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WO2020187291A1 - 吡唑并嘧啶化合物和药物组合物及其应用 - Google Patents

吡唑并嘧啶化合物和药物组合物及其应用 Download PDF

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WO2020187291A1
WO2020187291A1 PCT/CN2020/080198 CN2020080198W WO2020187291A1 WO 2020187291 A1 WO2020187291 A1 WO 2020187291A1 CN 2020080198 W CN2020080198 W CN 2020080198W WO 2020187291 A1 WO2020187291 A1 WO 2020187291A1
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alkyl
substituted
halogen
group
alkyl group
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PCT/CN2020/080198
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English (en)
French (fr)
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杨光富
黄伟
卓林胜
徐红闯
王明书
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华中师范大学
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Priority to EP20774324.6A priority Critical patent/EP3932923B1/en
Priority to AU2020242735A priority patent/AU2020242735B2/en
Priority to US17/440,362 priority patent/US20220177477A1/en
Priority to JP2022504321A priority patent/JP7209415B2/ja
Publication of WO2020187291A1 publication Critical patent/WO2020187291A1/zh

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to the field of biomedicine, in particular to a pyrazolopyrimidine compound, a pharmaceutical composition containing the pyrazolopyrimidine compound, the aforementioned pyrazolopyrimidine compound and the application of the pharmaceutical composition.
  • NTRK/TRK Tropomyosin receptor kinase
  • TRK family mainly includes 3 members, NTRK1/TRKA, NTRK2/TRKB and NTRK3/TRKC.
  • the complete TRK kinase includes three parts: extracellular domain, transmembrane domain and intracellular domain. After the extracellular domain of TRK kinase is combined with the corresponding ligand, it can cause the kinase configuration to change and form a dimer.
  • TRK kinase undergoes autophosphorylation to activate its own kinase activity, which further activates downstream signal transduction pathways (such as MAPK, AKT, PKC, etc.) to produce corresponding biological functions; among them, NGF (Nerve Growth Factor)
  • BDNF derived neurotrophic factor
  • NT3 neurotrophic factor 3
  • TRK kinases play important physiological functions in the development of nerves, including the growth and function maintenance of neuronal axons, the occurrence and development of memory, and the protection of neurons from injury. At the same time, a large number of studies have shown that the activation of the TRK signal transduction pathway is closely related to the occurrence and development of tumors. Activated TRK signal proteins have been found in neurocytoma, prostate cancer, and breast cancer.
  • TRK fusion proteins have shown their biological function of promoting tumorigenesis.
  • the earliest TPM3-TRKA fusion protein was found in colon cancer cells, with an incidence of about 1.5% in clinical patients tested.
  • TRK fusion proteins such as CD74-NTRK1, MPRIP-NTRK1, QKI-NTRK2, ETV6, were found in different types of clinical tumor patient samples such as lung cancer, head and neck cancer, breast cancer, thyroid cancer, glioma, etc. -NTRK3, BTB1-NTRK3, etc.
  • These different NTRK fusion proteins are in a highly activated kinase activity state without ligand binding, so they can continuously phosphorylate downstream signal pathways, induce cell proliferation, and promote tumor occurrence and development.
  • TRK fusion protein has become an effective anti-cancer target and research hotspot.
  • WO2010048314, WO2012116217, WO2011146336, WO2010033941, WO2018077246, etc. all disclose TRK kinase inhibitors with different structural types.
  • TRK mutations that appear after continuous administration are an important reason for tumor drug resistance.
  • TRKA G595R, G667C, G667S and F589L mutations (Russo M, etc. Cancer Discovery, 2016, 6(1), 36-44), TRKC G623R and G696A mutations (Drilon A. et al. Annals of Oncology 2016, 27(5), 920-926), and looking for new TRK kinase inhibitors is expected to solve TRK mutations Tumor drug resistance caused by problems.
  • nitrogen-containing aromatic heterocycles usually have good druggability.
  • a typical example is the ALK kinase inhibitor crizotinib (Cui J. et al. J. Med. Chem. 2011, 54, 6342-6363).
  • WO2007147647 and WO2007025540 also respectively disclose pyrazole-substituted pyrazolopyridine compounds and pyrazole-substituted imidazopyridazine compounds as ALK kinase inhibitors and their applications in disease treatment.
  • One of the objectives of the present invention is to provide a new pyrazolopyrimidine compound with excellent antitumor activity.
  • the first aspect of the present invention provides a pyrazolopyrimidine compound having a structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, or mutual Tautomers, nitrogen oxides, hydrates, solvates, metabolites, or prodrugs thereof,
  • R 1, R 2, R 3 and R 4 are each independently selected from H, halo, C 1-12 alkyl group, a halogen-substituted alkyl having 1 to 6 of C 1-12;
  • R 5 is selected from H, C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl, hydroxy substituted C 1-12 alkyl, C 2-12 alkoxy-substituted alkyl, cyano-substituted C 2-12 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-12 cycloalkyl;
  • R 6 is selected from H, C 1-12 alkyl, hydroxy substituted C 1-12 alkyl and halogen;
  • R 7 is selected from H, C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl, hydroxy substituted C 1-12 alkyl group, a cyano-substituted C 2-12 Alkyl groups, C 2-12 cycloalkyl groups containing 1-3 heteroatoms selected from N, O and S, C 2-12 acyl groups, and sulfonyl groups.
  • the second aspect of the present invention provides the pyrazolopyrimidine compound having the structure represented by formula (I) or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, Application of tautomers, nitrogen oxides, hydrates, solvates, metabolites, or prodrugs thereof in the preparation of drugs for preventing and/or treating TRK kinase-mediated diseases.
  • the third aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or diluent, and as an active ingredient the structure represented by the first aspect of the present invention having formula (I)
  • the pyrazolo pyrimidine compound or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, or prodrug thereof having formula (I)
  • the pyrazolo pyrimidine compound or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, or prodrug thereof The pyrazolo pyrimidine compound or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate, solvate, metabolite, or prodrug thereof .
  • the fourth aspect of the present invention provides the use of the pharmaceutical composition described in the third aspect of the present invention in the preparation of a medicine for preventing and/or treating TRK kinase receptor-mediated diseases.
  • the fifth aspect of the present invention provides the pyrazolo pyrimidine compound having the structure represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer or geometric isomer thereof as described in the first aspect of the present invention , Tautomers, nitrogen oxides, hydrates, solvates, metabolites, or prodrugs thereof, or the composition of the third aspect of the present invention is used in the preparation of drugs for the prevention and/or treatment of tumors In the application.
  • the pyrazolopyrimidine compound having the structure represented by formula (I) or its pharmaceutically acceptable salt, or its stereoisomer, geometric isomer, tautomer, nitrogen oxide, hydrate Compounds, solvates, metabolites, or their prodrugs show excellent inhibitory activity on TRK kinase, and at the same time, show good anti-tumor activity at the animal level.
  • the first aspect of the present invention provides a pyrazolopyrimidine compound or a pharmaceutically acceptable salt thereof, or a stereoisomer, geometric isomer, tautomer, nitrogen oxide, Hydrates, solvates, metabolites, or prodrugs thereof.
  • C 1-12 alkyl group means an alkyl group with a total number of carbon atoms of 1-12, including straight-chain alkyl, branched-chain alkyl or cycloalkyl, for example, the total number of carbon atoms is 1, 2, 3, or 4.
  • C 1-12 alkyl group substituted by 1-6 halogens means an alkyl group with a total number of carbon atoms of 1-12, including straight chain alkyl, branched chain alkyl and cycloalkyl, and the C 1-12 The 1-6 H in the alkyl group is substituted by a halogen atom selected from halogen.
  • 1, 2, 3, 4, 5 or 6 H in the C 1-12 alkyl group are selected from fluorine, chlorine, Any one or more halogen atoms of bromine and iodine may be substituted by, for example, trifluoromethyl, difluoromethyl, monofluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, etc.
  • halogen atoms of bromine and iodine may be substituted by, for example, trifluoromethyl, difluoromethyl, monofluoromethyl, monofluoroethyl, difluoroethyl, trifluoroethyl, etc.
  • C 1-8 alkyl substituted by 1-6 halogens and "C 1-6 alkyl substituted by 1-6 halogens" except that the carbon atom The numbers are different.
  • Haldroxy-substituted C 1-12 alkyl group means an alkyl group with a total number of carbon atoms of 1-12, including straight chain alkyl, branched chain alkyl or cycloalkyl, and the C 1-12 alkyl group At least one H is substituted by a hydroxyl group.
  • Alkoxy-substituted C 2-12 alkyl group means a group with a total number of carbon atoms of 2-12, and the structural formula of this group can be expressed as -R 1 OR 2 , where in R 1 and R 2 The total number of carbon atoms is 2-12, and R 1 is directly connected to the phenoxy group in the pyrazolopyrimidine compound of the structure represented by formula (I) of the present invention.
  • the "cyano-substituted C 2-12 alkyl group” means an alkyl group having a total number of 2-12 carbon atoms, including straight chain alkyl, branched chain alkyl or cycloalkyl, and the C 2-12 alkyl group At least one H of is substituted by a cyano group, and the carbon atoms in the "cyano group” are counted in the total number of carbon atoms in the group.
  • C 2-12 cycloalkyl containing 1-3 heteroatoms selected from N, O, and S means a cycloalkyl group having a total of 2-12 carbon atoms and 1-3 of the atoms forming the ring
  • the atom is selected from heteroatoms of N, O and S, and the atoms forming the ring may contain an alkyl substituent, and the number of carbon atoms contained in the alkyl substituent is included in the aforementioned range of the total number of carbon atoms.
  • the "C 2-12 cycloalkyl group containing 1-3 heteroatoms selected from N, O, and S" can be, for example, a three-membered ring, a four-membered ring, a five-membered ring, a six-membered ring, a seven-membered ring, and an eight-membered ring.
  • -Membered ring, nine-membered ring, ten-membered ring, eleven-membered ring or twelve-membered ring, and the H in the cycloalkyl group may be optionally substituted or unsubstituted with substituents. If substituted, the substituents are independent Ground is selected from at least one of halogen, hydroxyl, nitro and mercapto.
  • C 2-12 acyl group means an acyl group with a total number of carbon atoms of 2-12, and may be, for example, acetyl, propionyl and the like.
  • the "sulfonyl group” may contain a C 1-6 alkyl group, and the sulfonyl group may be represented by, for example, -SO 2 R 3 , where R 3 may be a C 1-6 alkyl group.
  • R 1 , R 2 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, bromine, C 1-8 Alkyl, C 1-8 alkyl substituted by 1-6 halogens selected from fluorine, chlorine and bromine; more preferably, R 1 , R 2 , R 3 and R 4 are each independently selected from H, fluorine , chloro, bromo, C 1-6 alkyl, a 1-4 heteroatoms selected from fluorine, chlorine and bromine halogen-substituted C 1-6 alkyl; and more preferably, R 1, R 3 and R 4 are each Independently selected from H, fluorine, chlorine, bromine, C 1-8 alkyl, C 1-8 alkyl substituted with 1-6 halogens selected from fluorine, chlorine and bromine; and R 2 is H or F.
  • R 1 , R 3 and R 4 are each independently selected from H, halogen, C 1- 12 alkyl groups, C 1-12 alkyl groups substituted by 1-6 halogens; and R 2 is halogen; more preferably, R 1 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, Bromine, C 1-8 alkyl, C 1-8 alkyl substituted with 1-6 halogens selected from fluorine, chlorine and bromine; and R 2 is F; more preferably, R 1 , R 3 and R 4 is H; R 2 is F.
  • R 5 is selected from H, C 1-8 alkyl, and substituted with 1-6 halogens selected from fluorine, chlorine and bromine.
  • R 6 is preferably selected from H, C 1-12 alkyl, hydroxy substituted C 1-12 alkyl and halogen; more preferably Preferably, R 6 is selected from H, C 1-8 alkyl, hydroxy-substituted C 1-8 alkyl and halogen; further preferably, R 6 is selected from H, C 1-6 alkyl, hydroxy-substituted C 1-6 alkyl and halogen.
  • R 7 is selected from H, C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl group , hydroxy-substituted C 1-12 alkyl group, a cyano-substituted C 2-12 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-12 cycloalkyl, C 2-12 acyl, sulfonyl; more preferably, R 7 is selected from H, C 1-8 alkyl, and C 1-8 alkyl substituted with 1-6 halogens selected from fluorine, chlorine and bromine , Hydroxy substituted C 1-8 alkyl, cyano substituted C 2-8 alkyl, C 2-10 cycloalkyl containing 1-3 heteroatoms selected from N, O and S, C 2-8 acyl, sulfonyl; more preferably, R 7 is selected from H, C 1-12 alkyl group, a halogen-substituted 1-6
  • the inventors of the present invention found that when the R 2 is halogen, the pyrazolo pyrimidine compound of the formula (I) provided by the present invention has a better effect on TRK kinase, especially mutant TRK kinase. High inhibitory activity; at the same time, it exhibits more reasonable pharmacokinetic properties and better anti-tumor activity in vivo.
  • R 1 , R 2 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, bromine, C 1-8 alkyl, C 1 substituted by 1-6 halogens selected from fluorine, chlorine and bromine -8 alkyl;
  • R 5 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, an alkoxy substituted C 2-8 alkyl group, a cyano group-substituted C 2-8 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-10 cycloalkyl;
  • R 6 is selected from H, C 1-8 alkyl, hydroxy substituted C 1-8 alkyl and halogen;
  • R 7 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, a cyano Group substituted C 2-8 alkyl, C 2-10 cycloalkyl containing 1-3 heteroatoms selected from N, O and S, C 2-8 acyl, sulfonyl.
  • R 1 , R 2 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, bromine, C 1-6 alkyl, C 1 substituted by 1-4 halogens selected from fluorine, chlorine and bromine -6 alkyl;
  • R 5 is selected from H, C 1-6 alkyl, a 1-4 heteroatoms selected from fluorine, chlorine and bromine halogen substituted C 1-6 alkyl, hydroxy substituted C 1-6 alkyl group, an alkoxy An oxy-substituted C 2-8 alkyl group, a cyano group-substituted C 2-6 alkyl group, a C 2-8 cycloalkyl group containing 1-3 heteroatoms selected from N, O and S;
  • R 6 is selected from H, C 1-6 alkyl, hydroxy substituted C 1-6 alkyl and halogen;
  • R 7 is selected from H, C 1-6 alkyl, a 1-4 heteroatoms selected from fluorine, chlorine and bromine halogen-substituted C 1-6 alkyl group, a hydroxyl group substituted with a C 1-6 alkyl group, a cyano substituted C 2-6 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-8 cycloalkyl, C 2-6 acyl, sulfonyl group.
  • R 1 , R 2 , R 3 , R 4 and R 5 contains an F atom; and R 5 is selected A C 1-12 alkyl group substituted with 1-6 halogens; further preferably, at least one of R 1 , R 2 , R 3 , R 4 and R 5 contains an F atom; and R 5 is selected from 1-6 C 1-8 alkyl groups substituted with halogens selected from fluorine, chlorine and bromine.
  • R 1, R 2, R 3 and R 4 are each independently selected from H, halo, C 1-12 alkyl group, a halogen-substituted alkyl having 1 to 6 of C 1-12;
  • R 5 is -CH 2 CHF 2 ;
  • R 6 is selected from C 1-12 alkyl, hydroxy-substituted C 1-12 alkyl and halogen;
  • R 7 is selected from H, C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl, hydroxy substituted C 1-12 alkyl group, a cyano-substituted C 2-12 Alkyl groups, C 2-12 cycloalkyl groups containing 1-3 heteroatoms selected from N, O and S, C 2-12 acyl groups, and sulfonyl groups.
  • R 1 , R 2 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, bromine, C 1-8 alkyl, C 1 substituted by 1-6 halogens selected from fluorine, chlorine and bromine -8 alkyl;
  • R 5 is -CH 2 CHF 2 ;
  • R 6 is selected from C 1-8 alkyl, hydroxy substituted C 1-8 alkyl and halogen;
  • R 7 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, a cyano Group substituted C 1-8 alkyl, C 2-10 cycloalkyl containing 1-3 heteroatoms selected from N, O and S, C 2-8 acyl, sulfonyl.
  • R 1 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, bromine, C 1-8 alkyl, and C 1-8 substituted by 1-6 halogens selected from fluorine, chlorine and bromine alkyl;
  • R 2 is H or F
  • R 5 is -CH 2 CHF 2 ;
  • R 6 is selected from C 1-8 alkyl, hydroxy substituted C 1-8 alkyl and halogen;
  • R 7 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, a cyano Group substituted C 2-8 alkyl, C 2-10 cycloalkyl containing 1-3 heteroatoms selected from N, O and S, C 2-8 acyl, sulfonyl.
