WO2024153155A1 - 双杂环wrn抑制剂、其制备方法及其应用 - Google Patents

双杂环wrn抑制剂、其制备方法及其应用 Download PDF

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WO2024153155A1
WO2024153155A1 PCT/CN2024/072876 CN2024072876W WO2024153155A1 WO 2024153155 A1 WO2024153155 A1 WO 2024153155A1 CN 2024072876 W CN2024072876 W CN 2024072876W WO 2024153155 A1 WO2024153155 A1 WO 2024153155A1
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membered
independently
alkyl
substituted
membered heteroaryl
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PCT/CN2024/072876
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English (en)
French (fr)
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王伟
孙晓阳
刘自强
张继鑫
卿盈
张萍
刘航
山松
潘德思
鲁先平
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成都微芯药业有限公司
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Publication of WO2024153155A1 publication Critical patent/WO2024153155A1/zh

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  • the present invention belongs to the field of pharmaceutical chemistry, and in particular relates to a biheterocyclic WRN inhibitor, a preparation method thereof and an application thereof.
  • WRN is a member of the RecQ family of DNA helicases and is involved in DNA damage repair, genome stability maintenance, telomere maintenance, etc. WRN gene defects can lead to Werner syndrome (progeria syndrome), a recessive genetic disease characterized by staged premature aging and increased cancer susceptibility (Nat Rev Cancer 2003, 3, 169-178). In 2019, Cancer Dependency Map (DepMap), a project aimed at finding new tumor treatment targets through gene screening, drug sensitivity and drug prediction models, found that WRN is essential for the survival of microsatellite unstable (MSI) tumors.
  • MSI microsatellite unstable
  • WRN can unwind this structure to restore normal DNA replication.
  • the cross-shaped structure will be cut by the nuclease MUS81, resulting in chromosome genome damage and cell death.
  • the TA repeat sequence does not form a cross-shaped structure, so WRN deficiency will specifically cause MSI cell death (Nature 2020, 586, 292-298). Therefore, inhibitors targeting the WRN unwinding domain may be an effective measure to treat MSI cancers.
  • WRN inhibitors Although some patent applications for WRN inhibitors have been published, new compounds still need to be further developed, as there are currently no WRN target drugs on the market and WRN target inhibition has potential medical value in the treatment of microsatellite instability (MSI) tumors.
  • MSI microsatellite instability
  • the inventors of the present application designed a compound having a structure shown in general formula (I), and found that compounds having such a structure exhibit excellent WRN inhibition effects and functions, and can be used to treat MSI tumor-related diseases.
  • R 1 is selected from halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 5-membered or 6-membered cycloalkenyl, 5-10-membered heterocyclyl, 5-membered or 6-membered heteroaryl, phenyl, wherein the heterocyclyl and heteroaryl each independently contain 1-2 heteroatoms selected from N, O or S, and the alkyl, alkenyl, alkynyl, cycloalkenyl, heterocyclyl, heteroaryl, phenyl each independently is unsubstituted or substituted by 1 or more identical or different R g ;
  • R 2 is selected from Wherein, L is selected from 7-10 membered spiroheterocycloalkyl, The spiroheterocycloalkyl group contains 2-3 N atoms;
  • R 3 is selected from cyclopropyl, methoxy, -SCH 3 , C 1-4 alkyl, said alkyl being unsubstituted or substituted by one or more substituents selected from hydroxyl or halogen;
  • R 4 is selected from H, halogen, hydroxyl, cyano, C 1-4 alkyl, C 3-5 cycloalkyl, C 1-4 alkoxy, wherein the alkyl, cycloalkyl, alkoxy is unsubstituted or substituted by one or more halogens;
  • R 5 is selected from H, methoxy, hydroxy, cyano, methyl, halogen
  • R6 is selected from H, halogen, C1-4 alkyl; the alkyl is unsubstituted or substituted by one or more halogens;
  • R7 is selected from H, -SF5 , -C(O)H, halogen, C1-4 alkyl, C2-4 alkenyl, C2-4 alkynyl, C3-5 cycloalkyl, C1-4 alkoxy; the alkyl, cycloalkyl, alkoxy is unsubstituted or substituted by one or more halogens. generation;
  • X is selected from N or CR 8 , R 8 is H or halogen;
  • R 2a is selected from 6-14 membered aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the aryl, heteroaryl and heterocyclic group are unsubstituted or each independently substituted by one or more identical or different R c ;
  • R 2e is selected from 5-14 membered heteroaryl, 6-14 membered aryl, 5-14 membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the heteroaryl, aryl and heterocyclic group are unsubstituted or each independently substituted by one or more identical or different R f ;
  • Each R c is independently selected from halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 3-6 halocycloalkyl, cyano, amino;
  • Each Rd is independently selected from C1-6 alkyl, C3-6 cycloalkyl, C1-6 haloalkyl, cyano, amino, C1-6 alkoxy, C1-6 haloalkoxy, C3-6 halocycloalkyl, hydroxy, halogen, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl; the 4-7 membered heterocycloalkyl contains 1 or 2 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl each independently contains 1 to 3 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl, phenyl is unsubstituted or is independently substituted with one or more substituents selected from halogen, cyano, C1-3 alkyl, C1-3 haloalkyl, C1-3 alkoxy, C1-3 haloalkoxy, C3-5 cycloalkyl;
  • Each R f is independently selected from hydroxy, cyano, amino, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy , C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 3-6 halocycloalkyl.
  • R 1 is selected from halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 5-membered or 6-membered cycloalkenyl, 5-membered or 6-membered heterocyclyl, 5-membered or 6-membered heteroaryl, phenyl, wherein the heterocyclyl or heteroaryl contains 1-2 heteroatoms selected from N, O or S, and the alkyl, alkenyl, alkynyl, cycloalkenyl, heterocyclyl, heteroaryl, phenyl is unsubstituted or substituted by 1 or more identical or different R g ;
  • R 2 is selected from Wherein, L is selected from 7-10 membered spiro heterocyclic group,
  • R 3 is selected from cyclopropyl, methoxy, -SCH 3 , C 1-4 alkyl, said alkyl being unsubstituted or substituted by one or more substituents selected from hydroxyl or halogen;
  • R 4 is selected from H, halogen, hydroxyl, cyano, C 1-4 alkyl, C 3-5 cycloalkyl, C 1-4 alkoxy, wherein the alkyl, cycloalkyl, alkoxy is unsubstituted or substituted by one or more halogens;
  • R 5 is selected from H, methoxy, hydroxy, cyano, methyl, halogen
  • R6 is selected from H, halogen, C1-4 alkyl; the alkyl is unsubstituted or substituted by one or more halogens;
  • R 7 is selected from H, -SF 5 , -C(O)H, halogen, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-5 cycloalkyl, C 1-4 alkoxy; the alkyl, cycloalkyl, alkoxy is unsubstituted or substituted by one or more halogens;
  • X is selected from N or CR 8 , R 8 is H or halogen;
  • R 2a is selected from 6-14 membered aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the aryl, heteroaryl and heterocyclic group are unsubstituted or each independently substituted by one or more identical or different R c ;
  • R 2b is selected from 5-membered heteroaryl, 8-14-membered heteroaryl, 6-14-membered aryl, 5-14-membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the 5-membered heteroaryl, 8-14-membered heteroaryl, 6-14-membered aryl, 5-14-membered heterocyclic group is each independently substituted by one or more R d ;
  • R 2e is selected from 5-14 membered heteroaryl, 6-14 membered aryl, 5-14 membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the heteroaryl, aryl and heterocyclic group are unsubstituted or each independently substituted by one or more identical or different R f ;
  • Each R c is independently selected from halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 3-6 halocycloalkyl, cyano, amino;
  • Each Rd is independently selected from C1-6 alkyl, C3-6 cycloalkyl, C1-6 haloalkyl, cyano, amino, C1-6 alkoxy, C1-6 haloalkoxy, C3-6 halocycloalkyl, 4-7 membered heterocycloalkyl, hydroxyl or halogen; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S;
  • Each R f is independently selected from hydroxy, cyano, amino, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy , C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 3-6 halocycloalkyl.
  • R 1 is selected from halogen, C 1-6 alkyl, C 2-6 alkenyl, C 2-6 alkynyl, 5-membered or 6-membered cycloalkenyl, 5-membered or 6-membered heterocyclyl, 5-membered or 6-membered heteroaryl, phenyl, wherein the heterocyclyl or heteroaryl contains 1-2 heteroatoms selected from N, O or S, and the alkyl, alkenyl, alkynyl, cycloalkenyl, heterocyclyl, heteroaryl, phenyl is unsubstituted or substituted by 1 or more identical or different R g ;
  • R 2 is selected from Wherein, L is selected from 7-10 membered spiro heterocyclic group,
  • R 3 is selected from cyclopropyl, methoxy, -SCH 3 , C 1-4 alkyl, said alkyl being unsubstituted or substituted by one or more substituents selected from hydroxyl or halogen;
  • R 4 is selected from H, halogen, hydroxyl, cyano, C 1-4 alkyl, C 3-5 cycloalkyl, C 1-4 alkoxy, wherein the alkyl, cycloalkyl, alkoxy is unsubstituted or substituted by one or more halogens;
  • R 5 is selected from H, methoxy, hydroxy, cyano, methyl, halogen
  • R6 is selected from H, halogen, C1-4 alkyl; the alkyl is unsubstituted or substituted by one or more halogens;
  • R 7 is selected from H, -SF 5 , -C(O)H, halogen, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-5 cycloalkyl, C 1-4 alkoxy; the alkyl, cycloalkyl, alkoxy is unsubstituted or substituted by one or more halogens;
  • X is selected from N or CR 8 , R 8 is H or halogen;
  • R 2a is selected from 6-14 membered aryl, 5-14 membered heteroaryl, 5-14 membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the aryl, heteroaryl and heterocyclic group are unsubstituted or each independently substituted by one or more identical or different R c ;
  • R 2b is selected from 5-membered heteroaryl, 8-14-membered heteroaryl, 6-14-membered aryl, 5-14-membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the 5-membered heteroaryl, 8-14-membered heteroaryl, 6-14-membered aryl, 5-14-membered heterocyclic group is each independently substituted by one or more R d ;
  • R 2e is selected from 5-14 membered heteroaryl, 6-14 membered aryl, 5-14 membered heterocyclic group, wherein the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the heteroaryl, aryl and heterocyclic group are unsubstituted or each independently substituted by one or more identical or different R f ;
  • Each R c is independently selected from halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 3-6 halocycloalkyl, cyano, amino;
  • Each R d is independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, cyano, amino, C 1-6 alkoxy, C 1-6 haloalkoxy, C 3-6 halocycloalkyl, hydroxyl or halogen;
  • Each R f is independently selected from hydroxy, cyano, amino, halogen, C 1-6 alkyl, C 1-6 haloalkyl, C 1-6 alkoxy , C 1-6 haloalkoxy, C 3-6 cycloalkyl, C 3-6 halocycloalkyl.
  • R 2 is selected from
  • R2 is selected from
  • R2 is selected from
  • R2 is selected from
  • R2 is selected from
  • R2 is selected from
  • R2 is selected from
  • R2 is selected from
  • R 2b is selected from 5-membered heteroaryl and 8-14-membered heteroaryl; the 5-membered heteroaryl and 8-14-membered heteroaryl each independently contain 1-4 heteroatoms selected from N, O and S; the 5-membered heteroaryl and 8-14-membered heteroaryl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, and 10-membered heteroaryl; the 5-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, and 10-membered heteroaryl each independently contain 1-4 heteroatoms selected from N, O and S; the 5-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, and 10-membered heteroaryl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, 5-membered heteroaryl and 5-membered heteroaryl, benzo 5-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, benzo 6-membered heteroaryl, 6-membered heteroaryl and 6-membered heteroaryl; the 5-membered heteroaryl and 6-membered heteroaryl each independently contain 1-2 heteroatoms selected from N, O and S; the 5-membered heteroaryl, 5-membered heteroaryl and 5-membered heteroaryl, benzo 5-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, benzo 6-membered heteroaryl, 6-membered heteroaryl and 6-membered heteroaryl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, benzo 5-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, benzo 6-membered heteroaryl; the 5-membered heteroaryl and 6-membered heteroaryl each independently contain 1-2 heteroatoms selected from N, O and S; the 5-membered heteroaryl, benzo 5-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, benzo 6-membered heteroaryl are each independently substituted by one or more R d .
  • R 2b is selected from pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, benzopyrrolyl, benzofuranyl, benzothienyl, benzopyrazolyl, benzimidazolyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl, pyridopyrrolyl, pyridofuranyl, pyridothiphenyl, pyridopyrazolyl, pyridoimidazolyl, pyridothi ...
  • pyridothiazolyl pyridoisothiazolyl, pyridooxazolyl, pyridoisoxazolyl, benzopyridinyl, benzopyrimidinyl; the pyrrolyl, furanyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, benzopyrrolyl, benzofuranyl, benzothienyl, benzopyrazolyl, benzimidazolyl, benzothiazolyl, benzisothiazolyl, benzoxazolyl, benzisoxazolyl.
  • R 2b is selected from pyrazolyl, benzopyridinyl, isoxazolyl, isothiazolyl, pyridopyrazolyl, pyridoimidazolyl, pyridofuranyl, pyridopyrrolyl, pyridothiphenyl; and the pyrazolyl, benzopyridinyl, isoxazolyl, isothiazolyl, pyridopyrazolyl, pyridoimidazolyl, pyridofuranyl, pyridopyrrolyl, pyridothiphenyl is each independently substituted by one or more R d .
  • R 2b is selected from Said are each independently substituted with one or more R d .
  • R 2b is selected from
  • each R d is independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, hydroxyl, halogen, wherein one and only one R d is hydroxyl; the 4-7 membered heterocycloalkyl contains 1 or 2 heteroatoms selected from N, O and S; the 5-membered or 6-membered heteroaryl each independently contains 1 to 3 heteroatoms selected from N, O and S.
  • each R d is independently selected from C 1-4 alkyl, C 5-6 alkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, C 1-4 haloalkyl, C 5-6 haloalkyl, C 1-4 alkoxy, C 5-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, hydroxyl, halogen, wherein there is only one R d is hydroxy; the 4-7 membered heterocycloalkyl contains 1 or 2 heteroatoms selected from N, O and S; the 5-membered or 6-membered heteroaryl each independently contains 1, 2 or 3 heteroatoms selected from N, O and S.
  • each R d is independently selected from C 1-4 alkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, C 1-4 haloalkyl, C 1-4 alkoxy, 4-7 membered heterocycloalkyl, 6-membered heteroaryl, phenyl, hydroxyl, halogen, wherein one and only one R d is hydroxyl; the 4-7 membered heterocycloalkyl contains 1 or 2 O heteroatoms; the 6-membered heteroaryl contains 1 or 2 N heteroatoms.
  • each R d is independently selected from C 1-2 alkyl, C 3-4 alkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl , C 1-2 fluoroalkyl, C 3-4 fluoroalkyl, C 1-2 alkoxy, C 3-4 alkoxy, 4-7 membered heterocycloalkyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, phenyl, hydroxyl, halogen, wherein one and only one R d is hydroxyl; and the 4-7 membered heterocycloalkyl contains 1 O heteroatom.
  • each R d is independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, fluoromethyl, fluoroethyl, fluoropropyl, fluoroisopropyl, methoxy, ethoxy, 4-7 membered heterocycloalkyl, pyridyl, hydroxyl, F, Cl, Br, phenyl, wherein one and only one R d is hydroxyl; and the 4-7 membered heterocycloalkyl contains 1 O heteroatom.
  • each R d is independently selected from methyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, -CH 2 CF 3 , Methoxy, Cl, phenyl, hydroxy, wherein one and only one R d is hydroxy.
  • the hydroxyl group is substituted on the carbon atom at the ortho or meta position of the connection site between R 2b and the main group.
  • the hydroxyl group is substituted on the carbon atom adjacent to the connection site of R 2b and the main group.
  • R 2b is selected from
  • each R d is independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, halogen;
  • the 4-7 membered heterocycloalkyl group contains 1 or 2 heteroatoms selected from N, O and S; the 5-membered or 6-membered heteroaryl group each independently contains 1 to 3 heteroatoms selected from N, O and S.
  • each Rd is independently selected from C1-4 alkyl, C5-6 alkyl, C3-4 cycloalkyl, C5-6 cycloalkyl, C1-4 haloalkyl, C5-6 haloalkyl, C1-4 alkoxy , C5-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, halogen; the 4-7 membered heterocycloalkyl contains 1 or 2 heteroatoms selected from N, O and S; the 5-membered or 6-membered heteroaryl each independently contains 1, 2 or 3 heteroatoms selected from N, O and S.
  • each Rd is independently selected from C1-4 alkyl, C3-4 cycloalkyl, C5-6 cycloalkyl, C1-4 haloalkyl, C1-4 alkoxy, 4-7 membered heterocycloalkyl, 6 membered heteroaryl, phenyl, halogen; the 4-7 membered heterocycloalkyl contains 1 or 2 O heteroatoms; the 6 membered heteroaryl contains 1 or 2 N heteroatoms.
  • each Rd is independently selected from C1-2 alkyl, C3-4 alkyl, C3-4 cycloalkyl, C5-6 cycloalkyl, C1-2 fluoroalkyl, C3-4 fluoroalkyl, C1-2 alkoxy , C3-4 alkoxy, 4-7 membered heterocycloalkyl, pyridyl, pyrimidinyl, pyridazinyl, pyrazinyl, phenyl, halogen; the 4-7 membered heterocycloalkyl contains 1 O heteroatom.
  • each Rd is independently selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, fluoromethyl, fluoroethyl, fluoropropyl, fluoroisopropyl, methoxy, ethoxy, 4-7 membered heterocycloalkyl, pyridyl, F, Cl, Br, phenyl; the 4-7 membered heterocycloalkyl contains 1 O heteroatom.
  • each R d is independently selected from methyl, isopropyl, tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, -CH 2 CF 3 , Methoxy, Cl, phenyl.
  • R 2b is selected from
  • R 2b is selected from 5-membered heteroaryl, phenyl and 5- or 6-membered heteroaryl, 5- or 6-membered heteroaryl and 5- or 6-membered heteroaryl, 5- or 6-membered heterocycloalkyl, 5- or 6-membered heterocycloalkenyl, 5- or 6-membered heterocycloalkyl and 5- or 6-membered heterocycloalkenyl; the -CH 2 - on the heterocycloalkyl and heterocycloalkenyl rings are each independently optionally replaced by -C( ⁇ O)-; the heteroaryl, heterocycloalkyl and heterocycloalkenyl each independently contain 1-2 heteroatoms selected from N, O or S; the 5-membered heteroaryl, phenyl and 5- or 6-membered heteroaryl, 5- or 6-membered heteroaryl and 5- or 6-membered heteroaryl, 5- or 6-membered heterocycloalkyl, 5- or 6-membered heterocycloalkenyl,
  • R 2b is selected from 5-membered heteroaryl, phenyl and 5-membered or 6-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, 1,5-dihydro-2H-pyrrole-2-one, pyridone, 6-membered heterocycloalkyl and Pyridonyl, the heteroaryl and heterocycloalkyl each independently contain 1-2 heteroatoms selected from N, O or S; the 5-membered heteroaryl, phenyl and 5-membered or 6-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, 1,5-dihydro-2H-pyrrole-2-one, pyridonyl, 6-membered heterocycloalkyl and pyridonyl are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from isoxazolyl, pyrazolyl, isothiazolyl, benzopyrrolyl, benzopyridinyl, morpholinopyridinonyl, 1,5-dihydro-2H-pyrrol-2-onyl,
  • the isoxazolyl, pyrazolyl, isothiazolyl, benzopyrrolyl, benzopyridyl, morpholinopyridone, 1,5-dihydro-2H-pyrrol-2-one, are each independently substituted with 1, 2, 3 or 4 R d .
  • R 2b is selected from isoxazolyl, pyrazolyl, isothiazolyl, The isoxazolyl, pyrazolyl, isothiazolyl, are each independently substituted with 1, 2, 3 or 4 R d .
  • R 2b is selected from Said are each independently substituted with 1, 2, 3 or 4 R d .
  • each Rd is independently selected from halogen, C1-6 alkyl, C3-6 cycloalkyl, C1-6 haloalkyl, cyano, C1-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, amino or hydroxyl; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl each independently contains 1 to 3 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl, phenyl is unsubstituted or is independently substituted with one or more substituents selected from halogen, cyano, C1-3 alkyl, C1-3 haloalkyl, C1-3 alkoxy, C1-3 haloalkoxy, C3-5 cycloalkyl.
  • each Rd is independently selected from halogen, C1-4 alkyl, C5-6 alkyl, C3-4 cycloalkyl, C5-6 cycloalkyl, C1-4 haloalkyl, C5-6 haloalkyl, C1-4 alkoxy , C5-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, cyano or hydroxyl;
  • the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S;
  • the 5-membered or 6-membered heteroaryl each independently contains 1, 2 or 3 heteroatoms selected from N, O or S;
  • the 5-membered or 6-membered heteroaryl, phenyl is unsubstituted or is independently substituted with one or more substituents selected from halogen, cyano, C1-3 alkyl, C1-3 haloalkyl, C1-3 alkoxy, C1-3 halo
  • each Rd is independently selected from halogen, C1-4 alkyl, C3-4 cycloalkyl, C5-6 cycloalkyl, C1-4 haloalkyl, C1-4 alkoxy, 4-7 membered heterocycloalkyl, phenyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, furanyl, oxazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, cyano or hydroxy; the 4-7 membered heterocycloalkyl each independently contains 1 O heteroatom; the phenyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, furanyl, oxazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl is unsubstituted or substituted with one or more substituents selected
  • each R d is independently selected from phenyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, furanyl, oxazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, methyl, ethyl, propyl, isopropyl, tert-butyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, -CH 2 CF 3 , Methoxy, ethoxy, Fluorine-substituted phenyl, chlorine-substituted phenyl, cyano-substituted phenyl, methyl-substituted phenyl, methoxy-substituted phenyl, trifluoromethyl-substituted phenyl, trifluoromethoxy-substituted phenyl,
  • each R d is independently selected from phenyl, thienyl, pyridyl, hydroxyl, methyl, isopropyl, tert-butyl, cyclobutyl, cyclopentyl, cyclopropyl, -CH 2 CF 3 , Methoxy, or Cl.
  • each R d is independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, C1-6 haloalkyl, cyano, amino, C1-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, hydroxyl or halogen, wherein at least one Rd is hydroxyl; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl each independently contains 1 to 3 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl, phenyl is unsubstituted or is each independently substituted by one or more substituents selected from halogen, cyano, C1-3 alkyl, C1-3 haloalkyl, C1-3 alkoxy, C1-3 haloalkoxy, C3-5 cycloalkyl.
  • each Rd is independently selected from halogen, C1-6 alkyl, C3-6 cycloalkyl, C1-6 haloalkyl, C1-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, cyano or hydroxy, wherein at least one Rd is hydroxy;
  • the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S;
  • the 5-membered or 6-membered heteroaryl each independently contains 1 to 3 heteroatoms selected from N, O or S;
  • the 5-membered or 6-membered heteroaryl, phenyl is unsubstituted or is independently substituted with one or more substituents selected from halogen, cyano, C1-3 alkyl, C1-3 haloalkyl, C1-3 alkoxy, C1-3 haloalkoxy, C3-5 cycloalkyl.
  • each R d is independently selected from halogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl , C 1-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, cyano or hydroxy, wherein one and only one R d is hydroxy;
  • the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S;
  • the 5-membered or 6-membered heteroaryl each independently contains 1 to 3 heteroatoms selected from N, O or S;
  • the 5-membered or 6-membered heteroaryl, phenyl is unsubstituted or is independently substituted with one or more substituents selected from halogen, cyano, C 1-3 alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 haloalkoxy, C 3-5 cycloalkyl.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 5-6 alkyl, C 1-4 haloalkyl, C 5-6 haloalkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, C 1-4 alkoxy, C 5-6 alkoxy, 4-7 membered heterocycloalkyl, 5-membered or 6-membered heteroaryl, phenyl, cyano or hydroxy, wherein one and only one R d is hydroxy; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl each independently contains 1, 2 or 3 heteroatoms selected from N, O or S; the 5-membered or 6-membered heteroaryl, phenyl is unsubstituted or is independently substituted with one or more substituents selected from halogen, cyano, C 1-3 alkyl, C 1-3 hal
  • each R d is independently selected from halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, C 1-4 alkoxy, 4-7 membered heterocycloalkyl, phenyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, furanyl, oxazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, cyano or hydroxy, wherein one and only one R d is hydroxy; the 4-7 membered heterocycloalkyl each independently contains 1 O heteroatom; the phenyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, furanyl, oxazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl is
  • each R d is independently selected from phenyl, thienyl, thiazolyl, pyrazolyl, pyrrolyl, furanyl, oxazolyl, pyridinyl, pyrimidinyl, pyridazinyl, pyrazinyl, methyl, ethyl, propyl, isopropyl, tert-butyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, -CH 2 CF 3 , Methoxy, ethoxy, Fluorine-substituted phenyl, chlorine-substituted phenyl, cyano-substituted phenyl, methyl-substituted phenyl, methoxy-substituted phenyl, trifluoromethyl-substituted phenyl, trifluoromethoxy-substituted phenyl,
  • each R d is independently selected from phenyl, thienyl, pyridyl, hydroxyl, methyl, isopropyl, tert-butyl, cyclobutyl, cyclopentyl, cyclopropyl, -CH 2 CF 3 , Methoxy, or Cl, wherein one and only one R d is a hydroxyl group.
  • each R d is independently selected from phenyl, thienyl, pyridyl, hydroxyl, methyl, isopropyl, tert-butyl, cyclobutyl, cyclopentyl, cyclopropyl, -CH 2 CF 3 , Methoxy, Or Cl, wherein one and only one R d is a hydroxyl group, and the hydroxyl group is substituted at the ortho position of the site where R 2b is connected to the main group.
  • R 2b is selected from
  • R 2b is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2 is selected from
  • R 2b is selected from 5-membered heteroaryl, 8-10-membered heteroaryl, 6-10-membered aryl, 5-10-membered heterocyclic group, and the heteroaryl and heterocyclic group each independently contain 1-4 heteroatoms selected from N, O or S; the 5-membered heteroaryl, 8-10-membered heteroaryl, 6-10-membered aryl, 5-10-membered heterocyclic group is each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, 8-10-membered heteroaryl, phenyl, naphthyl, 5-membered or 6-membered heterocyclyl, 8-10-membered heterocyclyl, and the heteroaryl and heterocyclyl each independently contain 1-4 heteroatoms selected from N, O or S; the 5-membered heteroaryl, 8-10-membered heteroaryl, phenyl, naphthyl, 5-membered or 6-membered heterocyclyl, 8-10-membered heterocyclyl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, 8-10-membered heteroaryl, 5-membered or 6-membered heterocyclyl, 8-10-membered heterocyclyl, and the heteroaryl and heterocyclyl each independently contain 1-4 heteroatoms selected from N, O or S; and the 5-membered heteroaryl, 8-10-membered heteroaryl, 5-membered or 6-membered heterocyclyl, and 8-10-membered heterocyclyl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, phenyl and 5- or 6-membered heteroaryl, 5- or 6-membered heteroaryl and 5- or 6-membered heteroaryl, 5- or 6-membered heterocyclyl, 5- or 6-membered heterocyclyl and 5- or 6-membered heterocyclyl; the heteroaryl and heterocyclyl each independently contain 1-2 heteroatoms selected from N, O or S; the 5-membered heteroaryl, phenyl and 5- or 6-membered heteroaryl, 5- or 6-membered heteroaryl and 5- or 6-membered heteroaryl, 5- or 6-membered heterocyclyl, 5- or 6-membered heterocyclyl and 5- or 6-membered heterocyclyl are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from 5-membered heteroaryl, phenyl and 5-membered or 6-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, 5-membered heterocyclyl, 6-membered heterocyclyl, 6-membered heterocyclyl and pyridinone, and the heteroaryl and heterocyclyl each independently contain 1-2 heteroatoms selected from N, O or S; and the 5-membered heteroaryl, phenyl and 6-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, 5-membered heterocyclyl, 6-membered heterocyclyl, 6-membered heterocyclyl and pyridinone are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from isoxazolyl, pyrazolyl, isothiazolyl, benzopyrrolyl, benzopyridinyl, morpholinopyridinonyl, 1,5-dihydro-2H-pyrrol-2-onyl,
  • the isoxazolyl, pyrazolyl, isothiazolyl, benzopyrrolyl, benzopyridyl, morpholinopyridone, 1,5-dihydro-2H-pyrrol-2-one are each independently substituted with 1, 2, 3 or 4 R d .
  • each R d is independently selected from halogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, cyano, C 1-6 alkoxy, 4-7 membered heterocycloalkyl, amino or hydroxy; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S.
  • each Rd is independently selected from halogen, C1-4 alkyl, C5-6 alkyl, C3-4 cycloalkyl, C5-6 cycloalkyl, C1-4 haloalkyl, C5-6 haloalkyl, C1-4 alkoxy, C5-6 alkoxy, 4-7 membered heterocycloalkyl, cyano or hydroxy; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-4 haloalkyl, C 1-4 alkoxy, 4-7 membered heterocycloalkyl, cyano or hydroxy; and the 4-7 membered heterocycloalkyl each independently contains 1 O heteroatom.
  • each R d is independently selected from methyl, ethyl, propyl, isopropyl, tert-butyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, -CH 2 CF 3 , Methoxy, ethoxy, or hydroxyl.
  • each R d is independently selected from hydroxy, methyl, isopropyl, tert-butyl, cyclobutyl, cyclopentyl, cyclopropyl, -CH 2 CF 3 , Methoxy, or Cl.
  • each Rd is independently selected from C1-6 alkyl, C3-6 cycloalkyl, C1-6 haloalkyl, cyano, amino, C1-6 alkoxy , 4-7 membered heterocycloalkyl, hydroxyl or halogen, wherein at least one Rd is hydroxyl; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S.
  • each R d is independently selected from halogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, 4-7 membered heterocycloalkyl, cyano or hydroxy, wherein at least one R d is hydroxy; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S.
  • each R d is independently selected from halogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, C 1-6 alkoxy, 4-7 membered heterocycloalkyl, cyano or hydroxy, wherein one and only one R d is hydroxy; the 4-7 membered heterocycloalkyl each independently contains 1 or 2 heteroatoms selected from N, O or S.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 5-6 alkyl, C 1-4 haloalkyl, C 5-6 haloalkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, C 1-4 alkoxy, C 5-6 alkoxy, 4-7 membered heterocycloalkyl, cyano or hydroxyl, wherein one and only one R d is hydroxyl; the 4-7 membered hetero
  • the cycloalkyl groups each independently contain 1 or 2 heteroatoms selected from N, O or S.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl, C 1-4 alkoxy, 4-7 membered heterocycloalkyl, cyano or hydroxy, wherein one and only one R d is hydroxy; and the 4-7 membered heterocycloalkyl each independently contains 1 O heteroatom.
  • each R d is independently selected from methyl, ethyl, propyl, isopropyl, tert-butyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, -CH 2 CF 3 , Methoxy, ethoxy, or hydroxyl, wherein one and only one R d is hydroxyl.
  • each R d is independently selected from hydroxy, methyl, isopropyl, tert-butyl, cyclobutyl, cyclopentyl, cyclopropyl, -CH 2 CF 3 , Methoxy, or Cl, wherein one and only one R d is a hydroxyl group.
  • each R d is independently selected from hydroxy, methyl, isopropyl, tert-butyl, cyclobutyl, cyclopentyl, cyclopropyl, -CH 2 CF 3 , Methoxy, Or Cl, wherein one and only one R d is a hydroxyl group, and the hydroxyl group is substituted at the ortho position of the site where R 2b is connected to the main group.
  • R 2b is selected from
  • R2 is selected from
  • R2 is selected from
  • R2 is selected from
  • R 2b is selected from 5-membered heteroaryl, 8-10-membered heteroaryl, 6-10-membered aryl, 5-10-membered heterocyclyl, and the heteroaryl and heterocyclyl each independently contain 1-4 heteroatoms selected from N, O or S; and the 5-membered heteroaryl, 8-10-membered heteroaryl, 6-10-membered aryl, 5-10-membered heterocyclyl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, 8-10-membered heteroaryl, phenyl, naphthyl, 5-membered or 6-membered heterocyclyl, 8-10-membered heterocyclyl, and the heteroaryl and heterocyclyl each independently contain 1-4 heteroatoms selected from N, O or S; the 5-membered heteroaryl, 8-10-membered heteroaryl, phenyl, naphthyl, 5-membered or 6-membered heterocyclyl, 8-10-membered heterocyclyl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, 8-10-membered heteroaryl, 5-membered or 6-membered heterocyclyl, 8-10-membered heterocyclyl, and the heteroaryl and heterocyclyl each independently contain 1-4 heteroatoms selected from N, O or S; and the 5-membered heteroaryl, 8-10-membered heteroaryl, 5-membered or 6-membered heterocyclyl, and 8-10-membered heterocyclyl are each independently substituted by one or more R d .
  • R 2b is selected from 5-membered heteroaryl, phenyl and 5- or 6-membered heteroaryl, 5- or 6-membered heteroaryl and 5- or 6-membered heteroaryl, 5- or 6-membered heterocyclyl, 5- or 6-membered heterocyclyl and 5- or 6-membered heterocyclyl; the heteroaryl and heterocyclyl each independently contain 1-2 heteroatoms selected from N, O or S; the 5-membered heteroaryl, phenyl and 5- or 6-membered heteroaryl, 5- or 6-membered heteroaryl and 5- or 6-membered heteroaryl, 5- or 6-membered heterocyclyl, 5- or 6-membered heterocyclyl and 5- or 6-membered heterocyclyl are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from 5-membered heteroaryl, phenyl and 6-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, 6-membered heterocyclyl, 6-membered heterocyclyl and pyridone, wherein the heteroaryl, heterocyclyl Each independently contains 1-2 heteroatoms selected from N, O or S; the 5-membered heteroaryl, phenyl-6-membered heteroaryl, 6-membered heteroaryl-5-membered heteroaryl, 6-membered heterocyclyl, 6-membered heterocyclyl-pyridinone group are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from pyrazolyl, thiazolyl, thienyl, isothiazolyl, phenylpyridinyl, pyridopyrrolyl, morpholinylpyridonyl, pyridonyl; and the pyrazolyl, thiazolyl, thienyl, isothiazolyl, phenylpyridinyl, pyridopyrrolyl, morpholinylpyridonyl, pyridonyl are each independently substituted with 1, 2, 3 or 4 R d .
  • each R d is independently selected from halogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, cyano, amino or hydroxyl.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 5-6 alkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, C 1-4 haloalkyl, C 5-6 haloalkyl, cyano or hydroxy.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 3-6 cycloalkyl, C 1-4 haloalkyl, cyano or hydroxy.
  • each Rd is independently selected from methyl, ethyl, propyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, trifluoromethyl, or hydroxy.
  • each R d is independently selected from methyl, Cl, Br, cyclopropyl, or hydroxy.
  • each R d is independently selected from C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, cyano, amino, hydroxyl or halogen, wherein at least one R d is hydroxyl.
  • each R d is independently selected from halogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, cyano or hydroxy, wherein at least one R d is hydroxy.
  • each R d is independently selected from halogen, C 1-6 alkyl, C 3-6 cycloalkyl, C 1-6 haloalkyl, cyano or hydroxy, wherein one and only one R d is hydroxy.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 5-6 alkyl, C 1-4 haloalkyl, C 5-6 haloalkyl, C 3-4 cycloalkyl, C 5-6 cycloalkyl, cyano or hydroxy, wherein one and only one R d is hydroxy.
  • each R d is independently selected from halogen, C 1-4 alkyl, C 1-4 haloalkyl, C 3-6 cycloalkyl, cyano or hydroxy, wherein one and only one R d is hydroxy.
  • each R d is independently selected from methyl, ethyl, propyl, F, Cl, Br, cyclopropyl, cyclobutyl, cyclopentyl, trifluoromethyl or hydroxy, wherein one and only one R d is hydroxy.
  • each R d is independently selected from methyl, Cl, Br, cyclopropyl, or hydroxy, wherein one and only one R d is hydroxy.
  • each R d is independently selected from methyl, Cl, Br, cyclopropyl or hydroxy, wherein one and only one R d is hydroxy, and the hydroxy is substituted at the ortho position to the site of attachment of R 2b to the main group.
  • R 2b is selected from Each independently may be further substituted with 1, 2 or 3 R d .
  • R 2b is selected from
  • R 2b is selected from 5-membered heteroaryl, each of which is independently substituted with 1, 2, 3 or 4 R d .
  • R 2b is selected from pyrazolyl, thiazolyl, thienyl, isothiazolyl, and said pyrazolyl, thiazolyl, thienyl, isothiazolyl are each independently substituted with 1, 2, 3 or 4 R d .
  • R 2b is selected from It may be further substituted with 1, 2 or 3 R d .
  • R 2b is selected from It may be further substituted with 1, 2 or 3 R d .
  • R 2b is selected from:
  • R 2b is selected from:
  • R 2b is selected from 6-membered heterocyclyl, 6-membered heterocyclyl and 6-membered heterocyclyl, wherein the heterocyclyl contains 1-2 heteroatoms selected from N, O or S; and the 6-membered heterocyclyl, 6-membered heterocyclyl and 6-membered heterocyclyl are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from morpholinylpyridonyl and pyridonyl; and the morpholinylpyridonyl and pyridonyl are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from pyridone; the pyridone is substituted by 1, 2, 3 or 4 R d substitutions.
  • R 2b is selected from 2-pyridonyl; said 2-pyridonyl is substituted with 1, 2, 3 or 4 R d .
  • R 2b is selected from It may be further substituted with 1, 2 or 3 R d .
  • R 2b is selected from It may be further substituted with 1, 2 or 3 R d .
  • R 2b is selected from
  • R 2b is selected from
  • R 2b is selected from phenyl and 5-membered or 6-membered heteroaryl, 5-membered or 6-membered heteroaryl and 5-membered or 6-membered heteroaryl; the heteroaryl contains 1-2 heteroatoms selected from N, O or S; the phenyl and 5-membered or 6-membered heteroaryl, 5-membered or 6-membered heteroaryl and 5-membered or 6-membered heteroaryl are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from phenyl and 6-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl, wherein the heteroaryl contains 1-2 heteroatoms selected from N, O or S; and the phenyl and 6-membered heteroaryl, 6-membered heteroaryl and 5-membered heteroaryl are each independently substituted by 1, 2, 3 or 4 R d .
  • R 2b is selected from phenylpyridinyl and pyridopyrrolyl; and the phenylpyridinyl and pyridopyrrolyl are each independently substituted with 1, 2, 3 or 4 R d .
  • R 2b is selected from It may be further substituted with 1, 2 or 3 R d .
  • R 2b is selected from
  • R1 is selected from halogen, C1-6 alkyl, 5-membered or 6-membered cycloalkenyl, 5-10-membered heterocyclyl, 5-membered or 6-membered heteroaryl, phenyl, the heterocyclyl and heteroaryl each independently contain 1-2 heteroatoms selected from N, O or S,
  • the alkyl, cycloalkenyl, heterocyclyl, heteroaryl and phenyl groups are each independently unsubstituted or substituted by one or more R g .
  • R1 is selected from a 5-membered or 6-membered cycloalkenyl, a 5-10-membered heterocyclyl, a 5-membered or 6-membered heteroaryl, or a phenyl group, wherein the heterocyclyl group and the heteroaryl group each independently contain 1-2 heteroatoms selected from N, O or S, and the cycloalkenyl group, the heterocyclyl group, the heteroaryl group, and the phenyl group are each independently unsubstituted or substituted by one or more Rg .
  • R1 is selected from 5-membered or 6-membered cycloalkenyl, 5-10-membered heterocycloalkyl, 5-membered or 6-membered heterocycloalkenyl, 5-membered or 6-membered heteroaryl, and phenyl, wherein the heterocycloalkyl, heterocycloalkenyl, and heteroaryl each independently contain 1-2 heteroatoms selected from N, O, or S, and the cycloalkenyl, heterocycloalkyl, heterocycloalkenyl, heteroaryl, and phenyl are each independently unsubstituted or substituted by one or more Rg .
  • R1 is selected from 6-membered cycloalkenyl, 6-9-membered heterocycloalkyl, 6-membered heterocycloalkenyl, pyridyl, pyrimidinyl, pyridazinyl, and the heterocycloalkyl and heterocycloalkenyl groups each independently contain 1-2 heteroatoms selected from N, O or S, and the cycloalkenyl, heterocycloalkyl and heterocycloalkenyl groups are each independently unsubstituted or substituted by one or more Rg .
  • R1 is selected from
  • R 1 is selected from
  • R 1 is selected from
  • R 1 is selected from halogen, C 1-6 alkyl, 5-membered or 6-membered heterocyclyl, wherein the heterocyclyl contains 1-2 heteroatoms selected from N, O or S, and the alkyl, heterocyclyl is unsubstituted or substituted by 1 or more R g .
  • R 1 is selected from a 5-membered or 6-membered heterocyclyl, wherein the heterocyclyl contains 1-2 heteroatoms selected from N, O or S, and the heterocyclyl is unsubstituted or substituted with one or more R g .
  • R 1 is selected from 5-membered or 6-membered heterocycloalkyl, 5-membered or 6-membered heterocycloalkenyl, wherein the heterocycloalkyl or heterocycloalkenyl contains 1-2 heteroatoms selected from N, O or S, and the heterocycloalkyl or heterocycloalkenyl is unsubstituted or substituted by 1 or more R g .
  • R 1 is selected from 5-membered or 6-membered heterocycloalkenyl, said heterocycloalkenyl containing 1-2 heteroatoms selected from N or O, said heterocycloalkenyl is unsubstituted or substituted with 1 or more R g .
  • R is selected from is unsubstituted or substituted with one or more Rg .
  • each Rg is independently selected from hydroxy, halogen, C1-4 alkyl, C1- 4 alkoxy; the C 1-4 alkyl and C 1-4 alkoxy are unsubstituted or substituted by one or more substituents selected from halogen, hydroxyl and C 1-2 alkoxy.
  • each Rg is independently selected from hydroxyl, halogen, C1-2 alkyl, C3-4 alkyl, C1-2 alkoxy, C3-4 alkoxy; the alkyl, alkoxy is unsubstituted or substituted with one or more substituents selected from halogen, hydroxyl, C1-2 alkoxy.
  • each Rg is independently selected from hydroxyl, F, Cl, C1-2 alkyl, C1-2 alkoxy; the alkyl, alkoxy is unsubstituted or substituted with one or more substituents selected from halogen, hydroxyl, C1-2 alkoxy.
  • each Rg is independently selected from hydroxyl, F, methyl, ethyl; the methyl, ethyl is unsubstituted or substituted with one or more substituents selected from halogen, hydroxyl, methoxy.
  • R is selected from
  • R is selected from
  • R is selected from
  • R 3 is selected from C 1-4 alkyl, which is unsubstituted or substituted with 1, 2 or 3 substituents selected from hydroxy or halogen.
  • R 3 is selected from C 1-4 alkyl, which is unsubstituted.
  • R 3 is selected from C 1-2 alkyl, which is unsubstituted.
  • R3 is selected from methyl or ethyl.
  • R3 is selected from ethyl.
  • R 4 is selected from H, halogen, C 1-4 alkyl, said alkyl being unsubstituted or substituted with 1, 2 or 3 halogens.
  • R 4 is selected from H, halogen, C 1-4 alkyl, which is unsubstituted or substituted with 1, 2 or 3 F.
  • R 4 is selected from H, F, Cl, Br, C 1-4 alkyl, and the C 1-4 alkyl is unsubstituted.
  • R 4 is selected from H, F, Cl, Br, C 1-2 alkyl, wherein the C 1-2 alkyl is unsubstituted or substituted with 1, 2 or 3 F.
  • R 4 is selected from H, F, Cl, Br, methyl, ethyl, and the methyl and ethyl groups are each independently unsubstituted or substituted with 1, 2 or 3 F groups.
  • R 4 is selected from H, F, Cl, Br, methyl, -CF 3 .
  • R 4 is selected from Cl, methyl, -CF 3 .
  • R 4 is selected from H, F, Cl, Br, methyl, ethyl, and the methyl and ethyl groups are unsubstituted.
  • R 4 is selected from H, F, Cl, Br or methyl.
  • R 4 is selected from Cl or methyl.
  • R 5 is selected from H, methyl, halogen.
  • R 5 is selected from H, F, Cl or methyl.
  • R 5 is selected from H, Cl or methyl.
  • R5 is selected from H.
  • R 6 is selected from H, halogen, C 1-4 alkyl; the alkyl is unsubstituted or substituted with 1, 2 or 3 halogens.
  • R 6 is selected from H, halogen, C 1-4 alkyl, which is unsubstituted or substituted with 1, 2 or 3 F.
  • R 6 is selected from H, halogen, C 1-2 alkyl, which is unsubstituted or substituted with 1, 2 or 3 F.
  • R 6 is selected from H, F, Cl, methyl, ethyl, wherein the methyl, ethyl is unsubstituted or substituted with 1, 2 or 3 F.
  • R 6 is selected from H, F, Cl, methyl, or -CF 3 .
  • R 6 is selected from H, F or Cl.
  • R 6 is selected from H, halogen, C 1-4 alkyl, wherein the alkyl is substituted with 1, 2 or 3 F.
  • R 6 is selected from H, halogen, C 1-2 alkyl, wherein the alkyl is substituted with 1, 2 or 3 F.
  • R 6 is selected from H, F, Cl, methyl, ethyl, and the methyl, ethyl is substituted with 1, 2 or 3 F.
  • R 6 is selected from H, F, Cl or -CF 3 .
  • R 6 is selected from H, F or Cl.
  • R 7 is selected from H, -SF 5 , -C(O)H, halogen, C 1-4 alkyl, C 2-4 alkenyl, C 2-4 alkynyl, C 3-5 cycloalkyl; the alkyl and cycloalkyl are unsubstituted or substituted with one or more halogens.
  • R 7 is selected from -SF 5 , -C(O)H, halogen, C 1-4 alkyl; said alkyl is unsubstituted or substituted with 1, 2 or 3 halogens.
  • R 7 is selected from -SF 5 , -C(O)H, halogen, C 1-4 alkyl; said alkyl is unsubstituted or substituted with 1, 2 or 3 Fs.
  • R 7 is selected from -SF 5 , -C(O)H, halogen, C 1-2 alkyl; said alkyl is unsubstituted or substituted with 1, 2 or 3 Fs.
  • R 7 is selected from -SF 5 , -C(O)H, F, Cl, Br, methyl, ethyl; the methyl, ethyl is unsubstituted or substituted with 1, 2 or 3 Fs.
  • R7 is selected from -CF3 , -CHF2 , -CH2CH3 , Cl, Br, -SF5 , or -C(O)H.
  • R 7 is selected from -CF 3 .
  • R 7 is selected from -OCF 3 .
  • R 8 is selected from H or F.
  • formula (I) has a general structure selected from formula (IB):
  • R 1 , R 2b , R 3 and R 4 are each as defined in formula (I).
  • formula (I) has the structure shown in formula (IB-1):
  • R 1 , R 2b , and R 4 are as defined in formula (I).
  • formula (I) has a structure selected from formula (IIA):
  • R 1 , R 2b , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • formula (IIA) has the structure shown in formula (IIA-1):
  • R 1 , R d , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • the formula (IIA-1) has the structure shown in formula (IIA-1-1):
  • R 1 and R d are as defined in formula (I).
  • formula (IIA) has the structure shown in formula (IIA-2):
  • R 1 , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • the formula (IIA-2) has the structure shown in formula (IIA-2-1):
  • R 1 is as defined in formula (I).
  • formula (IIA) has the structure shown in formula (IIA-3):
  • R 1 , R d , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • the formula (IIA-3) has the structure shown in formula (IIA-3-1):
  • R 1 and R d are as defined in formula (I).
  • formula (IIA) has the structure shown in formula (IIA-4):
  • ring B is a 5-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N, O and S; the 5-membered heteroaryl group is unsubstituted or substituted by 1 or more C 1-4 alkyl groups; X, R 4 , R 5 , R 6 and R 7 are defined as above.
  • Ring B is a 5-membered heteroaryl; the 5-membered heteroaryl contains 1 heteroatom selected from N, O and S; the 5-membered heteroaryl is unsubstituted or substituted with 1 or more C 1-4 alkyl groups; X, R 4 , R 5 , R 6 , and R 7 are as defined above.
  • Ring B is selected from furanyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl; the furanyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl are each independently unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; X, R 4 , R 5 , R 6 , R 7 are as defined above.
  • Ring B is selected from furanyl, pyrrolyl, thienyl; the furanyl, pyrrolyl, thienyl are each independently unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; X, R 4 , R 5 , R 6 , R 7 are as defined above.
  • Ring B is selected from furanyl and pyrrolyl; the furanyl and pyrrolyl are each independently unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl; X, R 4 , R 5 , R 6 and R 7 are as defined above.
  • ring B is selected from furanyl, pyrrolyl, thienyl; the pyrrolyl is unsubstituted or substituted with one or more selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, The substituents of the isobutyl group and the tert-butyl group are substituted; X, R 4 , R 5 , R 6 and R 7 are as defined above.
  • Ring B is selected from furanyl and pyrrolyl; the pyrrolyl is unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl; X, R 4 , R 5 , R 6 and R 7 are as defined above.
  • Ring B is selected from furanyl, pyrrolyl, thienyl; the pyrrolyl is unsubstituted or substituted with 1 methyl group; X, R 4 , R 5 , R 6 , and R 7 are as defined above.
  • Ring B is selected from furanyl and pyrrolyl; the pyrrolyl is unsubstituted or substituted with 1 methyl group; X, R 4 , R 5 , R 6 , and R 7 are as defined above.
  • formula (I) has a structure selected from formula (IIB):
  • R 1 , R 2b , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • formula (IIB) has the structure shown in formula (IIB-1):
  • R 1 , R d , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • formula (IIB-1) has the structure shown in formula (IIB-1-1):
  • R 1 and R d are as defined in formula (I).
  • formula (IIB) has the structure shown in formula (IIB-2):
  • R 1 , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • the formula (IIB-2) has the structure shown in formula (IIB-2-1):
  • R 1 is as defined in formula (I).
  • formula (IIB) has the structure shown in formula (IIB-3):
  • R 1 , R d , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • the formula (IIB-3) has the structure shown in formula (IIB-3-1):
  • R 1 and R d are as defined in formula (I).
  • the formula (IIB) has a structure shown in formula (IIB-4):
  • ring B is a 5-membered heteroaryl group; the 5-membered heteroaryl group contains 1-2 heteroatoms selected from N, O and S; the 5-membered heteroaryl group is unsubstituted or substituted by 1 or more C 1-4 alkyl groups; X, R 4 , R 5 , R 6 and R 7 are defined as above.
  • Ring B is a 5-membered heteroaryl; the 5-membered heteroaryl contains 1 heteroatom selected from N, O and S; the 5-membered heteroaryl is unsubstituted or substituted with 1 or more C 1-4 alkyl groups; X, R 4 , R 5 , R 6 , and R 7 are as defined above.
  • Ring B is selected from furanyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl; the furanyl, pyrrolyl, thienyl, pyrazolyl, imidazolyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl are each independently unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; X, R 4 , R 5 , R 6 , R 7 are as defined above.
  • Ring B is selected from furanyl, pyrrolyl, thienyl; the furanyl, pyrrolyl, thienyl are each independently unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; X, R 4 , R 5 , R 6 , R 7 are as defined above.
  • Ring B is selected from furanyl and pyrrolyl; the furanyl and pyrrolyl are each independently unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl; X, R 4 , R 5 , R 6 and R 7 are as defined above.
  • Ring B is selected from furanyl, pyrrolyl, thienyl; the pyrrolyl is unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl; X, R 4 , R 5 , R 6 , R 7 are as defined above.
  • Ring B is selected from furanyl and pyrrolyl; the pyrrolyl is unsubstituted or substituted with one or more substituents selected from methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl and tert-butyl; X, R 4 , R 5 , R 6 and R 7 are as defined above.
  • Ring B is selected from furanyl, pyrrolyl, thienyl; the pyrrolyl is unsubstituted or substituted with 1 methyl group; X, R 4 , R 5 , R 6 , and R 7 are as defined above.
  • Ring B is selected from furanyl and pyrrolyl; the pyrrolyl is unsubstituted or substituted with 1 methyl group; X, R 4 , R 5 , R 6 , and R 7 are as defined above.
  • formula (I) has a structure selected from formula (IIC):
  • R 1 , R 2b , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • formula (IIC) has the structure shown in formula (IIC-1):
  • R 1 , R d , R 4 , R 5 , R 6 and R 7 are as defined in formula (I).
  • the formula (IIC-1) has the structure shown in formula (IIC-1-1):
  • R 1 and R d are as defined in formula (I).
  • formula (IIC) has the structure shown in formula (IIC-2):
  • R 1 , R 4 , R 5 , R 6 , and R 7 are defined as follows The definitions given in formula (I).
  • the formula (IIC-2) has the structure shown in formula (IIC-2-1):
  • R 1 is as defined in formula (I).
  • formula (I) is selected from the following specific compounds:
  • pharmaceutically acceptable salt refers to a salt of a compound of the present invention, prepared from a compound having a specific substituent discovered by the present invention and a relatively non-toxic acid or base.
  • a base addition salt can be obtained by contacting such compound with a sufficient amount of a base in a pure solution or a suitable inert solvent.
  • an acid addition salt can be obtained by contacting such compound with a sufficient amount of an acid in a pure solution or a suitable inert solvent.
  • prodrug refers to derivatives of the compounds of formula (I) with specific substituents discovered in the present invention, which may have weak activity or even no activity themselves, but after administration, they are converted into compounds with specific substituents discovered in the present invention under physiological conditions (for example, by metabolism, solvent decomposition or other means), and produce corresponding biological activity in vivo.
  • the term "metabolite” refers to the product obtained by the metabolism of the compound of formula (I) with specific substituents found in the present invention in vivo.
  • the metabolite of a compound can be identified by techniques known in the art, and its activity can be characterized by the test methods described in the present invention. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, amidation, deamidation, esterification, defatting, enzymatic cleavage, etc.
  • the present invention includes the metabolites of the compound, including the metabolites produced by contacting the compound of the present invention with a mammal for a period of time.
  • deuterated compound refers to a compound of the invention that includes at least one deuterium atom, specifically one or more hydrogen atoms in the compound of the invention may be replaced or substituted with a deuterium atom.
  • the compound includes two or more deuterium atoms.
  • the compound includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Methods for the synthesis of organic compounds are known in the art.
  • the present invention also provides a method for preparing the compound.
  • the preparation of the compound described in the general formula (I) of the present invention can be completed by the following illustrative methods and examples, but these methods and examples should not be considered in any way to limit the scope of the present invention.
  • the compound described in the present invention can also be synthesized by synthetic techniques known to those skilled in the art, or a combination of synthetic methods known in the art and the method described in the present invention.
  • the product obtained by each step of the reaction is obtained using separation techniques known in the art, including but not limited to extraction, filtration, distillation, crystallization, chromatographic separation, etc.
  • the starting materials and chemical reagents required for the synthesis can be conventionally synthesized according to the literature (such as provided by Scifinder) or purchased.
  • the compound of the general formula (I) of the present invention can be synthesized according to the following method:
  • W represents halogen
  • PG is a protecting group such as -Boc (tert-butyloxycarbonyl), benzyl, PMB, and R1 , R3 , R4 , R5 , R6 , R7 , X, R2a and R2b are defined as described in formula (I).
  • W represents halogen
  • PG is a protecting group such as Boc (tert-butyloxycarbonyl), benzyl, PMB, and R1 , R3 , L, R4 , R5 , R6 , R7 , X, R2a , R2b and R2e are defined as described in formula (I).
  • the present invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising the above-mentioned compound, or its stereoisomer, tautomer, polymorph, cocrystal, solvate, metabolite, prodrug, deuterated compound, pharmaceutically acceptable salt, and optional pharmaceutical excipients.
  • the pharmaceutical excipient is preferably a pharmaceutically acceptable carrier, diluent, excipient or a combination thereof.
  • the present invention also provides a pharmaceutical composition for treating and/or preventing diseases related to the biological activity of WRN, the pharmaceutical composition comprising a therapeutic and/or preventive effective amount of the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, and optional pharmaceutical excipients.
  • the pharmaceutical excipient is preferably a pharmaceutically acceptable carrier, diluent, excipient or a combination thereof.
  • the pharmaceutical composition may further comprise other drugs for treating and/or preventing diseases associated with the biological activity of WRN.
  • the disease associated with the biological activity of WRN is a tumor or cancer.
  • the disease associated with WRN biological activity is an MSI tumor or an MSI cancer.
  • the present invention also provides a WRN inhibitor, which comprises a therapeutically and/or preventively effective amount of the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, solvates, metabolites, prodrugs, deuterated compounds, and pharmaceutically acceptable salts.
  • the present invention also provides the use of the aforementioned compound, or its stereoisomers, tautomers, polymorphs, cocrystals, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the aforementioned pharmaceutical compositions in the preparation of WRN inhibitors.
  • the present invention also provides the use of the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the pharmaceutical composition as described above in the preparation of drugs for treating and/or preventing diseases related to the biological activity of WRN.
  • the present invention also provides the use of the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the pharmaceutical composition as described above in the treatment and/or prevention of diseases related to the biological activity of WRN.
  • the present invention also provides the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the pharmaceutical composition as described above, which is used to treat and/or prevent diseases related to the biological activity of WRN.
  • the above-mentioned disease associated with the biological activity of WRN is a tumor or cancer.
  • the above-mentioned disease associated with the biological activity of WRN is an MSI tumor or an MSI cancer.
  • the present invention also provides a method for treating and/or preventing diseases associated with the biological activity of WRN, comprising administering to an individual in need thereof a therapeutically and/or preventively effective amount of the compound as described above, or its stereoisomers, tautomers, polymorphs, cocrystals, solvates, metabolites, prodrugs, deuterated compounds, pharmaceutically acceptable salts, or the pharmaceutical composition as described above.
  • the above-mentioned disease associated with the biological activity of WRN is a tumor or cancer.
  • the above-mentioned disease associated with the biological activity of WRN is an MSI tumor or an MSI cancer.
  • treatment generally refers to obtaining the desired pharmacological and/or physiological effect.
  • the effect can be preventive, in terms of completely or partially preventing a disease or its symptoms; and/or therapeutic, in terms of partially or completely stabilizing or curing a disease and/or side effects caused by a disease.
  • treatment encompasses any treatment of a patient's disease, including: (a) preventing a disease or symptom from occurring in a patient who is susceptible to the disease or symptom but has not yet been diagnosed with the disease; (b) inhibiting the symptoms of a disease, i.e., preventing its development; or (c) alleviating the symptoms of a disease, i.e., causing the disease or symptom to regress.
  • a vertebrate refers to a mammal.
  • Mammals include, but are not limited to, livestock (such as cattle), pets (such as cats, dogs, and horses), primates, mice, and rats.
  • a mammal refers to a human.
  • an effective amount refers to an amount effective to achieve the desired therapeutic or preventive effect at the necessary dosage and time.
  • the "therapeutically effective amount” of the substance/molecule of the present invention may vary according to factors such as the disease state, age, sex and weight of the individual and the ability of the substance/molecule to elicit the desired response in the individual.
  • An effective amount also encompasses an amount in which the therapeutically beneficial effects of the substance/molecule outweigh any toxic or deleterious consequences.
  • a “prophylactically effective amount” refers to an amount that is effective at the necessary dose and time to achieve the desired prophylactic effect.
  • a therapeutically effective amount of a drug can reduce the number of cancer cells; reduce tumor volume; inhibit (i.e., slow down to a certain extent, preferably stop) cancer cell infiltration into surrounding organs; inhibit (i.e., slow down to a certain extent, preferably stop) tumor metastasis; inhibit tumor growth to a certain extent; and/or alleviate one or more symptoms associated with cancer to a certain extent.
  • C3-6cycloalkyl- ( C1-6alkyl ) r- means that it is attached to the rest of the molecule through ( C1 - C6alkyl ) r- .
  • substituted means that any one or more hydrogens on a designated atom or group are replaced with the selection of a designated group, provided that the normal valency of the designated atom is not exceeded.
  • C 1-6 alkyl specifically refers to the independently disclosed methyl, ethyl, C 3 alkyl, C 4 alkyl, C 5 alkyl and C 6 alkyl, or the independently disclosed "C 1-4 alkyl", or the independently disclosed "C 1-3 alkyl”.
  • alkyl refers to a branched and straight chain saturated aliphatic hydrocarbon group including a specified number of carbon atoms.
  • C 1-6 alkyl refers to C 1 , C 2 , C 3 , C 4 , C 5 and C 6.
  • C 1-6 alkyl refers to an alkyl group having 1 to 6 carbon atoms.
  • the alkyl group may be unsubstituted or substituted so that one or more of its hydrogens are replaced by another chemical group.
  • alkyl groups include, but are not limited to, methyl, ethyl, propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, tert-butyl), pentyl (e.g., n-pentyl, isopentyl, neopentyl), etc.
  • C 1-6 alkyl herein includes monovalent C 1-6 alkyl and divalent C 1-6 alkylene, for example, C 3 -C 6 cycloalkyl-(C 1 -C 6 alkyl) r - in which C 1 -C 6 alkyl refers to C 1 -C 6 alkylene.
  • alkoxy refers to any of the above alkyl groups (eg, C 1-6 alkyl, C 1-4 alkyl, C 1-3 alkyl , etc.) which is attached to the rest of the molecule through an oxygen atom (-O-).
  • C 1-6 haloalkyl or “C 1-6 haloalkoxy” refers to an alkyl or alkoxy group in which one or more (e.g., 2 or 3) hydrogen atoms are replaced by halogen atoms, such as fluorine, chlorine, or bromine.
  • the alkyl or alkoxy group is as defined above.
  • the term "halogenated C 1-6 alkyl” is preferably fluorinated, such as -CF 3 , -CHF 2 , -CH 2 F, -CH 2 CH 2 F, -CH 2 CHF 2 , -CH 2 CF 3 , and the like.
  • halogenated C 1-6 alkoxy is preferably fluorinated, such as -OCF 3 , -OCHF 2 , -OCH 2 F, -OCH 2 CH 2 F, -OCH 2 CHF 2 , -OCH 2 CF 3 , and the like.
  • hydroxy-substituted C 1-6 alkyl means that one of the hydrogen atoms in the alkyl group is replaced by a hydroxy group.
  • the definition of the alkyl group is as described above.
  • the "hydroxy-substituted C 1-6 alkyl group” may be a hydroxymethyl group.
  • alkenyl refers to a hydrocarbon group including a straight or branched configuration and having one or more carbon-carbon double bonds that may be present at any stable point along the chain.
  • C2-6 alkenyl refers to groups including C2 , C3 , C4 , C5 , and C6 .
  • alkenyl examples include, but are not limited to, vinyl, 1-propenyl, 2-propenyl, 2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl, 4-methyl-3-pentenyl, and the like.
  • cycloalkyl refers to a cyclized alkyl group, including a monocyclic, bicyclic or polycyclic system.
  • each ring should be a saturated carbocyclic ring or a carbocyclic residue, and the connection between each two rings of the bicyclic or polycyclic cycloalkyl group includes bridging, fusion or spiro connection.
  • C 3-10 cycloalkyl refers to C 3 , C 4 , C 5 , C 6 , C 7 , C 8 , C 9 and C 10 cycloalkyl.
  • Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wait.
  • cycloalkenyl refers to a carbocyclic group having at least one carbon-carbon double bond in a cycloalkyl group as defined above, for example
  • carrier or “carbocycle residue” refers to any stable 3-, 4-, 5-, 6- or 7-membered monocyclic ring or 5-, 6-, 7-, 8-, 9-, 10-, 11-, 12-, 13-, 14-membered bicyclic or polycyclic ring, any one of which may be saturated, partially saturated, unsaturated or aromatic.
  • the bicyclic or polycyclic carbocycles may be connected in a manner that includes bridging, fusion or spiro connection.
  • carbocycles include, but are not limited to, cyclopropyl, cyclobutyl, cyclobutenyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptenyl, cycloheptyl, adamantyl, cyclooctyl, phenyl, naphthyl, [2,2,2]bicyclooctane, wait.
  • aryl refers to a monocyclic, bicyclic or tricyclic aromatic hydrocarbon group having 5 to 14 carbon atoms in the ring portion.
  • each of its rings is an aromatic ring.
  • the connection modes of each two rings of the bicyclic or tricyclic aryl group include bridging, fusion, and spiro connection. For example, phenyl and naphthyl, each of which can be substituted.
  • heterocycle refers to substituted and unsubstituted 4-8 membered monocyclic or bicyclic groups, 8-10 membered bicyclic or tricyclic groups and 10-14 membered tricyclic or polycyclic groups, wherein at least one ring has at least one heteroatom (O, S or N), and the heteroatom-containing ring preferably has 1, 2 or 3 heteroatoms selected from O, S and N.
  • Each heteroatom-containing ring in the group may contain 1 or 2 oxygen or sulfur atoms and/or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less, and further provided that the ring contains at least one carbon atom.
  • the heteroatom refers only to N or O, and the total number does not exceed 3, preferably only 1-2 heteroatoms.
  • the fused rings completing the bicyclic and tricyclic groups may contain only carbon atoms and may be saturated, partially saturated or completely unsaturated, aromatic or non-aromatic.
  • the heterocyclic group may be attached on any available nitrogen or carbon atom.
  • the heterocyclic group herein, the fused rings completing the bicyclic and tricyclic groups are non-aromatic rings.
  • heterocyclic groups include "heterocycloalkyl” and "heterocycloalkenyl” as described below.
  • heterocyclic groups include, but are not limited to, azetidinyl, oxetanyl, pyrrolidinyl, imidazolinyl, oxazolidinyl, isoxazolinyl, thiazolidinyl, tetrahydrofuranyl, piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazacycloheptatrienyl, 1-pyridonyl, 4-piperidonyl, tetrahydropyranyl, morpholinyl, 1,3-dioxolane, quinuclidine, wait.
  • heterocycloalkyl refers to a heterocyclic group in which all the ring atoms are fully saturated, as defined above, for example When it is bicyclic or tricyclic, “heterocycloalkyl” includes “heterobridged cycloalkyl”, “heterospirocycloalkyl” and “heteroparacycloalkyl”.
  • heterocycloalkenyl refers to a heterocyclic group having at least one carbon-carbon double bond in the heterocyclic ring as defined above, for example
  • the heterocycloalkenyl group includes “heterobridged cycloalkenyl group", “heterospirocycloalkenyl group” and “heterocycloalkenyl group”.
  • the heterocycloalkenyl group is bicyclic or tricyclic, the ring is unsaturated as a whole and cannot form aromaticity.
  • heteroaryl refers to substituted and unsubstituted aryl groups described above having at least one heteroatom (O, N or S) in at least one ring, including aromatic 5-8-membered monocyclic groups, 8-10-membered bicyclic groups and 10-14-membered tricyclic groups, and the heteroatom-containing ring preferably has 1, 2 or 3 heteroatoms selected from O, N or S.
  • Each heteroatom-containing ring of the heteroaryl group may contain 1 or 2 oxygen or sulfur atoms and/or 1 to 4 nitrogen atoms, provided that the total number of heteroatoms in each ring is 4 or less and each ring has at least one carbon atom.
  • Each ring of a bicyclic or tricyclic heteroaryl group is an aromatic ring.
  • spirocyclyl refers to a bicyclic structure having one common ring atom, and each monocyclic ring is a saturated or unsaturated, aromatic or non-aromatic carbocyclic ring having 3-7 carbon atoms.
  • exemplary spirocyclyls include, but are not limited to, spiro[4.5]decane, spiro[3.4]octane, spiro[2.3]hexane,
  • the spirocyclic group does not include the aryl group defined above.
  • spiro heterocyclyl refers to a bicyclic structure with one common ring atom, and each monocyclic ring is a saturated or unsaturated monocyclic group with 3-8 ring atoms, wherein at least one ring has 1 or 2 ring atoms selected from N, O or S (O) n heteroatoms, wherein n is an integer from 0 to 2, and the remaining ring atoms are C.
  • 1 or 2 ring carbon atoms in the heterocyclyl ring can be optionally replaced by -CO- groups.
  • Exemplary spiro heterocyclyls include, but are not limited to: 5-azaspiro [2.3] hexane, 6-oxaspiro [3.4] -7-octanone.
  • the spiro heterocyclyl does not include heteroaryl as defined above.
  • heteroatom shall include oxygen, sulfur and nitrogen.
  • halogen shall include "F, Cl, Br, I".
  • saturated When the term “saturated” is used herein to refer to a ring or a group, unless otherwise specified, the ring or group shall be fully saturated.
  • C 1-6 alkyl optionally substituted with 1 to 3 R d means that the C 1-6 alkyl may be substituted with 1 to 3 R d or may not be substituted with 1 to 3 R d .
  • Other similar definitions can be understood with reference to the above content.
  • XXX is substituted with one or more substituents selected from YYY
  • XXX may be substituted with one or more substituents at any substitutable position, and the substituents are selected from YYY.
  • the multiple substituents may be the same or different. Wherein, multiple is 2 or more, preferably 2, 3 or 4, more preferably 2 or 3.
  • C 1-6 alkyl is substituted with one or more substituents selected from cyano and hydroxyl at any substitutable position, which means that C 1-6 alkyl may be substituted with one or more cyano at any substitutable position, may be substituted with one or more hydroxyl at any substitutable position, and may be substituted with one or more cyano and hydroxyl (e.g., one cyano and one hydroxyl, or two cyano and one hydroxyl, or two cyano and two hydroxyl, etc.) at any substitutable position.
  • substituents selected from cyano and hydroxyl at any substitutable position which means that C 1-6 alkyl may be substituted with one or more cyano at any substitutable position, may be substituted with one or more hydroxyl at any substitutable position, and may be substituted with one or more cyano and hydroxyl (e.g., one cyano and one hydroxyl, or two cyano and one hydroxyl, or two cyano and two hydroxyl, etc
  • one or more H atoms in XXX can be further replaced by one or more identical or different YYY"
  • the H atoms in XXX can be replaced by YYY or not; the H atoms in XXX can be replaced by one or more identical YYY or one or more different YYY, and each substituent should not have the same sign and interfere with each other.
  • the compound represented by formula (I) of the present invention has a good WRN inhibitory effect and can be used as a drug for the treatment and/or prevention of diseases related to this effect, and can be particularly used for the treatment and/or prevention of MSI tumor-related diseases.
  • NMR nuclear magnetic resonance
  • MS mass spectrometry
  • Reaction monitoring and MS measurements were performed using a Thermofisher ESQ (ESI) mass spectrometer.
  • ESI Thermofisher ESQ
  • HPLC determination used a Thermo Fisher U3000DAD high pressure liquid chromatograph (GL Sciences ODS-HL HP 3 ⁇ m 3.0*100mm column).
  • the thin layer chromatography silica gel plate used was Qingdao Ocean GF254 silica gel plate.
  • the silica gel plate used in thin layer chromatography (TLC) was of 0.15-0.2 mm in size.
  • the thin layer chromatography separation and purification product used a high-performance thin layer chromatography preparative plate of 0.9-1.0 mm in size.
  • Column chromatography used Qingdao Ocean 200-300 mesh silica gel as a carrier.
  • the developing solvent used was A: dichloromethane and methanol system; B: petroleum ether and ethyl acetate system. The volume ratio of the solvent was adjusted according to the polarity of the compound.
  • the medium pressure preparative liquid phase purification used a biotage isera one preparative liquid phase.
  • the preparative liquid chromatograph (prep-HPLC) model was Agilent 1290 Infinity 2nd generation.
  • reaction raw materials can be purchased from manufacturers such as San Chemical Technology (Shanghai) Co., Ltd., Shanghai Shaoyuan Reagent Co., Ltd., Nanjing Yaoshi Technology Co., Ltd., Jiangsu Aikon Biopharmaceutical R&D Co., Ltd., and Shanghai Bid Pharmaceutical Technology Co., Ltd.
  • Step 1 Dissolve methyl 3-oxopentanoate (int-1a, 70 g, 538.05 mmol) in DCM (500 mL), add NBS (100.6 g, 564.95 mmol), then add TSOH.H 2 O (20.4 g, 107.61 mmol), and stir at room temperature for 3 hours. Filter by suction, and concentrate the filtrate under reduced pressure to obtain crude methyl 2-bromo-3-oxopentanoate (int-1b, 126.5 g, 602.87 mmol). MS Calcd: 207.97; MS Found: 206.98 ([MH] - ).
  • Step 2 Dissolve methyl 2-bromo-3-oxopentanoate (int-1b, 10 g, 47.85 mmol), tert-butyl piperazine-1-carboxylate (int-1c, 49.0 g, 263.16 mmol), and potassium carbonate (39.7 g, 287.08 mmol) in MeCN (100 mL) and stir at room temperature for 30 minutes.
  • Step 3 At room temperature, H 3 PO 4 (4.5 g, 46.01 mmol) was added to a solution of 5-bromo-2H-1,2,4-triazol-3-amine (int-1e, 5 g, 30.67 mmol) and tert-butyl 4-(1-methoxy-1,3-dioxopentan-2-yl)piperazine-1-carboxylate (int-1d, 10.6 g, 33.74 mmol) in anhydrous ethanol (50 mL), and heated to 80° C. with stirring for two days. Saturated sodium bicarbonate was added to quench the reaction, and the pH was adjusted to 6-7.
  • H 3 PO 4 4.5 g, 46.01 mmol
  • Step 1 Into a 50 mL eggplant-shaped bottle, tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-1, 400 mg, 0.94 mmol) and solvent 1,4-dioxane (5 mL) were added. Then, N-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (374 mg, 1.03 mmol) and DIPEA (383 mg, 2.81 mmol) were added thereto, and the mixture was stirred at room temperature at 85°C for 5 hours.
  • Step 2 tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2a, 350 mg, 0.53 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (166 mg, 0.79 mmol) and solvent 1,4-dioxane (3 mL), H 2 O (1.5 mL) were added to a 50 mL eggplant-shaped bottle.
  • Step 3 To a 50 mL eggplant shaped bottle, tert-butyl 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2b, 220 mg, 0.33 mmol) and solvent DCM (5 mL) were added. Then trifluoroacetic acid (1 mL) was added, and the mixture was stirred at room temperature overnight.
  • Step 1 Add 5-bromo-2H-1,2,4-triazole-3-amine (int-1e, 2.0 g, 12.3 mmol) and methyl 3-oxopentanoate (int-1a, 2.2 g, 15.3 mmol) to a 25 mL round-bottom flask, add AcOH (7 mL), heat to 80 ° C and stir overnight. After cooling to room temperature, stir at 0 ° C for 1 h, filter, and use a small amount of EtOH (10 mL) to remove the filter cake.
  • Step 2 Add 2-bromo-5-ethyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (int-3a, 1.63 g, 6.71 mmol) and N-[2-chloro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (2.67 g, 7.35 mmol) to a 25 mL round-bottom flask, add 1,4-dioxane (7 mL), and stir at 80 °C for 4 h under nitrogen protection. After cooling to room temperature, a white precipitate was precipitated, filtered, and the filtrate was concentrated.
  • Step 3 2-(2-bromo-5-ethyl-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-4-yl)-N-[2-chloro-4-(trifluoromethyl)phenyl]acetamide (int-3b, 3.0 g, 6.27 mmol) and 3,6-dihydro-2H-pyran-4-boronic acid pyranoside (1.71 g, 8.14 mmol) were added to a 100 mL two-necked bottle, and potassium phosphate (3.99 g, 18.80 mmol), Pd(dppf)Cl 2 (458 mg, 0.63 mmol), 1,4-dioxane (40 mL) and purified water (20 ml) were added in sequence.
  • Step 4 Add N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-3c, 1.35 g, 2.80 mmol) into a 50 mL flask, add NBS (548 mg) and acetonitrile (50 mL) in sequence, heat to 80 °C and continue the reaction for 6 h.
  • NBS 548 mg
  • acetonitrile 50 mL
  • Step 1 Add 4-bromo-2-fluoro-1-(trifluoromethyl)benzene (int-4a, 9.5 g, 39.09 mmol) and solvent THF (20 mL) were added and cooled to -78°C.
  • LDA 4 g, 43.00 mmol
  • iodomethane 4.76 g, 76 mL, 58.64 mmol
  • Saturated ammonium chloride was added to quench the reaction under ice bath, and the mixture was extracted with dichloromethane (50 mL*3).
  • Step 2 Add 1-bromo-3-fluoro-2-methyl-4-(trifluoromethyl)benzene (int-4b, 9.0 g, 35.02 mmol), tert-butyl carbamate (4.5 g, 38.52 mmol) and solvent 1,4-dioxane (20 mL) into a 250 mL eggplant-shaped bottle, add Xantphos (1.4 g, 2.45 mmol), Pd 2 (dba) 3 (1.6 g, 1.75 mmol), Cs 2 CO 3 (17.1 g, 52.53 mmol), and heat to 85°C under nitrogen protection for 3 hours. The mixture was cooled to room temperature, filtered, and the filter cake was washed with dichloromethane.
  • Step 3 Add tert-butyl 3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)carbamate (int-4c, 7.26 g, 24.75 mmol) and methanol solution of hydrochloric acid (20 mL) to a 250 mL eggplant-shaped bottle and react at room temperature for two hours. Add sodium hydroxide to the reaction solution to adjust the pH to neutral, filter out insoluble matter, wash the organic phase with 2*30 mL of water, dry the organic phase with anhydrous sodium sulfate, and concentrate to obtain a crude product: 3-fluoro-2-methyl-4-trifluoromethylaniline (int-4d, 4.3 g, 22.27 mmol, 89.6% yield). MS Calcd: 193.05; MS Found: 192.11 ([MH] - ).
  • Step 4 Add 3-fluoro-2-methyl-4-trifluoromethylaniline (int-4d, 4.3g, 22.27mmol), dichloromethane (30mL), triethylamine (6.17mL, 44.54mmol) to a 100mL eggplant-shaped bottle, add chloroacetyl chloride (2.65mL, 26.72mmol) in dichloromethane (10mL) dropwise under ice bath, then naturally return to room temperature to react overnight.
  • Step 5 Add 2-chloro-N-[3-fluoro-2-methyl-4-trifluoromethylphenyl]acetamide (int-4e, 5.5 g, 18.77 mmol), potassium iodide (6.8 g, 40.80 mmol) and solvent acetone (30 mL) to a 250 mL eggplant-shaped bottle, and then react at 50°C for 3 hours.
  • Step 1 tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-1,14 g, 32.8 mmol) was placed in a 250 mL three-necked flask containing dioxane (150 mL) and water (50 mL), followed by the addition of 3,6-dihydro-2H-pyran-4-boronic acid pyranoside (10.3 g, 49.1 mmol), K 2 CO 3 (9.1 g, 65.6 mmol), Pd(dppf)cl 2 (1.2 g, 1.64 mmol), replaced three times under nitrogen atmosphere, heated to 100 ° C for 14 hours, cooled to room temperature, added 300 mL of water, extracted three times with 300 mL of dichloromethane, the organic phase was washed with saturated brine,
  • Step 1 Dissolve ethyl 3-oxopentanoate (int-6a, 100.00 g, 693.63 mmol) in dichloromethane (1 L) in a 2L three-necked flask, and add NBS (129.62 g, 728.31 mmol) and TsOH*H 2 O (26.39 g, 138.73 mmol) in sequence. The resulting reaction mixture was stirred overnight at room temperature. After the reaction was completed, the reaction mixture was poured into saturated saline water (1 L) and extracted twice with dichloromethane (1 L*2). The combined organic phase was backwashed twice with brine, dried over anhydrous sodium sulfate, filtered, and spin-dried.
  • NBS 129.62 g, 728.31 mmol
  • TsOH*H 2 O 26.39 g, 138.73 mmol
  • Step 2 Dissolve ethyl 2-bromo-3-oxopentanoate (int-6b, 60.00 g, 268.98 mmol) in acetonitrile (300 mL) in a 1L single-mouth bottle, and add K 2 CO 3 (74.36 g, 537.96 mmol) and (R)-1-N-Boc-2-methylpiperazine (56.56 g, 282.43 mmol) in sequence. The resulting reaction mixture was stirred at room temperature overnight. The undissolved inorganic salts were filtered off, and the filtrate was concentrated to dryness.
  • Step 3 Weigh (2R)-4-(1-ethoxy-1,3-dioxopentan-2-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (int-6c, 91.32 g, 266.68 mmol) and place it in a 1L single-mouth bottle, add ethanol (150 mL) to dissolve, then add 5-bromo-2H-1,2,4-triazole-3-amine (int-1e, 47.81 g, 293.35 mmol) and phosphoric acid (27.44 g, 280.01 mmol) in sequence, switch the entire reaction system to nitrogen three times, react and stir at 90°C for 44 hours, and then cool. Cool to room temperature.
  • Step 1 Dissolve 4-chloro-3-oxobutyric acid ethyl ester (int-7a, 10 g, 60.75 mmol) in AcOH (48 mL) solution, slowly add NaNO 2 (5.2 g, 75.94 mmol) in H 2 O (40 mL) solution at 0°C, the reaction system turns red, and then warms to room temperature and stirs overnight. Add 100 mL H 2 O to the reaction solution, extract with 100 mL of ethyl acetate 3 times, wash with saturated brine (100 mL), dry with anhydrous sodium sulfate, and concentrate under reduced pressure.
  • Step 2 Add 4-chloro-2-(hydroxyimino)-3-oxobutyric acid ethyl ester (int-7b, 12g, 61.98mmol) and urea (29.8g, 495.87mmol) to DMF (50mL) solution in sequence, heat to 100°C and stir for 20min.
  • Step 1 4-methoxycyclohexan-1-one (int-8a, 1 g, 7.8 mmol), 1,1,1-trifluoro-N-phenyl-N-(trifluoro- To dry THF (20 mL) was added 2-fluoromethyl)sulfonyl)methanesulfonamide (2.8 g, 7.8 mmol), the temperature was lowered to -78 °C, and then a THF solution of LiHMDS (1 N, 7.8 mL) was added dropwise. The mixture was stirred at this temperature for 2 hours, then slowly returned to room temperature and stirred overnight. The mixture was quenched with saturated aqueous ammonium chloride solution and extracted with EA (20 mL x 2).
  • Step 1 In a 100 mL single-necked bottle, add tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-1, 1000 mg, 2.34 mmol), 2-iodo-N-(2-methyl-4-trifluoromethylphenyl)acetamide (880 mg, 2.57 mmol), DIPEA (907 mg, 7.02 mmol) and solvent DMF (5 mL) in sequence and stir at room temperature overnight. The reaction was completed after LCMS monitoring.
  • Step 2 Add the reactants 4-(2-bromo-5-ethyl-4-(2-(2-methyl-4-(trifluoro tert-butyl 2-(2-(2-(4-(2-( ...
  • Step 3 In a 100 mL single-necked bottle, add tert-butyl 4-(2-(3,6-dihydropyran-4-yl)-5-ethyl-4-(2-(2-methyl-4-trifluoromethylphenylamino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (1-b, 500 mg, 0.77 mmol), dioxane hydrochloride solution (4 M, 5 mL) and solvent DCM (10 mL). Stir at room temperature for 2 hr until the solution becomes turbid.
  • Step 1 Add diethyl ethoxymethylenemalonate (3-a, 1.5 g, 6.94 mmol) and methylhydrazine sulfate (1.0 g, 6.94 mmol) to 10 mL of aqueous solution and stir at 100 °C for 6 hours. The reaction solution was cooled to room temperature, solids precipitated, and filtered to obtain crude 5-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (3-b, 350 mg, 2.06 mmol, 29.6% yield). MS Calcd: 170.17; MS Found: 171.11 ([M+H] + ).
  • Step 2 Dissolve 5-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (3-b, 250 mg, 1.47 mmol) in 6 mL of methanol/water (5:1) mixed solution, add potassium hydroxide (824 mg, 14.7 mmol), heat to room temperature and reflux with stirring overnight, TLC monitoring reaction completion, completely concentrate the reaction solution, slowly add 5N HCl solution to the residue until solid precipitates, and filter to obtain 5-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (3-c, 150 mg, 1.06 mmol, 71.9% yield). MS Calcd: 142.11; MS Found: 143.05 ([M+H] + ).
  • Step 3 5-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (3-c, 78 mg, 0.55 mmol) and 1-chloro- N,N,2-Trimethylpropyleneamine (80 mg, 0.60 mmol) was added to 5 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then 2-(2-(3,6-dihydropyran-4-yl)-5-ethyl-7-oxo-6-piperazine-1-yl-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)yl)-N-(2-methyl-4-trifluoromethylphenyl)acetamide (1-c, 150 mg, 0.27 mmol) and DIPEA (178 mg, 1.37 mmol) were added, and the temperature was continued to rise to 40 °C and stirred for 5 hours.
  • DIPEA 178 mg, 1.37 mmol
  • Step 1 5-Hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (3-c, 45 mg, 0.32 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (46 mg, 0.35 mmol) were added to 5 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 90 mg, 0.16 mmol) and DIPEA (102 mg, 0.8 mmol) were added, and the temperature was continued to rise to 40 °C and stirred for 5 hours.
  • DIPEA 102 mg, 0.8 mmol
  • Step 1 2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (int-3, 100 mg, 0.18 mmol), (R)-tert-butyl 2-methylpiperazine-1-carboxylate (180 mg, 0.9 mmol), silver tetrafluoroborate (35 mg, 0.18 mmol) were added to dry DMSO (2 mL), and then the temperature was raised to 120° C. under nitrogen protection, and stirred for 4 hours.
  • Step 2 (R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (14-a, 74 mg, 0.11 mmol) was added to DCM (5 mL), and then trifluoroacetic acid (1 mL) was added and stirred at room temperature for 4 hours.
  • Step 3 5-Hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (3-c, 17 mg, 0.11 mmol) was added to DCM (1 mL), followed by 1-chloro-N,N,2-trimethylpropyl-1-en-1-amine (16 mg, 0.11 mmol), stirred at room temperature for 2 hours, then added to a solution of (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 51 mg, 0.09 mmol) and DIPEA (70 mg, 0.54 mmol) in dichloromethane and stirred at room temperature overnight.
  • DIPEA 70 mg, 0.54 mmol
  • Step 1 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 60 mg, 0.10 mmol), DIPEA (13 mg, 0.10 mmol), 1-cyclopropyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (18-a, 60 mg, 0.3 mmol) were added to 2-methyl-2-butanol and the mixture was stirred for 2 h.
  • Step 1 Dissolve 3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (16-a, 100 mg, 0.59 mmol) in 6 mL of methanol/water (5:1) mixed solution, add potassium hydroxide (200 mg, 3.54 mmol), heat to reflux and stir overnight, monitor the reaction completion by TLC, completely concentrate the reaction solution, slowly add 5N HCl solution to the residue until solid precipitates, and filter to obtain the product 3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (16-b, 70 mg, 0.49 mmol, 83.8% yield). MS Calcd: 142.11; MS Found: 143.07 ([M+H] + ).
  • Step 2 3-Hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (16-b, 40 mg, 0.28 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (42 mg, 0.31 mmol) were added to 5 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1, 2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-280 mg, 0.14 mmol) and DIPEA (90 mg, 0.71 mmol) were added, and the temperature was continued to rise to 40 °C and stirred for 5 h.
  • DIPEA 90 mg, 0.71 mmol
  • Step 2 Add 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 50 mg, 0.24 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2 45 mg, 0.06 mmol) to 4 mL of 2-methyl-2-butanol solution, place in a microwave reactor, heat to 180 ° C and stir for 30 min.
  • Step 1 Add diethyl ethoxymethylenemalonate (17-a, 2.0 g, 9.22 mmol), cyclopropylhydrazine hydrochloride (1.0 g, 9.22 mmol) and potassium carbonate (2.55 g, 18.43 mmol) to 10 mL of aqueous solution, respectively, and stir at 100 ° C for 6 hours.
  • Step 2 1-cyclopropyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (18-a, 117 mg, 0.6 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2 45 mg, 0.06 mmol) were added to 4 mL 2-methyl-2-butanol solution, placed in a microwave reactor and heated to 180°C and stirred for 30 min.
  • Step 1 Add tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-1, 2.2 g, 5.15 mmol), N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (2.0 g, 5.67 mmol) and DIPEA (2.0 g, 15.45 mmol) into 60 mL of dioxane solution respectively and stir at 80 °C for 4 hours. The reaction was completed by TLC monitoring, and the reaction solution was concentrated under reduced pressure.
  • Step 2 tert-Butyl 4-(2-bromo-5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (19-a, 1.44 g, 2.18 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (687 mg, 3.27 mmol), Pd(dppf)Cl 2 ⁇ CH 2 Cl 2 (357 mg, 0.44 mmol) and potassium phosphate (1.39 g, 6.54 mmol) were added successively into 20 mL of a mixed solution of dioxane/water (4:1), the mixture was evacuated and replaced with nitrogen three times, the temperature was raised to 80°C and stirred for 2 h.
  • Step 3 Add tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (19-b, 1.4 g, 2.11 mmol) to 20 mL of hydrochloric acid-dioxane solution and stir at room temperature overnight.
  • Step 4 Add 1-cyclopropyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (18-a, 80 mg, 0.41 mmol) and 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (19-c, 50 mg, 0.09 mmol) to 4 mL of 2-methyl-2-butanol solution, place in a microwave reactor, heat to 180 ° C and stir for 30 min.
  • Step 1 tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2a, 550 mg, 0.83 mmol) and 2-(4-methoxycyclohex-1-en-1-yl)-4,4,5-tetramethyl-1,3,2-dioxaborolato Alkane (237 mg, 1 mmol) was dissolved in 1,4-dioxane (8 mL) and H 2 O (4 mL), and Pd(dppf)Cl 2 (60 mg, 0.09 mmol) and potassium phosphate (570 mg, 2.49 mmol) were added in sequence.
  • 1,4-dioxane 8 m
  • Step 2 Dissolve tert-butyl 4-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (20-a, 370 mg, 0.53 mmol) in 8 mL of dichloromethane solution, add 2 mL of trifluoroacetic acid and stir at room temperature for 2 hours.
  • Step 3 Place N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (20-b, 50 mg, 0.08 mmol) and 5-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (15 mg, 0.10 mmol) in a 25 mL eggplant-shaped bottle, add 5 mL of DCM to dissolve, and then add PyBOP (43 mg, 0.08 mmol) and DIPEA (0.04 mL, 0.22 mmol) in sequence, and react at room temperature for 3 hours.
  • Step 1 Add diethyl ethoxymethylenemalonate (17-a, 1.95 g, 9.04 mmol), isopropylhydrazine hydrochloride (1.0 g, 9.04 mmol) and potassium carbonate (2.05 g, 18.08 mmol) to 10 mL of aqueous solution, respectively, and stir at 100°C for 6 hours.
  • Step 2 Place the compound 5-hydroxy-1-isopropyl-1H-pyrazole-4-carboxylic acid ethyl ester (21-a, 83 mg, 0.42 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (20-b, 50 mg, 0.08 mmol) in a 25 mL eggplant shaped bottle, and add 2 mL of 2-methyl-2-butanol.
  • Step 1 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 60 mg, 0.10 mmol), DIPEA (13 mg, 0.10 mmol), 5-hydroxy-1-isopropyl-1H-pyrazole-4-carboxylic acid ethyl ester (21-a, 102 mg, 0.5 mmol) were added to 2-methyl-2-butanol (1 mL).
  • Step 1 Weigh 3-hydroxy-2-methylquinoline-4-carboxylic acid (26-a, 19 mg, 0.09 mmol), PyBOP (46.8 mg, 0.09 mmol), N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 60 mg, 0.09 mmol) into a 25 mL single-necked bottle, add DCM (2 mL) and DIPEA (0.04 mL, 0.27 mmol), and stir at room temperature overnight after addition.
  • DCM 2- mL
  • DIPEA 0.27 mmol
  • Step 1 Add 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 80 mg, 0.38 mmol) and 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (19-c, 50 mg, 0.09 mmol) to 4 mL of 2-methyl-2-butanol solution, place in a microwave reactor, heat to 180 ° C and stir for 30 min.
  • Step 1 5-Hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (3-c, 40 mg, 0.28 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (42 mg, 0.31 mmol) were added to 5 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (19-c, 80 mg, 0.14 mmol) and DIPEA (90 mg, 0.71 mmol) were added, and the temperature was continued to rise to 40 ° C and stirred for 5 hours.
  • DIPEA 90 mg, 0.71 mmol
  • Step 1 Dissolve 3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (16-a, 500 mg, 2.94 mmol) in 10 mL of dichloromethane solution, slowly add sulfonyl chloride (0.47 g, 3.53 mmol) at 0°C, slowly warm to room temperature and stir overnight. Add water to the reaction solution to terminate the reaction, extract with dichloromethane, concentrate under reduced pressure, and purify the residue by prep-HPLC to obtain 5-chloro-3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (29-a, 120 mg, 0.59 mmol, 20% yield). MS Calcd: 204.03; MS Found: 205.10 ([M+H] + ).
  • Step 2 Dissolve 5-chloro-3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (29-a, 120 mg, 0.59 mmol) in 4 mL of methanol/water (5:1) mixed solution, add potassium hydroxide (200 mg, 3.52 mmol), heat to reflux and stir overnight, completely concentrate the reaction solution, slowly add 5N HCl solution to the residue until solid precipitates, and filter to obtain 5-chloro-3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (29-b, 90 mg, 0.51 mmol, 86.9% yield). MS Calcd: 176.00; MS Found: 177.01 ([M+H] + ).
  • Step 3 5-Chloro-3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (29-b, 44 mg, 0.25 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (36 mg, 0.27 mmol) were added to 5 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 70 mg, 0.12 mmol) and DIPEA (80 mg, 0.62 mmol) were added, and the temperature was continued to rise to 40 °C and stirred for 5 hours.
  • DIPEA 80 mg, 0.62 mmol
  • Step 1 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (19-c, 50 mg, 0.09 mmol) and 5-hydroxy-1-isopropyl-1H-pyrazole-4-carboxylic acid ethyl ester (21-a, 87.9 mg, 0.44 mmol) were added to 4 mL of 2-methyl-2-butanol solution, placed in a microwave reactor, heated to 180 ° C and stirred for 30 min.
  • Step 1 Add 5-hydroxy-1-isopropyl-1H-pyrazole-4-carboxylic acid ethyl ester (21-a, 90 mg, 0.44 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 50 mg, 0.09 mmol) to 4 mL of 2-methyl-2-butanol solution, place in a microwave reactor, heat to 180 ° C and stir for 30 min.
  • Step 1 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 55 mg, 0.10 mmol), DIPEA (13 mg, 0.10 mmol), 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 63 mg, 0.3 mmol) were added to 2-methyl-2-butanol and the mixture was stirred for 2 h.
  • Step 1 5-Chloro-3-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (29-b, 30 mg, 0.17 mmol) was added to DCM (1 mL), followed by 1-chloro-N,N,2-trimethylpropyl-1-en-1-amine (23 mg, 0.17 mmol), stirred at room temperature for 2 hours, and then added to (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 40 mg, 0.07 mmol) and DIPEA (54 mg, 0.42 mmol).
  • Step 1 Place 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 88 mg, 0.42 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (20-b, 50 mg, 0.08 mmol) in a 25 mL eggplant-shaped bottle, and add 2 mL of 2-methyl-2-butanol.
  • Step 1 Place tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-1, 200 mg, 0.47 mmol) in a 100 mL eggplant-shaped bottle, add 10 mL of 1,4-dioxane solution. Then add N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (int-4, 185.9 mg, 0.51 mmol) and DIPEA (0.23 mL, 1.4 mmol) in sequence.
  • Step 2 tert-Butyl 4-(2-bromo-5-ethyl-4-(2-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (35-a, 180 mg, 0.27 mmol) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (85.9 mg, 0.41 mmol) were dissolved in 1,4-dioxane (6 mL) and H 2 O (3 mL), and Pd(dppf)Cl 2 was added in sequence.
  • Step 3 Dissolve tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (35-b, 153 mg, 0.23 mmol) in 8 mL of dichloromethane solution, add 2 mL of trifluoroacetic acid and stir at room temperature for 2 hours.
  • Step 4 Compound 3-hydroxy-2-methylquinoline-4-carboxylic acid (27.0 mg, 0.13 mmol) and HATU (102.8 mg, 0.27 mmol) were placed in a 25 mL eggplant-shaped bottle, 5 mL of DCM was added, and then DIPEA (0.04 mL, 0.27 mmol) was added, and stirred for 2 minutes after the addition was complete.
  • Step 1 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 88 mg, 0.42 mmol) and 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidine -4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (35-c, 50 mg, 0.08 mmol) was placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added, followed by stirring at 180° C.
  • Step 1 Dissolve (2,2,2-trifluoroethyl)hydrazine hydrochloride (37-a, 4000 mg, 35.06 mmol) in 40 mL of water, then slowly add diethyl 2-(ethoxymethylene)malonate (7579.3 mg, 35.06 mmol) and K 2 CO 3 (9675 mg, 17.53 mmol) at 0° C., then heat to 100° C. and stir for 5 h. TLC indicates that the reaction of the starting material is complete. The reaction solution was cooled to room temperature, and some of the solvent was dried by rotary evaporation.
  • Step 2 5-hydroxy-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (37-b, 100 mg, 0.44 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (20-b, 50 mg, 0.08 mmol) were placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added.
  • Step 1 Weigh 5-chloro-2-methyl-4-(trifluoromethyl)aniline (38-a, 6.0 g, 28.63 mmol) into a 50 mL single-mouth bottle, add DCM (30 mL), then add triethylamine (7.94 mL, 57.25 mmol), add chloroacetyl chloride (3.40 mL, 34.35 mmol) in DCM solution (10 mL) dropwise under ice bath, then naturally return to room temperature to react overnight.
  • Step 2 Add 2-chloro-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (38-b, 3.7 g, 12.93 mmol) and solvent acetone (15 mL) to a 100 mL eggplant-shaped bottle. Then add potassium iodide (5.4 g, 32.33 mmol) and stir at 50°C for 3 hours.
  • Step 3 Add N-[5-chloro-2-methyl-4-(trifluoromethyl)phenyl]-2-iodoacetamide (38-c, 1.1 g, 2.81 mmol), tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-1, 1.0 g, 2.34 mmol) and solvent 1,4-dioxane (15 mL) to a 100 mL eggplant-shaped bottle. Then add DIPEA (0.9 g, 7.02 mmol) and stir at 90°C for 2 hours.
  • DIPEA 0.9 g, 7.02 mmol
  • Step 4 Add 4-(2-bromo-4-(2-(2-chloro-5-methyl-3-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1- Tert-butyl formate (38-d, 940 mg, 1.39 mmol) and solvent 1,4-dioxane (10 mL), H 2 O (5 mL) were added.
  • Step 5 tert-Butyl 4-(4-(2-(2-chloro-5-methyl-3-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (38-e, 800 mg, 1.18 mmol) and solvent DCM (5 mL) were added to a 100 mL eggplant-shaped bottle. TFA (0.5 mL) was then added and stirred at room temperature overnight.
  • solvent DCM 5 mL
  • Step 1 Add 5-hydroxy-1-isopropyl-1H-pyrazole-4-carboxylic acid ethyl ester (21-a, 85 mg, 0.43 mmol) and N-(2-chloro-5-methyl-3-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (38-f, 50 mg, 0.09 mmol) to 4 mL of 2-methyl-2-butanol solution, place in a microwave reactor, heat to 180 ° C and stir for 30 min.
  • Step 2 5-Hydroxy-1-methyl-1H-pyrazole-3-carboxylic acid (40-b, 34 mg, 0.24 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (36 mg, 0.27 mmol) were added to 5 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 70 mg, 0.12 mmol) and DIPEA (80 mg, 0.62 mmol) were added, and the temperature was continued to rise to 40 °C and stirred for 5 hours.
  • DIPEA 80 mg, 0.62 mmol
  • Step 1 Weigh 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 65 mg, 0.30 mmol), N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 40 mg, 0.06 mmol) into a 25 mL single-necked bottle, add 2-methyl-1-butanol (2 mL) and DIPEA (0.01 mL, 0.06 mmol), and after addition, heat to 180 degrees Celsius and stir for 30 minutes.
  • 2-methyl-1-butanol 2 mL
  • DIPEA 0.01 mL, 0.06 mmol
  • Step 1 Add diethyl ethoxymethylenemalonate (17-a, 1.74 g, 8.03 mmol), tert-butylhydrazine hydrochloride (1.0 g, 8.03 mmol) and potassium carbonate (2.21 g, 16.05 mmol) to 10 mL of aqueous solution, respectively, and stir at 100°C for 6 hours. Adjust the pH to 2 with 5N HCl, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate under reduced pressure.
  • Step 2 1-(tert-butyl)-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (42-a, 94 mg, 0.44 mmol) and 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a] Pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (19-c, 50 mg, 0.09 mmol) was added to 2 mL of 2-methyl-2-butanol solution, placed in a microwave reactor and heated to 180°C and stirred for 30 min.
  • Step 2 Dissolve N'-(1,1,1-trifluoropropane-2-ylidene)benzohydrazide (43-c, 3.0 g, 13.3 mmol) in 40 mL of tetrahydrofuran solution, slowly add borane tetrahydrofuran solution (26.06 mL, 26.06 mmol) at 0 ° C, slowly warm to room temperature and stir overnight.
  • Step 3 Dissolve N'-(1,1,1-trifluoropropane-2-yl)benzohydrazide (43-d, 1.0 g, 4.31 mmol) in 5 mL of methanol solution, then add 8 mL of concentrated hydrochloric acid, heat to 100 °C and stir overnight. TLC monitoring of raw materials The reaction mixture disappeared, and the reaction solution was completely concentrated to obtain the crude product (1,1,1-trifluoropropan-2-yl)hydrazine hydrochloride (43-e, 1.5 g, 9.2 mmol).
  • Step 5 Add 5-hydroxy-1-(1,1,1-trifluoropropan-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (43-f, 140 mg, 0.54 mmol) and 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (19-c, 50 mg, 0.09 mmol) to 4 mL of 2-methyl-2-butanol solution, place in a microwave reactor, heat to 180 ° C and stir for 30 min.
  • Step 1 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 50 mg, 0.09 mmol), DIPEA (13 mg, 0.10 mmol), 5-hydroxy-1-(1, 1,1-trifluoropropan-2-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (43-f, 126 mg, 0.5 mmol) was added to 2-methyl-2-butanol (1 mL), the oil bath was heated to 180°C, stirred for 30 minutes, the reaction system was dissolved in methanol, and directly purified by prep-HPLC to obtain the title compound: N-(2-chloro-4-(
  • Step 1 Dissolve 2-(ethoxycarbonyl)-3-oxobutyric acid ethyl ester (45-a, 1 g, 4.95 mmol) and isopropylhydrazine hydrochloride (45-b, 0.4 g, 4.95 mmol) in EtOH (20 mL), add HCl (1 mL, 4.95 mmol), and react at 85°C for 2 hours.
  • Step 2 Add 5-hydroxy-1-isopropyl-3-methyl-1H-pyrazole-4-carboxylic acid ethyl ester (45-c, 104 mg, 0.45 mmol) and 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (19-c, 50 mg, 0.09 mmol) to 4 mL of 2-methyl-2-butanol solution, heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 Dissolve ethyl 4-chloroacetoacetate (47-a, 2.2 g, 13.37 mmol) in 10 mL of acetic acid, place at 0°C, slowly add an aqueous solution (10 mL) of sodium nitrite (1.2 g, 16.71 mmol), slowly warm to room temperature and stir overnight. Add water to the reaction solution, extract with ethyl acetate, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate under reduced pressure.
  • Step 2 Add 4-chloro-2-(hydroxyimino)-3-oxobutanoic acid ethyl ester (47-b, 500 mg, 2.58 mmol) to 3 mL of DMF solution, then add urea (1.24 g, 20.66 mmol) and stir at 100°C for 3 h. Add water to the reaction solution, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate under reduced pressure.
  • Step 3 Add 4-hydroxyisoxazole-3-carboxylic acid ethyl ester (47-c, 70 mg, 0.44 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 50 mg, 0.09 mmol) to 4 mL of 2-methyl-2-butanol solution, place in an oil bath, heat to 180°C and stir for 30 min.
  • 4-hydroxyisoxazole-3-carboxylic acid ethyl ester 47-c, 70 mg, 0.44 mmol
  • Step 1 Add N-(2-chloro-5-methyl-3-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (38-f, 50 mg, 0.09 mmol) and solvent 2-methyl-2-butanol (2 mL) to a 50 mL eggplant-shaped bottle.
  • Step 1 Add compound N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 100 mg, 0.18 mmol) and solvent 2-methyl-2-butanol (1.5 mL) to a 50 mL eggplant-shaped bottle.
  • Step 1 Add compound 2-chloro-6-(trifluoromethyl)pyridin-3-amine (51-a, 4.8 g, 24.42 mmol) and solvent dichloromethane (30 mL) to a 250 mL eggplant-shaped bottle, then add triethylamine (4.9 g, 48.83 mmol) and chloroacetyl chloride (2.8 g, 24.42 mmol) and stir at room temperature overnight. Add 150 mL of dichloromethane to dilute, wash the organic phase with water (100 mL*2), and then dry it over anhydrous sodium sulfate.
  • Step 3 Add compound N-[2-chloro-6-(trifluoromethyl)pyridin-3-yl]-2-iodoacetamide (51-c, 457 mg, 1.25 mmol), tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-5, 450 mg, 1.05 mmol) and solvent 1,4-dioxane (15 mL) to a 100 mL eggplant-shaped bottle.
  • Step 4 tert-Butyl 4-(4-(2-(2-chloro-6-(trifluoromethyl)pyridin-3-yl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (51-d, 650 mg, 0.97 mmol) and solvent DCM (8 mL) were added to a 100 mL eggplant-shaped bottle. TFA (1 mL) was then added and stirred at room temperature overnight.
  • Step 5 Add N-[2-chloro-6-(trifluoromethyl)pyridin-3-yl]-2-[2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]acetamide (51-e, 70 mg, 0.12 mmol) and solvent 2-methyl-2-butanol (2 mL) to a 50 mL eggplant-shaped bottle.
  • Step 1 Add 2,5-dichloro-4-(trifluoromethyl)aniline (52-a, 800 mg, 3.48mmol) and solvent dichloromethane (10mL). Then add chloroacetyl chloride (589mg, 5.22mmol) and triethylamine (1056mg, 10.43mmol) and stir at room temperature for 2 hours. Add water (40mL) and extract with DCM (30mL).
  • Step 2 Add 2-chloro-N-(2,5-dichloro-4-(trifluoromethyl)phenyl)acetamide (52-b, 1000 mg, 3.26 mmol) and solvent acetone (20 mL) to a 25 mL eggplant-shaped bottle. Then add potassium iodide (1624.8 mg, 9.79 mmol) and heat to 50°C and stir for 2 hours.
  • Step 3 Add the compound 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester (int-5, 200 mg, 0.46 mmol) and solvent 1,4-dioxane (10 mL) to a 25 mL eggplant-shaped bottle.
  • Step 4 4-(4-(2-((2,5-dichloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester (52-d, 100 mg, 0.14 mmol) and solvent dichloromethane (50 mL) were added to a 25 mL eggplant-shaped bottle.
  • Step 5 Add N-(2,5-dichloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (52-e, 35 mg, 0.06 mmol) and solvent tert-amyl alcohol (2 mL) to a 25 mL eggplant-shaped bottle.
  • Step 1 5-hydroxy-1-(2,2,2-trifluoroethyl)-1H-pyrazole-4-carboxylic acid ethyl ester (37-b, 105 mg, 0.44 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 50 mg, 0.08 mmol) were placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added.
  • Step 1 Add 2-bromo-5-ethyl-4,7-dihydro[1,2,4]triazolo[1,5-a]pyrimidin-7-one (int-3a, 700 mg, 2.88 mmol) and N-[5-chloro-2-methyl-4-(trifluoromethyl)phenyl]-2-iodoacetamide (38-c, 1087.0 mg, 2.88 mmol) to a 100 mL round-bottom flask, add dioxane (20 mL) and DIPEA (1.43 mL, 8.64 mmol) in sequence, and stir at 80 °C for 3 hours under nitrogen protection.
  • Step 2 2-(2-Bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (54-a, 600 mg, 1.22 mmol) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (383.8 mg, 1.83 mmol) were dissolved in 1,4-dioxane (8 mL) and H 2 O (4 mL), and Pd(dppf)Cl 2 (89 mg, 0.09 mmol) and potassium phosphate (860 mg, 3.65 mmol) were added in sequence.
  • Step 3 Place N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (54-b, 450 mg, 0.91 mmol) and N-bromosuccinimide (192.7 mg, 1.09 mmol) in a 100 mL eggplant flask, add 10 mL of acetonitrile, and then raise the temperature to 60 °C for reaction for 1 hour. The reaction solution was poured into 20 mL of water and extracted with EA (3 ⁇ 20 mL).
  • Step 4 2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (54-c, 250 mg, 0.43mmol) and compound 2-methylpropan-2-yl (2R)-2-methylpiperazine-1-carboxylate (871mg, 4.35mmol), AgBF4 (101.3mg, 0.52mmol) were added to dry DMSO (3mL), nitrogen was replaced for 1 minute, and then the temperature was raised to 120°C for reaction for 18h.
  • Step 5 Dissolve (R)-4-(4-(2-((5-chloro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (54-d, 127 mg, 0.18 mmol) in 8 mL of dichloromethane solution, add 2 mL of trifluoroacetic acid and stir at room temperature for 2 hours.
  • Step 6 (R)-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (54-e, 50 mg, 0.08 mmol) and 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 88 mg, 0.42 mmol) were placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added.
  • Step 1 (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 63 mg, 0.10 mmol), DIPEA (15 mg, 0.10 mmol), 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 112 mg, 0.5 mmol) were added to 2-methyl-2-butanol (1 mL).
  • Step 1 Add 2-bromo-5-ethyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (int-3a, 4 g, 16.5 mmol), N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (int-4, 6 g, 16.5 mmol), DIPEA (6.4 g, 49.5 mmol) to DMF (50 mL), raise the temperature to 80°C, stir for 2 hours, After cooling to room temperature, the reaction system was poured into water. Solids precipitated and filtered. The filter cake was washed with water (50 mL x 2).
  • Step 2 2-(2-bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (56-a, 5.9 g, 12.4 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborane (2.6 g, 12.4 mmol), potassium phosphate (7.9 g, 37.2 mmol), 1,1-bis(diphenylphosphino)diferronichloridopalladium (906 mg, 1.24 mmol) were added.
  • Step 3 Add 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (56-b, 4.8 g, 10.0 mmol) to DMF (50 mL), then add NBS (1.78 g, 10.0 mmol), raise the temperature to 60°C and stir for 3 hours. The reaction system was then poured into water, and solids precipitated.
  • Step 4 2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (56-c, 1 g, 1.8 mmol), (R)-tert-butyl 2-methylpiperazine-1-carboxylate (3.6 g, 18 mmol), and silver tetrafluoroborate (351 mg, 1.8 mmol) were added to dry DMSO (10 mL), and then the temperature was raised to 120°C under nitrogen protection, stirred for 4 hours, cooled to room temperature, and saturated Sodium bicarbonate aqueous solution, then extracted with ethyl acetate (10 mL x 3), the organic layer was dried over anhydrous sodium sulfate, filtered, the filtrate
  • Step 5 (R)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(-2-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (56-d, 810 mg, 1.20 mmol) was added to DCM (10 mL), and then trifluoroacetic acid (3 mL) was added, stirred at room temperature for 4 hours, concentrated under reduced pressure, saturated aqueous sodium bicarbonate solution was added to the residue, and then extracted with DCM (10 mL x 3), the organic layer was dried over anhydrous sodium sulfate, filtered, and the filtrate was purified by centrifugation.
  • Step 6 (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (56-e, 60 mg, 0.10 mmol), DIEA (15 mg, 0.10 mmol), 1-cyclopropyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (18-a, 100 mg, 0.5 mmol) were added to 2-methyl-2-butanol ( 1mL), the oil bath was heated to 180°C, stirred for 30 minutes, and purified by prep-HPLC to obtain the title compound: (R)-2-(6-(4-(1-cyclopropyl-5-hydroxy-1H-pyrazole-4-yl
  • Step 1 tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2a, 100 mg, 0.15 mmol) and solvent DMSO (2 mL) were added to a 25 mL eggplant-shaped bottle. N-methylpiperazine (45.1 mg, 0.45 mmol) and potassium acetate (44.1 mg, 0.45 mmol) were then added, and the mixture was heated to 120° C.
  • Step 2 Add 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxo- 1-(4-methylpiperazine-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-5-ethyl 1-2-(4-methylpiperazine-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-1-piperazine-1-carboxylic acid tert-butyl ester (57-a, 45 mg, 0.07 mmol, 44% yield) and solvent dichloromethane (5 mL).
  • Step 3 Add N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(4-methylpiperazin-1-yl)-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (57-b, 40 mg, 0.07 mmol) and solvent tert-amyl alcohol (2 mL) to a 25 mL eggplant-shaped bottle.
  • Step 1 Add N-[2-chloro-6-(trifluoromethyl)pyridin-3-yl]-2-[2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]acetamide (51-e, 70 mg, 0.12 mmol) and solvent 2-methyl-2-butanol (2 mL) to a 50 mL eggplant-shaped bottle.
  • Step 1 Weigh 4-hydroxy-8-methoxyquinoline-3-carboxylic acid (59-a, 23 mg, 0.09 mmol), N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 40 mg, 0.06 mmol) into a 25 mL single-necked bottle, add 2-methyl-2-butanol (2 mL) and DIPEA (0.01 mL, 0.06 mmol), and after addition, heat to 180 °C and react for 2 hours.
  • 2-methyl-2-butanol 2 mL
  • DIPEA 0.01 mL, 0.06 mmol
  • Step 1 4-Hydroxyisoxazole-3-carboxylic acid (int-7, 26 mg, 0.20 mmol) was added to DCM (1 mL), followed by 1-chloro-N,N,2-trimethylpropyl-1-en-1-amine (29 mg, 0.20 mmol), stirred at room temperature for 2 hours, then added to a solution of (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 60 mg, 0.10 mmol) and DIPEA (64 mg, 0.50 mmol) in dichloromethane, and stirred at room temperature for 2 hours.
  • DIPEA 64 mg, 0.50 mmol
  • Step 1 4-Hydroxyisoxazole-3-carboxylic acid (int-7, 26 mg, 0.20 mmol) was added to DCM (1 mL), followed by 1-chloro-N,N,2-trimethylpropyl-1-en-1-amine (29 mg, 0.20 mmol), stirred at room temperature for 2 hours, and then added to (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (56-e, 60 mg, 0.10 mmol) and DIPEA (64 mg, 0.5 0mmol) in dichloromethane solution and stirred at room temperature overnight, then water (3mL) was added, extracted with DCM (3
  • Step 1 Add tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2a, 200 mg, 0.30 mmol) and solvent DMSO (2 mL) to a 25 mL eggplant-shaped bottle. Then add morpholine (78.8 mg, 0.91 mmol) and potassium acetate (88.8 mg, 0.91 mmol), and then heat to 120°C and stir overnight.
  • morpholine 78.8 mg, 0.91 mmol
  • potassium acetate 88.8 mg, 0.91 mmol
  • Step 2 Add 4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyll)-5-ethyl-2-morpholine-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yll))piperazine-1-carboxylic acid tert-butyl ester (62-a, 55 mg, 0.08 mmol) and solvent dichloromethane (5 mL) to a 25 mL eggplant-shaped bottle. Then add hydrogen chloride-1,4-dioxane solution (2 mL, 4 mol/L) and stir at room temperature for 1 hour.
  • Step 3 Add N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-morpholino-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (62-b, 40 mg, 0.08 mmol) and solvent tert-amyl alcohol (2 mL) to a 25 mL eggplant-shaped bottle.
  • Step 1 Add 4-hydroxyisoxazole-3-carboxylic acid (int-7, 12.9 mg, 0.10 mmol) and solvent DCM (3 mL) to a 10 mL eggplant-shaped bottle. Then add 1-chloro-N,N,2-trimethylpropenamine (10 mg, 0.08 mmol), stir at room temperature for 1 hour, add N-(2,5-dichloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (52-e, 30 mg, 0.05 mmol) and DIPEA (19.4 mg, 0.15 mmol), continue stirring at room temperature for 2 hours, and add water (20 mL) , extracted with DCM (20 mL), washed with saturated brine (20 m
  • Step 1 Add compound N-[2-chloro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (8.2 g, 22.62 mmol), 2-bromo-5-ethyl-4,7-dihydro[1,2,4]triazolo[1,5-a]pyrimidin-7-one (int-3a, 5.0 g, 20.57 mmol) and solvent 1,4-dioxane (120 mL) to a 250 mL eggplant-shaped bottle. Then add DIPEA (8.0 g, 61.70 mmol) and stir at 80°C for 4 hours.
  • DIPEA 8.0 g, 61.70 mmol
  • Step 2 2-(2-bromo-5-ethyl-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-4-yl)-N-[2-chloro-4-(trifluoromethyl)phenyl]acetamide (64-a, 800 mg, 1.67 mmol), 2-(4-methoxycyclohex-1-enyl)-4,4,5-5-tetramethyl-1,3,2-dioxaborane (400 mg, 1.67 mmol) and solvent 1,4-dioxane (6 mL), H 2 O (2 mL) were added to a 100 mL eggplant-shaped bottle.
  • Step 3 Add N-[2-chloro-4-(trifluoromethyl)phenyl]-2-[5-ethyl-2-(4-methoxycyclohex-1-enyl)-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]acetamide (64-b, 650 mg, 1.27 mmol) and acetonitrile (10 mL) to a 100 mL eggplant-shaped bottle. Then add NBS (250 mg, 1.40 mmol) and stir at 60°C for 1 hour. The solvent was dried by vortexing, 20 mL of water was added, and the mixture was extracted with ethyl acetate (20 mL ⁇ 3).
  • Step 4 Add 2-[6-bromo-5-ethyl-2-(4-methoxycyclohex-1-enyl)-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]-N-[2-chloro-4-(trifluoromethyl)phenyl]acetamide (64-c, 500 mg, 0.85mmol), 2-methylpropan-2-yl (2R)-2-methylpiperazine-1-carboxylate (1700mg, 8.49mmol) and solvent DMSO (10mL) were added, and then silver tetrafluoroborate (198mg, 1.02mmol) was added thereto, and stirred at 120°C for 24 hours.
  • Step 5 2-methylpropan-2-yl (2R)-4-[4-(2- ⁇ [2-chloro-4-(trifluoromethyl)phenyl]amino ⁇ -2-oxoethyl)-5-ethyl-2-(4-methoxycyclohex-1-enyl)-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-6-yl]-2-methylpiperazine-1-carboxylate (64-d, 340 mg, 0.48 mmol) and solvent DCM (3 mL) were added to a 100 mL eggplant-shaped bottle. TFA (0.5 mL) was then added and stirred at room temperature for 3 hours.
  • Step 6 N-[2-chloro-4-(trifluoromethyl)phenyl]-2-[5-ethyl-2-(4-methoxycyclohex-1-enyl)-6-[(3R)-3-methylpiperazin-1-yl]-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]acetamide (64-e, 60 mg, 0.10 mmol) and solvent 2-methyl-2-butanol (1.5 mL) were added to a 50 mL eggplant-shaped bottle.
  • 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 83 mg, 0.39 mmol) and DIPEA (39 mg, 0.30 mmol) were added and stirred at 180° C. for 30 minutes.
  • Step 1 Add compound 4-hydroxyisoxazole-3-carboxylic acid (int-7, 31 mg, 0.24 mmol) and solvent DCM (1 mL) to a 50 mL eggplant-shaped bottle. Then add (1-chloro-2-methylprop-1-enyl)dimethylamine (33 mg, 0.25 mmol) and stir at room temperature for 3 hours.
  • Step 1 Add 5-hydroxy-1-methyl-1H-pyrazole-4-carboxylic acid (3-c, 12 mg, 0.08 mmol) and solvent DCM (1 mL) to a 50 mL eggplant-shaped bottle. Then add (1-chloro-2-methylprop-1-enyl)dimethylamine (12 mg, 0.09 mmol) and stir at room temperature for 3 hours.
  • Step 1 4-Hydroxyisoxazole-3-carboxylic acid (int-7, 26.1 mg, 0.20 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (29.7 mg, 0.22 mmol) were added to 3 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (20-b, 60 mg, 0.10 mmol) and DIEA (0.05 mL, 0.3 mmol) were added, and the temperature was continued to rise to 40 °C and stirred for 5 hours.
  • DIEA 0.05 mL, 0.3 mmol
  • Step 1 Place 2-iodo-N-(2,5-dichloro-4-(trifluoromethyl)phenyl)acetamide (52-c, 420 mg, 1.06 mmol) in a 100 mL eggplant-shaped bottle, add 10 mL of 1,4-dioxane solution. Then add 2-bromo-5-ethyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (int-3a, 250 mg, 1.03 mmol) and DIPEA (0.5 mL, 3.09 mmol) in sequence. Stir at 75°C for 5 hours.
  • Step 2 2-(2-Bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2,5-dichloro-4-(trifluoromethyl)phenyl)acetamide (68-a, 300 mg, 0.58 mmol) and 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (184.3 mg, 0.88 mmol) were dissolved in 1,4-dioxane (8 mL) and H 2 O (4 mL), and Pd(dppf)Cl 2 (42.7 mg, 0.04 mmol) and potassium phosphate (403 mg, 1.75 mmol) were added in sequence.
  • Step 3 Place N-(2,5-dichloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (68-b, 220 mg, 0.33 mmol) and N-bromosuccinimide (69.9 mg, 0.40 mmol) in a 100 mL eggplant-shaped bottle, add 10 mL of acetonitrile, and then raise the temperature to 60°C for reaction for 1 hour. The reaction solution was poured into 20 mL of water and extracted with EA (3 ⁇ 20 mL).
  • Step 4 2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2,5-dichloro-4-(trifluoromethyl)phenyl)acetamide (68-c, 50 mg, 0.08 mmol) and (R)-1-N-Boc-2-methylpiperazine (168 mg, 0.84 mmol), AgBF 4 (16.4 mg, 0.08 mmol) were added to dry DMSO (1 mL), nitrogen was replaced for 1 minute, and then the temperature was raised to 120° C. for reaction for 18 h.
  • Step 5 Dissolve (R)-4-(4-(2-((2,5-dichloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (68-d, 25 mg, 0.03 mmol) in 4 mL of dichloromethane solution, add 1 mL of trifluoroacetic acid and stir at room temperature for 2 hours.
  • Step 6 4-Hydroxyisoxazole-3-carboxylic acid (int-7, 12.6 mg, 0.10 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (13.0 mg, 0.10 mmol) were added to 3 mL of dichloromethane solution, stirred at room temperature for 3 hours, and then (R)-N-(2,5-dichloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (68-e, 20 mg, 0.03 mmol) and DIPEA (0.02 mL, 0.1 mmol) were added, and the temperature was continued to rise to 40 °C and stirred for 5 hours.
  • DIPEA
  • Step 1 1-cyclopropyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (18-a, 82 mg, 0.42 mmol) and (R)-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (54-e, 50 mg, 0.08 mmol) were placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added.
  • Step 1 4-Hydroxyisoxazole-3-carboxylic acid (int-7, 32.6 mg, 0.25 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (33.7 mg, 0.25 mmol) were added to 3 mL of dichloromethane solution respectively, stirred at room temperature for 3 hours, and then (R)-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (54-e, 50 mg, 0.08 mmol) and DIPEA (0.04 mL, 0.2 mmol) were added, and the temperature was continued to rise to 40°C and stirred for 5 hours.
  • DIPEA 0.0
  • Step 1 Weigh 4-hydroxyisoxazole-3-carboxylic acid (Int-7, 25 mg, 0.19 mmol), HATU (97.5 mg, 0.26 mmol) into a 25 mL single-necked bottle, add DCM (2 mL) and DIPEA (0.06 mL, 0.38 mmol), stir at room temperature for 5 minutes, then add 2-(2-(3,6-dihydropyran-4-yl)-5-ethyl-7-oxo-6-piperazine-1-yl-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)yl)-N-(2-methyl-4-trifluoromethylphenyl)acetamide (1-c, 70 mg, 0.13 mmol), react at room temperature for 1 hour.
  • Step 1 Add 2-bromo-5-ethyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (int-3a, 2.0 g, 8.23 mmol), N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (3.0 g, 8.23 mmol) and DIPEA (3.2 g, 24.68 mmol) into 60 mL of dioxane solution and stir at 80 °C for 4 hours.
  • Step 2 2-(2-Bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (72-a, 3.6 g, 7.56 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (1.58 g, 7.5 mmol), Pd(dppf)Cl 2 ⁇ CH 2 Cl 2 (1.07 g, 1.32 mmol) and potassium phosphate (4.77 g, 22.5 mmol) were added successively into 40 mL of a mixed solution of dioxane/water (4:1), the solution was evacuated and replaced with nitrogen three times, the temperature was raised to 80°C and stirred for 2 h.
  • Step 3 Dissolve 2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (72-b, 1.34 g, 2.8 mmol) in DMF (10 mL) solution, then slowly add NBS (0.55 g, 3.07 mmol), heat to 60 °C and stir for 3 h.
  • Step 4 2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (72-c, 1.3 g, 2.33 mmol), (R)-1-N-Boc-2-methylpiperazine (5.02 g, 25.08 mmol) and silver tetrafluoroborate (0.58 g, 3.01 mmol) were added to DMSO (10 mL) solution in turn, vacuumed, replaced with nitrogen 3 times, heated to 120°C and stirred overnight.
  • DMSO 10 mL
  • Step 5 (R)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(-2-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (72-d, 1.2 g, 1.77 mmol) was added to a hydrochloric acid-dioxane solution (20 mL) and stirred at room temperature overnight.
  • Step 6 Add 1-cyclopropyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (18-a, 102 mg, 0.52 mmol), DIPEA (40 mg, 0.31 mmol) and (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (72-e, 50 mg, 0.09 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 Add 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 105 mg, 0.5 mmol), DIPEA (40 mg, 0.31 mmol) and (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (72-e, 60 mg, 0.1 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 4-Hydroxyisoxazole-3-carboxylic acid (int-7, 40 mg, 0.31 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (44 mg, 0.33 mmol) were added to DCM solution (2 mL) respectively, stirred at room temperature for 3 hours, and then (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (72-e, 60 mg, 0.1 mmol) and DIPEA (80 mg, 0.62 mmol) were added, and the temperature was continued to rise to 40°C and stirred for 5 hours.
  • DIPEA 80 mg, 0.62 mmol
  • Step 1 Add 3-fluoro-4-(trifluoromethyl)aniline (75-a, 10 g, 55.83 mmol), DCM (50 mL) to a 250 mL eggplant-shaped bottle, and then add BF 3 OEt 2 (7.9 g, 55.83 mmol), NCS (8.2 g, 61.42 mmol), and stir at room temperature for 3 hours.
  • Step 2 Add 2-chloro-5-fluoro-4-(trifluoromethyl)aniline (75-b, 8.1 g, 37.92 mmol), DCM (50 mL) to a 250 mL eggplant-shaped bottle, then add triethylamine (10.51 mL, 75.84 mmol), slowly add chloroacetyl chloride (3.04 mL, 37.92 mmol) in DCM (10 mL) under ice bath, and naturally return to room temperature to react overnight.
  • Step 3 Add 2-chloro-N-[2-chloro-5-fluoro-4-(trifluoromethyl)phenyl]acetamide (75-c, 4.6 g, 27.59 mmol) and acetone (30 mL) to a 250 mL eggplant-shaped bottle, and then react at 50°C for 3 hours.
  • Step 4 To a 50 mL eggplant-shaped bottle, add tert-butyl 4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-5, 205.2 mg, 0.48 mmol), dioxane (10 mL), N-[2-chloro-5-fluoro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (75-d, 200 mg, 0.55 mmol) and DIPEA (0.25 mL, 1.50 mmol) and react at 80 °C for 2 hr under nitrogen protection.
  • Step 5 To a 50 mL eggplant-shaped bottle, add tert-butyl 4-(4-(2-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (75-e, 290 mg, 0.42 mmol) and solvent DCM (10 mL), add TFA (3 mL) and stir at room temperature for 2 hours.
  • Step 6 To a 25 mL eggplant-shaped bottle, add N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (75-f, 60 mg, 0.10 mmol) and 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 21.6 mg, 0.10 mmol), and add 2-methyl-2-butanol (1 mL) solution and DIPEA (0.04 mL) in sequence.
  • Step 1 Add 2-bromo-5-ethyl-[1,2,4]triazolo[1,5-a]pyrimidin-7(4H)-one (int-3a, 600 mg, 2.47 mmol), N-[2-chloro-5-fluoro-4-(trifluoromethyl)phenyl]-2-iodoacetamide (75-d, 986.9 mg, 2.71 mmol) into a 100 mL round-bottom flask, add dioxane (30 mL), DIPEA (1.23 mL, 7.40 mmol) and stir at 80 °C under nitrogen protection for 6 h.
  • Step 2 Add 2-(2-bromo-5-ethyl-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-4-yl)-N-[2-chloro-5-fluoro-4-(trifluoromethyl)phenyl]acetamide (76-a, 1.0 g, 2.01 mmol), 3,6-dihydro-2H-pyran-4-boronic acid pinacol ester (0.6 g, 3.02 mmol), Pd(dppf)Cl 2 (0.1 g, 0.20 mmol), potassium phosphate (1.4 g, 6.04 mmol) to a mixed solvent of water (5 mL) and dioxane (20 mL), raise the temperature to 85°C, and stir for 2 hours.
  • Step 3 Dissolve N-[2-chloro-5-fluoro-4-(trifluoromethyl)phenyl]-2-[2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo[1,2,4]triazolo[1,5-a]pyrimidin-4-yl]acetamide (76-b, 0.45 g, 0.9 mmol) in DMF (5 mL) solution, then slowly add NBS (0.18 g, 1.0 mmol), heat to 60 °C and stir for 3 h. Water was added to the reaction solution, extracted with ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure.
  • Step 4 2-(6-bromo-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)acetamide (76-c, 0.45 g, 0.78 mmol), (R)-1-N-Boc-2-methylpiperazine (1.56 g, 7.78 mmol) and silver tetrafluoroborate (0.18 g, 0.93 mmol) were added to DMSO (4 mL) solution in turn, vacuumed, replaced with nitrogen 3 times, heated to 120°C and stirred overnight.
  • Step 5 Add (R)-4-(4-(2-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (76-d, 0.48 g, 0.69 mmol) to a hydrochloric acid-dioxane solution (20 mL) and stir at room temperature overnight.
  • a hydrochloric acid-dioxane solution 20 mL
  • Step 6 Add 1-cyclopropyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (18-a, 82 mg, 0.42 mmol), DIPEA (32 mg, 0.25 mmol) and (R)-N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (76-e, 50 mg, 0.08 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180°C and stir for 30 min.
  • Step 1 Add 4-hydroxyisoxazole-3-carboxylic acid (int-7, 33.2 mg, 0.26 mmol), (1-chloro-2-methylprop-1-enyl)dimethylamine (35.3 mg, 0.27 mmol), and DCM (2 mL) to a 25 mL eggplant-shaped bottle, stir for 3 hours under nitrogen protection, then add DIPEA (0.14 mL, 0.86 mmol), N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (75-f, 50 mg, 0.09 mmol), and react at room temperature for 2 hours.
  • DIPEA 0.14 mL, 0.86 mmol
  • Step 1 Add 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 88 mg, 0.42 mmol), DIPEA (32 mg, 0.25 mmol) and (R)-N-(2-chloro-5-fluoro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (76-e, 50 mg, 0.08 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 Add diethyl ethoxymethylenemalonate (3-a, 0.7 g, 3.28 mmol), tetrahydro-2H-pyran-4-ylhydrazine hydrochloride (0.5 g, 3.28 mmol) and potassium carbonate (1.36 g, 9.83 mmol) to an aqueous solution (10 mL) and stir at 100°C for 6 hours. Adjust the pH to 2 with 5N HCl, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate under reduced pressure.
  • Step 2 Add 5-hydroxy-1-(tetrahydro-2H-pyran-4-yl)-1H-pyrazole-4-carboxylic acid ethyl ester (79-a, 106 mg, 0.44 mmol), DIPEA (34 mg, 0.27 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 50 mg, 0.09 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 Add 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 97 mg, 0.43 mmol), DIPEA (34 mg, 0.26 mmol) and (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (72-e, 50 mg, 0.09 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180°C and stir for 30 min.
  • Step 1 tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2a, 0.4 g, 0.6mmol), cis-2,6-dimethylmorpholine (0.35g, 3.02mmol) and potassium acetate (0.35g, 3.62mmol) were added to DMSO solution (3mL) respectively, and the mixture was stirred overnight at 120°C.
  • Step 2 Add tert-butyl 4-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(cis-2,6-dimethylmorpholinyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (81-a, 0.17 g, 0.24 mmol) to a hydrochloric acid-dioxane solution (4 mL) and stir at room temperature overnight.
  • Step 3 Add 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 70 mg, 0.34 mmol), DIPEA (26 mg, 0.2 mmol) and N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(cis-2,6-dimethylmorpholinyl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (81-b, 40 mg, 0.07 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 Add 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)- tert-butyl 2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2a, 400 mg, 0.60 mmol), 1-methyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,2,3,6-tetrahydropyridine (202 mg, 0.91 mmol) and solvent 1,4-dioxane (8 mL) and H 2 O (2 mL).
  • potassium phosphate (417 mg, 1.81 mmol) and Pd(dppf)Cl 2 (44 mg, 0.06 mmol) were added thereto and stirred at 90°C for 3 hours. 30 mL of water was added, and the mixture was extracted with ethyl acetate (30 mL ⁇ 3). The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate, and dried under reduced pressure to obtain a crude product.
  • Step 2 4-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(1-methyl-1,2,3,6-tetrahydropyridin-4-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylic acid tert-butyl ester (83-a, 300 mg, 0.44 mmol) and solvent DCM (2 mL) were added to a 100 mL eggplant-shaped bottle. TFA (0.5 mL) was then added and stirred at room temperature for 3 hours.
  • Step 3 Add 4-hydroxyisoxazole-3-carboxylic acid (int-7, 34 mg, 0.26 mmol) and solvent DCM (2 mL) to a 50 mL eggplant-shaped bottle. Then add (1-chloro-2-methylprop-1-enyl) dimethylamine (36 mg, 0.27 mmol) and stir at room temperature for 3 hours.
  • Step 1 In a 50 mL eggplant-shaped bottle, tert-butyl 4-(2-bromo-4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (int-2a, 1 g, 1.51 mmol), 2-(5,6-dihydro-4H-pyran-2-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (0.5 g, 2.26 mmol), Pd(dppf) Cl2 (0.1 g, 0.15 mmol), potassium phosphate (1.0 g, 4.53 mmol), water (5 mL) and dioxane (20 mL) were added, the temperature was raised to 85°C and stirred for 2 hours.
  • Step 2 Add tert-butyl 4-(4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)piperazine-1-carboxylate (84-a, 600 mg, 0.90 mmol) and methanolic hydrochloric acid solution (10 mL) into a 50 mL eggplant-shaped bottle and react at 50°C for 2 hours.
  • Step 4 Add 4-hydroxyisoxazole-3-carboxylic acid (36.4 mg, 0.27 mmol) and DCM (2 mL) to a 25 mL eggplant-shaped bottle, stir under nitrogen for 3 hours, then add DIPEA (0.14 mL, 0.86 mmol), N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (84-b, 50 mg, 0.09 mmol), and react at room temperature for 2 hours.
  • Step 1 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 94 mg, 0.42 mmol) and (R)-N-(5-chloro-2-methyl-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (54-e, 50 mg, 0.08 mmol) were placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added.
  • Step 1 Place N-(2-chloro-4-(trifluoromethyl)phenyl)-2-iodoacetamide (296.5 mg, 0.82 mmol) in a 100 mL eggplant-shaped bottle, add 10 mL of 1,4-dioxane solution, then add (R)-tert-butyl 4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate (int-6, 300 mg, 0.68 mmol) and DIPEA (0.34 mL, 2.04 mmol) in sequence, and stir at 75°C for 5 hours.
  • DIPEA 0.34 mL, 2.04 mmol
  • Step 2 Place (R)-4-(2-bromo-4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (86-a, 360 mg, 0.53 mmol) in a 25 mL eggplant-shaped bottle, and add 3 mL of DMSO solution. Then add morpholine (0.09 mL, 1.37 mmol) and potassium acetate (134 mg, 1.37 mmol) in sequence.
  • Step 3 Dissolve (R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-morpholinyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (86-b, 180 mg, 0.26 mmol) in 4 mL of dichloromethane solution, add 1 mL of trifluoroacetic acid and stir at room temperature for 2 hours.
  • Step 4 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 90 mg, 0.42 mmol) and (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(3-methylpiperazin-1-yl)-2-morpholinyl-7-oxo- [1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (86-c, 50 mg, 0.08 mmol) was placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added.
  • Step 1 3-(Benzyloxy)-4-oxo-4H-pyran-2-carboxylic acid (87-a, 200 mg, 0.82 mmol) was placed in a 25 mL eggplant-shaped bottle, methylamine methanol solution (3 mL) was added thereto, and heated to reflux at 80°C overnight. The reaction solution was directly concentrated to obtain crude 3-(benzyloxy)-1-methyl-4-oxo-1,4-dihydropyridine-2-carboxylic acid (87-b, 200 mg, 0.78 mmol, 95% yield). MS Calcd: 259.08; MS Found: 260.13 ([M+H] + ).
  • Step 2 Place N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (int-2, 200 mg, 0.35 mmol) and 3-(benzyloxy)-1-methyl-4-oxo-1,4-dihydropyridine-2-carboxylic acid (87-b, 137 mg, 0.69 mmol) in a 25 mL eggplant shaped bottle, add DMF solution (3 mL), and then add HATU (268 mg, 0.88 mmol) and DIPEA (0.18 mL, 1.33 mmol) in sequence, and react at room temperature overnight.
  • Step 3 2-(6-(4-(3-(benzyloxy)-1-methyl-4-oxo-1,4-dihydropyridin-2-yl)piperazin-1-yl)- 2-(3,6-Dihydro-2H-pyran-4-yl)-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-chloro-4-(trifluoromethyl)phenyl)acetamide (87-c, 70 mg, 0.09 mmol) was placed in a 25 mL eggplant-shaped bottle, DCM solution (3 mL) was added thereto, and then p-toluenesulfonic acid hydrate (82 mg, 0.43 mmol) was added, and the reaction was carried out at 50°C overnight.
  • Step 1 Add 4-hydroxyisoxazole-3-carboxylic acid (int-7, 32 mg, 0.25 mmol) and 1-chloro-N,N,2-trimethylpropyleneamine (36 mg, 0.27 mmol) to DCM solution (2 mL), stir at room temperature for 3 hours, then add N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(cis-2,6-dimethylmorpholinyl)-5-ethyl-7-oxo-6-(piperazine-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (81-b, 50 mg, 0.08 mmol) and DIPEA (65 mg, 0.5 mmol), continue to heat to 40 ° C and stir for 5 hours.
  • 4-hydroxyisoxazole-3-carboxylic acid int-7, 32 mg, 0.25 mmol
  • Step 1 Weigh 2-iodo-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (234 mg, 0.68 mmol), (R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (int-6, 300 mg, 0.68 mmol) into a 25 mL single-necked bottle, add 1,4-dioxane (5 mL) and DIEA (0.34 mL, 2.04 mmol), and react at 80 °C for 2 hours after addition.
  • 1,4-dioxane 5 mL
  • DIEA 0.34 mL, 2.04 mmol
  • Step 2 Weigh (R)-4-(2-bromo-5-ethyl-4-(2-(2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (90-a, 389 mg, 0.56 mmol), 2-(3,6-dihydro-2H-pyran-4-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (177.4 mg, 0.84 mmol), potassium phosphate (388.9 mg, 1.69 mmol), Pd(dppf)Cl 2 (82.4 mg, 0.11 mmol) was added to a 50 mL single-mouth bottle, and dioxane (5 mL) and water (2 mL) were added.
  • Step 3 Weigh (R)-4-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-4-(2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (90-b, 312 mg, 0.43 mmol) into a 50 mL single-mouth bottle, add DCM (5 mL) and trifluoroacetic acid (2 mL), and stir at room temperature for 1 hour after the addition.
  • DCM 5 mL
  • trifluoroacetic acid 2 mL
  • Step 4 Weigh 4-hydroxyisoxazole-3-carboxylic acid (int-7, 16.4 mg, 0.13 mmol) and HATU (102.7 mg, 0.27 mmol) into a 25 mL single-necked bottle, add DCM (1 mL), then add DIPEA (0.03 mL, 0.17 mmol), stir at room temperature for 5 minutes, then add (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazine-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (90-c, 50 mg, 0.08 mmol), react at room temperature for 2 hours.
  • Step 1 (R)-4-(2-bromo-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (int-6, 0.3 g, 0.68 mmol), N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)-2-iodoacetamide (294 mg, 0.82 mmol) and DIPEA (264 mg, 2.04 mmol) were added to a dioxane solution (10 mL) and stirred at 80°C for 4 hours. The reaction solution was concentrated under reduced pressure to obtain the residue.
  • Step 2 (R)-4-(2-bromo-5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (91-a, 0.2 g, 0.3 mmol), 2-(4-methoxycyclohex-1-en-1-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (307 mg, 1.29 mmol), Pd(dppf)Cl 2 ⁇ CH 2 Cl 2 (70 mg, 0.09 mmol) and potassium phosphate (274 mg, 1.29 mmol) were added to a 10 mL dioxane/water (4:1) mixed solution in sequence, and the mixture was replaced with
  • Step 3 Add (2R)-4-(5-ethyl-4-(2-((5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (91-b, 190 mg, 0.27 mmol) to a hydrochloric acid-dioxane solution (4 mL) and stir at room temperature overnight.
  • Step 4 Add 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 87 mg, 0.41 mmol), DIPEA (32 mg, 0.25 mmol) and 2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-6-((R)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(5-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (91-c, 50 mg, 0.08 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 In a 25 mL eggplant-shaped bottle, N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,4-dihydro-2H-pyran-6-yl)-5-ethyl-7-oxo-6-(piperazin-1-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (84-b, 60 mg, 0.11 mmol), 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 22.3 mg, 0.11 mmol), 2-methyl-2-butanol (2 mL) and DIPEA (0.04 mL) were added.
  • Step 1 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 115 mg, 0.51 mmol) and (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(3-methylpiperazin-1-yl)-2-morpholinyl-7-oxo- [1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (86-c, 60 mg, 0.1 mmol) was placed in a 25 mL eggplant-shaped bottle, and 2 mL of 2-methyl-2-butanol was added, followed by stirring at 180° C.
  • Step 1 Weigh 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (17-b, 85 mg, 0.40 mmol) and (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (90-c, 50 mg, 0.08 mmol) into a 25 mL single-necked bottle, add 2-methyl-2-butanol (2 mL), then add DIPEA (0.04 mL, 0.24 mmol), and after addition, heat to 180 ° C and react for 2 hours.
  • 2-methyl-2-butanol 2 mL
  • DIPEA 0.24 mmol
  • Step 1 Weigh 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 90 mg, 0.40 mmol) and (R)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(2-methyl-4-(trifluoromethyl)phenyl)acetamide (90-c, 50 mg, 0.08 mmol) into a 25 mL single-necked bottle, add 2-methyl-2-butanol (2 mL), then add DIPEA (0.04 mL, 0.24 mmol), and after addition, heat to 180 ° C and react for 2 hours.
  • 2-methyl-2-butanol 2 mL
  • DIPEA 0.24 mmol
  • Step 1 Add diethyl ethoxymethylenemalonate (3-a, 1.49 g, 6.92 mmol), phenylhydrazine hydrochloride (1.0 g, 6.92 mmol) and potassium carbonate (2.86 g, 20.75 mmol) to an aqueous solution (10 mL) and stir at 100°C for 6 hours. Adjust the pH to 2 with 5N HCl, extract with dichloromethane, wash with saturated brine, dry with anhydrous sodium sulfate, and concentrate under reduced pressure.
  • Step 2 Add 5-hydroxy-1-phenyl-1H-pyrazole-4-carboxylic acid ethyl ester (96-a, 100 mg, 0.43 mmol), DIPEA (34 mg, 0.27 mmol) and (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(3,6-dihydro-2H-pyran-4-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (14-b, 50 mg, 0.09 mmol) to 2-methyl-2-butanol solution (2 mL), heat the oil bath to 180 °C and stir for 30 min.
  • Step 1 2-(2-bromo-5-ethyl-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (56-a, 1 g, 2.1 mmol), 2-(4-methoxycyclohex-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (int-8, 760 mg, 3.2 mmol), potassium phosphate (1.4 g, 6.3 mmol) were added.
  • 1,1-bis(diphenylphosphino)diphosphine iron palladium dichloride 150 mg, 0.21 mmol was added to a mixed solvent of dioxane (15 mL) and water (4 mL), replaced with nitrogen for 3 minutes, the temperature was raised to 85 ° C, stirred for 3 hours, and concentrated under reduced pressure. The residue was poured into water and extracted with ethyl acetate (20 mL x 3). The organic layers were combined and concentrated under reduced pressure.
  • Step 2 Add 2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (98-a, 730 mg, 1.43 mmol) to DMF (10 ml), then add NBS (255 mg, 1.43 mmol), raise the temperature to 60°C and stir for 3 hours. The reaction system was then poured into water and extracted with EA (20 mL x 3). The organic layer was dried over anhydrous sodium sulfate, filtered and concentrated.
  • Step 3 2-(6-bromo-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (98-b, 510 mg, 0.87 mmol), (R)-tert-butyl 2-methylpiperazine-1-carboxylate (1.7 g, 8.7 mmol), and silver tetrafluoroborate (170 mg, 0.87 mmol) were added to dry DMSO (5 mL), and then the temperature was raised to 120° C.
  • Step 4 (2R)-4-(5-ethyl-4-(2-((3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (98-c, 410 mg, 0.58 mmol) was added to DCM (10 mL), and then trifluoroacetic acid (3 mL) was added. The mixture was stirred at room temperature for 4 hours.
  • Step 5 2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-6-((R)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (98-d, 60 mg, 0.1 mmol), DIPEA (15 mg, 0.10 mmol), 1-cyclobutyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (105 mg, 0.5 mmol) were added to 2-methyl-2-butanol (1 mL), the oil bath was heated to 180° C., stirred for 30 minutes, the reaction system was dissolved in methanol, and purified by prep-HPLC to obtain the title compound: 2-(6-((R)-4-(1-cyclobutyl-5-hydroxy-1H
  • Step 1 2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-6-((R)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)-N-(3-fluoro-2-methyl-4-(trifluoromethyl)phenyl)acetamide (98-d, 60 mg, 0.1 mmol), DIEA (15 mg, 0.10 mmol), 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 110 mg, 0.5 mmol) were added to 2-methyl-2-butanol (1 mL) and heated in an oil bath.
  • Step 1 Add (R)-4-(2-bromo-4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (86-a, 300 mg, 0.44 mmol), 4-(dimethylamino)piperidine (284 mg, 2.22 mmol) and solvent DMSO (5 mL) to a 100 mL eggplant-shaped bottle.
  • Step 2 Add (R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-2-(4-(dimethylamino)piperidin-1-yl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (101-a, 200 mg, 0.28 mmol) and solvent DCM (2 mL) to a 100 mL eggplant-shaped bottle. Then add TFA (0.5 mL) and stir at room temperature for 2 hours.
  • TFA 0.5 mL
  • Step 3 Add (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(2-(4-(dimethylamino)piperidin-1-yl)-5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (101-b, 50 mg, 0.08 mmol) and solvent 2-methyl-2-butanol (1 mL) into a 10 mL microwave tube.
  • Step 1 In a 50 mL eggplant shaped bottle, (R)-tert-butyl 4-(2-bromo-4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylate (86-a, 500 mg, 0.74 mmol), pyridin-3-ylboronic acid (136.2 mg, 1.11 mmol), Pd(dppf) Cl2 (53.6 mg, 0.07 mmol), potassium phosphate (509.7 mg, 2.22 mmol), water (5 mL) and dioxane (20 mL) were added, the temperature was raised to 85°C and stirred for 3 hours.
  • Step 2 Add (R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-2-(pyridin-3-yl)-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (103-a, 400 mg, 0.59 mmol) and methanolic hydrochloric acid solution (10 ml) into a 50 mL eggplant-shaped bottle and stir at room temperature for 2 hours.
  • Step 3 To a 25 mL eggplant-shaped bottle, add (R)-N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-6-(3-methylpiperazin-1-yl)-7-oxo-2-(pyridin-3-yl)-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (103-b, 50 mg, 0.09 mmol), 1-cyclobutyl-5-hydroxypyrazole-4-carboxylic acid ethyl ester (36.6 mg, 0.17 mmol), 2-methyl-2-butanol (2 mL), and DIPEA (0.04 mL) in a solution, evacuate and replace with nitrogen three times, and react at 180 ° C for 2 hours.
  • Step 1 Compound (R)-4-(2-bromo-4-(2-(2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (86-a, 300 mg, 0.44 mmol) and 2-(4-methoxycyclohex-1-en-1-yl)-4,4,5-5-tetramethyl-1,3,2-dioxaborolane (211 mg, 0.89 mmol) were dissolved in 1,4-dioxane (10 mL) and H 2 O (2 mL), and Pd(dppf)Cl 2 was added in sequence.
  • Step 2 Dissolve (2R)-4-(4-(2-((2-chloro-4-(trifluoromethyl)phenyl)amino)-2-oxoethyl)-5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-7-oxo-4,7-dihydro-[1,2,4]triazolo[1,5-a]pyrimidin-6-yl)-2-methylpiperazine-1-carboxylic acid tert-butyl ester (104-a, 130 mg, 0.18 mmol) in 4 mL of dichloromethane solution, add 1 mL of trifluoroacetic acid and stir at room temperature for 2 hours.
  • Step 3 Place N-(2-chloro-4-(trifluoromethyl)phenyl)-2-(5-ethyl-2-(4-methoxycyclohex-1-en-1-yl)-6-((R)-3-methylpiperazin-1-yl)-7-oxo-[1,2,4]triazolo[1,5-a]pyrimidin-4(7H)-yl)acetamide (104-b, 50 mg, 0.08 mmol) and 1-cyclopentyl-5-hydroxy-1H-pyrazole-4-carboxylic acid ethyl ester (41-a, 92 mg, 0.41 mmol) in a 25 mL eggplant-shaped bottle, and add 2 mL 2-methyl-2-butanol.

