
  • hydroxyethyl refers to -CH 2 CH 2 OH
  • hydroxy means - OH
  • thiol refers to SH
  • cyclopropylene structure is:
  • heteroaryl ring and “heteroaryl” are used interchangeably and mean having 5 to 10 ring atoms, preferably 5 or 6 membered monocyclic heteroaryl or 8 to 10 membered bicyclic heteroaryl.
  • the ring array shares 6, 10 or 14 ⁇ electrons; and has a group of 1 to 5 hetero atoms in addition to carbon atoms.
  • Hetero atom means nitrogen, oxygen or sulfur.
  • 3- to 6-membered saturated or partially unsaturated monocyclic refers to a saturated or partially unsaturated, all-carbon monocyclic ring containing from 3 to 6 ring atoms.
  • monocyclic rings include, but are not limited to, cyclopropyl rings, cyclobutyl rings, cyclopentyl rings, cyclopentenyl rings, cyclohexyl rings, cyclohexenyl rings, cyclohexadienyl rings, cycloheptyl groups. Ring, cycloheptatrienyl ring, cyclooctyl ring, and the like.
  • 3 to 6 membered saturated monoheterocycle means that 1, 2 or 3 carbon atoms in a 3 to 6 membered monocyclic ring are selected from nitrogen, oxygen or S(O) t (where t is an integer 0)
  • the heteroatoms to 2) are substituted, but do not include the ring moiety of -OO-, -OS- or -SS-, and the remaining ring atoms are carbon; preferably 4 to 6 members, more preferably 5 to 6 members.
  • saturated monoheterocycles include, but are not limited to, propylene oxide, azetidine, oxetane, tetrahydrofuran, tetrahydrothiophene, tetrahydropyrrole, piperidine, pyrroline, oxazolidine, piperazine , dioxolane, dioxane, morpholine, thiomorpholine, thiomorpholine-1,1-dioxide, tetrahydropyran, and the like.
  • a "5- to 6-membered monocyclic heteroaryl ring” refers to a monoheteroaryl ring containing from 5 to 6 ring atoms, including, for example, but not limited to, a thiophene ring, an N-alkylpyrrole ring, Furan ring, thiazole ring, imidazole ring, oxazole ring, pyrrole ring, pyrazole ring, triazole ring, tetrazole ring, isoxazole ring, oxadiazole ring, thiadiazole ring, pyridine ring, pyridazine ring, Pyrimidine ring, pyrazine ring and the like.
  • 8- to 10-membered bicyclic heteroaryl ring refers to a bi-heteroaryl ring containing from 8 to 10 ring atoms, including, for example, but not limited to, benzofuran, benzothiophene, anthracene, Isoindole, quinoline, isoquinoline, carbazole, benzothiazole, benzimidazole, quinazoline, quinoxaline, porphyrin, pyridazine.
  • substituted refers to one or more hydrogen atoms in the group, preferably 1 to 5 hydrogen atoms are independently substituted with each other by a corresponding number of substituents, more preferably 1 to 3 hydrogen atoms are independent of each other. The ground is replaced by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino group or a hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.
  • any of the groups herein may be substituted or unsubstituted.
  • the substituent is preferably a group of 1 to 5 or less, independently selected from the group consisting of CN, halogen, C 1-8 alkyl (preferably C 1-6 alkyl, more preferably C 1-3) Alkyl), C 1-8 alkoxy (preferably C 1-6 alkoxy, more preferably C 1-3 alkoxy), halogenated C 1-8 alkyl (preferably halogen C 1- 6 alkyl, more preferably halogenated C 1-3 alkyl), C 3-8 cycloalkyl (preferably C 3-6 cycloalkyl), halogenated C 1-8 alkoxy (preferably halogenated) C 1-6 alkoxy, more preferably halogenated C 1-3 alkoxy), C 1-8 alkyl substituted amine, amine, halogenated C 1-8 alkyl substituted amine, 4 Up to 6-membered saturated monoheterocyclic ring, 5-
  • EZH2 inhibitor refers to an agent (in the present invention, a compound of formula (I)) capable of inhibiting the increase in the expression of histone lysine N-methyltransferase EZH2, which is PRC2
  • the catalytic functional subunit is responsible for the Lys27 methylation of a specific histone H3 (H3K27) and is indispensable for stem cell self-renewal.
  • a disease or condition mediated by EZH2 refers to an abnormal episode formed in a patient due to abnormal expression of the histone lysine N-methyltransferase EZH2 resulting in an abnormal epigenetic modification.
  • terapéuticaally effective amount refers to a compound of the invention that will elicit a biological or medical response to an individual, such as reducing or inhibiting the activity of an enzyme or protein or ameliorating a condition, alleviating a condition, slowing or delaying the progression of a disease, or preventing a disease, and the like. the amount.
  • pharmaceutically acceptable carrier means a non-toxic, inert, solid, semi-solid substance or liquid filler, diluent, encapsulating or auxiliary formulation or any type of excipient that is compatible with the patient, most Preferably, it is a mammal, more preferably a human, which is suitable for delivering the active agent to a target of interest without terminating the activity of the agent.
  • patient refers to an animal, preferably a mammal, and more preferably a human.
  • mammal refers to warm-blooded vertebrate mammals including, for example, cats, dogs, rabbits, bears, foxes, wolves, monkeys, deer, rats, pigs, and humans.
  • treating refers to alleviating, delaying progression, attenuating, preventing, or maintaining an existing disease or condition (eg, cancer). Treatment also includes curing, preventing, or alleviating one or more symptoms of the disease or condition to some extent.
  • the present invention provides a process for the preparation of a compound of formula (I), which can be prepared by a variety of synthetic procedures. Exemplary methods of preparation of these compounds can include, but are not limited to, the procedures described below.
  • the compounds of formula (I) of the present invention can be prepared by the following schemes and exemplary methods described in the Examples and related publications used by those skilled in the art.
  • the steps in the method can be extended or merged as needed during the specific operation.
  • Step 1 The nitro compound is converted to the corresponding amine compound, which can be reduced under acidic conditions with a metal (may be, but not limited to, iron powder, zinc powder); or hydrogenated under palladium on carbon catalysis.
  • a metal may be, but not limited to, iron powder, zinc powder
  • Step 2 The amine and ketone at the 4-position of the carbazole compound can be subjected to nucleophilic addition via a catalyst (which may be, but is not limited to, sodium triacetoxyborohydride) under acidic conditions.
  • a catalyst which may be, but is not limited to, sodium triacetoxyborohydride
  • Step 3 The nucleophilic addition reaction conditions of the amine and the aldehyde are the same as in the step 2.
  • Step 4 the bromine reaction on the aromatic ring of the carbazole compound and the carbonylation reaction of, for example, hexacarbonyl molybdenum under the action of a palladium catalyst, and then condensing with an amine to obtain the target compound (I);
  • the palladium catalyst used may be, but not limited to, Pd. (dppf)Cl 2
  • the base used may be, but not limited to, triethylamine.
  • Step 1 The nitro compound is converted to the corresponding amine compound, which can be reduced under acidic conditions with a metal (may be, but not limited to, iron powder, zinc powder); or hydrogenated under palladium on carbon catalysis.
  • a metal may be, but not limited to, iron powder, zinc powder
  • Step 2 The amine and ketone at the 4-position of the carbazole compound can be subjected to nucleophilic addition via a catalyst (which may be, but is not limited to, sodium triacetoxyborohydride) under acidic conditions.
  • a catalyst which may be, but is not limited to, sodium triacetoxyborohydride
  • Step 3 The nucleophilic addition reaction conditions of the amine and the aldehyde are the same as in the step 2.
  • Step 4 Hydrolysis of the ester, the compound may be hydrolyzed in an alkaline solution such as methanol, and the base used may be, but not limited to, sodium hydroxide.
  • Step 5 The acid of the carbazole compound is condensed with an amine to form an amide compound (I) under the action of a condensing agent.
  • a series of novel 4,5,6-trisubstituted carbazole derivatives are provided, which have high inhibitory activity against EZH2 and can be used as a medicament for treating tumors.
  • DMB is 2,4-dimethoxybenzyl
  • THF is tetrahydrofuran
  • EA is ethyl acetate
  • PE is petroleum ether
  • Ac 2 O is acetic anhydride
  • NBS is N-bromosuccinimide.
  • DCM is dichloromethane
  • AIBN is azobisisobutyronitrile
  • Pd(dppf)Cl 2 is 1,1'-bis(diphenylphosphino)ferrocene]palladium dichloride
  • TFA is trifluoroacetic acid
  • TBSCl Is tert-butyldimethylchlorosilane
  • NCS is N-chlorosuccinimide
  • DHP is dihydropyran
  • LiAlH 4 is lithium aluminum hydride
  • PMB is p-methoxybenzyl
  • LiHMDS is two (three Methylsilyl) lithium amide
  • Pd 2 (dba) 3 is tris(dibenzylideneacetone)dipalladium
  • RuPhos is 2-dicyclohexylphosphorin-2',6'-diisopropoxy-1,1 '-Biphenyl
  • DMAP is 4-dimethylaminopyridine
  • room temperature means about 20-25 °C.
  • Step 1 A solution of the compound malononitrile (12 g, 181.7 mmol) in dry tetrahydrofuran (225 mL) was stirred for 1 hour in ice-cooling, and then sodium hydrogen (4.8 g, 199.8 mmol) was added portionwise and stirred for 2 hours, and compound 2a was added dropwise. -1 (16.8 g, 199.8 mmol), the mixture was slowly warmed to room temperature for 1 hour. The reaction mixture was quenched with aq. EtOAc (EtOAc)EtOAc. MS m/z (ESI): 151 [M+H] + .
  • Step 2 A mixture of compound 2a-2 (28 g, 181.7 mmol), EtOAc (2. The reaction solution was filtered, and the solid residue was crystallized from methanol to yield 25 g of Compound 2a-3. MS m/z (ESI): 151 [M+H] + .
  • Step 3 Compound 2a-3 (80 mg, 0.53 mmol) was added 2 mL of phosphorus oxychloride, and the mixture was stirred at 100 ° C for 2 hours. LC-MS was followed until the reaction was complete. The mixture was cooled, poured into ice water, adjusted to pH 8 with aqueous ammonia, extracted with ethyl acetate and then dried and evaporated. MS m/z (ESI): 169 [M+H] + .
  • Step 4 A mixture of compound 2a-4 (300 mg, 1.78 mmol), sodium methoxide (481 mg, 8.9 mmol) and methanol (15 mL). LC-MS was followed until the reaction was complete. The solvent was removed by concentration. MS m/z (ESI): 495 [M+H] + .
  • Step 5 The preparation method is the same as Compound 1a except that Compound 1a-1 in Process 1a is replaced by Compound 2a-5. MS m/z (ESI): 152 [M+H] + .
  • Step 1 A solution of compound 3a-1 (1 g, 4.35 mmol) in EtOAc (10 mL) EtOAc (EtOAc (EtOAc) .
  • EtOAc EtOAc
  • LC-MS was followed until the reaction was complete.
  • the reaction liquid was filtered, and the residue obtained by concentration of the filtrate was dissolved in water, and then filtered to give a white solid compound 3a-2.
  • Step 2 A solution of compound 3a-2 (500 mg, 45 mmol) in EtOAc (EtOAc) (EtOAc) LC-MS was followed until the reaction was complete. The mixture was poured into ice water, the pH was adjusted to 8, and the precipitated solid was filtered to give 400 mg of Compound 3a-3. MS m/z (ESI): 247 [M+H] + .
  • Step 4 The preparation method was the same as the compound 3a-2 except that the compound 3a-1 in the 3a-2 process was replaced by the compound 3a-4, and purified by combiflash to obtain a red solid compound 3a (800 mg, 45%). MS m/z (ESI): 226 [M+H] + .
  • Step 1 Compound 3a (800 mg, 3.54 mmol), tetrahydropyranone (708 mg, 7.08 mmol), and trifluoroacetic acid 5 mL 1,4-dioxane (50 mL) was stirred at room temperature for 2 hr. Sodium oxyborohydride (2.25 mg, 10.62 mmol) was stirred at room temperature for 4 hours. LC-MS was followed until the reaction was complete. The reaction solution was poured into water, and the mixture was adjusted to pH 8 with sodium hydrogen carbonate solution, extracted with ethyl acetate, dried and concentrated to yield 1 g of Compound 4a-1. MS m/z (ESI): 312.2 [M+H] + .
  • Step 2 Compound 4a-1 (1 g, 3.22 mmol), acetaldehyde (710 mg, 16.12 mmol), and 3 mL of THF in 1,4-dioxane (30 mL). Sodium hydride (2.05 g, 9.67 mmol) was stirred at room temperature overnight. LC-MS was followed until the reaction was complete. The reaction solution was poured into water, and the mixture was adjusted to pH 8 with sodium hydrogen carbonate solution, and ethyl acetate was evaporated and concentrated, and purified by combiflash to give red compound 4a (280 mg, 26%). MS m/z (ESI): 340.1 [M+H] + .
  • Step 1 To a solution of compound 5a-1 (8 g, 35.3 mmol) in EtOAc (EtOAc) After the reaction mixture was cooled, it was concentrated to remove most of the solvent, filtered, and the filter cake was dried under reduced pressure to give 7.5 g of Compound 5a-2. MS m/z (ESI): 241 [M+H]+.
  • Step 2 Compound 5a-2 (7.6 g, 31.64 mmol) in acetic acid (150 mL) was added portionwise to iron powder (14 g, 253 mmol), and the reaction temperature was controlled at 20-30 ° C. After the addition, stirring was continued for 10 min, LC- The MS tracks until the end of the reaction. The reaction mixture was filtered, and the filtrate was evaporated,jjjjjjjjjj MS m/z (ESI): 211 [M+H]+.
  • Step 4 The preparation method is the same as the compound 3a-4 except that the compound 3a-3 in the 3a-4 process is replaced with the compound 5a-4-1.
  • Step 5 Compound 5a-45 (3 g, 0.01 mol), methylene iodide (4.2 g, 0.03 mol), potassium carbonate (2.2 g, 0.015 mol) in DMF was stirred at room temperature for 2 h, and LC-MS was followed until the end of the reaction. The reaction mixture was poured into water (150 ml), stirred for 30 min, filtered, and then filtered and evaporated to dryness to give Compound 5a-6 (2.7 g, 86%), MS m/z (ESI): 316.1 [M+H]+.
  • Step 6 The preparation method is the same as the compound 3a-4 except that the compound 3a-3 in the 3a-4 process is replaced with the compound 5a-6.
  • Step 67 Compound 5a-67 (8 g, 0.026 mol), potassium trifluorovinylborate (7 g, 0.052 mol), Pd(dppf)Cl2 (0.95 g, 0.0013 mol), sodium carbonate (5.5 g, 0.052 mol)
  • the mixed solution of dioxane/water (100 ml/10 ml) was stirred at 100 ° C overnight, and LC-MS was followed until the end of the reaction. After the reaction mixture was cooled to room temperature, the mixture was filtered, evaporated, evaporated, mjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj m/z (ESI): 2632.2 [M+H]+.
  • Step 7 Compound 5a-78 (1 g, 4.4 mmol), EtOAc (EtOAc) The reaction mixture was filtered, EtOAcjjjjjjjjjj
  • Step 1 To a solution of Compound 6a-1 (2 g, EtOAc (EtOAc) (EtOAc) . The reaction mixture was poured into water and extracted with methylene chloride. 60%), MS m/z (ESI): 257.0 [M+H]+.
  • Step 2 Compound 5a-5 (2 g, 6.67 mmol), 6a-2 (2.56 g, 10 mmol), potassium carbonate (1.84 g, 13.33 mmol), and a solution of potassium iodide (1.66 g, 10 mmol) in acetone for 8 h. After the reaction mixture was cooled to room temperature, filtered, EtOAc EtOAcjjjjjjj
  • Step 1 Compound 5a-5 (20 g, 0.067 mol), NIS (16.6 g, 0.074 mol) in DMF was stirred at 60 ° C for 3 h and LC-MS was followed until the end of the reaction. After the reaction mixture was cooled to room temperature, poured into water (600 ml), stirred for 30 min, filtered, filtered, washed with water and dried to give compound 10a-1 (25 g), MS m/z (ESI): 425.7 [M+H]+.
  • Step 2 Compound 10a-1 (25 g, 0.059 mol), potassium carbonate (12.3 g, 0.09 mol), iodomethane (12.8 g, 0.09 mol) in DMF (100 ml) .
  • the reaction mixture was poured into water, filtered, and then filtered.
  • Step 3 To compound 10a-2 (400 mg, 0.91 mmol) and trimethylcyclotriboroxane (568 mg, 4.55 mmol), potassium carbonate (376 mg, 2.73 mmol) in dioxane/water (5 ml / 1 ml) A catalytic amount of Pd(dppf)Cl2 was added to the mixed solution, and the reaction was stirred at 105 ° C for 16 hours under nitrogen atmosphere, and LC-MS was traced until the end of the reaction. Water was added to the reaction mixture, and the mixture was evaporated. EtOAcjjjjjjjj :264[M+H]+.
  • Step 4 Same as the preparation of Step 2 in Intermediate 7a, MS m/z (ESI): 234 [M+H]+.
  • Step 1 Slowly add zinc powder to a mixed solution of compound 10a-1 (16.4 g, 37.3 mmol) in methanol/glacial acetic acid (400 ml/400 ml) at -5 to 0 ° C, and stir at 0 ° C for 20 min. Then stir at room temperature for another 20 min, LC-MS Track to the end of the reaction. The reaction solution was filtered through celite, and water (1L) was applied to the filtrate, and the pH was adjusted to 7-8 with a saturated sodium carbonate solution. The mixture was extracted with methylene chloride. EtOAc (EtOAc m.) M+H]+.
  • Step 2 Compound 11a-1 (300 mg, 1.06 mmol), zinc cyanide (250 mg, 2.21 mmol), tetratriphenylphosphine palladium (128 mg, 0.11 mmol) in DMF (4 mL) The microwave reaction was carried out for 30 min, and LC-MS was traced until the end of the reaction. Water and ethyl acetate were added to the reaction mixture, and the organic layer was evaporated. The crude product was recrystallized from EtOAc (EtOAc:EtOAc)
  • Step 1 Refer to the preparation method of 5a-7 except that potassium trifluorovinylborate was replaced with methylboronic acid, and sodium carbonate was replaced with cesium carbonate. MS m/z (ESI): 250.3 [M+H]+.
  • Step 2 To a solution of the compound 12a-1 (672 mg, 2.7 mmol) in acetic acid (20 ml), EtOAc (2. The reaction mixture was filtered, and then evaporated, mjjjjjjjjj
  • Step 3 To a solution of Compound 12a-2 (700 mg, 2.62 mmol) in MeOH (25 ml), The reaction mixture was filtered, and then evaporated toluiserjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj
  • Step 1 The preparation method is the same as the compound 3a-4 except that the compound 3a-3 in the 3a-4 process is replaced with the compound 5a-4-2.