  • the compound of the structure represented by formula (I) is selected from at least one of the following compounds:
  • R 7 is selected from C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl group, a hydroxyl group substituted C 1-12 alkyl group, a cyano group-substituted C 2-12 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-12 cycloalkyl, C 2- an acyl group, a sulfonyl group 12; more preferably, R 7 is selected from C 1-6 alkyl, substituted with 1-4 substituents selected from fluorine, chlorine and bromine halogen substituted C 1-6 alkyl group, a hydroxyl group alkyl, cyano substituted C 1-6 alkyl group a C 2-6 containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-8 cycloalkyl, C 2-6 of Acyl, sulfony
  • R 1, R 3 and R 4 are each independently selected from H, halo, C 1-12 alkyl group, a halogen-substituted alkyl having 1 to 6 of C 1-12;
  • R 2 is halogen
  • R 5 is selected from H, C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl, hydroxy substituted C 1-12 alkyl, C 2-12 alkoxy-substituted alkyl, cyano-substituted C 2-12 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-12 cycloalkyl;
  • R 6 is selected from C 1-12 alkyl, hydroxy-substituted C 1-12 alkyl and halogen;
  • R 7 is selected from C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl, hydroxy substituted C 1-12 alkyl group, a cyano-substituted C 2-12 alkyl group , C 2-12 cycloalkyl, C 2-12 acyl, sulfonyl containing 1-3 heteroatoms selected from N, O and S.
  • R 1 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, bromine, C 1-8 alkyl, and C 1-8 substituted by 1-6 halogens selected from fluorine, chlorine and bromine alkyl;
  • R 2 is F
  • R 5 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, an alkoxy substituted C 2-8 alkyl group, a cyano group-substituted C 2-8 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-10 cycloalkyl;
  • R 6 is selected from C 1-8 alkyl, hydroxy substituted C 1-8 alkyl and halogen;
  • R 7 is selected from C 1-8 alkyl by 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl, cyano-substituted C 1-8 alkyl, C 2-10 cycloalkyl containing 1-3 heteroatoms selected from N, O and S, C 2-8 acyl, sulfonyl.
  • R 1 , R 3 and R 4 are all H; R 2 is F;
  • R 5 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, an alkoxy substituted C 2-8 alkyl group, a cyano group-substituted C 2-8 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-10 cycloalkyl;
  • R 6 is selected from C 1-8 alkyl, hydroxy substituted C 1-8 alkyl and halogen;
  • R 7 is selected from C 1-8 alkyl by 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl, cyano-substituted C 2-8 alkyl group, C 2-10 cycloalkyl group containing 1-3 heteroatoms selected from N, O and S, C 2-8 acyl group, sulfonyl group.
  • the compound of the structure represented by formula (I) is selected from at least one of the following compounds:
  • R 7 is H.
  • R 1, R 3 and R 4 are each independently selected from H, halo, C 1-12 alkyl group, a halogen-substituted alkyl having 1 to 6 of C 1-12;
  • R 2 is halogen
  • R 5 is selected from H, C 1-12 alkyl group, a halogen-substituted 1-6 C 1-12 alkyl, hydroxy substituted C 1-12 alkyl, C 2-12 alkoxy-substituted alkyl, cyano-substituted C 2-12 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-12 cycloalkyl;
  • R 6 is selected from a C 1-12 alkyl group, a hydroxy-substituted C 1-12 alkyl group and halogen.
  • R 1 , R 3 and R 4 are each independently selected from H, fluorine, chlorine, bromine, C 1-8 alkyl, and C 1-8 substituted by 1-6 halogens selected from fluorine, chlorine and bromine alkyl;
  • R 2 is F
  • R 5 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, an alkoxy substituted C 2-8 alkyl group, a cyano group-substituted C 2-8 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-10 cycloalkyl;
  • R 6 is selected from a C 1-8 alkyl group, a hydroxy-substituted C 1-8 alkyl group and halogen.
  • R 1 , R 3 and R 4 are all H; R 2 is F;
  • R 5 is selected from H, C 1-8 alkyl, a 1-6 groups selected from fluorine, chlorine and bromine halogen-substituted C 1-8 alkyl, hydroxy substituted C 1-8 alkyl group, an alkoxy substituted C 2-8 alkyl group, a cyano group-substituted C 2-8 alkyl group containing 1-3 heteroatoms selected from N, C, O and S heteroatoms 2-10 cycloalkyl;
  • R 6 is selected from a C 1-8 alkyl group, a hydroxy-substituted C 1-8 alkyl group and halogen.
  • the compound of the structure represented by formula (I) is selected from at least one of the following compounds:
  • Compound b1-1 Compound b2-5: Compound b3-6: Compound b4-7: Compound b5-11: Compound b6-15:
  • the present invention does not specifically limit the method for preparing the pyrazolopyrimidine compound having the structure represented by formula (I). For example, it can be obtained by the following preparation method:
  • the above preparation method involves the Suzuki coupling reaction.
  • the reaction conditions of the coupling reaction are not particularly limited. Those skilled in the art can use the common knowledge in the field of organic synthesis and the specific examples provided in the Examples section of the present invention. Obtain suitable reaction conditions.
  • the second aspect of the present invention provides the pyrazolo pyrimidine compound having the structure represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as described in the first aspect of the present invention
  • pyrazolo pyrimidine compound having the structure represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof as described in the first aspect of the present invention
  • Forms, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, or prodrugs thereof are used in the preparation of drugs for the prevention and/or treatment of TRK kinase receptor-mediated diseases Applications.
  • the third aspect of the present invention provides a pharmaceutical composition
  • a pharmaceutical composition comprising a pharmaceutically acceptable carrier, excipient or diluent, and as an active ingredient the formula ( I)
  • the fourth aspect of the present invention provides the use of the pharmaceutical composition described in the third aspect of the present invention in the preparation of drugs for preventing and/or treating TRK kinase receptor-mediated diseases.
  • the fifth aspect of the present invention provides the pyrazolo pyrimidine compound having the structure represented by formula (I), or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof described in the first aspect of the present invention Forms, geometric isomers, tautomers, nitrogen oxides, hydrates, solvates, metabolites, or prodrugs thereof, or the pharmaceutical composition according to the third aspect of the present invention is prepared for prevention and/ Or the application of drugs to treat tumors.
  • the tumor is breast cancer, colorectal cancer, lung cancer, thyroid cancer, skin cancer, bone cancer, melanoma, leukemia, salivary gland tumor, neuroendocrine tumor, lymphoma, brain tumor, neuroblastoma, ovarian cancer , Pancreatic cancer, mesothelioma, esophageal cancer, lung sarcoma, medulloblastoma, glioblastoma, colon cancer, liver cancer, retinoblastoma, kidney cancer, bladder cancer, osteosarcoma, stomach cancer, uterine cancer, At least one of vulvar cancer, small bowel cancer, prostate cancer, cholangiocarcinoma, ureter cancer, adrenocortical cancer, or head and neck cancer.
  • Step 1) Add (R)4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-ylamino)ethyl)phenol (11.0mmol), 1,1-difluoro -2-iodoethane (16.5 mmol) and cesium carbonate (22 mmol) were added to a 200 mL pear-shaped flask, and DMF (50 mL) was added to it. Heat overnight in an oil bath (100°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 90%.
  • Step 2) Add (R)-N-(1-(2-(2,2-difluoroethoxy)-5-fluorophenyl)ethyl)pyrazolo[1,5-a]pyrimidine- 5-amine (9.8 mmol) was added to a 200 mL pear-shaped flask, and acetonitrile (50 mL) was added to it. At room temperature under magnetic stirring, N-iodosuccinimide (NIS, 14.85 mmol) was added. React at room temperature for 1 hour, and TLC monitors the completion of the reaction. Remove acetonitrile as much as possible under reduced pressure, dilute with 250 mL ethyl acetate, and transfer to a separatory funnel.
  • NMS N-iodosuccinimide
  • Step 3) Add (R)-N-(1-(2-(2,2-difluoroethoxy)-5-fluorophenyl)ethyl)-3-iodopyrazolo[1,5- a] Pyrimidine-5-amine (0.50mmol), 1-Boc-pyrazole-4-boronic acid pinacol ester (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis(triphenylphosphine)palladium (0.05 mmol) was added to a 100mL reaction tube, replaced with argon 3 times, 10mL anhydrous DMF and 2mL water were added.
  • the reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC. Cool to 50°C, filter with Celite, add water to the filtrate and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a black oily crude product. The crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 50%.
  • the preparation method is the same as in Example 1, except that 1-Boc-pyrazole-4-boronic acid pinacol ester is changed to 1-methylpyrazole-4-boronic acid pinacol ester.
  • Step 1) adopt the method of step 1 in embodiment 1.
  • Step 2) N-(1-(5-fluoro-2-(2-fluoroethoxy)phenyl)ethyl)-3-iodopyrazolo[1,5-a]pyrimidin-5-amine
  • the method of step 2 in Example 1 was used.
  • Step 3) The synthesis method is the same as step 3 in Example 1. The difference is that 1-Boc-pyrazole-4-boronic acid pinacol ester is replaced with 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2 -Dioxaborin-2-yl)-1H-pyrazole.
  • the synthesis method is the same as in Example 3, the difference is that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl ) -1H-pyrazole was replaced with 1-cyclopropylpyrazole-4-pinacol borate.
  • the preparation method is the same as in Example 1, except that 1-Boc-pyrazole-4-boronic acid pinacol ester is replaced with 4-(4,4,5,5-tetramethyl-1,3,2-di Oxborin-2-yl)-1H-pyrazole-1-ethanol.
  • the preparation method is the same as that in Example 3, except that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl )-1H-pyrazole is replaced with 1-(2,2-difluoroethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-pyrazole.
  • the preparation method is the same as in Example 1, except that 1-Boc-pyrazole-4-boronic acid pinacol ester is replaced with 2-[4-(4,4,5,5-tetramethyl-1,3, 2-Dioxaborolan-2-yl)pyrazol-1-yl]acetonitrile.
  • Example 8 2-(4-(5-((1-(2-(2,2-Difluoroethoxy)-5-fluorophenyl)ethyl)amino)pyrazolo[1,5- a) Preparation of pyrimidin-3-yl)-1H-pyrazopiperazin-1-yl)cyclopentane-1-ol (compound 8)
  • the preparation method is the same as in Example 8, except that 1,2-epoxycyclopentane is replaced with 1,2-epoxycyclohexane.
  • the preparation method is the same as in Example 1, the difference is that 1-Boc-pyrazole-4-boronic acid pinacol ester is replaced with 1-(tetrahydropyran-2-yl)-1H-pyrazole-4-boronic acid pin Which alcohol ester.
  • the preparation method is the same as that in Example 3, except that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl )-1H-pyrazole is replaced with 1-(tetrahydropyran-4-yl)-1H-pyrazole-4-boronic acid pinacol ester.
  • Step 1) Combine 1-(3,5-difluoro-2-hydroxyphenyl)ethan-1-one (29.0mmol), 1,1-difluoro-2-iodoethane (43.5mmol), cesium carbonate (58 mmol) was added to a 200 mL pear-shaped flask, and DMF (50 mL) was added to it. Heat overnight in an oil bath (80°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 78%.
  • Step 2) Combine 1-(2-(2,2-difluoroethoxy)-3,5-difluorophenyl)ethanone (22.3mmol), hydroxylamine hydrochloride (33.9mmol), cesium carbonate (44.6mmol) ) was added to a 200 mL pear-shaped flask, and methanol (50 mL) was added to it. The reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. The reaction solution was poured into water, filtered with suction, and dried to obtain a white solid with a yield of 98%.
  • Step 3) Combine 1-(2-(2,2-difluoroethoxy)-3,5-difluorophenyl)ethan-1-one oxime (21.4mmol), zinc powder (321mmol), chlorinated Ammonium (321 mmol) and acetic acid (321 mmol) were added to a 200 mL pear-shaped flask, and methanol (50 mL) was added to it. Heat overnight in an oil bath (80°C). The reaction was cooled to ambient temperature, neutralized, filtered through Celite, and the filtrate was added with water and extracted with ethyl acetate. The organic phase was washed twice with saturated brine, dried with anhydrous sodium sulfate, and concentrated to obtain a yellow liquid with a yield of 77%.
  • Step 4) Add 5-chloropyrazolo[1,5-a]pyrimidine (16.5mmol), 1-(2-(2,2-difluoroethoxy)-3,5-difluorophenyl) Ethyl-1-amine (16.5 mmol), anhydrous n-butanol (50 mL) and N,N-diisopropylethylamine (DIPEA, 49.5 mmol) were added to a 200 mL pear-shaped flask. Heat overnight in an oil bath (140°C).
  • the reaction was cooled to ambient temperature, and concentrated under reduced pressure to remove n-butanol and N,N-diisopropylethylamine (DIPEA) as much as possible to obtain a crude yellow oil.
  • DIPEA N,N-diisopropylethylamine
  • the crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 68%.
  • Step 5) N-(1-(2-(2,2-difluoroethoxy)-3,5-difluorophenyl)ethyl)pyrazolo[1,5-a]pyrimidine-5-
  • the amine (11.2 mmol) was added to a 200 mL pear-shaped flask, and acetonitrile (50 mL) was added to it.
  • acetonitrile 50 mL
  • N-iodosuccinimide N-iodosuccinimide (NIS, 13.4 mmol) was added. React at room temperature for 1 hour, and TLC monitors the completion of the reaction.
  • Step 6) Add N-(1-(2-(2,2-difluoroethoxy)-3,5-difluorophenyl)ethyl)-3-iodopyrazolo[1,5-a ]Pyrimidine-5-amine (0.50mmol), 1-Boc-pyrazole-4-boronic acid pinacol ester (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis(triphenylphosphine)palladium (0.05mmol) ) was added to a 100 mL reaction tube, replaced with argon 3 times, and 10 mL of anhydrous DMF and 2 mL of water were added.
  • the reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC. Cool to 50°C, filter with Celite, add water to the filtrate and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a black oily crude product. The crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 50%.
  • the preparation method is the same as in Example 14, except that 1-Boc-pyrazole-4-boronic acid pinacol ester is replaced with 1-methylpyrazole-4-boronic acid pinacol ester.
  • Step 1) Using the method of step 1 in Example 14, the difference is that 1-(3,5-difluoro-2-hydroxyphenyl)ethan-1-one is replaced with 1-(2-hydroxybenzene) ⁇ )ethane-1-one.
  • Step 2) adopt the method of step 2 in embodiment 14.
  • Step 3) adopt the method of step 3 in embodiment 14.
  • Step 4) adopt the method of step 4 in embodiment 14.
  • Step 5) adopt the method of step 5 in embodiment 14.
  • Step 6) The synthesis method is the same as step 6 in Example 14.
  • the preparation method is the same as in Example 16, except that 1-Boc-pyrazole-4-boronic acid pinacol ester is replaced with 1-methylpyrazole-4-boronic acid pinacol ester.
  • Step 1) Add 1-(5-fluoro-2-hydroxyphenyl)ethanone (32.4mmol), 4-methoxybenzyl bromide (38.9mmol), and cesium carbonate (48.6mmol) into a 200mL pear-shaped flask Acetonitrile (50 mL) was added to it. Heat overnight in an oil bath (80°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 78%.
  • Step 2) Combine 1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethanone (25.3mmol), hydroxylamine hydrochloride (30.4mmol), cesium carbonate (38.0mmol) Add to a 200 mL pear-shaped bottle, and add methanol (50 mL) to it. The reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. The reaction solution was poured into water, filtered with suction, and dried to obtain a white solid with a yield of 98%.
  • Step 3) Combine 1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethanone oxime (24.8mmol), zinc powder (372mmol), ammonium chloride (372mmol) , Acetic acid (372 mmol) was added to a 200 mL pear-shaped flask, and methanol (50 mL) was added to it. Heat overnight in an oil bath (80°C). The reaction was cooled to ambient temperature, neutralized, filtered through Celite, and the filtrate was added with water and extracted with ethyl acetate. The organic phase was washed twice with saturated brine, dried with anhydrous sodium sulfate, and concentrated to obtain a yellow liquid with a yield of 77%.
  • Step 4) Add 5-chloropyrazolo[1,5-a]pyrimidine (19.1mmol), 1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl Amine (19.1 mmol), anhydrous n-butanol (50 mL) and N,N-diisopropylethylamine (DIPEA, 38.2 mmol) were added to a 200 mL pear-shaped flask. Heat overnight in an oil bath (140°C).
  • DIPEA N,N-diisopropylethylamine
  • the reaction was cooled to ambient temperature, and concentrated under reduced pressure to remove n-butanol and N,N-diisopropylethylamine (DIPEA) as much as possible to obtain a crude yellow oil.
  • DIPEA N,N-diisopropylethylamine
  • the crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 68%.
  • Step 5) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)pyrazolo[1,5-a]pyrimidine-5- Amine (13.0 mmol) was added to a 200 mL pear-shaped flask, and acetonitrile (50 mL) was added to it. At room temperature under magnetic stirring, N-iodosuccinimide (NIS, 14.3 mmol) was added. React at room temperature for 1 hour, and TLC monitors the completion of the reaction. Remove the acetonitrile as much as possible under reduced pressure, dilute with 250mL ethyl acetate, and transfer to a separatory funnel.