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Abstract

本发明涉及一种式(I)所示的双杂环WRN抑制剂、其制备方法及其应用。本发明还涉及包含所述化合物作为活性成分的药物组合物以及所述化合物或药物组合物用于治疗和/或预防与WRN生物学活性相关的疾病的用途。

Description

双杂环WRN抑制剂、其制备方法及其应用 技术领域
本发明属于药物化学领域，具体涉及一种双杂环WRN抑制剂、其制备方法及其应用。
背景技术
WRN是DNA解旋酶RecQ家族成员之一，参与DNA损伤修复、基因组稳定性维持、端粒维持等，WRN基因缺陷可导致Werner综合征(早老综合征)，一种以阶段性早衰和癌症易感性升高为特征的隐性遗传性疾病(Nat Rev Cancer 2003,3,169-178)。2019年，一项旨在通过基因筛选、药物敏感性和药物预测模型寻找新的肿瘤治疗靶点的项目Cancer Dependency Map(DepMap)发现WRN对于微卫星不稳定(MSI)肿瘤的生存是必需的。研究结果显示，WRN的缺失会引起MSI细胞DNA双链断裂(DSBs)、细胞周期阻滞进而引起细胞凋亡，而对于微卫星稳定(MSS)细胞则无影响(Nature 2019,568,511-516)。进一步的机制研究结果显示，WRN外切酶域功能的完好与否对MSI细胞的生存无明显影响，WRN解旋域功能的完整才是MSI细胞生存所必需的(Life Sci Alliance 4)。此外，MSI细胞中存在大规模扩张且高度不稳定的TA-二核苷酸重复序列，扩张的TA重复序列会形成非B形的十字形DNA二级结构，阻碍复制叉的延伸，WRN可解旋该结构恢复DNA的正常复制，而当WRN缺失时，十字形结构会被核酸酶MUS81切割，导致染色体基因组破坏，进而引起细胞死亡，而在MSS细胞中，TA重复序列则不会形成十字形结构，所以WRN缺失会特异性地导致MSI细胞死亡(Nature 2020,586,292-298)。因此，靶向WRN解旋域的抑制剂可能是治疗MSI癌症的有效措施。
目前暂未发现WRN靶点抑制相关的药物上市，基于WRN靶点抑制在微卫星不稳定(MSI)肿瘤治疗中潜在的医学价值，开发WRN靶点抑制相关的化合物具有十分重要的社会意义和医学价值。
发明内容
发明要解决的问题：
尽管目前已公开了一些WRN抑制剂的专利申请，然而，基于目前尚无WRN靶点药物上市及WRN靶点抑制在微卫星不稳定(MSI)肿瘤治疗中潜在医学价值，新的化合物仍需进一步开发。经过不断努力，本申请发明人设计了具有通式(I)所示的结构的化合物，并发现具有此类结构的化合物表现出优异的WRN抑制效果和作用，可用于治疗MSI肿瘤相关疾病。
用于解决问题的方案：
为了解决上述问题，本申请发明人已经尝试进行了深入研究并发现式(I)所示双环结构化合物及其衍生物能达到期望的目的，结果完成本发明。
本发明保护以下具体实施方案：
一种式(I)所示的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：
其中，
R1选自卤素、C1-6烷基、C2-6烯基、C2-6炔基、5元或6元环烯基、5-10元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所述烷基、烯基、炔基、环烯基、杂环基、杂芳基、苯基各自独立地是未取代的或被1个或多个相同或不同的Rg取代；
每个Rg各自独立地选自：羟基、卤素、C1-4烷基、C1-4烷氧基、HOC(O)-(CH2)n-、H3C-C(O)(CH2)n-、C1-4烷基-O-C(O)(CH2)n-、＝O、氮杂环丁基、吡咯烷基、R’R”N-，其中，R’、R”各自独立地选自H、C1-4烷基、C1-4卤代烷基，n＝0，1或2；所述氮杂环丁基或吡咯烷基通过N原子连接到R1主基团上；所述C1-4烷基、C1-4烷氧基、氮杂环丁基、吡咯烷基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
R2选自 其中，L选自7-10元螺杂环烷基、所述螺杂环烷基含有2-3个N原子；
R3选自环丙基、甲氧基、-SCH3、C1-4烷基，所述烷基是未取代的或被1个或多个选自羟基或卤素的取代基取代；
R4选自H、卤素、羟基、氰基、C1-4烷基、C3-5环烷基、C1-4烷氧基，所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
R5选自H、甲氧基、羟基、氰基、甲基、卤素；
R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个或多个卤素取代；
R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基、C1-4烷氧基；所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取 代；
X选自N或CR8，R8为H或卤素；
R2a选自6-14元芳基、5-14元杂芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述芳基、杂芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rc取代；
R2b选自5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环烷基、5-14元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个选自N、O或S的杂原子；所述5-14元杂环烷基、5-14元杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环烷基、5-14元杂环烯基各自独立地被一个或多个Rd取代；
R2e选自5-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述杂芳基、芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rf取代；
每个Rc各自独立地选自卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基、氰基、氨基；
每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、C1-6卤代烷氧基、C3-6卤代环烷基、羟基、卤素、4-7元杂环烷基、5元或6元杂芳基、苯基；所述4-7元杂环烷基含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代；
每个Rf各自独立地选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基。
一种式(I)所示化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：
其中，
R1选自卤素、C1-6烷基、C2-6烯基、C2-6炔基、5元或6元环烯基、5元或6元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基含有1-2个选自N、O或S的杂原子，所述烷基、烯基、炔基、环烯基、杂环基、杂芳基、苯基是未取代的或被1个或多个相同或不同的Rg取代；
每个Rg各自独立地选自：羟基、卤素、C1-4烷基、C1-4烷氧基、HOC(O)-(CH2)n-、H3C-C(O)(CH2)n-、C1-4烷基-O-C(O)(CH2)n-、＝O、氮杂环丁基、吡咯烷基、R’R”N-，其中，R’、R”各自独立地选自H、C1-4烷基、C1-4卤代烷基，n＝0，1或2；所述氮杂环丁基或吡咯烷基通过N原子连接到R1主基团上；所述C1-4烷基、C1-4烷氧基、氮杂环丁基、吡咯烷基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
R2选自其中，L选自7-10元螺杂环基、
R3选自环丙基、甲氧基、-SCH3、C1-4烷基，所述烷基是未取代的或被1个或多个选自羟基或卤素的取代基取代；
R4选自H、卤素、羟基、氰基、C1-4烷基、C3-5环烷基、C1-4烷氧基，所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
R5选自H、甲氧基、羟基、氰基、甲基、卤素；
R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个或多个卤素取代；
R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基、C1-4烷氧基；所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
X选自N或CR8，R8为H或卤素；
R2a选自6-14元芳基、5-14元杂芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述芳基、杂芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rc取代；
R2b选自5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基各自独立地被一个或多个Rd取代；
R2e选自5-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述杂芳基、芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rf取代；
每个Rc各自独立地选自卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基、氰基、氨基；
每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、C1-6卤代烷氧基、C3-6卤代环烷基、4-7元杂环烷基、羟基或卤素；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；
每个Rf各自独立地选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基。
一种式(I)所示化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：
其中，
R1选自卤素、C1-6烷基、C2-6烯基、C2-6炔基、5元或6元环烯基、5元或6元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基含有1-2个选自N、O或S的杂原子，所述烷基、烯基、炔基、环烯基、杂环基、杂芳基、苯基是未取代的或被1个或多个相同或不同的Rg取代；
每个Rg各自独立地选自：羟基、卤素、C1-4烷基、C1-4烷氧基、HOC(O)-(CH2)n-、H3C-C(O)(CH2)n-、C1-4烷基-O-C(O)(CH2)n-、＝O、氮杂环丁基、吡咯烷基、R’R”N-，其中，R’、R”各自独立地选自H、C1-4烷基、C1-4卤代烷基，n＝0，1或2；所述氮杂环丁基或吡咯烷基通过N原子连接到R1主基团上；所述C1-4烷基、C1-4烷氧基、氮杂环丁基、吡咯烷基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
R2选自其中，L选自7-10元螺杂环基、
R3选自环丙基、甲氧基、-SCH3、C1-4烷基，所述烷基是未取代的或被1个或多个选自羟基或卤素的取代基取代；
R4选自H、卤素、羟基、氰基、C1-4烷基、C3-5环烷基、C1-4烷氧基，所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
R5选自H、甲氧基、羟基、氰基、甲基、卤素；
R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个或多个卤素取代；
R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基、C1-4烷氧基；所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
X选自N或CR8，R8为H或卤素；
R2a选自6-14元芳基、5-14元杂芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述芳基、杂芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rc取代；
R2b选自5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基各自独立地被一个或多个Rd取代；
R2e选自5-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述杂芳基、芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rf取代；
每个Rc各自独立地选自卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基、氰基、氨基；
每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、C1-6卤代烷氧基、C3-6卤代环烷基、羟基或卤素；
每个Rf各自独立地选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基。
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：R2选自
在一个具体实施方式中，R2选自
在一个具体实施方式中，R2选自
在一个具体实施方式中，R2选自
在一个具体实施方式中，R2选自
在一个具体实施方式中，R2选自
在一个具体实施方式中，R2选自
在一个具体实施方式中，R2选自
在一个具体实施方式中，R2b选自5元杂芳基、8-14元杂芳基；所述5元杂芳基、8-14元杂芳基各自独立地含有1-4个选自N、O和S的杂原子；所述5元杂芳基、8-14元杂芳基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自5元杂芳基、8元杂芳基、9元杂芳基、10元杂芳基；所述5元杂芳基、8元杂芳基、9元杂芳基、10元杂芳基各自独立地含有1-4个选自N、O和S的杂原子；所述5元杂芳基、8元杂芳基、9元杂芳基、10元杂芳基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自5元杂芳基、5元杂芳基并5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基、6元杂芳基并6元杂芳基；所述5元杂芳基、6元杂芳基各自独立地含有1-2个选自N、O和S的杂原子；所述5元杂芳基、5元杂芳基并5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基、6元杂芳基并6元杂芳基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基；所述5元杂芳基、6元杂芳基各自独立地含有1-2个选自N、O和S的杂原子；所述5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自吡咯基、呋喃基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基、苯并吡咯基、苯并呋喃基、苯并噻吩基、苯并吡唑基、苯并咪唑基、苯并噻唑基、苯并异噻唑基、苯并噁唑基、苯并异噁唑基、吡啶并吡咯基、吡啶并呋喃基、吡啶并噻吩基、吡啶并吡唑基、吡啶并咪唑基、吡啶并噻唑基、吡啶并异噻唑基、吡啶并噁唑基、吡啶并异噁唑基、苯并吡啶基、苯并嘧啶基；所述吡咯基、呋喃基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基、苯并吡咯基、苯并呋喃基、苯并噻吩基、苯并吡唑基、苯并咪唑基、苯并噻唑基、苯并异噻唑基、苯并噁唑基、苯并异噁 唑基、吡啶并吡咯基、吡啶并呋喃基、吡啶并噻吩基、吡啶并吡唑基、吡啶并咪唑基、吡啶并噻唑基、吡啶并异噻唑基、吡啶并噁唑基、吡啶并异噁唑基、苯并吡啶基、苯并嘧啶基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自吡唑基、苯并吡啶基、异噁唑基、异噻唑基、吡啶并吡唑基、吡啶并咪唑基、吡啶并呋喃基、吡啶并吡咯基、吡啶并噻吩基；所述吡唑基、苯并吡啶基、异噁唑基、异噻唑基、吡啶并吡唑基、吡啶并咪唑基、吡啶并呋喃基、吡啶并吡咯基、吡啶并噻吩基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自 所述 各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自
在一个具体实施方式中，每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、羟基、卤素，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1或2个选自N、O和S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O和S的杂原子。
在一个具体实施方式中，每个Rd各自独立地选自C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、羟基、卤素，其中有且只有一个Rd 为羟基；所述4-7元杂环烷基含有1或2个选自N、O和S的杂原子；所述5元或6元杂芳基各自独立地含有1个、2个或3个选自N、O和S的杂原子。
在一个具体实施方式中，每个Rd各自独立地选自C1-4烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C1-4烷氧基、4-7元杂环烷基、6元杂芳基、苯基、羟基、卤素，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1个或2个O杂原子；所述6元杂芳基含有1个或2个N杂原子。
在一个具体实施方式中，每个Rd各自独立地选自C1-2烷基、C3-4烷基、C3-4环烷基、C5-6环烷基、C1-2氟代烷基、C3-4氟代烷基、C1-2烷氧基、C3-4烷氧基、4-7元杂环烷基、吡啶基、嘧啶基、哒嗪基、吡嗪基、苯基、羟基、卤素，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1个O杂原子。
在一个具体实施方式中，每个Rd各自独立地选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基、环丙基、环丁基、环戊基、环己基、氟代甲基、氟代乙基、氟代丙基、氟代异丙基、甲氧基、乙氧基、4-7元杂环烷基、吡啶基、羟基、F、Cl、Br、苯基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1个O杂原子。
在一个具体实施方式中，每个Rd各自独立地选自甲基、异丙基、叔丁基、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、 Cl、苯基、羟基，其中有且只有一个Rd为羟基。
在一个具体实施方式中，上述所述羟基取代在R2b与主基团连接位点的邻位或间位的碳原子上。
在一个具体实施方式中，上述所述羟基取代在R2b与主基团连接位点的邻位的碳原子上。
在一个具体实施方式中，R2b选自
在一个具体实施方式中，每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、卤素； 所述4-7元杂环烷基含有1或2个选自N、O和S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O和S的杂原子。
在一个具体实施方式中，每个Rd各自独立地选自C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、卤素；所述4-7元杂环烷基含有1或2个选自N、O和S的杂原子；所述5元或6元杂芳基各自独立地含有1个、2个或3个选自N、O和S的杂原子。
在一个具体实施方式中，每个Rd各自独立地选自C1-4烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C1-4烷氧基、4-7元杂环烷基、6元杂芳基、苯基、卤素；所述4-7元杂环烷基含有1个或2个O杂原子；所述6元杂芳基含有1个或2个N杂原子。
在一个具体实施方式中，每个Rd各自独立地选自C1-2烷基、C3-4烷基、C3-4环烷基、C5-6环烷基、C1-2氟代烷基、C3-4氟代烷基、C1-2烷氧基、C3-4烷氧基、4-7元杂环烷基、吡啶基、嘧啶基、哒嗪基、吡嗪基、苯基、卤素；所述4-7元杂环烷基含有1个O杂原子。
在一个具体实施方式中，每个Rd各自独立地选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基、环丙基、环丁基、环戊基、环己基、氟代甲基、氟代乙基、氟代丙基、氟代异丙基、甲氧基、乙氧基、4-7元杂环烷基、吡啶基、F、Cl、Br、苯基；所述4-7元杂环烷基含有1个O杂原子。
在一个具体实施方式中，每个Rd各自独立地选自甲基、异丙基、叔丁基、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、 Cl、苯基。
在一个具体实施方式中，R2b选自
在一个具体实施方式中，R2b选自5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环烷基、5-10元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个选自N、O或S的杂原子；所述5-10元杂环烷基、5-10元杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环烷基、5-10元杂环烯基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个选自N、O或S的杂原子；所述杂环烷基、杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自5元杂芳基、8-10元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个选自N、O或S的杂原子；所述杂环烷基、杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-10元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基各自独立地被一个或多个Rd取代。
在一个具体实施方式中，R2b选自5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、5元或6元杂环烷基并5元或6元杂环烯基；所述杂环烷基、杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、5元或6元杂环烷基并5元或6元杂环烯基各自独立地被1个、2个、3个或4个Rd取代。
在一个具体实施方式中，R2b选自5元杂芳基、苯基并5元或6元杂芳基、6元杂芳基并5元杂芳基、1,5-二氢-2H-吡咯-2-酮基、吡啶酮基、6元杂环烷基并 吡啶酮基，所述杂芳基、杂环烷基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、6元杂芳基并5元杂芳基、1,5-二氢-2H-吡咯-2-酮基、吡啶酮基、6元杂环烷基并吡啶酮基各自独立地被1个、2个、3个或4个Rd取代。
在一个具体实施方式中，R2b选自异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 所述异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 各自独立地被1个、2个、3个或4个Rd取代。
在一个具体实施方式中，R2b选自异噁唑基、吡唑基、异噻唑基、所述异噁唑基、吡唑基、异噻唑基、各自独立地被1个、2个、3个或4个Rd取代。
在一个具体实施方式中，R2b选自所述各自独立地被1个、2个、3个或4个Rd取代。
在一个具体实施方式中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氨基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1个、2个或3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1- 3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自卤素、C1-4烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C1-4烷氧基、4-7元杂环烷基、苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、氰基或羟基；所述4-7元元杂环烷基各自独立地含有1个O杂原子；所述苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基是未取代的或被一个或多个选自F、Cl、氰基、甲基、乙基、丙基、环丙基、环丁基、环戊基、甲氧基、乙氧基、丙氧基、氟代甲基、氟代甲氧基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、氟取代的苯基、氯取代的苯基、氰基取代的苯基、甲基取代的苯基、甲氧基取代的苯基、三氟甲基取代的苯基、三氟甲氧基取代的苯基、氰基取代的吡啶基、氟取代的吡啶基、氯取代的吡啶基、甲基取代的吡啶基、甲氧基取代的吡啶基、甲基取代的吡唑基、三氟甲基取代的吡啶基、三氟甲氧基取代的吡啶基、氰基取代的吡啶基或羟基。
在一个具体实施方式中，每个Rd各自独立地选自苯基、噻吩基、吡啶基、羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl。
在一个具体实施方式中，每个Rd各自独立地选自C1-6烷基、C3-6环烷基、 C1-6卤代烷基、氰基、氨基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、羟基或卤素，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C1-4卤代烷基、C5-6卤代烷基、C3-4环烷基、C5-6环烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1个、2个或3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3- 5环烷基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自卤素、C1-4烷基、C1-4卤代烷基、C3-4环烷基、C5-6环烷基、C1-4烷氧基、4-7元杂环烷基、苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1个O杂原子；所述苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基是未取代的或被一个或多个选自F、Cl、氰基、甲基、乙基、丙基、环丙基、环丁基、环戊基、甲氧基、乙氧基、丙氧基、氟代甲基、氟代甲氧基的取代基取代。
在一个具体实施方式中，每个Rd各自独立地选自苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、氟取代的苯基、氯取代的苯基、氰基取代的苯基、甲基取代的苯基、甲氧基取代的苯基、三氟甲基取代的苯基、三氟甲氧基取代的苯基、氰基取代的吡啶基、氟取代的吡啶基、氯取代的吡啶基、甲基取代的吡啶基、甲氧基取代的吡啶基、甲基取代的吡唑基、三氟甲基取代的吡啶基、三氟甲氧基取代的吡啶基、氰基取代的吡啶基或羟基，其中有且只有一个Rd为羟基。
在一个具体实施方式中，每个Rd各自独立地选自苯基、噻吩基、吡啶基、羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl，其中有且只有一个Rd为羟基。
在一个具体实施方式中，每个Rd各自独立地选自苯基、噻吩基、吡啶基、羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl，其中有且只有一个Rd为羟基，且羟基取代在R2b与主基团连接位点的邻位。
在一个具体实施方式中，R2b选自
在一个具体实施方式中，R2b选自
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：R2选自
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：R2选自
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：R2选自
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：R2选自
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的 盐：R2b选自5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基各自独立地被一个或多个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基；所述杂芳基、杂环基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、苯基并5元或6元杂芳基、6元杂芳基并5元杂芳基、5元杂环基、6元杂环基、6元杂环基并吡啶酮基，所述杂芳基、杂环基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并6元杂芳基、6元杂芳基并5元杂芳基、5元杂环基、6元杂环基、6元杂环基并吡啶酮基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 所述异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：每 个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、C1-6烷氧基、4-7元杂环烷基、氨基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、氰基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C3-6环烷基、C1-4卤代烷基、C1-4烷氧基、4-7元杂环烷基、氰基或羟基；所述4-7元杂环烷基各自独立地含有1个O杂原子。
在一些具体实施方案中，每个Rd各自独立地选自甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、或羟基。
在一些具体实施方案中，每个Rd各自独立地选自羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl。
在一些具体实施方案中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1- 6烷氧基、4-7元杂环烷基、羟基或卤素，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、氰基或羟基，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C1-4卤代烷基、C5-6卤代烷基、C3-4环烷基、C5-6环烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂 环烷基各自独立地含有1或2个选自N、O或S的杂原子。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C1-4卤代烷基、C3-6环烷基、C1-4烷氧基、4-7元杂环烷基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1个O杂原子。
在一些具体实施方案中，每个Rd各自独立地选自甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、或羟基，其中有且只有一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl，其中有且只有一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl，其中有且只有一个Rd为羟基，且羟基取代在R2b与主基团连接位点的邻位。
在一些具体实施方案中，R2b选自
在一些具体实施方案中，
在一些具体实施方案中，R2选自
在一些具体实施方案中，R2选自
在一些具体实施方案中，R2选自
在一些具体实施方案中，R2b选自5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基各自独立地被一个或多个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基；所述杂芳基、杂环基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自5元杂芳基、苯基并6元杂芳基、6元杂芳基并5元杂芳基、6元杂环基、6元杂环基并吡啶酮基，所述杂芳基、杂环基 各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并6元杂芳基、6元杂芳基并5元杂芳基、6元杂环基、6元杂环基并吡啶酮基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自吡唑基、噻唑基、噻吩基、异噻唑基、苯基并吡啶基、吡啶并吡咯基、吗啉基并吡啶酮基、吡啶酮基；所述吡唑基、噻唑基、噻吩基、异噻唑基、苯基并吡啶基、吡啶并吡咯基、吗啉基并吡啶酮基、吡啶酮基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基或羟基。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、氰基或羟基。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C3-6环烷基、C1-4卤代烷基、氰基或羟基。
在一些具体实施方案中，每个Rd各自独立地选自甲基、乙基、丙基、F、Cl、Br、环丙基、环丁基、环戊基、三氟甲基或羟基。
在一些具体实施方案中，每个Rd各自独立地选自甲基、Cl、Br、环丙基或羟基。
在一些具体实施方案中，每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、羟基或卤素，其中至少一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基或羟基，其中至少一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基或羟基，其中有且只有一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C1-4卤代烷基、C5-6卤代烷基、C3-4环烷基、C5-6环烷基、氰基或羟基，其中有且只有一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自卤素、C1-4烷基、C1-4卤代烷基、C3-6环烷基、氰基或羟基，其中有且只有一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自甲基、乙基、丙基、F、Cl、Br、环丙基、环丁基、环戊基、三氟甲基或羟基，其中有且只有一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自甲基、Cl、Br、环丙基或羟基，其中有且只有一个Rd为羟基。
在一些具体实施方案中，每个Rd各自独立地选自甲基、Cl、Br、环丙基或羟基，其中有且只有一个Rd为羟基，且羟基取代在R2b与主基团连接位点的邻位。
在一些具体实施方案中，R2b选自
各自独立地可进一步被1个、2个或3个Rd取代。
在一些具体实施方案中，R2b选自
在一些具体实施方案中，R2b选自5元杂芳基，所述5元杂芳基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自吡唑基、噻唑基、噻吩基、异噻唑基，所述吡唑基、噻唑基、噻吩基、异噻唑基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自 可进一步被1个、2个或3个Rd取代。
在一些具体实施方案中，R2b选自可进一步被1个、2个或3个Rd取代。
在一些具体实施方案中，R2b选自：
在一些具体实施方案中，R2b选自：
在一些具体实施方案中，R2b选自6元杂环基、6元杂环基并6元杂环基，所述杂环基含有1-2个选自N、O或S的杂原子；所述6元杂环基、6元杂环基并6元杂环基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自吗啉基并吡啶酮基、吡啶酮基；所述吗啉基并吡啶酮基、吡啶酮基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自吡啶酮基；所述吡啶酮基被1个、2个、3 个或4个Rd取代。
在一些具体实施方案中，R2b选自2-吡啶酮基；所述2-吡啶酮基被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自可进一步被1个、2个或3个Rd取代。
在一些具体实施方案中，R2b选自可进一步被1个、2个或3个Rd取代。
在一些具体实施方案中，R2b选自
在一些具体实施方案中，R2b选自
在一些具体实施方案中，R2b选自苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基；所述杂芳基含有1-2个选自N、O或S的杂原子；所述苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自苯基并6元杂芳基、6元杂芳基并5元杂芳基，所述杂芳基含有1-2个选自N、O或S的杂原子；所述苯基并6元杂芳基、6元杂芳基并5元杂芳基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自苯基并吡啶基、吡啶并吡咯基；所述苯基并吡啶基、吡啶并吡咯基各自独立地被1个、2个、3个或4个Rd取代。
在一些具体实施方案中，R2b选自 可进一步被1个、2个或3个Rd取代。
在一些具体实施方案中，R2b选自
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：R1选自卤素、C1-6烷基、5元或6元环烯基、5-10元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所 述烷基、环烯基、杂环基、杂芳基、苯基各自独立地是未取代的或被1个或多个Rg取代。
在一个具体实施方式中，R1选自5元或6元环烯基、5-10元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所述环烯基、杂环基、杂芳基、苯基各自独立地是未取代的或被1个或多个Rg取代。
在一个具体实施方式中，R1选自5元或6元环烯基、5-10元杂环烷基、5元或6元杂环烯基、5元或6元杂芳基、苯基，所述杂环烷基、杂环烯基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所述环烯基、杂环烷基、杂环烯基、杂芳基、苯基各自独立地是未取代的或被1个或多个Rg取代。
在一个具体实施方式中，R1选自6元环烯基、6-9元杂环烷基、6元杂环烯基、吡啶基、嘧啶基、哒嗪基，所述杂环烷基、杂环烯基各自独立地含有1-2个选自N、O或S的杂原子，所述环烯基、杂环烷基、杂环烯基各自独立地是未取代的或被1个或多个Rg取代。
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：每个Rg各自独立地选自羟基、卤素、C1-4烷基、C1-4烷氧基、＝O、R’R”N-；所述烷基、烷氧基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；其中，R’、R”各自独立地选自H、C1-4烷基、C1-4卤代烷基。
在一个具体实施方式中，每个Rg各自独立地选自羟基、卤素、C1-2烷基、C3- 4烷基、C1-2烷氧基、C3-4烷氧基、＝O、R’R”N-；所述烷基、烷氧基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；其中，R’、R”各自独立地选自H、C1-2烷基、C3-4烷基、C1-2卤代烷基、C3-4卤代烷基。
在一个具体实施方式中，每个Rg各自独立地选自羟基、F、Cl、C1-2烷基、C1-2烷氧基、＝O、R’R”N-；所述烷基、烷氧基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；其中，R’、R”各自独立地选自H、C1-2烷基、C1-2卤代烷基。
在一个具体实施方式中，每个Rg各自独立地选自羟基、F、甲基、甲氧基、＝O、R’R”N-；所述甲基是未取代的或被1或多个选自F、Cl、羟基、甲氧基的取代基取代；其中，R’、R”各自独立地选自H、甲基。
在一个具体实施方式中，上述化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：R1选自
在一个具体实施方式中，R1选自
在一个具体实施方式中，R1选自
在一些具体实施方案中，R1选自卤素、C1-6烷基、5元或6元杂环基，所述杂环基含有1-2个选自N、O或S的杂原子，所述烷基、杂环基是未取代的或被1个或多个Rg取代。
在一些具体实施方案中，R1选自5元或6元杂环基，所述杂环基含有1-2个选自N、O或S的杂原子，所述杂环基是未取代的或被1个或多个Rg取代。
在一些具体实施方案中，R1选自5元或6元杂环烷基、5元或6元杂环烯基，所述杂环烷基、杂环烯基含有1-2个选自N、O或S的杂原子，所述杂环烷基、杂环烯基是未取代的或被1个或多个Rg取代。
在一些具体实施方案中，R1选自5元或6元杂环烯基，所述杂环烯基含有1-2个选自N或O的杂原子，所述杂环烯基是未取代的或被1个或多个Rg取代。
在一些具体实施方案中，R1选自 是未取代的或被1个或多个Rg取代。
在一些具体实施方案中，每个Rg各自独立地选自羟基、卤素、C1-4烷基、C1- 4烷氧基；所述C1-4烷基、C1-4烷氧基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代。
在一些具体实施方案中，每个Rg各自独立地选自羟基、卤素、C1-2烷基、C3- 4烷基、C1-2烷氧基、C3-4烷氧基；所述烷基、烷氧基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代。
在一些具体实施方案中，每个Rg各自独立地选自羟基、F、Cl、C1-2烷基、C1-2烷氧基；所述烷基、烷氧基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代。
在一些具体实施方案中，每个Rg各自独立地选自羟基、F、甲基、乙基；所述甲基、乙基是未取代的或被1或多个选自卤素、羟基、甲氧基的取代基取代。
在一些具体实施方案中，R1选自
在一些具体实施方案中，R1选自
在一些具体实施方案中，R1选自
在一些具体实施方案中，R3选自C1-4烷基，所述烷基是未取代的或被1个、2个或3个选自羟基或卤素的取代基取代。
在一些具体实施方案中，R3选自C1-4烷基，所述烷基是未取代的。
在一些具体实施方案中，R3选自C1-2烷基，所述烷基是未取代的。
在一些具体实施方案中，R3选自甲基或乙基。
在一些具体实施方案中，R3选自乙基。
在一些具体实施方案中，R4选自H、卤素、C1-4烷基，所述烷基是未取代的或被1个、2个或3个卤素取代。
在一些具体实施方案中，R4选自H、卤素、C1-4烷基，所述烷基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R4选自H、F、Cl、Br、C1-4烷基，所述C1-4烷基是未取代的。
在一些具体实施方案中，R4选自H、F、Cl、Br、C1-2烷基，所述C1-2烷基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R4选自H、F、Cl、Br、甲基、乙基，所述甲基、乙基各自独立地是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R4选自H、F、Cl、Br、甲基、-CF3。
在一些具体实施方案中，R4选自Cl、甲基、-CF3。
在一些具体实施方案中，R4选自H、F、Cl、Br、甲基、乙基，所述甲基、乙基是未取代的。
在一些具体实施方案中，R4选自H、F、Cl、Br或甲基。
在一些具体实施方案中，R4选自Cl或甲基。
在一些具体实施方案中，R5选自H、甲基、卤素。
在一些具体实施方案中，R5选自H、F、Cl或甲基。
在一些具体实施方案中，R5选自H、Cl或甲基。
在一些具体实施方案中，R5选自H。
在一些具体实施方案中，R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个、2个或3个卤素取代。
在一些具体实施方案中，R6选自H、卤素、C1-4烷基，所述烷基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R6选自H、卤素、C1-2烷基，所述烷基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R6选自H、F、Cl、甲基、乙基，所述甲基、乙基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R6选自H、F、Cl、甲基或-CF3。
在一些具体实施方案中，R6选自H、F或Cl。
在一些具体实施方案中，R6选自H、卤素、C1-4烷基，所述烷基被1个、2个或3个F取代。
在一些具体实施方案中，R6选自H、卤素、C1-2烷基，所述烷基被1个、2个或3个F取代。
在一些具体实施方案中，R6选自H、F、Cl、甲基、乙基，所述甲基、乙基被1个、2个或3个F取代。
在一些具体实施方案中，R6选自H、F、Cl或-CF3。
在一些具体实施方案中，R6选自H、F或Cl。
在一些具体实施方案中，R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基；所述烷基、环烷基是未取代的或被1个或多个卤素取代。
在一些具体实施方案中，R7选自-SF5、-C(O)H、卤素、C1-4烷基；所述烷基是未取代的或被1个、2个或3个卤素取代。
在一些具体实施方案中，R7选自-SF5、-C(O)H、卤素、C1-4烷基；所述烷基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R7选自-SF5、-C(O)H、卤素、C1-2烷基；所述烷基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R7选自-SF5、-C(O)H、F、Cl、Br、甲基、乙基；所述甲基、乙基是未取代的或被1个、2个或3个F取代。
在一些具体实施方案中，R7选自-CF3、-CHF2、-CH2CH3、Cl、Br、-SF5或-C(O)H。
在一些具体实施方案中，R7选自-CF3。
在一些具体实施方案中，R7选自-OCF3。
在一些具体实施方案中，X选自N或CR8，R8选自H、F、Cl或Br。
在一些具体实施方案中，R8选自H或F。
在一些具体实施方案中，功能基团选自
在一些具体实施方案中，功能基团选自
在一些具体实施方案中，功能基团选自
在一些具体实施方案中，功能基团选自
在一些具体实施方案中，功能基团选自
在一些具体实施方案中，式(I)具有选自式(IB)所示通式结构：
其中，R1、R2b、R3、R4各自定义如式(I)中所述定义。
在一些具体实施方案中，式(I)具有式(IB-1)所示结构：
其中，R1、R2b、R4定义如式(I)中所述定义。
在一些具体实施方案中，式(I)具有选自式(IIA)所示结构：
其中，R1、R2b、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIA)具有式(IIA-1)所示结构：
其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIA-1)具有式(IIA-1-1)所示结构：
其中，R1、Rd定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIA)具有式(IIA-2)所示结构：
其中，R1、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIA-2)具有式(IIA-2-1)所示结构：
其中，R1定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIA)具有式(IIA-3)所示结构：
其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIA-3)具有式(IIA-3-1)所示结构：
其中，R1、Rd定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIA)具有式(IIA-4)所示结构：
其中，环B为5元杂芳基；所述5元杂芳基含有1-2个选自N、O和S的杂原子；所述5元杂芳基为未取代的或被1个或多个C1-4烷基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B为5元杂芳基；所述5元杂芳基含有1个选自N、O和S的杂原子；所述5元杂芳基为未取代的或被1个或多个C1-4烷基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基；所述呋喃基、吡咯基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基；所述呋喃基、吡咯基、噻吩基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基；所述呋喃基、吡咯基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基；所述吡咯基为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、 异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基；所述吡咯基为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基；所述吡咯基为未取代的或被1个甲基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基；所述吡咯基为未取代的或被1个甲基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，式(I)具有选自式(IIB)所示结构：
其中，R1、R2b、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIB)具有式(IIB-1)所示结构：
其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIB-1)具有式(IIB-1-1)所示结构：
其中，R1、Rd定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIB)具有式(IIB-2)所示结构：
其中，R1、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIB-2)具有式(IIB-2-1)所示结构：
其中，R1定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIB)具有式(IIB-3)所示结构：
其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIB-3)具有式(IIB-3-1)所示结构：
其中，R1、Rd定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIB)具有式(IIB-4)所示结构：
其中，环B为5元杂芳基；所述5元杂芳基含有1-2个选自N、O和S的杂原子；所述5元杂芳基为未取代的或被1个或多个C1-4烷基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B为5元杂芳基；所述5元杂芳基含有1个选自N、O和S的杂原子；所述5元杂芳基为未取代的或被1个或多个C1-4烷基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基；所述呋喃基、吡咯基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基；所述呋喃基、吡咯基、噻吩基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基；所述呋喃基、吡咯基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基；所述吡咯基为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基；所述吡咯基为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基、噻吩基；所述吡咯基为未取代的或被1个甲基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，环B选自呋喃基、吡咯基；所述吡咯基为未取代的或被1个甲基取代；X、R4、R5、R6、R7定义如前所述。
在一些具体实施方案中，式(I)具有选自式(IIC)所示结构：
其中，R1、R2b、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIC)具有式(IIC-1)所示结构：
其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIC-1)具有式(IIC-1-1)所示结构：
其中，R1、Rd定义如式(I)中所述定义。
在一些具体实施方案中，所述式(IIC)具有式(IIC-2)所示结构：
其中，R1、R4、R5、R6、R7定义如 式(I)中所述定义。
在一些具体实施方案中，所述式(IIC-2)具有式(IIC-2-1)所示结构：
其中，R1定义如式(I)中所述定义。
在一些具体实施方案中，式(I)选自下述具体化合物：