  • Step 2 The preparation method is the same as the compound 5a-6 except that the compound 5a-5 in the process of 5a-6 is replaced with the compound 13a-1.
  • Step 3 To compound 13a-2 (110 mg, 0.35 mmol) and trimethylcyclotriboroxane (0.15 ml, 1.05 mmol), cesium carbonate (228 mg, 0.7 mmol) in dioxane/water (5 ml/0.2 A catalytic amount of Pd(dppf)Cl2 was added to the mixed solution of ml), and the reaction was stirred at 90 ° C for 10 hours under a nitrogen atmosphere, and LC-MS was traced until the end of the reaction. The reaction mixture was filtered, EtOAc EtOAcjjjjjjjjjjjjjj
  • Step 4 Palladium on carbon (7 mg) was added to a solution of Compound 13a-3 (70 mg, 0.28 mmol) in methanol (5 ml). The reaction mixture was filtered, and then evaporated toluiserjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj
  • Step 1 To a solution of the compound 14a-1 (2.5 g, 7.88 mmol) in ethanol (50 ml), sodium ethoxide (1.34 g, 19.69 mmol), and the mixture was stirred at 80 ° C for 5 h. The product was poured into water, filtered, and then filtered to give white crystals, Compound Compound Compound Compound Compound Compound Compound Compound Compound Compound Compound Compound Compound Compound
  • Step 2 To a solution: 14a-2 (1.9 g, 7.11 mmol) of EtOAc (EtOAc m. The reaction solution was cooled to room temperature and then filtered, and then evaporated to ethylamine.
  • Step 3 Compound 14a-3 (3.2 g, 12.81 mmol) and 14a-4 (2.0 g, 12.81 mmol) were stirred in dichloromethane/acetic acid solution (100 ml / 2 ml) at room temperature for 30 min. Sodium hydride (8.14 g, 36.42 mmol), and the mixture was stirred at room temperature overnight. The reaction mixture was poured into water, extracted with methylene chloride. EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj ESI): 242.5 [M+H]+.
  • Step 4 Hydrochloric acid (7M, 20 ml) was added to a solution of Compound 14a-5 (2.1 g, 8.7 mmol) in tetrahydrofuran (20 ml), and the mixture was stirred at 70 ° C overnight. The reaction mixture was concentrated under reduced pressure to dryness to dryness. 1.4 g, 81%), MS m/z (ESI): 198.3 [M+H]+.
  • the preparation method is the same as the preparation method of the intermediate 14a, and 14a-3 is replaced with the corresponding amine.
  • Step 1 To a solution of the compound 14a-4 (1.36 g, 8.71 mmol) and the compound 22a-1 (900 mg, 7.26 mmol) of 1,2-dichloroethane, sodium acetate (1.48 g, 10.9 mmol). After 20 min, sodium borohydride (4.6 g, 21.8 mmol) was added, and the mixture was stirred at room temperature overnight, and LC-MS was followed until the end of the reaction. The reaction mixture was quenched with EtOAc EtOAc EtOAc (EtOAc) (1.4 g, 85%), MS m/z (ESI): 228 [M+H]+.
  • EtOAc EtOAc EtOAc
  • Step 1 Compound 14a-4 (1 g, 6.4 mmol), Compound 23a-1 (0.83 g, 6.4 mmol) and DIEA (0.82 g, 6.4 mmol) in methanol (20 ml) Sodium (2.7 g, 12.8 mmol) was stirred at room temperature overnight and was taken &lt The reaction was quenched with EtOAc EtOAc (EtOAc m. g), MS m/z (ESI): 234 [M+H]+.
  • Step 1 To a solution of the compound 24a-1 (8.6 g, 50.5 mmol) in hexanes, hexane (3.86 ml, 75.8 mmol) and p-toluenesulfonic acid (170 mg, 1.01 mmol). The reaction mixture was concentrated and purified by EtOAc EtOAcjjjj
  • Step 2 LDA (2M, 14.1 ml) was added dropwise to a solution of the compound 24a-2 (4.05 g, 18.9 mmol) in tetrahydrofuran at -78 ° C. After stirring at this temperature for 30 min, iodomethane (2.35 ml, 37.8 mmol), the reaction solution was naturally warmed to room temperature, stirred for additional 2 h, and then t The reaction mixture was quenched with EtOAc (EtOAc)EtOAc.
  • EtOAc EtOAc
  • Step 3 To a solution of compound 24a-3 (3.0 g, 13.1 mmol) in ethanol / water (20 ml / 20 ml), the mixture was stirred at 60 ° C for 10 h, The reaction is over. After the reaction mixture was cooled to room temperature, the mixture was evaporated to EtOAc (EtOAc m. %).
  • Step 4 Hydrochloric acid (4M, 30 ml) was added to a solution of compound 24a-4 (2.2 g, 11 mmol) in dioxane. The mixture was stirred at 65 ° C for 3 h. After the reaction mixture was cooled to room temperature, EtOAc was evaporated.
  • Step 5 To a solution of the compound 24a-5 (1 g, 6.41 mmol) in toluene was added DPPA (1.66 ml, 7.69 mmol), triethylamine (1.33 ml, 9.62 mmol) and benzyl alcohol (0.8 ml, 7.69 mmol). The liquid was refluxed for 10 h. The reaction mixture was concentrated to give compound 24a (m.
  • Example 1 4-(ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-methoxy-6-methyl-2-oxo-1,2-di) Preparation of Hydropyridin-3-yl)methyl)-1,5-dimethyl-1H-indazole-6-carboxamide (Compound p-1)
  • Step 1 Toluene (24 mg, 0.591 mmol) was added to a solution of compound 4a (100 mg, 0.296 mmol) in N, N-dimethylformamide (5 mL). 0.591 mmol), the mixture was stirred for additional 2 hours. LC-MS was followed until the reaction was complete. The reaction solution was poured into water, extracted with ethyl acetate, dried and evaporated to ethylamine. MS m/z (ESI): 354.2 [M+H] + .
  • Step 2 Compound 1-1 (50 mg, 0.14 mmol), Compound 2a (50 mg, 0.21 mmol), C 6 MoO 6 (50 mg, 0.21 mmol), Pd(dppf)Cl 2 (10 mg, 0.01 mmol) and triethylamine (60 mg, 0.57 mmol) of a 3 mL solution of N,N-dimethylformamide was subjected to microwave reaction at 140 ° C for 40 minutes under an argon atmosphere. LC-MS was followed until the reaction was complete. The reaction mixture was poured into water, and ethyl acetate was evaporated and evaporated.
  • Step 1 Compound 3-1 (20 g, 0.16 mol) in dichloromethane (200 mL) Stir at room temperature for 2 hours. Water was added to the reaction mixture, and the organic layer was separated. MS m/z (ESI): 164.1 [M+H] + .
  • Step 2 A solution of compound 3-2 (25.3 g, 0.15 mol) in 250 mL of acetic acid was then dropwisely weighed to a solution of Br 2 (25.2 g, 0.15 mol) in 50 mL of acetic acid at 10 ° C, and the mixture was stirred at 55 ° C for 16 hours. The reaction solution was filtered, water was added to the filtrate, and the mixture was filtered, and the mixture of the mixture was washed with ethanol, and the cake was dried under reduced pressure at 50 ° C to obtain 32.0 g of a yellow solid compound 3-3. MS m/z (ESI): 242 [M+H] + .
  • Step 3 A solution of compound 3-3 (32 g, 0.13 mol) in sulfuric acid (13.02 g, 0.13 mol) was added to the ice bath (14.54 g, 0.15 mol, 61%) in an argon atmosphere and the mixture was continued in an ice bath. Stir for 2 hours. LC-MS was followed until the reaction was complete. The reaction solution was poured into ice water, filtered, and the filter cake was washed with water and dried under reduced pressure at 50 ° C to obtain 40.0 g of solid compound 3-4. MS m/z (ESI): 287 [M+H] + .
  • Step 4 A solution of compound 3-4 (16 g, 35 mmol) in EtOAc. LC-MS was followed until the reaction was complete. Water was added to the reaction mixture, and the mixture was filtered, washed with ethanol, and dried under reduced pressure at 50 ° C to give 5.0 g of Compound 3-5. MS m/z (ESI): 245 [M+H] + .
  • Step 5 The preparation method is the same as the compound 3a-4 except that the compound 3a-3 in the 3a-4 process is replaced with the compound 3-5.
  • Step 6 The preparation method is the same as the compound 3a-2 except that the compound 3a-1 in the 3a-2 process is replaced with the compound 3-6. MS m/z (ESI): 228 [M+H] + .
  • Step 7 The preparation method is the same as the compound 4a-1 except that the compound 3a in the 4a-1 process is replaced with the compound 3-7.
  • Step 8 The preparation method is the same as the compound 4a except that the compound 4a-1 in the process of 4a is replaced with the compound 3-8.
  • Step 9 The preparation method is the same as the compound p-1 except that the compound 1-1 in the p-1 process is replaced with the compound 3-9. Purification by Prep-HPLC gave white solid compound p-3 (4 mg, 2%). MS m/z (ESI): 454.3 [M+H] + ; 1H NMR (400 MHz, DMSO-6) ⁇ 12.81 (s, 1H), 11.39 (s, 1H), 8.41 (s, 1H), 8.00 ( s,1H), 7.83(t,1H), 7.43(s,1H),6.07(s,1H), 4.20(d,2H),3.93-3.61(m,5H), 3.29-3.10(m,5H) , 2.37 (s, 3H), 2.15 (s, 3H), 1.83-1.80 (m, 1H), 1.47-1.44 (m, 1H), 1.29-1.23 (m, 2H), 0.74 (t, 3H).
  • Step 2 The preparation method is the same as the compound p-1 except that the compound 1-1 in the p-1 process is replaced with the compound 4-1. Purification by Prep-HPLC gave white solid compound p-4 (2mg, 2%). MS m/z (ESI): 468.3 [M+H] + ; 1 H NMR (400 MHz, DMSO-6) ⁇ 11.39 (s, 1H), 8.25 (s, 1H), 7.80 (t, 1H), 7.36 (s, 1H), 6.07 (s, 1H), 4.19 (d, 2H), 4.08 (s, 3H), 3.92-3.59 (m, 6H), 3.53-3.46 (m, 2H), 3.13-3.00 (m , 2H), 2.32 (s, 3H), 2.15 (s, 3H), 1.54-1.43 (m, 1H), 1.33-1.02 (m, 3H), 0.70 (t, 3H).
  • Step 1 The preparation method is the same as that of the compound 1-1 except that the compound iodomethane in the 1-1 process is replaced with sodium difluorochloroacetate to obtain a mixture of the compounds 5-1 and 5-2.
  • Step 2 The preparation method is the same as the compound p-1 except that the compound 1-1 in the p-1 process is replaced with a mixture of the compounds 5-1 and 5-2. Purification by Prep-HPLC gave white solid compound p-5 (10 mg, 6.4%).
  • 1 H NMR (400 MHz, DMSO-6) ⁇ 11.40 (s, 1H), 8.91 (s, 1H), 8.16-7.86 (m) , 2H), 7.31 (s, 1H), 6.07 (s, 1H), 4.21 (d, 2H), 3.86-3.66 (m, 5H), 3.24-3.10 (m, 5H), 2.26 (s, 3H), 2.15 (s, 3H), 1.92-1.75 (m, 1H), 1.56-.124 (m, 3H), 0.74 (t, 3H).
  • Example 50 4-(Ethyl(tetrahydro-2H-pyran-4-yl)amino)-N-((4-methoxy-6-methyl-2-oxo-1,2-di) Preparation of Hydropyridin-3-yl)methyl)-2,5-dimethyl-2H-indazole-6-carboxamide (Compound p-50)
  • the preparation method is the same as that of P-6, wherein the step 1 is carried out by referring to the method of the compound 1-1.
  • Example 8 N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-(ethyl(1-(2,2, Preparation of 2-trifluoroethyl)piperidin-4-yl)amino)-1,5-dimethyl-1H-indazole-6-carboxamide (Compound p-8)
  • Step 1 The preparation method is the same as the compound 4a-1 except that the tetrahydropyrone in the 4a-1 process is replaced by the compound 8.1. MS m/z (ESI): 390 [M+H] + .
  • Step 2 The preparation method is the same as that of the compound 4a except that the compound 4a-1 in the process of 4a is replaced with the compound 8-1.
  • Step 3 The preparation method is the same as that of the compound 1-1 except that the compound 4a in the 1-1 process is replaced with the compound 8-2.
  • Step 4 The preparation method is the same as that of the compound p-1, except that the compounds 1-1 and 2a in the p-1 process are replaced with the compounds 8-3 & 8-4 and 1a. Purification by Prep-HPLC gave white solid compound p-8 (8 mg, 8%).
  • Step 1 The preparation method is the same as the compound 4a-1 except that the tetrahydropyrone in the 4a-1 process is replaced with the compound 9.1.
  • Step 2 The preparation method is the same as that of the compound 4a, except that the compound 4a-1 in the process of 4a is replaced with the compound 9-1.
  • Step 3 The preparation method is the same as that of the compound 1-1, except that the compound 4a in the 1-1 method is replaced with the compound 9-2.
  • Step 4 The preparation method is the same as the compound p-1 except that the compounds 1-1 and 2a in the p-1 process are replaced with the compounds 9-3 and 1a. Purification by Prep-HPLC gave white solid compound p-9 (4mg, 3.6%). MS m/z (ESI): 537.1 [M+H] + ; 1 H NMR (400 MHz, DMSO-6) ⁇ 11.44 (s, 1H), 8.09 (t, 1H), 8.02 (s, 1H), 7.26 (s, 1H), 5.84 (s, 1H), 4.28 (d, 2H), 3.95 (s, 3H), 3.29-3.27 (m, 4H), 3.17 (s, 3H), 2.94-2.85 (m, 1H) ), 2.45-2.41 (m, 2H), 2.24 (s, 3H), 2.20 (s, 3H), 2.10 (s, 3H), 2.08 (s, 3H), 1.70-1.54 (m, 3H), 1.35- 1.05 (m, 5H), 0.73 (
  • Example 10 1-(Difluoromethyl)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-(B Preparation of (tetrahydro-2H-pyran-4-yl)amino)-5-methyl-1H-indazole-6-carboxamide (Compound p-10)
  • the preparation method is the same as that of the compound p-1, except that the compounds 1-1 and 2a in the process of the p-1 are replaced with the compounds 5-1 and 1a. Purification by Prep-HPLC gave white solid compound p-10 (16 mg, 16%).
  • Example 11 4-(Ethyl(1-(2,2,2-trifluoroethyl)piperidin-4-yl)amino)-N-((4-methoxy-6-methyl-2) -Oxo-1,2-dihydropyridin-3-yl)methyl)-1,5-dimethyl-1H-indazole-6-carboxamide (Compound p-11)
  • Example 12 4-(Ethyl(1-(2,2,2-trifluoroethyl)piperidin-4-yl)amino)-N-((4-methoxy-6-) Preparation of yl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-2,5-dimethyl-2H-indazole-6-carboxamide (compound p-12)
  • the preparation method is the same as that of the compound p-1, except that the compound 1-1 in the p-1 process is replaced with a mixture of the compounds 8-3 and 8-4.
  • Example 13 N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-4-(((1S,4S)-4-(((1S,4S)-4-) Preparation of dimethylamino)cyclohexyl)(ethyl)amino)-1,5-dimethyl-1H-indazole-6-carboxamide (compound p-13)
  • Step 1 To a solution of the compound 13-1 (8 g, 35.3 mmol) in anhydrous methanol (100 mL), chlorosulfoxide (7.7 mL, 106.1 mmol). TLC is tracked until the end of the reaction. After the reaction mixture was cooled, the solvent was concentrated to remove most of the solvent, filtered, and the filter cake was dried under reduced pressure to give 7.5 g of Compound 13-2. MS m/z (ESI): 242 [M+H] + .
  • Step 2 Compound 13-2 (7.6 g, 31.64 mmol) in acetic acid (150 mL) was added portionwise to iron powder (14 g, 253 mmol). After the addition, the reaction mixture was filtered, and the filtrate was poured into water and ethyl acetate Concentration and purification by combiflash gave yellow solid compound 13-3 (3.5 g, 52.6%). MS m/z (ESI): 211 [M+H] + .
  • Step 3 A solution of compound 13-3 (1 g, 4.46 mmol) in EtOAc (20 mL). LC-MS was followed until the end of the reaction. The reaction solution was poured into water, filtered, and the filter cake was dried under reduced pressure to give a white solid. MS m/z (ESI): 290.8 [M+H] + .
  • Step 4 The preparation method is the same as the compound 3a-4 except that the compound 3a-3 in the 3a-4 process is replaced with the compound 13-4.
  • Step 5 The preparation method is the same as that of the compound 1-1 except that the compound 4a in the 1-1 method is replaced with the compound 13-5. MS m/z (ESI): 315.8 [M+H] + .
  • Step 6 Compound 13-6 (900 mg, 2.87 mmol), Methylboronic acid (343 mg, 5.73 mmol), Pd (dppf) Cl 2 (210 mg, 0.286 mmol), sodium carbonate (607 mg, 5.73 mmol), 1,4- A mixture of dioxane (20 mL) and 2 mL of water was subjected to microwave reaction at 100 ° C for 8 hours under an argon atmosphere. LC-MS was followed until the reaction was complete. The reaction mixture was poured into water, and ethyl acetate was evaporated and evaporated to ethylamine. MS m/z (ESI): 249.9 [M+H] + .
  • Step 7 The preparation method was the same as that of the compound 13-3 except that the compound 13-2 in the 13-3 process was replaced with the compound 13-7, and the mixture was heated to 90 ° C and stirred for 7 hours. MS m/z (ESI): 220 [M+H] + .
  • Step 8 The preparation method is the same as the compound 4a-1 except that the tetrahydropyrone in the 4a-1 process is replaced with 4-dimethylaminocyclohexanone. MS m/z (ESI): 345.1 [M+H] + .
  • Step 9 The preparation method is the same as that of the compound 4a, except that 4a-1 in the process of 4a is replaced with the compound 13-9.
  • Step 11 To a solution of compound 13-11 (80 mg, 0.223 mmol) in EtOAc (3 mL), EtOAc (EtOAc, EtOAc, EtOAc, EtOAc Stir overnight. LC-MS was followed until the end of the reaction. The reaction mixture was extracted with EtOAc EtOAc EtOAc (EtOAc:EtOAc.
  • Example 15 4-(Ethyl(4-((2-methoxyethyl))(methyl)amino)cyclohexyl)amino)-N-((4-methoxy-6-methyl-2) -Oxo-1,2-dihydropyridin-3-yl)methyl)-1,5-dimethyl-1H-indazole-6-carboxamide (Compound p-15)
  • Step 1 The preparation method is the same as the compound 4a-1 except that the tetrahydropyrone in the 4a-1 process is replaced with the compound 15.1.
  • Step 2 The preparation method is the same as that of the compound 4a, except that the compound 4a-1 in the process of 4a is replaced with the compound 15-1.