  • NMS N-iodosuccinimide
  • Step 6) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)-3-iodopyrazolo[1,5-a] Pyrimidine-5-amine (0.77mmol), 1-methylpyrazole-4-boronic acid pinacol ester (1.16mmol), anhydrous potassium carbonate (2.00mmol), tetrakis(triphenylphosphine)palladium (0.08mmol) Add to a 100mL reaction tube, replace with argon 3 times, add 10mL anhydrous DMF and 2mL water. The reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC.
  • Step 7) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)-3-(1-methyl-1H-pyrazole- 3-yl)pyrazolo[1,5-a]pyrimidin-5-amine (0.22mmol) was added to a 200mL pear-shaped flask, dichloromethane (10mL) was added, and trifluoroacetic acid (3.3 mmol). The reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. Neutralize and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a white solid with a yield of 86%.
  • Example 19 4-Fluoro-2-(1-((3-(1-(2-hydroxycyclopentyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine- Preparation of 5-yl)amino)ethyl)phenol (compound a2-3)
  • Step 1) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)-3-(1H-pyrazole- 4-yl)pyrazolo[1,5-a]pyrimidin-5-amine (0.22mmol), 1,2-epoxycyclopentane (0.22mmol), anhydrous DMF (10mL) and cesium carbonate (0.6mmol) ). Heat to reflux overnight in an oil bath (100°C). The reaction was cooled to ambient temperature, and concentrated under reduced pressure to remove DMF as much as possible to obtain a yellow oily crude product. The crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 60%.
  • Step 2) adopt the method of step 7 in embodiment 18.
  • Example 20 4-Fluoro-2-(1-((3-(1-(2-hydroxycyclohexyl)-1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidine-5 -(Yl)amino)ethyl)phenol (compound a3-4) preparation
  • the preparation method is the same as in Example 19, except that 1,2-epoxycyclopentane is replaced with 1,2-epoxycyclohexane.
  • Step 1) Add 5-chloropyrazolo[1,5-a]pyrimidine (19.1mmol), (R)-1-(5-fluoro-2-((4-methoxybenzyl)oxy) Phenyl) ethylamine (19.1 mmol), anhydrous n-butanol (50 mL) and N,N-diisopropylethylamine (DIPEA, 38.2 mmol) were added to a 200 mL pear-shaped flask. Heat overnight in an oil bath (140°C).
  • DIPEA N,N-diisopropylethylamine
  • the reaction was cooled to ambient temperature, and concentrated under reduced pressure to remove n-butanol and N,N-diisopropylethylamine (DIPEA) as much as possible to obtain a crude yellow oil.
  • DIPEA N,N-diisopropylethylamine
  • the crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 68%.
  • Step 2) Add (R)-N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)pyrazolo[1,5-a] Pyrimidine-5-amine (13.0 mmol) was added to a 200 mL pear-shaped flask, dichloromethane (50 mL) was added, and trifluoroacetic acid (195 mmol) was added dropwise. The reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. Neutralize and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a white solid with a yield of 85%.
  • Step 3) Add (R)-4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-ylamino)ethyl)phenol (11.0mmol), difluoromonoiodomethane (16.5 mmol), cesium carbonate (22 mmol) was added to a 200 mL pear-shaped flask, and DMF (50 mL) was added to it. Heat overnight in an oil bath (100°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 91%.
  • Step 4) Add (R)-N-(1-(5-fluoro-2-difluoromethoxy-phenyl)ethyl)pyrazolo[1,5-a]pyrimidin-5-amine (9.8 mmol) was added to a 200 mL pear-shaped flask, and acetonitrile (50 mL) was added to it. At room temperature under magnetic stirring, N-iodosuccinimide (NIS, 14.85 mmol) was added. React at room temperature for 1 hour, and TLC monitors the completion of the reaction. Remove the acetonitrile as much as possible under reduced pressure, dilute with 250mL ethyl acetate, and transfer to a separatory funnel.
  • NMS N-iodosuccinimide
  • Step 5) Add (R)-N-(1-(5-fluoro-2-difluoromethoxy-phenyl)ethyl)-3-iodopyrazolo[1,5-a]pyrimidine-5 -Amine (0.50mmol), 1-methylpyrazole-4-boronic acid pinacol ester (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis (triphenylphosphine) palladium (0.05mmol) added to 100mL In the reaction tube, argon was replaced 3 times, and 10 mL of anhydrous DMF and 2 mL of water were added.
  • the reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC. Cool to 50°C, filter with Celite, add water to the filtrate and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a black oily crude product. The crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 52%.
  • Step 1) Combine 4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-ylamino)ethyl)phenol (11.0mmol), difluoromonoiodomethane (16.5mmol) , Cesium carbonate (22mmol) was added to a 200mL pear-shaped flask, and DMF (50mL) was added to it. Heat overnight in an oil bath (100°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 91%.
  • Step 2) Add N-(1-(-5-fluoro-2-difluoromethoxyphenyl)ethyl)pyrazolo[1,5-a]pyrimidin-5-amine (9.8mmol) to Into a 200 mL pear-shaped bottle, add acetonitrile (50 mL). At room temperature under magnetic stirring, N-iodosuccinimide (NIS, 14.85 mmol) was added. React at room temperature for 1 hour, and TLC monitors the completion of the reaction. Remove the acetonitrile as much as possible under reduced pressure, dilute with 250mL ethyl acetate, and transfer to a separatory funnel.
  • NMS N-iodosuccinimide
  • Step 3) Add N-(1-(5-fluoro-2-difluoromethoxyphenyl)ethyl)-3-iodopyrazolo[1,5-a]pyrimidin-5-amine (0.50mmol ), 4-(4,4,5,5-tetramethyl-1,3,2-dioxborin-2-yl)-1H-pyrazole-1-ethanol (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis(triphenylphosphine)palladium (0.05mmol) was added to a 100mL reaction tube, replaced with argon 3 times, 10mL anhydrous DMF and 2mL water were added.
  • the reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC. Cool to 50°C, filter with Celite, add water to the filtrate and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a black oily crude product. The crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 52%.
  • Step 1) Combine 4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-ylamino)ethyl)phenol (11.0mmol), iodoethane (16.5mmol), carbonic acid Cesium (22 mmol) was added to a 200 mL pear-shaped flask, and DMF (50 mL) was added to it. Heat overnight in an oil bath (100°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 88%.
  • Step 2) Add N-(1-(5-fluoro-2-ethoxyphenyl)ethyl)pyrazolo[1,5-a]pyrimidin-5-amine (9.8mmol) to 200mL
  • acetonitrile 50 mL
  • N-iodosuccinimide NMS, 14.85 mmol
  • Step 3) Add N-(1-(5-fluoro-2-ethoxyphenyl)ethyl)-3-iodopyrazolo[1,5-a]pyrimidin-5-amine (0.50mmol), 1-(Tetrahydropyran-4-yl)-1H-pyrazole-4-boronic acid pinacol ester (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis(triphenylphosphine)palladium (0.05mmol) ) was added to a 100 mL reaction tube, replaced with argon 3 times, and 10 mL of anhydrous DMF and 2 mL of water were added.
  • the reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC. Cool to 50°C, filter with Celite, add water to the filtrate and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and concentrated to obtain a black oily crude product. The crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 52%.
  • Example 24 Using a method similar to Example 24, except that iodoethane was replaced with iodoethanol, and 1-(tetrahydropyran-4-yl)-1H-pyrazole-4-boronic acid pinacol ester Replaced with 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborane-2-yl)-1H-pyrazole.
  • Example 24 A method similar to that of Example 24 was used, except that iodoethane was replaced with 2-iodoethyl methyl ether.
  • Example 21 A method similar to that in Example 21 was used, except that difluoromonoiodomethane was replaced with 1-fluoro-2-iodoethane.
  • Example 28 3-(1-(Difluoromethyl)-1H-pyrazol-4-yl)-N-(1-(5-fluoro-2-(2-fluoroethoxy)phenyl)ethyl Yl)pyrazolo[1,5-a]pyrimidin-5-amine (compound a11-18)
  • Example 24 A method similar to that in Example 24 was used, except that iodoethane was replaced with 1-fluoro-2-iodoethane and 1-(tetrahydropyran-4-yl)-1H-pyrazole- Replace 4-boronic acid pinacol ester with 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl )-1H-pyrazole.
  • the preparation method is the same as that in Example 28, except that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-pyrazole is replaced with 1-isopropyl-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)- 1H-pyrazole.
  • the preparation method is the same as that in Example 28, except that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-pyrazole was replaced with 1-cyclopropylpyrazole-4-pinacol boronic acid ester.
  • the preparation method is the same as that in Example 28, except that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-pyrazole is replaced with 4-(4,4,5,5-tetramethyl-1,3,2-dioxborin-2-yl)-1H-pyrazole-1-ethanol .
  • Example 33 3-(1-(2,2-Difluoroethyl)-1H-pyrazol-4-yl)-N-(1-(5-fluoro-2-(2-fluoroethoxy) (Phenyl) ethyl) pyrazolo[1,5-a]pyrimidin-5-amine (compound a16-23) preparation
  • the preparation method is the same as that in Example 28, except that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-pyrazole is replaced with 1-(2,2-difluoroethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborole Pentan-2-yl)-1H-pyrazole.
  • the preparation method is the same as that in Example 34, except that 1,2-epoxycyclopentane is replaced with 1,2-epoxycyclohexane.
  • Example 36 N-(1-(5-Fluoro-2-(2-fluoroethoxy)phenyl)ethyl)-3-(1-(tetrahydro-2H-pyran-4-yl)- Preparation of 1H-pyrazol-4-yl)pyrazolo[1,5-a]pyrimidin-5-amine (compound a19-26)
  • the preparation method is the same as that in Example 28, except that 1-(difluoromethyl)-4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolane- 2-yl)-1H-pyrazole was replaced with 1-(tetrahydropyran-4-yl)-1H-pyrazole-4-boronic acid pinacol ester.
  • the preparation method is the same as that of Example 27, except that 1-methylpyrazole-4-boronic acid pinacol ester is replaced with 1-(tetrahydropyran-2-yl)-4-(4,4,5, 5-Tetramethyl-1,3,2-dioxaborane-2-yl)-1H-pyrazole.
  • Example 38 1-(4-(5-((1-(5-Fluoro-2-(2-fluoroethoxy)phenyl)ethyl)amino)pyrazolo[1,5-a]pyrimidine -3-yl)-1H-pyrazol-1-yl)ethanone (compound a21-28) preparation
  • the preparation method is the same as in Example 38, except that acetyl chloride is replaced with methanesulfonyl chloride.
  • Example 21 A method similar to that in Example 21 was used, except that difluoromonoiodomethane was replaced with 2-iodo-1,1,1-trifluoroethane.
  • Example 21 A method similar to that in Example 21 was used, except that ethyl iodide was replaced with 2-iodo-1,1,1-trifluoroethane, and 1-methylpyrazole-4-boronic acid pinacol The ester was replaced by 4-(4,4,5,5-tetramethyl-1,3,2-dioxboran-2-yl)-1H-pyrazole-1-ethanol.
  • Example 24 A method similar to that of Example 24 was used, except that iodoethane was replaced with 3-fluoro-1-iodopropane.
  • Example 24 Using a method similar to Example 24, except that iodoethane was replaced with iodomethylcyclopropane, and 1-(tetrahydropyran-4-yl)-1H-pyrazole-4-boronic acid The ester is replaced with 1-methylpyrazole-4-boronic acid pinacol ester.
  • Example 24 Using a method similar to that of Example 24, except that iodoethane was replaced with 3-iodotetrahydrofuran, and 1-(tetrahydropyran-4-yl)-1H-pyrazole-4-boronic acid pinnacle The alcohol ester is replaced with 1-methylpyrazole-4-boronic acid pinacol ester.
  • Example 24 Using a method similar to Example 24, except that iodoethane was replaced with 4-iodomethyltetrahydropyran, and 1-(tetrahydropyran-4-yl)-1H-pyrazole- The 4-boronic acid pinacol ester was replaced with 1-methylpyrazole-4-boronic acid pinacol ester.
  • Step 1) Add 1-(5-fluoro-2-hydroxyphenyl)ethanone (32.4mmol), 4-methoxybenzyl bromide (38.9mmol), and cesium carbonate (48.6mmol) into a 200mL pear-shaped flask Acetonitrile (50 mL) was added to it. Heat overnight in an oil bath (80°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 78%.
  • Step 2) Combine 1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethanone (25.3mmol), hydroxylamine hydrochloride (30.4mmol), cesium carbonate (38.0mmol) Add to a 200 mL pear-shaped bottle, and add methanol (50 mL) to it. The reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. The reaction solution was poured into water, filtered with suction, and dried to obtain a white solid with a yield of 98%.
  • Step 3) Combine 1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethanone oxime (24.8mmol), zinc powder (372mmol), ammonium chloride (372mmol) , Acetic acid (372 mmol) was added to a 200 mL pear-shaped flask, and methanol (50 mL) was added to it. Heat overnight in an oil bath (80°C). The reaction was cooled to ambient temperature, neutralized, filtered through Celite, and the filtrate was added with water and extracted with ethyl acetate. The organic phase was washed twice with saturated brine, dried with anhydrous sodium sulfate, and concentrated to obtain a yellow liquid with a yield of 77%.
  • Step 4) Add 5-chloropyrazolo[1,5-a]pyrimidine (19.1mmol), 1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl Amine (19.1 mmol), anhydrous n-butanol (50 mL) and N,N-diisopropylethylamine (DIPEA, 38.2 mmol) were added to a 200 mL pear-shaped flask. Heat overnight in an oil bath (140°C).
  • DIPEA N,N-diisopropylethylamine
  • the reaction was cooled to ambient temperature, and concentrated under reduced pressure to remove n-butanol and N,N-diisopropylethylamine (DIPEA) as much as possible to obtain a crude yellow oil.
  • DIPEA N,N-diisopropylethylamine
  • the crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 68%.
  • Step 5) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)pyrazolo[1,5-a]pyrimidine-5- Amine (13.0 mmol) was added to a 200 mL pear-shaped flask, and acetonitrile (50 mL) was added to it. At room temperature, under magnetic stirring, add N-iodosuccinimide (NIS, 14.3 mmol). React at room temperature for 1 hour, and TLC monitors the completion of the reaction. Remove the acetonitrile as much as possible under reduced pressure, dilute with 250mL ethyl acetate, and transfer to a separatory funnel.
  • NMS N-iodosuccinimide
  • Step 6) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)-3-iodopyrazolo[1,5-a] Pyrimidine-5-amine (0.77mmol), 1-Boc-pyrazole-4-boronic acid pinacol ester (1.16mmol), anhydrous potassium carbonate (2.00mmol), tetrakis(triphenylphosphine)palladium (0.08mmol) Add to a 100mL reaction tube, replace with argon 3 times, add 10mL anhydrous DMF and 2mL water. The reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC.
  • Step 7) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)pyrazolo[1,5-a]pyrimidine-5- Amine (0.22 mmol) was added to a 200 mL pear-shaped flask, dichloromethane (10 mL) was added, and trifluoroacetic acid (3.3 mmol) was added dropwise. The reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. Neutralize and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a white solid with a yield of 85%.
  • Step 1) Add N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)pyrazolo[1,5-a]pyrimidine-5-
  • the amine 13.0 mmol
  • dichloromethane 50 mL
  • trifluoroacetic acid 195 mmol
  • the reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. Neutralize and extract with ethyl acetate.
  • the organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a white solid with a yield of 85%.
  • Step 2) Combine 4-fluoro-2-(1-(pyrazolo[1,5-a]pyrimidin-5-ylamino)ethyl)phenol (11.0mmol), methyl iodide (16.5mmol), cesium carbonate (22 mmol) was added to a 200 mL pear-shaped flask, and DMF (50 mL) was added to it. Heat overnight in an oil bath (100°C). The reaction was cooled to ambient temperature, and the crude product was purified by column chromatography to obtain a white solid with a yield of 90%.
  • Step 3) Add N-(1-(5-fluoro-2-methoxyphenyl)ethyl)pyrazolo[1,5-a]pyrimidin-5-amine (9.8mmol) to 200mL In a pear-shaped flask, acetonitrile (50 mL) was added. At room temperature under magnetic stirring, N-iodosuccinimide (NIS, 14.85 mmol) was added. React at room temperature for 1 hour, and TLC monitors the completion of the reaction. Remove the acetonitrile as much as possible under reduced pressure, dilute with 250mL ethyl acetate, and transfer to a separatory funnel.
  • NMS N-iodosuccinimide
  • Step 4) Add N-(1-(5-fluoro-2-methoxyphenyl)ethyl)-3-iodopyrazolo[1,5-a]pyrimidin-5-amine (0.50mmol), 1-Boc-pyrazole-4-boronic acid pinacol ester (0.75mmol), anhydrous potassium carbonate (2.00mmol), tetrakis (triphenylphosphine) palladium (0.05mmol) were added to the 100mL reaction tube, replaced with argon 3 times, add 10mL anhydrous DMF, 2ml water. The reaction was carried out at 100°C for 2 hours under argon atmosphere, and the completion of the reaction was monitored by TLC.