术语“药学上可接受的盐”指本发明化合物的盐，由本发明发现的具有特定取代基的化合物与相对无毒的酸或碱制备。当本发明的化合物中含有相对酸性的功能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的碱与这类化合物接触的方式获得碱加成盐。当本发明的化合物中含有相对碱性的官能团时，可以通过在纯的溶液或合适的惰性溶剂中用足够量的酸与这类化合物接触的方式获得酸加成盐。
术语“前药”是指本发明发现具有特定取代基的式(I)所示化合物的衍生物，它们自身可能具有较弱的活性甚至没有活性，但是在给药后，在生理条件下(例如通过代谢、溶剂分解或另外的方式)被转化成本发明发现的具有特定取代基的化合物，在体内产生相应的生物活性。
术语“代谢物”是指本发明发现具有特定取代基的式(I)所示化合物在体内通过代谢作用所得到的产物。一个化合物的代谢产物可以通过所属领域公知的技术来进行鉴定，其活性可以通过如本发明所描述的那样采用试验的方法进行表征。这样的产物可以是通过给药化合物经过氧化，还原，水解，酰氨化，脱酰氨作用，酯化，脱脂作用，酶裂解等等方法得到。相应地，本发明包括化合物的代谢产物，包括将本发明的化合物与哺乳动物充分接触一段时间所产生的代谢产物。
术语“氘代化合物”指的是本发明化合物包括至少一个氘原子，具体是指本发明化合物中的一个或多个氢原子可以被氘原子置换或取代。在一些实施方案中，该化合物包括两个或更多个氘原子。在一些实施方案中，该化合物包括1、2、3、4、5、6、7、8、9、10、11或12个氘原子。用于将同位素包括到有 机化合物中的合成方法在本领域中为已知的。
制备方法：
本发明还提供制备所述化合物的方法。本发明通式(I)所述化合物的制备可通过以下示例性方法和实施例完成，但这些方法和实施例不应以任何方式被认为是对本发明范围的限制。也可通过本领域技术人员所知的合成技术合成本发明所述的化合物，或者综合使用本领域已知的合成方法和本发明所述方法。每步反应所得的产物用本领域已知的分离技术得到，包括但不限于萃取、过滤、蒸馏、结晶、色谱分离等。合成所需的起始原料和化学试剂可以根据文献常规合成(如Scifinder提供的)或购买。
本发明通式(I)所述化合物可按照以下方法所述路线合成：
方案一：
1)将A与C进行取代反应得到中间体D；
2)脱去保护基得到中间体E；
3)中间体E与相应的羧酸缩合得到产物F或G。
其中W代表卤素，PG是保护基团，例如-Boc(叔丁氧羰基)、苄基、PMB，R1、R3、R4、R5、R6、R7、X、R2a和R2b如式(I)中所述定义。
方案二：
1)将H和I进行取代反应得到中间体J；
2)中间体J进行卤代反应中间体K；
3)中间体K与相应的胺进行取代得到中间体L或M；
4)中间体L或M脱去保护后与相应的羧酸缩合得到产物N、O或P。
其中W代表卤素，PG是保护基团，例如Boc(叔丁氧羰基)、苄基、PMB，R1、R3、L、R4、R5、R6、R7、X、R2a、R2b和R2e如式(I)中所述定义。
药物组合物
本发明还提供了一种药物组合物，其包含如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，以及任选药用辅料。优选地，所述药用辅料优选药学可接受的载体、稀释剂、赋形剂或其组合。
本发明还提供了一种治疗和/或预防与WRN生物学活性相关的疾病的药物组合物，所述药物组合物包括治疗和/或预防有效量的如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，以及任选药用辅料。优选地，所述药用辅剂优选药学可接受的载体、稀释剂、赋形剂或其组合。
在一些实施方案中，所述的药物组合物还可以包含其他用于治疗和/或预防与WRN生物学活性相关的疾病的药物。
在一些实施方案中，所述与WRN生物学活性相关的疾病为肿瘤或癌症。
在一些实施方案中，所述与WRN生物学活性相关的疾病为MSI肿瘤或MSI癌症。
制备各种含有一定量的活性成分的药物组合物的方法是已知的，或根据本发明的公开内容对于本领域技术人员是显而易见的。如REMINGTON’S PHARMACEUTICAL SCIENCES,Martin,E.W.,ed.,Mack Publishing Company,19th ed.(1995)所述，制备所述药物组合物的方法包括掺入适当的药学赋形剂、载体、稀释剂等。
本发明还提供了一种WRN抑制剂，所述WRN抑制剂包括治疗和/或预防有效量的如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐。
医药用途：
本发明还提供了如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如前所述的药物组合物，在制备WRN抑制剂中的应用。
本发明还提供了如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如前所述的药物组合物，在制备治疗和/或预防与WRN生物学活性相关的疾病的药物中的应用。
本发明还提供了如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如前所述的药物组合物，在治疗和/或预防与WRN生物学活性相关的疾病中的应用。
本发明还提供了如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如前所述的药物组合物，其用于治疗和/或预防与WRN生物学活性相关的疾病。
在一些实施方案中，上述与WRN生物学活性相关的疾病为肿瘤或癌症。
在一些实施方案中，上述与WRN生物学活性相关的疾病为MSI肿瘤或MSI癌症。
本发明还提供了一种治疗和/或预防与WRN生物学活性相关的疾病的方法，其包括给予有需要的个体治疗和/或预防有效量的如前所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如前所述的药物组合物。
在一些实施方案中，上述与WRN生物学活性相关的疾病为肿瘤或癌症。
在一些实施方案中，上述与WRN生物学活性相关的疾病为MSI肿瘤或MSI癌症。
在本发明中，“治疗”一般是指获得需要的药理和/或生理效应。该效应根据完全或部分地预防疾病或其症状，可以是预防性的；和/或根据部分或完全稳定或治愈疾病和/或由于疾病产生的副作用，可以是治疗性的。本文使用的“治疗”涵盖了对患者疾病的任何治疗，包括：(a)预防易感染疾病或症状但还没诊断出患病的患者所发生的疾病或症状；(b)抑制疾病的症状，即阻止其发展；或(c)缓解疾病的症状，即，导致疾病或症状退化。
在本发明中，“受试者”指脊椎动物。在某些实施方案中，脊椎动物指哺乳动物。哺乳动物包括，但不限于，牲畜(诸如牛)、宠物(诸如猫、犬、和马)、灵长类动物、小鼠和大鼠。在某些实施方案中，哺乳动物指人。
在本发明中，“有效量”指在必需的剂量和时间上有效实现期望的治疗或预防效果的量。本发明的物质/分子的“治疗有效量”可根据诸如个体的疾病状态、年龄、性别和体重及该物质/分子在个体中引发期望应答的能力等因素而变化。治疗有 效量还涵盖该物质/分子的治疗有益效果胜过任何有毒或有害后果的量。“预防有效量”指在必需的剂量和时间上有效实现期望的预防效果的量。通常而非必然，由于预防剂量是在疾病发作之前或在疾病的早期用于受试者的，因此预防有效量会低于治疗有效量。在癌症的情况中，药物的治疗有效量可减少癌细胞数；缩小肿瘤体积；抑制(即一定程度的减缓，优选停止)癌细胞浸润到周围器官中；抑制(即一定程度的减缓，优选停止)肿瘤转移；一定程度的抑制肿瘤生长；和/或一定程度的减轻与癌症有关的一种或多种症状。
术语定义：
根据本领域的惯例，在本文结构式中用于描述为该部分或取代基与母核或主结构的连接点的键。
不是出现在两个字母或符号间的破折号“-”用于指示取代基的连接点。例如，C3-6环烷基-(C1-6烷基)r-意为通过(C1-C6烷基)r-连接到分子的其余部分。
本文所用术语“被取代的”是指指定原子或基团上的任一或多个氢被指定基团的选择替代，条件为不超过指定原子的正常价态。
在本说明书的各部分，本发明公开化合物的取代基按照基团种类或范围公开。特别指出，本发明包括这些基团种类和范围的各个成员的每一个独立的次级组合。例如，术语“C1-6烷基”特别指独立公开的甲基、乙基、C3烷基、C4烷基、C5烷基和C6烷基，或者独立公开的“C1-4烷基”，或者独立公开的“C1-3烷基”。
术语“烷基”是指包括具有指定碳原子数的支链和直链饱和脂肪族烃基。例如，“C1-6烷基”是指C1、C2、C3、C4、C5和C6。另外，例如“C1-6烷基”是指具有1至6个碳原子的烷基。烷基可以是未被取代的或被取代的，从而使它的一或多个氢被另一化学基团替代。烷基的实例包括但不限于甲基、乙基、丙基(例如正丙基和异丙基)、丁基(例如，正丁基、异丁基、叔丁基)、戊基(例如正戊基、异戊基、新戊基)等。本领域技术人员应当理解，此处C1-6烷基包含一价的C1-6烷基、二价的C1-6亚烷基，例如C3-C6环烷基-(C1-C6烷基)r-中的C1-C6烷基是指C1-C6亚烷基。
术语“烷氧基”指的是任意上述烷基(例如C1-6烷基、C1-4烷基、C1-3烷基等)，其通过氧原子(-O-)连接到分子的其余部分。
术语“C1-6卤代烷基”或“C1-6卤代烷氧基”，是指所述烷基或烷氧基中一个或多个(如2个、3个)氢原子被卤素原子，如氟、氯、溴取代。所述烷基或烷氧基的定义如上所述。在一些实施方案中，术语“卤代C1-6烷基”优选氟代，例如可以为-CF3、-CHF2、-CH2F、-CH2CH2F、-CH2CHF2、-CH2CF3等。在一些实施方案中，术语“卤代C1-6烷氧基”优选氟代，例如可以为-OCF3、-OCHF2、-OCH2F、-OCH2CH2F、-OCH2CHF2、-OCH2CF3等。
术语“羟基取代的C1-6烷基”是指所述烷基中的一个氢原子被羟基所取代，所 述烷基的定义如上所述。作为示例，所述“羟基取代的C1-6烷基”可以为羟甲基。
术语“烯基”是指包括直链或支链构型并具有一或多个可存在于沿该链的任一稳定点处的碳-碳双键的烃基。例如，“C2-6烯基”是指包括C2、C3、C4、C5和C6。烯基的实例包括但不限于乙烯基、1-丙烯基、2-丙烯基、2-丁烯基、3-丁烯基、2-戊烯基、3-戊烯基、4-戊烯基、2-己烯基、3-己烯基、4-己烯基、5-己烯基、2-甲基-2-丙烯基、4-甲基-3-戊烯基等。
术语“炔基”是指包括直链或支链构型并具有一或多个可存在于沿该链的任一稳定点处的碳-碳三键的烃基。例如“C2-6炔基”是指包括C2、C3、C4、C5和C6炔基。炔基的实例包括但不限于乙炔基、丙炔基、丁炔基、戊炔基、己炔基等。
术语“环烷基”是指环化的烷基，包括单环、双环或多环体系。当环烷基为双环或多环时，其每一个环均应当为饱和的碳环或碳环残基，二环或多环的环烷基每两个环可以的连接方式包括桥接、稠合或螺接。如，C3-10环烷基是指包括C3、C4、C5、C6、C7、C8、C9和C10环烷基。环烷基的实例包括但不限于环丙基、环丁基、环戊基、环己基、等。
术语“环烯基”是指如上所述定义的环烷基中，至少有一个碳碳双键的碳环基，例如
术语“碳环”或“碳环残基”是指任何稳定的3元、4元、5元、6元或7元单环或5元、6元、7元、8元、9元、10元、11元、12元、13元、14元二环或多环，其中任一个环可为饱和、部分饱和、不饱和或芳香族。二环或多环的碳环每两个环可以的连接方式包括桥接、稠合或螺接。这些碳环的实例包括但不限于环丙基、环丁基、环丁烯基、环戊基、环戊烯基、环己基、环己烯基、环庚烯基、环庚基、金刚烷基、环辛基、苯基、萘基、[2,2,2]二环辛烷、等。
术语“芳基”是指环部分中具有5至14个碳原子的单环、二环或三环芳香族烃基，当“芳基”为二环或三环时，其每一个环均为芳香环。二环或三环的芳基每两个环可以的连接方式包括桥接、稠合、螺接。例如苯基和萘基，他们每一个均可被取代。
术语“杂环”、“杂环的”或“杂环基”可互换使用并且是指被取代的和未被取代的4-8元单环或二环基团、8-10元二环或三环基团和10-14元三环或多环基团，其中至少一个环具有至少一个杂原子(O、S或N)，该含杂原子环优选具有1个、2个或3个选自O、S和N的杂原子。该基团中的每一含有杂原子的环皆可含有1或2个氧或硫原子和/或1至4个氮原子，限制条件为每一环中的杂原子总数是4或更小，并且进一步的限制条件为该环含有至少一个碳原子。在一些优选方案中，所述杂原子仅指N或O，且其总数不超过3个，优选仅含1-2个杂原子。 碳和硫原子可任选被氧化，氮原子可任选被季铵化，当化合价允许时，杂环上的环原子可以任选被＝O(氧代)取代。(例如：)。完成二环和三环基团的稠环可仅含有碳原子且可为饱和、部分饱和或完全不饱和，芳香或非芳香的。杂环基团可在任何可用氮或碳原子上连接。在一些优选实施方案中，本文中的杂环基，完成二环和三环基团的稠环为非芳香的环。如前所述，杂环基包括如下所述的“杂环烷基”和“杂环烯基”。示例性杂环基包括但不限于氮杂环丁基、氧杂环丁基、吡咯烷基、咪唑啉基、噁唑烷基、异噁唑啉基、噻唑烷基、四氢呋喃基、哌啶基、哌嗪基、2-氧代哌嗪基、2-氧代哌啶基、2-氧代吡咯烷基、2-氧代氮杂环庚三烯基、1-吡啶酮基、4-哌啶酮基、四氢吡喃基、吗啉基、1,3-二氧杂环戊烷基、奎宁环基、等。
术语“杂环烷基”是指如上所述定义的杂环中，所有环原子均为完全饱和状态的杂环基，例如当为二环或三环时，“杂环烷基”包括“杂桥环烷基”、“杂螺环烷基”、“杂并环烷基”。
术语“杂环烯基”是指如上所述定义的杂环中，至少有一个碳碳双键的杂环基，例如当为二环或三环时，“杂环烯基”包括“杂桥环烯基”、“杂螺环烯基”、“杂并环烯基”。当杂环烯基为二环或三环时，环整体为不饱和且不能形成芳香性。
术语“杂芳基”是指在至少一个环中具有至少一个杂原子(O、N或S)的被取代的和未被取代的上述芳基，包括芳香族5-8元单环基团、8-10元二环基团和10-14元三环基团，该含杂原子环优选具有1个、2个或3个选自O、N或S的杂环原子。杂芳基的每一含有杂原子的环皆可含有1或2个氧或硫原子和/或1至4个氮原子，限制条件为每一环中的杂原子总数是4或更少并且每一环具有至少一个碳原子。为二环或三环的杂芳基每一个环都是芳香族环。
术语“螺环基”是指具有一个共用环原子的双环结构，且每个单环均是具有3-7个碳原子的饱和或不饱和、芳香或非芳香的碳环。示例性的螺环基包括但不限于：螺环[4.5]癸烷、螺环[3.4]辛烷、螺环[2.3]己烷、等。本文中，所述螺环基不包括如上所述定义的芳基。
术语“螺杂环基”是指具有一个共用环原子的双环结构，且每个单环是具有3-8个环原子的饱和或不饱和单环基团，其中至少一个环上具有1或2个环原子是选自N、O或S(O)n的杂原子，其中n是0至2的整数，剩余的环原子为C。另外，杂环基环中的1或2个环碳原子可以任选地被-CO-基团替换。示例性的螺杂环基包括但不限于：5-氮杂螺[2.3]己烷、6-噁螺[3.4]-7-辛酮。本文中，所述螺杂环基不包括如上所述定义的杂芳基。
术语“杂原子”应包括氧、硫和氮。
术语“卤素”应包括“F、Cl、Br、I”。
当术语“不饱和”在文中用于指环或基团时，该环或基团可为完全不饱和或部分不饱和。
当术语“饱和”在文中用于指环或基团时，如无特殊说明，该环或基团应当为完全饱和。
从所有上述描述中，对本领域技术人员显而易见的是，其名称是复合名称的任意基团，例如“吡啶并噻吩基”，从其衍生的部分，从吡啶基或噻吩基来构建，优选从噻吩基来构建；其他类似的复合名称可以参照前述内容进行理解。
术语“任选”表示可以选，也可以不选。例如，“任选被1至3个Rd取代的C1- 6烷基”，其表示，所述C1-6烷基可以被1至3个Rd所取代，也可以不被1至3个Rd所取代。其他类似的定义可以参照前述内容进行理解。
在整个说明书中，基团和其取代基可由本领域技术人员选择以提供稳定部分和化合物和可用作医药上可接受的化合物的化合物和/或可用于制备医药上可接受的化合物的中间体化合物。
本文中，除非以其他方式明确指出，在本文中通篇采用的描述方式“…各自独立地选自”既可以是指在不同基团中，相同或不同的符号之间所表达的具体选项之间互相不影响，也可以表示在相同的基团中，相同或不同的符号之间所表达的具体选项之间互相不影响。
术语“XXX被一个或多个选自YYY的取代基取代”指的是，XXX可以被一个或多个取代基在任意可被取代的位置进行取代，所述取代基选自YYY。当XXX被多个选自YYY的取代基在任意可取代的位置进行取代时，多个取代基可以相同，也可以不同。其中，多个为2个或更多，优选2个、3个或4个，更优选2个或3个。例如，C1-6烷基被一个或多个选自氰基和羟基的取代基取代在任意可取代的位置，其表示，C1-6烷基可以被一个或多个氰基在任意可取代的位置取代，也可以被一个或多个羟基在任意可取代的位置取代，还可以被一个或多个氰基和羟基(例如一个氰基和一个羟基，或者两个氰基和一个羟基，或者两个氰基和两个羟基等)在任意可取代的位置同时取代。
提及“XXX中的一个或多个H原子可进一步地被一个或多个相同或不同的YYY取代”时是指XXX中的H原子可以被YYY取代，也可以不被取代；XXX中的H原子可以被一个或多个相同的YYY取代，也可以被一个或多个不同的YYY取代，每个取代基之间不应符号相同而相互干涉。
发明效果：
本发明的式(Ⅰ)所示的化合物显示具有良好的WRN抑制作用，能够作为与该作用有关的疾病的治疗和/或预防有关的药物，尤其可用于MSI肿瘤相关疾病的治疗和/或预防。
具体实施方式
应该理解，此处采用的术语目的在于描述具体的实施方案，并非意在限制。此外，尽管类似或者等价于此处描述的任何方法、装置和材料均可用于实施或者测试本发明，但是现在描述的是优选的方法、装置和材料。
化合物的结构是通过核磁共振(NMR)或质谱(MS)来确定的。NMR的测定是用Bruker ASCENA-400核磁仪，测定溶剂为氘代二甲基亚砜(DMSO-d6)、氘代氯仿(CDCl3)、氘代甲醇(CD3OD)，内标为四甲基甲硅烷(TMS)，化学位移是以10- 6(ppm)作为单位给出。
反应监控及MS的测定使用Thermofisher ESQ(ESI)质谱仪。
HPLC的测定使用赛默飞U3000DAD高压液相色谱仪(GL Sciences ODS-HL HP 3μm 3.0*100mm色谱柱)。
薄层层析硅胶板使用青岛海洋GF254硅胶板，薄层色谱法(TLC)使用的硅胶板采用的规格是0.15～0.2mm，薄层层析分离纯化产品采用的规格是0.9～1.0mm的高效薄层色谱制备板。柱层析使用青岛海洋200～300目硅胶为载体，展开剂所使用的体系有A：二氯甲烷和甲醇体系；B：石油醚和乙酸乙酯体系，溶剂的体积比根据化合物的极性不同而进行调节。中压制备液相纯化时使用的是biotage isera one型制备液相。制备型液相色谱仪(prep-HPLC)型号为Agilent 1290Infinity 2代。
以下实施例中，如无特别说明，所有反应原料可以从萨恩化学技术(上海)有限公司、上海韶远试剂有限公司、南京药石科技股份有限公司、江苏艾康生物医药研发有限公司、上海毕得医药科技有限公司等厂家购买获得。
简写说明
中间体int-1的制备：4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6- 基)哌嗪-1-甲酸叔丁酯
步骤1:将3-氧代戊酸甲酯(int-1a，70g，538.05mmol)溶于DCM(500mL)中，加入NBS(100.6g,564.95mmol)，然后加入TSOH.H2O(20.4g,107.61mmol)，室温下搅拌3小时。抽滤，滤液经减压浓缩得到粗品2-溴-3-氧代戊酸甲酯(int-1b,126.5g,602.87mmol)。MS Calcd:207.97；MS Found:206.98([M-H]-).
步骤2:将2-溴-3-氧代戊酸甲酯(int-1b，10g，47.85mmol)，哌嗪-1-甲酸叔丁酯(int-1c,49.0g,263.16mmol)，碳酸钾(39.7g,287.08mmol)溶入到MeCN(100mL)，常温下搅拌30分钟。过滤，滤液用稀盐酸中和水洗后，DCM萃取，浓缩有机相，残余物经柱层析得到4-(1-甲氧基-1,3-二氧代戊烷-2-基)哌嗪-1-甲酸叔丁酯(int-1d,10.97g,34.89mmol,72.9％)。MS Calcd:314.18；MS Found:315.23([M+H]+)。
步骤3：室温下，将H3PO4(4.5g,46.01mmol)加入到5-溴-2H-1,2,4-三唑-3-胺(int-1e,5g,30.67mmol)和4-(1-甲氧基-1,3-二氧代戊烷-2-基)哌嗪-1-甲酸叔丁酯(int-1d,10.6g,33.74mmol)的无水乙醇(50mL)溶液中，并加热到80℃搅拌两天。加入饱和碳酸氢钠淬灭，并调pH为6-7，浓缩反应液，乙酸乙酯萃取，无水硫酸钠干燥有机相，浓缩快速柱层析(DCM＝95％，MeOH＝5％)得到4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-1,2.09g,4.89mmol,15.9％)。MS Calcd:426.10；MS Found:427.07([M+H]+)。
中间体int-2的制备：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺
步骤1：向50mL茄形瓶中加入4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-1,400mg,0.94mmol)和溶剂1,4-二氧六环(5mL)。然后向其中加N-(2-氯-4-(三氟甲基)苯基)-2-碘乙酰胺(374mg,1.03mmol)，DIPEA(383mg,2.81mmol)，85℃室温搅拌5小时，向其中加入水(20mL)，用EA萃取(20mL×3)，饱和食盐水洗涤有机相(20mL)，无水硫酸钠干燥，减压旋干得粗品，粗品通过柱层析(PE:EA＝1:2)纯化得到4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌 嗪-1-甲酸叔丁酯(int-2a,430mg,0.65mmol,69.3％yield)。MS Calcd:662.89；MS Found:564.15([M+H-100]+).
步骤2：向50mL茄形瓶中加入4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2a,350mg,0.53mmol)，2-(3,6-二氢-2H-吡喃-4-基)-4,4,5,5-四甲基-1,3,2-二氧杂硼烷(166mg,0.79mmol)和溶剂1,4-二氧六环(3mL)，H2O(1.5mL)。然后向其中加Pd(dppf)Cl2(39mg,0.053mmol)，磷酸钾(365mg,1.58mmol)，氮气保护下，80℃搅拌过夜，向其中加入水(20mL)，用EA萃取(20mL×3)，饱和食盐水洗涤有机相(20mL)，无水硫酸钠干燥，减压旋干得粗品，粗品通过柱层析(PE:EA＝1:2)纯化得4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2b,220mg,0.33mmol,61.8％yield)MS Calcd:666.10；MS Found:610.19([M-56]+).
步骤3：向50mL茄形瓶中加入4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2b,220mg,0.33mmol)和溶剂DCM(5mL)。然后向其中加三氟乙酸(1mL)，室温搅拌过夜，LCMS检测原料反应完毕，向其中加入饱和碳酸氢钠溶液(20mL)，用EA萃取(20mL×3)，饱和食盐水洗涤有机相(20mL)，无水硫酸钠干燥，减压旋干得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,140mg,0.25mmol,75.7％yield)。MS Calcd:565.18；MS Found:566.26([M+H]+).
中间体int-3的制备：2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺
步骤1：将5-溴-2H-1,2,4-三唑-3-氨(int-1e,2.0g，12.3mmol)和3-氧代戊酸甲酯(int-1a，2.2g,15.3mmol)加入到25mL圆底烧瓶中，加入AcOH(7mL)，加热到80℃搅拌过夜。冷却到室温后，在0℃下搅拌1h，过滤，滤饼用少量EtOH(10mL) 洗涤，烘干；滤液浓缩至干，少量EtOH洗涤(10mL)，固体烘干后得第二部分产物，合并得2-溴-5-乙基-[1,2,4]三唑并[1,5-a]嘧啶-7(4H)-酮(int-3a,1.63g,6.71mmol,54.5％yield)。MS Calcd:241.98、243.98；MS Found:242.98、244.98([M+H]+).
步骤2：将2-溴-5-乙基-[1,2,4]三唑并[1,5-a]嘧啶-7(4H)-酮(int-3a,1.63g，6.71mmol)和N-[2-氯-4-(三氟甲基)苯基]-2-碘代乙酰胺(2.67g,7.35mmol)加入到25mL圆底烧瓶中，加入1,4-二氧六环(7mL)，氮气保护下于80℃搅拌4h。降到室温，析出白色沉淀，过滤，滤液浓缩，加EA后再次析出白色固体，过滤，EA洗涤(10mL)，合并固体，减压除去溶剂得2-(2-溴-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)-N-[2-氯-4-(三氟甲基)苯基]乙酰胺(int-3b,3.02g,6.31mmol,94.0％yield)。MS Calcd:476.98、478.98；MS Found:477.94、479.95([M+H]+).
步骤3：将2-(2-溴-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)-N-[2-氯-4-(三氟甲基)苯基]乙酰胺(int-3b,3.0g,6.27mmol)和3,6-二氢-2H-吡喃-4-硼酸片呐醇酯(1.71g，8.14mmol)加入到100mL两口瓶中，依次加入磷酸钾(3.99g，18.80mmol)、Pd(dppf)Cl2(458mg,0.63mmol)、1,4-二氧六环(40mL)和纯化水(20ml)，体系呈棕色悬浊液，氮气保护下于80℃搅拌2h。降到室温，旋去多余溶剂，加入DCM(100mL)和水(100mL)，分液，水相用DCM萃取(50mL*3)，合并有机相，饱和食盐水洗涤(100mL)，减压除去溶剂，加入EA/PE(2:1)出现棕色沉淀，柱层析(DCM:MeOH＝10:1)得N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-3c,1.58g,3.28mmol,52.3％yield)。MS Calcd:481.11；MS Found:479.94([M-H]-).
步骤4：将N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-3c,1.35g，2.80mmol)加入到50mL烧瓶中，依次加入NBS(548mg)和乙腈(50mL)，加热到80℃继续反应6h。冷却后析出固体，过滤，滤饼，滤饼经柱层析(MeOH:DCM＝1:10)纯化得标题化合物：2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(int-3,0.51g,0.91mmol,32.5％yield)。MS Calcd:559.02、561.02；MS Found:559.8、562.0([M+H]+).
中间体int-4的制备：N-(3-氟-2-甲基-4-(三氟甲基)苯基)-2-碘乙酰胺
步骤1：向250mL茄形瓶中加入4-溴-2-氟-1-(三氟甲基)苯(int-4a,9.5g,39.09 mmol)和溶剂THF(20mL)并降温至-78℃。氮气保护下向其中缓慢滴加LDA(4.6g,43.00mmol)，在-78℃搅拌2小时，然后向其中加入碘甲烷(4.76g,76mL,58.64mmol)，升温至室温并反应过夜。在冰浴下加入饱和氯化铵淬灭，用二氯甲烷萃取(50mL*3)，有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得到粗品：1-溴-3-氟-2-甲基-4-(三氟甲基)苯(int-4b,9.0g,35.02mmol,89.6％yield).1H NMR(400MHz,DMSO-d6)δ7.70(d,J＝8.4Hz,1H),7.58(t,J＝8.4Hz,1H),2.35(d,J＝2.8Hz,3H).
步骤2：向250mL茄形瓶中加入1-溴-3-氟-2-甲基-4-(三氟甲基)苯(int-4b,9.0g,35.02mmol)，氨基甲酸叔丁酯(4.5g,38.52mmol)和溶剂1,4-二氧六环(20mL)，向其中加入Xantphos(1.4g,2.45mmol)，Pd2(dba)3(1.6g,1.75mmol)，Cs2CO3(17.1g,52.53mmol)，氮气保护下升温至85℃反应3小时。冷却至室温，过滤，滤饼用二氯甲烷洗涤，有机相减压旋干得到粗品，粗品通过快速柱层析(PE:EA＝20:1)得到：3-氟-2-甲基-4-(三氟甲基)苯基)氨基甲酸叔丁酯(int-4c,7.3g,24.89mmol,71.1％yield).MS Calcd:293.10；MS Found:292.1([M-H]-).
步骤3：向250mL茄形瓶中加入3-氟-2-甲基-4-(三氟甲基)苯基)氨基甲酸叔丁酯(int-4c,7.26g,24.75mmol)，盐酸甲醇溶液(20mL)，室温下反应两小时。向反应液中加入氢氧化钠调至pH至中性，过滤除去不溶物，有机相用2*30mL水洗涤，无水硫酸钠干燥有机相，浓缩得到粗品：3-氟-2-甲基-4-三氟甲基苯胺(int-4d,4.3g,22.27mmol,89.6％yield).MS Calcd:193.05；MS Found:192.11([M-H]-).
步骤4：向100mL茄形瓶中加入3-氟-2-甲基-4-三氟甲基苯胺(int-4d,4.3g,22.27mmol)，二氯甲烷(30mL)，三乙胺(6.17mL,44.54mmol)，冰浴下滴加氯乙酰氯(2.65mL,26.72mmol)的二氯甲烷溶液(10mL)，然后自然恢复室温反应过夜。反应液减压旋干得到粗品，粗品通过快速柱层析(PE:EA＝3:1)得到：2-氯-N-[3-氟-2-甲基-4-三氟甲基苯基]乙酰胺(int-4e,5.5g,18.77mmol,84.3％yield).MS Calcd:269.02；MS Found:268.07([M-H]-).
步骤5：向250mL茄形瓶中加入2-氯-N-[3-氟-2-甲基-4-三氟甲基苯基]乙酰胺(int-4e,5.5g,18.77mmol)，碘化钾(6.8g,40.80mmol)和溶剂丙酮(30mL)，然后50℃反应3小时。反应液减压旋干得到粗品，粗品通过快速柱层析(PE:EA＝5:1)得到：N-[3-氟-2-甲基-4-三氟甲基苯基]-2-碘乙酰胺((int-4,6.12g,16.95mmol,82.7％yield).MS Calcd:360.96；MS Found:359.99([M-H]-).
中间体int-5的制备：4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯
步骤1：把4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-1,14g,32.8mmol)在二氧六环(150mL)和水(50mL)的250mL的三口瓶中，之后依次加入3,6-二氢-2H-吡喃-4-硼酸片呐醇酯(10.3g,49.1mmol)，K2CO3(9.1g,65.6mmol)，Pd(dppf)cl2(1.2g,1.64mmol)，在氮气的氛围下置换三次，加热到100℃反应14小时，冷却到室温，加入300mL水，用300mL二氯甲烷萃取3次，有机相用饱和食盐水洗涤，无水硫酸钠干燥，旋干，参与物经柱层析(DCM/MeOH＝100/0-95/5)洗涤获得粗产品，粗产品用甲基叔丁基醚打浆获得纯的4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-5,9.2g,65％yield)。MS Calcd.:430.2；MS Found:431.2[M+H]+.1H NMR(400MHz,DMSO-d6)δ13.00(s,br,1H),6.81(br,1H),4.28-4.27(m,2H),3.93-3.90(m,2H),3.83-3.80(m,2H),3.37-3.33(m,2H),2.90-2.89(m,2H),2.79-2.73(m,2H),2.64-2.60(m,2H),2.51-2.50(m,2H),1.42(s,9H),1.19(t,J＝7.2Hz,3H).
中间体int-6的制备：(R)-4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯
步骤1：向2L三口瓶中将3-氧代戊酸乙酯(int-6a,100.00g,693.63mmol)溶解于二氯甲烷(1L)中，依次加入NBS(129.62g,728.31mmol)，TsOH*H2O(26.39g,138.73mmol)。得到的反应混合物于室温下搅拌过夜。反应完毕后将反应混合物倒入饱和食盐水水中(1L)，用二氯甲烷萃取两遍(1L*2)。合并的有机相，用盐水反洗两次，经无水硫酸钠干燥，过滤，旋干。得到的粗品，经快速硅胶柱纯化(石油醚/乙酸乙酯＝10：1)得到2-溴-3-氧代戊酸乙酯(int-6b,120.0g，收率为77.6％)。MS Calcd.:221.99；MS Found:229.2[M+H]+.
步骤2：向1L单口瓶中将2-溴-3-氧代戊酸乙酯(int-6b,60.00g,268.98mmol)溶于乙腈中(300mL)，依次加入K2CO3(74.36g,537.96mmol)and(R)-1-N-Boc-2-甲基哌嗪(56.56g,282.43mmol)。得到的反应混合物于室温下搅拌过夜。滤除掉未溶的无机盐，将滤液浓缩至干。所得残留物经快速硅胶柱纯化(石油醚/乙酸乙酯＝5：1)得(2R)-4-(1-乙氧基-1,3-二氧代戊烷-2-基)-2-甲基哌嗪-1-甲酸叔丁酯(int-6c,64.0g,收率为69.5％)。MS Calcd.:342.22；MS Found:343.22[M+H]+。
步骤3：称取(2R)-4-(1-乙氧基-1,3-二氧代戊烷-2-基)-2-甲基哌嗪-1-甲酸叔丁酯(int-6c,91.32g，266.68mmol)置于1L单口瓶中，加入乙醇(150mL)溶解，再依次加入5-溴-2H-1,2,4-三唑-3-氨(int-1e,47.81g,293.35mmol)和磷酸(27.44g，280.01mmol)，整个反应体系转换氮气三次，于90℃下反应搅拌44小时后，冷 却至室温。向单口瓶中加入DIPEA(103.40g,800.04mmol)，Boc2O(29.10g,133.34mmol)，室温反应2h。反应完毕后，将反应混合物倒入饱和食盐水水中(500mL)，用乙酸乙酯萃取三遍(1L*3)。合并的有机相，用盐水反洗两次，经无水硫酸钠干燥，过滤，旋干，得到残留物。所得残留物经快速硅胶柱纯化(二氯甲烷/甲醇＝17：3)得标题化合物：(R)-4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(int-6,50.0g，收率为38.6％)。MS Calcd.:442.12；MS Found:443.2[M+H]+。1H NMR(400MHz,DMSO-d6)δ4.16(d,J＝23.6Hz,1H),3.81-3.69(m,1H)，3.58-3.49(m,1H)，3.41-3.31(m,1H)，3.19-3.09(m,1H)，3.09-2.89(m,1H)，2.88-2.77(m,1H)，2.66-2.54(m,2H)，1.42(s,9H)，1.29-1.26(m,3H)，1.18(t,J＝7.6Hz,3H).
中间体int-7的制备：4-羟基异噁唑-3-甲酸
步骤1：将4-氯-3-氧代丁酸乙酯(int-7a,10g,60.75mmol)溶于AcOH(48mL)溶液中，置于0℃条件下缓慢加入NaNO2(5.2g,75.94mmol)的H2O(40mL)溶液，反应体系变为红色，然后升至室温搅拌过夜。向反应液中加入100mL H2O，乙酸乙酯萃取100mL*3次，饱和食盐水(100mL)洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析分离纯化(PE:EA＝3:1)得4-氯-2-(羟基亚胺)-3-氧代丁酸乙酯(int-7b,11g,56.82mmol,93.5％yield)。
步骤2：将4-氯-2-(羟基亚胺)-3-氧代丁酸乙酯(int-7b,12g,61.98mmol)和尿素(29.8g,495.87mmol)依次加入DMF(50mL)溶液中，升温至100℃搅拌20min。向反应液中加入150mL水，DCM萃取(100mL*3)，饱和食盐水洗涤四次，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析分离纯化(PE:EA＝2:1)得4-羟基异噁唑-3-甲酸乙酯(int-7c,8g,50.92mmol,82.2％yield)。MS Calcd:157.04；MS Found:158.08([M+H]+).
步骤3：将4-羟基异噁唑-3-甲酸乙酯(int-7c,2000mg,12.73mmol)溶于THF(20mL)中，冰浴下加入NaOH(2546mg,63.65mmol)，室温反应2.0h。将溶剂旋干后用柱层析分离(DCM:MeOH＝1:1)得标题化合物：4-羟基异噁唑-3-甲酸(int-7,1000mg,7.75mmol,60.8％yield)。MS Calcd:129.01；MS Found:128.0([M+H]+).
中间体int-8的制备：2-(4-甲氧基环己-1-烯-1-基)-4,4,5,5-四甲基-1,3,2-二氧杂硼烷
步骤1：将4-甲氧基环己-1-酮(int-8a,1g,7.8mmol),1,1,1-三氟-N-苯基-N-(三 氟甲基)磺酰基)甲磺酰胺(2.8g,7.8mmol)加入到干燥的THF中(20mL)，降低温度到-78℃，然后滴加LiHMDS的THF溶液(1N，7.8mL)，在此温度下搅拌2小时，然后缓慢恢复到室温，并搅拌过夜，用饱和氯化铵水溶液淬灭，并用EA萃取(20mLx2)，有机层经合并，无水硫酸钠干燥，抽滤，浓缩，剩余物经柱层析纯化(EA：PE＝1:5)，得到粗品4-甲氧基环己-1-烯-1-基三氟甲磺酸酯(int-8b,1.3g,5mmol,64％)。
步骤2：将4-甲氧基环己-1-烯-1-基三氟甲磺酸酯(int-8b,1.3g,5mmol)，4,4,4',4',5,5',5',5'-八甲基-2,2'-双(1,3,2-二氧杂硼烷)(1.9g,7.5mmol)，醋酸钾(1.5g,15mmol),1,1-双(二苯基膦)二荗铁二氯化钯(366mg,0.5mmol)加入到二氧六环(15mL)中，氮气置换3分钟，升高温度到85℃，搅拌3小时，减压浓缩，剩余物倒入水中，用乙酸乙酯萃取(20mLx3)，合并有机层，经减压浓缩，剩余物经柱层析纯化(EA：PE＝1:20)得到标题化合物：2-(4-甲氧基环己-1-烯-1-基)-4,4,5,5-四甲基-1,3,2-二氧杂硼烷(int-8,910mg,3.82mmol，76％).MS Calcd:238.17；MS Found:239.21([M+H]+)。
中间体1-c的制备：2-(2-(3,6-二氢吡喃-4-基)-5-乙基-7-氧代-6-哌嗪-1-基-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)基)-N-(2-甲基-4-三氟甲基苯基)乙酰胺
步骤1：在100mL单口瓶中依次加入4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-1,1000mg,2.34mmol)，2-碘-N-(2-甲基-4-三氟甲基苯基)乙酰胺(880mg,2.57mmol)，DIPEA(907mg,7.02mmol)和溶剂DMF(5mL)，室温搅拌过夜。LCMS监控反应完全，反应液加入100mL DCM，3*30mL水洗涤，有机相无水硫酸钠干燥，过滤，粗品快速柱层析纯化(MeOH:DCM＝0:100％to 10％:90％in 30min)得4-(2-溴-5-乙基-4-(2-(2-甲基-4-(三氟甲基苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(1-a,1200mg,1.87mmol,79％yield).MS Calcd:641.16；MS Found:640.05([M-H]-).
步骤2：在30mL微波管中加入反应物4-(2-溴-5-乙基-4-(2-(2-甲基-4-(三氟 甲基苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(1-a,900mg,1.40mmol)，2-(3,6-二氢吡喃-4-基)-4,4,5,5-四甲基-1,3,2-二氧杂硼烷(441mg,2.10mmol)，Pd(dppf)Cl2(102mg,0.14mmol)，无水磷酸钾(967mg,4.20mmol)和溶剂1,4-二氧六环(10mL)以及水(2mL)，氮气置换3次，90℃微波搅拌2hr。LCMS监控反应完全，反应液硅藻土过滤，3*10mL甲醇洗涤滤饼，合并滤液，减压旋干。粗品快速柱层析纯化(MeOH:DCM＝0:100％to 10％:90％in 30min)得4-(2-(3,6-二氢吡喃-4-基)-5-乙基-4-(2-(2-甲基-4-三氟甲基苯基氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(1-b,820mg,1.27mmol,90％yield),MS Calcd:645.29；MS Found:644.23([M-H]-).
步骤3：在100mL单口瓶中加入4-(2-(3,6-二氢吡喃-4-基)-5-乙基-4-(2-(2-甲基-4-三氟甲基苯基氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(1-b,500mg,0.77mmol)，盐酸二氧六环溶液(4M,5mL)和溶剂DCM(10mL)，室温搅拌2hr，溶液变浑浊。反应液减压旋干得粗品2-(2-(3,6-二氢吡喃-4-基)-5-乙基-7-氧代-6-哌嗪-1-基-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)基)-N-(2-甲基-4-三氟甲基苯基)乙酰胺(1-c,380mg,0.7mmol,89％yield)。MS Calcd:545.24；MS Found:546.21([M+H]+).
实施例1 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物3)的制备
步骤1：将乙氧基亚甲基丙二酸二乙酯(3-a,1.5g,6.94mmol)、甲基肼硫酸盐(1.0g，6.94mmol)分别加入到10mL水溶液中，于100℃条件下搅拌6小时。将反应液冷却至室温，析出固体，抽滤得粗品5-羟基-1-甲基-1H-吡唑-4-甲酸乙酯(3-b,350mg,2.06mmol,29.6％yield)。MS Calcd:170.17；MS Found:171.11([M+H]+).
步骤2：将5-羟基-1-甲基-1H-吡唑-4-甲酸乙酯(3-b,250mg,1.47mmol)溶于6mL甲醇/水(5：1)混合溶液中，加入氢氧化钾(824mg,14.7mmol)，升温至常温回流搅拌过夜，TLC监测反应完成，将反应液完全浓缩，向残余物中缓慢加入5N HCl溶液，直至固体析出，抽滤得5-羟基-1-甲基-1H-吡唑-4-甲酸(3-c,150mg,1.06mmol,71.9％yield)。MS Calcd:142.11；MS Found:143.05([M+H]+).
步骤3：将5-羟基-1-甲基-1H-吡唑-4-甲酸(3-c,78mg,0.55mmol)和1-氯- N,N,2-三甲基丙烯胺(80mg,0.60mmol)分别加入到5mL二氯甲烷溶液中，室温下搅拌3小时，然后加入2-(2-(3,6-二氢吡喃-4-基)-5-乙基-7-氧代-6-哌嗪-1-基-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)基)-N-(2-甲基-4-三氟甲基苯基)乙酰胺(1-c,150mg,0.27mmol)、DIPEA(178mg,1.37mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(3，8mg,0.01mmol,4％yield)。MS Calcd:669.26；MS Found:670.27([M+H]+).1H NMR(400MHz,DMSO)δ10.42(brs,1H),10.02(s,1H),7.88(s,1H),7.72(d,J＝8.4Hz,1H),7.63(s,1H),7.54(d,J＝8.4Hz,1H),6.83(s,1H),5.25(s,2H),4.30-4.16(m,4H),3.81(t,J＝5.2Hz,2H),3.66(s,3H),3.51-3.45(m,2H),3.17-3.01(m,4H),2.72(d,J＝10.8Hz,2H),2.54-2.52(m,2H),2.36(s,3H),1.21(t,J＝7.6Hz,3H).
实施例2 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物8)的制备
步骤1：将5-羟基-1-甲基-1H-吡唑-4-甲酸(3-c,45mg,0.32mmol)和1-氯-N,N,2-三甲基丙烯胺(46mg,0.35mmol)分别加入到5mL二氯甲烷溶液中，室温下搅拌3小时，然后加入N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,90mg,0.16mmol)、DIPEA(102mg,0.8mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(8，10mg,0.01mmol,9.1％yield)。MS Calcd:689.21；MS Found:690.32([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.45(s,1H),8.08(d,J＝8.4Hz,1H),7.99(d,J＝2.0Hz,1H),7.90(s,1H),7.74(dd,J＝8.4,2.0Hz,1H),6.85(t,J＝2.0Hz,1H),5.34(s,2H),4.28-4.20(m,4H),3.82(t,J＝5.6Hz,2H),3.68(s,3H),3.50–3.44(m,2H),3.11–2.99(m,4H),2.74(d,J＝10.8Hz,2H),2.54-2.52(m,2H),1.22(t,J＝7.6Hz,3H).
实施例3 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物14)的制备
步骤1：将2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(int-3,100mg,0.18mmol)，(R)-2-甲基哌嗪-1-甲酸叔丁酯(180mg，0.9mmol)，四氟硼酸银(35mg，0.18mmol)加入到干燥的DMSO(2mL)中，然后氮气保护下，升高温度到120℃，搅拌4小时，原料反应完毕，冷却到室温，加入饱和碳酸氢钠水溶液，然后用乙酸乙酯萃取(10mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，剩余物经柱层析纯化(EA：PE＝1:1)，得到(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(14-a,74mg,0.11mmol，60％)。MS Calcd:679.25；MS Found:680.33([M+H]+)。
步骤2：将(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(14-a,74mg,0.11mmol)加入到DCM中(5mL)，然后加入三氟乙酸(1mL)，在室温下搅拌4小时，原料反应完毕，减压浓缩，剩余物加入饱和碳酸氢钠水溶液，然后用DCM萃取(5mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，得到(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,51mg,0.09mmol,80％)。MS Calcd:579.20；MS Found:580.31([M+H]+)。
步骤3：将5-羟基-1-甲基-1H-吡唑-4-甲酸(3-c,17mg,0.11mmol)加入到DCM中(1mL)，然后加入1-氯-N，N，2-三甲基丙基-1-烯-1-胺(16mg，0.11mmol)，室温下搅拌2小时，然后加入到(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,51mg,0.09mmol)和DIPEA(70mg，0.54mmol)的二氯甲烷溶液中，并在室温下搅拌过夜。然后加入水(3mL)，用DCM萃取(3mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，剩余物经Prep-HPLC纯化，得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶- 4(7H)-基)乙酰胺(14,8mg,0.011mmol,12.6％yield)。MS Calcd:703.22；MS Found:704.31([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.70(s,1H),10.55(s,1H),8.09(d,J＝8.8Hz,1H),7.97(d,J＝2.0Hz,1H),7.90(s,1H),7.73(dd,J＝8.8,2.0Hz,1H),6.85(t,J＝2.0Hz,1H),5.33(s,2H),4.35-4.30(m,1H),4.35-4.26(m,3H),3.82(t,J＝5.6Hz,2H),3.68(s,3H),3.52-3.50(m,2H),3.17-3.12(m,2H),2.98-2.93(m,1H),2.75-2.62(m,4H),1.45-1.35(m,3H),1.24(t,J＝7.2Hz,3H).
实施例4 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物15)的制备
步骤1：将(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,60mg,0.10mmol)，DIPEA(13mg，0.10mmol)，1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a，60mg，0.3mmol)加入到2-甲基-2-丁醇中，油浴升温到180℃，搅拌30分钟，将反应体系溶解到甲醇中，直接prep-HPLC纯化，得标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(15,15mg,0.020mmol,21％yield)。MS Calcd:729.24；MS Found:730.31([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.09(dd,J＝8.8,6.0Hz,1H),7.99(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.60(s,1H),6.97–6.73(m,1H),5.35(d,J＝2.8Hz,2H),4.70-4.50(m,1H),4.28-4.26(m,2H),3.82(t,J＝5.6Hz,2H),3.70(d,J＝11.2Hz,1H),3.54–3.26(m,3H),3.19-3.14(m,1H),2.98-2.93(m,1H),2.79(d,J＝10.0Hz,1H),2.71-2.69(m,1H),2.56-2.46(m,2H),2.36-2.34(m,1H),1.45-1.41(m,2H),1.24(t,J＝7.2Hz,3H),1.02-0.95(m,5H).
实施例5 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-1-甲基-1H-吡唑-4-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物16)的制备
步骤1：将3-羟基-1-甲基-1H-吡唑-4-甲酸乙酯(16-a,100mg,0.59mmol)溶于6mL甲醇/水(5：1)混合溶液中，加入氢氧化钾(200mg,3.54mmol)，升温至回流搅拌过夜，TLC监测反应完成，将反应液完全浓缩，向残余物中缓慢加入5N HCl溶液，直至固体析出，抽滤得产物3-羟基-1-甲基-1H-吡唑-4-甲酸(16-b,70mg,0.49mmol,83.8％yield)。MS Calcd:142.11；MS Found:143.07([M+H]+).
步骤2：将3-羟基-1-甲基-1H-吡唑-4-甲酸(16-b,40mg,0.28mmol)和1-氯-N,N,2-三甲基丙烯胺(42mg,0.31mmol)分别加入到5mL二氯甲烷溶液中，室温下搅拌3小时，然后加入N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-280mg,0.14mmol)、DIPEA(90mg,0.71mmol)，继续升温至40℃搅拌5h，向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩后残余物溶于4mL甲醇/水(5:1)混合溶剂中，然后加入氢氧化钠(15mg,0.37mmol)，置于室温搅拌过夜。LCMS监测反应完成，将反应液调至pH＝2，prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-1-甲基-1H-吡唑-4-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(16，3mg,0.004mmol,3％yield)。MS Calcd:689.21；MS Found:690.31([M+H]+).1H NMR(400MHz,DMSO)δ10.45(s,1H)8.37(s,1H),8.08(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.90(s,1H),7.73(dd,J＝8.8,2.0Hz,1H),6.84(s,2H),5.32(s,2H),4.28-4.23(m,3H),3.82(t,J＝5.2Hz,2H),3.67(s,3H),3.50-3.30(m,6H),3.04-2.98(m,2H),2.74(d,J＝10.0Hz,2H),1.25(t,J＝7.2Hz,3H).
实施例6 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物17)的制备
步骤1：将乙氧基甲叉丙二酸二乙酯(17-a,1.41g,6.53mmol)、1-环丁基肼盐酸盐(0.8g，6.53mmol)和碳酸钾(1.8g,13.05mmol)分别加入到10mL水溶 液中，于100℃条件下搅拌6小时。TLC监测反应完成，将反应液冻干，加入少量甲醇溶解，用5N HCl调至pH＝2，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,900mg,4.28mmol,65.6％yield)。MS Calcd:210.10；MS Found:211.12([M+H]+).
步骤2：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,50mg,0.24mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2 45mg,0.06mmol)加入4mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。LCMS监测反应完成，将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(17,5mg,0.01mmol,3％yield)。MS Calcd:729.24；MS Found:730.24([M+H]+).1H NMR(400MHz,DMSO)δ10.38(s,1H),8.08(d,J＝8.8Hz,1H),7.97(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),7.70(s,1H),6.85(s,1H),5.33(s,2H),4.83-4.76(m,1H),4.36-4.26(m,2H),4.25(s,2H),3.82(t,J＝5.2Hz,2H),3.52-3.17(m,6H),3.00(t,J＝7.2Hz,2H),2.76(d,J＝11.2Hz,2H),2.32–2.23(m,2H),1.83-1.72(m,2H),1.26–1.19(m,5H).
实施例7 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物18)的制备
步骤1：将乙氧基甲叉丙二酸二乙酯(17-a,2.0g,9.22mmol)、环丙基肼盐酸盐(1.0g，9.22mmol)和碳酸钾(2.55g,18.43mmol)分别加入到10mL水溶液中，于100℃条件下搅拌6小时。TLC监测反应完成，将反应液冻干，加入少量甲醇溶解，用5N HCl调至pH＝2，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a,1.2g,6.12mmol,66.4％yield)。MS Calcd:196.08；MS Found:197.12([M+H]+).
步骤2：将1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a,117mg,0.6mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2 45mg,0.06mmol)加入4 mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。LC-MS监测反应完成，将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(18,3mg,0.004mmol,6.2％yield)。MS Calcd:715.22；MS Found:716.28([M+H]+).1H NMR(400MHz,DMSO)δ10.36(s,1H),8.08(d,J＝8.8Hz,1H),7.97(d,J＝2.0Hz,1H),7.72(d,J＝8.8,2.0Hz,1H),7.55(s,1H),6.83(s,1H),5.32(s,2H),4.36-4.26(m,4H),3.84-3.81(m,2H),3.51-3.17(m,5H),3.05-2.98(m,2H),2.75(d,J＝11.2Hz,2H),1.26-1.20(m,5H),0.99-0.93(m,2H).
实施例8 2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物19)的制备
步骤1：将4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-1,2.2g,5.15mmol)、N-(5-氟-2-甲基-4-(三氟甲基)苯基)-2-碘乙酰胺(2.0g，5.67mmol)和DIPEA(2.0g,15.45mmol)分别加入到60mL二氧六环溶液中，于80℃条件下搅拌4小时。TLC监测反应完成，将反应液减压浓缩，残余物经硅胶柱层析(EA:PE＝1:1)分离纯化得4-(2-溴-5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(19-a,1.44g,2.18mmol,42.3％yield)。MS Calcd:659.15；MS Found:560.21([M-100+H]+).
步骤2：将4-(2-溴-5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(19-a,1.44g,2.18mmol)、3,6-二氢-2H-吡喃-4-硼酸频哪醇酯(687mg,3.27mmol)、Pd(dppf)Cl2·CH2Cl2(357mg,0.44mmol)和磷酸钾(1.39g,6.54mmol)依次加入20mL二氧六环/水(4:1)混合溶液中，抽真空氮气置换3次，升温至80℃搅拌2h。TLC监测反应完成，将反应液完全浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1) 分离纯化得4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(19-b,1.4g,2.11mmol,96.8％yield)。MS Calcd:663.28；MS Found:664.32([M+H]+).
步骤3：将4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(19-b,1.4g,2.11mmol)加入20mL盐酸-二氧六环溶液中，置于室温搅拌过夜。将反应液完全浓缩，向残余物中加入饱和碳酸氢钠溶液，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得粗品2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,0.73g,1.30mmol,61.4％yield)。MS Calcd:563.23；MS Found:564.21([M+H]+).
步骤4：将1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a,80mg,0.41mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,50mg,0.09mmol)加入4mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19,3mg,0.004mmol,6.2％yield)。MS Calcd:713.27；MS Found:714.24([M+H]+).1H NMR(400MHz,DMSO)δ10.06(s,1H),7.79(t,J＝12.8Hz,1H),7.68(t,J＝8.4Hz,1H),7.60(s,1H),6.82(dd,J＝3.2,1.6Hz,1H),5.32(s,2H),4.36(d,J＝12.8Hz,1H),4.29-4.26(m,2H),3.82(t,J＝5.6Hz,2H),3.51-3.17(m,4H),3.05-2.97(m,2H),2.76(d,J＝11.2Hz,2H),2.71–2.69(m,1H),2.38(s,3H),2.36–2.34(m,1H),1.26-1.20(m,5H),1.01–0.94(m,4H).
实施例9 N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物20)的制备
步骤1：将4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2a,550mg，0.83mmol)和2-(4-甲氧基环己-1-烯-1-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼 烷(237mg,1mmol)溶于1,4-二氧六环(8mL)和H2O(4mL)中，再依次加入Pd(dppf)Cl2(60mg,0.09mmol)，磷酸钾(570mg,2.49mmol)，置换三次氮气，80℃反应5.0小时。将反应液浓缩，向残渣中加入30mL乙酸乙酯，经硅藻土过滤，滤液浓缩，经柱层析纯化(DCM:MeOH＝20％:80％)得4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(20-a，370mg，0.53mmol，64％yield)MS Calcd:693.27；MS Found:694.27([M+H]+).
步骤2：将4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(20-a，370mg，0.53mmol)溶于8mL二氯甲烷溶液中，再向其中加入2mL三氟乙酸，室温下搅拌2小时。将反应液直接浓缩，向残渣中加入20mL饱和碳酸氢钠溶液，用二氯甲烷萃取(20mL×3)，合并有机相，干燥浓缩，得到N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(20-b，300mg,0.51mmol,95％yield)MS Calcd:593.21；MS Found:594.27([M+H]+).
步骤3：将N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(20-b，50mg,0.08mmol)和5-羟基-1-甲基-1H-吡唑-4-甲酸(15mg,0.10mmol)置于25mL茄形瓶中，向其中加入5mL DCM溶解，再依次加入PyBOP(43mg,0.08mmol)，DIPEA(0.04mL,0.22mmol)，室温反应3小时。直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(20，6mg,0.01mmol,9％yield)MS Calcd:714.24；MS Found:718.31([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.60(s,1H),10.40(s,1H),8.08(d,J＝8.8Hz,1H),7.98(d,J＝2.4Hz,1H),7.90(s,1H),7.73(dd,J＝8.8,2.4Hz,1H),6.75(t,J＝4.0Hz,1H),5.33(s,2H),4.23(s,2H),3.67(s,3H),3.54-3.43(m,4H),3.30(s,3H),3.12–2.98(m,3H),2.74(d,J＝11.2Hz,2H),2.61–2.41(m,3H),2.20-2.12(m,1H),1.99–1.93(m,1H),1.73-1.63(m,1H),1.21(t,J＝7.2Hz,3H).
实施例10 N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物21)的制备
步骤1：将乙氧基甲叉丙二酸二乙酯(17-a,1.95g,9.04mmol)、异丙基肼盐酸盐(1.0g，9.04mmol)和碳酸钾(2.05g,18.08mmol)分别加入到10mL水溶液中，于100℃条件下搅拌6小时。将反应液冻干，加入少量甲醇溶解，用5N HCl调至pH＝2，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)纯化得5-羟基-1-异丙基-1H-吡唑-4-甲酸乙酯(21-a,1.2g,6.05mmol,67.0％yield)。MS Calcd:198.10；MS Found:199.11([M+H]+).
步骤2：将化合物5-羟基-1-异丙基-1H-吡唑-4-甲酸乙酯(21-a,83mg，0.42mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-6-(哌嗪-1-基)-[1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(20-b,50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟，直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(21，30mg，0.04mmol，47％yield).MS Calcd:745.27；MS Found:746.51([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),8.11-8.08(m,1H),7.99(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.68(s,1H),6.75(t,J＝4.0Hz,1H),5.34(s,2H),4.54-4.47(m,1H),4.36(d,J＝12.8Hz,2H),3.56–3.41(m,3H),3.27(s,3H),3.20-3.13(m,2H),3.02-2.97(m,2H),2.79(d,J＝11.2Hz,2H),2.62-2.47(m,4H),2.21-2.13(m,1H),2.00-1.94(m,1H),1.73–1.64(m,1H),1.37(d,J＝6.8Hz,6H),1.23(t,J＝7.2Hz,3H).
实施例11 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物22)的制备
步骤1：将(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,60mg,0.10mmol)，DIPEA(13mg，0.10mmol)，5-羟基-1-异丙基-1H-吡唑-4-甲酸乙酯(21-a,102mg，0.5mmol)加入到2-甲基-2-丁醇(1mL)中，油浴升温到180℃，搅拌30分钟，将反应体系溶解到甲醇中，直接prep-HPLC纯化，得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(22,17mg,0.023mmol,23.2％yield)。MS Calcd:731.26；MS Found:732.31([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.09(d,J＝8.8Hz,1H),7.99(d,J＝2.0Hz,1H),7.75(dd,J＝8.8,2.0Hz,1H),7.67(s,1H),6.89-6.85(m,1H),5.35(d,J＝2.4Hz,2H),4.71-4.63(m,1H),4.54-4.47(m,1H),4.32-4.26(m,3H),3.83(t,J＝5.6Hz,2H),3.71(d,J＝11.2Hz,1H),3.52(t,J＝11.2Hz,1H),3.22-3.15(m,1H),2.99-2.92(m,1H),2.80(d,J＝10.0Hz,1H),2.68(d,J＝10.0Hz,1H),2.55-2.51(m,3H),1.45(brs,3H),1.36(d,J＝6.8Hz,6H),1.25(t,J＝7.2Hz,3H).
实施例12 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-2-甲基喹啉-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物26)的制备
步骤1：称取3-羟基-2-甲基喹啉-4-甲酸(26-a,19mg,0.09mmol)，PyBOP(46.8mg,0.09mmol)，N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,60mg,0.09mmol)到25mL单口瓶中，加入DCM(2mL)和DIPEA(0.04mL,0.27mmol)，加完后室温搅拌过夜。将反应液减压浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-2-甲基喹啉-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(26,19mg,0.02mmol,26.9％Yeild)。MS Calcd:750.23；MS Found:751.12([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),8.08(d,J＝8.8Hz,1H),7.97(d,J＝2.0Hz,1H),7.91–7.87(m,1H),7.73(dd,J＝8.8,2.0Hz,1H),7.68–7.65(m,1H),7.59–7.47(m,2H),6.85–6.83(m,1H),5.33(s,2H),4.72(d,J＝12.8Hz,1H),4.27(t,J＝2.8Hz,2H),3.82(t,J＝5.2Hz,2H),3.60–3.54(m,1H),3.34–2.83(m,7H),2.63-2.53(m,6H),1.20(t,J＝7.2Hz,3H).
实施例13 2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物27)的制备
步骤1：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,80mg,0.38mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,50mg,0.09mmol)加入4mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(27,10mg,0.01mmol,14.6％yield)。MS Calcd:727.29；MS Found:728.32([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.09(s,1H),7.81(d,J＝12.8Hz,1H),7.70-7.68(m,3H),6.85–6.83(m,1H),5.32(s,2H),4.84-4.74(m,1H),4.36-4.26(m,4H),3.82(t,J＝5.2Hz,2H),3.52–3.17(m,6H),3.05-2.99(m,2H),2.78(d,J＝11.2Hz,2H),2.38(s,3H),2.35–2.27(m,2H),1.84-1.75(m,2H),1.26–1.20(m,5H).
实施例14 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物28)的制备
步骤1：将5-羟基-1-甲基-1H-吡唑-4-甲酸(3-c,40mg,0.28mmol)和1-氯-N,N,2-三甲基丙烯胺(42mg,0.31mmol)分别加入到5mL二氯甲烷溶液中，室温下搅拌3小时，然后加入2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,80mg,0.14mmol)、DIPEA(90mg,0.71mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩后残余物溶于4mL甲醇/水(5:1)混合溶剂中，然后加入氢氧化钠(15mg,0.37mmol)，置于室温搅拌过夜。将反应液调至pH＝2，经prep-HPLC得标题化合物：2-(2-(3,6- 二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-4-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(28，10mg,0.01mmol,11.2％yield)。MS Calcd:687.25；MS Found:688.42([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.27(s,1H),8.43(s,1H),7.90(s,1H),7.81(d,J＝12.8Hz,1H),7.68(d,J＝8.0Hz,1H),6.84(s,1H),5.34(s,2H),4.28-4.26(m,2H),3.82(t,J＝5.6Hz,2H),3.68(s,3H),3.56–2.98(m,10H),2.75(d,J＝11.2Hz,2H),2.38-2.34(m,3H),1.23(t,J＝7.2Hz,3H).
实施例15 2-(6-(4-(5-氯-3-羟基-1-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(化合物29)的制备
步骤1：将3-羟基-1-甲基-1H-吡唑-4-甲酸乙酯(16-a,500mg,2.94mmol)溶于10mL二氯甲烷溶液中，置于0℃条件下缓慢加入磺酰氯(0.47g，3.53mmol)，缓慢升至室温搅拌过夜。向反应液中加入水中终止反应，二氯甲烷萃取，减压浓缩，残余物经prep-HPLC纯化得5-氯-3-羟基-1-甲基-1H-吡唑-4-甲酸乙酯(29-a,120mg,0.59mmol,20％yield)。MS Calcd:204.03；MS Found:205.10([M+H]+).
步骤2：将5-氯-3-羟基-1-甲基-1H-吡唑-4-甲酸乙酯(29-a,120mg,0.59mmol)溶于4mL甲醇/水(5：1)混合溶液中，加入氢氧化钾(200mg,3.52mmol)，升温至回流搅拌过夜，将反应液完全浓缩，向残余物中缓慢加入5N HCl溶液，直至固体析出，抽滤得5-氯-3-羟基-1-甲基-1H-吡唑-4-甲酸(29-b,90mg,0.51mmol,86.9％yield)。MS Calcd:176.00；MS Found:177.01([M+H]+).
步骤3：将5-氯-3-羟基-1-甲基-1H-吡唑-4-甲酸(29-b,44mg,0.25mmol)和1-氯-N,N,2-三甲基丙烯胺(36mg,0.27mmol)分别加入到5mL二氯甲烷溶液中，室温下搅拌3小时，然后加入N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,70mg,0.12mmol)、DIPEA(80mg,0.62mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：2-(6-(4-(5-氯-3-羟基-1-甲基-1H-吡唑-4- 羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(29，16mg,0.02mmol,18％yield)。MS Calcd:723.17；MS Found:724.30([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.75(s,1H),10.38(s,1H),8.08(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),6.86–6.84(m,1H),5.33(s,2H),4.27(q,J＝2.8Hz,2H),3.82(t,J＝5.2Hz,2H),3.64(s,3H),3.49-3.42(m,2H),3.04-2.98(m,2H),2.75–2.68(m,2H),2.54–2.51(m,6H),1.21(t,J＝7.2Hz,3H).
实施例16 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物30)的制备
步骤1将2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,50mg,0.09mmol)和5-羟基-1-异丙基-1H-吡唑-4-甲酸乙酯(21-a,87.9mg,0.44mmol)加入4mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(30,10mg,0.01mmol,15％yield)。MS Calcd:715.29；MS Found:716.30([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.09(s,1H),7.81(d,J＝12.8Hz,1H),7.70–7.67(m,2H),6.85–6.82(m,1H),5.32(s,2H),4.54-4.47(m,1H),4.36(d,J＝12.8Hz,2H),4.27(q,J＝2.8Hz,2H),3.82(t,J＝5.6Hz,2H),3.52–3.19(m,6H),3.02(q,J＝7.2Hz,2H),2.79(d,J＝11.2Hz,2H),2.37(s,3H),1.36(d,J＝6.8Hz,6H),1.23(t,J＝7.2Hz,3H).
实施例17 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物31)的制备
步骤1：将5-羟基-1-异丙基-1H-吡唑-4-甲酸乙酯(21-a,90mg,0.44mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,50mg,0.09mmol)加入4mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(31,10mg,0.01mmol,15.7％yield)。MS Calcd:717.24；MS Found:718.21([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.08(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.63(s,1H),6.86–6.84(m,1H),5.35(s,2H),4.53-4.46(m,1H),4.37(d,J＝12.8Hz,2H),4.27(d,J＝3.2Hz,2H),3.82(t,J＝5.6Hz,2H),3.52–3.18(m,6H),3.03(q,J＝7.2Hz,2H),2.78(d,J＝11.2Hz,2H),1.34(d,J＝6.8Hz,6H),1.23(t,J＝7.2Hz,3H).
实施例18 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物32)的制备
步骤1：将(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,55mg,0.10mmol)，DIPEA(13mg，0.10mmol)，1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,63mg，0.3mmol)加入到2-甲基-2-丁醇中，油浴升温到180℃，搅拌30分钟，将反应体系溶解到甲醇中，直接prep-HPLC纯化，得到目标化合物(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(32,7mg,0.0094mmol,9.4％yield)。MS Calcd:743.26；MS Found:744.30([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.08(d,J＝8.8Hz,1H),7.99(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.70(s,1H),6.85(dd,J＝3.6,2.0Hz,1H),5.35(s,2H),4.84-4.76(m,1H),4.29-4.26(m,2H),4.19-4.13(m,1H),4.10-4.04(m,1H),3.82(t,J＝5.2Hz,2H),3.71(d,J＝11.2Hz,1H),3.53–3.14(m,4H),2.96-2.93(m,1H),2.80(d,J＝10.2Hz,1H),2.34–2.29(m,2H),1.94-1.88(m,1H),1.84–1.78(m,3H),1.66–1.57(m,1H),1.46-1.42(m,2H),1.26–1.19(m,5H).
实施例19 (R)-2-(6-(4-(5-氯-3-羟基-1-甲基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(化合物33)的制备
步骤1：将5-氯-3-羟基-1-甲基-1H-吡唑-4-甲酸(29-b,30mg,0.17mmol)加入到DCM中(1mL)，然后加入1-氯-N,N,2-三甲基丙基-1-烯-1-胺(23mg，0.17mmol)，室温下搅拌2小时，然后加入到(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,40mg,0.07mmol)和DIPEA(54mg，0.42mmol)的二氯甲烷溶液中，并在室温下搅拌过夜，然后加入水(3mL)，用DCM萃取(3mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，剩余物经Prep-HPLC纯化，得到标题化合物：(R)-2-(6-(4-(5-氯-3-羟基-1-甲基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(33,7mg,0.009mmol,13.5％yield)。MS Calcd:737.19；MS Found:738.21([M+H]+)。1H NMR(400MHz,DMSO-d6)δ8.01(d,J＝8.8Hz,1H),7.94(d,J＝2.0Hz,1H),7.71(dd,J＝8.8,2.0Hz,1H),6.83(t,J＝2.0Hz,1H),5.29(s,2H),4.25(t,J＝2.8Hz,2H),3.67–3.45(m,10H),3.13-2.87(m,3H),2.76-2.63(m,3H),1.26–1.8(m,6H).
实施例20 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物34)的制备
步骤1：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,88mg,0.42mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(20-b，50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直 接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(34，25mg，0.03mmol，39％yield).MS Calcd:757.27；MS Found:758.41([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),7.99(d,J＝8.0Hz,1H),7.93(d,J＝2.0Hz,1H),7.69(dd,J＝8.8,2.0Hz,1H),7.61(s,1H),6.72(t,J＝4.0Hz,1H),5.27(s,2H),4.79-4.73(m,1H),4.28–4.25(m,2H),4.13-4.01(m,1H),3.52–3.38(m,3H),3.25(s,3H),3.13-2.96(m,4H),2.74(d,J＝10.8Hz,1H),2.47-2.39(m,3H),2.24 2.11(m,2H),1.93–1.59(m,6H),1.20–1.12(m,5H).
实施例21 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-2-甲基喹啉-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物35)的制备
步骤1：将4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-1,200mg,0.47mmol)置于100mL茄形瓶中，加入1,4-二氧六环溶液10mL。再向其中依次加入N-(3-氟-2-甲基-4-(三氟甲基)苯基)-2-碘代乙酰胺(int-4,185.9mg,0.51mmol)和DIPEA(0.23mL,1.4mmol)。75℃下搅拌5小时。直接将反应液浓缩，经柱层析纯化(PE:EA＝60％:40％)得4-(2-溴-5-乙基-4-(2-(3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(35-a，180mg,0.27mmol,58％yield).MS Calcd:661.15；MS Found:662.30([M+H]+).
步骤2：将4-(2-溴-5-乙基-4-(2-(3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(35-a，180mg,0.27mmol)和2-(3,6-二氢-2H-吡喃-4-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(85.9mg,0.41mmol)溶于1,4-二氧六环(6mL)和H2O(3mL)中，再依次加入Pd(dppf)Cl2(20mg,0.03mmol)，磷酸钾(188mg,0.82mmol)，置换三次氮气，80℃反应5.0小时。将反应液浓缩，向残渣中加入30mL乙酸乙酯，经硅藻土过滤， 滤液浓缩，经柱层析纯化(DCM:MeOH＝4:1)得4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(2-((3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(35-b，153mg，0.23mmol，85％yield)MS Calcd:663.28；MS Found:664.20([M+H]+).
步骤3：将4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(2-((3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(35-b，153mg，0.23mmol)溶于8mL二氯甲烷溶液中，再向其中加入2mL三氟乙酸，室温下搅拌2小时。将反应液直接浓缩，向残渣中加入20mL饱和碳酸氢钠溶液，用二氯甲烷萃取(20mL×3)，合并有机相，干燥浓缩，得到2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(35-c，150mg，0.27mmol，98％yield)MS Calcd:563.23；MS Found:564.31([M+H]+).
步骤4：将化合物3-羟基-2-甲基喹啉-4-甲酸(27.0mg,0.13mmol)和HATU(102.8mg,0.27mmol)置于25mL茄形瓶中，向其中加入5mLDCM，然后加入DIPEA(0.04mL,0.27mmol)，加完后搅拌2分钟后。再加入2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(35-c，50mg,0.09mmol)，室温反应3小时。直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-2-甲基喹啉-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(35，13mg，0.02mmol，19.6％yield)MS Calcd:748.27；MS Found:749.41([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.25(s,1H),9.89(s,1H),7.91–7.88(m,1H),7.67–7.45(m,5H),6.82(s,1H),5.22(s,2H),4.70–4.66(m,1H),4.25(s,2H),3.83(t,J＝5.2Hz,2H),3.59–3.53(m,1H),3.31-3.04(m,4H),2.99-2.84(m,4H),2.63-2.60(m,5H),2.25(s,3H),1.17(t,J＝7.2Hz,3H).
实施例22 2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物36)的制备
步骤1：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,88mg,0.42mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶 -4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(35-c，50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，经Prep-HPLC纯化得标题化合物：2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(36，24mg，0.03mmol，37％yield).MS Calcd:727.29；MS Found:728.40([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.23(s,1H),7.67(s,1H),7.59-7.50(m,2H),6.82(s,1H),5.24(s,2H),4.79-4.68(m,1H),4.27–4.23(m,4H),3.79(t,J＝5.6Hz,2H),3.45–3.38(m,2H),3.18–3.12(m,2H),3.01-2.95(m,2H),2.75(d,J＝10.8Hz,2H),2.50-2.41(m,4H),2.30–2.25(m,2H),2.20(d,J＝2.0Hz,3H),1.79-1.71(m,2H),1.18(t,J＝7.2Hz,3H).
实施例23 N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(5-羟基-1-(2,2,2-三氟乙基)-1H-吡唑-4-酰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物37)的制备
步骤1：将(2,2,2-三氟乙基)肼盐酸盐(37-a，4000mg，35.06mmol)溶于40mL水中，然后在0℃下缓慢加入2-(乙氧基亚甲基)丙二酸二乙酯(7579.3mg,35.06mmol)和K2CO3(9675mg,17.53mmol)，然后升至100℃搅拌5h。TLC检测原料反应完毕。反应液冷却至室温，旋干部分溶剂，残余物用5N HCl调至pH＝2，二氯甲烷萃取(50mL×3)，饱和食盐水洗涤(100mL)，合并有机相，干燥，滤液减压浓缩，残余物经柱层析纯化(DCM:MeOH＝20％:80％)得到5-羟基-1-(2,2,2-三氟乙基)-1H-吡唑-4-甲酸乙酯(37-b，3500mg，42％yield)MS Calcd:238.06；MS Found:239.10([M+H]+).
步骤2：将5-羟基-1-(2,2,2-三氟乙基)-1H-吡唑-4-甲酸乙酯(37-b，100mg,0.44mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(20-b，50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，经Prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(5-羟基-1-(2,2,2-三氟乙基)-1H-吡唑-4-酰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(37，20mg,0.02mmol,97％yield，消旋体).MS Calcd:785.23；MS Found:786.37([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.04(d,J＝8.8Hz,1H),7.95(d,J＝2.0 Hz,1H),7.73-7.70(m,2H),6.73(d,J＝4.0Hz,1H),5.30(s,2H),4.79(q,J＝8.8Hz,2H),4.25(d,J＝12.0Hz,2H),3.54–3.41(m,3H),3.27(s,3H),3.21-3.12(m,2H),2.99(q,J＝7.2Hz,2H),2.76(d,J＝10.8Hz,2H),2.58-2.44(m,3H),2.18–2.11(m,1H),1.95–1.91(m,1H),1.72–1.62(m,1H),1.20(t,J＝7.2Hz,3H).
中间体38-f的制备：N-(2-氯-5-甲基-3-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺
步骤1：称取5-氯-2-甲基-4-(三氟甲基)苯胺(38-a,6.0g,28.63mmol)到50mL单口瓶中，加入DCM(30mL)，然后加入三乙胺(7.94mL,57.25mmol)，冰浴下滴加氯乙酰氯(3.40mL,34.35mmol)的DCM溶液(10mL)，然后自然恢复室温反应过夜。浓缩反应液，剩余物快速柱层析(PE:EA＝20:1)得2-氯-N-(5-氯-2-甲基-4-(三氟甲基)苯基)乙酰胺(38-b,3.7g,12.93mmol,45.1％yield)。MS Calcd:284.99；MS Found:283.97([M-H]-).
步骤2：向100mL茄形瓶中加入2-氯-N-(5-氯-2-甲基-4-(三氟甲基)苯基)乙酰胺(38-b,3.7g,12.93mmol)和溶剂丙酮(15mL)。然后向其中加碘化钾(5.4g,32.33mmol)，50℃搅拌3小时。将溶剂减压旋干得粗品，粗品通过柱层析纯化(PE:EA＝3:1)得N-[5-氯-2-甲基-4-(三氟甲基)苯基]-2-碘代乙酰胺(38-c,3.4g,9.01mmol,69.4％yield)。MS Calcd:376.93；MS Found:375.97([M-H]-).
步骤3：向100mL茄形瓶中加入N-[5-氯-2-甲基-4-(三氟甲基)苯基]-2-碘代乙酰胺(38-c,1.1g,2.81mmol)，4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-1,1.0g,2.34mmol)和溶剂1,4-二氧六环(15mL)。然后向其中加DIPEA(0.9g,7.02mmol)，90℃搅拌2小时。将溶剂减压旋干得粗品，粗品通过柱层析纯化(PE:EA＝1:4)得到4-(2-溴-4-(2-(2-氯-5-甲基-3-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(38-d,940mg,1.39mmol,59.3％yield)。MS Calcd:676.92.；MS Found:578.12([M+H-100]+).
步骤4：向100mL茄形瓶中加入4-(2-溴-4-(2-(2-氯-5-甲基-3-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1- 甲酸叔丁酯(38-d,940mg,1.39mmol)和溶剂1,4-二氧六环(10mL)，H2O(5mL)。然后向其中加2-(3,6-二氢-2H-吡喃-4-基)-4,4,5,5-四甲基-1,3,2-二氧杂硼烷(438mg,2.08mmol)，Pd(dppf)Cl2(102mg,0.14mmol),磷酸钾(960mg,4.17mmol)，氮气保护下100℃搅拌6小时。LCMS检测原料反应完毕，向其中加入水(50mL)，用乙酸乙酯萃取(50mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得粗品，粗品通过柱层析纯化(DCM:MeOH＝30:1)得4-(4-(2-(2-氯-5-甲基-3-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(38-e,800mg,1.18mmol,84.7％yield)MS Calcd:679.25.；MS Found:624.21([M+H-56]+).
步骤5：向100mL茄形瓶中加入4-(4-(2-(2-氯-5-甲基-3-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(38-e,800mg,1.18mmol)和溶剂DCM(5mL)。然后向其中加TFA(0.5mL)，室温搅拌过夜。向其中加入H2O(20mL)，用饱和碳酸氢钠溶液将pH调至碱性，用二氯甲烷萃取(30mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得到N-(2-氯-5-甲基-3-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(38-f,500mg,0.86mmol,73.3％yield)MS Calcd:579.20.；MS Found:580.22([M+H]+).
实施例24 N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物39)的制备
步骤1：将5-羟基-1-异丙基-1H-吡唑-4-甲酸乙酯(21-a,85mg,0.43mmol)和N-(2-氯-5-甲基-3-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(38-f,50mg,0.09mmol)加入4mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(39,8mg,0.01mmol,12.3％yield)。MS Calcd:731.26；MS Found:732.26([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.12(s,1H),7.98(s,1H),7.78(s,1H),7.67(s,1H),6.84(t,J＝2.4Hz,1H),5.30(s,2H),4.53-4.46(m,1H),4.37(d,J＝12.4Hz, 2H),4.28(d,J＝2.8Hz,2H),3.82(t,J＝5.2Hz,2H),3.52–3.17(m,6H),3.06-3.00(m,2H),2.79(d,J＝11.2Hz,2H),2.38(s,3H),1.35(d,J＝6.8Hz,6H),1.20(t,J＝7.2Hz,3H).
实施例25 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物40)的制备
步骤1：将5-羟基-1-甲基-1H-吡唑-3-甲酸甲酯(40-a,120mg,0.77mmol)溶于3mL甲醇/水(5:1)混合溶液中，加入氢氧化钾(258mg,4.61mmol)，升温至回流搅拌过夜，将反应液完全浓缩，向残余物中缓慢加入5N HCl溶液，调至pH＝2，减压浓缩得5-羟基-1-甲基-1H-吡唑-3-甲酸(40-b,100mg,0.7mmol,91.6％yield)。MS Calcd:142.04；MS Found:143.06([M+H]+).
步骤2：将5-羟基-1-甲基-1H-吡唑-3-甲酸(40-b,34mg,0.24mmol)和1-氯-N,N,2-三甲基丙烯胺(36mg,0.27mmol)分别加入到5mL二氯甲烷溶液中，室温下搅拌3小时，然后加入N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,70mg,0.12mmol)、DIPEA(80mg,0.62mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，减压浓缩，经Prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-甲基-1H-吡唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(40，6mg,0.01mmol,7％yield)。MS Calcd:689.21；MS Found:690.18([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.36(s,1H),8.00(d,J＝8.8Hz,1H),7.93(d,J＝2.0Hz,1H),7.70(dd,J＝8.8,2.0Hz,1H),6.84–6.82(m,1H),5.29(s,2H),4.78(s,1H),4.46(s,1H),4.25(t,J＝2.8Hz,2H),3.80(t,J＝5.6Hz,2H),3.51(s,3H),3.44–3.38(m,2H),3.29-3.23(m,1H),3.00-2.91(m,4H),2.74–2.68(m,3H),1.19(t,J＝7.2Hz,3H).
实施例26 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物41)的制备
步骤1：称取1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,65mg,0.30mmol)，N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,40mg,0.06mmol)到25mL单口瓶中，加入2-甲基-1-丁醇(2mL)和DIPEA(0.01mL,0.06mmol)，加完后升温到180摄氏度搅拌30分钟。反应液减压浓缩后经prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(41,10mg,0.01mmol,21.0％Yeild)。MS Calcd:743.26；MS Found:744.36([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.08(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.68(s,1H),6.85(t,J＝2.0Hz,1H),5.34(s,2H),4.70–4.59(m,1H),4.35(d,J＝12.6Hz,2H),4.27(d,J＝3.0Hz,2H),3.82(t,J＝5.2Hz,2H),3.50-3.18(m,3H),3.02(q,J＝7.2Hz,2H),2.79(d,J＝11.1Hz,2H),2.02–1.77(m,8H),1.66–1.58(m,3H),1.23(t,J＝7.2Hz,3H).
实施例27 2-(6-(4-(1-(叔丁基)-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物42)的制备
步骤1：将乙氧基甲叉丙二酸二乙酯(17-a,1.74g,8.03mmol)、特丁基肼盐酸盐(1.0g，8.03mmol)和碳酸钾(2.21g,16.05mmol)分别加入到10mL水溶液中，于100℃条件下搅拌6小时。用5N HCl调至pH＝2，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得1-(叔丁基)-5-羟基-1H-吡唑-4-甲酸乙酯(42-a,0.45g,2.12mmol,26.4％yield)。MS Calcd:212.12；MS Found:213.21([M+H]+).
步骤2：将1-(叔丁基)-5-羟基-1H-吡唑-4-甲酸乙酯(42-a,94mg,0.44mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a] 嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,50mg,0.09mmol)加入2mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：2-(6-(4-(1-(叔丁基)-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(42,6mg,0.01mmol,9.1％yield)。MS Calcd:729.30；MS Found:730.27([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.09(s,1H),7.81(d,J＝12.8Hz,1H),7.73–7.68(m,2H),6.85–6.83(m,1H),5.32(s,2H),4.41(d,J＝12.8Hz,2H),4.28(d,J＝2.8Hz,2H),3.82(t,J＝5.6Hz,2H),3.53-3.46(m,2H),3.27-3.22(m,2H),3.03(q,J＝7.2Hz,2H),2.81(d,J＝11.2Hz,2H),2.55-2.51(m,2H),2.38(s,3H),1.57(s,9H),1.23(t,J＝7.2Hz,3H).
实施例28 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-(1,1,1-三氟丙烷-2-基)-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物43)的制备
步骤1：将1,1,1-三氟丙酮(43-a,4.9g,44.05mmol)和苯甲酰肼(43-b,4.0g,29.37mmol)依次加入40mL甲苯溶液中，置于闷罐中，升温至110℃搅拌过夜。将反应液完全浓缩，残余物经硅胶柱层析(PE:EA＝10:1)分离纯化得N'-(1,1,1-三氟丙烷-2-亚基)苯甲酰肼(43-c,5.1g,22.15mmol,75.4％yield)。MS Calcd:230.07；MS Found:231.20([M+H]+).
步骤2：将N'-(1,1,1-三氟丙烷-2-亚基)苯甲酰肼(43-c,3.0g,13.3mmol)溶于40mL四氢呋喃溶液中，置于0℃条件下缓慢滴加硼烷四氢呋喃溶液(26.06mL,26.06mmol)，滴毕，缓慢升至室温搅拌过夜。将反应液再次冷却至0℃，加入5mL甲醇，将反应液完全浓缩，向残余物中加入水，二氯甲烷萃取，饱和氯化铵溶液洗涤，减压浓缩得粗品N'-(1,1,1-三氟丙-2-基)苯并酰肼(43-d,2.8g,12.06mmol,92.5％yield)。产物未经纯化直接投入下一步反应。MS Calcd:232.08；MS Found:233.1([M+H]+).
步骤3：将N'-(1,1,1-三氟丙-2-基)苯并酰肼(43-d,1.0g,4.31mmol)溶于5mL甲醇溶液中，然后加入8mL浓盐酸，升温至100℃搅拌过夜。TLC监测原料 消失，将反应液完全浓缩得粗产物(1,1,1-三氟丙-2-基)肼盐酸盐(43-e,1.5g,9.2mmol)。
步骤4：将乙氧基甲叉丙二酸二乙酯(17-a,1.0g,94.68mmol)、(1,1,1-三氟丙-2-基)肼盐酸盐(43-e,1.5g,9.2mmol)和碳酸钾(2.6g,18.74mmol)分别加入到10mL水溶液中，于100℃条件下搅拌6小时。用5N HCl调至pH＝2，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得5-羟基-1-(1,1,1-三氟丙-2-基)-1H-吡唑-4-甲酸乙酯(43-f,0.56g,2.2mmol,23.7％yield)。MS Calcd:252.07；MS Found:253.16([M+H]+).
步骤5：将5-羟基-1-(1,1,1-三氟丙-2-基)-1H-吡唑-4-甲酸乙酯(43-f,140mg,0.54mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,50mg,0.09mmol)加入4mL 2-甲基-2-丁醇溶液中，置于微波反应器中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-(1,1,1-三氟丙烷-2-基)-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(43,5mg,0.01mmol,6.5％yield)。MS Calcd:769.26；MS Found:770.28([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.09(s,1H),7.81(d,J＝12.8Hz,1H),7.71–7.68(m,2H),6.85–6.83(m,1H),5.32(s,2H),4.35–4.27(m,4H),3.82(t,J＝5.6Hz,2H),3.53–3.17(m,4H),3.02(q,J＝7.2Hz,2H),2.78(d,J＝11.2Hz,2H),2.55-2.51(m,2H),2.38(s,3H),1.61(d,J＝7.2Hz,3H),1.23(t,J＝7.2Hz,3H).
实施例29 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3R)-4-(5-羟基-1-(1,1,1-三氟丙基-2-基)-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物44)的制备
步骤1：将(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,50mg,0.09mmol)，DIPEA(13mg，0.10mmol)，5-羟基-1-(1,1,1-三氟丙-2-基)-1H-吡唑-4-甲酸乙酯(43-f,126mg，0.5mmol)加入到2-甲基-2-丁醇(1mL)中，油浴升温到180℃，搅拌30分钟，将反应体系溶解到甲醇中，直接prep-HPLC纯化，得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙 基-6-(3R)-4-(5-羟基-1-(1,1,1-三氟丙基-2-基)-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(44,12mg,0.0152mmol,17.0％yield)。MS Calcd:785.23；MS Found:786.31([M+H]+)。1H NMR(400MHz,DMSO-d6+重水)δ8.02(d,J＝8.8Hz,1H),7.95(d,J＝2.0Hz,1H),7.75(s,1H),7.72(dd,J＝8.8,2.0Hz,1H),6.84–6.82(m,1H),5.30(d,J＝2.4Hz,2H),4.25(d,J＝3.2Hz,2H),3.66(d,J＝11.2Hz,2H),3.81-3.76(m,2H),3.48(t,J＝11.2Hz,2H),3.16-3.11(m,2H),2.95-2.85(m,2H),2.79(d,J＝9.6Hz,2H),2.55-2.51(m,2H),1.61(d,J＝7.2Hz,3H),1.43(d,J＝9.2Hz,3H),1.22(t,J＝7.2Hz,3H).
实施例30 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-3-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物45)的制备
步骤1：将2-(乙氧基羰基)-3-氧代丁酸乙酯(45-a,1g,4.95mmol)和异丙基肼盐酸盐(45-b,0.4g,4.95mmol)溶于EtOH(20mL)中，加入HCl(1mL,4.95mmol),85℃反应2小时。将溶剂旋干后加入30mL水，用二氯甲烷萃取(30mL*3)，有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得到粗品经柱层析纯化(DCM:MeOH＝30:1)得5-羟基-1-异丙基-3-甲基-1H-吡唑-4-甲酸乙酯(45-c,500mg,2.36mmol,47.6％yield)。MS Calcd:212.12；MS Found:213.22([M+H]+).
步骤2：将5-羟基-1-异丙基-3-甲基-1H-吡唑-4-甲酸乙酯(45-c,104mg,0.45mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,50mg,0.09mmol)加入4mL 2-甲基-2-丁醇溶液中，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-3-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(45,8mg,0.01mmol,11.9％yield)。MS Calcd:729.30；MS Found:730.47([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.09(s,1H),7.81(d,J＝12.8Hz,1H),7.68(d,J＝8.0Hz,1H),6.85–6.82(m,1H),5.31(s,2H),4.46-4.41(m,1H),4.28-4.26(m,2H),4.12(brs,2H),3.82(t,J＝5.6Hz,2H),3.51-3.44(m,2H),3.09-2.98(m,5H),2.73–2.69(m,3H),2.37(s,3H),2.16(s,3H),1.29–1.19(m,9H).
实施例31 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6- (4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物47)的制备
步骤1：将4-氯乙酰乙酸乙酯(47-a,2.2g,13.37mmol)溶于10mL乙酸中，置于0℃条件下，缓慢加入亚硝酸钠(1.2g,16.71mmol)的水溶液(10mL)，加入完毕缓慢升至室温搅拌过夜。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析分离纯化得4-氯-2-(羟基亚氨基)-3-氧代丁酸乙酯(47-b,1300mg,6.71mmol,50.2％yield).1H NMR(400MHz,DMSO)δ13.60(s,1H),4.91(s,2H),4.28(q,J＝7.2Hz,2H),1.24(t,J＝7.2Hz,3H).
步骤2：将4-氯-2-(羟基亚氨基)-3-氧代丁酸乙酯(47-b,500mg,2.58mmol)加入3mL DMF溶液中，然后加入尿素(1.24g,20.66mmol)，于100℃条件下搅拌3h。向反应液中加入水，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(PE:EA＝2:1)分离纯化得4-羟基异噁唑-3-甲酸乙酯(47-c,210mg,1.34mmol,51.8％yield)。1H NMR(400MHz,DMSO)δ9.72(s,1H),8.71(s,1H),4.35(q,J＝7.2Hz,2H),1.31(t,J＝7.2Hz,3H).
步骤3：将4-羟基异噁唑-3-甲酸乙酯(47-c,70mg,0.44mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,50mg,0.09mmol)加入4mL 2-甲基-2-丁醇溶液中，置于油浴中升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(47,4mg,0.01mmol,6.7％yield)。MS Calcd:676.18；MS Found:677.2([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.34(s,1H),8.59(s,1H),8.07(d,J＝8.8Hz,1H),7.97(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),6.85(t,J＝2.0Hz,1H),5.34(s,2H),4.53(d,J＝12.4Hz,1H),4.26(t,J＝2.8Hz,2H),3.82(t,J＝5.2Hz,2H),3.63(d,J＝12.4Hz,1H),3.51-3.36(m,4H),3.06-2.97(m,4H),2.84(d,J＝11.2Hz,1H),2.73(d,J＝11.2Hz,1H),1.21(t,J＝7.2Hz,3H).
实施例32 N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物48)的制备
步骤1：向50mL茄形瓶中加入N-(2-氯-5-甲基-3-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(38-f,50mg,0.09mmol)和溶剂2-甲基-2-丁醇(2mL)。然后向其中加1-环丁基-5-羟基吡唑-4-甲酸乙酯(17-b,91mg,0.45mmol)，DIPEA(0.04mL,0.27mmol)，180℃搅拌30分钟。向其中加入甲醇(1.5mL)，粗品通过Pre-HPLC纯化得到标题化合物：N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(48,10mg,0.01mmol,14.5％yield)。MS Calcd:743.26；MS Found:744.31([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.13(s,1H),7.98(s,1H),7.78(s,1H),7.67(s,1H),6.85–6.84(m,1H),5.30(s,2H),4.82–4.75(m,1H),4.35(d,J＝12.8Hz,2H),4.28(d,J＝2.8Hz,2H),3.82(t,J＝5.6Hz,2H),3.57–3.18(m,4H),3.03(q,J＝7.2Hz,2H),2.78(d,J＝11.2Hz,2H),2.54-2.45(m,2H),2.38(s,3H),2.32–2.19(m,2H),1.83-1.71(m,2H),1.22-1.17(m,5H).
实施例33 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-3-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰基(化合物49)的制备
步骤1：向50mL茄形瓶中加入化合物N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,100mg,0.18mmol)和溶剂2-甲基-2-丁醇(1.5mL)。然后向其中加5-羟基-3-甲基-1-(丙-2-基)吡唑-4-甲酸乙酯(45-c,49mg,0.23mmol)，DIPEA(0.09mL,0.53mmol)，180℃搅拌30分钟。减压浓缩后粗品通过Pre-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-异丙基-3-甲基-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰基(49,13mg,0.02mmol,9.4％yield)MS Calcd:731.26；MS Found:732.30([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.39(s,1H),8.08(d, J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),6.86–6.84(m,1H),5.34(s,2H),4.46-4.39(m,1H),4.27(q,J＝2.8Hz,2H),4.13(d,J＝12.4Hz,2H),3.82(t,J＝5.6Hz,2H),3.51–3.44(m,2H),3.09–2.99(m,4H),2.72–2.68(m,2H),2.54–2.51(m,2H),2.15(s,3H),1.28–1.19(m,9H).
实施例34 N-[2-氯-6-(三氟甲基)吡啶-3-基]-2-(6-{4-[(1-环丁基-5-羟基吡唑-4-基)羰基]哌嗪-1-基}-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)乙酰胺(化合物51)的制备
步骤1：向250mL茄形瓶中加入化合物2-氯-6-(三氟甲基)吡啶-3-胺(51-a,4.8g,24.42mmol)和溶剂二氯甲烷(30mL)。然后向其中加三乙胺(4.9g,48.83mmol)和氯乙酰氯(2.8g,24.42mmol)，室温搅拌过夜。加入150mL二氯甲烷稀释，用水洗涤有机相(100mL*2)，再用无水硫酸钠干燥，将有机相减压旋干得粗品，粗品通过柱层析纯化(PE:EA＝4:1)得到2-氯-N-[2-氯-6-(三氟甲基)吡啶-3-基]乙酰胺(51-b,1.8g,6.41mmol,26.3％yield).MS Calcd:271.97.；MS Found:270.97([M-H]-).
步骤2：向100mL茄形瓶中加入2-氯-N-[2-氯-6-(三氟甲基)吡啶-3-基]乙酰胺(51-b,1.8g,6.41mmol)和溶剂丙酮(30mL)。然后向其中加碘化钾(2.6g,16.03mmol)，50℃搅拌3小时。将溶剂减压旋干得粗品，粗品通过柱层析纯化(PE:EA＝4:1)得到N-[2-氯-6-(三氟甲基)吡啶-3-基]-2-碘代乙酰胺(51-c,1.8g,4.86mmol,75.8％yield)。MS Calcd:363.91.；MS Found:362.91([M-H]-).
步骤3：向100mL茄形瓶中加入化合物N-[2-氯-6-(三氟甲基)吡啶-3-基]-2-碘代乙酰胺(51-c,457mg,1.25mmol)，4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-5,450mg,1.05mmol)和溶剂1,4-二氧六环(15mL)。然后向其中加DIPEA(405mg,3.15mmol)，60℃搅拌过夜。将溶剂减压旋干得粗品，粗品通过柱层析纯化(PE:EA＝1:3)得到4-(4-(2-(2-氯-6-(三氟甲基)吡啶-3-基)氨基)-2-氧乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(51-d,650mg,0.97mmol,93.2％yield)。MS Calcd:666.23.；MS Found:567.22([M+H-100]+).
步骤4：向100mL茄形瓶中加入4-(4-(2-(2-氯-6-(三氟甲基)吡啶-3-基)氨基)-2-氧乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(51-d,650mg,0.97mmol)和溶剂DCM(8mL)。然后向其中加TFA(1mL)，室温搅拌过夜。用饱和碳酸氢钠溶液将pH调至碱性，用二氯甲烷萃取(30mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得到N-[2-氯-6-(三氟甲基)吡啶-3-基]-2-[2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(51-e,500mg,0.88mmol,90.5％yield)MS Calcd:566.18.；MS Found:567.12([M+H]+).
步骤5：向50mL茄形瓶中加入N-[2-氯-6-(三氟甲基)吡啶-3-基]-2-[2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(51-e,70mg,0.12mmol)和溶剂2-甲基-2-丁醇(2mL)。然后向其中加1-环丁基-5-羟基吡唑-4-甲酸乙酯(17-b,104mg,0.49mmol)，DIPEA(48mg,0.37mmol)，180℃搅拌30分钟。向其中加入甲醇(1.5mL)，粗品通过Pre-HPLC纯化得到标题化合物：N-[2-氯-6-(三氟甲基)吡啶-3-基]-2-(6-{4-[(1-环丁基-5-羟基吡唑-4-基)羰基]哌嗪-1-基}-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)乙酰胺(51,20mg,0.03mmol,21.1％yield).MS Calcd:730.24；MS Found:731.28([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.57(s,1H),8.58(d,J＝8.4Hz,1H),7.98(d,J＝8.4Hz,1H),7.70(s,1H),7.09–6.65(m,1H),5.39(s,2H),4.84-4.75(m,1H),4.34(d,J＝12.4Hz,2H),4.27(q,J＝2.8Hz,2H),3.82(t,J＝5.6Hz,2H),3.52-3.45(m,3H),3.21-3.15(m,2H),3.05-2.99(m,3H),2.78(d,J＝11.2Hz,2H),2.55–2.51(m,2H),2.35–2.26(m,2H),1.83-1.75(m,2H),1.22(t,J＝7.2Hz,3H).
实施例35 2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(化合物52)的制备
步骤1：向25mL茄形瓶中加入2,5-二氯-4-(三氟甲基)苯胺(52-a，800mg, 3.48mmol)和溶剂二氯甲烷(10mL)。然后向其中加入氯乙酰氯(589mg,5.22mmol)和三乙胺(1056mg,10.43mmol)，室温搅拌2小时。向其中加入水(40mL)，用DCM萃取(30mL)，饱和食盐水(30mL×2)洗涤有机相两次，无水硫酸钠干燥，减压旋干得2-氯-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(52-b，1000mg,3.26mmol,93.8％yield).未纯化直接下一步。MS Calcd:304.94；MS Found:303.95([M-H]-).
步骤2：向25mL茄形瓶中加入2-氯-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(52-b，1000mg,3.26mmol)和溶剂丙酮(20mL)。然后向其中加入碘化钾(1624.8mg,9.79mmol)，加热到50℃搅拌2小时。反应液过滤，滤液减压旋干得粗产品，通过硅胶层析柱纯化(EA:PE＝10％)得2-碘-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(52-c，700mg,1.76mmol,53.9％yield)。MS Calcd:396.87；MS Found:395.9([M-H]-).
步骤3：向25mL茄形瓶中加入化合物4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-5，200mg,0.46mmol)和溶剂1,4-二氧六环(10mL)。然后向其中加入2-碘-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(52-c，184.9mg,0.46mmol)和DIPEA(178.3mg,1.38mmol)，加热到75℃搅拌过夜。向其中加入水(30mL)，用EA萃取(30mL)，饱和食盐水(30mL×2)洗涤有机相两次，无水硫酸钠干燥，减压旋干得粗产品，通过薄层硅胶板纯化(MeOH:DCM＝1:20)得4-(4-(2-((2,5-二氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(52-d，200mg,0.29mmol,61.9％yield)。MS Calcd:699.20；MS Found:700.3([M+H]+).
步骤4：向25mL茄形瓶中加入4-(4-(2-((2,5-二氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(52-d，100mg,0.14mmol)和溶剂二氯甲烷(50mL)。然后向其中加入HCl/1,4-二氧六环(2mL,4mmol/L)，室温搅拌1小时。向其中加入饱和碳酸氢钠溶液(40mL)，用DCM萃取(30mL)，饱和食盐水(30mL×2)洗涤有机相两次，无水硫酸钠干燥，减压旋干得N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基l)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(52-e，79mg,0.13mmol,92.2％yield)未纯化直接下一步。MS Calcd:599.14；MS Found:600.20([M+H]+).
步骤5：向25mL茄形瓶中加入N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基l)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(52-e，35mg,0.06mmol)和溶剂叔戊醇(2mL)。然后向其中加入1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b，50.4mg,0.24mmol)，加热到180℃搅拌 1小时后反应液直接浓缩，粗品通过Pre-HPLC纯化得到标题化合物：2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(52，18mg,0.02mmol,40.4％yield)。MS Calcd:763.20；MS Found:764.22([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.51(s,1H),8.30(s,1H),8.07(s,1H),7.71(s,1H),6.84(s,1H),5.37(s,2H),4.84-4.76(m,1H),4.34(d,J＝12.4Hz,2H),4.27(d,J＝3.2Hz,2H),3.82(t,J＝5.6Hz,2H),3.52–3.45(m,3H),3.22-3.19(m,2H),3.06-2.99(m,3H),2.79(d,J＝11.2Hz,2H),2.32–2.28(m,2H),1.84-1.77(m,2H),1.26-1.20(m,5H).
实施例36 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-(2,2,2-三氟乙基)-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物53)的制备
步骤1：将5-羟基-1-(2,2,2-三氟乙基)-1H-吡唑-4-甲酸乙酯(37-b,105mg,0.44mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-(2,2,2-三氟乙基)-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(53,10mg，0.01mmol，14.3％yield)。MS Calcd:757.20；MS Found:758.27([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),8.08(d,J＝8.4Hz,1H),7.98(d,J＝2.0Hz,1H),7.81–7.62(m,2H),7.02–6.76(m,1H),5.34(s,2H),4.87-4.80(m,2H),4.31–4.26(m,4H),3.82(t,J＝5.6Hz,2H),3.52-3.46(m,2H),3.21–3.15(m,2H),3.05-2.99(m,2H),2.78(d,J＝11.2Hz,2H),2.53-2.50(m,2H),1.22(t,J＝7.2Hz,3H).
实施例37 (R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物54)的制备
步骤1：将2-溴-5-乙基-4,7-二氢[1,2,4]三唑并[1,5-a]嘧啶-7-酮(int-3a,700mg,2.88mmol)和N-[5-氯-2-甲基-4-(三氟甲基)苯基]-2-碘代乙酰胺(38-c,1087.0mg,2.88mmol)加入到100mL圆底烧瓶中，依次加入二氧六环(20mL)和DIPEA(1.43mL,8.64mmol)，氮气保护下于80℃搅拌3小时。将反应液降至室温，有固体析出，过滤，EA洗涤，合并滤液，浓缩快速柱层析(DCM:MeOH＝5:1)得2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)乙酰胺(54-a，603mg,1.22mmol，42.5％yield)。
步骤2：将2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)乙酰胺(54-a，600mg,1.22mmol)和2-(3,6-二氢-2H-吡喃-4-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(383.8mg,1.83mmol)溶于1,4-二氧六环(8mL)和H2O(4mL)中，再依次加入Pd(dppf)Cl2(89mg,0.09mmol)，磷酸钾(860mg,3.65mmol)，置换三次氮气，80℃反应5.0小时。将反应液浓缩，向残渣中加入30mL乙酸乙酯，经硅藻土过滤，滤液浓缩，经柱层析纯化(DCM:MeOH＝4:1)得N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54-b，200mg，0.4mmol，33％yield)。MS Calcd:495.13；MS Found:496.10([M+H]+).
步骤3：将N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54-b，450mg,0.91mmol)和N-溴代丁二酰亚胺(192.7mg,1.09mmol)置于100mL茄形瓶中，加入10mL乙腈，随后升高温度至60℃反应1小时。将反应液倒入20mL水中，用EA萃取(3×20mL)，合并有机相，干燥浓缩，经柱层析纯化(DCM:MeOH＝6:1)得到2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)乙酰胺(54-c，300mg，0.52mmol，57％yield)。MS Calcd:575.04；MS Found:576.20([M+H]+).
步骤4：将2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)乙酰胺(54-c，250mg， 0.43mmol)和化合物2-甲基丙-2-基(2R)-2-甲基哌嗪-1-甲酸酯(871mg，4.35mmol)，AgBF4(101.3mg,0.52mmol)加入到干燥的DMSO(3mL)中，氮气置换1分钟，然后升高温度到120℃反应18h。将反应体系倒入水中(20mL)，然后用EA萃取(20mLx3)，合并有机相，干燥浓缩，剩余物经Prep-TLC纯化(DCM：MeOH＝20:1)得到(R)-4-(4-(2-((5-氯-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(54-d，127mg，0.18mmol，42％yield)。MS Calcd:693.27；MS Found:694.10([M+H]+).
步骤5：将(R)-4-(4-(2-((5-氯-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(54-d，127mg，0.18mmol)溶于8mL二氯甲烷溶液中，再向其中加入2mL三氟乙酸，室温下搅拌2小时。将反应液直接浓缩，向残渣中加入20mL饱和碳酸氢钠溶液，用二氯甲烷萃取(20mL×3)，合并有机相，干燥浓缩，得到粗品(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54-e，90mg,0.15mmol,82％yield)。MS Calcd:593.21；MS Found:594.27([M+H]+).
步骤6：将(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54-e，50mg,0.08mmol)和1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,88mg,0.42mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物:(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54，13mg，0.02mmol，20％yield)。MS Calcd:757.27；MS Found:758.37([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.13(s,1H),7.98(s,1H),7.78(s,1H),7.69(s,1H),6.84(t,J＝2.0Hz,1H),5.30(d,J＝2.7Hz,2H),4.80(p,J＝9.3,8.8Hz,2H),4.28(q,J＝2.8Hz,2H),3.83(t,J＝5.6Hz,2H),3.73-3.67(m,2H),3.54-3.49(m,2H),3.22-3.15(m,2H),2.99-2.94(m,2H),2.80(d,J＝10.0Hz,2H),2.38(s,3H),2.32–2.28(m,2H),1.84-1.76(m,2H),1.46-1.43(m,3H),1.27-1.23(m,5H).
实施例38 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物55)的制备
步骤1：将(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,63mg,0.10mmol)，DIPEA(15mg，0.10mmol)，1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,112mg，0.5mmol)加入到2-甲基-2-丁醇(1mL)中，油浴升温到180℃，搅拌30分钟，将反应体系溶解到甲醇中，直接prep-HPLC纯化，得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(55,19mg,0.025mmol,25.0％yield)。MS Calcd:757.27；MS Found:758.31([M+H]+)。1H NMR(400MHz,Chloroform-d)δ9.03(s,1H),8.53(d,J＝8.8Hz,1H),7.68(d,J＝2.0Hz,1H),7.63–7.50(m,2H),7.01(s,1H),5.14(s,2H),4.80-4.73(m,1H),4.41–4.39(m,2H),4.01–3.94(m,3H),3.83-3.78(m,1H),3.34-3.18(m,2H),2.85–2.67(m,4H),2.24–2.04(m,1H),2.12–2.07(m,4H),2.00–1.94(m,3H),1.76-1.70(m,3H),1.40(t,J＝7.2Hz,3H),1.29(d,J＝9.2Hz,3H).
实施例39 (R)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物56)的制备
步骤1：将2-溴-5-乙基-[1,2,4]三唑并[1,5-a]嘧啶-7(4H)-酮(int-3a,4g,16.5mmol)，N-(3-氟-2-甲基-4-(三氟甲基)苯基)-2-碘代乙酰胺(int-4,6g,16.5mmol)，DIPEA(6.4g,49.5mmol)加入到DMF(50mL)中，升高温度到80℃，搅拌2小时， 冷却到室温，将反应体系倒入水中，有固体析出，抽滤，滤饼用水洗涤(50mLx2)，收集固体，用乙酸乙酯和石油醚(1:3)混合溶剂打浆纯化得到2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-a,5.9g,12.4mmol，75％yield)。MS Calcd:475.03；MS Found:476.11([M+H]+)。
步骤2：将2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-a,5.9g,12.4mmol)，2-(3,6-二氢-2H-吡喃-4-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(2.6g,12.4mmol)，磷酸钾(7.9g,37.2mmol)，1,1-双(二苯基膦)二荗铁二氯化钯(906mg,1.24mmol)加入到二氧六环(40mL)和水(10mL)的混合溶剂中，氮气置换3分钟，升高温度到85℃，搅拌3小时，减压浓缩，剩余物倒入水中，将固体经过滤收集，然后用乙醇打浆纯化得到2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-b,4.8g,10.0mmol，81％yield)。MS Calcd:479.16；MS Found:480.21([M+H]+)。
步骤3：将2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-b,4.8g,10.0mmol)加入到DMF中(50mL)，然后加入NBS(1.78g,10.0mmol)，升高温度到60℃，搅拌3小时。然后将反应体系倒入水中，有固体析出，固体经过滤收集，然后用乙醇打浆纯化得到2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-c,3.4g,6.1mmol，61％)。MS Calcd:557.07；MS Found:558.07([M+H]+)。
步骤4：将2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-c,1g,1.8mmol)，(R)-2-甲基哌嗪-1-甲酸叔丁酯(3.6g，18mmol)，四氟硼酸银(351mg，1.8mmol)加入到干燥的DMSO(10mL)中，然后氮气保护下，升高温度到120℃，搅拌4小时，冷却到室温，加入饱和碳酸氢钠水溶液，然后用乙酸乙酯萃取(10mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，剩余物经柱层析纯化(EA：PE＝1:1)，得到(R)-4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(-2-(3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(56-d,810mg,1.20mmol，66％)。MS Calcd:677.29；MS Found:678.31([M+H]+)。
步骤5：将(R)-4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(-2-(3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(56-d,810mg,1.20mmol)加入到DCM中(10mL)，然后加入三氟乙酸(3mL)，在室温下搅拌4小时，减压浓缩，剩余物加入饱和碳酸氢钠水溶液，然后用DCM萃取(10mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液 经减压浓缩，得到(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-e,483mg,0.84mmol,70％)。MS Calcd:577.24；MS Found:578.28([M+H]+)。
步骤6：将(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-e,60mg,0.10mmol)，DIEA(15mg，0.10mmol)，1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a，100mg，0.5mmol)加入到2-甲基-2-丁醇(1mL)中，油浴升温到180℃，搅拌30分钟，经prep-HPLC纯化，得到标题化合物：(R)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56,21mg,0.029mmol,29.0％yield)。MS Calcd:727.29；MS Found:728.31([M+H]+)。
实施例40 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-5-乙基-2-(4-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物57)的制备
步骤1：向25mL茄形瓶中加入4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2a,100mg,0.15mmol)和溶剂DMSO(2mL)。然后向其中加入N-甲基哌嗪(45.1mg,0.45mmol)和乙酸钾(44.1mg,0.45mmol)，然后加热到120℃搅拌过夜。向其中加入水(40mL)，用EA萃取(30mL)，饱和食盐水(30mL×2)洗涤有机相两次，无水硫酸钠干燥，减压旋干得粗产品，经柱层析纯化(MeOH:DCM＝1:20)得4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基l-2-(4-甲基哌嗪-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基l))哌嗪-1-甲酸叔丁酯(57-a，45mg,0.07mmol,44％yield)。MS Calcd:681.28；MS Found:682.4([M+H]+).
步骤2：向25mL茄形瓶中加入4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧 代乙基)-5-乙基l-2-(4-甲基哌嗪-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基l))哌嗪-1-甲酸叔丁酯(57-a，45mg,0.07mmol,44％yield)和溶剂二氯甲烷(5mL)。然后向其中加入氯化氢-1,4-二氧六环(2mL,4mol/L)，室温搅拌1小时。向其中加入饱和碳酸氢钠溶液(20mL)，用DCM萃取(30mL)，饱和食盐水(30mL×2)洗涤有机相两次，无水硫酸钠干燥，减压旋干得粗品N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲基哌嗪-1-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(57-b，40mg,0.07mmol)，直接用于下一步。MS Calcd:581.22；MS Found:582.3([M+H]+).
步骤3：向25mL茄形瓶中加入N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲基哌嗪-1-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(57-b，40mg,0.07mmol)和溶剂叔戊醇(2mL)。然后向其中加入1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b，57.8mg,0.27mmol)和DIPEA(17.8mg,0.14mmol)，加热到180℃搅拌1小时。反应液直接浓缩，粗品通过Pre-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-5-乙基-2-(4-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(57，15mg,0.02mmol,28.3％yield)。MS Calcd:745.28；MS Found:746.4([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.35(s,1H),8.06(dd,J＝8.4,6.0Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.4,2.0Hz,1H),7.63(s,1H),5.23(s,2H),4.83-4.75(m,1H),4.34(d,J＝12.4Hz,2H),3.50–3.42(m,8H),3.17-3.10(m,1H),2.97(q,J＝7.2Hz,2H),2.72(d,J＝11.2Hz,2H),2.54-2.51(m,2H),2.42-2.39(m,4H),2.27-2.22(m,5H),1.82-1.75(m,2H),1.19(t,J＝7.2Hz,3H).
实施例41 N-(2-氯-6-(三氟甲基)吡啶-3-基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物58)制备
步骤1：向50mL茄形瓶中加入N-[2-氯-6-(三氟甲基)吡啶-3-基]-2-[2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(51-e,70mg,0.12mmol)和溶剂2-甲基-2-丁醇(2mL)。然后向其中加1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a,97mg,0.49mmol)，DIPEA(48mg,0.37mmol)，180℃搅拌30分钟。向其中加入甲醇(1.5mL)，粗品通过Pre-HPLC纯化得到标题化合物：N-(2-氯-6-(三氟甲基)吡啶-3-基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶 -4(7H)-基)乙酰胺(58,20mg,0.03mmol,21.9％yield)。MS Calcd:716.22；MS Found:717.20([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.60(s,1H),8.59(d,J＝8.4Hz,1H),7.98(d,J＝8.4Hz,1H),7.59(s,1H),6.84(q,J＝1.6Hz,1H),5.39(s,2H),4.34(d,J＝12.4Hz,2H),4.27(d,J＝3.2Hz,2H),3.82(t,J＝5.6Hz,2H),3.51–3.38(m,3H),3.16(t,J＝12.4Hz,2H),3.02(q,J＝7.2Hz,2H),2.77(d,J＝11.2Hz,2H),2.53–2.51(m,2H),1.22(t,J＝7.2Hz,3H),1.01–0.94(m,4H).
实施例42 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-8-甲氧基喹啉-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物59)的制备
步骤1：称取4-羟基-8-甲氧基喹啉-3-甲酸(59-a,23mg,0.09mmol)，N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,40mg,0.06mmol)到25mL单口瓶中，加入2-甲基-2-丁醇(2mL)和DIPEA(0.01mL,0.06mmol)，加完后升温到180℃反应2小时。反应液浓缩，残余物prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-8-甲氧基喹啉-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(59,2mg,0.002mmol,4.3％Yeild)。MS Calcd:766.22；MS Found:767.36([M+H]+)。
实施例43 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物60)的制备
步骤1：将4-羟基异噁唑-3-甲酸(int-7,26mg,0.20mmol)加入到DCM中(1mL)，然后加入1-氯-N,N,2-三甲基丙基-1-烯-1-胺(29mg，0.20mmol)，室温下搅拌2小时，然后加入到(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,60mg,0.10mmol)和DIPEA(64mg，0.50mmol)的二氯甲烷溶液中，并在 室温下搅拌过夜，然后加入水(3mL)，用DCM萃取(3mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，剩余物经Prep-HPLC纯化，得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(60,16mg,0.023mmol,23.1％yield)。MS Calcd:690.19；MS Found:691.21([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),9.84(s,1H),8.59(d,J＝2.0Hz,1H),8.08(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),6.86-6.84(m,1H),5.34(s,2H),3.82-3.78(m,0.5H),4.40(d,J＝12.8Hz,,0.5H),4.27(d,J＝2.8Hz,2H),3.89-3.82(m,3H),3.72-3.67(m,1H),3.54–3.44(m,2H),3.31–3.12(m,3H),2.96-2.68(m,3H),1.43(dd,J＝6.8,2.0Hz,3H),1.23(t,J＝7.2Hz,3H).
实施例44 (R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物61)的制备
步骤1：将4-羟基异噁唑-3-甲酸(int-7,26mg,0.20mmol)加入到DCM中(1mL)，然后加入1-氯-N,N,2-三甲基丙基-1-烯-1-胺(29mg，0.20mmol)，室温下搅拌2小时，然后加入到(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-e,60mg,0.10mmol)和DIPEA(64mg，0.50mmol)的二氯甲烷溶液中，并在室温下搅拌过夜，然后加入水(3mL)，用DCM萃取(3mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，剩余物经Prep-HPLC纯化，得到标题化合物：(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(61,11mg,0.016mmol,16％yield)。MS Calcd:688.24；MS Found:689.27([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.23(s,1H),9.79(s,1H),8.59(d,J＝1.6Hz,1H),7.63-7.57(m,2H),6.84(t,J＝2.0Hz,1H),5.28(s,2H),4.82–4.78(m,0.5H),4.39(d,J＝12.8Hz,0.5H),4.29-4.26(m,2H),3.88-3.81(m,3H),3.72-3.66(m,1H),3.53–3.44(m,2.5H),3.26–3.13(m,2H),2.97-2.90(m,1H),2.85–2.62(m,2.5H),2.26(d,J＝2.4Hz,3H),1.43(dd,J＝6.8,2.0Hz,3H),1.23(t,J＝7.2Hz,3H).
实施例45 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-5-乙基-2-吗啡啉-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物62)的制备
步骤1：向25mL茄形瓶中加入4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2a,200mg,0.30mmol)和溶剂DMSO(2mL)。然后向其中加入吗啡啉(78.8mg,0.91mmol)和乙酸钾(88.8mg,0.91mmol)，然后加热到120℃搅拌过夜。向其中加入水(40mL)，用EA萃取(30mL)，饱和食盐水(30mL×2)洗涤有机相两次，无水硫酸钠干燥，减压旋干得粗产品，通过色谱硅胶板纯化(MeOH:DCM＝1:20)得4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基l)-5-乙基-2-吗啡啉-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基l))哌嗪-1-甲酸叔丁酯(62-a，55mg,0.08mmol,27.2％yield)。MS Calcd:668.24；MS Found:613.25([M+H-56]+).
步骤2：向25mL茄形瓶中加入4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基l)-5-乙基-2-吗啡啉-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基l))哌嗪-1-甲酸叔丁酯(62-a，55mg,0.08mmol)和溶剂二氯甲烷(5mL)。然后向其中加入氯化氢-1,4-二氧六环溶液(2mL,4mol/L)，室温搅拌1小时。向其中加入饱和碳酸氢钠溶液(20mL)，用DCM萃取(30mL)，饱和食盐水(30mL×2)洗涤有机相两次，无水硫酸钠干燥，减压旋干得粗品N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-吗啡啉-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基l)乙酰胺(62-b，40mg,0.08mmol)粗产品直接下一步。MS Calcd:568.19；MS Found:569.34([M+H]+).
步骤3：向25mL茄形瓶中加入N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-吗啡啉-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基l)乙酰胺(62-b，40mg,0.08mmol)和溶剂叔戊醇(2mL)。然后向其中加入1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b，66.5mg,0.32mmol)和DIPEA(20.4mg,0.16mmol)，加热到180℃搅 拌1小时，反应液直接浓缩，粗品通过Pre-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-5-乙基-2-吗啡啉-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(62，10mg,0.01mmol,16.7％yield)。MS Calcd:732.25；MS Found:733.3([M+H]+).1H NMR(400MHz,DMSO-d6+重水交换)δ8.02(d,J＝8.4Hz,1H),7.95(d,J＝2.0Hz,1H),7.73(dd,J＝8.4,2.0Hz,1H),7.66(s,1H),5.21(s,2H),4.81-4.72(m,1H),4.29(d,J＝12.4Hz,2H),3.69-3.64(m,2H),3.46–3.37(m,7H),3.16-3.10(m,2H),2.97-2.91(m,2H),2.72(d,J＝11.2Hz,2H),2.49–2.43(m,3H),2.27(s,2H),1.81-1.73(m,2H),1.18(t,J＝7.2Hz,3H).
实施例46 N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物63)的制备
步骤1：向10mL茄形瓶中加入4-羟基异噁唑-3-甲酸(int-7,12.9mg,0.10mmol)和溶剂DCM(3mL)。然后向其中加入1-氯-N,N,2-三甲基丙烯胺(10mg,0.08mmol)，室温搅拌1小时后，加入N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基l)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(52-e，30mg,0.05mmol)和DIPEA(19.4mg,0.15mmol)，继续室温搅拌2小时，向其中加入水(20mL)，用DCM萃取(20mL)，饱和食盐水洗涤有机相(20mL×2)，无水硫酸钠干燥，减压旋干得粗品，粗品通过Pre-HPLC纯化得到标题化合物：N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(63,7mg,0.01mmol,17.7％yield).MS Calcd:710.14；MS Found:711.3([M+H]+).1H NMR(400MHz,Methanol-d4)δ11.32(s,1H),10.59(s,1H),9.42(s,1H),9.11(s,1H),8.87(s,1H),7.65(s,1H),6.16(s,2H),5.35(d,J＝12.4Hz,1H),5.08(q,J＝2.8Hz,2H),4.63(t,J＝5.2Hz,2H),4.44(d,J＝12.4Hz,1H),4.33–4.26(m,2H),4.18–4.10(m,1H),3.88-3.80(m,3H),3.65(d,J＝11.6Hz,1H),3.55(d,J＝11.6Hz,1H),3.35-3.31(m,2H),2.02(t,J＝7.2Hz,3H).
实施例47 N-(2-氯-4-(三氟甲基)苯基)-2-(6-((R)-4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物64)的制备
步骤1：向250mL茄形瓶中加入化合物N-[2-氯-4-(三氟甲基)苯基]-2-碘代乙酰胺(8.2g,22.62mmol)，2-溴-5-乙基-4,7-二氢[1,2,4]三唑并[1,5-a]嘧啶-7-酮(int-3a,5.0g,20.57mmol)和溶剂1,4-二氧六环(120mL)。然后向其中加DIPEA(8.0g,61.70mmol)，80℃搅拌4小时。冷却至室温后过滤，固体用乙酸乙酯洗涤(50mL)，再用DCM溶解固体(300mL)，用水洗涤两次，有机相用无水硫酸钠干燥，减压浓缩得到得2-(2-溴-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)-N-[2-氯-4-(三氟甲基)苯基]乙酰胺(64-a,5.7g,11.91mmol,57.9％yield)MS Calcd:478.65.；MS Found:479.94([M+H]+).
步骤2：向100mL茄形瓶中加入2-(2-溴-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)-N-[2-氯-4-(三氟甲基)苯基]乙酰胺(64-a,800mg,1.67mmol)，2-(4-甲氧基环己-1-烯基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(400mg,1.67mmol)和溶剂1,4-二氧六环(6mL)，H2O(2mL)。然后向其中加磷酸钾(960mg，4.18mmol)，Pd(dppf)Cl2(120mg,1.67mmol)，90℃搅拌过夜。加入50mL水，用乙酸乙酯萃取萃取(50mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得到粗品，粗品通过柱层析纯化(PE:EA＝1:3)，得N-[2-氯-4-(三氟甲基)苯基]-2-[5-乙基-2-(4-甲氧基环己-1-烯基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(64-b,720mg,1.41mmol,84.5％yield)MS Calcd:509.91.；MS Found:510.12([M+H]+).
步骤3：向100mL茄形瓶中加入N-[2-氯-4-(三氟甲基)苯基]-2-[5-乙基-2-(4-甲氧基环己-1-烯基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(64-b,650mg,1.27mmol)和乙腈(10mL)。然后向其中加NBS(250mg，1.40mmol)，60℃搅拌1小时。将溶剂旋干，加入20mL水，用乙酸乙酯萃取(20mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得到粗品，粗品通过柱层析纯化(PE:EA＝1:1)，得2-[6-溴-5-乙基-2-(4-甲氧基环己-1-烯基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]-N-[2-氯-4-(三氟甲基)苯基]乙酰胺(64-c,500mg,0.87mmol,67.9％yield)MS Calcd:587.05.；MS Found:588.04([M+H]+).
步骤4：向100mL茄形瓶中加入2-[6-溴-5-乙基-2-(4-甲氧基环己-1-烯基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]-N-[2-氯-4-(三氟甲基)苯基]乙酰胺(64-c,500mg, 0.85mmol)，2-甲基丙-2-基(2R)-2-甲基哌嗪-1-甲酸酯(1700mg,8.49mmol)和溶剂DMSO(10mL)。然后向其中加四氟硼酸银(198mg，1.02mmol)，120℃搅拌24小时。加入30mL水，用乙酸乙酯萃取(30mL×3)，饱和食盐水洗涤有机相4次，无水硫酸钠干燥，减压旋干得到粗品，粗品通过柱层析纯化(PE:EA＝1:3)，得2-甲基丙-2-基(2R)-4-[4-(2-{[2-氯-4-(三氟甲基)苯基]氨基}-2-氧代乙基)-5-乙基-2-(4-甲氧基环己-1-烯基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-6-基]-2-甲基哌嗪-1-甲酸酯(64-d,340mg,0.48mmol,56.5％yield)MS Calcd:707.28.；MS Found:608.24([M+H-100]+).
步骤5：向100mL茄形瓶中加入2-甲基丙-2-基(2R)-4-[4-(2-{[2-氯-4-(三氟甲基)苯基]氨基}-2-氧代乙基)-5-乙基-2-(4-甲氧基环己-1-烯基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-6-基]-2-甲基哌嗪-1-甲酸酯(64-d,340mg,0.48mmol)和溶剂DCM(3mL)。然后向其中加TFA(0.5mL)，室温搅拌3小时。加入饱和碳酸氢钠溶液将pH调至碱性，用二氯甲烷萃取(30mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得粗品N-[2-氯-4-(三氟甲基)苯基]-2-[5-乙基-2-(4-甲氧基环己-1-烯基)-6-[(3R)-3-甲基哌嗪-1-基]-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(64-e,150mg,0.25mmol,51.4％yield)MS Calcd:607.23；MS Found:608.20([M+H]+).
步骤6：向50mL茄形瓶中加入N-[2-氯-4-(三氟甲基)苯基]-2-[5-乙基-2-(4-甲氧基环己-1-烯基)-6-[(3R)-3-甲基哌嗪-1-基]-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(64-e,60mg,0.10mmol)和溶剂2-甲基-2-丁醇(1.5mL)。然后向其中加1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b，83mg,0.39mmol)，DIPEA(39mg,0.30mmol)，180℃搅拌30分钟。向其中加入甲醇(1.5mL)，粗品通过Pre-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-((R)-4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(64,20mg,0.03mmol,26.2％yield，非对映体混合物)。MS Calcd:771.29；MS Found:772.28([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.09(d,J＝8.8Hz,1H),7.99(d,J＝2.0Hz,1H),7.75–7.71(m,2H),6.77-6.75(m,1H),5.34(s,2H),4.84-4.76(m,1H),3.72-3.68(m,1H),3.56-3.33(m,5H),3.29(s,3H),3.19-3.11(m,1H),2.99-2.92(m,1H),2.79(d,J＝10.0Hz,1H),2.67–2.08(m,7H),2.35-2.26(m,2H),2.20-2.13(m,1H),1.99-1.93(m,1H),1.84-1.74(m,2H),1.72-1.63(m,1H),1.44(s,3H),1.24(t,J＝7.2Hz,3H).
实施例48 N-[2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-((R)-4-((4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)乙酰胺(化合物65)的制备
步骤1：向50mL茄形瓶中加入化合物4-羟基异噁唑-3-甲酸(int-7,31mg,0.24mmol)和溶剂DCM(1mL)。然后向其中加(1-氯-2-甲基丙-1-烯基)二甲胺(33mg,0.25mmol)，室温搅拌3小时。再向其中加入N-[2-氯-4-(三氟甲基)苯基]-2-[5-乙基-2-(4-甲氧基环己-1-烯基)-6-[(3R)-3-甲基哌嗪-1-基]-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(64-e,50mg,0.08mmol)和DIPEA(103mg，0.8mmol)，室温搅拌2小时，将溶剂旋干，向其中加入甲醇(1.5mL)，粗品通过Pre-HPLC纯化得到标题化合物：N-[2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-((R)-4-((4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)乙酰胺(65,20mg,0.03mmol,34.4％yield，非对映体混合物).MS Calcd:718.22；MS Found:719.19([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.34(s,1H),9.88(s,1H),8.60(d,J＝2.0Hz,1H),8.09(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),6.76(t,J＝4.0Hz,1H),5.34(s,2H),3.91-3.84(m,1H),3.72-6.67(m,1H),3.55–3.45(m,3H),3.31-3.13(m,4H),2.96–2.83(m,2H),2.71-2.43(m,5H),2.20–2.14(m,1H),1.99–1.94(m,1H),1.74–1.63(m,1H),1.44(dd,J＝6.8,2.0Hz,3H),1.23(t,J＝7.2Hz,3H).
实施例49 N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-((R)-4-(5-羟基-1-甲基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物66)的制备