  • Step 3 The preparation method is the same as the compound 13-11 except that the compound 13-10 in the 13-11 process is replaced with the compound 15-2.
  • Step 4 The preparation method is the same as the compound p-13 except that the compound 13-11 in the p-13 process is replaced with the compound 15-3. Purification by Prep-HPLC gave white solid compound p-15 (25 mg, 10%). MS m/z (ESI): 553.3 [M+H] + ; 1 H NMR (400 MHz, DMSO-6) ⁇ 11.39 (s, 1H), 8.02 (s, 1H), 7.78 (t, 1H), 7.25 (s, 1H), 6.07 (s, 1H), 4.23 (d, 2H), 3.97 (s, 3H), 3.79 (s, 3H), 3.51-3.41 (m, 2H), 3.29-3.27 (m, 2H) ), 3.20-3.15 (m, 4H), 2.94-2.84 (m, 1H), 2.46-2.43 (m, 2H), 2.24 (s, 3H), 2.15 (s, 3H), 2.10 (s, 3H), 1.72-1.57 (m, 3H), 1.33-1.07 (m
  • Step 1 The preparation method is the same as the compound 13-7 except that the methylboronic acid in the 13-7 process is replaced with the compound 16.1. MS m/z (ESI): 2621. [M+H] + .
  • Step 2 Compound 16-1 (50 mg, 0.2 mmol) in MeOH (5 mL) LC-MS was followed until the reaction was complete. The reaction mixture was filtered and concentrated to give compound 16-2. MS m / z (ESI): 234.2 [M + H] +.
  • Step 3 The preparation method is the same as the compound 4a-1 except that the compound 3a in the 4a-1 process is replaced with the compound 16-2.
  • Step 4 The preparation method is the same as that of the compound 4a, except that the compound 4a-1 in the process of 4a is replaced with the compound 16-3.
  • Step 5 The preparation method is the same as the compound 13-11 except that the compound 13-10 in the 13-11 process is replaced with the compound 16-4. MS m/z (ESI): 332.2 [M+H] + .
  • Step 6 The preparation method is the same as the compound p-13 except that the compound 13-11 in the p-13 process is replaced with the compound 16-5. Purification by Prep-HPLC gave white solid compound p-16 (46 mg, 16%). MS m/z (ESI): 482.2 [M+H] + ; 1 H NMR (400 MHz, DMSO) ⁇ 11.41 (s, 1H), 8.07 (s, 1H), 7.92 (t, 1H), 7.31 (s) ,1H),6.07(s,1H), 4.26(d,2H),3.95(s,3H),3.83-3.71(m,2H),3.78(s,3H),3.24-3.18(m,5H), 2.96-2.92 (m, 2H), 2.17 (s, 3H), 1.85 (s, 1H), 1.55 (s, 1H), 1.38-1.29 (m, 2H), 0.99 (t, 3H), 0.77 (t, 3H).
  • Example 18 N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-ethyl-4-(ethyl ((1S) ,4S)-4-(3-Methylazetidin-1-yl)cyclohexyl)amino)-1-methyl-1H-indazole-6-carboxamide (Compound p-18)
  • Step 1 The preparation method is the same as that of the compound 4a-1, except that the compound 3a and the tetrahydropyrone in the 4a-1 process are replaced with the compounds 16-2 and 18.1. MS m/z (ESI): 399 [M+H] + .
  • Step 2 The preparation method is the same as that of the compound 4a, except that the compound 4a-1 in the process of 4a is replaced with the compound 18-1.
  • Step 3 The preparation method is the same as the compound 13-11 except that the compound 13-10 in the 13-11 process is replaced with the compound 18-2.
  • Step 4 The preparation method is the same as the compound p-13 except that the compounds 13-11 and 2a in the p-13 process are replaced with the compounds 18-3 and 1a. Purification by Prep-HPLC to give white solid compound p-18; 1 H NMR ( 400MHz, DMSO) ⁇ 11.46 (s, 1H), 8.13 (t, 1H), 8.05 (s, 1H), 7.30 (s, 1H) , 5.87 (s, 1H), 4.30 (d, 2H), 3.97 (s, 3H), 3.86-3.82 (m, 1H), 3.38 (t, 2H), 3.11 (s, 3H), 2.93 (t, 4H) ), 2.67 (t, 2H), 2.36-2.29 (m, 1H), 2.23 (s, 3H), 2.11 (s, 3H), 1.89-1.55 (m, 6H), 0.96 (t, 3H), 0.77 ( t, 6H) with p-19;.
  • Example 20 N-((4,6-Dimethyl-2-oxo-1,2-dihydropyridin-3-yl)methyl)-5-ethyl-4-(ethyl ((1S) Of 4S)-4-((2-methoxyethyl)(methyl)amino)cyclohexyl)amino)-1-methyl-1H-indazole-6-carboxamide (Compound p-20)
  • Step 1 The preparation method is the same as that of the compound 4a-1, except that the compound 3a and the tetrahydropyrone in the 4a-1 process are replaced with the compounds 16-2 and 20.1.
  • Step 2 The preparation method is the same as the compound 4a except that the compound 4a-1 in the process of 4a is replaced with the compound 20-1.
  • Step 3 The preparation method is the same as the compound 13-11 except that the compound 13-10 in the 13-11 process is replaced with the compound 20-2.
  • Step 4 The preparation method is the same as the compound p-13 except that the compounds 13-11 and 2a in the p-13 process are replaced with the compounds 20-3 and 1a. Purified by Prep-HPLC to give white solid compound by p-20; 1 H NMR (400MHz, DMSO -6) ⁇ 11.43 (s, 1H), 8.10 (t, 1H), 8.05 (s, 1H), 7.30 (s, 1H), 5.84 (s, 1H), 4.28 (d, 2H), 3.96 (s, 3H), 3.54-3.47 (m, 2H), 3.25 (s, 3H), 3.21-2.77 (m, 8H), 2.67 -2.50 (m, 3H), 2.20 (s, 3H), 2.08 (s, 3H), 1.98-1.71 (m, 3H), 1.62-1.52 (m, 1H), 1.44-1.21 (m, 4H), 0.95 (t, 3H), 0.76 (t, 3H). MS m/z (ESI): 551.3 [M+H]
  • Example 22 4-((4-(Dimethylamino)cyclohexyl)(ethyl)amino)-5-ethyl-N-((4-methoxy-6-methyl-2-oxo) Of -1,2-dihydropyridin-3-yl)methyl)1-methyl-1H-indazole-6-carboxamide (P-22)
  • Step 1 Compound 5a (100 mg, 0.408 mmol), 4-dimethylcyclohexanone (121 mg, 0.857 mmol) in dioxane / trifluoroacetic acid (10 ml / 2 ml) mixture was stirred at room temperature for 2 h, and acetic acid was added. Sodium borohydride (272 mg, 1.29 mmol) was added and the mixture was stirred at room temperature for 1 h and then filtered and evaporated and evaporated. The reaction was quenched with EtOAc EtOAc (EtOAc m. (ESI): 359.3 [M+H] + .
  • Step 2 Compound 22-1 (150 mg, 0.418 mmol), acetaldehyde (92 mg, 2.07 mmol) in dioxane/acetic acid (20 ml / 2 ml) was stirred at room temperature for 1 h and sodium borohydride (443 mg) , 2.09 mmol), the mixture was stirred at room temperature for 1 h, and LC-MS was followed until the end of the reaction. The reaction was quenched with EtOAc EtOAc (EtOAc m. + H] +.
  • Step 3 A solution of the compound 22-2 (150 mg, 0.388 mmol) in methanol was then evaporated. The reaction mixture was concentrated under reduced pressure of EtOAc (EtOAc). Obtained as a black oil 22-3 (150 mg), MS m/z (ESI): 373.4 [M+H] + .
  • Step 4 To a solution of compound 22-3 (140 mg, 0.576 mmol) in EtOAc (mjjjjjjjjjjjjjj LC-MS was traced to the end of the reaction. The reaction mixture was poured into water and extracted with EtOAc. EtOAc (EtOAc m. m / z (ESI): 523.3 [m + H] +.
  • the target compound of the examples is represented by the formula (II), and the substituents R 1 , R 11 , R 12 and Y are shown in the following table.
  • Step 1 The preparation method was the same as that of the compound 22-1 except that the compound 5a and 4-dimethylcyclohexanone were replaced with the compounds 8a and 14a-4 at a reaction temperature of 0 °C.
  • Step 2 The preparation method was the same as the compound 22-2 except that the compound 22-1 was changed to 44-1, and the reaction temperature was 0 °C. MS m/z (ESI): 388.3 [M+H] + .
  • Step 3 A solution of compound 44-2 (750 mg, 1.94 mmol) elute After the reaction mixture was cooled to room temperature, the mixture was evaporated, evaporated, evaporated, evaporated , 97%), MS m / z (ESI): 344.3 [m + H] +.
  • Step 4 Compound 44-3 (500 mg, 1.46 mmol) and compound 44.1 (288 mg, 2.91 mmol) in methylene chloride/acetic acid (10ml / 0.2ml) were stirred at 0 ° C for 1 h. Sodium (925 mg, 4.37 mmol) was stirred at 0<0>C for 2 h. The reaction was quenched with EtOAc EtOAc (EtOAc m. /z(ESI):427.3[M+H] +.
  • Step 5 The preparation method is the same as the compound 44-4 except that the compounds 44-3 and 44.1 are replaced with the compound 44-4 and formaldehyde, and the solvent is changed to a methanol/acetic acid solution.
  • Step 6 The preparation method is the same as the compound 22-3 except that the compound 22-2 is replaced with 44-5. MS m/z (ESI): 427.3 [M+H] + .
  • Step 7 The preparation was carried out in the same manner as the compound P-22 except that the compounds 22-3 and 2a were replaced with the compounds 44-6 and 1a. Purified by Pre-HPLC to give white solid compound P-44 (54mg, 37% ), MS m / z (ESI): 561.3 [M + H] +.
  • Example 45 4-((4-((2,2-Difluoroethyl)(methyl)amino)cyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2) Of oxo-1,2-dihydropyridin-3-yl)methyl)-1,5-dimethyl-1H-indazole-6-carboxamide (P-45)
  • Example 46 4-((4-Amino-4-methylcyclohexyl)(ethyl)amino)-N-((4,6-dimethyl-2-oxo-1,2-dihydropyridine) Preparation of -3-yl)methyl)-1H-indazole-6-carboxamide (P-46)
  • Step 1 to Step 4 The preparation method is the same as the compound P-22, except that the compound 5a and 2-dimethylcyclohexanone in the step 1 are replaced with the compounds 8a and 24a.
  • Step 5 To a solution of compound 46-4 (180 mg, 0.29 mmol) in EtOAc/EtOAc (EtOAc) End. The reaction was filtered, the filtrate was concentrated, purified by Pre-HPLC to give white solid compound P-46 (19.11mg, 14% ), MS m / z (ESI): 479.3 [M + H] +.
  • Step 1 to Step 4 The preparation method is the same as the compound P-22 except that the compound 2-dimethylcyclohexanone in the step 1 is replaced with the compound 22a.
  • Step 5 To a solution of the compound 47-4 (30 mg, 0.055 mmol) in methylene chloride (methanol) (3M, The MS tracks until the end of the reaction. Methanol (10 ml) was added to the reaction mixture, and the mixture was evaporated. , 24%), MS m/z (ESI): 535.3 [M+H] + .
  • methanol methylene chloride
  • Step 1 The preparation was carried out in the same manner as in the compound 22-1 except that the compound 5a and 4-dimethylcyclohexanone were replaced with the compounds 8a and 48.1.
  • Step 2 The preparation method is the same as the compound 22-2 except that the compound 22-1 is replaced by 48-1.
  • Step 3 To a solution of compound 48-2 (358 mg, 0.77 mmol) in methanol, EtOAc (EtOAc) (EtOAc) The reaction was filtered, the filtrate was concentrated to afford compound 48-3 (240mg), MS m / z (ESI): 331 [M + H] +.
  • Step 4 Compound 48-3 (240 mg, 0.72 mmol),jjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjjj
  • Step 5 The preparation method is the same as the compound 22-3 except that the compound 22-2 is replaced with the compound 48-4. MS m/z (ESI): 373 [M+H] + .
  • Step 6 The preparation method is the same as the compound 22-4 except that the compounds 22-3 and 2a are replaced with the compounds 48-4 and 1a. MS m/z (ESI): 507 [M+H] + .
  • Step 1 The preparation method is the same as that of the compound 22-1, except that the compound 5a and 4-dimethylaminocyclohexanone in the preparation are replaced with the compound 13a and the tetrahydropyrone.
  • Step 2 The preparation method is the same as that of the compound 22-2, except that the compound 22-1 in the process is replaced with 49-1. MS m/z (ESI): 332.8 [M+H] + .
  • Step 3 The preparation method is the same as the compound 22-3 except that the compound 22-2 in the process is replaced with 49-2. MS m/z (ESI): 318.3 [M+H] + .
  • Step 4 The preparation method is the same as the compound P-22 except that the compounds 22-3 and 2a in the preparation are replaced with the compounds 49-3 and 1a.
  • Recombinant PRC2 (EZH2-Y641F) was purchased from Active Motif, S-methylthioadenosine (SAM) and L-polylysine (PLL) were purchased from Sigma-Aldrich, H3(1-50)K27me1 polypeptide. Purchased from Cisbio.
  • the detection system uses Perkinelmer's LANCEUltra system. In the enzyme activity experiment, the test compound was diluted at a gradient of 1:3, and then added to the reaction plate and 100 ng of the recombinase was added.
  • the cell lines used, Pfeiffer (CRL-2632), suDHL-6 (CRL-2959), and suDHL-10 (CRL-2963) were purchased from the American Type Culture Collection (ATCC). All cell lines were cultured in RPMI-1640 medium (Gbico) containing 10% fetal bovine serum (Gibco). The cultured cells were collected by centrifugation and the cell density was measured on a CounterStar counter. The appropriate number of cells were then seeded in 96-well plates and incubated overnight. The test compound was diluted at 8 gradient points in a ratio of 1:3 and added to the corresponding wells.
  • the type of benzo five-membered heterocyclic ring has a great influence on the inhibitory activity of the enzyme, and when the compound B-1 is substituted into the formula B-2, the formula B-3 or the formula B-4 (for example, P-3, P) -4 or P-50 and P-7), the inhibitory activity against the enzyme was significantly reduced.
  • the compound structure is of the formula B-1, whether or not the 3-position has a substituent, and the change of the 5-position substituent of the benzene ring has a large influence on the inhibitory activity of the enzyme, when the hydrogen at the 3-position is methyl or After fluorine substitution (such as P-42 and P-13/14, P-41 and P-7), the inhibitory activity on the enzyme is greatly reduced.
  • the 5-position methyl group of the benzene ring is replaced with other substituents such as cyano group, cyclopropyl group At the time of the base, the inhibitory activity against the enzyme was significantly lowered.