  • Step 1) Using the method of step 2 in Example 47, the difference is that methyl iodide is replaced with fluoromethyl iodide.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) Step 4 in Example 47 was used for the synthesis method.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with difluoromonoiodomethane.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) Step 4 in Example 47 was used for the synthesis method.
  • Step 1) The method of step 2 in Example 47 is adopted, except that methyl iodide is replaced with ethyl iodide.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) Step 4 in Example 47 was used for the synthesis method.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with 2-iodoethyl methyl ether.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) Step 4 in Example 47 was used for the synthesis method.
  • Step 1) Add 5-chloropyrazolo[1,5-a]pyrimidine (19.1mmol), (R)-1-(5-fluoro-2-((4-methoxybenzyl)oxy) Phenyl) ethylamine (19.1 mmol), anhydrous n-butanol (50 mL) and N,N-diisopropylethylamine (DIPEA, 38.2 mmol) were added to a 200 mL pear-shaped flask. Heat overnight in an oil bath (140°C).
  • DIPEA N,N-diisopropylethylamine
  • the reaction was cooled to ambient temperature, and concentrated under reduced pressure to remove n-butanol and N,N-diisopropylethylamine (DIPEA) as much as possible to obtain a crude yellow oil.
  • DIPEA N,N-diisopropylethylamine
  • the crude product was purified by column chromatography to obtain a pale yellow solid with a yield of 68%.
  • Step 2) Add (R)-N-(1-(5-fluoro-2-((4-methoxybenzyl)oxy)phenyl)ethyl)pyrazolo[1,5-a] Pyrimidine-5-amine (13.0 mmol) was added to a 200 mL pear-shaped flask, dichloromethane (50 mL) was added, and trifluoroacetic acid (195 mmol) was added dropwise. The reaction was carried out for 4 hours at room temperature under magnetic stirring conditions. Neutralize and extract with ethyl acetate. The organic phase was washed twice with saturated brine, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain a white solid with a yield of 85%.
  • Step 3) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with 1-fluoro-2-iodoethane.
  • Step 4) adopt the method of step 3 in embodiment 47.
  • Step 5 The synthesis method is the same as Step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with 2-iodo-1,1,1-trifluoroethane.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 is adopted, except that methyl iodide is replaced with propane iodide.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with 3-fluoro-1-iodopropane.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with ethyl iodide.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) Step 4 in Example 47 was used for the synthesis method.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with (iodomethyl)cyclopropane.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with cyclopentane iodide.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with 3-iodotetrahydrofuran.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 is used, except that methyl iodide is replaced with 4-iodomethyltetrahydropyran.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Step 1) The method of step 2 in Example 47 was used, except that methyl iodide was replaced with 2-(4-morpholine)ethyl bromide.
  • Step 2) adopt the method of step 3 in embodiment 47.
  • Step 3) The synthesis method is the same as step 4 in Example 47.
  • Test Example 1 In vitro biochemical level inhibition of protein kinase (PK) activity test
  • wild-type kinases such as TRKA, TRKB and TRKC
  • mutant kinases such as TRKA-G595R, TRKA-G667C, TRKA-F589L, TRKC-G623R, TRKC-G696A, etc., derived from Carna Biosciences 08-186, 08-187, 08-197
  • HTRF KinEASE TK kit (Cisbio 62TK0PEC); 384-well plate (Greiner); ATP (Life technologies PV3227), MgCl 2 (sigma); PHERAstar FS multifunctional microplate reader (BMG); low speed centrifugation Machine (StaiteXiangyi Company); Incubator (Binder Company).
  • the selected control compounds are the typical compound A disclosed in WO2007147647, the typical compound B disclosed in WO2007025540, and the comparative compound C and the comparative compound D.
  • Enzymatic buffer preparation Dilute 5 ⁇ Enzymatic buffer (from HTRF kit) with deionized water to 1.33 ⁇ , and add 1.33 ⁇ the final concentration of the corresponding components: 1.33mmol/L disulfide Thrreitol (DTT), 1.33mmol/L MnCl 2 , 6.65mmol/L MgCl 2 and 39.9nmol/L SEB;
  • the preparation of kinase working solution dilute TRKA, TRKB and TRKC to 2 ⁇ with 1.33 ⁇ enzyme buffer and the required concentrations are 0.404ng/ ⁇ L, 0.304ng/ ⁇ L and 0.236ng/ ⁇ L respectively;
  • TK Substrate Blr Dilute TK Substrate Blr with 1.33 ⁇ enzyme buffer to a mixture (from HTRF kit) and ATP (10mM) diluted to 4 ⁇ the required final concentration; TRKA, TRKB and TRKC
  • the final concentrations of ATP were 3.727 ⁇ mol/L, 2.56 ⁇ mol/L and 2.526 ⁇ mol/L.
  • the final concentration of TK Substrate-biotin is 0.2 ⁇ mol/L.
  • Enzyme reaction steps Add 4 ⁇ L of kinase working solution to each well of the low-volume 384 microwell plate, and at the same time add 4 ⁇ L of 1.33 ⁇ enzyme buffer as a negative control (Negative); add 2 ⁇ l of compound working solution to the well and add at the same time 2 ⁇ L of 8% DMSO aqueous solution was used as a zero compound concentration control (ie, positive control, Positive); incubated at 25°C for 5 min; added 2 ⁇ L of the substrate working solution to the well to start the enzyme reaction, and reacted at 37°C with shaking for 30 min.
  • HTRF reagent detection steps add 8 detection working solution to the well to stop the reaction; react at 25°C for 1 hour;
  • HTRF signal reading using PHERAstar FS reading detection signal, the corresponding settings of the instrument are as follows:
  • Number of flashes (Number of flashes) 200
  • IC 50 value take the logarithm of the compound concentration as the abscissa and the inhibition rate as the ordinate. In GraphPad Prism 5, fit a nonlinear curve: log(inhibitor) vs.response--Variable slope to find the enzyme activity test compound concentration at 50% inhibition i.e., IC 50.
  • the compounds provided by the present invention show excellent inhibitory activity against wild TRKA, TRKB and TRKC kinases, which is significantly better than typical compound A, typical compound B, comparative compound C and comparative compound D.
  • the compounds provided in some of the foregoing examples were administered to rats in the form of polyethylene glycol 400 aqueous solution (70%).
  • rats are given a dose of 5 mg/kg.
  • about 0.3 mL of blood samples were collected at 15, 30, 45 min, 1, 2, 4, 6, 8, 10, 24 h after administration, and placed in a heparinized Eppendorf tube, and stored temporarily on ice until centrifugation.
  • the whole blood was centrifuged at 8000 rpm for 5 minutes, and plasma was collected, and the plasma was transferred to a 96-well plate, and stored at -20°C for detection by LC-MS/MS.
  • the non-compartmental model of WinNonlin software was used to calculate the pharmacokinetic parameters of rats after administration.
  • Peak concentration C max adopt actual measured value
  • AUC0-t value calculated by the trapezoidal method
  • AUC 0- ⁇ AUC 0-t +Ct/ke
  • Ct is the plasma concentration at the last measurable time point
  • ke is the elimination rate constant
  • Absolute bioavailability F Dose iv *AUC 0-t, ig /Dose ig *AUC 0-t, iv ⁇ 100%.
  • Table 2 lists the pharmacokinetic parameters of the compound of the present invention in rats after intravenous administration. The results show that the compound of the present invention has good pharmacokinetic properties, including ideal clearance (CL), half-life (t 1/2 ), peak concentration (C max ) and exposure (AUC 0-t ).
  • Table 3 lists the pharmacokinetic parameters of the compound of the present invention in rats after oral administration. The results show that the compound of the present invention has good pharmacokinetic properties, including ideal clearance (CL), half-life (t 1/2 ), peak concentration (C max ), exposure (AUC 0-t ) and oral administration bioavailability.
  • Test Example 3 Inhibitory activity of the compound of the present invention on five TRK kinase mutants
  • Test Case 1 uses the same test method as Test Case 1.
  • Test Example 4 Anti-tumor activity of the compound of the present invention on a nude mouse xenograft tumor model
  • the pharmacodynamics of the compounds of the present invention were evaluated by a standard murine model of transplanted tumors.
  • Human non-small cell lung cancer H2228 was cultured and collected, and then subcutaneously inoculated into female nude mice aged 5-6 weeks (BALB/c, Shanghai Lingchang Biotechnology Co., Ltd.).
  • the tumor volume reached 100-150 mm 3
  • the animals were randomly divided into a solvent control group (70% PEG-400 in water) and a compound group (6 animals in each group).
  • Subsequent intragastric administration of the compounds in the examples (corresponding doses, dissolved in 70% PEG-400 in an aqueous solution) to animals starts anywhere from 0 to 7 days after tumor cell inoculation, and every day in the experiment Do it twice.
  • the experimental index is to investigate the effect of the compounds of the examples on tumor growth, and the specific index is T/C% or tumor inhibition rate TGI (%).
  • V 1/2 ⁇ a ⁇ b 2 where a and b represent length and width respectively.
  • T/C(%) (TT 0 )/(CC 0 ) ⁇ 100 where T and C are the tumor volume at the end of the experiment; T 0 and C 0 are the tumor volume at the beginning of the experiment.
  • TGI Tumor inhibition rate
  • TGI tumor inhibition rate
  • the tumor is smaller than the initial volume, that is, when T ⁇ T 0 or C ⁇ C 0 , it is defined as partial tumor regression (PR); if the tumor disappears completely, it is defined as complete tumor regression (CR).
  • PR partial tumor regression
  • CR complete tumor regression
  • the following BID means twice a day administration.
  • the compound of the present invention exhibits excellent anti-tumor activity on the human non-small cell lung cancer H2228 nude mouse xenograft tumor model.
  • compound 5 50mg/kg, BID ⁇ 8 significantly inhibited the growth of human non-small cell lung cancer H2228 subcutaneously transplanted tumors in nude mice.
  • the tumor inhibition rate was 182%, and 4/6 tumors completely resolved;
  • the drug was stopped from D8 to the end of the experiment (D14), the tumor inhibition rate was 155%, 3/6 animal tumors completely resolved, 2/6 animal tumors partially resolved.
  • Test Example 5 Anti-tumor activity of the compound of the present invention in nude mouse xenograft tumor model
  • the efficacy of the compound of the present invention was evaluated by a standard murine model of transplanted tumor.
  • Human non-small cell lung cancer H2228 was cultured and collected, and then subcutaneously inoculated into female nude mice aged 5-6 weeks (BALB/c, Shanghai Lingchang Biotechnology Co., Ltd.).
  • the tumor volume reached 100-150 mm 3
  • the animals were randomly divided into a solvent control group (70% PEG-400 in water) and a compound group (6 animals in each group).
  • Subsequent intragastric administration of the compounds of the examples (corresponding dose, dissolved in 70% PEG-400 in an aqueous solution) to animals starts from anywhere from 0 to 13 days after tumor cell inoculation, and every day in the test Do it once or twice.
  • the experimental index is to investigate the effect of the compounds of the examples on tumor growth, and the specific index is T/C% or tumor inhibition rate TGI (%).
  • V 1/2 ⁇ a ⁇ b 2 where a and b represent length and width respectively.
  • T/C(%) (TT 0 )/(CC 0 ) ⁇ 100 where T and C are the tumor volume at the end of the experiment; T 0 and C 0 are the tumor volume at the beginning of the experiment.
  • TGI Tumor inhibition rate
  • TGI tumor inhibition rate
  • the tumor is smaller than the initial volume, that is, when T ⁇ T 0 or C ⁇ C 0 , it is defined as partial tumor regression (PR); if the tumor disappears completely, it is defined as complete tumor regression (CR).
  • PR partial tumor regression
  • CR complete tumor regression
  • the compound of the present invention exhibits excellent anti-tumor activity in the human non-small cell lung cancer H2228 nude mouse xenograft tumor model.
  • compound b8-30 25mg/kg, BID ⁇ 14 significantly inhibited the growth of human non-small cell lung cancer H2228 subcutaneously transplanted tumors in nude mice.
  • the tumor inhibition rate was 95%, and the dose was increased from D8 To 50mg/kg, the tumor regressed, and to the end of the experiment (D14), the tumor inhibition rate was 127%, and 6/6 tumors regressed partially; compound b8-30 (50mg/kg, QD, QD ⁇ 11) had a negative effect on H2228.
  • the tumor inhibition rate of subcutaneously transplanted tumors in mice was 96% (D7). Starting from D8, the dose was increased to 100mg/kg, and the tumor subsided. By the end of the experiment (D14), the tumor inhibition rate was 110%, and 3/6 tumors partially resolved .
  • the pyrazolopyrimidine compound having the structure represented by formula (I) or its pharmaceutically acceptable salt, or its stereoisomer, geometric isomer, tautomer, nitrogen Oxides, hydrates, solvates, metabolites, or their prodrugs exhibit excellent inhibitory activity against TRK kinase, and at the same time, can exhibit good anti-tumor activity at the animal level.