步骤1：向50mL茄形瓶中加入5-羟基-1-甲基-1H-吡唑-4-甲酸(3-c,12mg,0.08mmol)和溶剂DCM(1mL)。然后向其中加(1-氯-2-甲基丙-1-烯基)二甲胺(12mg,0.09mmol)，室温搅拌3小时。再向其中加入N-[2-氯-4-(三氟甲基)苯基]-2-[5-乙基-2-(4-甲氧基环己-1-烯基)-6-[(3R)-3-甲基哌嗪-1-基]-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(64-e,40mg,0.07mmol)和DIPEA(27mg，0.21mmol)，室温搅拌2小时，将溶剂旋干，向其中加入甲醇(1.5mL)，粗品通过prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-((R)-4-(5-羟基-1-甲基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并 [1,5-a]嘧啶-4(7H)-基)乙酰胺(66,6mg,0.01mmol,11.4％yield，非对映体混合物)。MS Calcd:731.26；MS Found:732.28([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.46(brs,2H),8.08(d,J＝8.8Hz,1H),7.97(d,J＝2.0Hz,1H),7.89(s,1H),7.73(dd,J＝8.8,2.0Hz,1H),6.75(d,J＝4.0Hz,1H),5.33(s,2H),3.72-3.65(m,4H),3.56-3.48(m,4H),3.29(s,3H),3.19-3.10(m,1H),2.97-2.90(m,1H),2.74–2.42(m,6H),2.20-2.12(m,1H),1.98–1.93(m,1H),1.73-1.63(m,1H),1.40(brs,3H),1.23(t,J＝7.2Hz,3H).
实施例50 N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物67)的制备
步骤1：将4-羟基异噁唑-3-甲酸(int-7,26.1mg,0.20mmol)和1-氯-N,N,2-三甲基丙烯胺(29.7mg,0.22mmol)分别加入到3mL二氯甲烷溶液中，室温下搅拌3小时，然后加入N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(20-b，60mg,0.10mmol)、DIEA(0.05mL,0.3mmol)，继续升温至40℃搅拌5h。向反应液中加入10mL饱和碳酸氢钠溶液，DCM(10mL×2)萃取，合并有机相，减压浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(67，25mg，0.04mmol，35％yield)。MS Calcd:704.21；MS Found:705.29([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),9.83(s,1H),8.61(s,1H),8.08(d,J＝8.4Hz,1H),7.98(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),6.75(t,J＝4.0Hz,1H),5.33(s,2H),4.53(d,J＝12.4Hz,1H),3.62(d,J＝12.4Hz,1H),3.54-3.44(m,4H),3.30(s,3H),3.05-2.97(m,3H),2.83(d,J＝11.2Hz,1H),2.73(d,J＝11.2Hz,1H),2.62-2.42(m,3H),2.20–2.13(m,1H),1.99–1.95(m,1H),1.72–1.64(m,1H),1.21(t,J＝7.2Hz,3H).
实施例51 (R)-N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物68)的制备
步骤1：将2-碘-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(52-c，420mg，1.06mmol)置于100mL茄形瓶中，加入1,4-二氧六环溶液10mL。再向其中依次加入2-溴-5-乙基-[1,2,4]三唑并[1,5-a]嘧啶-7(4H)-酮(int-3a,250mg,1.03mmol)和DIPEA(0.5mL,3.09mmol)。75℃下搅拌5小时。将反应液浓缩，经柱层析纯化(PE:EA＝2:1)得2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(68-a，300mg，0.58mmol，57％yield)MS Calcd:512.94；MS Found:513.94([M+H]+).
步骤2：将2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(68-a，300mg，0.58mmol)和2-(3,6-二氢-2H-吡喃-4-基)-4,4,5,5-四甲基-1,3,2-二氧杂硼烷(184.3mg,0.88mmol)溶于1,4-二氧六环(8mL)和H2O(4mL)中，再依次加入Pd(dppf)Cl2(42.7mg,0.04mmol)，磷酸钾(403mg,1.75mmol)，置换三次氮气，80℃反应5.0小时。将反应液浓缩，向残余物中加入30mL乙酸乙酯，经硅藻土过滤，滤液浓缩，经柱层析纯化(DCM:MeOH＝5:1)得N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(68-b，220mg，0.33mmol，73％yield)MS Calcd:515.07；MS Found:516.28([M+H]+).
步骤3：将N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(68-b，220mg，0.33mmol)和N-溴代丁二酰亚胺(69.9mg,0.40mmol)置于100mL茄形瓶中，加入10mL乙腈，随后升高温度至60℃反应1小时。将反应液倒入20mL水中，用EA萃取(3×20mL)，合并有机相，干燥浓缩，经柱层析纯化(DCM:MeOH＝6:1)得到2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(68-c，50mg,0.08mmol,25％yield)MS Calcd:594.98；MS Found:595.92([M+H]+).
步骤4：将2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2,5-二氯-4-(三氟甲基)苯基)乙酰胺(68-c，50mg, 0.08mmol)和(R)-1-N-Boc-2-甲基哌嗪(168mg,0.84mmol)，AgBF4(16.4mg,0.08mmol)加入到干燥的DMSO(1mL)中，氮气置换1分钟，然后升高温度到120℃反应18h。将反应体系倒入水中(20mL)，然后用EA萃取(20mLx3)，合并有机相，干燥浓缩，剩余物经Prep-TLC纯化(DCM：MeOH＝20:1)得(R)-4-(4-(2-((2,5-二氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(68-d，25mg，0.03mmol，41％yield)MS Calcd:713.21；MS Found:714.21([M+H]+).
步骤5：将(R)-4-(4-(2-((2,5-二氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(68-d，25mg，0.03mmol)溶于4mL二氯甲烷溶液中，再向其中加入1mL三氟乙酸，室温下搅拌2小时。将反应液直接浓缩，向残渣中加入20mL饱和碳酸氢钠溶液，用二氯甲烷萃取(20mL×3)，合并有机相，干燥浓缩，得到粗品(R)-N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(68-e，20mg，0.03mmol，93％yield)MS Calcd:613.16；MS Found:614.16([M+H]+).
步骤6：将4-羟基异噁唑-3-甲酸(int-7,12.6mg,0.10mmol)和1-氯-N,N,2-三甲基丙烯胺(13.0mg,0.10mmol)分别加入到3mL二氯甲烷溶液中，室温下搅拌3小时，然后加入(R)-N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(68-e，20mg，0.03mmol)、DIPEA(0.02mL,0.1mmol)，继续升温至40℃搅拌5h。向反应液中加入10mL饱和碳酸氢钠溶液，DCM(10mL×2)萃取，合并有机相，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(68，3mg,0.01mmol,12％yield)。MS Calcd:724.15；MS Found:725.24([M+H]+).
1H NMR(400MHz,DMSO-d6)δ10.52(s,1H),9.80(s,1H),8.60(d,J＝1.6Hz,1H),8.30(s,1H),8.04(s,1H),6.85-6.83(m,1H),5.34(s,2H),4.81-4.78(m,0.5H),4.40(d,J＝12.8Hz,0.5H),4.27(d,J＝2.8Hz,2H),3.88-3.80(m,3H),3.71–3.67(m,1H),3.54–3.44(m,2H),3.27–3.11(m,2H),2.96–2.83(m,2H),2.73–2.63(m,2H),1.43(dd,J＝6.8,2.0Hz,3H),1.24(t,J＝7.2Hz,3H).
实施例52 (R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物69)的制备
步骤1：将1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a,82mg,0.42mmol)和(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54-e，50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(69,15mg,0.02mmol,24％yield).MS Calcd:743.26；MS Found:744.31([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.13(s,1H),7.98(s,1H),7.78(s,1H),7.57(s,1H),6.84(t,J＝2.0Hz,1H),5.30(s,2H),4.68-4.61(m,1H),4.29-4.20(m,3H),3.82(t,J＝5.2Hz,2H),3.72-3.68(m,1H),3.53-3.48(m,1H),3.42–3.14(m,5H),2.99-2.94(m,1H),2.78(d,J＝10.0Hz,1H),2.765(d,J＝11.6Hz,1H),2.38(s,3H),1.43(d,J＝6.4Hz,3H),1.24(t,J＝7.2Hz,3H),1.00–092(m,4H).
实施例53 (R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物70)的制备
步骤1：将4-羟基异噁唑-3-甲酸(int-7,32.6mg,0.25mmol)和1-氯-N,N,2-三甲基丙烯胺(33.7mg,0.25mmol)分别加入到3mL二氯甲烷溶液中，室温下搅拌3小时，然后加入(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54-e，50mg，0.08mmol)、DIPEA(0.04mL,0.2mmol)，继续升温至40℃搅拌5h。向反应液中加入10mL饱和碳酸氢钠溶液，DCM(10mL×2)萃取，合并有机相，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌 嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(70，16mg，0.02mmol，27％yield)。MS Calcd:704.21；MS Found:705.26([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.12(s,1H),9.79(s,1H),8.60(d,J＝2.0Hz,1H),7.98(s,1H),7.77(s,1H),6.85–6.83(m,1H),5.29(s,2H),4.82–4.78(m,0.5H),4.40(d,J＝12.8Hz,0.5H),4.27(d,J＝2.8Hz,2H),3.89–3.81(m,3H),3.72-3.67(m,1H),3.54–3.4(m,2H),3.30-3.13(m,2H),2.97–2.83(m,1.5H),2.73–2.63(m,1.5H),2.54–2.51(m,1H),2.38(s,3H),1.43(dd,J＝6.8,2.0Hz,3H),1.23(t,J＝7.2Hz,3H).
实施例54 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物71)的制备
步骤1：称取4-羟基异噁唑-3-甲酸(Int-7,25mg,0.19mmol)，HATU(97.5mg,0.26mmol)到25mL单口瓶中，加入DCM(2mL)和DIPEA(0.06mL,0.38mmol)，加完后室温搅拌5分钟后加入2-(2-(3,6-二氢吡喃-4-基)-5-乙基-7-氧代-6-哌嗪-1-基-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)基)-N-(2-甲基-4-三氟甲基苯基)乙酰胺(1-c,70mg,0.13mmol)，加完后室温反应1小时。将反应液加入10mL饱和氯化铵溶液搅拌，然后分液，浓缩有机相，残余物prep-HPLC纯化得到标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(71,22mg,0.03mmol,94％Yield)。MS Calcd:656.23；MS Found:657.32([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.02(s,1H),9.75(s,1H),8.60(s,1H),7.74(t,J＝7.2Hz,1H),7.64(d,J＝2.0Hz,1H),7.56–7.53(m,1H),6.85(s,1H),5.27(s,2H),4.53(d,J＝12.4Hz,1H),4.28(q,J＝2.8Hz,2H),3.83(t,J＝5.6Hz,2H),3.63(d,J＝12.4Hz,1H),3.52–3.45(m,2H),3.37–3.32(m,1H),3.07-2.99(m,4H),2.84(d,J＝11.2Hz,1H),2.73(d,J＝11.2Hz,1H),2.55-2.48(m,1H),2.37(s,3H),1.22(t,J＝7.2Hz,3H).
实施例55 (R)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物72)的制备
步骤1：将2-溴-5-乙基-[1,2,4]三唑并[1,5-a]嘧啶-7(4H)-酮(int-3a,2.0g,8.23mmol)、N-(5-氟-2-甲基-4-(三氟甲基)苯基)-2-碘乙酰胺(3.0g,8.23mmol)和DIPEA(3.2g,24.68mmol)分别加入到60mL二氧六环溶液中，于80℃条件下搅拌4小时。将反应液减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-a,3.6g,7.56mmol,91.9％yield)。MS Calcd:475.03；MS Found:476.05([M+H]+).
步骤2：将2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-a,3.6g,7.56mmol)、3,6-二氢-2H-吡喃-4-硼酸频哪醇酯(1.58g,7.5mmol)、Pd(dppf)Cl2·CH2Cl2(1.07g,1.32mmol)和磷酸钾(4.77g,22.5mmol)依次加入40mL二氧六环/水(4:1)混合溶液中，抽真空氮气置换3次，升温至80℃搅拌2h。将反应液完全浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-b,1.5g,3.2mmol,41.6％yield)。MS Calcd:479.16；MS Found:480.19([M+H]+).
步骤3：将2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-b,1.34g,2.8mmol)溶于DMF(10mL)溶液中，然后缓慢加入NBS(0.55g,3.07mmol)，升温至60℃搅拌3h。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA)分离纯化得2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-c,1.3g,2.33mmol,83.3％yield)。MS Calcd:557.07；MS Found:558.15([M+H]+).
步骤4：将2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-c,1.3g,2.33mmol)、(R)-1-N-Boc-2-甲基哌嗪(5.02g,25.08mmol)和四氟硼酸银(0.58g,3.01 mmol)依次加入DMSO(10mL)溶液中，抽真空，氮气置换3次，升温至120℃搅拌过夜。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析分离纯化得(R)-4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(-2-(5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1，2，4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(72-d,1.2g,1.77mmol,75.9％yield)。MS Calcd:677.29；MS Found:678.32([M+H]+).
步骤5：将(R)-4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(-2-(5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1，2，4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(72-d,1.2g,1.77mmol加入盐酸-二氧六环溶液中(20mL)，置于室温搅拌过夜。将反应液完全浓缩，向残余物中加入饱和碳酸氢钠溶液，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得粗品(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-e,0.6g,1.03mmol,58.8％yield)。MS Calcd:577.24；MS Found:578.06([M+H]+).
步骤6：将1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a,102mg,0.52mmol)、DIPEA(40mg,0.31mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-e,50mg,0.09mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72,20mg,0.03mmol,25.1％yield)。MS Calcd:727.29；MS Found:728.37([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.11(s,1H),7.81(d,J＝12.4Hz,1H),7.69(d,J＝8.4Hz,1H),7.59(s,1H),6.85–6.83(m,1H),5.32(s,2H),4.67-4.58(m,1H),4.27-4.13(m,3H),3.82(t,J＝5.2Hz,2H),3.70(dd,J＝11.8,3.6Hz,1H),3.54–3.39(m,3H),3.21-3.14(m,1H),2.99-2.91(m,1H),2.79(d,J＝10.0Hz,1H),2.66(d,J＝11.6Hz,1H),2.54-2.49(m,2H),2.38(s,3H),1.43(d,J＝6.4Hz,3H),1.24(t,J＝7.2Hz,3H),1.03–0.93(m,4H).
实施例56 (R)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物73)的制备
步骤1：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b，105mg,0.5mmol)、DIPEA(40mg,0.31mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-e,60mg,0.1mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物送prep-HPLC纯化得标题化合物：(R)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(73,18mg,0.02mmol,23.7％yield)。MS Calcd:741.30；MS Found:742.34([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.09(s,1H),7.81(d,J＝12.8Hz,1H),7.71–7.67(m,2H),6.85–6.83(m,1H),5.32(s,2H),4.80(p,J＝8.4Hz,1H),4.65–4.59(m,1H),4.28-4.18(m,3H),3.82(t,J＝5.2Hz,2H),3.71(dd,J＝12.0,3.6Hz,1H),3.54–3.49(m,1H),3.21-3.14(m,1H),2.98-2.92(m,1H),2.79(d,J＝10.0Hz,1H),2.69–2.64(m,1H),2.56–2.49(m,3H),2.38(s,3H),2.37-2.27(m,2H),1.84-1.76(m,2H),1.44(d,J＝6.4Hz,3H),1.32-1.22(m,5H).
实施例57 (R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物74)的制备
步骤1：将4-羟基异噁唑-3-甲酸(int-7,40mg,0.31mmol)和1-氯-N,N,2-三甲基丙烯胺(44mg,0.33mmol)分别加入到DCM溶液中(2mL)，室温下搅拌3小时，然后加入(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-e,60mg,0.1mmol)、DIPEA(80mg,0.62mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(74，15mg,0.02mmol,21.7％yield)。MS Calcd:688.24；MS Found:689.24([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.11(s,1H),9.90(s,1H),8.60(d,J＝1.6Hz,1H),7.81(d,J＝12.8Hz,1H),7.69(d,J＝8.4Hz,1H),6.85–6.83(m,1H),5.32(s,2H),4.82-4.77(m,0.5H),4.40(d,J＝12.8Hz,0.5H),4.27(d,J＝2.8Hz,2H),3.89–3.81(m,3H),3.72-3.67(m,1H),3.54–3.44(m,2H),3.31-3.12(m,3H),2.96-2.83(m,1.5H),2.72-2.63(m,1.5H),2.37 (s,3H),1.43(dd,J＝6.4,2.0Hz,3H),1.23(t,J＝7.2Hz,3H).
实施例58 N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物75)的制备
步骤1：向250mL茄形瓶中加入3-氟-4-(三氟甲基)苯胺(75-a,10g,55.83mmol)，DCM(50mL)，依次加入BF3OEt2(7.9g,55.83mmol)，NCS(8.2g,61.42mmol)，加毕室温下搅拌反应3小时。减压浓缩反应液，剩余物经快速柱层析(PE:EA＝95:5)得2-氯-5-氟-4-(三氟甲基)苯胺(75-b,8.1g,37.92mmol,67.9％).MS Calcd:213.00；MS Found:212.12([M-H]-).
步骤2：向250mL茄形瓶中加入2-氯-5-氟-4-(三氟甲基)苯胺(75-b,8.1g,37.92mmol)，DCM(50mL)，然后加入三乙胺(10.51mL,75.84mmol)，在冰浴下缓慢加入氯乙酰氯(3.04mL,37.92mmol)的DCM(10mL)溶液，自然恢复室温反应过夜。浓缩反应液，剩余物快速柱层析(PE:EA＝92:8)，得2-氯-N-[2-氯-5-氟-4-(三氟甲基)苯基]乙酰胺(75-c，4.5g,15.52mmol,40.9％)。MS Calcd:288.97；MS Found:288.18([M+H]-).
步骤3：向250mL茄形瓶中加入2-氯-N-[2-氯-5-氟-4-(三氟甲基)苯基]乙酰胺(75-c，4.6g,27.59mmol)，丙酮(30mL)，然后50℃反应3小时。减压浓缩反应液，剩余物经快速柱层析(PE:DCM＝70:30)得到N-[2-氯-5-氟-4-(三氟甲基)苯基]-2-碘代乙酰胺(75-d，2.23g,5.85mmol,42.1％)。MS Calcd:380.90；MS Found:379.93([M-H]-).
步骤4：向50mL茄形瓶中加入4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-5,205.2mg,0.48mmol)，二氧六环(10mL)、N-[2-氯-5-氟-4-(三氟甲基)苯基]-2-碘代乙酰胺(75-d，200mg,0.55mmol)和DIPEA(0.25mL,1.50mmol)，氮气保护下80℃反应2hr。浓缩反应液，剩余物经快速柱层析(DCM：MeOH＝20:1)，得到4-(4-(2-(2-氯-5-氟-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代- 4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(75-e，292mg,0.43mmol,89.6％)。MS Calcd:683.22；MS Found:684.36([M+H]+).
步骤5：向50mL茄形瓶中加入4-(4-(2-(2-氯-5-氟-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(75-e，290mg,0.42mmol)和溶剂DCM(10mL)，加入TFA(3mL)，室温下搅拌2小时。向反应液中加入饱和碳酸氢钠溶液，调至pH＝8，DCM(3*50mL)萃取，合并有机相，得N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(75-f，221mg,0.38mmol,89.3％)。MS Calcd:583.17；MS Found:584.2([M+H]+).
步骤6：向25mL茄形瓶中加入N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(75-f，60mg,0.10mmol)和1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,21.6mg,0.10mmol)，依次加入2-甲基-2-丁醇(1mL)溶液，DIPEA(0.04mL)，抽真空氮气置换3次，180℃微波反应30min。减压旋干得粗品，粗品通过prep-HPLC纯化得到标题化合物：N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡唑-4基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(75,16mg,0.02mmol,20.8％yield).MS Calcd:747.23；MS Found:748.28([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.55(s,1H),8.13(d,J＝12.8Hz,1H),8.03(d,J＝7.2Hz,1H),7.56(s,1H),6.85–6.83(m,1H),5.39(s,2H),4.82-4.73(m,1H),4.37(d,J＝12.4Hz,2H),4.27(q,J＝2.8Hz,2H),3.82(t,J＝5.6Hz,2H),3.51-3.44(m,2H),3.12–2.98(m,4H),2.73(d,J＝11.2Hz,2H),2.48–2.43(m,2H),2.27-2.19(m,2H),1.85–1.64(m,2H),1.26-1.19(m,5H).
实施例59 (R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物76)的制备
步骤1：将2-溴-5-乙基-[1,2,4]三唑并[1,5-a]嘧啶-7(4H)-酮(int-3a,600mg,2.47mmol)，N-[2-氯-5-氟-4-(三氟甲基)苯基]-2-碘代乙酰胺(75-d，986.9mg,2.71mmol)加入到100mL圆底烧瓶中，加入二氧六环(30mL)，DIPEA(1.23mL,7.40mmol)氮气保护下于80℃搅拌6h。浓缩反应液，剩余物快速柱层析(PE:EA＝1:5)得2-(2-溴-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)-N-[2-氯-5-氟-4-(三氟甲基)苯基]乙酰胺(76-a,1.04g,2.09mmol,84.8％)。MS Calcd:494.97,496.97；MS Found:495.99,497.94([M+H]+).
步骤2：将2-(2-溴-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)-N-[2-氯-5-氟-4-(三氟甲基)苯基]乙酰胺(76-a,1.0g,2.01mmol)，3,6-二氢-2H-吡喃-4-硼酸频哪醇酯(0.6g,3.02mmol)，Pd(dppf)Cl2(0.1g,0.20mmol)，磷酸钾(1.4g,6.04mmol)加入到水(5mL)和二氧六环(20mL)混合溶剂中，升高温度到85℃，搅拌2小时。浓缩反应液，剩余物快速柱层析(PE:EA＝1:5)得N-[2-氯-5-氟-4-(三氟甲基)苯基]-2-[2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(76-b,508mg,1.02mmol,50.5％)。MS Calcd:499.10；MS Found:500.15([M+H]+).
步骤3：将N-[2-氯-5-氟-4-(三氟甲基)苯基]-2-[2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基]乙酰胺(76-b,0.45g,0.9mmol)溶于DMF(5mL)溶液中，然后缓慢加入NBS(0.18g,1.0mmol)，升温至60℃搅拌3h。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA％100)分离纯化得2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-5-氟-4-(三氟甲基)苯基)乙酰胺(76-c,0.45g,0.78mmol,86.3％yield)。MS Calcd:577.01；MS Found:578.07([M+H]+).
步骤4：将2-(6-溴-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-5-氟-4-(三氟甲基)苯基)乙酰胺(76-c,0.45g,0.78mmol)、(R)-1-N-Boc-2-甲基哌嗪(1.56g,7.78mmol)和四氟硼酸银(0.18g,0.93 mmol)依次加入DMSO(4mL)溶液中，抽真空，氮气置换3次，升温至120℃搅拌过夜。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA％100)分离纯化得(R)-4-(4-(2-(2-氯-5-氟-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(76-d,0.48g,0.69mmol,88.4％yield)。MS Calcd:697.24；MS Found:643.0([M-56+H]+).
步骤5：将(R)-4-(4-(2-(2-氯-5-氟-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(76-d,0.48g,0.69mmol)加入盐酸-二氧六环溶液中(20mL)，置于室温搅拌过夜。将反应液完全浓缩，向残余物中加入饱和碳酸氢钠溶液，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得(R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(76-e,0.3g,0.5mmol,73.0％yield)。MS Calcd:597.19；MS Found:598.16([M+H]+).
步骤6：将1-环丙基-5-羟基-1H-吡唑-4-甲酸乙酯(18-a,82mg,0.42mmol)、DIPEA(32mg,0.25mmol)和(R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(76-e,50mg,0.08mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物送prep-HPLC纯化得标题化合物：R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(6-(4-(1-环丙基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(75,15mg,0.02mmol,23.4％yield)。MS Calcd:747.23；MS Found:748.24([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.50(s,1H),8.14(d,J＝12.8Hz,1H),8.04(d,J＝7.2Hz,1H),7.59(s,1H),6.85-6.83(m,1H),5.39(s,2H),4.63(brs,1H),4.29-4.17(m,3H),3.82(t,J＝5.2Hz,2H),3.70(d,J＝11.2Hz,1H),3.54–3.40(m,2H),3.18–3.13(m,1H),2.98-2.91(m,1H),2.79(d,J＝10.0Hz,1H),2.66(d,J＝11.2Hz,1H),2.55-2.48(m,1H),1.43(d,J＝6.4Hz,3H),1.27-1.22(m,5H),1.02–0.93(m,4H).
实施例60 N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物77)的制备
步骤1：向25mL茄形瓶中加入4-羟基异噁唑-3-甲酸(int-7,33.2mg,0.26mmol)，(1-氯-2-甲基丙-1-烯基)二甲胺(35.3mg,0.27mmol)，DCM(2mL)，氮气保护下搅拌3小时，再加入DIPEA(0.14mL,0.86mmol)，N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(75-f，50mg,0.09mmol)，室温反应2hr。减压旋干得粗品，粗品通过prep-HPLC纯化得到标题化合物：N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(77,16.1mg,0.02mmol,24.4％yield).MS Calcd:694.17；MS Found:695.2([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.31(brs,1H),8.60(s,1H),8.14(d,J＝12.8Hz,1H),7.99(d,J＝7.2Hz,1H),6.84(t,J＝2.0Hz,1H),5.35(s,2H),4.54(d,J＝12.4Hz,1H),4.27(q,J＝2.8Hz,2H),3.82(t,J＝5.6Hz,2H),3.64(d,J＝12.4Hz,1H),3.52-3.46(m,3H),3.07–2.97(m,3H),2.84(d,J＝11.2Hz,1H),2.74(d,J＝11.2Hz,1H),2.54-2.48(m,2H),1.21(t,J＝7.2Hz,3H).
实施例61 (R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物78)的制备
步骤1：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,88mg,0.42mmol)、DIPEA(32mg,0.25mmol)和(R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(76-e,50mg,0.08mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物送prep-HPLC纯化得标题化合物：(R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(78,16mg,0.02mmol,24.3％yield)。MS Calcd: 761.25；MS Found:762.29([M+H]+).1H NMR(400MHz,DMSO-d6+重水交换)δ8.06(d,J＝12.4Hz,1H),7.99(d,J＝7.2Hz,1H),7.41(s,1H),6.80(s,1H),5.34(s,2H),4.80-4.73(m,1H),4.63(brs,1H),4.25–4.18(m,3H),3.79–3.76(m,2H),3.64-3.60(m,1H),3.48-3.42(m,1H),3.15-3.10(m,1H),2.94–2.88(m,1H),2.73(d,J＝9.2Hz,1H),2.61(d,J＝11.2Hz,1H),2.48–2.35(m,4H),2.16(brs,2H),1.73-1.67(m,2H),1.41(brs,4H),1.23-1.19(m,5H).
实施例62 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-(四氢-2H-吡啶-4-基)-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物79)的制备
步骤1：将乙氧基甲叉丙二酸二乙酯(3-a,0.7g,3.28mmol)、四氢-2H-吡喃-4-基肼盐酸盐(0.5g，3.28mmol)和碳酸钾(1.36g,9.83mmol)分别加入到水溶液中(10mL)，置于100℃条件下搅拌6小时。用5N HCl调至pH＝2，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得5-羟基-1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-甲酸乙酯(79-a,0.6g,2.5mmol,76.2％yield)。MS Calcd:240.11；MS Found:241.13([M+H]+).
步骤2：将5-羟基-1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-甲酸乙酯(79-a,106mg,0.44mmol)、DIPEA(34mg,0.27mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,50mg,0.09mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物送prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-(四氢-2H-吡啶-4-基)-1H-吡唑-4-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(79,12mg,0.02mmol,17.1％yield)。MS Calcd:759.25；MS Found:760.25([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.38(s,1H),8.08(d,J＝8.4Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.69(s,1H),6.86-6.84(m,1H),5.35(s,2H),4.42-4.34(m,4H),4.27(q,J＝2.8Hz,2H),4.00–3.95(m,2H),3.82(t,J＝5.6Hz,2H),3.52-3.45(m,4H),3.20-3.14(m,2H),3.03(q,J＝7.2Hz,2H),2.78(d,J＝11.2Hz,2H),2.52–2.48(m,2H),2.06-1.96(m,2H),1.80–1.75(m,2H),1.23(t,J＝7.2Hz,3H).
实施例63 (R)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2- (3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物80)的制备
步骤1：将1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,97mg,0.43mmol)、DIPEA(34mg,0.26mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-e,50mg,0.09mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(80,8mg,0.01mmol,11.1％yield)。MS Calcd:755.32；MS Found:756.34([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.10(s,1H),7.81(d,J＝12.8Hz,1H),7.70–7.67(m,2H),6.85–6.83(m,1H),5.32(s,2H),4.69–4.64(m,1H),4.28(q,J＝2.8Hz,2H),3.83(t,J＝5.2Hz,2H),3.73–3.69(m,1H),3.52(t,J＝11.2Hz,1H),3.20-3.15(m,1H),2.97-2.93(m,1H),2.81(d,J＝10.0Hz,1H),2.70–2.66(m,1H),2.52–2.48(m,2H),2.38(s,3H),2.03–1.78(m,7H),1.68–1.62(m,2H),1.45(brs,3H),1.26–1.23(m,5H),酰胺氢未出峰.
实施例64 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(cis-2,6-二甲基吗啉基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物81)的制备
步骤1：将4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2a,0.4g, 0.6mmol)、顺式2,6-二甲基吗啉(0.35g，3.02mmol)和乙酸钾(0.35g,3.62mmol)分别加入到DMSO溶液中(3mL)，升温至120℃条件下搅拌过夜。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:1)分离纯化得4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧乙基)-2-(cis-2,6-二甲基吗啉基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(81-a,0.17g,0.24mmol,40.4％yield)。MS Calcd:696.28；MS Found:641.19([M-56+H]+).
步骤2：将4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧乙基)-2-(cis-2,6-二甲基吗啉基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(81-a,0.17g,0.24mmol)加入盐酸-二氧六环溶液中(4mL)，置于室温搅拌过夜。将反应液完全浓缩，向残余物中加入饱和碳酸氢钠溶液，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得粗品N-(2-氯-4-(三氟甲基)苯基)-2-(2-(cis-2,6-二甲基吗啉基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(81-b,0.11g,0.18mmol,75.6％yield)。MS Calcd:596.22；MS Found:597.23([M+H]+).
步骤3：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,70mg,0.34mmol)、DIPEA(26mg,0.2mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(cis-2,6-二甲基吗啉基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(81-b,40mg,0.07mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(cis-2,6-二甲基吗啉基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(81,0.85mg,0.001mmol,1.6％yield)。MS Calcd:760.28；MS Found:759.23([M-H]-).
实施例65 N-[2-氯-4-(三氟甲基)苯基]-2-(5-乙基-6-{4-[(4-羟基异噁唑-3-基)酰基]哌嗪-1-基}-2-(1-甲基-1,2,3,6-四氢吡啶-4-基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)乙酰胺(化合物83)的制备
步骤1：向100mL茄形瓶中加入4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)- 2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2a,400mg,0.60mmol)，1-甲基-4-(4,4,5,5-四甲基-1,3,2-二氧杂硼烷-2-基)-1,2,3,6-四氢吡啶(202mg,0.91mmol)和溶剂1,4-二氧六环(8mL)和H2O(2mL)。然后向其中加磷酸钾(417mg，1.81mmol)，Pd(dppf)Cl2(44mg,0.06mmol)，90℃搅拌3小时。加入30mL水，用乙酸乙酯萃取(30mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得到粗品，粗品通过柱层析纯化(DCM:MeOH＝30:1)得4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-2-(1-甲基-1,2,3,6-四氢吡啶-4-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(83-a,300mg,0.44mmol,73.2％yield).MS Calcd:678.27.；MS Found:679.31([M+H]+).
步骤2：向100mL茄形瓶中加入4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-2-(1-甲基-1,2,3,6-四氢吡啶-4-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(83-a,300mg,0.44mmol)溶剂DCM(2mL)。然后向其中加TFA(0.5mL)，室温搅拌3小时。加入饱和碳酸氢钠溶液将pH调至碱性，用二氯甲烷萃取(20mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得粗品N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(1-甲基-1,2,3,6-四氢吡啶-4-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(83-b,200mg,0.35mmol,78.2％yield)MS Calcd:578.21；MS Found:579.41([M+H]+).
步骤3：向50mL茄形瓶中加入4-羟基异噁唑-3-甲酸(int-7,34mg,0.26mmol)和溶剂DCM(2mL)。然后向其中加(1-氯-2-甲基丙-1-烯基)二甲胺(36mg,0.27mmol)，室温搅拌3小时。再向其中加入N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(1-甲基-1,2,3,6-四氢吡啶-4-基)-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(83-b,50mg,0.09mmol)和DIPEA(101mg，0.86mmol)，室温搅拌3小时，将溶剂旋干，向其中加入甲醇(1.5mL)，残余物经prep-HPLC纯化得标题化合物：N-[2-氯-4-(三氟甲基)苯基]-2-(5-乙基-6-{4-[(4-羟基异噁唑-3-基)酰基]哌嗪-1-基}-2-(1-甲基-1,2,3,6-四氢吡啶-4-基)-7-氧代[1,2,4]三唑并[1,5-a]嘧啶-4-基)乙酰胺(83,10mg,0.01mmol,16.6％yield)MS Calcd:689.21；MS Found:690.29([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.40(s,1H),8.60(s,1H),8.29(s,1H),8.08(d,J＝8.4Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),6.79(t,J＝3.2Hz,1H),5.34(s,2H),4.54(d,J＝12.4Hz,1H),3.64(d,J＝12.4Hz,1H),3.52-3.45(m,2H),3.38-3.30(m,1H),3.04-2.98(m,4H),2.84(d,J＝11.2Hz,1H),2.74(d,J＝11.2Hz,1H),2.58-2.50(m,5H),2.31(s,3H),1.21(t,J＝7.2Hz,3H).
实施例66 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,4-二氢-2H-吡喃-6-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物84)的制备
步骤1：向50mL茄形瓶中加入4-(2-溴-4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(int-2a,1g,1.51mmol)，2-(5,6-二氢-4H-吡喃-2-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(0.5g,2.26mmol)，Pd(dppf)Cl2(0.1g,0.15mmol)，磷酸钾(1.0g,4.53mmol)，水(5mL)和二氧六环(20mL)，升高温度到85℃搅拌2小时。浓缩反应液，剩余物快速柱层析(PE:EA＝5:1)得到4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,4-二氢-2H-吡喃-6-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(84-a，641.3mg,0.96mmol,63.8％)。MS Calcd:665.23；MS Found:664.27([M-H]-).
步骤2：向50mL茄形瓶中加入4-(4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(3,4-二氢-2H-吡喃-6-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)哌嗪-1-甲酸叔丁酯(84-a，600mg,0.90mmol)，盐酸甲醇溶液(10mL)，置于50℃下反应2小时。将反应液调至略碱，浓缩，剩余物经快速柱层析(DCM:MeOH＝10:1)得到N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,4-二氢-2H-吡喃-6-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(84-b，260mg,0.46mmol,51.0％)。MS Calcd:565.18；MS Found:566.20([M+H]+).
步骤4：向25mL茄形瓶中加入4-羟基异噁唑-3-甲酸(36.4mg,0.27mmol)，DCM(2mL)，氮气保护下搅拌3小时，再加入DIPEA(0.14mL,0.86mmol)，N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,4-二氢-2H-吡喃-6-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(84-b，50mg,0.09mmol)，室温反应2小时。浓缩反应液，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,4-二氢-2H-吡喃-6-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(84，6mg,0.01mmol,10.0％)。MS Calcd:676.18；MS Found:677.3([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.32(s,1H),8.60(s,1H),8.09(d,J＝8.8Hz,1H),7.97(d,J＝2.0Hz,1H),7.74 –7.71(m,1H),5.89(t,J＝4.0Hz,1H),5.33(s,2H),4.54(d,J＝12.4Hz,1H),4.12(t,J＝5.2Hz,2H),3.64(d,J＝12.4Hz,1H),3.49(t,J＝11.2Hz,2H),3.06-2.99(m,4H),2.84(d,J＝11.2Hz,1H),2.74(d,J＝11.2Hz,1H),2.20(q,J＝5.6Hz,2H),1.87(q,J＝5.6Hz,2H),1.21(t,J＝7.2Hz,3H).
实施例67 (R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物85)的制备
步骤1：将1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,94mg,0.42mmol)和(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(54-e，50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：(R)-N-(5-氯-2-甲基-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(85，13mg,0.02mmol,20％yield).MS Calcd:771.29；MS Found:772.44([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.12(s,1H),7.98(s,1H),7.78(s,1H),7.66(s,1H),6.85–6.83(m,1H),5.30(s,2H),4.71–4.63(m,1H),4.31-4.26(m,3H),3.83(t,J＝5.2Hz,2H),3.71(d,J＝11.2Hz,1H),3.52(t,J＝11.2Hz,1H),3.20–3.16(m,1H),2.99–2.90(m,1H),2.80(d,J＝10.0Hz,1H),2.70–2.66(m,1H),2.54-2.50(m,2H),2.38(s,3H),2.00–1.80(m,8H),1.66-1.62(m,2H),1.45(brs,3H),1.25(t,J＝7.2Hz,3H).
实施例68(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-5-乙基-2-吗啉基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物86)的制备
步骤1：将N-(2-氯-4-(三氟甲基)苯基)-2-碘代乙酰胺(296.5mg,0.82mmol)置于100mL茄形瓶中，加入1,4-二氧六环溶液10mL。再向其中依次加入(R)-4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(int-6,300mg,0.68mmol)和DIPEA(0.34mL,2.04mmol)，75℃下搅拌5小时。直接将反应液浓缩，经柱层析纯化(PE:EA＝1:1)得(R)-4-(2-溴-4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-a，360mg,0.53mmol,78％yield).MS Calcd:677.12；MS Found:678.20([M+H]+).
步骤2：将(R)-4-(2-溴-4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-a，360mg,0.53mmol)置于25mL茄形瓶中，加入DMSO溶液3mL。再向其中依次加入吗啉(0.09mL,1.37mmol)和乙酸钾(134mg,1.37mmol)。加热到120℃搅拌过夜。向反应液中加水(20mL)稀释，EA萃取(30mL×3)，合并有机相，无水硫酸钠干燥，过滤浓缩，粗产品经柱层析纯化(DCM:MeOH＝50：1)得(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-2-吗啉基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-b，180mg，0.26mmol，57％yield)MS Calcd:682.26；MS Found:683.23([M+H]+).
步骤3：将(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-2-吗啉基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-b，180mg，0.26mmol)溶于4mL二氯甲烷溶液中，再向其中加入1mL三氟乙酸，室温下搅拌2小时。将反应液直接浓缩，向残渣中加入20mL饱和碳酸氢钠溶液，用二氯甲烷萃取(20mL×3)，合并有机相，干燥浓缩，得粗品(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(3-甲基哌嗪-1-基)-2-吗啉基-7-氧代-[1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(86-c，150mg,0.26mmol,97％yield)MS Calcd:582.26；MS Found:583.23([M+H]+).
步骤4：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,90mg,0.42mmol)和(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(3-甲基哌嗪-1-基)-2-吗啉基-7-氧代- [1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(86-c，50mg，0.08mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟，直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-5-乙基-2-吗啉基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(86，13mg,0.02mmol,18.6％yield)MS Calcd:746.27；MS Found:747.38([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.35(s,1H),8.07(d,J＝8.4Hz,1H),7.98(d,J＝2.0Hz,1H),7.75(dd,J＝8.8,2.0Hz,1H),7.70(s,1H),5.25(s,2H),4.80(p,J＝8.4Hz,1H),4.62(brs,1H),4.21-4.17(m,1H),3.71-3.68(m,5H),3.51(t,J＝11.6Hz,1H),3.40-3.36(m,5H),3.17-3.09(m,1H),2.96-2.88(m,1H),2.76(d,J＝10.0Hz,1H),2.63(d,J＝11.6Hz,1H),2.58-2.48(m,2H),2.35-2.27(m,2H),1.84-1.76(m,2H),1.47–1.40(m,3H),1.22(t,J＝7.2Hz,3H).
实施例69 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-1-甲基-4-氧代-1,4-二氢吡啶-2-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物87)的制备
步骤1：将3-(苄氧基)-4-氧代-4H-吡喃-2-甲酸(87-a，200mg,0.82mmol)置于25mL茄形瓶中，向其中加入甲胺甲醇溶液(3mL)，80℃加热回流过夜。直接将反应液浓缩，得到粗品3-(苄氧基)-1-甲基-4-氧代-1,4-二氢吡啶-2-甲酸(87-b，200mg，0.78mmol，95％yield).MS Calcd:259.08；MS Found:260.13([M+H]+).
步骤2：将N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,200mg,0.35mmol)，和3-(苄氧基)-1-甲基-4-氧代-1,4-二氢吡啶-2-甲酸(87-b，137mg，0.69mmol)置于25mL茄形瓶中，向其中加入DMF溶液(3mL)，再依次加入HATU(268mg,0.88mmol)，DIPEA(0.18mL,1.33mmol)，室温反应过夜。向反应液中加水(20mL)稀释，EA萃取(30mL×3)，合并有机相，无水硫酸钠干燥，过滤浓缩，粗产品经Prep-TLC纯化(DCM:MeOH＝20：1)得到2-(6-(4-(3-(苄氧基)-1-甲基-4-氧代-1,4-二氢吡啶-2-酰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(87-c，70mg，0.09mmol，25％yield)MS Calcd:806.26；MS Found:807.24([M+H]+).
步骤3：将2-(6-(4-(3-(苄氧基)-1-甲基-4-氧代-1,4-二氢吡啶-2-酰基)哌嗪-1-基)- 2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(87-c，70mg，0.09mmol)置于25mL茄形瓶中，向其中加入DCM溶液(3mL)，再加入对甲苯苯磺酸水合物(82mg，0.43mmol)，50℃反应过夜。将反应液浓缩，经Prep-HPLC纯化，得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(3-羟基-1-甲基-4-氧代-1,4-二氢吡啶-2-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(87，20mg，0.03mmol，30％yield)MS Calcd:716.21；MS Found:717.23([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.37(s,1H),8.08(dd,J＝8.8,2.0Hz,1H),7.97(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),7.63(dd,J＝11.8,7.2Hz,1H),6.86-6.84(m,1H),6.18(dd,J＝7.0,2.0Hz,1H),5.34(s,2H),4.54(t,J＝13.2Hz,1H),4.27(q,J＝2.8Hz,2H),3.72–3.32(m,11H),3.09–2.96(m,3H),2.85(t,J＝10.0Hz,1H),2.76–2.69(m,1H),1.21(t,J＝7.2Hz,3H).
实施例70 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(cis-2,6-二甲基吗啉基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物88)的制备
步骤1：将4-羟基异噁唑-3-甲酸(int-7,32mg,0.25mmol)和1-氯-N,N,2-三甲基丙烯胺(36mg,0.27mmol)分别加入到DCM溶液中(2mL)，室温下搅拌3小时，然后加入N-(2-氯-4-(三氟甲基)苯基)-2-(2-(cis-2,6-二甲基吗啉基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(81-b,50mg,0.08mmol)、DIPEA(65mg,0.5mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(cis-2,6-二甲基吗啉基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(88，7mg,0.01mmol,11.8％yield)。MS Calcd:707.22；MS Found:708.46([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.34(s,1H),8.59(s,1H),8.06(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),5.23(s,2H),4.52(d,J＝12.8Hz,1H),3.89(dd,J＝12.8,2.4Hz,2H),3.66–3.58(m,4H),3.51-3.44(m,2H),3.35–3.30(m,1H),3.04–2.93(m,4H),2.80(d,J＝11.2Hz,1H),2.71–2.68(m,1H),1.19(t,J＝7.2Hz,3H),1.13(d,J＝6.4Hz,6H).
实施例71 (R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三 氟甲基)苯基)乙酰胺(化合物90)的制备
步骤1：称取2-碘-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(234mg,0.68mmol)，(R)-4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(int-6,300mg,0.68mmol)到25mL单口瓶中，加入1,4-二氧六环(5mL)和DIEA(0.34mL,2.04mmol)，加完后80℃反应2小时。减压浓缩，残余物经柱层析分离纯化(DCM：MeOH＝20:1)得(R)-4-(2-溴-5-乙基-4-(2-(2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(90-a,389mg,0.56mmol,82.8％Yield)。MS Calcd:655.17、657.17；MS Found:556.12、558.09([M+H-100]+)。
步骤2：称取(R)-4-(2-溴-5-乙基-4-(2-(2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(90-a,389mg,0.56mmol)，2-(3,6-二氢-2H-吡喃-4-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(177.4mg,0.84mmol)，磷酸钾(388.9mg,1.69mmol)，Pd(dppf)Cl2(82.4mg,0.11mmol)到50mL单口瓶中，加入二氧六环(5mL)和水(2mL)，加完后氮气置换三次，然后90℃反应2小时。将反应液拌样柱层析纯化(DCM：MeOH＝20:1)得到(R)-4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(90-b,312mg,0.43mmol,75.6％Yield)。MS Calcd:659.30；MS Found:604.24([M+H-56]+)。
步骤3：称取(R)-4-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-4-(2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(90-b,312mg,0.43mmol)到50mL单口瓶中，加入DCM(5mL)和三氟乙酸(2mL)，加完后室温搅拌1小时。将反应液浓缩部分溶剂，然后加入饱和碳酸氢钠溶液，乙酸乙酯萃取，有机相除水，浓缩，残余物柱层析纯化 (DCM：MeOH＝10:1)得到(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(90-c,211mg,0.36mmol,84.2％Yield)。MS Calcd:559.25；MS Found:560.20([M+H]+)。
步骤4：称取4-羟基异噁唑-3-甲酸(int-7,16.4mg,0.13mmol)和HATU(102.7mg,0.27mmol)到25mL单口瓶中，加入DCM(1mL)，然后加入DIPEA(0.03mL,0.17mmol)，加完后室温搅拌5分钟后加入(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(90-c,50mg,0.08mmol)，加完后室温反应2小时。将反应液加入2mL氯化铵水溶液，分离有机相，浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基异噁唑-3-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(90,25mg,0.04mmol,43.1％Yield)。MS Calcd:670.25；MS Found:671.33([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.02(s,1H),9.78(s,1H),8.60(d,J＝1.6Hz,1H),7.74(d,J＝8.4Hz,1H),7.64(d,J＝2.0Hz,1H),7.55(d,J＝8.4Hz,1H),6.86-6.84(m,1H),5.27(s,2H),4.28(d,J＝2.9Hz,2H),3.88-3.81(m,3H),3.69(d,J＝11.6Hz,1H),3.54–3.43(m,2H),3.32–3.17(m,3H),2.96-2.90(m,1H),2.84(d,J＝11.2Hz,1H),2.73–2.63(m,2H),2.37(s,3H),1.43(dd,J＝6.8,2.0Hz,3H),1.24(t,J＝7.2Hz,3H).
实施例72 2-(6-((R)-4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物91)的制备
步骤1：将(R)-4-(2-溴-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(int-6,0.3g,0.68mmol)、N-(5-氟-2-甲基-4-(三氟甲基)苯基)-2-碘乙酰胺(294mg,0.82mmol)和DIPEA(264mg,2.04mmol)分别加入到二氧六环溶液中(10mL)，于80℃条件下搅拌4小时。将反应液减压浓缩，残 余物经硅胶柱层析(PE:EA＝1:1)分离纯化得(R)-4-(2-溴-5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(91-a,0.2g,0.3mmol,43.6％yield)。MS Calcd:673.16；MS Found:574.20([M-100+H]+).
步骤2：将(R)-4-(2-溴-5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(91-a,0.2g,0.3mmol)、2-(4-甲氧基环己-1-烯-1-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(307mg,1.29mmol)、Pd(dppf)Cl2·CH2Cl2(70mg,0.09mmol)和磷酸钾(274mg,1.29mmol)依次加入10mL二氧六环/水(4:1)混合溶液中，抽真空氮气置换3次，升温至80℃搅拌2h。将反应液完全浓缩，残余物经硅胶柱层析(PE:EA＝1:1)分离纯化得(2R)-4-(5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧乙基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(91-b,190mg,0.27mmol,62.6％yield，非对映体混合物)。MS Calcd:705.33；MS Found:650.25([M-56+H]+).
步骤3：将(2R)-4-(5-乙基-4-(2-((5-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧乙基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(91-b,190mg,0.27mmol)加入盐酸-二氧六环溶液中(4mL)，置于室温搅拌过夜。将反应液完全浓缩，向残余物中加入饱和碳酸氢钠溶液，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得粗品2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-6-((R)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(91-c,0.12g,0.2mmol,73.6％yield，非对映体混合物)。MS Calcd:605.27；MS Found:606.31([M+H]+).
步骤4：将1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,87mg,0.41mmol)、DIPEA(32mg,0.25mmol)和2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-6-((R)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(91-c,50mg,0.08mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，残余物经Prep-HPLC纯化得到标题化合物：2-(6-((R)-4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(91,18mg,0.02mmol,26.7％yield，非对映体混合物)。MS Calcd:769.33；MS Found:770.34([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.12(s,1H),8.24(s,1H),7.80(d,J＝12.8Hz,1H),7.68(d,J＝8.0Hz,1H),7.36(s,1H),6.74-6.72(m,1H),5.30(s,2H),4.83–4.60(m,2H),4.24–4.15(m,1H),3.68-3.15(m,8H),2.97–2.93(m,1H),2.75–2.68(m,1H),2.60–2.34(m,6H), 2.19-2.12(m,4H),2.00–1.94(m,2H),1.74-1.66(m,4H),1.47-1.43(m,3H),1.23(t,J＝7.2Hz,3H).
实施例73 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,4-二氢-2H-吡唑-6-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物92)的制备
步骤1：向25mL茄形瓶中加入N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,4-二氢-2H-吡喃-6-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(84-b，60mg,0.11mmol)，1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,22.3mg,0.11mmol)，2-甲基-2-丁醇(2mL)和DIPEA(0.04mL)，氮气置换3次，180℃搅拌反应2小时，向其中加入1mL甲醇溶解，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)哌嗪-1-基)-2-(3,4-二氢-2H-吡唑-6-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(92,18.2mg,0.02mmol,22.6％).MS Calcd:729.24；MS Found:730.30([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.36(s,1H),8.10(d,J＝8.4Hz,1H),7.99(d,J＝2.0Hz,1H),7.76–7.70(m,2H),5.89(t,J＝4.0Hz,1H),5.34(s,2H),4.80(p,J＝8.4Hz,1H),4.34(d,J＝12.6Hz,2H),4.12(dd,J＝6.0,4.0Hz,2H),3.52-3.45(m,2H),3.21-3.14(m,2H),3.02(q,J＝7.2Hz,2H),2.78(d,J＝11.2Hz,2H),2.52(p,J＝1.9Hz,2H),2.35–2.27(m,2H),2.22-2.18(m,2H),1.89-1.77(m,4H),1.22(t,J＝7.2Hz,3H),羟基未出峰.
实施例74 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-吗啉基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物93)的制备
步骤1：将1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,115mg,0.51mmol)和(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(3-甲基哌嗪-1-基)-2-吗啉基-7-氧代- [1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(86-c，60mg，0.1mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，残余物经Prep-HPLC纯化得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-吗啉基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(93，25mg,0.03mmol,32％yield).MS Calcd:760.28；MS Found:761.34([M+H]+).1H NMR(400MHz,DMSO-d6)δ10.35(s,1H),8.06(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.67(s,1H),5.24(s,2H),4.70–4.63(m,1H),3.72-3.66(m,6H),3.50(t,J＝6.4Hz,1H),3.41-3.38(m,6H),3.16-3.07(m,1H),2.96-2.86(m,1H),2.76(d,J＝10.0Hz,1H),2.63(d,J＝11.6Hz,1H),2.03–1.74(m,6H),1.67-1.57(m,2H),1.43(d,J＝6.4Hz,3H),1.22(d,J＝7.2Hz,3H)，羟基未出峰.
实施例75 (R)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物94)的制备
步骤1：称取1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(17-b,85mg,0.40mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(90-c,50mg,0.08mmol)到25mL单口瓶中，加入2-甲基-2-丁醇(2mL)，然后加入DIPEA(0.04mL,0.24mmol)，加完后升温到180℃反应2小时。将反应液浓缩，残余物制备纯化得到标题化合物：(R)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(94,4.0mg,0.01mmol,6.8％Yield)。MS Calcd:723.31；MS Found:724.30([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.03(s,1H),7.74(d,J＝8.4Hz,1H),7.67-7.64(m,2H),7.57–7.54(m,1H),6.86–6.84(m,1H),5.28(s,2H),4.84-4.76(m,1H),4.30-4.24(m,3H),3.83(t,J＝5.6Hz,2H),3.70(d,J＝11.6Hz,1H),3.52-3.48(m,2H),3.21-3.16(m,2H),3.00-2.95(m,1H),2.79(d,J＝10.0Hz,1H),2.70–2.65(m,1H),2.56-2.48(m,2H),2.38(s,3H),2.32-2.26(m,2H),1.83-1.75(m,2H),1.45(brs,3H),1.31-1.24(m,5H).
实施例76 (R)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2- (3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物95)的制备
步骤1：称取1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,90mg,0.40mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(90-c,50mg,0.08mmol)到25mL单口瓶中，加入2-甲基-2-丁醇(2mL)，然后加入DIPEA(0.04mL,0.24mmol)，加完后升温到180℃反应2小时。将反应液浓缩，残余物制备纯化得到标题化合物：(R)-2-(6-(4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡嗪-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(95,18mg,0.02mmol,29.7％Yield)。MS Calcd:737.33；MS Found:738.38([M+H]+)。1H NMR(400MHz,DMSO-d6，重水交换)δ7.72(d,J＝8.4Hz,1H),7.66(s,1H),7.63(d,J＝2.0Hz,1H),7.55(d,J＝8.4Hz,1H),6.86–6.84(m,1H),5.26(s,2H),4.69-4.62(m,1H),4.27(q,J＝2.8Hz,2H),3.82(t,J＝5.6Hz,2H),3.69-3.49(m,2H),3.19-3.10(m,1H),2.98-2.92(m,1H),2.80(d,J＝10.0Hz,1H),2.69-2.66(m,1H),2.36(s,3H),2.54-2.50(m,1H),2.03-1.99(m,2H),1.88–1.78(m,4H),1.65–1.61(m,2H),1.43(brs,3H),1.27–1.22(m,5H).
实施例77 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-苯基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物96)的制备
步骤1：将乙氧基甲叉丙二酸二乙酯(3-a,1.49g,6.92mmol)、苯肼盐酸盐(1.0g，6.92mmol)和碳酸钾(2.86g,20.75mmol)分别加入到水溶液中(10mL)，置于100℃条件下搅拌6小时。用5N HCl调至pH＝2，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得5-羟基-1-苯基-1H-吡唑-4-甲酸乙酯(96-a,0.63g,2.71mmol,39.2％yield)。MS Calcd:232.08；MS Found:233.04([M+H]+).
步骤2：将5-羟基-1-苯基-1H-吡唑-4-甲酸乙酯(96-a,100mg,0.43mmol)、DIPEA(34mg,0.27mmol)和(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,50mg,0.09mmol)加入2-甲基-2-丁醇溶液中(2mL)，油浴升温至180℃搅拌30min。将反应液完全浓缩，剩余物经Prep-HPLC纯化，得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(5-羟基-1-苯基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(96,16mg,0.02mmol,23.5％yield)。MS Calcd:765.24；MS Found:766.24([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.02(d,J＝8.4Hz,1H),7.94(d,J＝2.0Hz,1H),7.90-7.85(m,1H),7.72-7.69(m,2H),7.44-7.40(m,2H),7.23-7.20(m,1H),6.83(s,1H),5.31(s,2H),4.67(brs,1H),4.24(brs,3H),3.84-3.80(m,4H),3.68(d,J＝10.8Hz,1H),3.48(d,J＝12.0Hz,1H),3.18-3.12(m,1H),2.95-2.90(m,1H),2.78(d,J＝9.2Hz,1H),2.65(d,J＝11.2Hz,1H),2.54-2.51(m,2H),1.44(brs,3H),1.23(t,J＝7.2Hz,3H).
实施例78 2-(6-((R)-4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物98)的制备
步骤1：将2-(2-溴-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(56-a,1g,2.1mmol)，2-(4-甲氧基环己-1-烯-1-基)-4,4,5,5-四甲基-1,3,2-二氧杂硼烷(int-8,760mg,3.2mmol)，磷酸钾(1.4g,6.3mmol)，1,1-双(二苯基膦)二荗铁二氯化钯(150mg,0.21mmol)加入到二氧六环(15mL)和水(4mL)的混合溶剂中，氮气置换3分钟，升高温度到85℃，搅拌3小时，减压浓缩，剩余物倒入水中，用乙酸乙酯萃取(20mLx3)，合并有机层，经减压浓缩，剩余物经柱层析纯化(EA：PE＝1:2)得到2-(5-乙基-2-(4-甲氧基环己-1-烯 -1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98-a,730mg,1.43mmol，68％)MS Calcd:507.19；MS Found:508.21([M+H]+)。
步骤2：将2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98-a,730mg,1.43mmol)加入到DMF中(10ml)，然后加入NBS(255mg,1.43mmol)，升高温度到60℃，搅拌3小时。然后将反应体系倒入水中，用EA萃取(20mLx3)，有机层经无水硫酸钠干燥，抽滤，浓缩，剩余物经柱层析纯化(EA：PE＝1:1)得到2-(6-溴-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98-b,510mg,0.87mmol，61％)MS Calcd:585.10；MS Found:586.15([M+H]+)。
步骤3：将2-(6-溴-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98-b,510mg,0.87mmol)，(R)-2-甲基哌嗪-1-甲酸叔丁酯(1.7g，8.7mmol)，四氟硼酸银(170mg，0.87mmol)加入到干燥的DMSO(5mL)中，然后氮气保护下，升高温度到120℃，搅拌4小时，原料反应完毕，冷却到室温，加入饱和碳酸氢钠水溶液，然后用乙酸乙酯萃取(10mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，剩余物经柱层析纯化(EA：PE＝1:1)，得到(2R)-4-(5-乙基-4-(2-((3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧乙基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(98-c,410mg,0.58mmol，66％,非对映体混合物)。MS Calcd:705.33；MS Found:706.38([M+H]+)。
步骤4：将(2R)-4-(5-乙基-4-(2-((3-氟-2-甲基-4-(三氟甲基)苯基)氨基)-2-氧乙基)-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(98-c,410mg,0.58mmol)加入到DCM中(10mL)，然后加入三氟乙酸(3mL)，在室温下搅拌4小时，原料反应完毕，减压浓缩，剩余物加入饱和碳酸氢钠水溶液，然后用DCM萃取(10mLx3)，有机层经无水硫酸钠干燥，抽滤，滤液经减压浓缩，得2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-6-((R)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98-d,230mg,0.38mmol,65％yield,对映体混合物)。MS Calcd:605.27；MS Found:606.31([M+H]+)。
步骤5：将2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-6-((R)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98-d,60mg,0.1mmol)，DIPEA(15mg，0.10mmol)，1-环丁基-5-羟基-1H-吡唑-4-甲酸乙酯(105mg，0.5mmol)加入到2-甲基-2-丁醇(1mL)中，油浴升温到180℃，搅拌30分钟，将反应体系溶解到甲醇中，经prep-HPLC纯化，得到标题化合物： 2-(6-((R)-4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98,15mg,0.019mmol,19.0％yield,非对映体混合物)。MS Calcd:769.33；MS Found:770.38([M+H]+)。1H NMR(400MHz,DMSO-d6，重水交换)δ7.68(s,1H),7.63–7.56(m,2H),6.74(brs,1H),5.26(s,2H),4.81-4.76(m,1H),3.68-3.45(m,5H),3.29(s,3H),3.18-3.11(m,1H),2.96-2.91(m,1H),2.78(d,J＝10.0Hz,1H),2.66(d,J＝11.6Hz,1H),2.58-2.40(m,4H),2.35–2.13(m,7H),1.96-1.92(m,1H),1.82-1.65(m,2H),1.42(brs,3H),1.24-1.21(m,5H).
实施例79 2-(6-((R)-4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物99)的制备
步骤1：将2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-6-((R)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(98-d,60mg,0.1mmol)，DIEA(15mg，0.10mmol)，1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,110mg，0.5mmol)加入到2-甲基-2-丁醇(1mL)中，油浴升温到180℃，搅拌30分钟，将反应体系溶解到甲醇中，经prep-HPLC纯化，得到标题化合物：2-(6-((R)-4-(1-环戊基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(99,11mg,0.014mmol,14.0％yield，非对映体混合物)。MS Calcd:783.35；MS Found:784.39([M+H]+)。1H NMR(400MHz,DMSO-d6)δ10.23(s,1H),7.65–7.58(m,3H),6.74(d,J＝4.0Hz,1H),5.27(s,2H),4.68–4.63(m,1H),3.70(d,J＝11.6Hz,1H),3.56-3.47(m,2H),3.31-3.12(m,4H),2.99-2.93(m,1H),2.78(d,J＝10.0Hz,1H),2.69–2.64(m,1H),2.55-2.41(m,2H),2.26–2.14(m,5H),2.02–1.79(m,9H),1.72–1.61(m,4H),1.43(brs,3H),1.26-1.22(t,J＝7.2Hz,3H).
实施例80 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-2-(4-(二甲基氨基)哌啶-1-基)-5-乙基-7-氧代-[1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物101)的制备
步骤1：向100mL茄形瓶中加入(R)-4-(2-溴-4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-a，300mg,0.44mmol)，4-(二甲基氨基)六氢吡啶(284mg,2.22mmol)和溶剂DMSO(5mL)。然后向其中加乙酸钾(217mg，2.22mmol)，120℃搅拌过夜。加入30mL水稀释，用DCM萃取(30mL×3)，饱和食盐水洗涤有机相3次，无水硫酸钠干燥，减压旋干得到粗品，粗品通过柱层析纯化(DCM:MeOH＝10:1)，得(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(4-(二甲基氨基)哌啶-1-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(101-a,200mg,0.28mmol,62.3％yield).MS Calcd:723.32.；MS Found:724.37([M+H]+).
步骤2：向100mL茄形瓶中加(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-2-(4-(二甲基氨基)哌啶-1-基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(101-a,200mg,0.28mmol)溶剂DCM(2mL)。然后向其中加TFA(0.5mL)，室温搅拌2小时。加入饱和碳酸氢钠溶液将pH调至碱性，用DCM萃取(20mL×3)，饱和食盐水洗涤有机相，无水硫酸钠干燥，减压旋干得粗品(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(4-(二甲基氨基)哌啶-1-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(101-b,120mg,0.19mmol,69.6％yield)。MS Calcd:623.27；MS Found:624.31([M+H]+).
步骤3：向10mL微波管中加入(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(4-(二甲基氨基)哌啶-1-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(101-b,50mg,0.08mmol)和溶剂2-甲基-2-丁醇(1mL)。然后向其中加1-环丁基-5-羟基吡唑-4-甲酸乙酯(17-b,67mg,0.32mmol)和DIPEA(31mg,0.24mmol)，180℃反应1小时。将溶剂旋干，向其中加入甲醇(1mL)，粗品通过Prep-HPLC纯化得到标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-2-(4-(二甲基氨基)哌啶-1-基)-5-乙基-7-氧代-[1，2，4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(101,10mg,0.01mmol, 15.4％yield)。MS Calcd:787.33；MS Found:788.20([M+H]+)。1H NMR(400MHz,DMSO-d6，重水交换)δ8.01(d,J＝8.4Hz,1H),7.94(s,1H),7.72(d,J＝8.4Hz,1H),7.42(s,1H),5.20(s,2H),4.75(p,J＝8.4Hz,1H),4.61(brs,1H),4.16(d,J＝12.8Hz,2H),3.63(d,J＝10.4Hz,1H),3.44(d,J＝11.6Hz,1H),3.17–3.03(m,3H),2.92-2.86(m,3H),2.70-2.62(m,7H),2.43–2.34(m,3H),2.16(d,J＝9.2Hz,2H),1.98(d,J＝11.6Hz,2H),1.72–1.64(m,3H),1.55-1.46(m,2H),1.35(d,J＝6.0Hz,3H),1.19(t,J＝7.2Hz,3H).
实施例81 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-7-氧代-2-(吡啶-3-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物103)的制备
步骤1：向50mL茄形瓶中加入(R)-4-(2-溴-4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-a，500mg,0.74mmol)，吡啶-3-基硼酸(136.2mg,1.11mmol)，Pd(dppf)Cl2(53.6mg,0.07mmol)，磷酸钾(509.7mg,2.22mmol)，水(5mL)和二氧六环(20mL)，升高温度到85℃，搅拌3小时。浓缩反应液，剩余物快速柱层析(PE:EA＝4:1)得(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-2-(吡啶-3-基)-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(103-a，403.5mg,0.60mmol,80.9％)。MS Calcd:674.23；MS Found:675.26([M+H]+).
步骤2：向50mL茄形瓶中加入(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-2-(吡啶-3-基)-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(103-a，400mg,0.59mmol)和盐酸甲醇溶液(10ml)，室温搅拌2小时。向反应液中加入饱和碳酸氢钠，调至pH＝7，浓缩反应液，加水(50mL)溶解，DCM(3*30mL)萃取，浓缩得粗品(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-2-(吡啶-3-基)-[1,2,4]三唑并[1,5-a]嘧啶- 4(7H)-基)乙酰胺(103-b，321.3mg,0.56mmol,94.3％)。MS Calcd:574.18；MS Found:575.17([M+H]+).
步骤3：向25mL茄形瓶中加入(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-2-(吡啶-3-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(103-b，50mg,0.09mmol)，1-环丁基-5-羟基吡唑-4-甲酸乙酯(36.6mg,0.17mmol)，2-甲基-2-丁醇(2mL)溶液中和DIPEA(0.04mL)抽真空氮气置换3次，180℃反应2小时。浓缩反应液，加2mL甲醇溶解，经prep-HPLC纯化，得标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(6-(4-(1-环丁基-5-羟基-1H-吡唑-4-酰基)-3-甲基哌嗪-1-基)-5-乙基-7-氧代-2-(吡啶-3-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(103，9mg,0.01mmol,11.7％yield)。MS Calcd:738.24；MS Found:739.0([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ9.24(d,J＝2.0Hz,1H),8.70(dd,J＝4.8,1.6Hz,1H),8.45-8.42(m,1H),8.03(d,J＝8.4Hz,1H),7.96(d,J＝2.0Hz,1H),7.73–7.69(m,2H),7.59(dd,J＝8.0,4.8Hz,1H),5.40(s,2H),4.78(p,J＝8.4Hz,1H),3.72-3.67(m,1H),3.53-3.47(m,1H),3.21-3.16(m,1H),3.00-2.93(m,1H),2.82(d,J＝9.6Hz,1H),2.71–2.68(m,1H),2.58-2.44(m,3H),2.32–2.28(m,2H),1.82-1.74(m,2H),1.43(brs,3H),1.27-1.21(m,5H).
实施例82 N-(2-氯-4-(三氟甲基)苯基)-2-(6-(((R)-4-(1-环戊基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物104)的制备
步骤1：将化合物(R)-4-(2-溴-4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-a，300mg,0.44mmol)和2-(4-甲氧基环己-1-烯-1-基)-4,4,5-5-四甲基-1,3,2-二氧杂硼烷(211m g,0.89mmol)溶于1,4-二氧六环(10mL)和H2O(2mL)中，再依次加入Pd(dppf)Cl2(32mg,0.04mmol)，磷酸钾(305mg,1.33mmol)，置换三次氮气，80℃反应5.0小时。将反应液浓缩，向残渣中加入30mL乙酸乙酯，经硅藻土过滤，滤液浓缩，经柱层析纯化(DCM:MeOH＝50：1)得(2R)-4-(4-(2-((2-氯-4- (三氟甲基)苯基)氨基)-2-氧乙基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(104-a，130mg，0.18mmol，41％yield，非对映体混合物).MS Calcd:707.28；MS Found:708.30([M+H]+).
步骤2：将(2R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧乙基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(104-a，130mg，0.18mmol)溶于4mL二氯甲烷溶液中，再向其中加入1mL三氟乙酸，室温下搅拌2小时。将反应液直接浓缩，向残渣中加入20mL饱和碳酸氢钠溶液，用二氯甲烷萃取(20mL×3)，合并有机相，干燥浓缩，得粗品：N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-6-((R)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(104-b，105mg，0.17mmol，94％yield，非对映体混合物)MS Calcd:607.23；MS Found:608.20([M+H]+).
步骤3：将N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基环己-1-烯-1-基)-6-((R)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(104-b，50mg,0.08mmol)和1-环戊基-5-羟基-1H-吡唑-4-甲酸乙酯(41-a,92mg,0.41mmol)置于25mL茄形瓶中，加入2mL 2-甲基-2-丁醇。随后在180℃下，搅拌30分钟。直接将反应液浓缩，经Prep-HPLC纯化得到标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(6-(((R)-4-(1-环戊基-5-羟基-1H-吡唑-4-羰基)-3-甲基哌嗪-1-基)-5-乙基-2-(4-甲氧基环己-1-烯-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺
(104，35mg，0.04mmol，52％yield，非对映体混合物)。MS Calcd:785.30；MS Found:786.33([M+H]+).1H NMR(400MHz,DMSO-d6,重水交换)δ8.08(d,J＝8.8Hz,1H),7.98(d,J＝2.0Hz,1H),7.74(dd,J＝8.8,2.0Hz,1H),7.60(s,1H),6.75(t,J＝4.0Hz,1H),5.33(s,2H),4.65(q,J＝7.2Hz,1H),3.69(d,J＝11.2Hz,1H),3.54-3.51(m,2H),3.30(s,3H),3.18-3.12(m,1H),2.98-2.92(m,1H),2.79(d,J＝9.6Hz,1H),2.66(d,J＝11.2Hz,1H),2.56-2.42(m,3H),2.17(d,J＝18.8Hz,1H),1.98–1.78(m,9H),1.71–1.62(m,4H),1.44(brs,3H),1.24(t,J＝7.2Hz,3H).
实施例83 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(4-羟基异恶唑-3-羰基)-3-甲基哌嗪-1-基)-2-(4-甲氧基哌啶-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物106)的制备
步骤1：将(R)-4-(2-溴-4-(2-(2-氯-4-(三氟甲基)苯基)氨基)-2-氧代乙基)-5-乙基-7-氧代-4,7-二氢-[1,2,4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(86-a，0.6g,0.89mmol)、4-甲氧基哌啶(0.51g，4.43mmol)和乙酸钾(0.52g,5.32mmol)分别加入到DMSO溶液中(3mL)，升温至120℃条件下搅拌过夜。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:1)分离纯化得(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧乙基)-5-乙基-2-(4-甲氧基哌啶-1-基)-7-氧代-4,7-二氢-[1，2，4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(106-a,0.4g,0.56mmol,63.5％yield)。MS Calcd:710.29；MS Found:655.22([M-56+H]+)。
步骤2：将(R)-4-(4-(2-((2-氯-4-(三氟甲基)苯基)氨基)-2-氧乙基)-5-乙基-2-(4-甲氧基哌啶-1-基)-7-氧代-4,7-二氢-[1，2，4]三唑并[1,5-a]嘧啶-6-基)-2-甲基哌嗪-1-甲酸叔丁酯(106-a,0.4g,0.56mmol)加入盐酸-二氧六环溶液中(4mL)，置于室温搅拌过夜。将反应液完全浓缩，向残余物中加入饱和碳酸氢钠溶液，二氯甲烷萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得产物(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基哌啶-1-基)-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(106-b,0.2g,0.33mmol,58.2％yield)。MS Calcd:610.24；MS Found:611.24([M+H]+).
步骤3：将4-羟基异噁唑-3-甲酸(int-7,51mg,0.39mmol)和1-氯-N,N,2-三甲基丙烯胺(55mg,0.41mmol)分别加入到DCM溶液中(2mL)，室温下搅拌3小时，然后加入(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-2-(4-甲氧基哌啶-1-基)-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(106-b,60mg,0.10mmol)、DIPEA(76mg,0.59mmol)，继续升温至40℃搅拌5h。向反应液中加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经高压制备纯化得标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(5-乙基-6-(4-(4- 羟基异恶唑-3-羰基)-3-甲基哌嗪-1-基)-2-(4-甲氧基哌啶-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(106，8mg,0.01mmol,11.2％yield)。MS Calcd:721.24；MS Found:722.32([M+H]+).1H NMR(400MHz,DMSO-d6,重水交换)δ8.40(s,1H),8.02(d,J＝8.8Hz,1H),7.95(d,J＝2.0Hz,1H),7.72(dd,J＝8.8,2.0Hz,1H),5.20(s,2H),3.79-3.73(m,2H),3.67–3.63(m,1H),3.51–3.36(m,3H),3.26(s,3H),3.21-3.06(m,4H),2.88–2.78(m,2H),2.69–2.65(m,1H),1.88–1.83(m,2H),1.44-1.35(m,6H),1.18(t,J＝7.2Hz,3H).
实施例84 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(2-羟基吡唑并[1,5-a]吡啶-3-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1.5-a]嘧啶-4(7H)-基)乙酰胺(化合物107)的制备
步骤1：将1-氨基吡啶碘(107-a,6000mg,27.02mmol)、丙二酸二乙酯(6492.5mg,40.54mmol)和K2CO3(9336.6mg,67.56mmol)依次加入EtOH(100mL)溶液中，置于室温搅拌过夜。将反应过滤，滤液减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得1-(3-乙氧基-3-氧代丙酰氨基)吡啶-1-鎓(107-b,3200mg,9.52mmol,56.6％)。MS Calcd:209.09([M-I]+)；MS Found:209.11([M-I]+).
步骤2：将1-(3-乙氧基-3-氧代丙酰氨基)吡啶-1-鎓(107-b,2800mg,8.33mmol)溶于THF(20mL)溶液中，室温分批加入t-BuOK(1854.4mg,16.53mmol)，置于室温搅拌30min。将反应液完全浓缩，向残余物中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-羟基吡唑并[1,5-a]吡啶-3-甲酸乙酯(107-c,1500mg,7.28mmol,52.9％)。MS Calcd:206.07；MS Found:207.06([M+H]+).
步骤3：将2-羟基吡唑并[1,5-a]吡啶-3-甲酸乙酯(107-c,500mg,2.43mmol)和NaOH(486.0mg,12.15mmol)依次加入EtOH/H2O(6mL，2:1)混合溶液中，升温至80℃搅拌过夜。将反应液完全浓缩，向残余物中缓慢加入5N HCl溶液，直至固体析出，抽滤得2-羟基吡唑并[1,5-a]吡啶-3-甲酸(107-d,200mg,1.12mmol,46.3％)。MS Calcd:178.04；MS Found:179.08([M+H]+).
步骤4：将2-羟基吡唑并[1,5-a]吡啶-3-甲酸(107-d,50mg,0.28mmol)、HATU(160mg,0.42mmol)、DIPEA(90mg,0.7mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,190mg,0.34mmol)加入DCM溶液中(6mL)，室温搅拌过夜。将反应液完全浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(2-羟基吡唑并[1,5-a]吡啶-3-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1.5-a]嘧啶-4(7H)-基)乙酰胺(107,25mg,0.03mmol,11.9％yield)。MS Calcd:725.21；MS Found:726.23([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.46(d,J＝6.8Hz,1H),8.06(d,J＝8.4Hz,1H),7.96(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),7.60(d,J＝8.8Hz,1H),7.37-7.32(m,1H),6.90–6.83(m,2H),5.32(s,2H),4.26(d,J＝2.8Hz,2H),4.19(d,J＝12.4Hz,2H),3.81(t,J＝5.6Hz,2H),3.54-3.47(m,2H),3.17-3.10(m,2H),3.01(q,J＝7.2Hz,2H),2.75(d,J＝10.8Hz,2H),2.53-2.51(m,2H),1.22(t,J＝7.2Hz,3H).
实施例85 2-(6-(4-(7-氯-2-羟基吡唑并[1,5-a]吡啶-3-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1.5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(化合物108)的制备
步骤1：将2-羟基吡唑并[1,5-a]吡啶-3-甲酸乙酯(107-c,1.0g,4.85mmol)、Boc酸酐(1.16g,5.33mmol)和碳酸铯(3.55g,10.91mmol)依次加入10mL DMF溶液中，升温至80℃搅拌8h。向残余物中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(PE:EA＝4:1)分离纯化得2-((叔丁氧羰基)氧基)吡唑并[1,5-a]吡啶-3-甲酸乙酯(108-a,640mg,2.09mmol,43.1％yield)。MS Calcd:306.12；MS Found:207.1([M-100+H]+).
步骤2：将2-((叔丁氧羰基)氧基)吡唑并[1,5-a]吡啶-3-甲酸乙酯(108-a,480mg,1.57mmol)溶于4mL THF溶液中，置于-78℃条件下，缓慢滴加LiHMDS(2.51mL,2.51mmol)，滴毕，在此温度下继续搅拌1h。然后加入六氯乙烷(445mg,1.88mmol)的THF(4mL)溶液，继续搅拌30min，缓慢升至室温搅拌1h。向 反应液中加入饱和氯化铵溶液终止反应，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(PE:EA＝4:1)分离纯化得2-((叔丁氧羰基)氧基)-7-氯吡唑并[1,5-a]吡啶-3-甲酸乙酯(108-b,400mg,1.17mmol,74.9％yield)。MS Calcd:340.08；MS Found:241.1([M-100+H]+).
步骤3：将2-((叔丁氧羰基)氧基)-7-氯吡唑并[1,5-a]吡啶-3-甲酸乙酯(108-b,350mg,1.03mmol)加入2mL二氯甲烷中，然后加入1mL三氟乙酸，置于室温搅拌2h。将反应液完全浓缩，向残余物中加入饱和碳酸氢钠溶液，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(100％EA)分离纯化得7-氯-2-羟基吡唑并[1,5-a]吡啶-3-甲酸乙酯(108-c,220mg,0.91mmol,89.0％yield)。MS Calcd:240.03；MS Found:240.99([M+H]+).
步骤4：将7-氯-2-羟基吡唑并[1,5-a]吡啶-3-甲酸乙酯(108-c,100mg,0.42mmol和氢氧化钠(83mg,2.08mmol)依次加入2mL EtOH/H2O(5:1)混合溶液中，升温至80℃搅拌4h。TLC监测反应完成，将反应液完全浓缩得7-氯-2-羟基吡唑并[1,5-a]吡啶-3-甲酸(108-d,83mg,0.29mmol,70.0％yield)。MS Calcd:212.00；MS Found:213.04([M+H]+).
步骤5：将7-氯-2-羟基吡唑并[1,5-a]吡啶-3-甲酸(108-d,55mg,0.26mmol)、HATU(118mg,0.31mmol)、DIPEA(83mg,0.65mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,160mg,0.28mmol)加入DMF溶液中(2mL)，室温搅拌过夜。向反应液中加入乙酸乙酯萃取，减压浓缩，残余物经prep-HPLC纯化得标题化合物：2-(6-(4-(7-氯-2-羟基吡唑并[1,5-a]吡啶-3-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1.5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(108,16mg,0.02mmol,7.8％yield)。MS Calcd:759.17；MS Found:760.21([M+H]+).1H NMR(400MHz,DMSO，重水交换)δ7.96(d,J＝8.4Hz,1H),7.86(s,1H),7.63(d,J＝8.8Hz,1H),7.52(d,J＝8.8Hz,1H),7.27(t,J＝8.4Hz,1H),7.06(d,J＝7.2Hz,1H),6.75(s,1H),5.23(s,2H),4.18–4.09(m,4H),3.72(t,J＝5.6Hz,2H),3.67(s,2H),3.44-3.37(m,2H),3.06(t,J＝12.0Hz,2H),3.01(q,J＝7.2Hz,2H),2.75(d,J＝10.8Hz,2H),1.21(t,J＝7.2Hz,3H).
实施例86 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(2-羟基咪唑并[1,2-a]吡啶-3-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物109)的制备
步骤1：将2-氨基吡啶(109-a,2.0g,21.25mmol)和溴代丙二酸二乙酯(6.3g,26.57mmol)依次加入40mL乙醇溶液中，升温至回流条件下搅拌过夜。将反应液完全浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-羟基咪唑并[1,2-a]吡啶-3-甲酸乙酯(109-b,3.2g,15.52mmol,73.0％yield)。MS Calcd:206.07；MS Found:207.10([M+H]+).
步骤2：将2-羟基咪唑并[1,2-a]吡啶-3-甲酸乙酯(109-b,2.1g,10.18mmol)、1-(氯甲基)-4-甲氧基苯(1.9g,12.22mmol)和碳酸铯(8.29g,25.46mmol)依次加入40mL DMF溶液中，升温至60℃搅拌5h。向反应液中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(PE:EA＝5:1)分离纯化得2-((4-甲氧基苄基)氧基)咪唑并[1,2-a]吡啶-3-甲酸乙酯(109-c,0.5g,1.53mmol,15.0％yield)。MS Calcd:326.13；MS Found:327.10([M+H]+).
步骤3：将2-((4-甲氧基苄基)氧基)咪唑并[1,2-a]吡啶-3-甲酸乙酯(109-c,90mg,0.28mmol)和氢氧化钠(55mg,1.38mmol)依次加入2mL EtOH/H2O(5:1)混合溶液中，升温至80℃搅拌4h。将反应液完全浓缩得2-((4-甲氧基苄基)氧基)咪唑并[1,2-a]吡啶-3-甲酸(109-d,83mg,0.28mmol,99.4％yield)。MS Calcd:298.10；MS Found:299.1([M+H]+).
步骤4：将2-((4-甲氧基苄基)氧基)咪唑并[1,2-a]吡啶-3-甲酸(109-d,83mg,0.28mmol)、PyBOP(237mg,0.46mmol)、DIPEA(98mg,0.76mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2，180mg,0.3mmol)加入DMF溶液中(4mL)，室温搅拌过夜。向反应液中加入水有固体析出，抽滤得N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(2-((4-甲氧基苄基)氧基)咪唑并[1,2-a]吡啶-3-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(109-e,130mg,0.11mmol,51.3％yield)。MS Calcd:845.27；MS Found:846.3([M+H]+).
步骤5：将N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基- 6-(4-(2-((4-甲氧基苄基)氧基)咪唑并[1,2-a]吡啶-3-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(109-e,130mg,0.11mmol)溶于2mL三氟乙酸中，然后加入苯甲硫醚(80mg,0.65mmol)置于室温搅拌4h。将反应液完全浓缩，加入饱和碳酸氢钠溶液，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(2-羟基咪唑并[1,2-a]吡啶-3-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(109,8mg,0.01mmol,9.7％yield)。MS Calcd:725.21；MS Found:726.21([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ9.01(d,J＝6.8Hz,1H),8.07(d,J＝8.4Hz,1H),7.97(d,J＝2.0Hz,1H),7.73(dd,J＝8.8,2.0Hz,1H),7.53-7.49(m,1H),7.29(d,J＝8.4Hz,1H),7.17-7.13(m,1H),6.85-6.83(m,1H),5.33(s,2H),4.28-4.23(m,4H),3.81(t,J＝5.6Hz,2H),3.57-3.53(m,2H),3.12–2.99(m,4H),2.73(d,J＝10.8Hz,2H),2.53-2.51(m,2H),1.23(t,J＝7.2Hz,3H).
实施例87 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物110)的制备
步骤1：将2-甲基呋喃-3-甲酸甲酯(110-a,100g,714mmol)溶于CCl4(1000mL)中，加入NBS(152.6g,857.14mmol)，AIBN(5.9g,35.71mmol)后氮气保护下置50℃下搅拌过夜。浓缩反应液，剩余物快速柱层析(PE:EA＝20:1)，得到2-(溴甲基)呋喃-3-甲酸甲酯(110-b,150g，684.93mmol，95％yield)。
步骤2：将2-(溴甲基)呋喃-3-甲酸甲酯(110-b,70g,319.63mmol)、对甲苯磺酰甘氨酸甲酯(93.3g,383.56mmol)和K2CO3(97.2g,703.20mmol)溶于乙腈(500mL)中，于30℃下搅拌过夜。将反应液过滤，浓缩反应液，剩余物快速柱层析(PE:EA＝6:1)，得到2-(((N-(2-甲氧基-2-氧代乙基)-4-甲基苯基)磺酰基)甲基)呋喃-3-甲酸甲酯(110-c,109g，285.79mmol，89％yield)。
步骤3：将2-(((N-(2-甲氧基-2-氧代乙基)-4-甲基苯基)磺酰基)甲基)呋喃-3-甲酸甲酯(110-c,4.0g,10.49mmol)溶于THF溶液中(40mL)，置于室温条件下 缓慢滴加LiHMDS(31.46mL,31.46mmol)，滴毕，室温条件下搅拌1h。向反应液中加入饱和氯化铵终止反应，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝4:1)分离纯化得4-羟基呋喃并[2,3-c]吡啶-5-甲酸甲酯(110-d,0.6g,3.11mmol,29.6％yield)。MS Calcd:193.04；MS Found:194.1([M+H]+).
步骤4：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸甲酯(110-d,0.12g,0.62mmol)和氢氧化钠(75mg,1.86mmol)依次加入4mL MeOH/H2O(4:1)混合溶液中，升温至80℃搅拌过夜。将反应液完全浓缩，向残余物中加入5N HCl溶液直至固体析出，抽滤得4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,80mg,0.45mmol,71.9％yield)。
MS Calcd:179.02；MS Found:180.02([M+H]+).
步骤5：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,26mg,0.15mmol)、DIPEA(40mg,0.31mmol)、PyBOP(77mg,0.15mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,70mg,0.12mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(110,15mg,0.02mmol,16.4％yield)。MS Calcd:726.19；MS Found:727.22([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.51(d,J＝1.2Hz,1H),8.13(d,J＝2.0Hz,1H),8.06(d,J＝8.8Hz,1H),7.96(d,J＝2.0Hz,1H),7.72(dd,J＝8.8,2.0Hz,1H),7.22(dd,J＝2.4,1.2Hz,1H),6.85-6.82(m,1H),5.32(s,2H),4.60(s,1H),4.26(q,J＝2.8Hz,2H),3.81(t,J＝5.6Hz,3H),3.54-3.48(m,2H),3.26(s,1H),3.03-2.97(m,3H),2.83–2.65(m,2H),2.53-2.51(m,2H),1.20(t,J＝7.2Hz,3H).
实施例88 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物111)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,24mg,0.13mmol)、DIPEA(36mg,0.27mmol)、PyBOP(69mg,0.13mmol)和2-(2-(3,6-二氢吡喃-4-基)-5-乙基-7-氧代-6-哌嗪-1-基-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)基)-N-(2-甲基-4-三氟甲基 苯基)乙酰胺(1-c,60mg,0.11mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(111,8mg,0.01mmol,10.2％yield)。MS Calcd:706.25；MS Found:707.31([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.51(d,J＝1.2Hz,1H),8.13(d,J＝2.0Hz,1H),7.71(d,J＝8.4Hz,1H),7.63–7.62(m,1H),7.54(dd,J＝8.8,2.0Hz,1H),7.22(dd,J＝2.4,1.2Hz,1H),6.85-6.83(m,1H),5.25(s,2H),4.60(brs,1H),4.27(q,J＝2.8Hz,2H),3.83-3.76(m,3H),3.51(t,J＝11.6Hz,2H),3.27(brs,1H),3.03-2.98(m,3H),2.83–2.65(m,2H),2.53-2.51(m,2H),2.35(s,3H),1.21(t,J＝7.2Hz,3H).
实施例89 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物112)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,23mg,0.13mmol)、DIPEA(35mg,0.27mmol)、PyBOP(66mg,0.13mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(35-c，60mg,0.11mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(112,10mg,0.01mmol,12.8％yield)。MS Calcd:724.24；MS Found:725.32([M+H]+).1H NMR(400MHz,DMSO-d6,重水交换)δ8.38(brs,1H),8.03(brs,1H),7.61–7.53(m,2H)7.14(brs,1H),6.84-6.82(m,1H),5.26(s,2H),4.61(brs,1H),4.27(q,J＝2.8Hz,2H),3.81(t,J＝5.6Hz,3H),3.53-3.46(m,2H),3.29(brs,1H),3.03-2.98(m,3H),2.85–2.65(m,2H),2.53-2.51(m,2H),2.24(s,3H),1.19(t,J＝7.2Hz,3H).
实施例90 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物113)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,47mg,0.27mmol)、DIPEA(78mg,0.6mmol)、PyBOP(150mg,0.29mmol)和(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,140mg,0.24mmol)加入DMF(3mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(113,18mg,0.02mmol,9.9％yield)。MS Calcd:740.21；MS Found:741.29([M+H]+)。1H NMR(400MHz,DMSO-d6，重水交换)δ8.38(s,1H),8.05(s,1H),8.03(s,1H),7.95(s,1H),7.71(d,J＝8.8Hz,1H),7.17(s,1H),6.83(s,1H),5.31(s,2H),4.25(d,J＝3.6Hz,2H),3.80(d,J＝5.6Hz,2H),3.56-3.49(m,2H),3.27–3.11(m,2H),2.95–2.85(m,2H),2.72–2.63(m,1H),2.53-2.51(m,2H),1.43(d,J＝6.8Hz,3H),1.22(t,J＝7.2Hz,3H).
实施例91 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物114)的制备
步骤1：将2-甲基-1H-吡咯-3-甲酸乙酯(114-a,1.52g,9.92mmol)、苯磺酰 氯(2.63g,14.88mmol)、四丁基溴化铵(0.32g,0.99mmol)溶于甲苯/H2O(10:1)混合溶液中(50mL)，置于室温条件下缓慢加入氢氧化钠(3.97g,99.22mmol)，室温条件下搅拌1h。向反应液中加入水溶液，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:6)分离纯化得2-甲基-1-(苯基磺酰基)-1H-吡咯-3-甲酸乙酯(114-b,2.3g,7.84mmol,79.3％yield)。MS Calcd:293.07；MS Found:294.2([M+H]+).
步骤2：将2-甲基-1-(苯基磺酰基)-1H-吡咯-3-甲酸乙酯(114-b,1.22g,4.16mmol)、BPO(100mg,0.42mmol)、NBS(1.11g,6.24mmol)依次加入四氯化碳溶液中(50mL)，置于80℃条件下搅拌过夜。将反应液完全浓缩，向残余物中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:5)分离纯化得2-(溴甲基)-1-(苯基磺酰基)-1H-吡咯-3-甲酸乙酯(114-c,0.8g,2.15mmol,51.7％yield)。MS Calcd:370.98；MS Found:372.0([M+H]+).
步骤3：将2-(溴甲基)-1-(苯基磺酰基)-1H-吡咯-3-甲酸乙酯(114-c,1.95g,5.24mmol)、N-(对甲苯磺酰基)甘氨酸甲酯(1.25g,5.14mmol)依次加入乙腈溶液中(10mL)，然后加入K2CO3(1.808g,13.10mmol)，置于室温条件下搅拌1h。向反应液中加入饱和氯化铵溶液，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:2)分离纯化得2-(((N-(2-甲氧基-2-氧乙基)-4-甲基苯基)磺酰胺基)甲基)-1-(苯基磺酰基)-1H-吡咯-3-甲酸乙酯(114-d,0.68g,1.27mmol,25.2％yield)。
步骤4：将2-(((N-(2-甲氧基-2-氧乙基)-4-甲基苯基)磺酰胺基)甲基)-1-(苯基磺酰基)-1H-吡咯-3-甲酸乙酯(114-d,0.68g,1.27mmol)溶于THF溶液中(5mL)，抽真空氮气置换3次，置于-78℃条件下缓慢滴加LiHMDS(3.82mL,3.82mmol)，滴毕，在此温度下继续搅拌1h。向反应液中加入饱和氯化铵终止反应，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:2)分离纯化得4-羟基-1-(苯基磺酰基)-1H-吡咯并[2,3-c]吡啶-5-甲酸甲酯(114-e,0.2g,0.6mmol,47.3％yield)。MS Calcd:332.05；MS Found:332.97([M+H]+).
步骤5：将4-羟基-1-(苯基磺酰基)-1H-吡咯并[2,3-c]吡啶-5-甲酸甲酯(114-e,0.18g,0.54mmol)和溴化苄(111mg,0.65mmol)溶于THF溶液中(5mL)，然后加入碳酸铯(0.44g,1.35mmol)，升温至70℃搅拌2h。向反应液中加入饱和氯化铵终止反应，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:2)分离纯化得4-(苄氧基)-1-(苯基磺酰基)-1H-吡咯并[2,3-c]吡啶-5-甲酸甲酯(114-f,0.21g,0.5mmol,91.7％yield)。MS Calcd:422.09；MS Found:422.93([M+H]+).
步骤6：将4-(苄氧基)-1-(苯基磺酰基)-1H-吡咯并[2,3-c]吡啶-5-甲酸甲酯(114-f,0.18g,0.43mmol)溶于2mL乙腈溶液中，然后加入甲醇钾(60mg,0.85mmol)，置于室温搅拌4h。将反应液完全浓缩得4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,120mg,0.43mmol,99.8％yield)。MS Calcd:282.10；MS Found:283.05([M+H]+)。
步骤7：将4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,47mg,0.17mmol)、DIPEA(48mg,0.38mmol)、PyBOP(94mg,0.18mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2,85mg,0.15mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，剩余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(114-h,100mg,0.12mmol,80.2％yield)。MS Calcd:829.27；MS Found:829.95([M+H]+).
步骤8：将2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(114-h,100mg,0.12mmol)溶于2mL DCM溶液中，置于-78℃条件下缓慢滴加三氯化硼(0.24mmol,0.24mL)，滴毕，缓慢升至室温搅拌过夜。加入少量甲醇终止反应，减压浓缩，残余物经prep-HPLC纯化得标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(114,8mg,0.01mmol,8.6％yield)。MS Calcd:739.22；MS Found:740.3([M+H]+)。1H NMR(400MHz,DMSO-d6，重水交换)δ8.39(s,1H),8.02(d,J＝8.4Hz,1H),7.94(d,J＝2.0Hz,1H),7.71(dd,J＝8.8,2.0Hz,1H),7.46(d,J＝3.6Hz,1H),6.83(s,1H),6.69(d,J＝2.8Hz,1H),5.30(s,2H),4.60(d,J＝12.0Hz,2H),4.25(q,J＝2.8Hz,2H),3.88(s,3H),3.80(t,J＝5.6Hz,2H),3.55–3.47(m,3H),3.02–2.98(m,3H),2.76-2.69(m,2H),2.53-2.51(m,2H),1.30–1.06(t,J＝7.2Hz,3H).
实施例92 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物115)的制备
步骤1：将4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,74mg,0.26mmol)、DIPEA(77mg,0.6mmol)、PyBOP(150mg,0.29mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(35-c，135mg,0.24mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，剩余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(115-a,120mg,0.14mmol,60.5％yield)。MS Calcd:827.32；MS Found:828.25([M+H]+)。
步骤2：将2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(115-a,120mg,0.14mmol)溶于2mL DCM溶液中，置于-78℃条件下缓慢滴加三氯化硼(0.24mmol,0.24mL)，滴毕，缓慢升至室温搅拌过夜。加入少量甲醇终止反应，减压浓缩，残余物送高压制备纯化得产物2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(3-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(115,4mg,0.005mmol,3.3％yield)。MS Calcd:737.27；MS Found:738.37([M+H]+)。
实施例93 (R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物116)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,50mg,0.28mmol)、DIEA(75mg,0.58mmol)、PyBOP(157mg,0.3mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲 基-4-(三氟甲基)苯基)乙酰胺(90-c,130mg,0.23mmol)加入DMF(3mL)，置于室温搅拌过夜。LCMS监测反应完成，向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(116,15mg,0.02mmol,8.9％yield)。MS Calcd:720.26；MS Found:721.29([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.37(s,1H),8.08(s,1H),7.70(d,J＝8.4Hz,1H),7.62(d,J＝2.0Hz,1H),7.53(dd,J＝8.4,2.0Hz,1H),7.19(s,1H),6.83(t,J＝2.0Hz,1H),5.24(s,2H),4.26(q,J＝2.8Hz,2H),3.81(t,J＝5.6Hz,3H),3.73-3.47(m,4H),3.27–3.10(m,2H),2.98–2.81(m,2H),2.71-2.62(m,1H),2.53-2.51(m,1H),2.34(s,3H),1.44(d,J＝6.8Hz,3H),1.22(t,J＝7.2Hz,3H).
实施例94 N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物117)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,36mg,0.2mmol)、DIPEA(54mg,0.42mmol)、PyBOP(113mg,0.22mmol)和N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基l)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(52-e，100mg,0.17mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物prep-HPLC纯化得标题化合物：N-(2,5-二氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(117,12mg,0.01mmol,8.3％yield)。MS Calcd:760.15；MS Found:761.17([M+H]+)。1H NMR(400MHz,DMSO-d6，重水交换)δ8.49(s,1H),8.24(d,J＝2.4Hz,1H),8.11(s,1H),8.03(s,1H),7.20(s,1H),6.82(s,1H),5.32(s,2H),4.25(m,2H),3.82-3.748(m,2H),3.53-3.47(m,3H),3.26(s,1H),3.02-2.98(m,4H),2.83(s,1H),2.65(s,1H),2.53-2.51(m,2H),1.22(t,J＝7.2Hz,3H).
实施例95 (R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物118)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,45mg,0.25mmol)、DIPEA(80mg,0.62mmol)、PyBOP(140mg,0.27mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(72-e,120mg,0.21mmol)加入DMF(3mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(118,13mg,0.02mmol,8.2％yield)。MS Calcd:738.25；MS Found:739.34([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.49(d,J＝4.8Hz,1H),8.10(d,J＝2.0Hz,1H),7.75(d,J＝12.8Hz,1H),7.65(d,J＝8.0Hz,1H),7.20(t,J＝2.8Hz,1H),6.82(t,J＝2.0Hz,1H),5.28(s,2H),4.25(q,J＝2.8Hz,2H),3.87–3.72(m,4H),3.55–3.44(m,2H),3.27–3.08(m,2H),2.94–2.82(m,2H),2.74–2.60(m,1H),2.53-2.51(m,2H),2.33(s,3H),1.44(d,J＝6.8Hz,1.5H),1.38(d,J＝6.8Hz,1.5H),1.20(t,J＝7.2Hz,3H).
实施例96 N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物119)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,37mg,0.21mmol)、DIPEA(66mg,0.51mmol)、PyBOP(116mg,0.22mmol)和N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(75-f，100mg,0.17mmol)加入DMF(3mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得到标题化合物：N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(119,13mg,0.02mmol, 10.2％yield)。MS Calcd:744.18；MS Found:745.30([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.37(s,1H),8.08(s,1H),8.05(s,1H),7.98(d,J＝7.6Hz,1H),7.18(s,1H),6.82(t,J＝2.4Hz,1H),5.34(s,2H),4.24(d,J＝2.8Hz,1H),3.80(t,J＝5.6Hz,2H),3.72-3.65(m,3H),3.52-3.48(m,2H),3.28(brs,1H),3.01-2.96(m,3H),2.82(brs,1H),2.69-2.65(m,1H),2.53-2.51(m,2H),1.18(t,J＝7.2Hz,3H).
实施例97 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物120)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,38mg,0.21mmol)、DIPEA(58mg,0.44mmol)、PyBOP(111mg,0.21mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,100mg,0.18mmol)加入DMF(3mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(120,16mg,0.02mmol,12.1％yield)。MS Calcd:724.24；MS Found:725.3([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.51(s,1H),8.12(d,J＝2.0Hz,1H),7.76(d,J＝12.8Hz,1H),7.66(d,J＝8.0Hz,1H),7.21(d,J＝2.0Hz,1H),6.83(t,J＝2.0Hz,1H),5.28(s,2H),4.26(d,J＝2.8Hz,2H),3.81(t,J＝5.6Hz,2H),3.73-3.66(m,2H),3.51(t,J＝11.2Hz,2H),3.27(brs,1H),3.02-2.96(m,3H),2.84(brs,1H),2.65(brs,1H),2.53-2.51(m,2H),2.34(s,3H),1.19(t,J＝7.2Hz,3H).
实施例98 (R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物121)的制备
步骤1：将4-羟基呋喃并[2,3-c]吡啶-5-甲酸(110-e,83mg,0.46mmol)、DIPEA (162mg,1.25mmol)、PyBOP(228mg,0.44mmol)和(R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(76-e,250mg,0.42mmol)加入DMF(3mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基呋喃并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(121,16mg,0.02mmol,12.1％yield)。MS Calcd:758.20；MS Found:759.23([M+H]+)。1H NMR(400MHz,DMSO-d6，重水交换)δ8.37(s,1H),8.09-8.05(m,2H),7.98(d,J＝7.2Hz,1H),7.19(s,1H),6.82–6.80(m,1H),5.33(s,2H),4.24(q,J＝2.8Hz,2H),3.81-3.70(m,4H),3.55-3.48(m,2H),3.27–3.07(m,2H),2.94–2.81(m,2H),2.72–2.62(m,1H),2.53-2.51(m,2H),1.44-1.37(m,3H),1.20(t,J＝7.2Hz,3H).
实施例99 N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基噻吩并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物122)的制备
步骤1：将噻吩-3-甲酸(122-a,5.0g,39.03mmol)溶于THF溶液中(50mL)，置于-78℃条件下缓慢加入TMEDA(14.54mL,97.58mmol)，在此温度下搅拌10min，然后缓慢滴加正丁基锂(39.03mL,97.58mol)，滴毕，在此温度下继续搅拌1h，最后加入DMF(6.68mL,85.87mol)，继续搅拌30min。向反应液中加入稀盐酸终止反应，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得粗品2-甲酰基噻吩-3-甲酸(122-b,5.8g,37.13mmol,95.1％yield)。MS Calcd:155.99；MS Found:155.210([M-H]-).
步骤2：将2-甲酰基噻吩-3-甲酸(122-b,5.8g,37.13mmol)和碳酸钾(12.03g,87.04mmol)依次加入DMF溶液中(50mL)，然后加入碘乙烷(6.78g,43.52mmol)，置于50℃条件下搅拌3h。将反应液完全浓缩，向残余物中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:5)分离纯化得2-甲酰基噻吩-3-甲酸乙酯(122-c,3.56g,19.33mmol, 52.1％yield)。MS Calcd:184.02；MS Found:185.0([M+H]+).
步骤3：将2-甲酰基噻吩-3-甲酸乙酯(122-c,3.08g,16.72mmol)和溶于THF/MeOH(7:1)混合溶液中(25mL)，置于0℃条件下分批加入硼氢化钠(0.95g,25.08mmol)，升至室温搅10min。将反应液完全浓缩，向残余物中加入饱和氯化铵溶液，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:5)分离纯化得2-(羟甲基)噻吩-3-甲酸乙酯(122-d,2.5g,13.43mmol,80.3％yield)。MS Calcd:186.04；MS Found:169.0([M-OH]+).
步骤4：将2-(羟甲基)噻吩-3-甲酸乙酯(122-d,2.2g,11.82mmol)加入DCM溶液中(20mL)，置于0℃条件下分批加入三溴化磷(1.28g,4.73mmol)，升至室温搅拌30min。向残余物中加入水，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:5)分离纯化得2-(溴甲基)噻吩-3-甲酸乙酯(122-e,2.5g,10.04mmol,84.9％yield)。
步骤5：将2-(溴甲基)噻吩-3-甲酸乙酯(122-e,2.09g,8.39mmol)、N-(对甲苯磺酰基)甘氨酸甲酯(2.45g,10.07mmol)和碳酸钾(2.55g,18.46mmol)依次加入乙腈溶液中(30mL)，置于室温条件下搅拌过夜。向反应液中加入饱和氯化铵溶液，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:2)分离纯化得2-(((N-(2-甲氧基-2-氧乙基)-4-甲基苯基)磺酰胺基)甲基)噻吩-3-甲酸乙酯(122-f,3.2g,7.78mmol,92.7％yield)。MS Calcd:411.08；MS Found:412.2([M+H]+).
步骤6：将2-(((N-(2-甲氧基-2-氧乙基)-4-甲基苯基)磺酰胺基)甲基)噻吩-3-甲酸乙酯(122-f,2.0g,4.86mmol)溶于THF溶液中(20mL)，抽真空氮气置换3次，置于-78℃条件下缓慢滴加LiHMDS(19.44mL,19.44mmol)，滴毕，在此温度下继续搅拌1h。向反应液中加入饱和氯化铵终止反应，乙酸乙酯萃取，饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经硅胶柱层析(EA:PE＝1:2)分离纯化得4-羟基噻吩并[2,3-c]吡啶-5-甲酸甲酯(122-g,0.7g,3.35mmol,68.8％yield)。MS Calcd:209.01；MS Found:210.10([M+H]+).
步骤7：将4-羟基噻吩并[2,3-c]吡啶-5-甲酸甲酯(122-g,0.2g,0.96mmol)和氢氧化钠(115mg,2.87mmol)依次加入4mL MeOH/H2O(4:1)混合溶液中，升温至80℃搅拌过夜。TLC监测反应完成，将反应液完全浓缩，向残余物中加入5N HCl溶液直至固体析出，抽滤得4-羟基噻吩并[2,3-c]吡啶-5-甲酸(122-h,100mg,0.51mmol,53.6％yield)。MS Calcd:195.00；MS Found:196.08([M+H]+).
步骤8：将4-羟基噻吩并[2,3-c]吡啶-5-甲酸(122-h,42mg,0.21mmol)、DIEA(68mg,0.53mmol)、PyBOP(120mg,0.23mmol)和N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(int-2，100mg,0.18mmol)加入DMF(2mL)，置于室温搅拌过 夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化的标题化合物：N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基噻吩并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(122,14mg,0.02mmol,10.6％yield)。MS Calcd:742.17；MS Found:743.24([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.76(s,1H),8.04-8.01(m,2H),7.94(d,J＝2.0Hz,1H),7.73–7.69(m,2H),6.84-6.82(m,1H),5.31(s,2H),4.60(brs,1H),4.25(q,J＝2.8Hz,2H),4.07(brs,1H),3.80(t,J＝5.6Hz,2H),3.56–3.49(m,2H),3.30(brs,1H),3.02-2.96(m,3H),2.84-2.68(m,2H),2.53-2.51(m,2H),1.20(t,J＝7.2Hz,3H).
实施例100 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基噻吩并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物123)的制备
步骤1：将4-羟基噻吩并[2,3-c]吡啶-5-甲酸(122-h,43mg,0.22mmol)、DIPEA(71mg,0.55mmol)、PyBOP(124mg,0.24mmol)和2-(2-(3,6-二氢吡喃-4-基)-5-乙基-7-氧代-6-哌嗪-1-基-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)基)-N-(2-甲基-4-三氟甲基苯基)乙酰胺(1-c,100mg,0.18mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经prep-HPLC纯化的标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基噻吩并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(123,15mg,0.02mmol,10.4％yield)。MS Calcd:722.22；MS Found:723.26([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.36(s,1H),8.00(brs,2H),7.71(t,J＝7.2Hz,1H),7.62(d,J＝2.0Hz,1H),7.53(dd,J＝8.8,2.0Hz,1H),6.84-6.82(m,1H),5.25(s,2H),4.61(brs,1H),4.26(q,J＝2.8Hz,2H),4.05(brs,1H),3.81(t,J＝5.6Hz,2H),3.56–3.50(m,2H),3.31(brs,1H),3.03-2.96(m,3H),2.83–2.68(m,2H),2.53-2.51(m,2H),2.34(s,3H),1.21(t,J＝7.2Hz,3H).
实施例101 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物124)的制备
步骤1：将4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,100mg,0.35mmol)、DIPEA(103mg,0.8mmol)、PyBOP(200mg,0.38mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(19-c,180mg,0.32mmol)加入DMF(4mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(124-a,140mg,0.17mmol,53.0％yield)。MS Calcd:827.32；MS Found:828.41([M+H]+).
步骤2：将2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(124-a,140mg,0.17mmol)溶于2mL DCM溶液中，置于-78℃条件下缓慢滴加三氯化硼(0.34mmol,0.34mL)，滴毕，缓慢升至室温搅拌过夜。加入少量饱和氯化铵溶液终止反应，二氯甲烷萃取，减压浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(5-氟-2-甲基-4-(三氟甲基)苯基)乙酰胺(124,14mg,0.02mmol,11.0％yield)。MS Calcd:737.27；MS Found:738.27([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.40(s,1H),7.75(d,J＝12.8Hz,1H),7.65(d,J＝8.0Hz,1H),7.46(d,J＝3.2Hz,1H),6.82(s,1H),6.69(d,J＝3.2Hz,1H),5.28(s,2H),4.63(d,J＝12.4Hz,2H),4.25(d,J＝3.2Hz,2H),3.89(s,3H),3.80(t,J＝5.6Hz,2H),3.56-5.50(m,2H),3.16–2.98(m,4H),2.76(brs,2H),2.53-2.51(m,2H),2.33(s,3H),1.20(t,J＝7.2Hz,3H).
实施例102 (R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物125)的制备
步骤1：将4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,100mg,0.35mmol)、DIPEA(103mg,0.8mmol)、PyBOP(200mg,0.38mmol)和(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(90-c,180mg,0.32mmol)加入DMF(4mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得(R)-2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(125-a,160mg,0.19mmol,60.4％yield)。MS Calcd:823.34；MS Found:824.35([M+H]+).
步骤2：将(R)-2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(125-a,160mg,0.19mmol)溶于2mL DCM溶液中，置于-78℃条件下缓慢滴加三氯化硼(0.83mmol,0.83mL)，滴毕，缓慢升至室温搅拌过夜。加入少量饱和氯化铵溶液终止反应，二氯甲烷萃取，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(125,16mg,0.02mmol,9.7％yield)。MS Calcd:733.29；MS Found:734.3([M+H]+).1H NMR(400MHz,DMSO-d6，重水交换)δ8.39(s,1H),7.69(d,J＝8.4Hz,1H),7.62(d,J＝2.4Hz,1H),7.53(dd,J＝8.4,2.4Hz,1H),7.46(d,J＝3.2Hz,1H),6.84-6.82(m,1H),6.69(d,J＝3.2Hz,1H),5.24(s,2H),4.26(q,J＝2.8Hz,2H),3.88(s,3H),3.81(t,J＝5.6Hz,2H),3.74-3.70(m,2H),3.57-3.51(m,2H),3.28–3.14(m,2H),3.00–2.81(m,2H),2.72–2.66(m,2H),2.53-2.51(m,2H),2.34(s,3H),1.45(d,J＝7.2Hz,3H),1.23(t,J＝7.2Hz,3H).
实施例103 (R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物126)的制备
步骤1：将4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,100mg,0.28mmol)、DIPEA(84mg,0.65mmol)、PyBOP(162mg,0.31mmol)和(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(14-b,150mg,0.26mmol)加入DMF(2mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得(R)-2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(126-a,160mg,0.19mmol,73.3％yield)。MS Calcd:843.29；MS Found:844.21([M+H]+).
步骤2：将(R)-2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-4-(三氟甲基)苯基)乙酰胺(126-a,110mg,0.13mmol)溶于2mL DCM溶液中，置于-78℃条件下缓慢滴加三氯化硼(0.52mmol,0.52mL)，滴毕，缓慢升至室温搅拌过夜。加入少量甲醇终止反应，减压浓缩，残余物经prep-HPLC纯化得标题化合物：(R)-N-(2-氯-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)-3-甲基哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(126,8mg,0.01mmol,8.1％yield)。MS Calcd:753.24；MS Found:754.25([M+H]+).1H NMR(400MHz,DMSO-d6,重水交换)δ8.39(s,1H),8.04(d,J＝8.8Hz,1H),7.94(d,J＝2.4Hz,1H),7.71(dd,J＝8.8,2.4Hz,1H),7.46(d,J＝3.2Hz,1H),6.84–6.82(m,1H),6.69(d,J＝3.2Hz,1H),5.31(s,2H),4.86(brs,0.5H),4.66(brs,0.5H),4.50–4.44(m,0.5H),4.32-4.21(m,2.5H),3.88(s,3H),3.80(t,J＝5.6Hz,2H),3.75-3.69(m,1.5H),3.57-3.52(m,1.5H),3.27–3.13(m,1.5H),2.93–2.81(m,1.5H),2.71–2.65(m,1H),2.53-2.51(m,2H),1.44(t,J＝5.6Hz,3H),1.22(t,J＝7.2Hz,3H).
实施例104 2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(化合物127)的制备
步骤1：将4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,131mg,0.46mmol)、DIPEA(136mg,1.05mmol)、PyBOP(263mg,0.51mmol)和2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(1-c，230mg,0.42mmol)加入DMF(4mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(127-a,180mg,0.22mmol,52.7％yield)。MS Calcd:809.33；MS Found:810.04([M+H]+).
步骤2：将2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(127-a,800mg,0.1mmol)溶于2mL DCM溶液中，置于-78℃条件下缓慢滴加三氯化硼(0.4mmol,0.4mL)，滴毕，缓慢升至室温搅拌过夜。加入少量甲醇终止反应，减压浓缩，残余物经prep-HPLC纯化得标题化合物：2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-甲基-4-(三氟甲基)苯基)乙酰胺(127,16mg,0.02mmol,22.1％yield)。MS Calcd:719.28；MS Found:720.3([M+H]+).1H NMR(400MHz,DMSO-d6,重水交换)δ8.42(s,1H),7.72(d,J＝8.4Hz,1H),7.63(d,J＝2.0Hz,1H),7.54(dd,J＝8.4,2.0Hz,1H),7.48(d,J＝2.8Hz,1H),6.85-6.83(m,1H),6.70(d,J＝2.8Hz,1H),5.26(s,2H),4.69(brs,2H),4.27(q,J＝2.8Hz,2H),3.90(s,3H),3.82(t,J＝5.6Hz,2H),3.61-3.51(m,2H),3.32-3.00(m,4H),2.77(brs,2H),2.53-2.51(m,2H),2.36(s,3H),1.23(t,J＝7.2Hz,3H).
实施例105 N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(化合物128)的制备
步骤1：将4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-甲酸(114-g,106mg,0.38mmol)、DIPEA(110mg,0.86mmol)、PyBOP(214mg,0.41mmol)和N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-6-(哌嗪-1-基)-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(75-f，200mg,0.34mmol)加入DMF(4mL)，置于室温搅拌过夜。向反应液中加入水，DCM萃取，饱和食盐水洗涤，减压浓缩，残余物经硅胶柱层析(DCM:MeOH＝20:1)分离纯化得2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-5-氟-4-(三氟甲基)苯基)乙酰胺(128-a,120mg,0.14mmol,41.3％yield)。MS Calcd:847.26；MS Found:847.92([M+H]+).
步骤2：将2-(6-(4-(4-(苄氧基)-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)-N-(2-氯-5-氟-4-(三氟甲基)苯基)乙酰胺(128-a,120mg,0.14mmol)溶于2mL DCM溶液中，置于-78℃条件下缓慢滴加三氯化硼(0.34mmol,0.34mL)，滴毕，缓慢升至室温搅拌过夜。加入少量甲醇终止反应，减压浓缩，残余物经prep-HPLC纯化得到标题化合物：N-(2-氯-5-氟-4-(三氟甲基)苯基)-2-(2-(3,6-二氢-2H-吡喃-4-基)-5-乙基-6-(4-(4-羟基-1-甲基-1H-吡咯并[2,3-c]吡啶-5-羰基)哌嗪-1-基)-7-氧代-[1,2,4]三唑并[1,5-a]嘧啶-4(7H)-基)乙酰胺(128,18mg,0.02mmol,15.9％yield)。MS Calcd:757.22；MS Found:758.21([M+H]+).1H NMR(400MHz,DMSO-d6,重水交换)δ8.41(s,1H),8.09(d,J＝12.8Hz,1H),7.99(d,J＝7.2Hz,1H),7.48(d,J＝3.2Hz,1H),6.83-6.80(m,1H),6.69(d,J＝3.2Hz,1H),5.35(s,2H),4.69(brs,2H),4.25(q,J＝2.8Hz,2H),3.90(s,3H),3.80(t,J＝5.6Hz,2H),3.56-3.50(m,2H),3.18–2.97(m,4H),2.77(brs,2H),2.53-2.51(m,2H),1.20(t,J＝7.2Hz,3H).
实施例106 体外酶学活性评价：
通过测定IC50值来评价化合物对人的WRN解旋酶的抑制活性。
主要试剂及耗材：