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Abstract

提供一种4,5,6-三取代吲唑类衍生物、其制备方法与医药上的用途，具体地，提供一种式（I）化合物或其药学上可接受的盐、立体异构体、溶剂化物或前药，及其制备方法和应用。

Description

4,5,6-三取代吲唑类衍生物、其制法与医药上的用途 技术领域
本发明属于医药技术领域。具体地，本发明特别涉及一种4,5,6-三取代吲唑类衍生物及其制备方法和作为EZH2抑制剂的应用，以及由其制备的药物组合物。
背景技术
组蛋白-赖氨酸-N-甲基转移酶EZH2参与DNA的甲基化和最终的转录抑制；通过辅因子S-腺苷-L-甲硫氨酸催化赖氨酸27位的甲基转移至组氨酸H3。这种甲基化促进异染色质的形成，从而引发基因沉默。
EZH2是PRC2功能酶的部分，是通过表观遗传学维持控制调节发育和分化的基因，从而保证胚胎健康发育。EZH2的突变或过表达与许多癌症的形成相关。EZH2控制基因控制肿瘤发展，抑制EZH2的活性会减慢肿瘤的生长速度。作为靶向抑制剂，EZH2能调控多种癌症包括，乳腺癌，前列腺癌，黑色素瘤和膀胱癌。
PCT申请WO2011140324A1和WO2012075080A1披露了吲哚类化合物作为EZH2抑制剂用于治疗癌症。PCT申请WO2012118812A2公开了双环杂环化合物作为EZH2抑制剂用于癌症的治疗。
因此，抑制EZH2活性将有效降低细胞增殖和侵袭，从而为EZH2介导的疾病或病症提供有益的治疗。本发明化合物作为EZH2抑制剂为疾病或由EZH2介导的肿瘤治疗提供解决方案。
发明内容
本发明的目的是提供一种结构新颖的可作为EZH2抑制剂的化合物。
本发明第一方面提供了一种式(I)所示的化合物，或其药学上可接受的盐、溶剂化物、立体异构体或前药：
Figure PCTCN2017110465-appb-000001
式中，
R0为氢；
R1为CN、卤素(优选氟、氯、溴)、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C1-8烷氧基(优选为C1-6烷氧基，更优选为C1-3烷氧基)、C3-8环烷基(优选为C3-6环烷基)或C3-8环烷氧基(优选为C3-6环烷氧基)；
R2为氢、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)或C3-8环烷基(优选为C3-6环烷基)；
R3为氢、卤素(优选氟、氯、溴)、羟基、CN、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C3-8环烷基(优选为C3-6环烷基)、C1-8烷氧基(优选为C1-6烷氧基，更优选为C1-3烷氧基)、卤代C1-8烷氧基(优选为卤代C1-6烷氧基，更优选为卤代C1-3烷氧基)、C3-8环烷氧基(优选为C3-6环烷氧基)、C6-10芳基(优选苯基)、-C(O)C1-8烷基(优选为-C(O)C1-6烷基，更优选为-C(O)C1-3烷基)、-C(O)OC1-8烷基(优选为-C(O)OC1-6烷基，更优选为-C(O)OC1-3烷基)、-CONRaRb或NRaRb；
X为NR4、CR5R6、O、S或S(O)2；
R4为氢、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C(O)C1-8烷基(优选为C(O)C1-6烷基，更优选为C(O)C1-3烷基)、C(O)OC1-8烷基(优选为C(O)OC1-6烷基，更优选为C(O)OC1-3烷基)、CONRa1Rb1、C3-8环烷基(优选为C3-6环烷基)、-SO2C1-10烷基(优选为-SO2C1-6烷基，更优选为-SO2C1-3烷基)、-C(O)CH2CN、-C(O)CH2OH、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环；
R5、R6各自独立地为氢、卤素、羟基、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C3-8环烷基(优选为C3-6环烷基)、NRa2Rb2、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；
Ra、Rb、Ra1、Rb1、Ra2、Rb2各自独立地为氢、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、C(O)C1-8烷基(优选为C(O)C1-6烷基，更优选为C(O)C1-3烷基)；
A环为式(A-1)、式(A-2)、式(A-3)或式(A-4)所示结构：
Figure PCTCN2017110465-appb-000002
其中，R11、R21、R31、R41各自独立地为氢、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C3-8环烷基(优选为C3-6环烷基)、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；
R12、R22、R32、R42各自独立地为氢、卤素(优选氟、氯、溴)、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)或C3-8环烷基(优选为C3-6环烷基)；
所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1、2或3个选自下组的取代基所取代：CN、乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OC1-6烷基、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、C1-8烷氧基(优选为C1-6烷氧基，更优选为C1-3烷氧基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C3-8环烷基(优选为C3-6环烷基)、卤代C1-8烷氧基(优选为卤代C1-6烷氧基，更优选为卤代C1-3烷氧基)、NRa3Rb3、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环；其中Ra3、Rb3 各自独立地为氢、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)或C1-8烷氧基取代的C1-8烷基(优选为C1-6烷氧基取代的C1-6烷基，更优选为C1-3烷氧基取代的C1-3烷基)。
在另一优选例中，R1为C1-3烷基(优选为甲基或乙基)。
在另一优选例中，R2为氢、C1-3烷基或卤代C1-3烷基。
在另一优选例中，R2为C1-3烷基(优选甲基或乙基)。
在另一优选例中，R3为氢、卤素、C1-3烷基或C1-3烷氧基。
在另一优选例中，R3为氢、氟、氯、甲基、乙基、正丙基、异丙基、甲氧基、乙氧基、丙氧基、异丙氧基。
在另一优选例中，R0为氢且R1为C1-3烷基(优选为甲基或乙基)。
在另一优选例中，R11、R21、R31、R41各自独立地为氢或-(CH2)n-L1；其中L1为CN、NRa3Rb3、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、C1-8烷氧基(优选为C1-6烷氧基，更优选为C1-3烷氧基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C3-8环烷基(优选为C3-6环烷基)、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；n为0、1或2；
所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OC1-6烷基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、NRa3Rb3、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃；Ra3、Rb3如前所定义。
在另一优选例中，n为0或1。
在另一优选例中，所述4至6元饱和单杂环为氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、哌嗪、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃。
在另一优选例中，所述5至6元单环杂芳基环为噻吩、N-烷基吡咯、呋喃、噻唑、咪唑、噁唑、吡咯、吡唑、三唑、四唑、异噁唑、噁二唑、噻二唑、吡啶、哒嗪、嘧啶或吡嗪。
在另一优选例中，取代基中的氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃为未取代的或被1、2或3个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、卤代C1-3烷氧基、-C(O)OC1-6烷基、NRa3Rb3；其中Ra3、Rb3各自独立地为氢或C1-3烷基。
在另一优选例中，R11、R21、R31、R41各自独立地为氢、甲基、乙基、正丙基、异丙基、环丙基、一氟甲基、二氟甲基、三氟甲基、一氟乙基、二氟乙基、三氟乙基。
在另一优选例中，R12、R22、R32、R42各自独立地为氢、甲基、乙基、正丙基、异丙基。
在另一优选例中，R12、R22、R32、R42各自独立地为氢。
在另一优选例中，R4为氢或-(CH2)m-L2；其中L2为CN、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、NRa3Rb3、C1-8烷氧基(优选为C1-6烷氧基，更优选为C1-3烷氧基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C(O)C1-8烷基(优选为C(O)C1-6烷基，更优选为C(O)C1-3烷基)、C(O)OC1-8烷基(优选为C(O)OC1-6烷基，更优选为C(O)OC1-3烷基)、CONRa1Rb1、C3-8环烷基(优选为C3-6环烷基)、-SO2C1-8烷基(优选为-SO2C1-6烷基，更优选为-SO2C1-3烷基)、-C(O)CH2CN、-C(O)CH2OH、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳 基环、螺环、螺杂环、桥环或桥杂环；m为0、1或2；
所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OC1-6烷基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、NRa3Rb3、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃；Ra3、Rb3如前所定义。
在另一优选例中，m为0或1。
在另一优选例中，R4为-(CH2)m-L2；其中L2为卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)或4至6元饱和单杂环；m为0或1。
在另一优选例中，R4为-CH2-L2；其中L2为卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)。
在另一优选例中，R4为L2；其中L2为4至6元饱和单杂环。
在另一优选例中，所述4至6元饱和单杂环为氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、哌嗪、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃。
在另一优选例中，所述5至6元单环杂芳基环为噻吩、N-烷基吡咯、呋喃、噻唑、咪唑、噁唑、吡咯、吡唑、三唑、四唑、异噁唑、噁二唑、噻二唑、吡啶、哒嗪、嘧啶或吡嗪。
在另一优选例中，取代基中的氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃为未取代的或被1、2或3个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、卤代C1-3烷氧基、-C(O)OC1-6烷基、NRa3Rb3；其中Ra3、Rb3各自独立地为氢或C1-3烷基。
在另一优选例中，R4为氢、卤代C1-3烷基。
在另一优选例中，R5为氢且R6为-(CH2)p-L3；L3为CN、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、C1-8烷氧基(优选为C1-6烷氧基，更优选为C1-3烷氧基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C3-8环烷基(优选为C3-6环烷基)、NRa3Rb3、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；p为0、1或2；
所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OC1-6烷基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、NRa3Rb3、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃；Ra3、Rb3如前所定义。
在另一优选例中，p为0、1或2；L3为NRa3Rb3或4至6元饱和单杂环；Ra3、Rb3各自独立地为氢、C1-3烷基或C1-3烷氧基取代的C1-3烷基；所述烷基、烷氧基或4至6元饱和单杂环为未取代的或被1个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OCH3、-C(O)OCH2CH3、-C(O)OC(CH3)3、-C(O)OCH(CH3)2、甲基、乙基、正丙基、异丙基、甲氧基、乙氧基、丙氧基、异丙氧基、一氟甲基、二氟甲基、三氟甲基、一氟乙基、二氟乙基、三氟乙基、环丙基、环丁基、环戊基、环己基、N(CH3)2、N(CH2CH3)2、N(CH2CH3)(CH3)、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、哌嗪、二氧戊环、二氧六环、吗啉、 硫代吗啉、硫代吗啉-1,1-二氧化物、四氢吡喃。
在另一优选例中，所述4至6元饱和单杂环为氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、哌嗪、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃。
在另一优选例中，所述5至6元单环杂芳基环为噻吩、N-烷基吡咯、呋喃、噻唑、咪唑、噁唑、吡咯、吡唑、三唑、四唑、异噁唑、噁二唑、噻二唑、吡啶、哒嗪、嘧啶或吡嗪。
在另一优选例中，取代基中的氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃为未取代的或被1、2或3个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、卤代C1-3烷氧基、-C(O)OC1-6烷基、NRa3Rb3；其中Ra3、Rb3各自独立地为氢或C1-3烷基。
在另一优选例中，
Figure PCTCN2017110465-appb-000003
选自下组：
Figure PCTCN2017110465-appb-000004
在另一优选例中，
Figure PCTCN2017110465-appb-000005
为式(B-1)、式(B-2)、式(B-3)或式(B-4)所示结构：
Figure PCTCN2017110465-appb-000006
其中R11、R12、R21、R22、R31、R32、R41、R42如前所定义。
在另一优选例中，
Figure PCTCN2017110465-appb-000007
为式(B-1)所示结构。
在另一优选例中，所述化合物选自下表A：
表A
Figure PCTCN2017110465-appb-000008
Figure PCTCN2017110465-appb-000009
在另一优选例中，所述化合物选自下表B：
表B
Figure PCTCN2017110465-appb-000010
Figure PCTCN2017110465-appb-000011
本发明第二方面提供了一种药物组合物，所述药物组合物包括本发明第一方面所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药；以及药学可接受的载体。
本发明第三方面提供了如本发明第一方面所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药、或如本发明第二方面所述药物组合物在制备EZH2抑制剂的应用。
本发明第三方面提供了如本发明第一方面所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药、或如本发明第二方面所述药物组合物在制备由EZH2介导的疾病或病症的应用。
本发明第四方面提供了一种治疗由EZH2介导的疾病或病症的方法，包括给予所需患者治疗有效量的本发明第一方面所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，或如本发明第二方面所述药物组合物。
本发明第五方面提供了一种治疗由EZH2介导的疾病或病症的方法，包括给予所需患者治疗有效量的本发明第一方面所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，以及另一种治疗活性试剂。
在另一优选例中，由EZH2介导的疾病或病症选自：癌症、肺动脉高压、骨髓纤维化、人类免疫缺陷病毒(HIV)疾病、移植物抗宿主病(GVHD)、韦弗综合征、寻常性银屑病或肝纤维化。
在另一优选例中，由EZH2介导的疾病或病症为癌症。
在另一优选例中，由EZH2介导的癌症包括但不是限于，甲状腺癌、心脏肉瘤、肺癌、胃肠道癌、泌尿生殖道肿瘤、肝癌、套细胞淋巴瘤、骨肉瘤，神经系统肉瘤、妇科癌、血液系统肿瘤、肾上腺神经母细胞瘤、皮肤癌、星形细胞肿瘤、乳腺癌、大肠癌、子宫内膜癌，头颈部癌、口腔癌。
应理解，在本发明范围内中，本发明的上述各技术特征和在下文(如实施例)中具体描述的各技术特征之间都可以互相组合，从而构成新的或优选的技术方案。限于篇幅，在此不再一一累述。
具体实施方式
本发明人经过广泛而深入的研究，意外地发现了这类4,5,6-三取代吲唑类衍生物，特别是吲唑7位无取代的4,5,6-三取代吲唑类衍生物对EZH2Y641F等酶以及SU-DHL-6和SU-DHL-10等细胞具有较高的抑制活性。因此该系列化合物有望开发成为用于治疗肿瘤的药物。在此基础上，发明人完成了本发明。
术语定义
如本文所用，“烷基”指直链和支链的饱和的脂族烃基，C1-8烷基为包含1至8个碳原子的烷基，优选为C1-6烷基，更优选为C1-3烷基，定义类似；烷基的非限制性的例子包括：甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基、正庚基、2-甲基己基、3-甲基己基、4-甲基己基、5-甲基己基、2,3-二甲基戊基、2,4-二甲基戊基、2,2-二甲基戊基、3,3-二甲基戊基、2-乙基戊基、3-乙基戊基、正辛基、2,3-二甲基己基、2,4-二甲基己基、2,5-二甲基己基、2,2-二甲基己基、3,3-二甲基己基、4,4-二甲基己基、2-乙基己基、3-乙基己基、4-乙基己基、2-甲基-2-乙基戊基、2-甲基-3-乙基戊基、正壬基、2-甲基-2-乙基己基、2-甲基-3-乙基己基、2,2-二乙基戊基、正癸基、3,3-二乙基己基、2,2-二乙基己基，及其各种支链异构体等。
如本文所用，“环烷基”指饱和或部分不饱和单环环状烃基，“C3-8环烷基”是指包含3至8个碳原子的环烃基，优选为C3-6环烷基，定义类似；。环烷基的非限制性实施例包括环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基、环庚三烯基、环辛基等，优选环丙基、环戊基、环己烯基。
如本文所用，“螺环”是指单环之间共用一个碳原子(称螺原子)的多环基团，这些可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。根据环的数目将螺环分为双螺环或多螺环，优选为双螺环。更优选为优选为4元/5元、5元/5元或5元/6元双螺环。例如：
Figure PCTCN2017110465-appb-000012
如本文所用，“螺杂环”指单环之间共用一个原子(称螺原子)的多环烃，其中一个或两个环原子选自氮、氧或S(O)n(其中n是整数0至2)的杂原子，其余环原子为碳。这些可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。根据环的数目将螺杂环分为双螺杂环或多螺杂环，优选双螺杂环。更优选为4元/5元、5元/5元或5元/6元双螺杂环。例如：
Figure PCTCN2017110465-appb-000013
如本文所用，“桥环”是指共用两个或两个以上碳原子的多环基团，共用的碳原子称为桥头碳，两个桥头碳之间可以是碳链，也可以是一个键，称为桥。这些可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。优选为双环或三环桥环。例如：
Figure PCTCN2017110465-appb-000014
如本文所用，“桥杂环”指共用两个或两个以上原子的多环基团，其中一个或多个环原子选自氮、氧或S(O)n(其中n是整数0至2)的杂原子，其余环原子为碳。这些可以含有一个或多个双键，但没有一个环具有完全共轭的π电子系统。优选为双环或三环桥杂环。例如：
Figure PCTCN2017110465-appb-000015
如本文所用，“8至10元双环”是指含8至10个环原子的含两个环的桥环，双环可为饱和全碳双环或部分不饱和的全碳双环，双环的实例包括(但不限于)：
Figure PCTCN2017110465-appb-000016
如本文所用，“8至10元双杂环”是指含8至10个环原子的含两个环的桥杂环，其中1、2、3、4或5个环碳原子被选自氮、氧或硫的杂原子所取代。双杂环的实例包括(但不限于)四氢喹啉环、四氢异喹啉环、十氢喹啉环等。
如本文所用，“C1-8烷氧基”指-O-(C1-8烷基)，其中烷基的定义如上所述。优选C1-6烷氧基，更优选C1-3烷氧基。非限制性实施例包含甲氧基、乙氧基、丙氧基、异丙氧基、丁氧基、叔丁氧基、异丁氧基、戊氧基等。
如本文所用，“C3-8环烷氧基”指-O-(C3-8环烷基)，其中环烷基的定义如上所述。优选C3-6环烷氧基。非限制性实施例包含环丙氧基、环丁氧基、环戊氧基、环己氧基等。
如本文所用，“C6-10芳基”指具有共轭的π电子体系的全碳单环或稠合多环(也就是共享毗邻碳原子对的环)基团，指含有6至10个碳原子的芳基；优选苯基和萘基，更优选苯基。