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Abstract

本发明涉及生物医药领域，公开了吡唑并嘧啶化合物和药物组合物及其应用，该含氧取代吡唑并嘧啶化合物具有式(I)所示的结构。本发明提供的具有式(I)所示结构的含氧取代吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药对TRK激酶表现出优异的抑制活性，同时，在动物水平上表现出良好的抗肿瘤活性。

Description

吡唑并嘧啶化合物和药物组合物及其应用 技术领域
本发明涉及生物医药领域，具体涉及一种吡唑并嘧啶化合物、一种含有该吡唑并嘧啶化合物的药物组合物、前述吡唑并嘧啶化合物及药物组合物的应用。
背景技术
NTRK/TRK(Tropomyosin receptor kinase)为祌经营养因子酪氨酸受体，隶属于受体酪氨酸激酶家族。TRK家族主要包括3个成员，NTRK1/TRKA，NTRK2/TRKB和NTRK3/TRKC。完整的TRK激酶包括胞外区、跨膜区和胞内区三个部分。TRK激酶的胞外区与相应的配体结合之后，能够引起激酶构型变化，形成二聚体。TRK激酶的胞内区发生自体磷酸化从而激活自身的激酶活性，进而进一步激活下游的信号转导通路(如MAPK，AKT，PKC等)，产生相应的生物学功能；其中NGF(神经生长因子)结合TRKA，BDNF(衍生的神经营养因子)结合TRKB，以及NT3(神经营养因子3)结合TRKC。
TRK激酶在神经的发育过程中发挥重要的生理功能，包括神经元轴突的生长与功能维持、记忆的发生发展以及保护神经元免受伤害等等。同时，大量的研究表明TRK信号转导通路的活化与肿瘤的发生发展密切相关，在神经细胞瘤、前列腺癌、乳腺癌等中都发现了活化的TRK信号蛋白。
近几年来多种TRK融合蛋白的发现，更显示了其促进肿瘤发生的生物学功能。最早的TPM3-TRKA融合蛋白是在结肠癌细胞中发现的，在检测的临床病人中约有1.5％的发生率。后来在不同类型的临床肿瘤病人样本如肺癌，头颈癌、乳腺癌、甲状腺癌、神经胶质瘤等中发现了不同类型的TRK融合蛋白，如CD74-NTRK1，MPRIP-NTRK1，QKI-NTRK2，ETV6-NTRK3，BTB1-NTRK3等。这些不同的NTRK融合蛋白在不需要配体结合的情况下，自身处于高度活化的激酶活性状态，因而能够持续性的磷酸化下游的信号途径，诱导细胞增殖，促进肿瘤的发生、发展。
因此，近几年来，TRK融合蛋白己经成为一个有效的抗癌靶点和研究热点，例如WO2010048314、WO2012116217、WO2011146336、W02010033941、WO2018077246等均公开了具有不同结构类型的TRK激酶抑制剂。
此外，持续给药后出现的靶标突变是肿瘤产生耐药性的重要原因，近期临床上已经出现了TRK突变的病例，如TRKA G595R、G667C、G667S和F589L的突变(Russo M等.Cancer Discovery,2016,6(1)，36-44)、TRKC G623R和G696A的突变(Drilon A.等Annals of Oncology 2016,27(5),920-926)，而寻找新的TRK激酶抑制剂有望解决TRK突变引起的肿瘤耐药性问题。
另外，含氮芳杂环通常具有较好的成药性，典型的例子是ALK激酶抑制剂克唑替尼(Cui J.等.J.Med.Chem.2011,54,6342–6363)。WO2007147647和WO2007025540也分别公开了吡唑取代的吡唑并吡啶化合物和吡唑取代的咪唑并哒嗪化合物作为ALK激酶抑制剂及其在疾病治疗中的应用。
Figure PCTCN2020080198-appb-000001
发明内容
本发明的目的之一是为了提供一种新的具有优异的抗肿瘤活性的吡唑并嘧啶化合物。
现有技术中提供的典型化合物A和典型化合物B虽然对ALK激酶具有良好的抑制活性，但是，以典型化合物A和典型化合物B为代表的结构类似物对TRK激酶的抑制效果却并不佳。本发明的发明人通过大量的科学研究发现，具有本发明式(I)所示结构的吡唑并嘧啶化合物对TRK激酶表现出优异的抑制活性，且其抑制活性明显优于现有技术的典型化合物A和典型化合物B。更为重要的是，本发明的该类吡唑并嘧啶化合物在动物水平上的抗肿瘤活性同样明显优于典型化合物A和典型化合物B，从而表现出较现有技术更为优异的肿瘤治疗效果。
为了实现上述目的，本发明的第一方面提供一种具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药，
Figure PCTCN2020080198-appb-000002
其中，在式(I)中，
R 1、R 2、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
R 5选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、烷氧基取代的C 2-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基；
R 6选自H、C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；
R 7选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基。
本发明的第二方面提供前述第一方面中所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药在制备用于预防和/或治疗TRK激酶介导的疾病的药物中的应用。
本发明的第三方面提供一种药物组合物，其包含药学上可接受的载体、赋形剂或稀释剂，以及作为活性成分的本发明第一方面所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药。
本发明的第四方面提供本发明第三方面所述的药物组合物在制备用于预防和/或治疗TRK激酶受体介导的疾病的药物中的应用。
本发明的第五方面提供本发明第一方面中所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药，或者本发明第三方面中所述的物组合物在制备用于预防和/或治疗肿瘤的药物中的应用。
本发明提供的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药对TRK激酶表现出优异的抑制活性，同时，在动物水平上表现出良好的抗肿瘤活性。
具体实施方式
在本文中所披露的范围的端点和任何值都不限于该精确的范围或值，这些范围或值应当理解为包含接近这些范围或值的值。对于数值范围来说，各个范围的端点值之间、各个范围的端点值和单独的点值之间，以及单独的点值之间可以彼此组合而得到一个或多个新的数值范围，这些数值范围应被视为在本文中具体公开。
如前所述，本发明的第一方面提供一种吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药。
以下先对本发明中涉及的部分术语进行解释：
“C 1-12的烷基”表示碳原子总数为1-12的烷基，包括直链烷基、支链烷基或者环烷基，例如可以为碳原子总数为1、2、3、4、5、6、7、8、9、10、11或12的直链烷基、支链烷基或者环烷基，例如可以为甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、正戊基、异戊基、正己基、环丙基、甲基环丙基、乙基环丙基、环戊基、甲基环戊基、环己基等。针对“C 1-8的烷基”、“C 1-6的烷基”具有与此相似的解释，所不同的是，碳原子数不同。
“由1-6个卤素取代的C 1-12的烷基”表示碳原子总数为1-12的烷基，包括直链烷基、支链烷基和环烷基，并且该C 1-12的烷基中的1-6个H由选自卤素的卤原子取代，例如该C 1-12的烷基中的1、2、3、4、5或6个H由选自氟、氯、溴、碘中的任意一个或者多个卤原子取代，例如可以为三氟甲基、二氟甲基、一氟甲基、一氟乙基、二氟乙基、三氟乙基等。针对“由1-6个卤素取代的C 1-8的烷基”、“由1-6个卤素取代的C 1-6的烷基”具有与此相似的解释，所不同的是，碳原子数不同。
“羟基取代的C 1-12的烷基”表示碳原子总数为1-12的烷基，包括直链烷基、支链烷基或者环烷基，并且该C 1-12的烷基中的至少一个H由羟基取代。
“烷氧基取代的C 2-12的烷基”表示碳原子总数为2-12的基团，且该基团的结构式可以表示为-R 1OR 2，其中， R 1和R 2中的碳原子数之和为2-12，且R 1直接与本发明的式(I)所示结构的吡唑并嘧啶化合物中的苯氧基连接。
“氰基取代的C 2-12的烷基”表示碳原子总数为2-12的烷基，包括直链烷基、支链烷基或者环烷基，并且该C 2-12的烷基中的至少一个H由氰基取代，且该“氰基”中的碳原子计入该基团的碳原子总数中。
“含有1-3个选自N、O和S的杂原子的C 2-12的环烷基”表示碳原子总数为2-12的环烷基，且形成环的原子中的1-3个原子选自N、O和S的杂原子，并且形成环的原子上可以含有烷基取代基，该烷基取代基中含有的碳原子数包括在前述碳原子总数范围内。“含有1-3个选自N、O和S的杂原子的C 2-12的环烷基”例如可以为三元环、四元环、五元环、六元环、七元环、八元环、九元环、十元环、十一元环或十二元环，并且，环烷基中的H可以任意地被取代基取代或者未取代，若被取代，其中的取代基各自独立地选自卤素、羟基、硝基和巯基中的至少一种。
“C 2-12的酰基”表示碳原子总数为2-12的酰基，例如可以为乙酰基，丙酰基等。
“磺酰基”中可以含有C 1-6的烷基，所述磺酰基例如可以以-SO 2R 3，表示，其中的R 3可以为C 1-6的烷基。
在本发明所述式(I)所示结构的吡唑并嘧啶化合物中，优选R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；更优选地，R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基；进一步优选地，R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；且R 2为H或F。
根据另一种优选的实施方式，在本发明所述式(I)所示结构的吡唑并嘧啶化合物中，优选R 1、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；且R 2为卤素；更优选地，R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；且R 2为F；进一步优选地，R 1、R 3和R 4均为H；R 2为F。
在本发明所述式(I)所示结构的吡唑并嘧啶化合物中，优选R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；更优选地，R 5选自H、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基；进一步优选地，R 5选自由1-6个卤素取代的C 1-12的烷基；进一步优选地，R 5选自由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；进一步优选R 5为-CH 2CHF 2。
在本发明所述式(I)所示结构的吡唑并嘧啶化合物中，优选R 6选自H、C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；更优选地，R 6选自H、C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；进一步优选地，R 6选自H、C 1-6的烷基、羟基取代的C 1-6的烷基和卤素。
在本发明所述式(I)所示结构的吡唑并嘧啶化合物中，优选R 7选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基；更优选地，R 7选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基；进一步优选地，R 7选自H、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基、C 2-6的酰基、磺酰基；特别优选R 7为H。
特别地，本发明的发明人发现，当所述R 2为卤素时，本发明提供的式(I)所示结构的吡唑并嘧啶化合物对TRK激酶，特别是对突变的TRK激酶表现出更高的抑制活性；同时表现出更合理的药代动力学性质，更优异的体内抗肿瘤活性。
以下提供本发明的所述式(I)所示结构的吡唑并嘧啶化合物的几种优选的具体实施方式：
具体实施方式1：
在式(I)中，
R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
R 6选自H、C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
R 7选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基。
具体实施方式2：
在式(I)中，
R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基；
R 5选自H、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基；
R 6选自H、C 1-6的烷基、羟基取代的C 1-6的烷基和卤素；
R 7选自H、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基、C 2-6的酰基、磺酰基。
进一步优选情况下，在前述具体实施方式1和前述具体实施方式2中，更优选R 1、R 2、R 3、R 4和R 5中的至少一个基团中含有F原子；且R 5选自由1-6个卤素取代的C 1-12的烷基；进一步优选地，R 1、R 2、R 3、R 4和R 5中的至少一个基团中含有F原子；且R 5选自由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基。
具体实施方式3：
在式(I)中，
R 1、R 2、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
R 5为-CH 2CHF 2；
R 6选自C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；
R 7选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基。
具体实施方式4：
在式(I)中，
R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
R 5为-CH 2CHF 2；
R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
R 7选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 1-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基。
具体实施方式5：
R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
R 2为H或F；
R 5为-CH 2CHF 2；
R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
R 7选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基。
具体实施方式6：
式(I)所示结构的化合物选自以下化合物中的至少一种：
化合物1：
Figure PCTCN2020080198-appb-000003
化合物2：
Figure PCTCN2020080198-appb-000004
化合物3：
Figure PCTCN2020080198-appb-000005
化合物4：
Figure PCTCN2020080198-appb-000006
化合物5：
Figure PCTCN2020080198-appb-000007
化合物6：
Figure PCTCN2020080198-appb-000008
化合物7：
Figure PCTCN2020080198-appb-000009
化合物8：
Figure PCTCN2020080198-appb-000010
化合物9：
Figure PCTCN2020080198-appb-000011
化合物10：
Figure PCTCN2020080198-appb-000012
化合物11：
Figure PCTCN2020080198-appb-000013
化合物12：
Figure PCTCN2020080198-appb-000014
化合物13：
Figure PCTCN2020080198-appb-000015
化合物14：
Figure PCTCN2020080198-appb-000016
化合物15：
Figure PCTCN2020080198-appb-000017
化合物16：
Figure PCTCN2020080198-appb-000018
化合物17：
Figure PCTCN2020080198-appb-000019
进一步优选情况下，在前述具体实施方式1和前述具体实施方式2中，更优选R 7选自C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基；进一步优选地，R 7选自C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基、C 2-6的酰基、磺酰基。
具体实施方式7：
在式(I)中，
R 1、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
R 2为卤素；
R 5选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、烷氧基取代的C 2-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基；
R 6选自C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；
R 7选自C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基。
具体实施方式8：
在式(I)中，
R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
R 2为F；
R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
R 7选自C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 1-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基。
具体实施方式9：
在式(I)中，
R 1、R 3和R 4均为H；R 2为F；
R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
R 7选自C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基。
具体实施方式10：
式(I)所示结构的化合物选自以下化合物中的至少一种：
化合物a1-2：
Figure PCTCN2020080198-appb-000020
化合物a2-3：
Figure PCTCN2020080198-appb-000021
化合物a3-4：
Figure PCTCN2020080198-appb-000022
化合物a4-8：
Figure PCTCN2020080198-appb-000023
化合物a5-9：
Figure PCTCN2020080198-appb-000024
化合物a6-10：
Figure PCTCN2020080198-appb-000025
化合物a7-12：
Figure PCTCN2020080198-appb-000026
化合物a8-13：
Figure PCTCN2020080198-appb-000027
化合物a9-14：
Figure PCTCN2020080198-appb-000028
化合物a10-17：
Figure PCTCN2020080198-appb-000029
化合物a11-18：
Figure PCTCN2020080198-appb-000030
化合物a12-19：
Figure PCTCN2020080198-appb-000031
化合物a13-20：
Figure PCTCN2020080198-appb-000032
化合物a14-21：
Figure PCTCN2020080198-appb-000033
化合物a15-22：
Figure PCTCN2020080198-appb-000034
化合物a16-23：
Figure PCTCN2020080198-appb-000035
化合物a17-24：
Figure PCTCN2020080198-appb-000036
化合物a18-25：
Figure PCTCN2020080198-appb-000037
化合物a19-26：
Figure PCTCN2020080198-appb-000038
化合物a20-27：
Figure PCTCN2020080198-appb-000039
化合物a21-28：
Figure PCTCN2020080198-appb-000040
化合物a22-29：
Figure PCTCN2020080198-appb-000041
化合物a23-31：
Figure PCTCN2020080198-appb-000042
化合物a24-32：
Figure PCTCN2020080198-appb-000043
化合物a25-35：
Figure PCTCN2020080198-appb-000044
化合物a26-38：
Figure PCTCN2020080198-appb-000045
化合物a27-41：
Figure PCTCN2020080198-appb-000046
化合物a28-43：
Figure PCTCN2020080198-appb-000047
进一步优选情况下，在前述具体实施方式1和前述具体实施方式2中，更优选R 7为H。
具体实施方式11：
在式(I)中，
R 1、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
R 2为卤素；
R 5选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、烷氧基取代的C 2-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基；
R 6选自C 1-12的烷基、羟基取代的C 1-12的烷基和卤素。
具体实施方式12：
在式(I)中，
R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
R 2为F；
R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素。
具体实施方式13：
在式(I)中，
R 1、R 3和R 4均为H；R 2为F；
R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素。
具体实施方式14：
式(I)所示结构的化合物选自以下化合物中的至少一种：
化合物b1-1：
Figure PCTCN2020080198-appb-000048
化合物b2-5：
Figure PCTCN2020080198-appb-000049
化合物b3-6：
Figure PCTCN2020080198-appb-000050
化合物b4-7：
Figure PCTCN2020080198-appb-000051
化合物b5-11：
Figure PCTCN2020080198-appb-000052
化合物b6-15：
Figure PCTCN2020080198-appb-000053
化合物b7-16：
Figure PCTCN2020080198-appb-000054
化合物b8-30：
Figure PCTCN2020080198-appb-000055
化合物b9-33：
Figure PCTCN2020080198-appb-000056
化合物b10-34：
Figure PCTCN2020080198-appb-000057
化合物b11-36：
Figure PCTCN2020080198-appb-000058
化合物b12-37：
Figure PCTCN2020080198-appb-000059
化合物b13-39：
Figure PCTCN2020080198-appb-000060
化合物b14-40：
Figure PCTCN2020080198-appb-000061
化合物b15-42：
Figure PCTCN2020080198-appb-000062
化合物b16-44：
Figure PCTCN2020080198-appb-000063
需要说明的是，在前述各个具体化合物中，未标示手性中心具体构型的化合物，即表示为外消旋体。
本发明对制备具有式(I)所示结构的吡唑并嘧啶化合物的方法没有特别的限定，例如可以采用如下制备方法获得：
Figure PCTCN2020080198-appb-000064
上述制备方法中涉及的是Suzuki偶联反应，对所述偶联反应的反应条件没有特别的限定，本领域技术人员可以根据有机合成领域内的公知常识以及本发明实施例部分提供的具体实施例获得适宜的反应条件。
如前所述，本发明的第二方面提供本发明的第一方面中所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药在制备用于预防和/或治疗TRK激酶受体介导的疾病的药物中的应用。
如前所述，本发明的第三方面提供一种药物组合物，其包含药学上可接受的载体、赋形剂或稀释剂，以及作为活性成分的本发明第一方面所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药。
如前所述，本发明的第四方面提供本发明的第三方面中所述的药物组合物在制备用于预防和/或治疗TRK激酶受体介导的疾病的药物中的应用。
如前所述，本发明的第五方面提供本发明的第一方面中所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药，或者本发明第三方面所述的药物组合物在制备用于预防和/或治疗肿瘤的药物中的应用。
优选情况下，所述肿瘤为乳腺癌、大肠癌、肺癌、甲状腺癌、皮肤癌、骨癌、黑色素瘤、白血病、唾液腺肿瘤、神经内分泌肿瘤、淋巴瘤、脑肿瘤、成神经细胞瘤、卵巢癌、胰腺癌、间皮瘤、食管癌、肺肉瘤、成神经管细胞瘤、成胶质细胞瘤、结肠癌、肝癌、成视网膜细胞瘤、肾癌、膀胱癌、骨肉瘤、胃癌、子宫癌、外阴癌、小肠癌、前列腺癌、胆管癌、输尿管癌、肾上腺皮质癌或头颈部癌症中的至少一种。
以下将通过实施例对本发明进行详细描述。以下实施例中，在没有特别说明的情况下，使用的各种原料均来自商购，且为分析纯。
实施例1：(R)-N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物1)的制备
Figure PCTCN2020080198-appb-000065
步骤1)：将(R)4-氟-2-(1-(吡唑并[1,5-a]嘧啶-5-基氨基)乙基)苯酚(11.0mmol)，1,1-二氟-2-碘乙烷(16.5mmol)，碳酸铯(22mmol)加入到200mL的梨形瓶中，向其中加入DMF(50mL)。油浴(100℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率90％。
步骤2)：将(R)-N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(9.8mmol)，加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS，14.85mmol)。室温反应1h，TLC监测反应完毕。减压尽量除去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率67％。
步骤3)：将(R)-N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.50mmol)，1-Boc-吡唑-4-硼酸频哪醇酯(0.75mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.05mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2mL水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率50％。
1H NMR(400MHz,DMSO-d 6)δ12.71(s,1H),8.49(d,J＝7.6Hz,1H),8.08(s,1H),8.04(d,J＝6.8Hz,1H), 7.85(s,2H),7.16–6.99(m,3H),6.70–6.32(m,2H),5.63–5.44(m,1H),4.58–4.44(m,2H),1.47(d,J＝6.8Hz,3H).
实施例2：(R)-N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物2)的制备
制备方法同实施例1，不同之处在于将1-Boc-吡唑-4-硼酸频哪醇酯改为1-甲基吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝7.6Hz,1H),8.05(s,1H),8.02(d,J＝9.2Hz,1H),7.75(s,2H),7.23–6.99(m,3H),6.70–6.30(m,2H),5.57–5.46(m,1H),4.49(t,J＝14.4Hz,2H),3.85(s,3H),1.48(d,J＝6.8Hz,3H).