实验方案：将两条单链DNA Oligo退火形成dsDNA作为WRN(aa500-946)酶学反应的底物。反应缓冲液成分及浓度：5mM NaCl、25mM Tris base(pH 8.0)、0.01％Tween20、2mM MgCl2、1mM TCEP、0.02％BSA。实验反应均在微孔板恒温振荡箱中完成，振荡箱参数设置为25℃，300rpm。实验采用5μL反应体系，实验起始，首先在384孔板中，化合物组加入2μL WRN(aa500-946)溶液(终浓度1.2nM)和1μL化合物溶液(化合物最大终浓度10μM，3×梯度稀释，10个浓度点，DMSO终浓度1％)，最大信号组加入2μL WRN(aa500-946)溶液(终浓度1.2nM)和1μL反应缓冲液(DMSO终浓度1％)，最小信号组加入2μL反应缓冲液和1μL反应缓冲液(DMSO终浓度1％)，各组孵育30min，随后加入2μL底物溶液(含终浓度0.1nM的dsDNA和10μM的ATP)反应60min，随后再加入5μL ADP Glo Reagent孵育40min，最后再加入10μL Kinase Detection Reagent孵育30min，孵育完成后通过Tecan Spark读取Luminescence。抑制率％＝{1-[(化合物组发光信号-最小信号组)/(最大信号组-最小信号组)]}*100％。数据分析使用软件GraphPad Prism 8，采用“log(inhibitor)vs.response--Variable slope(four  parameters)”方法拟合数据，得到化合物的IC50值。
表1.本发明化合物对WRN的半数抑制浓度IC50