如本文所用，“一个键”指由其连接的两个基团通过一个共价键连接。
如本文所用，“卤素”指氟、氯、溴或碘。
如本文所用，“卤代”指基团中一个或多个(如1、2、3、4或5个)氢被卤素所取代。
例如，“卤代C1-8烷基”指烷基被一个或多个(如1、2、3、4或5个)卤素取代，其中烷基的定义如上所述。选为卤代C1-6烷基，更优选为卤代C1-3烷基。卤代C1-8烷基的例子包括(但不限于)一氯甲基、二氯甲基、三氯甲基、一氯乙基、1,2-二氯乙基、三氯乙基、一溴乙基、一氟甲基、二氟甲基、三氟甲基、一氟乙基、二氟乙基、三氟乙基等。
又例如，“卤代C1-8烷氧基”指烷氧基被一个或多个(如1、2、3、4或5个)卤素取代，其中烷氧基的定义如上所述。优选为卤代C1-6烷氧基，更优选为卤代C1-3烷氧基。包括(但不限于)三氟甲氧基、三氟乙氧基、一氟甲氧基、一氟乙氧基、二氟甲氧基、二氟乙氧基等。
又例如，“卤代C3-8环烷基”指环烷基被一个或多个(如1、2、3、4或5个)卤素取代，其中环烷基的定义如上所述。优选为卤代C3-6环烷基。包括(但不限于)三氟环丙基、一氟环丙基、一氟环己基、二氟环丙基、二氟环己基等。
如本文所用，“氘代C1-8烷基”指烷基被一个或多个(如1、2、3、4或5个)氘原子取代，其中烷基的定义如上所述。优选为氘代C1-6烷基，更优选为氘代C1-3烷基。氘代C1-20烷基的例子包括(但不限于)单氘代甲基、单氘代乙基、二氘代甲基、二氘代乙基、三氘代甲基、三氘代乙基等。
如本文所用，“氨基”指NH2，“氰基”指CN，“硝基”指NO2，“苄基”指-CH2-苯基，“氧代基”指＝O，“羧基”指-C(O)OH，“乙酰基”指-C(O)CH3，“羟甲基”指-CH2OH，“羟乙基”指-CH2CH2OH，“羟基”指-OH，“硫醇”指SH，“亚环丙基”结构为：
Figure PCTCN2017110465-appb-000017
如本文所用，“杂芳基环”与“杂芳基”可互换使用，是指具有5到10个环原子，优选5或6元单环杂芳基或8至10元双环杂芳基；环阵列中共享6、10或14个π电子；且除碳原子外还具有1到5个杂原子的基团。“杂原子”是指氮、氧或硫。
如本文所用，“3至6元饱和或部分不饱和单环”是指含3至6个环原子的饱和或部分不饱和的全碳单环。单环的实例包括(但不限于)：环丙基环、环丁基环、环戊基环、环戊烯基环、环己基环、环己烯基环、环己二烯基环、环庚基环、环庚三烯基环、环辛基环等。
如本文所用，“3至6元饱和单杂环”是指3至6元单环中的1、2或3个碳原子被选自氮、氧或S(O)t(其中t是整数0至2)的杂原子所取代，但不包括-O-O-、-O-S-或-S-S-的环部分，其余环原子为碳；优选4至6元，更优选5至6元。饱和单杂环的实例包括(但不限于)环氧丙烷、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、吡咯啉、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物、四氢吡喃等。
如本文所用，“5至6元单环杂芳基环”是指含5至6个环原子的单杂芳基环，例如包括(但不限于)：噻吩环、N-烷基吡咯环、呋喃环、噻唑环、咪唑环、噁唑环、吡咯环、吡唑环、三唑环、四唑环、异噁唑环、噁二唑环、噻二唑环、吡啶环、哒嗪环、嘧啶环、吡嗪环等。
如本文所用，“8至10元双环杂芳基环”是指含8至10个环原子的双杂芳基环，例如包括(但不限于)：苯并呋喃、苯并噻吩、吲哚、异吲哚、喹啉、异喹啉、吲唑、苯并噻唑、苯并咪唑、喹唑啉、喹喔啉、噌啉、酞嗪。
如本文所用，“取代的”指基团中的一个或多个氢原子，优选为1～5个氢原子彼此独立地被相应数目的取代基取代，更优选为1～3个氢原子彼此独立地被相应数目的取代基取代。不言而喻，取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。
如本文所用，本文任一基团可以是取代的或未取代的。上述基团为取代时，取代基优选为1至5个以下基团，独立地选自CN、卤素、C1-8烷基(优选为C1-6烷基，更优选为C1-3烷基)、C1-8烷氧基(优选为C1-6烷氧基，更优选为C1-3烷氧基)、卤代C1-8烷基(优选为卤代C1-6烷基，更优选为卤代C1-3烷基)、C3-8环烷基(优选为C3-6环烷基)、卤代C1-8烷氧基(优选为卤代C1-6烷氧基，更优选为卤代C1-3烷氧基)、C1-8烷基取代的胺基、胺基、卤代C1-8烷基取代的胺基、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环。
本文以上所述的各类取代基团其自身也是可以被本文所描述的基团取代。
本文所述的4至6元饱和单杂环被取代时，取代基的位置可处在它们可能的化学位置， 示例性的单杂环的代表性的取代情况如下所示：
Figure PCTCN2017110465-appb-000018
Figure PCTCN2017110465-appb-000019
其中“Sub”表示本文所述的各类取代基；
Figure PCTCN2017110465-appb-000020
表示与其他原子的连接。
如本文所用，“EZH2抑制剂”是指一种能够抑制组蛋白赖氨酸N-甲基转移酶EZH2表达增加的试剂(本发明中是指一种式(I)化合物)，其为PRC2的催化功能亚单位，是负责特定组蛋白H3(H3K27)的Lys27甲基化和对干细胞自我更新不可缺少的。
如本文所用，“由EZH2介导的疾病或病症”是指由于组蛋白赖氨酸N-甲基转移酶EZH2的异常表达而导致异常的表观遗传修饰，从而在患者中形成的异常情况。
如本文所用，“治疗有效量”是指将引起个体的生物学或医学响应，例如降低或抑制酶或蛋白质活性或改善症状、缓解病症、减缓或延迟疾病进程或预防疾病等的本发明化合物的量。
如本文所用，“药学可接受的载体”是指无毒、惰性、固态、半固态的物质或液体灌装机、稀释剂、封装材料或辅助制剂或任何类型辅料，其与患者相兼容，最好为哺乳动物，更优选为人，其适合将活性试剂输送到目标靶点而不终止试剂的活性。
如本文所用，“患者”是指一种动物，最好为哺乳动物，更好的为人。术语“哺乳动物”是指温血脊椎类哺乳动物，包括如猫、狗、兔、熊、狐狸、狼、猴子、鹿、鼠、猪和人类。
如本文所用，“治疗”是指减轻、延缓进展、衰减、预防，或维持现有疾病或病症(例如癌症)。治疗还包括将疾病或病症的一个或多个症状治愈、预防其发展或减轻到某种程度。
制备方法
本发明提供了式(I)化合物的制备方法，本发明中的化合物可以通过多种合成操作制备， 这些化合物的示例性制备方法可以包括(但不限于)下文所述的流程。
较佳地，本发明式(I)化合物可以通过以下方案及实施例中所述的示例性方法以及本领域技术人员所用的相关公开文献操作完成。
在具体操作过程中，可以根据需要对方法中的步骤进行扩展或合并。
方案1:
Figure PCTCN2017110465-appb-000021
步骤1:该硝基化合物转化为相应的胺基化合物，可在酸性条件下用金属(可以是但不限于铁粉，锌粉)进行还原；或者在钯炭催化下加氢还原。
步骤2:吲唑类化合物4位的胺与酮可在酸性条件下，经催化剂(可以是但不限于三乙酰氧基硼氢化钠)催化进行亲核加成。
步骤3:胺与醛的亲核加成反应条件同步骤2。
步骤4:吲唑类化合物芳环上的溴与例如六羰基钼在钯催化剂的作用下发生的插羰反应，再与胺缩合得到目标化合物(I)；所用的钯催化剂可以是但不限于Pd(dppf)Cl2，所用的碱可以是但不限于三乙胺。
方案2:
Figure PCTCN2017110465-appb-000022
步骤1:该硝基化合物转化为相应的胺基化合物，可在酸性条件下用金属(可以是但不限于铁粉，锌粉)进行还原；或者在钯炭催化下加氢还原。
步骤2:吲唑类化合物4位的胺与酮可在酸性条件下，经催化剂(可以是但不限于三乙酰氧基硼氢化钠)催化进行亲核加成。
步骤3:胺与醛的亲核加成反应条件同步骤2。
步骤4:酯的水解，化合物可在如甲醇的碱溶液中水解，所用的碱可以是但不限于氢氧化钠。
步骤5:该吲唑类化合物的酸在缩合剂的作用下与胺缩合成酰胺化合物(I)。
以上各步骤中的反应均是本领域技术人员已知的常规反应。如无特殊说明，合成路线中所使用的试剂和原料化合物均可市购得到，或本领域技术人员根据所设计的不同化合物结构参考已知方法制备得到。
与现有技术相比，本发明的主要优点在于：
提供了一系列结构新颖的4,5,6-三取代吲唑类衍生物，其对EZH2具有高抑制活性，可用作治疗肿瘤的药物。
下面结合具体实施例，进一步阐述本发明。应理解，这些实施例仅用于说明本发明而不用于限制本发明的范围。下列实施例中未注明具体条件的实验方法，通常按照常规条件或按照制造厂商所建议的条件。除非另外说明，否则百分比和份数按重量计算。除非另行定义，本文所用的术语与本领域熟练人员所熟悉的意义相同。此外，任何与所记载内容相似或同等的方法及材料皆可应用于本发明中。
如本文所用，DMB为2,4-二甲氧基苄基，THF为四氢呋喃，EA为乙酸乙酯，PE为石油醚，Ac2O为乙酸酐，NBS为N-溴代琥珀酰亚胺，DCM为二氯甲烷，AIBN为偶氮二异丁腈，Pd(dppf)Cl2为1,1'-双(二苯基磷)二茂铁]二氯化钯，TFA为三氟乙酸，TBSCl为叔丁基二甲基氯硅烷，NCS为N-氯代丁二酰亚胺，DHP为二氢吡喃，LiAlH4为氢化铝锂，PMB为对甲氧基苄基，LiHMDS为二(三甲基硅基)氨基锂，Pd2(dba)3为三(二亚苄基丙酮)二钯，RuPhos为2-二环己基磷-2',6'-二异丙氧基-1,1'-联苯,DMAP为4-二甲氨基吡啶，THP为四氢吡喃，n-BuLi为正丁基锂，TMsOTf为三氟甲磺酸三甲基硅酯，TEBAC为三乙基苄基氯化铵，HATU为2-(7-偶氮苯并三氮唑)-N,N,N',N'-四甲基脲六氟磷酸酯，DMF为二甲基甲酰胺，DMSO为二甲基亚砜，DIEA为N,N-二异丙基乙胺，BINAP为(2R,3S)-2,2'-双二苯膦基-1,1'-联萘。
如本文所用，室温是指约为20-25℃。
中间体1a的制备
Figure PCTCN2017110465-appb-000023
化合物1a-1(22.5g,152mmol)的四氢呋喃(500mL)溶液在冰浴下缓慢加入四氢铝锂(11.5g,0.3mol)，混合物室温搅拌过夜。分别向体系加15mL水，30mL氢氧化钠溶液(15％)，过滤，浓缩滤液得白色固体化合物1a。MS m/z(ESI):N/A。
中间体2a的制备
Figure PCTCN2017110465-appb-000024
步骤1:化合物丙二腈(12g,181.7mmol)的干燥四氢呋喃(225mL)溶液在冰浴下搅拌1小时，分批加入钠氢(4.8g,199.8mmol)并搅拌2小时，逐滴加入化合物2a-1(16.8g,199.8mmol)，混合物缓慢升至室温反应1小时。反应液中加盐酸溶液淬灭，乙酸乙酯萃取，有机层干燥浓缩后得黄色固体化合物2a-2，直接用于下一步。MS m/z(ESI):151[M+H]+。
步骤2:化合物2a-2(28g,181.7mmol)，盐酸(23.2g,4M,636.4mmol)和水(160mL)的混合物搅拌回流5小时。反应液过滤，固体残留物经甲醇重结晶得25g化合物2a-3。MS m/z(ESI):151[M+H]+。
步骤3:化合物2a-3(80mg,0.53mmol)中加入三氯氧磷2mL，混合物在100℃搅拌2小时。LC-MS跟踪至反应完全。混合物冷却后倒入冰水中，用氨水调pH至8，乙酸乙酯萃取 后干燥，浓缩得100mg白色固体化合物2a-4。MS m/z(ESI):169[M+H]+。
步骤4:化合物2a-4(300mg,1.78mmol)，甲醇钠(481mg,8.9mmol)和甲醇(15mL)的混合物在100℃微波16小时。LC-MS跟踪至反应完全。浓缩去除溶剂，残留物加水并调pH至7，过滤得225mg固体化合物2a-5。MS m/z(ESI):419[M+H]+。
步骤5:制备方法同化合物1a，不同的是将1a制法中的化合物1a-1换成化合物2a-5。MS m/z(ESI):152[M+H]+。
中间体3a的制备
Figure PCTCN2017110465-appb-000025
步骤1:化合物3a-1(1g,4.35mmol)的乙醇(10mL)溶液加入氯化铵(697mg,13.04mmol)的水(2mL)溶液和铁粉(364mg,6.52mmol)，混合物搅拌回流2小时。LC-MS跟踪至反应完全。反应液过滤，滤液浓缩的残留物加水溶解，再过滤得700mg白色固体化合物3a-2。MS m/z(ESI):202[M+H]+。
步骤2:化合物3a-2(500mg,45mmol)的硫酸(7.5mL)溶液在-5℃分批加入硝酸钾(278mg,2.75mmol)，混合物在此温度下反应1小时。LC-MS跟踪至反应完全。混合物倒入冰水中，调pH至8，析出的固体过滤，得400mg化合物3a-3。MS m/z(ESI):247[M+H]+。
步骤3:化合物3a-3(2g,8.16mmol)的乙酸(30mL)溶液在40℃下逐滴加入亚硝酸钠(619mg,8.98mmol)的水(5mL)溶液，混合物40℃搅拌1小时。LC-MS跟踪至反应完全。反应液倒入水中，乙酸乙酯萃取，干燥浓缩得2g固体化合物3a-4。MS m/z(ESI):256[M+H]+。
步骤4:制备方法同化合物3a-2，不同的是将3a-2制法中的化合物3a-1换成化合物3a-4，经combiflash纯化得红色固体化合物3a(800mg,45％)。MS m/z(ESI):226[M+H]+。
中间体3a的制备
Figure PCTCN2017110465-appb-000026
步骤1:化合物3a(800mg,3.54mmol)，四氢吡喃酮(708mg,7.08mmol)，与三氟乙酸5mL的1,4-二氧六环(50mL)溶液室温搅拌2小时，加入三乙酰氧基硼氢化钠(2.25mg,10.62mmol)，再室温搅拌4小时。LC-MS跟踪至反应完全。反应液倒入水中，用碳酸氢钠溶液调pH至8，乙酸乙酯萃取后干燥，浓缩得1g化合物4a-1。MS m/z(ESI):312.2[M+H]+。
步骤2:化合物4a-1(1g,3.22mmol)，乙醛(710mg,16.12mmol)，与乙酸3mL的1,4-二氧六环(30mL)溶液室温搅拌2小时，加入三乙酰氧基硼氢化钠(2.05g,9.67mmol)，再室温搅拌过夜。LC-MS跟踪至反应完全。反应液倒入水中，用碳酸氢钠溶液调pH至8，乙酸乙酯萃取后浓缩，combiflash纯化得红色化合物4a(280mg,26％)。MS m/z(ESI):340.1[M+H]+。
中间体5a的制备
Figure PCTCN2017110465-appb-000027
步骤1:化合物5a-1(8g,35.3mmol)的无水甲醇(100mL)溶液中加入二氯亚砜(7.7mL,106.1mmol)，混合物搅拌回流20小时，TLC跟踪至反应结束。反应液冷却后浓缩去掉大部分溶剂，过滤，滤饼减压干燥得7.5g化合物5a-2。MS m/z(ESI):241[M+H]+。
步骤2:化合物5a-2(7.6g,31.64mmol)的乙酸(150mL)溶液分批加入铁粉(14g,253mmol)，将反应温度控制在20-30℃，加完后继续搅拌10min，LC-MS跟踪至反应结束。反应液过滤，滤液倒入水中，乙酸乙酯萃取，有机层浓缩，经combiflash纯化得黄色固体化合物5a-3(3.5g,52.6％)。MS m/z(ESI):211[M+H]+。
步骤3:向化合物5a-3(850mg,4.04mmol)的DMSO(20mL)溶液中逐滴加入NBS(756mg,4.25mmol)的DMSO(1.5mL)溶液，混合物室温搅拌16小时，LC-MS跟踪至反应结束。反应液倒入水中，用乙酸乙酯萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，浓缩，经柱层析纯化得5a-4-1(800mg,68％)和5a-4-2(178mg,15％)，MS m/z(ESI):290.8[M+H]+。
步骤4:制备方法同化合物3a-4，不同的是将3a-4制法中的化合物3a-3换成化合物5a-4-1。MS m/z(ESI):299.8[M+H]+。
步骤5：化合物5a-45(3g,0.01mol)，碘甲烷(4.2g,0.03mol)，碳酸钾(2.2g,0.015mol)的DMF溶液在室温下搅拌2h，LC-MS跟踪至反应结束。将反应液倒入水中(150ml)，搅拌30min，过滤，滤饼减压干燥得化合物5a-6(2.7g,86％)，MS m/z(ESI):316.1[M+H]+。
步骤6：制备方法同化合物3a-4，不同的是将3a-4制法中的化合物3a-3换成化合物5a-6。MS m/z(ESI):284[M+H]+。
步骤67：化合物5a-67(8g,0.026mol)，三氟乙烯基硼酸钾(7g,0.052mol),Pd(dppf)Cl2(0.95g,0.0013mol)，碳酸钠(5.5g,0.052mol)的二氧六环/水(100ml/10ml)混合溶液在100℃下搅拌过夜，LC-MS跟踪至反应结束。反应液冷却至室温后，过滤，滤液用乙酸乙酯萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得米白色固体5a-78(2.7g,40％)，MS m/z(ESI):2632.2[M+H]+。
步骤7：化合物5a-78(1g,4.4mmol)，二氧化铂(100mg,0.22mmol)的乙酸乙酯(20ml)溶液在氢气氛围下室温搅拌3h，LC-MS跟踪至反应结束。反应液过滤，滤液浓缩，经柱层析纯化得黄色固体化合物5a(600mg,59％),MS m/z(ESI):234.2[M+H]+。
中间体6a的制备
Figure PCTCN2017110465-appb-000028
步骤1：向化合物6a-1(2g,102mmol)的二氯甲烷溶液(40ml)中加入三乙胺(3.96g,39.17mmol),TsCl(4.48g,23.50mmol)，反应液在室温下搅拌过夜。将反应液倒入水中，用二氯甲烷萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得油状化合物6a-2(3g, 60％),MS m/z(ESI):257.0[M+H]+。
步骤2：化合物5a-5(2g,6.67mmol)，6a-2(2.56g,10mmol),碳酸钾(1.84g,13.33mmol),碘化钾(1.66g,10mmol)的丙酮溶液回流8h。反应液冷却至室温后，过滤，滤液浓缩，经柱层析纯化得棕色固体化合物6a(600mg,23％),MS m/z(ESI):386.1[M+H]+。
中间体7a-9a的制备
分别以化合物6a和5a-6为原料，参照中间体5a的制备方法，不同的是将步骤6中的三氟乙烯基硼酸钾换成甲基硼酸和环丙基硼酸，碱换成碳酸铯和碳酸钾；将步骤7中的二氧化铂换成钯炭。
Figure PCTCN2017110465-appb-000029
中间体10a的制备
Figure PCTCN2017110465-appb-000030
步骤1：化合物5a-5(20g,0.067mol),NIS(16.6g,0.074mol)的DMF溶液在60℃下搅拌3h，LC-MS跟踪至反应结束。反应液冷却至室温后倒入水中(600ml)，搅拌30min，过滤，滤饼用水洗涤，干燥得化合物10a-1(25g),MS m/z(ESI):425.7[M+H]+。
步骤2：化合物10a-1(25g,0.059mol)，碳酸钾(12.3g,0.09mol)，碘甲烷(12.8g,0.09mol)的DMF(100ml)溶液室温搅拌2h，LC-MS跟踪至反应结束。反应液倒入水中，过滤，滤饼用水洗涤，烘干得化合物10a-2(17g),MS m/z(ESI):439.8[M+H]+。
步骤3：向化合物10a-2(400mg,0.91mmol)和三甲基环三硼氧烷(568mg,4.55mmol),碳酸钾(376mg,2.73mmol)的二氧六环/水(5ml/1ml)的混合溶液中加入催化量的Pd(dppf)Cl2，反应在氮气保护下105℃搅拌16小时，LC-MS跟踪至反应结束。向反应液中加入水，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得化合物10a-3(200mg,83％),MS m/z(ESI):264[M+H]+。
步骤4：同中间体7a中步骤2的制备方法，MS m/z(ESI):234[M+H]+。
中间体11a的制备
Figure PCTCN2017110465-appb-000031
步骤1：在-5-0℃下，向化合物10a-1(16.4g,37.3mmol)的甲醇/冰醋酸(400ml/400ml)的混合溶液中慢慢加入锌粉，在0℃下搅拌20min，然后在室温下再搅拌20min，LC-MS 跟踪至反应结束。反应液用硅藻土过滤，滤液加入水(1L)，用饱和碳酸钠溶液将PH调至7-8。用二氯甲烷萃取，有机相用饱和食盐水洗涤，干燥，过滤，浓缩，经柱层析纯化得灰色固体化合物11a-1(5.3g,50％)，MS m/z(ESI):284.1[M+H]+。
步骤2：化合物11a-1(300mg,1.06mmol)，氰化锌(250mg,2.21mmol)，四三苯基膦钯(128mg,0.11mmol)的DMF(4ml)溶液，在氮气保护下，150℃微波反应30min，LC-MS跟踪至反应结束。向反应液中加入水和乙酸乙酯，分出有机相，用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩。粗品用乙酸乙酯重结晶得灰色固体化合物11a(210mg,64％)，MS m/z(ESI):231.2[M+H]+。
中间体12a的制备
Figure PCTCN2017110465-appb-000032
步骤1：参照5a-7的制备方法，不同的是将三氟乙烯基硼酸钾换成甲基硼酸，将碳酸钠换成碳酸铯。MS m/z(ESI):250.3[M+H]+。
步骤2：向化合物12a-1(672mg,2.7mmol)的醋酸溶液(20ml)中加入选择性氟试剂(2.86g,8.09mmol)，微波160℃加热30min，LC-MS跟踪至反应结束。反应液过滤，滤液浓缩得化合物12a-2(700mg),MS m/z(ESI):268.3[M+H]+。
步骤3：向化合物12a-2(700mg,2.62mmol)的甲醇溶液(25ml)中加入铁粉(440mg,7.86mmol)和醋酸(2ml)，反应回流4h，LC-MS跟踪至反应结束。反应液过滤，滤液浓缩，经柱层析纯化得化合物12a(350mg,56％)，MS m/z(ESI):238.3[M+H]+。
中间体13a的制备
Figure PCTCN2017110465-appb-000033
步骤1：制备方法同化合物3a-4，不同的是将3a-4制法中的化合物3a-3换成化合物5a-4-2。MS m/z(ESI):299.8[M+H]+。
步骤2：制备方法同化合物5a-6，不同的是将5a-6制法中的化合物5a-5换成化合物13a-1。MS m/z(ESI):316.1[M+H]+。