实施例3：N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1-(二氟甲基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物3)的制备
Figure PCTCN2020080198-appb-000066
步骤1)：采用实施例1中的步骤1的方法。
步骤2)：N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺的合成，采用实施例1中的步骤2的方法。
步骤3)：合成方法同实施例1中的步骤3。不同之处在于将1-Boc-吡唑-4-硼酸频哪醇酯替换为1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑。
1H NMR(400MHz,DMSO-d 6)δ8.53(d,J＝7.6Hz,1H),8.24(s,1H),8.19(s,1H),8.13(d,J＝6.0Hz,2H),7.99–7.53(m,1H),7.21–6.96(m,3H),6.64–6.30(m,2H),5.55–5.46(m,1H),4.45(ddq,J＝17.6,11.2,3.2Hz,2H),1.48(d,J＝6.8Hz,3H).
实施例4：3-(1-环丙基-1H-吡唑-4-基)-N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(化合物4)的制备
合成方法同实施例3,不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑替换为1-环丙基吡唑-4-硼酸片呐醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝7.6Hz,1H),8.04(s,1H),8.00(d,J＝6.8Hz,1H),7.81(s,1H),7.73(s,1H),7.25–6.96(m,3H),6.67–6.29(m,2H),5.55–5.44(m,1H),4.48(t,J＝14.4Hz,2H),3.77–3.54(m,1H),1.47(d,J＝6.8Hz,3H),1.08–0.96(d,J＝6.4Hz,4H).
实施例5：(R)-2-(4-(5-((1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)乙醇(化合物5)的制备
制备方法同实施例1，不同之处在于将1-Boc-吡唑-4-硼酸频哪醇酯替换为4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑-1-乙醇。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.04(s,1H),7.98(d,J＝6.8Hz,1H),7.77(d,J＝11.2Hz,2H),7.19–6.96(m,3H),6.67–6.30(m,2H),5.60–5.41(m,1H),4.94(t,J＝5.2Hz,1H),4.47(qd,J＝12.8,10.8,5.6Hz,2H),4.11(t,J＝5.6Hz,2H),3.73(q,J＝5.6Hz,2H),1.45(d,J＝6.8Hz,3H).
实施例6：N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1-(2,2-二氟乙基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物6)的制备
制备方法同实施例3，不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑替换为1-(2,2-二氟乙基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑。
1H NMR(400MHz,DMSO-d 6)δ8.50(d,J＝7.6Hz,1H),8.09(s,1H),8.02(d,J＝7.2Hz,1H),7.88(d,J＝7.6 Hz,2H),7.22–6.96(m,3H),6.70–6.14(m,3H),5.58–5.48(m,1H),4.59(dt,J＝15.2,9.2Hz,2H),4.46(dt,J＝14.4,3.6Hz,2H),1.48(d,J＝6.8Hz,3H).
实施例7：((R)-2-(4-(5-((1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)乙腈(化合物7)的制备
制备方法同实施例1，不同之处在于将1-Boc-吡唑-4-硼酸频哪醇酯替换为2-[4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吡唑-1-基]乙腈。
1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝7.6Hz,1H),8.09(s,1H),8.05(d,J＝7.2Hz,1H),7.91(d,J＝7.6Hz,2H),7.21–6.97(m,3H),6.67–6.30(m,2H),5.54–5.42(m,3H),4.61–4.35(m,2H),1.45(d,J＝7.2Hz,3H).
实施例8：2-(4-(5-((1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑哌嗪-1-基)环戊烷-1-醇(化合物8)的制备
Figure PCTCN2020080198-appb-000067
在茄形瓶中加入N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(0.22mmol)，1,2-环氧环戊烷(0.22mmol)，无水DMF(10mL)和碳酸铯(0.6mmol)。在油浴(100℃)中加热回流过夜。将反应冷却至环境温度，减压浓缩尽量除去DMF得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率60％。
1H NMR(400MHz,DMSO-d 6)δ8.46(d,J＝7.6Hz,1H),8.03(d,J＝1.2Hz,1H),7.95(t,J＝7.6Hz,1H),7.81(d,J＝10.0Hz,1H),7.77(s,1H),7.20–6.97(m,3H),6.67–6.31(m,2H),5.55–5.44(m,1H),5.07(dd,J＝6.4,4.8Hz,1H),4.60–4.36(m,2H),4.34–4.10(m,2H),2.22–2.06(m,1H),2.03–1.88(m,2H),1.78(p,J＝7.2Hz,2H),1.63–1.52(m,1H),1.45(d,J＝6.8Hz,3H).
实施例9：2-(4-(5-((1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑哌嗪-1-基)环己烷-1-醇(化合物9)的制备
制备方法同实施例8，不同之处在于将1,2-环氧环戊烷替换为1,2-环氧环己烷。
1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝7.6Hz,1H),8.05(s,1H),7.98(t,J＝7.6Hz,1H),7.84–7.74(m,2H),7.23–6.97(m,3H),6.72–6.29(m,2H),5.60–5.47(m,1H),4.79(dd,J＝11.6,5.2Hz,1H),4.59–4.40(m,2H),3.86–3.67(m,2H),2.07–1.64(m,5H),1.58–1.17(m,6H).
实施例10：N-((R)-1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1-(四氢-2H-吡喃-2-基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物10)的制备
制备方法同实施例1，不同之处在于将1-Boc-吡唑-4-硼酸频哪醇酯替换为1-(四氢吡喃-2-基)-1H-吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝7.6Hz,1H),8.12–8.06(m,1H),8.04–7.96(m,1H),7.93(s,1H),7.83(s,1H),7.20–6.94(m,3H),6.68–6.30(m,2H),5.54–5.41(m,1H),5.41–5.28(m,1H),4.45(td,J＝14.4,3.6Hz,2H),3.98–3.84(m,1H),3.70–3.58(m,1H),2.15–1.84(m,3H),1.74–1.63(m,1H),1.58–1.50(m,2H),1.45(d,J＝6.8Hz,3H).
实施例11：N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物11)的制备
制备方法同实施例3，不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑替换为1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.04(s,1H),7.96(d,J＝7.2Hz,1H),7.82(s,1H),7.76(s,1H),7.20–6.99(m,3H),6.66–6.32(m,2H),5.54–5.39(m,1H),4.57–4.26(m,3H),4.07–3.92(m,2H),3.57–3.44(m,2H),2.03–1.88(m,4H)，1.46(d,J＝6.8Hz,3H).
实施例12：(R)-1-(4-(5-((1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)乙酮(化合物12)的制备
在茄形瓶中加入N-(1-(5-氟-2-(2,2-二氟乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(0.26mmol)，乙酰氯(0.26mmol)，无水DCM(10mL)和三乙胺(0.52mmol)。在0℃下反应4h。减压浓缩得粗品，粗品经柱层析纯化得淡黄色固体，收率70％。
1H NMR(400MHz,DMSO-d 6)δ8.53(d,J＝7.6Hz,1H),8.41(s,1H),8.25(d,J＝8.0Hz,2H),8.19(d,J＝6.8Hz,1H),7.18–6.93(m,3H),6.68–6.32(m,2H),5.50(t,J＝7.2Hz,1H),4.65–4.36(m,2H),2.64(s,3H),1.45(d,J＝6.8Hz,3H).
实施例13：N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1-(甲基磺酰基)-1H-吡唑-4-基)吡唑并[1,5-A]嘧啶-5-胺(化合物13)的制备
Figure PCTCN2020080198-appb-000068
在茄形瓶中加入N-(1-(2-(2,2-二氟乙氧基)-5-氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(0.26mmol)，甲基磺酰氯(0.26mmol)，无水DCM(10mL)和三乙胺(0.52mmol)。在0℃下反应4h。减压浓缩得粗品，粗品经柱层析纯化得淡黄色固体，收率70％。
1H NMR(400MHz,DMSO-d 6)δ8.52(d,J＝7.6Hz,1H),8.31(d,J＝11.2Hz,2H),8.24(s,1H),8.13(d,J＝7.2Hz,1H),7.19–6.94(m,3H),6.63–6.27(m,2H),5.56–5.42(m,1H),4.58–4.32(m,2H),3.49(s,3H),1.46(d,J＝6.8Hz,3H).
实施例14：N-(1-(2-(2,2-二氟乙氧基)-3,5-二氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物14)的制备
Figure PCTCN2020080198-appb-000069
步骤1)：将1-(3,5-二氟-2-羟基苯基)乙-1-酮(29.0mmol)，1,1-二氟-2-碘乙烷(43.5mmol)，碳酸铯(58mmol)加入到200mL的梨形瓶中，向其中加入DMF(50mL)。油浴(80℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率78％。
步骤2)：将1-(2-(2,2-二氟乙氧基)-3,5-二氟苯基)乙酮(22.3mmol)，盐酸羟胺(33.9mmol)，碳酸铯(44.6mmol)加入到200mL的梨形瓶中，向其中加入甲醇(50mL)。在室温，磁力搅拌条件下，反应4h。将反应液倒入水中，抽滤，干燥，得到白色固体，收率98％。
步骤3)：将1-(2-(2,2-二氟乙氧基)-3,5-二氟苯基)乙-1-酮肟(21.4mmol)，锌粉(321mmol)，氯化铵(321mmol)，乙酸(321mmol)加入到200mL的梨形瓶中，向其中加入甲醇(50mL)。油浴(80℃)中加热过夜。将反应冷却至环境温度，中和，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黄色液体，收率77％。
步骤4)：将5-氯吡唑并[1,5-a]嘧啶(16.5mmol)，1-(2-(2,2-二氟乙氧基)-3,5-二氟苯基)乙-1-胺(16.5 mmol)，无水正丁醇(50mL)和N,N-二异丙基乙胺(DIPEA，49.5mmol)加入到200mL的梨形瓶中。在油浴(140℃)中加热过夜。将反应冷却至环境温度，减压浓缩尽量除去正丁醇和N，N-二异丙基乙胺(DIPEA)得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率68％。
步骤5)：N-(1-(2-(2,2-二氟乙氧基)-3,5-二氟苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(11.2mmol)，加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS，13.4mmol)。室温反应1h，TLC监测反应完毕。减压尽量出去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率67％。
步骤6)：将N-(1-(2-(2,2-二氟乙氧基)-3,5-二氟苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.50mmol)，1-Boc-吡唑-4-硼酸频哪醇酯(0.75mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.05mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2mL水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率50％。
1H NMR(400MHz,DMSO-d 6)δ12.71(s,1H),8.49(d,J＝7.6Hz,1H),8.07(s,1H),7.99(d,J＝6.8Hz,1H),7.87(s,2H),7.26–7.18(m,1H),7.07–7.01(m,1H),6.63–6.25(m,2H),5.56–5.50(m,1H),4.62–4.30(m,2H),1.49(d,J＝6.8Hz,3H).
实施例15：N-(1-(2-(2,2-二氟乙氧基)-3,5-二氟苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物15)的制备
制备方法同实施例14，不同之处在于将1-Boc-吡唑-4-硼酸频哪醇酯替换为1-甲基吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.02(d,J＝16.4Hz,2H),7.82(s,1H),7.68(s,1H),7.24(d,J＝9.6Hz,1H),7.02(d,J＝9.2Hz,1H),6.62–6.26(m,2H),5.59–5.39(m,1H),4.55–4.31(m,2H),3.84(s,3H),1.49(d,J＝6.8Hz,3H).
实施例16：N-(1-(2-(2,2-二氟乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物16)的制备
Figure PCTCN2020080198-appb-000070
步骤1)：采用实施例14中的步骤1的方法，不同之处在于将1-(3,5-二氟-2-羟基苯基)乙-1-酮替换为1-(2-羟基苯基)乙烷-1-酮。
步骤2)：采用实施例14中的步骤2的方法。
步骤3)：采用实施例14中的步骤3的方法。
步骤4)：采用实施例14中的步骤4的方法。
步骤5)：采用实施例14中的步骤5的方法。
步骤6)：合成方法同实施例14中步骤6。
1H NMR(400MHz,DMSO-d 6)δ12.68(s,1H),8.43(d,J＝7.6Hz,1H),8.02(s,1H),7.99(d,J＝7.2Hz,1H),7.83(s,2H),7.31(dd,J＝7.6,1.6Hz,1H),7.23–7.12(m,1H),7.06(d,J＝8.0Hz,1H),6.92(t,J＝7.6Hz,1H),6.67–6.28(m,2H),5.58–5.51(m,1H),4.50–4.40(m,2H),1.43(d,J＝6.4Hz,3H).
实施例17：N-(1-(2-(2,2-二氟乙氧基)苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物17)的制备
制备方法同实施例16，不同之处在于将1-Boc-吡唑-4-硼酸频哪醇酯替换为1-甲基吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.43(d,J＝7.2Hz,1H),7.98(m,2H),7.72(d,J＝7.2Hz,2H),7.31(m,1H),7.19(m,1H),7.11(m,1H),6.93(m,1H),6.71–6.23(m,2H),5.59–5.47(m,1H),4.55–4.38(m,2H),3.81(s,3H),1.44(d,J＝7.2Hz,3H).
实施例18：4-氟-2-(1-((3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-基)氨基)乙基)苯酚(化合物a1-2)的制备
Figure PCTCN2020080198-appb-000071
步骤1)：将1-(5-氟-2-羟基苯基)乙酮(32.4mmol)，4-甲氧基溴苄(38.9mmol)，碳酸铯(48.6mmol)加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。油浴(80℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率78％。
步骤2)：将1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙酮(25.3mmol)，盐酸羟胺(30.4mmol)，碳酸铯(38.0mmol)加入到200mL的梨形瓶中，向其中加入甲醇(50mL)。在室温，磁力搅拌条件下，反应4h。将反应液倒入水中，抽滤，干燥，得到白色固体，收率98％。
步骤3)：将1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙酮肟(24.8mmol)，锌粉(372mmol)，氯化铵(372mmol)，乙酸(372mmol)加入到200mL的梨形瓶中，向其中加入甲醇(50mL)。油浴(80℃)中加热过夜。将反应冷却至环境温度，中和，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黄色液体，收率77％。
步骤4)：将5-氯吡唑并[1,5-a]嘧啶(19.1mmol)，1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙胺(19.1mmol)，无水正丁醇(50mL)和N，N-二异丙基乙胺(DIPEA，38.2mmol)加入到200mL的梨形瓶中。在油浴(140℃)中加热过夜。将反应冷却至环境温度，减压浓缩尽量除去正丁醇和N，N-二异丙基乙胺(DIPEA)得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率68％。
步骤5)：将N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(13.0mmol)，加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS，14.3mmol)。室温反应1h，TLC监测反应完毕。减压尽量出去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率67％。
步骤6)：将N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.77mmol)，1-甲基吡唑-4-硼酸频哪醇酯(1.16mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.08mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2mL水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率52％。
步骤7)：将N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)-3-(1-甲基-1H-吡唑-3-基)吡唑并[1,5-a]嘧啶-5-胺(0.22mmol)，加入到200mL的梨形瓶中，向其中加入二氯甲烷(10mL)，滴加三氟乙酸(3.3mmol)。在室温，磁力搅拌条件下，反应4h。中和并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，柱层析纯化得白色固体，收率86％。
1H NMR(400MHz,DMSO-d 6)δ9.68–9.65(m,1H),8.44(d,J＝7.6Hz,1H),8.02(s,1H),7.96(d,J＝6.8Hz,1H),7.84(s,1H),7.75(s,1H),7.04–6.77(m,3H),6.33(d,J＝7.6Hz,1H),5.44(d,J＝7.2Hz,1H),3.84(s,3H),1.43(d,J＝6.8Hz,3H).
实施例19：4-氟-2-(1-((3-(1-(2-羟基环戊基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-基)氨基)乙基)苯酚(化合物a2-3)的制备
Figure PCTCN2020080198-appb-000072
步骤1)：在茄形瓶中加入N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(0.22mmol)，1,2-环氧环戊烷(0.22mmol)，无水DMF(10mL)和碳酸铯(0.6mmol)。在油浴(100℃)中加热回流过夜。将反应冷却至环境温度，减压浓缩尽量除去DMF得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率60％。
步骤2)：采用实施例18中的步骤7的方法。
1H NMR(400MHz,DMSO-d 6)δ9.60(d,J＝16.4Hz,1H),8.43(d,J＝7.6Hz,1H),8.03(d,J＝2.0Hz,1H),7.96–7.86(m,2H),7.82(s,1H),7.04–6.94(m,1H),6.88–6.77(m,2H),6.33(d,J＝7.6Hz,1H),5.54–5..44(m,1H),5.09–5..02(m,1H),4.42–4.14(m,2H),2.21–2.09(m,2H),2.02–1.87(m,2H),1.83–1.73(m,2H),1.63–1.54(m,2H),1.43(d,J＝6.8Hz,3H).