结论：结果显示，本发明化合物对WRN有良好的抑制活性。
实施例107 细胞活性及选择性测试实验:
通过测试WRN非共价抑制剂对MSI-H细胞株SW48和MSS细胞株SW620的增殖抑制活性以评价抑制剂的细胞活性和选择性。
主要试剂及耗材：
将SW48和SW620细胞培养至生长密度80％左右时，用Trypsin消化细胞并计数，用SW48细胞相应培养液DMEM和SW620细胞相应培养液DMEM-F12 稀释细胞至1*10^4个/mL，然后于96孔板中加入100μL稀释好的细胞悬液，使各孔内细胞数目为1000个，随后置于二氧化碳培养箱(37℃，5％CO2)培养过夜至细胞贴壁。将待测化合物用DMSO溶解并稀释至5mM，随后用自动化微孔板移液分液系统在96孔板(V型底)中用DMSO将化合物从5mM浓度起始3倍梯度稀释8次至0.0008mM(共9个浓度点)，然后再用SW48和SW620细胞相应的培养液将梯度稀释后的化合物分别稀释500倍，至DMSO终浓度为0.2％，化合物终浓度为10μM～0.0015μM。孔内仅含0.2％DMSO的相应培养液为空白组。用移液器彻底吸去SW48和SW620细胞培养板孔内的培养液，随后于孔内，化合物组加入100μL含相应浓度的化合物的培养液，DMSO对照组加入仅含0.2％DMSO的培养液。空白组、化合物组和DMSO对照组于二氧化碳培养箱培养4天。4天后，将培养板从培养箱中取出置于室温使其温度降至室温，待恢复至室温后，于各组孔内加入50μL配制好的CellTiter-Glo溶液，随后置于微孔板恒温振荡箱25℃，800rpm孵育10min。孵育完成后，用移液器吸取各孔内100μL反应溶液转移至黑色96孔板中，于酶标仪读取Luminescence。
每个浓度的化合物的抑制百分数(％)是根据每个检测板中包含的最小荧光信号组(空白组)和最大荧光信号组(DMSO对照组)的信号计算的。其中最小荧光信号组无细胞，仅含SW48和SW620细胞培养液(0.2％DMSO)，最大荧光信号组无化合物，仅含SW48和SW620细胞(0.2％DMSO)。各个化合物浓度的抑制率计算方式如下：抑制率％＝{1-[(化合物组发光信号-最小荧光信号组)/(最大发光信号组-最小发光信号组)]}*100％，使用GraphPad Prism软件获得GI50值和非线性回归曲线拟合，化合物细胞选择性＝SW620GI50/SW48GI50，GI50及选择性值见下表。
表2本发明化合物SW48和SW620细胞增殖抑制活性及选择性