步骤3：向化合物13a-2(110mg,0.35mmol)和三甲基环三硼氧烷(0.15ml,1.05mmol),碳酸铯(228mg,0.7mmol)的二氧六环/水(5ml/0.2ml)的混合溶液中加入催化量的Pd(dppf)Cl2，反应在氮气保护下90℃搅拌10小时，LC-MS跟踪至反应结束。反应液过滤，滤液浓缩，经柱层析纯化得化合物13a-3(70mg,83％),MS m/z(ESI):250.2[M+H]+。
步骤4：向化合物13a-3(70mg,0.28mmol)的甲醇溶液(5ml)中加入钯碳(7mg)，混合物在氢气氛围下室温搅拌10h。反应液过滤，滤液浓缩得化合物13a(55mg,90％),MS m/z(ESI):220.1[M+H]+。
中间体14a的制备
Figure PCTCN2017110465-appb-000034
步骤1：向化合物14a-1(2.5g,7.88mmol)的乙醇溶液(50ml)中加入乙醇钠(1.34g,19.69mmol)，混合物在80℃下搅拌5h，减压蒸馏除去部分乙醇后将剩余物倒入水中，过滤，滤饼干燥得白色固体化合物14a-2(1.9g,90％),MS m/z(ESI):266.5[M+H]+。
步骤2：向化合:14a-2(1.9g,7.11mmol)的甲醇/醋酸(50ml/2.5ml)中加入氢氧化钯(200mg,10％)，混合物在氢气氛围下50℃搅拌6h。反应液冷却至室温后过滤，滤液浓缩得油状化合物14a-3(1.0g)。
步骤3：化合物14a-3(3.2g,12.81mmol)和14a-4(2.0g,12.81mmol)在二氯甲烷/醋酸溶液(100ml/2ml)中室温搅拌30min，在0℃下，加入醋酸硼氢化钠(8.14g,36.42mmol)，混合物在室温下搅拌过夜。将反应液倒入水中，用二氯甲烷萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得油状化合物14a-5(2.1g,68％)，MS m/z(ESI):242.5[M+H]+。
步骤4：向化合物14a-5(2.1g,8.7mmol)的四氢呋喃溶液(20ml)中加入盐酸(7M,20ml)，反应液在70℃搅拌过夜。反应液减压浓缩除去四氢呋喃后，用4M氢氧化钠溶液将PH调至8，用二氯甲烷萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得油状化合物14a(1.4g,81％)，MS m/z(ESI):198.3[M+H]+。
中间体15a-1的制备
Figure PCTCN2017110465-appb-000035
在0℃下，向三氟乙醇(20ml)中加入钠氢(1.13g,28.76mmol),混合物在0℃下继续搅拌30min，向混合物中加入14a-1(3g,9.45mmol),混合物在80℃下搅拌6h。反应液冷却至室温后用二氯甲烷萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得油状化合物15a-1(2.9g,95％),MS m/z(ESI):322.4[M+H]+。
中间体15a-20a的制备
制备方法同中间体14a的制备方法，将14a-3换成相应胺。
Figure PCTCN2017110465-appb-000036
Figure PCTCN2017110465-appb-000037
中间体22a的制备
Figure PCTCN2017110465-appb-000038
步骤1：向化合物14a-4(1.36g,8.71mmol)和化合物22a-1(900mg,7.26mmol)的1,2-二氯乙烷中加入醋酸钠(1.48g,10.9mmol)，反应室温搅拌20min后，加入醋酸硼氢化钠(4.6g,21.8mmol)，反应液室温搅拌过夜，LC-MS跟踪至反应结束。反应液用饱和氢氧化钠水溶液淬灭，并将PH调至8-9，用二氯甲烷萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩得黄色固体化合物22a-2(1.4g,85％)，MS m/z(ESI):228[M+H]+。
步骤2：向化合物22a-2(1.4g,6.2mmol)的水溶液(10ml)中加入三氟乙酸(2ml)，反应室温搅拌过夜，LC-MS跟踪至反应结束。向反应液中加入饱和碳酸氢钠溶液至PH＝8-9，用二氯甲烷萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得黄色油状化合物22a(500mg,44％)，MS m/z(ESI):184[M+H]+。
化合物23a的制备
Figure PCTCN2017110465-appb-000039
步骤1：化合物14a-4(1g,6.4mmol)，化合物23a-1(0.83g,6.4mmol)和DIEA(0.82g,6.4mmol)的甲醇溶液(20ml)在室温下搅拌1h，加入醋酸硼氢化钠(2.7g,12.8mmol),室温搅拌过夜，LC-MS跟踪至反应结束。用饱和碳酸氢钠淬灭反应，并将PH调至8-9，反应液用二氯甲烷萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩得化合物23a-2(1.1g),MS m/z(ESI):234[M+H]+。
步骤2：化合物23a-2(1.1g,4.7mmol)的盐酸溶液(5M,20ml)室温搅拌16h，TLC跟踪至反应结束。向反应液中加入饱和碳酸氢钠溶液至PH＝8-9，用二氯甲烷萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩得化合物23a(300mg)。
中间体24a的制备
Figure PCTCN2017110465-appb-000040
步骤1：向化合物24a-1(8.6g,50.5mmol)的甲苯溶液中加入乙二醇(3.86ml,75.8mmol)和对甲苯磺酸(170mg,1.01mmol),混合物回流除水10h。将反应液浓缩，经柱层析纯化得化合物24a-2(5.5g,51％)。
步骤2：在-78℃下，向化合物24a-2(4.05g,18.9mmol)的四氢呋喃溶液中逐滴加入LDA(2M,14.1ml)，在该温度下搅拌30min后加入碘甲烷(2.35ml,37.8mmol)，反应液自然升至室温，再搅拌2h，TLC跟踪至反应结束。将反应用水淬灭后用乙酸乙酯萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得化合物24a-3(3.5g,72％)。
步骤3：将氢氧化钠(1.58g,39.4mmol)加入到化合物24a-3(3.0g,13.1mmol)的乙醇/水(20ml/20ml)溶液中，混合物在60℃下搅拌10h，TLC跟踪至反应结束。反应液冷却至室温后，用1M的盐酸将PH调至3，用乙酸乙酯萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩得化合物24a-4(2.5g,95％)。
步骤4：向化合物24a-4(2.2g,11mmol)的二氧六环溶液中加入盐酸(4M,30ml)，混合物在65℃下搅拌3h。反应液冷却至室温后，用乙酸乙酯萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩得化合物24a-5(2.0g,100％)。
步骤5：向化合物24a-5(1g,6.41mmol)的甲苯溶液中加入DPPA(1.66ml,7.69mmol)，三乙胺(1.33ml,9.62mmol)和苯甲醇(0.8ml,7.69mmol),反应液回流10h。将反应液浓缩得到化合物24a(700mg,42％),MS m/z(ESI):262.1[M+H]+。
实施例1：4-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-1)的制备
Figure PCTCN2017110465-appb-000041
步骤1:化合物4a(100mg,0.296mmol)的N,N-二甲基甲酰胺5mL溶液在冰浴下加入钠氢(24mg,0.591mmol)，冰浴下搅拌30分钟，加入碘甲烷(84mg,0.591mmol)，混合物再搅拌2小时。LC-MS跟踪至反应完全。反应液倒入水中，乙酸乙酯萃取后干燥，浓缩得60mg化合物1-1。MS m/z(ESI):354.2[M+H]+。
步骤2:化合物1-1(50mg,0.14mmol)，化合物2a(50mg,0.21mmol)，C6MoO6(50mg,0.21mmol)，Pd(dppf)Cl2(10mg,0.01mmol)和三乙胺(60mg,0.57mmol)的N,N-二甲基甲酰胺3mL溶液在氩气氛围下140℃微波反应40分钟。LC-MS跟踪至反应完全。反应液倒入水中，乙酸乙酯萃取后浓缩，经Prep-HPLC纯化得白色固体化合物p-1(7.5mg,11.3％)。MS m/z(ESI):468.3[M+H]+；1H NMR(400MHz,DMSO):δ11.41(s,1H),8.05(d,1H),7.90(t,1H),7.28(s,1H),6.07(s,1H),4.22(d,2H),3.94(s,3H),3.88-3.65(m,5H),3.28-3.13(m,5H),2.27(s,3H),2.14(s,3H),1.93-1.77(m,1H),1.58-1.24(m,3H),0.74(t,3H).
实施例2：4-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-甲基-1H-吲唑-6-甲酰胺(化合物p-2)的制备
Figure PCTCN2017110465-appb-000042
制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1换成化合物4a。经Prep-HPLC纯化得白色固体化合物p-2(5mg,3.7％)。MS m/z(ESI):454.3[M+H]+；1H NMR(400MHz,DMSO-6)δ12.99(s,1H),11.40(s,1H),8.09(s,1H),7.93(t,1H),7.14(s,1H),6.07(s,1H),4.20(d,2H),3.82-3.65(m,5H),3.29-3.14(m,5H),2.27(s,3H),2.14(s,3H),1.88-1.77(m,1H),1.57-1.46(m,1H),1.39-1.25(m,2H),0.74(t,3H).
实施例3：7-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-6-甲基-1H-咪唑-5-甲酰胺(化合物p-3)的制备
Figure PCTCN2017110465-appb-000043
步骤1:化合物3-1(20g,0.16mol)的二氯甲烷200mL溶液在氩气氛围下于0℃加入乙酸酐(17.7g,0.17mol)和三乙胺(18.4g,0.18mol)，混合物室温下搅拌2小时。反应液中加水，分离有机层，干燥浓缩得25.4g白色固体化合物3-2。MS m/z(ESI):164.1[M+H]+。
步骤2:化合物3-2(25.3g,0.15mol)的乙酸250mL溶液在10℃逐滴加入Br2(25.2g,0.15mol)的乙酸50mL溶液，混合物在55℃搅拌16小时。反应液过滤，滤液中加水，再过滤，两次的滤饼合并后用乙醇洗涤，滤饼在低于50℃时减压干燥，得32.0g黄色固体化合物3-3。MS m/z(ESI):242[M+H]+。
步骤3:化合物3-3(32g,0.13mol)的硫酸(13.02g,0.13mol)溶液在氩气氛围下于冰浴加入硝酸(14.54g,0.15mol,61％)，混合物在冰浴下继续搅拌2小时。LC-MS跟踪至反应完全。反应液倒入冰水中，过滤，滤饼用水洗涤，并在低于50℃时减压干燥，得40.0g固体化合物3-4。MS m/z(ESI):287[M+H]+。
步骤4:化合物3-4(16g,35mmol)的硫酸(170mL,50％)溶液在100℃搅拌6小时。LC-MS跟踪至反应完全。反应液中加水，过滤，滤饼用乙醇洗涤，并在低于50℃时减压干燥，得5.0g固体化合物3-5。MS m/z(ESI):245[M+H]+。
步骤5:制备方法同化合物3a-4，不同的是将3a-4制法中的化合物3a-3换成化合物3-5。MS m/z(ESI):256[M+H]+。
步骤6:制备方法同化合物3a-2，不同的是将3a-2制法中的化合物3a-1换成化合物3-6。MS m/z(ESI):228[M+H]+。
步骤7:制备方法同化合物4a-1，不同的是将4a-1制法中的化合物3a换成化合物3-7。 MS m/z(ESI):312[M+H]+。
步骤8:制备方法同化合物4a，不同的是将4a制法中的化合物4a-1换成化合物3-8。MS m/z(ESI):312[M+H]+。
步骤9:制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1换成化合物3-9。经Prep-HPLC纯化得白色固体化合物p-3(4mg,2％)。MS m/z(ESI):454.3[M+H]+；1H NMR(400MHz,DMSO-6)δ12.81(s,1H),11.39(s,1H),8.41(s,1H),8.00(s,1H),7.83(t,1H),7.43(s,1H),6.07(s,1H),4.20(d,2H),3.93-3.61(m,5H),3.29-3.10(m,5H),2.37(s,3H),2.15(s,3H),1.83-1.80(m,1H),1.47-1.44(m,1H),1.29-1.23(m,2H),0.74(t,3H).
实施例4：7-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-1,6二甲基-1H-咪唑-5-甲酰胺(化合物p-4)的制备
Figure PCTCN2017110465-appb-000044
步骤1:化合物3-9(150mg,0.443mmol)的DMF(10mL)溶液加入碳酸钾(122mg,0.886mmol)和碘甲烷(125mg,0.886mmol)，混合物室温搅拌过夜。LC-MS跟踪至反应结束。反应液倒入水中，乙酸乙酯萃取，有机层浓缩并经combiflash纯化得化合物4-1(80mg,51％)。MS m/z(ESI):354.2[M+H]+。
步骤2:制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1换成化合物4-1。经Prep-HPLC纯化得白色固体化合物p-4(2mg,2％)。MS m/z(ESI):468.3[M+H]+；1H NMR(400MHz,DMSO-6)δ11.39(s,1H),8.25(s,1H),7.80(t,1H),7.36(s,1H),6.07(s,1H),4.19(d,2H),4.08(s,3H),3.92-3.59(m,6H),3.53-3.46(m,2H),3.13-3.00(m,2H),2.32(s,3H),2.15(s,3H),1.54-1.43(m,1H),1.33-1.02(m,3H),0.70(t,3H).
实施例5：1-(二氟甲基)-4-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-甲基-1H-吲唑-6-甲酰胺(化合物p-5)的制备
实施例6：2-(二氟甲基)-4-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-甲基-2H-吲唑-6-甲酰胺(化合物p-6)的制备
Figure PCTCN2017110465-appb-000045
步骤1:制备方法同化合物1-1，不同的是将1-1制法中的化合物碘甲烷换成二氟氯乙酸钠，得到化合物5-1与5-2的混合物。MS m/z(ESI):388.1[M+H]+。
步骤2:制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1换成化合物5-1与5-2的混合物。经Prep-HPLC纯化得白色固体化合物p-5(10mg,6.4％).1H NMR(400MHz,DMSO-6)δ11.40(s,1H),8.91(s,1H),8.16-7.86(m,2H),7.31(s,1H),6.07(s,1H),4.21(d,2H),3.86-3.66(m,5H),3.24-3.10(m,5H),2.26(s,3H),2.15(s,3H),1.92-1.75(m,1H),1.56-.124(m,3H),0.74(t,3H).化合物p-6(12mg,7.7％；1H NMR(400 MHz,DMSO-6)δ11.40(s,1H),8.48(s,1H),8.12(t,1H),8.07(t,1H),7.44(s,1H),6.06(s,1H),4.22(d,2H),3.81-3.72(m,5H),3.25-3.05(m,5H),2.28(s,3H),2.14(s,3H),1.87-1.70(m,1H),1.54-1.27(m,3H),0.76(t,3H).MS m/z(ESI):504[M+H]+。
实施例50：4-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-2,5二甲基-2H-吲唑-6-甲酰胺(化合物p-50)的制备
Figure PCTCN2017110465-appb-000046
制备方法同P-6，其中步骤1参见化合物1-1的方法制备。MS m/z(ESI):468.2[M+H]+。
实施例7：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(乙基(四氢-2H-吡喃-4-基)氨基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-7)的制备
Figure PCTCN2017110465-appb-000047
制备方法同化合物p-1，不同的是将p-1制法中的化合物2a换成化合物1a。经Prep-HPLC纯化得白色固体化合物p-7(9mg,7.8％)。MS m/z(ESI):452[M+H]+；1H NMR(400MHz,DMSO-6)δ11.44(s,1H),8.10(t,1H),8.06(s,1H),7.29(s,1H),5.84(s,1H),4.27(d,2H),3.95(s,3H),3.85-3.65(m,2H),3.28-3.00(m,5H),2.26(s,3H),2.20(s,3H),2.08(s,3H),1.93-1.73(m,1H),1.56-1.24(m,3H),0.74(t,3H).
实施例8：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(乙基(1-(2,2,2-三氟乙基)哌啶-4-基)氨基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-8)的制备
Figure PCTCN2017110465-appb-000048
步骤1:制备方法同化合物4a-1，不同的是将4a-1制法中的四氢吡喃酮换成化合物8.1。MS m/z(ESI):390[M+H]+。
步骤2:制备方法同化合物4a，不同的是将4a制法中的化合物4a-1换成化合物8-1。MS m/z(ESI):418[M+H]+。
步骤3:制备方法同化合物1-1，不同的是将1-1制法中的化合物4a换成化合物8-2。MS m/z(ESI):434.9[M+H]+。
步骤4:制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1与2a换成化合物 8-3&8-4与1a。经Prep-HPLC纯化得白色固体化合物p-8(8mg,8％)。MS m/z(ESI):533[M+H]+；1H NMR(400MHz,DMSO-6)δ11.47(s,1H),8.13(t,1H),8.09(s,1H),7.33(s,1H),5.85(s,1H),4.29(d,2H),3.99-3.94(m,5H),3.43-3.11(m,5H),3.05-2.91(m,2H),2.27(s,3H),2.21(s,3H),2.09(s,3H),1.74-1.57(m,2H),1.52-1.36(m,2H),0.75(t,3H).
实施例9：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(乙基(4-((2-甲氧基乙基)(甲基)氨基)环己基)氨基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-9)的制备
Figure PCTCN2017110465-appb-000049
步骤1:制备方法同化合物4a-1，不同的是将4a-1制法中的四氢吡喃酮换成化合物9.1。MS m/z(ESI):390[M+H]+。
步骤2:制备方法同化合物4a，不同的是将4a制法中的化合物4a-1换成化合物9-1。MS m/z(ESI):418[M+H]+。
步骤3:制备方法同化合物1-1，不同的是将1-1制法中的化合物4a换成化合物9-2。MS m/z(ESI):434.9[M+H]+。
步骤4:制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1与2a换成化合物9-3与1a。经Prep-HPLC纯化得白色固体化合物p-9(4mg,3.6％)。MS m/z(ESI):537.1[M+H]+；1H NMR(400MHz,DMSO-6)δ11.44(s,1H),8.09(t,1H),8.02(s,1H),7.26(s,1H),5.84(s,1H),4.28(d,2H),3.95(s,3H),3.29-3.27(m,4H),3.17(s,3H),2.94-2.85(m,1H),2.45-2.41(m,2H),2.24(s,3H),2.20(s,3H),2.10(s,3H),2.08(s,3H),1.70-1.54(m,3H),1.35-1.05(m,5H),0.73(t,3H).
实施例10：1-(二氟甲基)-N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(乙基(四氢-2H-吡喃-4-基)氨基)-5-甲基-1H-吲唑-6-甲酰胺(化合物p-10)的制备
Figure PCTCN2017110465-appb-000050
制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1与2a换成化合物5-1与1a。经Prep-HPLC纯化得白色固体化合物p-10(16mg,16％)。MS m/z(ESI):488[M+H]+；1H NMR(400MHz,DMSO-6)δ11.47(s,1H),8.50(s,1H),8.31(t,1H),8.14(t,1H),7.46(s,1H),5.85(s,1H),4.28(d,2H),3.85-3.69(m,2H),3.39-3.11(m,5H),2.28(s,3H),2.20(s,3H),2.09(s,3H),1.59-1.22(s,4H),0.77(t,3H).