实施例20：4-氟-2-(1-((3-(1-(2-羟基环己基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-基)氨基)乙基)苯酚(化合物a3-4)的制备
制备方法同实施例19，不同之处在于将1,2-环氧环戊烷替换为1,2-环氧环己烷。
1H NMR(400MHz,DMSO-d 6)δ9.59(d,J＝18.0Hz,1H),8.43(d,J＝7.6Hz,1H),8.01(d,J＝2.8Hz,1H),7.96–7.85(m,2H),7.80(s,1H),7.06–6.95(m,1H),6.82(dt,J＝6.4,1.6Hz,2H),6.42–6.22(m,1H),5.54–5.41(m,1H),4.84–4.64(m,1H),3.92–3.64(m,2H),2.03–1.65(m,6H),1.52–1.27(m,7H).
实施例21：(R)-N-(1-(2-(二氟甲氧基)-5-氟苯基)乙基)-3-(1-甲基-1H-吡唑-3-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a4-8)的制备
Figure PCTCN2020080198-appb-000073
步骤1)：将5-氯吡唑并[1,5-a]嘧啶(19.1mmol)，(R)-1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙胺(19.1mmol)，无水正丁醇(50mL)和N，N-二异丙基乙胺(DIPEA，38.2mmol)加入到200mL的梨形瓶中。在油浴(140℃)中加热过夜。将反应冷却至环境温度，减压浓缩尽量除去正丁醇和N，N-二异丙基乙胺(DIPEA)得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率68％。
步骤2)：将(R)-N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(13.0mmol)，加入到200mL的梨形瓶中，向其中加入二氯甲烷(50mL)，滴加三氟乙酸(195mmol)。在室温，磁力搅拌条件下，反应4h。中和并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，柱层析纯化得白色固体，收率85％。
步骤3)：将(R)-4-氟-2-(1-(吡唑并[1,5-a]嘧啶-5-基氨基)乙基)苯酚(11.0mmol)，二氟一碘甲烷(16.5mmol)，碳酸铯(22mmol)加入到200mL的梨形瓶中，向其中加入DMF(50mL)。油浴(100℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率91％。
步骤4)：将(R)-N-(1-(5-氟-2-二氟甲氧基-苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(9.8mmol)，加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS，14.85mmol)。室温反应1h，TLC监测反应完毕。减压尽量出去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率67％。
步骤5)：将(R)-N-(1-(5-氟-2-二氟甲氧基-苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.50mmol)，1-甲基吡唑-4-硼酸频哪醇酯(0.75mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.05mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2mL水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率52％。
1H NMR(600MHz,DMSO-d 6)δ8.48(s,1H),8.07(d,J＝6.0Hz,2H),7.80(s,1H),7.71(s,2H),7.39–7.07(m,3H),6.34(s,1H),5.52–5.39(m,1H),3.83(s,3H),1.47d,J＝6.6Hz,3H).
实施例22：2-(4-(5-((1-(2-(二氟甲氧基)-5-氟苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑哌嗪-1-基)乙-1-醇(化合物a5-9)的制备
步骤1)：将4-氟-2-(1-(吡唑并[1,5-a]嘧啶-5-基氨基)乙基)苯酚(11.0mmol)，二氟一碘甲烷(16.5mmol)，碳酸铯(22mmol)加入到200mL的梨形瓶中，向其中加入DMF(50mL)。油浴(100℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率91％。
步骤2)：将N-(1-(-5-氟-2-二氟甲氧基苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(9.8mmol)，加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS，14.85mmol)。室温反应1h，TLC监测反应完毕。减压尽量出去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率65％。
步骤3)：将N-(1-(5-氟-2-二氟甲氧基苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.50mmol)，4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑-1-乙醇(0.75mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.05mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2mL水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率52％。
1H NMR(400MHz,DMSO-d6)δ8.50(d,J＝7.6Hz,1H),8.07(s,1H),8.06(s,1H),7.86(s,1H),7.76(s,1H),7.55–7.10(m,4H),6.37(d,J＝7.6Hz,1H),5.49(t,J＝7.2Hz,1H),4.95–4.88(m,1H),4.15(t,J＝6.0Hz,2H),3.77(q,J＝5.6Hz,2H),1.50(d,J＝7.2Hz,3H).
实施例23：N-(1-(2-(二氟甲氧基)-5-氟苯基)乙基)-3-(1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a6-10)的制备
采用与实施例22相似的方法，所不同的是，将4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑-1-乙醇替换为1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.50(d,J＝7.6Hz,1H),8.08(s,1H),8.06(s,1H),7.90(s,1H),7.78(s,1H),7.54–7.11(m,4H),6.38(d,J＝7.6Hz,1H),5.49(t,J＝7.0Hz,1H),4.36(dt,J＝10.0,4.4Hz,1H),4.03(dt,J＝11.2,3.2Hz,2H),3.50(dp,J＝11.2,4.4Hz,2H),1.98(td,J＝8.8,7.2,3.6Hz,4H),1.50(d,J＝6.8Hz,3H).
实施例24：N-(1-(2-乙氧基-5-氟苯基)乙基)-3-(1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a7-12)的制备
步骤1)：将4-氟-2-(1-(吡唑并[1,5-a]嘧啶-5-基氨基)乙基)苯酚(11.0mmol)，碘乙烷(16.5mmol)，碳酸铯(22mmol)加入到200mL的梨形瓶中，向其中加入DMF(50mL)。油浴(100℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率88％。
步骤2)：将N-(1-(5-氟-2-乙氧基苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(9.8mmol)，加入到200mL的 梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS，14.85mmol)。室温反应1h，TLC监测反应完毕。减压尽量出去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率68％。
步骤3)：将N-(1-(5-氟-2-乙氧基苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.50mmol)，1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯(0.75mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.05mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2mL水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率52％。
1H NMR(400MHz,DMSO-d6)δ8.48(d,J＝7.2Hz,1H),8.06(s,1H),7.98(d,J＝7.0Hz,1H),7.87(s,1H),7.82(s,1H),7.15–6.92(m,3H),6.38(d,J＝7.2Hz,1H),5.61–5.44(m,1H),4.44–3.92(m,5H),3.59–3.43(m,2H),1.96(s,4H),1.45(d,J＝7.2Hz,6H).
实施例25：2-(2-(1-((3-(1-(二氟甲基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-基)氨基)乙基)-4-氟苯氧基)乙醇(化合物a8-13)的制备
Figure PCTCN2020080198-appb-000074
采用与实施例24相似的方法，所不同的是，将碘乙烷替换为碘乙醇，以及将1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯替换为1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑。
1H NMR(400MHz,DMSO-d 6)δ8.52(d,J＝7.6Hz,1H),8.29(s,1H),8.19(s,1H),8.16(s,1H),8.09(d,J＝7.2Hz,1H),7.82(t,J＝59.6Hz,1H),7.12–6.91(m,3H),6.41(d,J＝7.6Hz,1H),5.59–5.46(m,1H),5.00(t,J＝5.6Hz,1H),4.23–4.03(m,2H),3.90–3.80(m,2H),1.48(d,J＝6.8Hz,3H).
实施例26：N-(1-(5-氟-2-(2-甲氧基乙氧基)苯基)乙基)-3-(1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-基)吡唑并[1，5-a]嘧啶-5-胺(化合物a9-14)的制备
Figure PCTCN2020080198-appb-000075
采用与实施例24相似的方法，所不同的是，将碘乙烷替换为2-碘乙基甲醚。
1H NMR(400MHz,DMSO-d 6)δ8.45(d,J＝7.6Hz,1H),8.03(s,1H),7.90(d,J＝7.2Hz,1H),7.82(s,1H),7.77(s,1H),7.08–6.94(m,3H),6.35(d,J＝7.6Hz,1H),5.54–5.43(m,1H),4.39–4.13(m,3H),4.03–3.94(m,2H),3.81–3.69(m,2H),3.53–3.44(m,2H),3.33(s,3H),1.99–1.83(m,4H),1.45(d,J＝6.8Hz,3H).
实施例27：(R)-N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a10-17)的制备
采用与实施例21相似的方法，所不同的是，将二氟一碘甲烷替换为1-氟-2-碘乙烷。
1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝7.2Hz,1H),8.05(s,1H),8.01(d,J＝7.2Hz,1H),7.76(s,2H),7.17–6.95(m,3H),6.38(d,J＝7.6Hz,1H),5.61–5.49(m,1H),4.99–4.90(m,1H),4.87–4.78(m,1H),4.51–4.32(m,2H),3.84(s,3H),1.47(d,J＝6.8Hz,3H).
实施例28：3-(1-(二氟甲基)-1H-吡唑-4-基)-N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(化合物a11-18)的制备
Figure PCTCN2020080198-appb-000076
采用与实施例24相似的方法，所不同的是，将碘乙烷替换为1-氟-2-碘乙烷，以及将1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯替换为1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑。
1H NMR(400MHz,DMSO-d 6)δ8.53(d,J＝7.2Hz,1H),8.26(s,1H),8.19(s,1H),8.12(d,J＝8.0Hz,2H),7.80(t,J＝59.2Hz,1H),7.05(m,3H),6.43(d,J＝7.6Hz,1H),5.58–5.49(m,1H),4.96–4.85(m,1H),4.83–4.74(m,1H),4.48–4.37(m,1H),4.38–4.30(m,1H),1.48(d,J＝6.8Hz,3H).
实施例29：(R)-2-(4-(5-((1-(5-氟-2-(2-氟乙氧基)苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)乙腈(化合物a12-19)的制备
采用与实施例21相似的方法，所不同的是，分别将二氟一碘甲烷替换为1-氟-2-碘乙烷，将1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯替换为2-[4-(4,4,5,5-四甲基-1,3,2-二氧硼戊环-2-基)吡唑-1-基]乙腈。
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝7.2Hz,1H),8.08(s,1H),8.04(s,1H),7.91(d,J＝8.0Hz,2H),7.20–6.93(m,3H),6.37(d,J＝7.2Hz,1H),5.59–5.50(m,1H),5.47–5.40(m,2H),4.96–4.87(m,1H),4.84–4.74(m,1H),4.48–4.30(m,2H),1.45(d,J＝6.8Hz,3H).
实施例30：N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1-异丙基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a13-20)的制备
制备方法同实施例28，不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑替换为1-异丙基-4-(4,4,5,5-四甲基-1,3,2-二氧杂戊硼烷-2-基)-1H-吡唑。
1H NMR(400MHz,DMSO-d6)δ8.45(d,J＝7.6Hz,1H),8.01(s,1H),7.91(d,J＝7.2Hz,1H),7.81(s,1H),7.73(s,1H),7.13–6.94(m,3H),6.35(d,J＝7.6Hz,1H),5.55–5.42(m,1H),4.89(t,J＝4.0Hz,1H),4.77(t,J＝4.0Hz,1H),4.49–4.28(m,3H),1.45(d,J＝6.8Hz,3H),1.40(d,J＝6.8Hz,6H).
实施例31：3-(1-环丙基-1H-吡唑-4-基)-N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(化合物a14-21)的制备
制备方法同实施例28，不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑替换为1-环丙基吡唑-4-硼酸片呐醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝7.6Hz,1H),8.04(s,1H),7.99(d,J＝6.8Hz,1H),7.81(s,1H),7.74(s,1H),7.17–6.97(m,3H),6.38(d,J＝7.6Hz,1H),5.57–5.46(m,1H),4.97–4.89(m,1H),4.84–4.78(m,1H),4.51–4.31(m,2H),3.71–3.60(m,1H),1.47(d,J＝6.8Hz,3H),1.06–0.95(m,4H).
实施例32：2-(4-(5-((1-(5-氟-2-(2-氟乙氧基)苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)乙醇(化合物a15-22)的制备
制备方法同实施例28，不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑替换为4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑-1-乙醇。
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝7.6Hz,1H),8.05(s,1H),7.98(d,J＝7.2Hz,1H),7.80(s,1H),7.77(s,1H),7.11–7.00(m,3H),6.37(d,J＝7.6Hz,1H),5.60–5.51(m,1H),4.96–4.91(m,1H),4.84–4.79(m,1H),4.54–4.31(m,2H),4.11(t,J＝5.6Hz,2H),3.73(t,J＝5.6Hz,2H),1.47(d,J＝6.8Hz,3H).
实施例33：3-(1-(2,2-二氟乙基)-1H-吡唑-4-基)-N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(化合物a16-23)的制备
制备方法同实施例28，不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑替换为1-(2,2-二氟乙基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑。
1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝7.6Hz,1H),8.08(s,1H),8.01(d,J＝7.2Hz,1H),7.89(s,1H),7.87(s,1H),7.19–6.96(m,3H),6.55–6.19(m,2H),5.60–5.50(m,1H),4.92(t,J＝4.0Hz,1H),4.80(t,J＝3.6Hz,1H),4.67–4.52(m,2H),4.42(d,J＝4.8Hz,1H),4.35(d,J＝4.8Hz,1H),1.47(d,J＝6.8Hz,3H).
实施例34：2-(4-(5-((1-(5-氟-2-(2-氟乙氧基)苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)环戊烷-1-醇(化合物a17-24)的制备
Figure PCTCN2020080198-appb-000077
在茄形瓶中加入N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(0.22mmol)，1,2-环氧环戊烷(0.22mmol)，无水DMF(10mL)和碳酸铯(0.6mmol)。在油浴(100℃)中加热回流过夜。将反应冷却至环境温度，减压浓缩尽量除去DMF得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率65％。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.04(s,1H),7.97(s,1H),7.83(d,J＝5.6Hz,1H),7.79(s,1H),7.15–6.97(m,3H),6.37(d,J＝7.6Hz,1H),5.58–5.49(m,1H),5.07(t,J＝5.6Hz,1H),4.97–4.89(m,1H),4.84–4.77(m,1H),4.51–4.15(m,4H),2.21–2.10(m,1H),2.02–1.90(m,2H),1.85–1.73(m,2H),1.64–1.54(m,1H),1.47(d,J＝6.8Hz,3H).
实施例35：2-(4-(5-((1-(5-氟-2-(2-氟乙氧基)苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)环己烷-1-醇(化合物a18-25)的制备
制备方法同实施例34，不同之处在于将1,2-环氧环戊烷替换为1,2-环氧环己烷。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.03(s,1H),7.95(d,J＝7.2Hz,1H),7.79(s,1H),7.75(d,J＝2.4Hz,1H),7.16–6.97(m,3H),6.37(d,J＝7.6Hz,1H),5.59–5.48(m,1H),4.98–4.69(m,3H),4.51–4.31(m,2H),3.86–3.66(m,2H),2.07–1.67(m,5H),1.47(d,J＝7.2Hz,3H),1.40–1.24(m,3H).
实施例36：N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-基)吡唑并[1，5-a]嘧啶-5-胺(化合物a19-26)的制备
制备方法同实施例28，不同之处在于将1-(二氟甲基)-4-(4,4,5,5-四甲基-1,3,2-二氧硼杂环戊烷-2-基)-1H-吡唑替换为1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.46(d,J＝7.8Hz,1H),8.04(s,1H),7.94(d,J＝7.2Hz,1H),7.83(s,1H),7.78(s,1H),7.16–6.94(m,3H),6.38(d,J＝7.6Hz,1H),5.59–5.41(m,1H),4.91(dt,J＝4.8,2.4Hz,1H),4.79(dt,J＝4.4,2.4Hz,1H),4.51–4.25(m,3H),4.00(dq,J＝10.8,3.2Hz,2H),3.49(dp,J＝10.8,3.6Hz,2H),1.99–1.81(m,4H),1.47(d,J＝6.8Hz,3H).
实施例37：N-((R)-1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1-(四氢-2H-吡喃-2-基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a20-27)的制备
制备方法同实施例27，不同之处在于将1-甲基吡唑-4-硼酸频哪醇酯替换为1-(四氢吡喃-2-基)-4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑。
1H NMR(400MHz,DMSO-d 6)δ8.49(d,J＝7.6Hz,1H),8.09(s,1H),8.00(d,J＝6.8Hz,1H),7.96(s,1H),7.85(s,1H),7.15–6.96(m,3H),6.38(d,J＝7.6Hz,1H),5.57–5.47(m,1H),5.39–5.32(m,1H),4.96–4.91(m,1H),4.84–4.78(m,1H),4.46–4.41(m,1H),4.39–4.34(m,1H),3.99–3.91(m,1H),3.71–3.62(m,1H),2.16–1.88(m,3H),1.78–1.68(m,1H),1.61–1.54(m,2H),1.47(d,J＝6.8Hz,3H).
实施例38：1-(4-(5-((1-(5-氟-2-(2-氟乙氧基)苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H- 吡唑-1-基)乙酮(化合物a21-28)的制备
Figure PCTCN2020080198-appb-000078
在茄形瓶中加入N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(0.26mmol)，乙酰氯(0.26mmol)，无水DCM(10mL)和三乙胺(0.52mmol)。在0℃下反应4h。减压浓缩得粗品，粗品经柱层析纯化得淡黄色固体，收率70％。
1H NMR(400MHz,DMSO-d 6)δ8.54(d,J＝7.6Hz,1H),8.47(s,1H),8.29(s,1H),8.27(s,1H),8.16(d,J＝7.2Hz,1H),7.15–6.96(m,3H),6.43(d,J＝7.6Hz,1H),5.62–5.53(m,1H),4.99–4.90(m,1H),4.87–4.78(m,1H),4.57–4.32(m,2H),2.66(s,3H),1.47(d,J＝6.8Hz,3H).