结论：结果显示，本发明化合物对MSI-H细胞SW48具有较强的增殖抑制活性，对于MSS细胞SW620增殖抑制则很弱，表明本发明化合物具有良好的选择性。
实施例108 药代动力学实验：
实验材料、方法及结果分析：
实验动物为健康成年BALB/c雌性小鼠(由北京华阜康生物科技股份有限公司提供)；
小鼠给药方式及样品采集：
对BALB/c雌性小鼠单次灌胃给药(10mg/kg，溶媒：5％DMSO+5％solutol+90％saline)(DMSO，成都市科隆化学品有限公司；Solutol，BASF，42966-1KG；Saline，四川美大康佳乐药业有限公司，100mL_0.9g)，分别于给药后0.25，0.5，1，2，4，6，8h收集小鼠不同时间点眼底静脉丛全血60μL，4000rpm离心10min取血浆。
样品分析：
取小鼠血浆样品10μL，分别加入190μL含内标的乙腈溶液沉淀蛋白，涡旋10min，随后4000rpm离心10min，取上清液于96孔板中。放入LC-MS/MS(质谱仪，TQ4500，液相色谱仪，岛津30AD)进行分析。
应用LC-MS/MS法测定小鼠灌胃给予实施例化合物后不同时刻血浆中的药物浓度，并计算相关药动学参数，研究化合物在小鼠体内的药代动力学行为，评价其药动学特征。
对照例1参照专利WO2022249060A1中example 58制备而成，结构如下：
表3.本发明化合物在小鼠体内的药代动力学参数