实施例11：4-(乙基(1-(2,2,2-三氟乙基)哌啶-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基基)甲基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-11)的制备
实施例12：4-(乙基(1-(2,2,2-三氟乙基)哌啶-4-基)氨基)-N-((4-甲氧基-6-甲 基-2-氧代-1,2-二氢吡啶-3-基基)甲基)-2,5-二甲基-2H-吲唑-6-甲酰胺(化合物p-12)的制备
Figure PCTCN2017110465-appb-000051
制备方法同化合物p-1，不同的是将p-1制法中的化合物1-1换成化合物8-3与8-4的混合物。经Prep-HPLC纯化得白色固体化合物p-11(9mg,12％)；1H NMR(400MHz,DMSO-6)δ11.41(s,1H),8.04(s,1H),7.90(t,1H),7.28(s,1H),6.07(s,1H),4.23(d,2H),3.95(s,3H),3.79(s,3H),3.29-2.51(m,9H),2.27(s,3H),2.15(s,3H),1.90-1.73(m,1H),1.56-1.31(m,3H),0.74(t,3H).p-12(3mg,4％)；1H NMR(400MHz,DMSO-6)δ11.40(s,1H),8.39(s,1H),7.89(t,1H),7.22(s,1H),6.07(s,1H),4.20(d,2H),4.09(s,3H),3.79(s,3H),3.24-2.64(m,9H),2.25(s,3H),2.15(s,3H),1.92-1.83(m,1H),1.62-1.50(m,1H),1.38-1.21(m,2H),0.72(t,3H).MS m/z(ESI):549[M+H]+。
实施例13：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(((1S，4S)-4-(二甲基氨基)环己基)(乙基)氨基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-13)的制备
实施例14：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(((1R，4R)-4-(二甲基氨基)环己基)(乙基)氨基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-14)的制备
Figure PCTCN2017110465-appb-000052
步骤1:化合物13-1(8g,35.3mmol)的无水甲醇(100mL)溶液中加入二氯亚砜(7.7mL,106.1mmol)，混合物室温搅拌回流20小时。TLC跟踪至反应结束。反应液冷却后浓缩去掉大部分溶剂，过滤，滤饼减压干燥得7.5g化合物13-2。MS m/z(ESI):241[M+H]+。
步骤2:化合物13-2(7.6g,31.64mmol)的乙酸(150mL)溶液分批加入铁粉(14g,253mmol)，加完后反应液过滤，滤液倒入水中，乙酸乙酯萃取，有机层浓缩，经combiflash纯化得黄色固体化合物13-3(3.5g,52.6％)。MS m/z(ESI):211[M+H]+。
步骤3:化合物13-3(1g,4.76mmol)的DMSO(20mL)溶液逐滴加入NBS(931mg,5.23mmol) 的DMSO(1.5mL)溶液，混合物室温搅拌2小时。LC-MS跟踪至反应结束。反应液倒入水中，过滤，滤饼减压干燥得700mg黄色固体化合物13-4。MS m/z(ESI):290.8[M+H]+。
步骤4:制备方法同化合物3a-4，不同的是将3a-4制法中的化合物3a-3换成化合物13-4。MS m/z(ESI):299.8[M+H]+。
步骤5:制备方法同化合物1-1，不同的是将1-1制法中的化合物4a换成化合物13-5。MS m/z(ESI):315.8[M+H]+。
步骤6:化合物13-6(900mg,2.87mmol)，甲基硼酸(343mg,5.73mmol)，Pd(dppf)Cl2(210mg,0.286mmol)，碳酸钠(607mg,5.73mmol)，1,4-二氧六环(20mL)和2mL水的混合物在氩气氛围下100℃微波反应8小时。LC-MS跟踪至反应完全。反应液倒入水中，乙酸乙酯萃取浓缩后经combiflash纯化得黄色固体化合物13-7(300mg,20％)。MS m/z(ESI):249.9[M+H]+。
步骤7:制备方法同化合物13-3，不同的是将13-3制法中的化合物13-2换成化合物13-7，混合物加热至90℃搅拌7小时。MS m/z(ESI):220[M+H]+。
步骤8:制备方法同化合物4a-1，不同的是将4a-1制法中的四氢吡喃酮换成4-二甲氨基环己酮。MS m/z(ESI):345.1[M+H]+。
步骤9:制备方法同化合物4a，不同的是将4a制法中的4a-1换成化合物13-9。MS m/z(ESI):373.1[M+H]+。
步骤10:化合物13-10(270mg,0.724mmol)的THF(10mL)溶液中加入甲醇(4mL)和氢氧化钠(4mL,3M)，混合物室温搅拌过夜。LC-MS跟踪至反应结束。反应液调pH至6，乙酸乙酯萃取除杂，水层浓缩，残留物经DCM:MeOH＝10:1洗涤，过滤后浓缩滤液，得200mg化合物13-11。MS m/z(ESI):357.3[M+H]+。
步骤11:化合物13-11(80mg,0.223mmol)的DMF(3mL)溶液中加入HATU(127mg,0.335mmol)，DIPEA(115mg,0.892mmol)和化合物1a(84mg,0.446mmol)，混合物在室温下搅拌过夜。LC-MS跟踪至反应结束。反应液经乙酸乙酯/水体系萃取，有机层浓缩，combiflash纯化得白色固体化合物p-13(3mg,2.7％)；1H NMR(400MHz,DMSO)δ11.46(s,1H),8.12(t,1H),8.03(s,1H),7.26(s,1H),5.83(s,1H)，4.26(d,2H),3.93(s,3H)，3.57(s,1H),3.18-2.98(m,3H),2.28-2.18(m,12H),2.07(s,3H),1.71(s,2H),1.41-1.19(m,6H),0.77(t,3H)和p-14(9mg,8％)。MS m/z(ESI):493.4[M+H]+；1H NMR(400MHz,DMSO-6)δ11.44(s,1H),8.09(t,1H),8.02(s,1H),7.26(s,1H),5.84(s,1H),4.28(d,2H),3.95(s,3H),3.28-3.03(m,2H),2.95-2.84(m,2H),2.28(s,3H),2.20(s,3H),2.15-1.99(m,9H),1.74-1.64(m,4H),1.34-1.20(m,2H),1.12-0.99(m,2H),0.73(t,3H).
实施例15：4-(乙基(4-((2-甲氧基乙基)(甲基)氨基)环己基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-1,5-二甲基-1H-吲唑-6-甲酰胺(化合物p-15)的制备
Figure PCTCN2017110465-appb-000053
步骤1:制备方法同化合物4a-1，不同的是将4a-1制法中的四氢吡喃酮换成化合物15.1。MS m/z(ESI):389.1[M+H]+。
步骤2:制备方法同化合物4a，不同的是将4a制法中的化合物4a-1换成化合物15-1。MS m/z(ESI):417.4[M+H]+。
步骤3:制备方法同化合物13-11，不同的是将13-11制法中的化合物13-10换成化合物15-2。MS m/z(ESI):403.4[M+H]+。
步骤4:制备方法同化合物p-13，不同的是将p-13制法中的化合物13-11换成化合物15-3。经Prep-HPLC纯化得白色固体化合物p-15(25mg,10％)。MS m/z(ESI):553.3[M+H]+；1H NMR(400MHz,DMSO-6)δ11.39(s,1H),8.02(s,1H),7.88(t,1H),7.25(s,1H),6.07(s,1H),4.23(d,2H),3.97(s,3H),3.79(s,3H),3.51-3.41(m,2H),3.29-3.27(m,2H),3.20-3.15(m,4H),2.94-2.84(m,1H),2.46-2.43(m,2H),2.24(s,3H),2.15(s,3H),2.10(s,3H),1.72-1.57(m,3H),1.33-1.07(m,5H),0.73(t,3H).
实施例16：5-乙基-4-(乙基(四氢-2H-吡喃-4-基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-1-甲基-1H-吲唑-6-甲酰胺(化合物p-16)的制备
Figure PCTCN2017110465-appb-000054
步骤1:制备方法同化合物13-7，不同的是将13-7制法中的甲基硼酸换成化合物16.1。MS m/z(ESI):262.1[M+H]+。
步骤2:化合物16-1(50mg,0.2mmol)的甲醇(5mL)和1mL四氢呋喃溶液加入钯炭(15mg)，混合物在氢气氛围下室温搅拌过夜。LC-MS跟踪至反应完全。反应液经过滤，浓缩得化合物16-2。MS m/z(ESI):234.2[M+H]+。
步骤3:制备方法同化合物4a-1，不同的是将4a-1制法中的化合物3a换成化合物16-2。MS m/z(ESI):318.2[M+H]+。
步骤4:制备方法同化合物4a，不同的是将4a制法中的化合物4a-1换成化合物16-3。MS m/z(ESI):346.2[M+H]+。
步骤5:制备方法同化合物13-11，不同的是将13-11制法中的化合物13-10换成化合物16-4。MS m/z(ESI):332.2[M+H]+。
步骤6:制备方法同化合物p-13，不同的是将p-13制法中的化合物13-11换成化合物16-5。经Prep-HPLC纯化得白色固体化合物p-16(46mg,16％)。MS m/z(ESI):482.2[M+H]+；1H NMR(400MHz,DMSO)δ11.41(s,1H),8.07(s,1H),7.92(t,1H),7.31(s,1H)，6.07(s,1H)，4.26(d,2H)，3.95(s,3H)，3.83-3.71(m,2H)，3.78(s,3H),3.24-3.18(m,5H)，2.96-2.92(m,2H)，2.17(s,3H)，1.85(s,1H)，1.55(s,1H)，1.38-1.29(m,2H),0.99(t,3H),0.77(t,3H).
实施例17：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-乙基-4-(乙基(四氢-2H-吡喃-4-基)氨基)-1甲基-1H-吲唑-6-甲酰胺(化合物p-17)的制备
Figure PCTCN2017110465-appb-000055
制备方法同化合物p-13，不同的是将p-13制法中的化合物13-11与2a换成化合物16-5与1a。经Prep-HPLC纯化得白色固体化合物p-17(52mg,19％)。MS m/z(ESI):466.2[M+H]+；1H NMR(400MHz,DMSO)δ11.44(s,1H),8.10(t,1H),8.07(s,1H),7.30(s,1H)，5.84(s,1H)，4.27(d,2H)，3.93(s,3H)，3.82-3.70(m,2H)，3.23-3.12(m,5H)，2.95-2.92(m,2H)，2.21(s,3H)，2.08(s,3H)，1.88(s,1H)，1.52(s,1H)，1.32-1.27(m,2H),0.94(t,3H),0.77(t,3H).
实施例18：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-乙基-4-(乙基((1S,4S)-4-(3-甲基氮杂环丁烷嗪-1-基)环己基)氨基)-1-甲基-1H-吲唑-6-甲酰胺(化合物p-18)的制备
实施例19：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-乙基-4-(乙基((1R,4R)-4-(3-甲基氮杂环丁烷嗪-1-基)环己基)氨基)-1-甲基-1H-吲唑-6-甲酰胺(化合物p-19)的制备
Figure PCTCN2017110465-appb-000056
步骤1:制备方法同化合物4a-1，不同的是将4a-1制法中的化合物3a与四氢吡喃酮换成化合物16-2与18.1。MS m/z(ESI):401[M+H]+。
步骤2:制备方法同化合物4a，不同的是将4a制法中的化合物4a-1换成化合物18-1。MS m/z(ESI):429.3[M+H]+。
步骤3:制备方法同化合物13-11，不同的是将13-11制法中的化合物13-10换成化合物18-2。MS m/z(ESI):415[M+H]+。
步骤4:制备方法同化合物p-13，不同的是将p-13制法中的化合物13-11与2a换成化合物18-3与1a。经Prep-HPLC纯化得白色固体化合物p-18；1H NMR(400MHz,DMSO)δ11.46(s,1H),8.13(t,1H),8.05(s,1H),7.30(s,1H),5.87(s,1H),4.30(d,2H),3.97(s,3H),3.86-3.82(m,1H),3.38(t,2H),3.11(s,3H),2.93(t,4H),2.67(t,2H),2.36-2.29(m,1H),2.23(s,3H),2.11(s,3H),1.89-1.55(m,6H),0.96(t,3H),0.77 (t,6H).与p-19；1H NMR(400MHz,DMSO)δ11.46(s,1H),8.13(t,1H),8.05(s,1H),7.29(s,1H),5.87(s,1H),4.30(d,2H),3.97(s,3H),3.93-3.89(m,1H),3.43(s,2H),3.13(s,3H),3.08-2.90(m,4H),2.64-2.59(m,2H),2.24(s,3H),2.11(s,3H),2.08(s,1H),1.76-1.43(m,6H),1.20(s,3H),0.97(t,3H),0.79(t,3H).。MS m/z(ESI):549[M+H]+。
实施例20：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-乙基-4-(乙基((1S，4S)-4-((2-甲氧基乙基)(甲基)氨基)环己基)氨基)-1-甲基-1H-吲唑-6-甲酰胺(化合物p-20)的制备
Figure PCTCN2017110465-appb-000057
步骤1:制备方法同化合物4a-1，不同的是将4a-1制法中的化合物3a与四氢吡喃酮换成化合物16-2与20.1。MS m/z(ESI):403.3[M+H]+。
步骤2:制备方法同化合物4a，不同的是将4a制法中的化合物4a-1换成化合物20-1。MS m/z(ESI):431.3[M+H]+。
步骤3:制备方法同化合物13-11，不同的是将13-11制法中的化合物13-10换成化合物20-2。MS m/z(ESI):417.4[M+H]+。
步骤4:制备方法同化合物p-13，不同的是将p-13制法中的化合物13-11与2a换成化合物20-3与1a。经Prep-HPLC纯化得白色固体化合物p-20；1H NMR(400MHz,DMSO-6)δ11.43(s,1H),8.10(t,1H),8.05(s,1H),7.30(s,1H),5.84(s,1H),4.28(d,2H),3.96(s,3H),3.54-3.47(m,2H),3.25(s,3H),3.21-2.77(m,8H),2.67-2.50(m,3H),2.20(s,3H),2.08(s,3H),1.98-1.71(m,3H),1.62-1.52(m,1H),1.44-1.21(m,4H),0.95(t,3H),0.76(t,3H).MS m/z(ESI):551.3[M+H]+。
实施例21：5-乙基-4-(甲基((1R，4R)-4-((2-甲氧基乙基)(甲基)氨基)环己基)氨基)-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-1-甲基-1H-吲唑-6-甲酰胺(化合物p-21)的制备
Figure PCTCN2017110465-appb-000058
制备方法同化合物p-13，不同的是将p-13制法中的化合物13-11换成化合物20-3。经Prep-HPLC纯化得白色固体化合物p-21。MS m/z(ESI):567[M+H]+；1H NMR(400MHz,DMSO-6)δ11.42(s,1H),9.12(s,1H),8.04(s,1H),7.94(t,1H),7.29(s,1H),6.07(s,1H),4.22(d,2H),3.94(s,3H),3.77(s,3H),3.60-3.54(m,2H),3.33-3.20(m,5H),3.15-2.86(m,6H),2.63(d,3H),2.14(s,3H),1.97-1.82(m,3H),1.63-1.54(m,1H),1.45-1.27(m,4H),0.98(t,3H),0.75(t,3H)。
实施例22：4-((4-(二甲基氨基)环己基)(乙基)氨基)-5-乙基-N-((4-甲氧基-6-甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)1-甲基-1H-吲唑-6-甲酰胺(P-22)的制备
Figure PCTCN2017110465-appb-000059
步骤1：化合物5a(100mg,0.408mmol),4-二甲基环己酮(121mg,0.857mmol)的二氧六环/三氟乙酸(10ml/2ml)混合溶液在室温下搅拌2h，加入醋酸硼氢化钠(272mg,1.29mmol)，混合物在室温下继续搅拌1h，LC-MS跟踪至反应结束。反应用饱和碳酸氢钠溶液淬灭，并将PH调至8，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，过滤，浓缩得棕色油状化合物22-1(150mg),MS m/z(ESI):359.3[M+H]+。
步骤2：化合物22-1(150mg,0.418mmol)，乙醛(92mg,2.07mmol)的二氧六环/乙酸(20ml/2ml)的混合溶液在室温下搅拌1h，加入醋酸硼氢化钠(443mg,2.09mmol)，混合物在室温下搅拌1h，LC-MS跟踪至反应结束。反应用饱和碳酸氢钠溶液淬灭，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，过滤，浓缩得棕色油状化合物22-2(150mg),MS m/z(ESI):387.4[M+H]+。
步骤3：向化合物22-2(150mg,0.388mmol)的甲醇溶液中加入氢氧化钠溶液(4M,2ml)，混合物在50℃下搅拌5h，LC-MS跟踪至反应结束。反应液减压浓缩除去甲醇后加水稀释，用乙酸乙酯萃取除去杂质，水相用盐酸(3M)将PH调至4，用乙酸乙酯萃取，有机相用无水硫酸钠干燥，过滤，浓缩得黑色油状物22-3(150mg)，MS m/z(ESI):373.4[M+H]+。
步骤4：向化合物22-3(140mg,0.576mmol)的DMF溶液中加入化合物2a(106mg,0.564mmol)，HATU(214mg,0.564mmol),DIPEA(194mg,1.50mmol)，混合物在室温下搅拌5h，LC-MS跟踪至反应结束。将反应液倒入水中，用乙酸乙酯萃取，有机相用食盐水洗涤，无水硫酸钠干燥，过滤，浓缩，经Pre-HPLC纯化得白色固体化合物P-22(6mg,3％),MS m/z(ESI):523.3[M+H]+。1H NMR(400MHz,DMSO)δ11.40(s,1H),8.22(s,1H),8.03(s,1H),7.92(t,1H),7.26(s,1H),6.06(s,1H),4.22(d,J＝4.4Hz,2H),3.99(s,3H),3.94(s,3H),3.18-3.05(m,2H),2.93-2.88(m,4H),2.16(s,6H),2.14(s,3H),1.72-1.52(m,4H),1.34-1.11(m,4H),0.98(t,J＝7.6Hz,3H),0.75(t,J＝7.2Hz,3H).
Figure PCTCN2017110465-appb-000060
实施例的目标化合物如式(II)所示，取代基R1，R11，R12和Y如下表所示。
通用步骤:化合物P-23至P-42以化合物5a-12a和14a-21a，23a或相应的酮为原料，将步骤4中2a换成1a，参照实施例22的类似方法进行制备。(Et代表乙基，Me代表甲基)
Figure PCTCN2017110465-appb-000061
Figure PCTCN2017110465-appb-000062
实施例44：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(乙基(4-(甲基(2,2,2-三氟乙基)氨基)环己基)氨基)-1,5-二甲基-1H-吲唑-6-甲酰胺(P-44)的制备
Figure PCTCN2017110465-appb-000063
步骤1：制备方法同化合物22-1，不同的是将化合物5a和4-二甲基环己酮换成化合物8a和14a-4，反应温度为0℃。MS m/z(ESI):360.2[M+H]+。
步骤2：制备方法同化合物22-2，不同的是将化合物22-1换成44-1，反应温度为0℃。MS m/z(ESI):388.3[M+H]+。
步骤3：化合物44-2(750mg,1.94mmol)的三氟乙酸/水(10ml/1ml)溶液在50℃下搅拌5h。反应液冷却至室温后，加入饱和碳酸氢钠水溶液淬灭反应，用二氯甲烷萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得白色固体化合物44-3(650mg,97％)，MS m/z(ESI):344.3[M+H]+。
步骤4：化合物44-3(500mg,1.46mmol)和化合物44.1(288mg,2.91mmol)的二氯甲烷/醋酸溶液(10ml/0.2ml)在0℃下搅拌1h，在0℃下加入醋酸硼氢化钠(925mg,4.37mmol),混合物在0℃下继续搅拌2h，LC-MS跟踪至反应结束。用饱和碳酸氢钠水溶液将反应淬灭， 用二氯甲烷萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得油状化合物44-4(130mg,26％),MS m/z(ESI):427.3[M+H]+。
步骤5：制备方法同化合物44-4，不同的是将化合物44-3和44.1换成化合物44-4和甲醛，溶剂换成甲醇/醋酸溶液。MS m/z(ESI):441.3[M+H]+。
步骤6：制备方法同化合物22-3，不同的是将化合物22-2换成44-5。MS m/z(ESI):427.3[M+H]+。
步骤7：制备方法同化合物P-22，不同的是将化合物22-3和2a换成化合物44-6和1a。经Pre-HPLC纯化得白色固体化合物P-44(54mg,37％),MS m/z(ESI):561.3[M+H]+。1H NMR(500MHz,DMSO)δ11.48(s,1H),8.16(t,J＝4.8Hz,1H),8.07(s,1H),7.31(s,1H),5.87(s,1H),4.30(d,J＝4.8Hz,2H),3.98(s,3H),3.76-3.72(m,1H),3.27-3.08(m,3H),3.04-2.93(m,1H),2.51-2.45(m,1H),2.34(s,3H),2.33(s,3H),2.22(s,3H),2.11(s,3H),1.71-1.14(m,8H),0.83(t,J＝7.2Hz,3H).
实施例45：4-((4-((2,2-二氟乙基)(甲基)氨基)环己基)(乙基)氨基)-N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-1,5-二甲基-1H-吲唑-6-甲酰胺(P-45)的制备
Figure PCTCN2017110465-appb-000064
制备方法同化合物P-44，不同的是将步骤4中的化合物44.1换成2,2-二氟乙胺。经Pre-HPLC纯化得白色固体化合物P-45(123mg,71％),MS m/z(ESI):543.3[M+H]+。1H NMR(400MHz,DMSO)δ11.45(s,1H),8.15(t,J＝5.2Hz,1H),8.09(s,1H),7.35(s,1H),5.83(s,1H),4.31-4.26(m,4H),3.77-3.72(m,3H),3.59-3.56(m,2H),3.46-3.44(m,1H),3.28-3.12(m,4H),2.76-2.73(m,1H),2.24(s,3H),2.19(s,3H),2.07(s,3H),1.85-1.81(m,2H),1.63-1.57(m,2H),1.46-1.22(m,3H),0.74(t,J＝7.2Hz,3H).
实施例46：4-((4-氨基-4-甲基环己基)(乙基)氨基)-N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-1H-吲唑-6-甲酰胺(P-46)的制备
Figure PCTCN2017110465-appb-000065
步骤1至步骤4：制备方法同化合物P-22，不同的是将步骤1中化合物5a和2-二甲基环己酮换成化合物8a和24a。
步骤5：向化合物46-4(180mg,0.29mmol)的乙醇/盐酸(10ml/1ml,1M)的溶液中加入 钯炭(18mg),混合物在氢气氛围下室温搅拌过夜，LC-MS跟踪至反应结束。反应液过滤，滤液浓缩，经Pre-HPLC纯化得白色固体化合物P-46(19.11mg，14％),MS m/z(ESI):479.3[M+H]+。1H NMR(400MHz,DMSO)δ11.43(s,1H),8.40(s,1H),8.14(t,J＝4.0Hz,1H),8.07(s,1H),7.32(s,1H),5.87(s,1H),4.30(d,J＝4.0Hz,2H),3.98(s,3H),3.33-3.32(m,2H),3.05-3.04(m,1H),2.27(s,3H),2.23(s,3H),2.11(s,3H),1.59-1.58(m,4H),1.43-1.42(m,4H),1.19(s,3H),0.77(t,J＝8.0Hz,3H).
实施例47：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-5-乙基-4-(乙基(4-(3-羟基氮杂环丁烷-1-基)环己基)氨基)-1-甲基-1H-吲唑-6-甲酰胺(P-47)的制备
Figure PCTCN2017110465-appb-000066
步骤1至步骤4：制备方法同化合物P-22，不同的是将步骤1中化合物2-二甲基环己酮换成化合物22a。
步骤5：在-70℃下向化合物47-4(30mg,0.055mmol)的二氯甲烷溶液中慢慢滴加三溴化硼溶液(1M,0.27ml)，混合物在室温下搅拌2h，LC-MS跟踪至反应结束。向反应液中加入甲醇(10ml)，饱和碳酸氢钠溶液，用二氯甲烷萃取，有机相用无水硫酸钠干燥，过滤，浓缩，经柱层析纯化得米白色固体化合物P-47(7mg,24％),MS m/z(ESI):535.3[M+H]+。1H NMR(400MHz,DMSO)δ11.45(s,1H),8.12(t,J＝4.0Hz,1H),8.02(s,1H),7.27(s,1H),5.84(s,1H),5.20(br,1H),4.26(d,J＝4.0Hz,2H),4.04-4.03(m,1H),3.94(s,3H),3.39-3.38(m,2H),3.14-3.13(m,1H),2.89-2.87(m,3H),2.61-2.60(m,2H),2.20(s,3H),2.07(s,3H),1.91-1.81(m,2H),1.65-1.47(m,4H),1.30-1.20(m,2H),0.92(t,J＝8.0Hz,3H),0.75-0.72(m,5H).
实施例48：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(乙基(1-(氧杂环丁烷-3-基)哌啶-4-基)氨基)-1,5-二甲基-1H-吲唑-6-甲酰胺(P-48)的制备
Figure PCTCN2017110465-appb-000067
步骤1：制备方法同化合物22-1，不同的是将化合物5a和4-二甲基环己酮换成化合物8a和48.1。
步骤2：制备方法同化合物22-2，不同的是将化合物22-1换成48-1。
步骤3：向化合物48-2(358mg,0.77mmol)的甲醇溶液中加入催化量的盐酸(12M)和钯炭，混合物在氢气氛围下室温搅拌16h，LC-MS跟踪至反应结束。反应液过滤，滤液浓缩得化合物48-3(240mg),MS m/z(ESI):331[M+H]+。
步骤4：化合物48-3(240mg,0.72mmol),DIPEA(188mg,1.45mmol)的二氧六环溶液室温搅拌30min，加入化合物48.2(104mg,1.45mmol),混合物在室温下继续搅拌30min，加入醋酸硼氢化钠(231mg,1.09mmol),混合物在室温下继续搅拌3h，LC-MS跟踪至反应结束。用饱和碳酸氢钠溶液淬灭反应，并将PH调至8-9，反应液用二氯甲烷萃取，有机相用饱和食盐水洗涤，无水硫酸钠干燥，过滤，浓缩得化合物48-4(230mg),MS m/z(ESI):387[M+H]+。
步骤5：制备方法同化合物22-3，不同的是将化合物22-2换成化合物48-4。MS m/z(ESI):373[M+H]+。
步骤6：制备方法同化合物22-4，不同的是将化合物22-3和2a换成化合物48-4和1a。MS m/z(ESI):507[M+H]+。1H NMR(500MHz,DMSO)δ11.49(s,1H),8.15(t,J＝5.0Hz,1H),8.07(s,1H),7.31(s,1H),5.87(s,1H),4.47(t,J＝6.4Hz,2H),4.35(t,J＝6.0Hz,2H),4.30(d,J＝5.0Hz,2H),3.98(s,3H),3.28(m,2H),3.22–3.12(m,1H),3.09–3.02(m,1H),2.28(s,3H),2.23(s,3H),2.11(s,3H),1.88(m,1H),1.78–1.56(m,3H),1.48–1.31(m,2H),0.76(t,J＝7.1Hz,3H).
对比例1：N-((4,6-二甲基-2-氧代-1,2-二氢吡啶-3-基)甲基)-4-(乙基(四氢-2H-吡喃-4-基)氨基)-1,7-二甲基-1H-吲唑-6-甲酰胺(D1)的制备
Figure PCTCN2017110465-appb-000068
步骤1：制备方法同化合物22-1，不同的是将制法中的化合物5a和4-二甲氨基环己酮换成化合13a和四氢吡喃酮。MS m/z(ESI):304.2[M+H]+。
步骤2：制备方法同化合物22-2，不同的是将制法中的化合物22-1换成49-1。MS m/z(ESI):332.8[M+H]+。
步骤3：制备方法同化合物22-3，不同的是将制法中的化合物22-2换成49-2。MS m/z(ESI): 318.3[M+H]+。
步骤4：制备方法同化合物P-22，不同的是将制法中的化合物22-3和2a换成化合物49-3和1a。MS m/z(ESI):452[M+H]+。1H NMR(400MHz,DMSO)δ11.49(s,1H),8.05(t,J＝4.0Hz,1H),7.88(s,1H),6.36(s,1H),5.87(s,1H),4.28(d,J＝4.0Hz,2H),4.24(d,J＝4.0Hz,3H),3.88-3.85(m,2H),3.64-3.63(m,1H),3.29-3.23(m,4H),2.56(s,3H),2.21(s,3H),2.11(s,3H),1.68-1.66(m,4H),0.96(t,J＝8.0Hz,3H).
对比例2-4
对比化合物D2至D4参照对比化合物D1的方法制备
Figure PCTCN2017110465-appb-000069
Figure PCTCN2017110465-appb-000070
Figure PCTCN2017110465-appb-000071
Figure PCTCN2017110465-appb-000072
Figure PCTCN2017110465-appb-000073
Figure PCTCN2017110465-appb-000074
生物测试
测试例一体外甲基转移酶活性测试
重组PRC2(EZH2-Y641F)购自Active motif公司，S-甲硫腺苷氨酸(SAM)和L-多聚赖氨酸(PLL)购自Sigma-Aldrich公司，H3(1-50)K27me1多肽购自Cisbio公司。检测体系采用Perkinelmer公司的LANCEUltra系统。酶活实验中，将待测化合物按1:3的比例进行8个梯度点稀释后，加入反应板中并加100ng的重组酶。随后加入含有2.5μM SAM/250nM H3(1-50)K27me1预混物的缓冲液[20mM Tris pH8.5,2mM MgCl2,0.01％Tween-20,1mM TCEP]，室温下开始酶反应。反应3小时后，加入预混有PLL、检测抗体和Ulight的检测液，室温反应1小时后，在Tecan infinite pro上读取荧光值。IC50通过XLfit软件中的四因素模型拟合进行计算。结果如表1所示：
表1化合物对EZH2Y641F的抑制活性
Figure PCTCN2017110465-appb-000075
Figure PCTCN2017110465-appb-000076
测试例二细胞增殖测试
使用的细胞株Pfeiffer(CRL-2632)、suDHL-6(CRL-2959)、suDHL-10(CRL-2963)均购自美国菌种保藏库(ATCC)。所有细胞株以含10％胎牛血清(Gibco)的RPMI-1640培养基(Gbico)进行培养。培养细胞通过离心收集，并在CounterStar计数仪上测定细胞密度。随后在96孔板中种入合适数量的细胞并孵育过夜。待测化合物以1:3的比例进行8个梯度点稀释后，加入对应孔中。继续培养6天后，以Cell counting kit-8检测活细胞数，在Tecan infinite pro上读取吸光度值。IC50通过XLfit软件中的四参数模型拟合进行计算。结果如表2所示：
表2化合物对Pfeiffer细胞的抑制活性
Figure PCTCN2017110465-appb-000077
从表1和表2可以看出，本发明代表性化合物对EZH2的酶和细胞具有较高的抑制活性。研究发现，式(I)化合物中R0和R1的种类对化合物的酶抑制活性有很大影响，当R0为氢且R1为烷基时，对酶有很好的抑制活性，而当R0为烷基，R1为氢或其他取代基时，对酶的抑制活性大大降低。此外苯并五元杂环的种类对酶的抑制活性有较大影响，将化合物结构中式B-1换成式B-2、式B-3或式B-4时(例如P-3、P-4或P-50与P-7)，对酶的抑制活性明显降低。此外，化合物结构为式B-1时，其3位是否有取代基，以及其苯环5位取代基的变化，都对酶的抑制活性有较大影响，当3位的氢被甲基或氟取代后(例如P-42与P-13/14，P-41与P-7)，对酶的抑制活性大大降低，当苯环5位甲基换成其他取代基如氰基、环丙基时，对酶的抑制活性显著降低。
在本发明提及的所有文献都在本申请中引用作为参考，就如同每一篇文献被单独引用作为参考那样。此外应理解，在阅读了本发明的上述讲授内容之后，本领域技术人员可以对本发明作各种改动或修改，这些等价形式同样落于本申请所附权利要求书所限定的范围。

Claims (17)

  1. 一种式(I)所示的化合物，或其药学上可接受的盐、溶剂化物、立体异构体或前药：
    Figure PCTCN2017110465-appb-100001
    式中，
    R0为氢；
    R1为CN、卤素、C1-8烷基、卤代C1-8烷基、C1-8烷氧基、C3-8环烷基或C3-8环烷氧基；
    R2为氢、C1-8烷基、卤代C1-8烷基或C3-8环烷基；
    R3为氢、卤素、羟基、CN、C1-8烷基、卤代C1-8烷基、C3-8环烷基、C1-8烷氧基、卤代C1-8烷氧基、C3-8环烷氧基、C6-10芳基、-C(O)C1-8烷基、-C(O)OC1-8烷基、-CONRaRb或NRaRb；
    X为NR4、CR5R6、O、S或S(O)2；
    R4为氢、C1-8烷基、卤代C1-8烷基、C(O)C1-8烷基、C(O)OC1-8烷基、CONRa1Rb1、C3-8环烷基、-SO2C1-10烷基、-C(O)CH2CN、-C(O)CH2OH、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环；
    R5、R6各自独立地为氢、卤素、羟基、C1-8烷基、卤代C1-8烷基、C3-8环烷基、NRa2Rb2、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；
    Ra、Rb、Ra1、Rb1、Ra2、Rb2各自独立地为氢、C1-8烷基、C(O)C1-8烷基；
    A环为式(A-1)、式(A-2)、式(A-3)或式(A-4)所示结构：
    Figure PCTCN2017110465-appb-100002
    其中，R11、R21、R31、R41各自独立地为氢、C1-8烷基、卤代C1-8烷基、C3-8环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；
    R12、R22、R32、R42各自独立地为氢、卤素、C1-8烷基、卤代C1-8烷基或C3-8环烷基；
    所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1、2或3个选自下组的取代基所取代：CN、乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OC1-6烷基、C1-8烷基、C1-8烷氧基、卤代C1-8烷基、C3-8环烷基、卤代C1-8烷氧基、NRa3Rb3、4至6元饱和单杂环、5至6 元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环；其中Ra3、Rb3各自独立地为氢、C1-8烷基或C1-8烷氧基取代的C1-8烷基。
  2. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R1为C1-3烷基。
  3. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R2为氢、C1-3烷基或卤代C1-3烷基。
  4. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R3为氢、卤素、C1-3烷基或C1-3烷氧基。
  5. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R0为氢且R1为C1-3烷基。
  6. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R11、R21、R31、R41各自独立地为氢或-(CH2)n-L1；其中L1为CN、NRa3Rb3、C1-8烷基、C1-8烷氧基、卤代C1-8烷基、C3-8环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；n为0、1或2；
    所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OC1-6烷基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、NRa3Rb3、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃；Ra3、Rb3如权利要求1所定义。
  7. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R12、R22、R32、R42各自独立地为氢、甲基、乙基、正丙基、异丙基。
  8. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R4为氢或-(CH2)m-L2；其中L2为CN、C1-8烷基、NRa3Rb3、C1-8烷氧基、卤代C1-8烷基、C(O)C1-8烷基、C(O)OC1-8烷基、CONRa1Rb1、C3-8环烷基、-SO2C1-8烷基、-C(O)CH2CN、-C(O)CH2OH、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环；m为0、1或2；
    所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、8至10元双环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1个选自下组的取代基所取代：乙酰基、羟基、羟甲基、羟乙基、羧基、-C(O)OC1-6烷基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、NRa3Rb3、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃；Ra3、Rb3如权利要求1所定义。
  9. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，R5为氢且R6为-(CH2)p-L3；L3为CN、C1-8烷基、C1-8烷氧基、卤代C1-8烷基、C3-8环烷基、NRa3Rb3、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环；p为0、1或2；
    所述烷基、烷氧基、环烷基、4至6元饱和单杂环、5至6元单环杂芳基环、螺环、螺杂环、桥环或桥杂环为未取代的或被1个选自下组的取代基所取代：乙酰基、羟基、羟甲基、 羟乙基、羧基、-C(O)OC1-6烷基、C1-3烷基、C1-3烷氧基、卤代C1-3烷基、C3-6环烷基、NRa3Rb3、氮杂环丁烷、氧杂环丁烷、四氢呋喃、四氢噻吩、四氢吡咯、哌啶、噁唑烷、哌嗪、二氧戊环、二氧六环、吗啉、硫代吗啉、硫代吗啉-1,1-二氧化物或四氢吡喃；Ra3、Rb3如权利要求1所定义。
  10. 如权利要求1所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，其特征在于，
    Figure PCTCN2017110465-appb-100003
    为式(B-1)、式(B-2)、式(B-3)或式(B-4)所示结构：
    Figure PCTCN2017110465-appb-100004
    其中R11、R12、R21、R22、R31、R32、R41、R42如权利要求1所定义。
  11. 如权利要求1所述的化合物、或其药学上可接受的盐、或其溶剂化物、或其立体异构体、或其前药，其特征在于，所述化合物选自表A。
  12. 一种药物组合物，所述药物组合物包括权利要求1至11中任一项所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药；以及药学可接受的载体。
  13. 如权利要求1至11中任一项所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药、或如权利要求12所述药物组合物在制备EZH2抑制剂的应用。
  14. 如权利要求1至11中任一项所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药、或如权利要求12所述药物组合物在制备由EZH2介导的疾病或病症的应用。
  15. 一种治疗由EZH2介导的疾病或病症的方法，包括给予所需患者治疗有效量的权利要求1至11中任一项所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，或如权利要求12所述药物组合物。
  16. 一种治疗由EZH2介导的疾病或病症的方法，包括给予所需患者治疗有效量的权利要求1至11中任一项所述的化合物、或其药学上可接受的盐、溶剂化物、立体异构体或前药，以及另一种治疗活性试剂。
  17. 如权利要求1所述的化合物、或其药学上可接受的盐、或其溶剂化物、或其立体异构体、或其前药，其特征在于，所述化合物选自表B。
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