结论：结果显示，本发明化合物在小鼠体内展现出良好的药代动力学性质；与对照例1相比，本发明化合物在血浆中暴露量AUC和Cmax均有显著提高。
实施例109 小鼠瘤组织分布实验：
实验材料、方法及结果分析：
实验动物为健康成年Balb/c nude雌性小鼠(由四川维通利华实验动物技术有限公司提供)；
给药方式及样品采集：
在Balb/c nude雌性小鼠右侧腋下皮下接种SW48细胞(南京科佰生物科技有限公司)，每只小鼠接种0.1mL，接种量为3×106个细胞。当动物平均肿瘤体积达到194mm3时，对Balb/c nude雌性小鼠灌胃给药(10mg/kg，10％DMSO+10％Solutol+80％H2O)(DMSO:AR 500ml，科隆；Solutol：42966-1KG，BASF)，于给药后1h和4h收集小鼠不同时间点血浆和肿瘤样品。其中收集眼底静脉丛全血60μL，4000rpm离心6min取血浆；动物安乐死后取肿瘤组织，滤纸擦净水分，称量并按比例加入一定量生理盐水，匀浆后装于EP管中。
样品分析：
取小鼠血浆和肿瘤样品10μL，分别加入一定体积含内标的乙腈溶液沉淀蛋白，涡旋10min，随后4000rpm离心10min，取上清液于96孔板中。放入LC-MS/MS(Shimadzu LC-30A，AB API 4500)进行分析。
应用LC-MS/MS法测定小鼠灌胃给予实施例化合物后不同时刻血浆和肿瘤中的药物浓度，并计算相关药动学参数，研究化合物在小鼠体内的药代动力学行为，评价其药动学特征。
表4
结论：结果显示，本发明化合物在荷瘤小鼠肿瘤组织中有较好的分布。
实施例110 hERG抑制研究
实验操作：
采用PredictorTMhERG Fluorescence Polarization Assay Kit(ThermoFisher Scientific PV5365)进行hERG抑制检测。预先配制4×PredictorTMhERG Tracer Red和4×E-4031阳性对照。在384孔板中，Assay Blank孔中加入5μL PredictororhERG FP Assay Buffer和5μL PredictorTMhERG Membrane。阴性对照、阳性对照、检测化合物孔中分别加入2.5μL PredictorTMhERG FP Assay Buffer、2.5μL 4×E-4031 Positive Control、2.5μL 4×待测化合物，再依次加入5μL PredictorTM hERG Membrane及2.5μL 4×PredictorTMhERG Tracer Red。在室温(25℃)下避光孵育3.5h。反应结束后，于酶标仪(Tecan，)检测荧光偏振值，G Factor为2.14。
抑制率计算：
应用如下公式计算化合物或阳性对照的抑制率。
抑制率％＝(mP阴性对照-mP化合物)/(mP阴性对照-mP阳性对照)*100％
表5.hERG抑制试验结果
实施例111 人肝微粒体稳定性实验:
实验材料、方法及结果分析：
实验所用为人肝微粒体(供应商：瑞德肝脏疾病研究(上海)有限公司，品牌：BIOIVT，货号：X008070)；NADPH(还原型烟酰胺腺嘌呤二核苷酸磷酸，货号：481973-500mg，sigma)。
试剂配制：
PBS：0.1M KH2PO4和K2HPO4缓冲液，pH 7.4。
MgCl2：称取一定量的MgCl2，用PBS配制成16mM的MgCl2溶液。
NADPH：称取一定量的NADPH，用16mM的MgCl2溶液将NADPH配制到4mM，最终孵育浓度为1mM。
化合物：用PBS将待测化合物配制到4μΜ，最终孵育浓度为1μM。
肝微粒体：用PBS将肝微粒体稀释到1mg/mL，最终孵育浓度为0.5mg/mL。
实验步骤：
试管中加入待测化合物，随后加入配好的NADPH，混合均匀。置于37℃，220rpm恒温箱预孵，预孵5min后加入肝微粒体起始反应，复孔操作。
在0min、5min、15min、30min、60min时，对应管中加入一定体积含内标的冰乙腈溶液沉淀蛋白，振荡涡旋5min，随后4000rpm离心10min，取上清液于96孔板中。放入LC-MS/MS进行分析。
通过LC-MS/MS测定实施例化合物的浓度(峰面积比)，在Excel中以“Ln(化合物残余量％)”对“孵育时间”作图获得速率常数，从而计算药物的半衰期与内在清除率，为体内清除率的预测提供依据。
数据分析：
CLint＝(0.693/t1/2，微粒体)×[孵化液体积(mL)/微粒体蛋白质量(mg)]
×[微粒体蛋白质量(mg)/肝质量(g)]×[肝质量(g)/体重(kg)][1]
CLH＝CLint×fu×Qh/(CLint×fu+Qh)
式中
CLint--固有清除率(ml/min/kg)
CLH--肝清除率(ml/min/kg)
fu--血浆蛋白非结合分数为1
Qh--肝血流量
表6.人肝微粒稳定性结果
[1]Davies B,Morris T.Physiological parameters in laboratory animals and humans.Pharm  Res.1993；10:1093-5.
结论：结果显示，本发明化合物在人肝微粒体中具有良好的稳定性。
产业上的可应用性
本发明的化合物具有优异的WRN抑制活性，可成为治疗或预防与该作用相关的疾病的药物。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。

Claims (19)

  1. 一种式(I)所示的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐：
    其中，
    R1选自卤素、C1-6烷基、C2-6烯基、C2-6炔基、5元或6元环烯基、5-10元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所述烷基、烯基、炔基、环烯基、杂环基、杂芳基、苯基各自独立地是未取代的或被1个或多个相同或不同的Rg取代；
    每个Rg各自独立地选自：羟基、卤素、C1-4烷基、C1-4烷氧基、HOC(O)-(CH2)n-、H3C-C(O)(CH2)n-、C1-4烷基-O-C(O)(CH2)n-、＝O、氮杂环丁基、吡咯烷基、R’R”N-，其中，R’、R”各自独立地选自H、C1-4烷基、C1-4卤代烷基，n＝0，1或2；所述氮杂环丁基或吡咯烷基通过N原子连接到R1主基团上；所述C1-4烷基、C1-4烷氧基、氮杂环丁基、吡咯烷基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
    R2选自 其中，L选自7-10元螺杂环烷基、所述螺杂环烷基含有2-3个N原子；
    R3选自环丙基、甲氧基、-SCH3、C1-4烷基，所述烷基是未取代的或被1个或多个选自羟基或卤素的取代基取代；
    R4选自H、卤素、羟基、氰基、C1-4烷基、C3-5环烷基、C1-4烷氧基，所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
    R5选自H、甲氧基、羟基、氰基、甲基、卤素；
    R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个或多个卤素取代；
    R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基、C1-4烷氧基；所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取 代；
    X选自N或CR8，R8为H或卤素；
    R2a选自6-14元芳基、5-14元杂芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述芳基、杂芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rc取代；
    R2b选自5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环烷基、5-14元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个选自N、O或S的杂原子；所述5-14元杂环烷基、5-14元杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环烷基、5-14元杂环烯基各自独立地被一个或多个Rd取代；
    R2e选自5-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述杂芳基、芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rf取代；
    每个Rc各自独立地选自卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基、氰基、氨基；
    每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、C1-6卤代烷氧基、C3-6卤代环烷基、羟基、卤素、4-7元杂环烷基、5元或6元杂芳基、苯基；所述4-7元杂环烷基含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代；
    每个Rf各自独立地选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基。
  2. 式(I)所示的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    其中，
    R1选自卤素、C1-6烷基、C2-6烯基、C2-6炔基、5元或6元环烯基、5元或6元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基含有1-2个选自N、O或S的杂原子，所述烷基、烯基、炔基、环烯基、杂环基、杂芳基、苯基是未取代的或被1个或多个相同或不同的Rg取代；
    每个Rg各自独立地选自：羟基、卤素、C1-4烷基、C1-4烷氧基、HOC(O)-(CH2)n-、H3C-C(O)(CH2)n-、C1-4烷基-O-C(O)(CH2)n-、＝O、氮杂环丁基、吡咯烷基、R’R”N-，其中，R’、R”各自独立地选自H、C1-4烷基、C1-4卤代烷基，n＝0，1或2；所述氮杂环丁基或吡咯烷基通过N原子连接到R1主基团上；所述C1-4烷基、C1-4烷氧基、氮杂环丁基、吡咯烷基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
    R2选自其中，L选自7-10元螺杂环基、
    R3选自环丙基、甲氧基、-SCH3、C1-4烷基，所述烷基是未取代的或被1个或多个选自羟基或卤素的取代基取代；
    R4选自H、卤素、羟基、氰基、C1-4烷基、C3-5环烷基、C1-4烷氧基，所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
    R5选自H、甲氧基、羟基、氰基、甲基、卤素；
    R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个或多个卤素取代；
    R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基、C1-4烷氧基；所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
    X选自N或CR8，R8为H或卤素；
    R2a选自6-14元芳基、5-14元杂芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述芳基、杂芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rc取代；
    R2b选自5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基各自独立地被一个或多个Rd取代；
    R2e选自5-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述杂芳基、芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rf取代；
    每个Rc各自独立地选自卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基、氰基、氨基；
    每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、C1-6卤代烷氧基、C3-6卤代环烷基、羟基、4-7元杂环烷基或卤素；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；
    每个Rf各自独立地选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基。
  3. 式(I)所示的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    其中，
    R1选自卤素、C1-6烷基、C2-6烯基、C2-6炔基、5元或6元环烯基、5元或6元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基含有1-2个选自N、O或S的杂原子，所述烷基、烯基、炔基、环烯基、杂环基、杂芳基、苯基是未取代的或被1个或多个相同或不同的Rg取代；
    每个Rg各自独立地选自：羟基、卤素、C1-4烷基、C1-4烷氧基、HOC(O)-(CH2)n-、H3C-C(O)(CH2)n-、C1-4烷基-O-C(O)(CH2)n-、＝O、氮杂环丁基、吡咯烷基、R’R”N-，其中，R’、R”各自独立地选自H、C1-4烷基、C1-4卤代烷基，n＝0，1或2；所述氮杂环丁基或吡咯烷基通过N原子连接到R1主基团上；所述C1-4烷基、C1-4烷氧基、氮杂环丁基、吡咯烷基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
    R2选自其中，L选自7-10元螺杂环基、
    R3选自环丙基、甲氧基、-SCH3、C1-4烷基，所述烷基是未取代的或被1个或多个选自羟基或卤素的取代基取代；
    R4选自H、卤素、羟基、氰基、C1-4烷基、C3-5环烷基、C1-4烷氧基，所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
    R5选自H、甲氧基、羟基、氰基、甲基、卤素；
    R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个或多个卤素取代；
    R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基、C1-4烷氧基；所述烷基、环烷基、烷氧基是未取代的或被1个或多个卤素取代；
    X选自N或CR8，R8为H或卤素；
    R2a选自6-14元芳基、5-14元杂芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述芳基、杂芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rc取代；
    R2b选自5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-14元杂芳基、6-14元芳基、5-14元杂环基各自独立地被一个或多个Rd取代；
    R2e选自5-14元杂芳基、6-14元芳基、5-14元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述杂芳基、芳基、杂环基是未取代的或各自独立地被一个或多个相同或不同的Rf取代；
    每个Rc各自独立地选自卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基、氰基、氨基；
    每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、C1-6卤代烷氧基、C3-6卤代环烷基、羟基或卤素；
    每个Rf各自独立地选自羟基、氰基、氨基、卤素、C1-6烷基、C1-6卤代烷基、C1-6烷氧基、C1-6卤代烷氧基、C3-6环烷基、C3-6卤代环烷基。
  4. 如权利要求1-3任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R2选自
    优选的，R2选自
    优选的，R2选自
    R2b选自5元杂芳基、8-14元杂芳基；所述5元杂芳基、8-14元杂芳基各自独立地含有1-4个选自N、O和S的杂原子；所述5元杂芳基、8-14元杂芳基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、8元杂芳基、9元杂芳基、10元杂芳基；所 述5元杂芳基、8元杂芳基、9元杂芳基、10元杂芳基各自独立地含有1-4个选自N、O和S的杂原子；所述5元杂芳基、8元杂芳基、9元杂芳基、10元杂芳基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、5元杂芳基并5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基、6元杂芳基并6元杂芳基；所述5元杂芳基、6元杂芳基各自独立地含有1-2个选自N、O和S的杂原子；所述5元杂芳基、5元杂芳基并5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基、6元杂芳基并6元杂芳基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基；所述5元杂芳基、6元杂芳基各自独立地含有1-2个选自N、O和S的杂原子；所述5元杂芳基、苯并5元杂芳基、6元杂芳基并5元杂芳基、苯并6元杂芳基各自独立地被一个或多个Rd取代；
    优选的，R2b选自吡咯基、呋喃基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基、苯并吡咯基、苯并呋喃基、苯并噻吩基、苯并吡唑基、苯并咪唑基、苯并噻唑基、苯并异噻唑基、苯并噁唑基、苯并异噁唑基、吡啶并吡咯基、吡啶并呋喃基、吡啶并噻吩基、吡啶并吡唑基、吡啶并咪唑基、吡啶并噻唑基、吡啶并异噻唑基、吡啶并噁唑基、吡啶并异噁唑基、苯并吡啶基、苯并嘧啶基；所述吡咯基、呋喃基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基、苯并吡咯基、苯并呋喃基、苯并噻吩基、苯并吡唑基、苯并咪唑基、苯并噻唑基、苯并异噻唑基、苯并噁唑基、苯并异噁唑基、吡啶并吡咯基、吡啶并呋喃基、吡啶并噻吩基、吡啶并吡唑基、吡啶并咪唑基、吡啶并噻唑基、吡啶并异噻唑基、吡啶并噁唑基、吡啶并异噁唑基、苯并吡啶基、苯并嘧啶基各自独立地被一个或多个Rd取代；
    优选的，R2b选自吡唑基、苯并吡啶基、异噁唑基、异噻唑基、吡啶并吡唑基、吡啶并咪唑基、吡啶并呋喃基、吡啶并吡咯基、吡啶并噻吩基；所述吡唑基、苯并吡啶基、异噁唑基、异噻唑基、吡啶并吡唑基、吡啶并咪唑基、吡啶并呋喃基、吡啶并吡咯基、吡啶并噻吩基各自独立地被一个或多个Rd取代；
    优选的，R2b选自 所述 各自独立地被一个或多个Rd取代；
    优选的，R2b选自
    优选的，R2b选自
    每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、羟基、卤素，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1或2个选自N、O和S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O和S的杂原子；
    优选的，每个Rd各自独立地选自C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、羟基、卤素，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1或2个选自N、O和S的杂原子；所述5元或6元杂芳基各自独立地含有1个、2个或3个选自N、O和S的杂原子；
    优选的，每个Rd各自独立地选自C1-4烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C1-4烷氧基、4-7元杂环烷基、6元杂芳基、苯基、羟基、卤素，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1个或2个O杂原子；所述6元杂芳基含有1个或2个N杂原子；
    优选的，每个Rd各自独立地选自C1-2烷基、C3-4烷基、C3-4环烷基、C5-6环烷基、C1-2氟代烷基、C3-4氟代烷基、C1-2烷氧基、C3-4烷氧基、4-7元杂环烷基、吡啶基、嘧啶基、哒嗪基、吡嗪基、苯基、羟基、卤素，其中有且只有一个Rd为 羟基；所述4-7元杂环烷基含有1个O杂原子；
    优选的，每个Rd各自独立地选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基、环丙基、环丁基、环戊基、环己基、氟代甲基、氟代乙基、氟代丙基、氟代异丙基、甲氧基、乙氧基、4-7元杂环烷基、吡啶基、羟基、F、Cl、Br、苯基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基含有1个O杂原子；
    优选的，每个Rd各自独立地选自甲基、异丙基、叔丁基、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、 Cl、苯基、羟基，其中有且只有一个Rd为羟基；
    优选的，所述羟基取代在R2b与主基团连接位点的邻位或间位的碳原子上；
    优选的，所述羟基取代在R2b与主基团连接位点的邻位的碳原子上；
    最优选的，R2b选自
  5. 如权利要求1-3任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R2选自
    优选的，R2b选自5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环烷基、5-10元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个 选自N、O或S的杂原子；所述5-10元杂环烷基、5-10元杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环烷基、5-10元杂环烯基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个选自N、O或S的杂原子；所述杂环烷基、杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、8-10元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基，所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-4个选自N、O或S的杂原子；所述杂环烷基、杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述5元杂芳基、8-10元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、8-10元杂环烷基、8-10元杂环烯基、5-6元杂环烷基并5-6元杂环烯基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、5元或6元杂环烷基并5元或6元杂环烯基；所述杂环烷基、杂环烯基环上的-CH2-各自独立地任选被-C(＝O)-替换；所述杂芳基、杂环烷基、杂环烯基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环烷基、5元或6元杂环烯基、5元或6元杂环烷基并5元或6元杂环烯基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自5元杂芳基、苯基并5元或6元杂芳基、6元杂芳基并5元杂芳基、1,5-二氢-2H-吡咯-2-酮基、吡啶酮基、6元杂环烷基并吡啶酮基，所述杂芳基、杂环烷基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、6元杂芳基并5元杂芳基、1,5-二氢-2H-吡咯-2-酮基、吡啶酮基、6元杂环烷基并吡啶酮基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 所述异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自异噁唑基、吡唑基、异噻唑基、所述异噁唑基、吡唑基、异噻唑基、各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自所述 各自独立地被1个、2个、3个或4个Rd取代；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氨基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1个、2个或3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C1-4烷氧基、4-7元杂环烷基、苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、氰基或羟基；所述4-7元杂环烷基各自独立地含有1个O杂原子；所述苯基、噻吩基、噻唑基、 吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基是未取代的或被一个或多个选自F、Cl、氰基、甲基、乙基、丙基、环丙基、环丁基、环戊基、甲氧基、乙氧基、丙氧基、氟代甲基、氟代甲氧基的取代基取代；
    优选的，每个Rd各自独立地选自苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、氟取代的苯基、氯取代的苯基、氰基取代的苯基、甲基取代的苯基、甲氧基取代的苯基、三氟甲基取代的苯基、三氟甲氧基取代的苯基、氰基取代的吡啶基、氟取代的吡啶基、氯取代的吡啶基、甲基取代的吡啶基、甲氧基取代的吡啶基、甲基取代的吡唑基、三氟甲基取代的吡啶基、三氟甲氧基取代的吡啶基、氰基取代的吡啶基或羟基；
    优选的，每个Rd各自独立地选自苯基、噻吩基、吡啶基、羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl；
    或者优选的，每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、羟基或卤素，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5 环烷基的取代基取代；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1至3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3- 5环烷基的取代基取代；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C1-4卤代烷基、C5-6卤代烷基、C3-4环烷基、C5-6环烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、5元或6元杂芳基、苯基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；所述5元或6元杂芳基各自独立地含有1个、2个或3个选自N、O或S的杂原子；所述5元或6元杂芳基、苯基是未取代的或各自独立地被一个或多个选自卤素、氰基、C1-3烷基、C1-3卤代烷基、C1-3烷氧基、C1-3卤代烷氧基、C3-5环烷基的取代基取代；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C1-4卤代烷基、C3-4环烷基、C5-6环烷基、C1-4烷氧基、4-7元杂环烷基、苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1个O杂原子；所述苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基是未取代的或被一个或多个选自F、Cl、氰基、甲基、乙基、丙基、环丙基、环丁基、环戊基、甲氧基、乙氧基、丙氧基、氟代甲基、氟代甲氧基的取代基取代；
    优选的，每个Rd各自独立地选自苯基、噻吩基、噻唑基、吡唑基、吡咯基、呋喃基、噁唑基、吡啶基、嘧啶基、哒嗪基、吡嗪基、甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、氟取代的苯基、氯取代的苯基、氰基取代的苯基、甲基取代的苯基、甲氧基取代的苯基、三氟甲基取代的苯基、三氟甲氧基取代的苯基、氰基取代的吡啶基、氟取代的吡啶基、氯取代的吡啶基、甲基取代的吡啶基、甲氧基取代的吡啶基、甲基取代的吡唑基、三氟甲基取代的吡啶基、三氟甲氧基取代的吡啶基、氰基取代的吡啶基或羟基，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自苯基、噻吩基、吡啶基、羟基、甲基、异 丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自苯基、噻吩基、吡啶基、羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、 或Cl，其中有且只有一个Rd为羟基，且羟基取代在R2b与主基团连接位点的邻位；
    优选的，R2b选自
    最优选的，R2b选自
  6. 如权利要求1-3任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R2选自
    或者R2选自
    或者R2选自
    优选的，R2b选自5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基；所述杂芳基、杂环基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自5元杂芳基、苯基并5元或6元杂芳基、6元杂芳基并5元杂芳基、5元杂环基、6元杂环基、6元杂环基并吡啶酮基，所述杂芳基、杂环基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并6元杂芳基、6元杂芳基并5元杂芳基、5元杂环基、6元杂环基、6元杂环基并吡啶酮基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 所述异噁唑基、吡唑基、异噻唑基、苯并吡咯基、苯并吡啶基、吗啉 并吡啶酮基、1,5-二氢-2H-吡咯-2-酮基、 各自独立地被1个、2个、3个或4个Rd取代；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、C1-6烷氧基、4-7元杂环烷基、氨基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、氰基或羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C3-6环烷基、C1-4卤代烷基、C1-4烷氧基、4-7元杂环烷基、氰基或羟基；所述4-7元元杂环烷基各自独立地含有1个O杂原子；
    优选的，每个Rd各自独立地选自甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、 或羟基；
    优选的，每个Rd各自独立地选自羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、或Cl；
    或者优选的，每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、C1-6烷氧基、4-7元杂环烷基、羟基或卤素，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、氰基或羟基，其中至少一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、C1-6烷氧基、4-7元杂环烷基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1或2个选自N、O或S的杂原子；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C1-4卤代烷基、C5-6卤代烷基、C3-4环烷基、C5-6环烷基、C1-4烷氧基、C5-6烷氧基、4-7元杂环烷基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立 地含有1或2个选自N、O或S的杂原子；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C1-4卤代烷基、C3-6环烷基、C1-4烷氧基、4-7元杂环烷基、氰基或羟基，其中有且只有一个Rd为羟基；所述4-7元杂环烷基各自独立地含有1个O杂原子；
    优选的，每个Rd各自独立地选自甲基、乙基、丙基、异丙基、叔丁基、F、Cl、Br、环丙基、环丁基、环戊基、-CH2CF3、甲氧基、乙氧基、 或羟基，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、或Cl，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自羟基、甲基、异丙基、叔丁基、环丁基、环戊基、环丙基、-CH2CF3、甲氧基、或Cl，其中有且只有一个Rd为羟基，且羟基取代在R2b与主基团连接位点的邻位；
    优选的，R2b选自
    最优选的，R2b选自
  7. 如权利要求1-3任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R2选自
    或者R2选自
    或者R2选自优选的，R2b选自5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、6-10元芳基、5-10元杂环基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、苯基、奈基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基，所述杂芳基、杂环基各自独立地含有1-4个选自N、O或S的杂原子；所述5元杂芳基、8-10元杂芳基、5元或6元杂环基、8-10元杂环基各自独立地被一个或多个Rd取代；
    优选的，R2b选自5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基；所述杂芳基、杂环基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基、5元或6元杂环基、5元或6元杂环基并5元或6元杂环基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自5元杂芳基、苯基并6元杂芳基、6元杂芳基并5元杂芳基、6元杂环基、6元杂环基并吡啶酮基，所述杂芳基、杂环基各自独立地含有1-2个选自N、O或S的杂原子；所述5元杂芳基、苯基并6元杂芳基、6元杂芳基并5元杂芳基、6元杂环基、6元杂环基并吡啶酮基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自吡唑基、噻唑基、噻吩基、异噻唑基、苯基并吡啶基、吡啶并吡咯基、吗啉基并吡啶酮基、吡啶酮基；所述吡唑基、噻唑基、噻吩基、异噻唑基、苯基并吡啶基、吡啶并吡咯基、吗啉基并吡啶酮基、吡啶酮基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基或羟基；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C3-4环烷基、C5-6环烷基、C1-4卤代烷基、C5-6卤代烷基、氰基或羟基；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C3-6环烷基、C1-4卤代烷基、氰基或羟基；
    优选的，每个Rd各自独立地选自甲基、乙基、丙基、F、Cl、Br、环丙基、环丁基、环戊基、三氟甲基或羟基；
    优选的，每个Rd各自独立地选自甲基、Cl、Br、环丙基或羟基；
    或者优选的，每个Rd各自独立地选自C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基、氨基、羟基或卤素，其中至少一个Rd为羟基；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基或羟基，其中至少一个Rd为羟基；
    优选的，每个Rd各自独立地选自卤素、C1-6烷基、C3-6环烷基、C1-6卤代烷基、氰基或羟基，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C5-6烷基、C1-4卤代烷基、C5-6卤代烷基、C3-4环烷基、C5-6环烷基、氰基或羟基，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自卤素、C1-4烷基、C1-4卤代烷基、C3-6环烷基、氰基或羟基，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自甲基、乙基、丙基、F、Cl、Br、环丙基、环丁基、环戊基、三氟甲基或羟基，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自甲基、Cl、Br、环丙基或羟基，其中有且只有一个Rd为羟基；
    优选的，每个Rd各自独立地选自甲基、Cl、Br、环丙基或羟基，其中有且只有一个Rd为羟基，且羟基取代在R2b与主基团连接位点的邻位；
    优选的，R2b选自
    各自独立地可进一步被1个、2个或3个Rd取代；
    最优选的，R2b选自
    或者优选的，R2b选自5元杂芳基，所述5元杂芳基各自独立地被1个、2个、3个或4个Rd取代；
    进一步优选的，R2b选自吡唑基、噻唑基、噻吩基、异噻唑基，所述吡唑基、噻唑基、噻吩基、异噻唑基各自独立地被1个、2个、3个或4个Rd取代；
    进一步优选的，R2b选自 可进一步被1个、2个或3个Rd取代；
    进一步优选的，R2b选自可进一步被1个、2个或3个Rd取代；
    或者优选的，R2b选自：
    进一步优选的，R2b选自：
    优选的，R2b选自6元杂环基、6元杂环基并6元杂环基，所述杂环基含有1-2个选自N、O或S的杂原子；所述6元杂环基、6元杂环基并6元杂环基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自吗啉基并吡啶酮基、吡啶酮基；所述吗啉基并吡啶酮基、吡啶酮基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自吡啶酮基；所述吡啶酮基被1个、2个、3个或4个Rd取代；
    优选的，R2b选自2-吡啶酮基；所述2-吡啶酮基被1个、2个、3个或4个Rd取代；
    优选的，R2b选自可进一步被1个、2个或3个Rd取代；
    优选的，R2b选自可进一步被1个、2个或3个Rd取代；
    优选的，R2b选自
    或者优选的，R2b选自
    或者优选的，R2b选自苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基；所述杂芳基含有1-2个选自N、O或S的杂原子；所述苯基并5元或6元杂芳基、5元或6元杂芳基并5元或6元杂芳基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自苯基并6元杂芳基、6元杂芳基并5元杂芳基，所述杂芳基含有1-2个选自N、O或S的杂原子；所述苯基并6元杂芳基、6元杂芳基并5元杂芳基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自苯基并吡啶基、吡啶并吡咯基；所述苯基并吡啶基、吡啶并吡咯基各自独立地被1个、2个、3个或4个Rd取代；
    优选的，R2b选自可进一步被1个、2个或3个Rd取代；
    优选的，R2b选自
  8. 如权利要求1-7任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R1选自卤素、C1-6烷基、5元或6元环烯基、5-10元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所述烷基、环烯基、杂环基、杂芳基、苯基各自独立地是未取代的或被1个或多个Rg取代；
    优选的，R1选自5元或6元环烯基、5-10元杂环基、5元或6元杂芳基、苯基，所述杂环基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所述环烯基、杂环基、杂芳基、苯基各自独立地是未取代的或被1个或多个Rg取代；
    优选的，R1选自5元或6元环烯基、5-10元杂环烷基、5元或6元杂环烯基、5元或6元杂芳基、苯基，所述杂环烷基、杂环烯基、杂芳基各自独立地含有1-2个选自N、O或S的杂原子，所述环烯基、杂环烷基、杂环烯基、杂芳基、苯基各自独立地是未取代的或被1个或多个Rg取代；
    优选的，R1选自6元环烯基、6-9元杂环烷基、6元杂环烯基、吡啶基、嘧啶基、哒嗪基，所述杂环烷基、杂环烯基各自独立地含有1-2个选自N、O或S的杂原子，所述环烯基、杂环烷基、杂环烯基各自独立地是未取代的或被1个或多个Rg取代；
    优选的，每个Rg各自独立地选自羟基、卤素、C1-4烷基、C1-4烷氧基、＝O、R’R”N-；所述烷基、烷氧基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；其中，R’、R”各自独立地选自H、C1-4烷基、C1- 4卤代烷基；
    优选的，每个Rg各自独立地选自羟基、卤素、C1-2烷基、C3-4烷基、C1-2烷氧基、C3-4烷氧基、＝O、R’R”N-；所述烷基、烷氧基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；其中，R’、R”各自独立地选自H、C1-2烷基、C3-4烷基、C1-2卤代烷基、C3-4卤代烷基；
    优选的，每个Rg各自独立地选自羟基、F、Cl、C1-2烷基、C1-2烷氧基、＝O、R’R”N-；所述烷基、烷氧基各自独立地是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；其中，R’、R”各自独立地选自H、C1-2烷基、C1- 2卤代烷基；
    优选的，每个Rg各自独立地选自羟基、F、甲基、甲氧基、＝O、R’R”N-；所述甲基是未取代的或被1或多个选自F、Cl、羟基、甲氧基的取代基取代；其中，R’、R”各自独立地选自H、甲基；
    优选的，R1选自
    优选的，R1选自
    最优选的，R1选自
  9. 如权利要求1-7任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R1选自卤素、C1-6烷基、5元或6元杂环基，所述杂环基含有1-2个选自N、O或S的杂原子，所述烷基、杂环基是未取代的或被1个或多个Rg取代；
    优选的，R1选自5元或6元杂环基，所述杂环基含有1-2个选自N、O或S的杂原子，所述杂环基是未取代的或被1个或多个Rg取代；
    优选的，R1选自5元或6元杂环烷基、5元或6元杂环烯基，所述杂环烷基、杂环烯基含有1-2个选自N、O或S的杂原子，所述杂环烷基、杂环烯基是未取代的或被1个或多个Rg取代；
    优选的，R1选自5元或6元杂环烯基，所述杂环烯基含有1-2个选自N或O的杂原子，所述杂环烯基是未取代的或被1个或多个Rg取代；
    或者优选的，R1选自 是未取代的或被1个或多个Rg取代；
    优选的，每个Rg各自独立地选自羟基、卤素、C1-4烷基、C1-4烷氧基；所述C1-4烷基、C1-4烷氧基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
    优选的，每个Rg各自独立地选自羟基、卤素、C1-2烷基、C3-4烷基、C1-2烷氧基、C3-4烷氧基；所述烷基、烷氧基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
    优选的，每个Rg各自独立地选自羟基、F、Cl、C1-2烷基、C1-2烷氧基；所 述烷基、烷氧基是未取代的或被1或多个选自卤素、羟基、C1-2烷氧基的取代基取代；
    优选的，每个Rg各自独立地选自羟基、F、甲基、乙基；所述甲基、乙基是未取代的或被1或多个选自卤素、羟基、甲氧基的取代基取代；
    优选的，R1选自
    优选的，R1选自
    优选的，R1选自
  10. 如权利要求1-9任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R3选自C1-4烷基，所述烷基是未取代的或被1个、2个或3个选自羟基或卤素的取代基取代；
    优选的，R3选自C1-4烷基，所述烷基是未取代的；
    优选的，R3选自C1-2烷基，所述烷基是未取代的；
    优选的，R3选自甲基或乙基；
    优选的，R3选自乙基。
  11. 如权利要求1-10任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：
    R4选自H、卤素、C1-4烷基，所述烷基是未取代的或被1个、2个或3个卤素取代；
    优选的，R4选自H、卤素、C1-4烷基，所述烷基是未取代的或被1个、2个或3个F取代；
    优选的，R4选自H、F、Cl、Br、C1-4烷基，所述C1-4烷基是未取代的；
    优选的，R4选自H、F、Cl、Br、C1-2烷基，所述C1-2烷基是未取代的或被1个、2个或3个F取代；
    优选的，R4选自H、F、Cl、Br、甲基、乙基，所述甲基、乙基各自独立地是未取代的或被1个、2个或3个F取代；
    优选的，R4选自H、F、Cl、Br、甲基、-CF3；
    优选的，R4选自Cl、甲基、-CF3；
    优选的，R4选自H、F、Cl、Br、甲基、乙基，所述甲基、乙基是未取代的；
    优选的，R4选自H、F、Cl、Br或甲基；
    优选的，R4选自Cl或甲基；
    或者，
    R5选自H、甲基、卤素；
    优选的，R5选自H、F、Cl或甲基；
    优选的，R5选自H、Cl或甲基；
    优选的，R5选自H；
    或者，
    R6选自H、卤素、C1-4烷基；所述烷基是未取代的或被1个、2个或3个卤素取代；
    优选的，R6选自H、卤素、C1-4烷基，所述烷基是未取代的或被1个、2个或3个F取代；
    优选的，R6选自H、卤素、C1-2烷基，所述烷基是未取代的或被1个、2个或3个F取代；
    优选的，R6选自H、F、Cl、甲基、乙基，所述甲基、乙基是未取代的或被1个、2个或3个F取代；
    优选的，R6选自H、F、Cl、甲基或-CF3；
    优选的，R6选自H、F或Cl；
    或者优选的，R6选自H、卤素、C1-4烷基，所述烷基被1个、2个或3个F取代；
    优选的，R6选自H、卤素、C1-2烷基，所述烷基被1个、2个或3个F取代；
    优选的，R6选自H、F、Cl、甲基、乙基，所述甲基、乙基被1个、2个或3个F取代；
    优选的，R6选自H、F、Cl或-CF3；
    优选的，R6选自H、F或Cl；
    或者，
    R7选自H、-SF5、-C(O)H、卤素、C1-4烷基、C2-4烯基、C2-4炔基、C3-5环烷基；所述烷基、环烷基是未取代的或被1个或多个卤素取代；
    优选的，R7选自-SF5、-C(O)H、卤素、C1-4烷基；所述烷基是未取代的或被1个、2个或3个卤素取代；
    优选的，R7选自-SF5、-C(O)H、卤素、C1-4烷基；所述烷基是未取代的或被1个、2个或3个F取代；
    优选的，R7选自-SF5、-C(O)H、卤素、C1-2烷基；所述烷基是未取代的或被1个、2个或3个F取代；
    优选的，R7选自-SF5、-C(O)H、F、Cl、Br、甲基、乙基；所述甲基、乙基 是未取代的或被1个、2个或3个F取代；
    优选的，R7选自-CF3、-CHF2、-CH2CH3、Cl、Br、-SF5或-C(O)H；
    优选的，R7选自-CF3；
    优选的，R7选自-OCF3；
    或者，
    X选自N或CR8，R8选自H、F、Cl或Br；
    优选的，R8选自H或F；
    优选的，功能基团选自
    优选的，功能基团选自
    优选的，功能基团选自
    优选的，功能基团选自
    优选的，功能基团选自
  12. 如权利要求1-11任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：式(I)具有选自式(IB)所示通式结构：
    其中，R1、R2b、R3、R4各自定义如式(I)中所述定义；
    优选的，式(I)具有式(IB-1)所示结构：
    其中，R1、R2b、R4定义如式(I)中所述定义。
  13. 如权利要求1-12任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：式(I)具有选自式(IIA)、式(IIB)所示结构：
    其中，R1、R2b、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIA)具有式(IIA-1)或式(IIA-2)或式(IIA-3)所示结构：
    其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIA-1)具有式(IIA-1-1)所示结构：
    其中，R1、Rd定义如式(I)中所述定义；
    其中，R1、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIA-2)具有式(IIA-2-1)所示结构：
    其中，R1定义如式(I)中所述定义；
    其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIA-3)具有式(IIA-3-1)所示结构：
    其中，R1、Rd定义如式(I)中所述定义；
    其中，R1、R2b、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIB)具有式(IIB-1)或式(IIB-2)或式(IIB-3)所示结构：
    其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIB-1)具有式(IIB-1-1)所示结构：
    其中，R1、Rd定义如式(I)中所述定义；
    其中，R1、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIB-2)具有式(IIB-2-1)所示结构：
    其中，R1定义如式(I)中所述定义；
    其中，R1、Rd、R4、R5、R6、R7定义如式(I)中所述定义；
    进一步的，所述式(IIB-3)具有式(IIB-3-1)所示结构：
    其中，R1、Rd定义如式(I)中所述定义。
  14. 如权利要求1-12任一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：式(I)具有选自式(IIA-4)或式(IIB-4)所示结构：
    其中，环B为5元杂芳基；所述5元杂芳基含有1-2个选自N、O和S的杂原子；所述5元杂芳基为未取代的或被1个或多个C1-4烷基取代；
    优选的，环B为5元杂芳基；所述5元杂芳基含有1个选自N、O和S的杂原子；所述5元杂芳基为未取代的或被1个或多个C1-4烷基取代；
    优选的，环B选自呋喃基、吡咯基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基；所述呋喃基、吡咯基、噻吩基、吡唑基、咪唑基、噻唑基、异噻唑基、噁唑基、异噁唑基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；
    优选的，环B选自呋喃基、吡咯基、噻吩基；所述呋喃基、吡咯基、噻吩基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；
    优选的，环B选自呋喃基、吡咯基；所述呋喃基、吡咯基各自独立地为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；
    优选的，环B选自呋喃基、吡咯基、噻吩基；所述吡咯基为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；
    优选的，环B选自呋喃基、吡咯基；所述吡咯基为未取代的或被1个或多个选自甲基、乙基、正丙基、异丙基、正丁基、仲丁基、异丁基、叔丁基的取代基取代；
    优选的，环B选自呋喃基、吡咯基、噻吩基；所述吡咯基为未取代的或被1个甲基取代；
    优选的，环B选自呋喃基、吡咯基；所述吡咯基为未取代的或被1个甲基取代。
  15. 如权利要求1所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，其特征在于：式(I)选自下述具体化合物：









  16. 一种治疗和/或预防与WRN生物学活性相关的疾病的药物组合物，其特征在于：所述药物组合物包括治疗和/或预防有效量的如权利要求1-15任意一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，以及任选的药用辅料；
    优选的，所述与WRN生物学活性相关的疾病为肿瘤或癌症；
    优选的，所述与WRN生物学活性相关的疾病是MSI肿瘤或MSI癌症；
    优选的，一种药物组合物，其特征在于：包含如权利要求1-15任意一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，以及任选的药用辅料。
  17. 如权利要求1-15任意一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如权利要求16所述的药物组合物，在制备治疗和/或预防与WRN生物学活性相关的疾病的药物中，和/或，在治疗和/或预防与WRN生物学活性相关的疾病中的应用；
    优选的，所述与WRN生物学活性相关的疾病为肿瘤或癌症；
    优选的，所述与WRN生物学活性相关的疾病是MSI肿瘤或MSI癌症。
  18. 一种治疗和/或预防与WRN生物学活性相关的疾病的方法，其包括给予有需要的个体治疗和/或预防有效量的如权利要求1-15任意一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如权利要求16所述的药物组合物；
    优选的，所述与WRN生物学活性相关的疾病为肿瘤或癌症；
    优选的，所述与WRN生物学活性相关的疾病是MSI肿瘤或MSI癌症。
  19. 如权利要求1-15任意一项所述的化合物，或其立体异构体、互变异构体、多晶型物、共晶物、溶剂化物、代谢物、前药、氘代化合物、药学上可接受的盐，或者如权利要求16所述的药物组合物，其用于治疗和/或预防与WRN生物学活性相关的疾病，
    优选的，所述与WRN生物学活性相关的疾病为肿瘤或癌症；
    优选的，所述与WRN生物学活性相关的疾病是MSI肿瘤或MSI癌症。
PCT/CN2024/072876 2023-01-18 2024-01-17 双杂环wrn抑制剂、其制备方法及其应用 WO2024153155A1 (zh)

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