实施例39：N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1-(甲基磺酰基)-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a22-29)的制备
制备方法同实施例38，不同之处在于将乙酰氯替换为甲磺酰氯。
1H NMR(400MHz,DMSO-d 6)δ8.54(d,J＝7.6Hz,1H),8.37(s,1H),8.32(s,1H),8.26(s,1H),8.15(d,J＝7.2Hz,1H),7.11–6.99(m,3H),6.44(d,J＝7.6Hz,1H),5.61–5.51(m,1H),4.97–4.89(m,1H),4.86–4.75(m,1H),4.53–4.29(m,2H),3.51(s,3H),1.48(d,J＝6.8Hz,3H).。
实施例40：(R)-N-(1-(5-氟-2-(2,2,2-三氟乙氧基)苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a23-31)的制备
采用与实施例21相似的方法，所不同的是，将二氟一碘甲烷替换为2-碘-1,1,1-三氟乙烷。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.02(s,1H),8.00(d,J＝6.8Hz,1H),7.70(d,J＝2.4Hz,2H),7.25–7.01(m,3H),6.35(d,J＝7.6Hz,1H),5.53–5.41(m,1H),4.99–4.81(m,2H),3.80(s,3H),1.44(d,J＝6.4Hz,3H).
实施例41：(R)-2-(4-(5-((1-(5-氟-2-(2,2,2-三氟乙氧基)苯基)乙基)氨基)吡唑并[1,5-a]嘧啶-3-基)-1H-吡唑-1-基)乙醇(化合物a24-32)的制备
采用与实施例21相似的方法，所不同的是，将碘乙烷替换为2-碘-1,1,1-三氟乙烷，以及将1-甲基吡唑-4-硼酸频哪醇酯替换为4-(4,4,5,5-四甲基-1,3,2-二噁硼烷-2-基)-1H-吡唑-1-乙醇。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.2Hz,1H),8.03(s,1H),7.98(d,J＝6.4Hz,1H),7.73(d,J＝11.2Hz,2H),7.27–7.00(m,3H),6.35(d,J＝7.6Hz,1H),5.51–5.41(m,1H),5.00–4.78(m,3H),4.08(t,J＝5.6Hz,2H),3.71(q,J＝5.6Hz,2H),1.44(d,J＝6.8Hz,3H).
实施例42：N-(1-(5-氟-2-(3-氟代丙氧基)苯基)乙基)-3-(1-(四氢-2H-吡喃-4-基)-1H-吡唑-4-基)吡唑并[1，5-a]嘧啶-5-胺(化合物a25-35)的制备
采用与实施例24相似的方法，所不同的是，将碘乙烷替换为3-氟-1-碘代丙烷。
1H NMR(400MHz,DMSO-d6)δ8.46(d,J＝7.6Hz,1H),8.03(s,1H),7.93(d,J＝7.2Hz,1H),7.84(s,1H),7.79(s,1H),7.12–6.94(m,3H),6.36(d,J＝7.6Hz,1H),5.49(t,J＝7.2Hz,1H),4.73(td,J＝6.0,2.4Hz,1H),4.61(td,J＝6.0,2.4Hz,1H),4.40–4.07(m,3H),3.99(dq,J＝11.2,3.2Hz,2H),3.49(tt,J＝11.2,3.2Hz,2H),2.22(dp,J＝24.4,6.0Hz,2H),1.92(td,J＝11.2,10.4,4.4Hz,4H),1.45(d,J＝6.8Hz,3H).
实施例43：N-(1-(2-(环丙基甲氧基)-5-氟苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a26-38)的制备
采用与实施例24相似的方法，所不同的是，将碘乙烷替换为碘甲基环丙烷，以及将1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯替换为1-甲基吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.48(d,J＝7.6Hz,1H),8.03(s,1H),7.99(d,J＝6.8Hz,1H),7.76(d,J＝2.8Hz,2H),7.14–6.90(m,3H),6.37(d,J＝7.6Hz,1H),5.58–5.49(m,1H),4.09–3.93(m,2H),3.84(s,3H),1.47(d,J＝6.8Hz,3H),1.39–1.29(m,1H),0.64–0.56(m,2H),0.46–0.39(m,2H).
实施例44：N-(1-(5-氟-2-((四氢呋喃-3-基)氧基)苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物a27-41)的制备
采用与实施例24相似的方法，所不同的是，将碘乙烷替换为3-碘四氢呋喃，以及将1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯替换为1-甲基吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.03(s,1H),7.94(d,J＝7.0Hz,1H),7.78(s,1H),7.75(s,1H),7.16–6.94(m,3H),6.37(d,J＝7.6Hz,1H),5.51–5.43(m,1H),5.21–5.15(m,1H),4.10–3.76(m,7H),2.40–1.96(m,2H),1.44(d,J＝6.8Hz,3H).
实施例45：N-(1-(5-氟-2-((四氢-2H-吡喃-4-基)甲氧基)苯基)乙基)-3-(1-甲基-1H-吡唑-4-基)吡唑并[1,25-a]嘧啶-5-胺(化合物a28-43)的制备
采用与实施例24相似的方法，所不同的是，将碘乙烷替换为4-碘甲基四氢吡喃，以及将1-(四氢吡喃-4-基)-1H-吡唑-4-硼酸频哪醇酯替换为1-甲基吡唑-4-硼酸频哪醇酯。
1H NMR(400MHz,DMSO-d 6)δ8.47(d,J＝7.6Hz,1H),8.02(s,1H),7.95(d,J＝6.8Hz,1H),7.75(s,2H),7.14–6.93(m,3H),6.36(d,J＝7.6Hz,1H),5.58–5.48(m,1H),4.07–3.98(m,1H),3.97–3.77(m,6H),3.31–3.28(m,2H),2.18–2.06(m,1H),1.83–1.67(m,2H),1.55–1.35(m,5H).
实施例46：2-(1-((3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-基)氨基)乙基)-4-氟苯酚(化合物b1-1)的制备
Figure PCTCN2020080198-appb-000079
步骤1)：将1-(5-氟-2-羟基苯基)乙酮(32.4mmol)，4-甲氧基溴苄(38.9mmol)，碳酸铯(48.6mmol)加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。油浴(80℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率78％。
步骤2)：将1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙酮(25.3mmol)，盐酸羟胺(30.4mmol)，碳酸铯(38.0mmol)加入到200mL的梨形瓶中，向其中加入甲醇(50mL)。在室温，磁力搅拌条件下，反应4h。将反应液倒入水中，抽滤，干燥，得到白色固体，收率98％。
步骤3)：将1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙酮肟(24.8mmol)，锌粉(372mmol)，氯化铵(372mmol)，乙酸(372mmol)加入到200mL的梨形瓶中，向其中加入甲醇(50mL)。油浴(80℃)中加热过夜。将反应冷却至环境温度，中和，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黄色液体，收率77％。
步骤4)：将5-氯吡唑并[1,5-a]嘧啶(19.1mmol)，1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙胺(19.1mmol)，无水正丁醇(50mL)和N，N-二异丙基乙胺(DIPEA，38.2mmol)加入到200mL的梨形瓶中。在油浴(140℃)中加热过夜。将反应冷却至环境温度，减压浓缩尽量除去正丁醇和N，N-二异丙基乙胺(DIPEA)得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率68％。
步骤5)：将N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(13.0mmol)，加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS， 14.3mmol)。室温反应1h，TLC监测反应完毕。减压尽量出去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率67％。
步骤6)：将N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.77mmol)，1-Boc-吡唑-4-硼酸频哪醇酯(1.16mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.08mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2mL水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率50％。
步骤7)：将N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(0.22mmol)，加入到200mL的梨形瓶中，向其中加入二氯甲烷(10mL)，滴加三氟乙酸(3.3mmol)。在室温，磁力搅拌条件下，反应4h。中和并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，柱层析纯化得白色固体，收率85％。
1H NMR(400MHz,DMSO-d 6)δ12.63(s,1H),9.66–9.56(m,1H),8.44(d,J＝7.6Hz,1H),8.05(s,1H),7.97–7.82(m,3H),7.08–6.98(m,1H),6.89–6.76(m,2H),6.32(d,J＝7.6Hz,1H),5.57–5.46(m,1H),1.43(d,J＝6.8Hz,3H).
实施例47：N-(1-(5-氟-2-甲氧基苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b2-5)的制备
Figure PCTCN2020080198-appb-000080
步骤1)：将N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(13.0mmol)，加入到200mL的梨形瓶中，向其中加入二氯甲烷(50mL)，滴加三氟乙酸(195mmol)。在室温，磁力搅拌条件下，反应4h。中和并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，柱层析纯化得白色固体，收率85％。
步骤2)：将4-氟-2-(1-(吡唑并[1,5-a]嘧啶-5-基氨基)乙基)苯酚(11.0mmol)，碘甲烷(16.5mmol)，碳酸铯(22mmol)加入到200mL的梨形瓶中，向其中加入DMF(50mL)。油浴(100℃)中加热过夜。将反应冷却至环境温度，粗品经柱层析纯化得白色固体，收率90％。
步骤3)：将N-(1-(5-氟-2-甲氧基苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(9.8mmol)，加入到200mL的梨形瓶中，向其中加入乙腈(50mL)。在室温，磁力搅拌条件下，加入N-碘代丁二酰亚胺(NIS，14.85mmol)。室温反应1h，TLC监测反应完毕。减压尽量出去乙腈后用250mL乙酸乙酯稀释，并转移到分液漏斗中。用1mol/L得NaOH洗3次，饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得红色油状粗品，粗品经柱层析纯化得淡黄色固体，收率67％。
步骤4)：将N-(1-(5-氟-2-甲氧基苯基)乙基)-3-碘吡唑并[1,5-a]嘧啶-5-胺(0.50mmol)，1-Boc-吡唑-4-硼酸频哪醇酯(0.75mmol)，无水碳酸钾(2.00mmol)，四(三苯基膦)钯(0.05mmol)加入到100mL反应管中，氩气置换3次，加入10mL无水DMF,2ml水。100℃，氩气氛围下反应2h，TLC监测反应完毕。冷却至50℃，硅藻土过滤，滤液加水并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，浓缩得黑色油状粗品，粗品经柱层析纯化得淡黄色固体，收率50％。
1H NMR(400MHz,DMSO-d6)δ12.70(s,1H),8.44(d,J＝7.6Hz,1H),8.14–7.91(m,2H),7.82(s,2H),7.16–6.87(m,3H),6.32(d,J＝8.0Hz,1H),5.57–5.40(m,1H),3.95(s,3H),1.40(d,J＝7.2Hz,3H).
实施例48：N-(1-(5-氟-2-(氟甲氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b3-6)的制备
Figure PCTCN2020080198-appb-000081
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为氟代碘甲烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法用实施例47中的步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.68(s,1H),8.46(d,J＝7.6Hz,1H),8.04(d,J＝3.2Hz,1H),8.02(s,1H),7.86(s,1H),7.79(s,1H),7.27–7.02(m,3H),6.33(d,J＝7.6Hz,1H),6.07(s,1H),5.93(s,1H),5.58–5.47(m,1H),1.45(d,J＝6.8Hz,3H).
实施例49：N-(1-(2-(二氟甲氧基)-5-氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b4-7)的制备
Figure PCTCN2020080198-appb-000082
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为二氟一碘甲烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法用实施例47中的步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.70(s,1H),8.50(d,J＝7.6Hz,1H),8.08(d,J＝1.6Hz,1H),8.03(d,J＝6.8Hz,1H),7.85(s,2H),7.52–7.07(m,4H),6.36(d,J＝7.6Hz,1H),5.50(t,J＝7.2Hz,1H),1.50(d,J＝6.8Hz,3H).
实施例50：N-(1-(2-乙氧基-5-氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b5-11)的制备
Figure PCTCN2020080198-appb-000083
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为碘乙烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法用实施例47中的步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.70(s,1H),8.47(d,J＝7.2Hz,1H),8.06(s,1H),8.00(d,J＝7.2Hz,1H),7.87(s,2H),7.09(dd,J＝9.6,3.0Hz,1H),7.04–6.91(m,2H),6.35(d,J＝7.2Hz,1H),5.65–5.53(m,1H),4.27–4.10(m,2H),1.51–1.41(m,6H).
实施例51：N-(1-(5-氟-2-(2-甲氧基乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b6-15)的制备
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为2-碘乙基甲醚。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法用实施例47中的步骤4。
1H NMR(400MHz,DMSO-d6)δ12.67(s,1H),8.45(d,J＝7.6Hz,1H),8.05(s,1H),7.94(d,J＝7.2Hz,1H),7.86(s,2H),7.16–6.87(m,3H),6.34(d,J＝7.6Hz,1H),5.63–5.46(m,1H),4.33–4.12(m,2H),3.85–3.68(m,2H),3.44–3.38(m,3H),1.44(d,J＝6.8Hz,3H).
实施例52：(R)-N-(1-(5-氟-2-(2-氟乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b7-16)的制备
Figure PCTCN2020080198-appb-000084
步骤1)：将5-氯吡唑并[1,5-a]嘧啶(19.1mmol)，(R)-1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙胺(19.1mmol)，无水正丁醇(50mL)和N，N-二异丙基乙胺(DIPEA，38.2mmol)加入到200mL的梨形瓶中。在油浴(140℃)中加热过夜。将反应冷却至环境温度，减压浓缩尽量除去正丁醇和N，N-二异丙基乙胺(DIPEA)得黄色油状粗品，粗品经柱层析纯化得淡黄色固体，收率68％
步骤2)：将(R)-N-(1-(5-氟-2-((4-甲氧基苄基)氧基)苯基)乙基)吡唑并[1,5-a]嘧啶-5-胺(13.0mmol)，加入到200mL的梨形瓶中，向其中加入二氯甲烷(50mL)，滴加三氟乙酸(195mmol)。在室温，磁力搅拌条件下，反应4h。中和并用乙酸乙酯萃取。有机相用饱和食盐水洗两次，用无水硫酸钠干燥，柱层析纯化得白色固体，收率85％。
步骤3)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为1-氟-2-碘乙烷。
步骤4)：采用实施例47中的步骤3的方法。
步骤5)：合成方法同实施例47中步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.68(s,1H),8.48(d,J＝7.2Hz,1H),8.07(s,1H),8.02(d,J＝7.2Hz,1H),7.86(s,2H),7.16–6.95(m,3H),6.36(d,J＝7.6Hz,1H),5.64–5.54(m,1H),4.94(t,J＝3.6Hz,1H),4.82(t,J＝3.6Hz,1H),4.48–4.44(m,1H),4.41–4.36(m,1H),1.46(d,J＝7.2Hz,3H).
实施例53：(R)-N-(1-(5-氟-2-(2,2,2-三氟乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b8-30)的制备
Figure PCTCN2020080198-appb-000085
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为2-碘-1,1,1-三氟乙烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.67(s,1H),8.46(d,J＝7.6Hz,1H),8.04(s,1H),8.02(d,J＝7.2Hz,1H),7.80(s,2H),7.08(m,3H),6.34(d,J＝7.6Hz,1H),5.52–5.44(m,1H),5.04–4.70(m,2H),1.43(d,J＝6.4Hz,3H).
实施例54：N-(1-(5-氟-2-丙氧基苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b9-33)的制备
Figure PCTCN2020080198-appb-000086
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为碘丙烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.67(s,1H),8.45(d,J＝7.6Hz,1H),8.04(s,1H),7.98(d,J＝7.2Hz,1H),7.83(s,2H),7.10–6.90(m,3H),6.33(d,J＝7.6Hz,1H),5.61–5.48(m,1H),4.13(dt,J＝9.2,6.0Hz,1H),4.03(dt,J＝9.2,6.4Hz,1H),1.84(q,J＝6.8Hz,2H),1.42(d,J＝6.8Hz,3H),1.05(t,J＝7.6Hz,3H).
实施例55：N-(1-(5-氟-2-(3-氟代丙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b10-34)的制备
Figure PCTCN2020080198-appb-000087
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为3-氟-1-碘代丙烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.70(s,1H),8.46(d,J＝7.6Hz,1H),8.06(s,1H),7.98(d,J＝7.2Hz,1H),7.86(s,2H),7.15–6.90(m,3H),6.35(d,J＝7.6Hz,1H),5.60–5.46(m,1H),4.74(tt,J＝5.6,2.8Hz,1H),4.62(td,J＝6.0,2.4Hz,1H),4.29–4.15(m,2H),2.30–2.17(m,2H),1.44(d,J＝6.8Hz,3H).
实施例56：2-(2-(1-((3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-基)氨基)乙基)-4-氟苯氧基)乙腈(化合物b11-36)的制备
Figure PCTCN2020080198-appb-000088
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为碘乙睛。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法用实施例47中的步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.64(s,1H),8.48(d,J＝7.2Hz,1H),8.06(s,1H),7.86(s,1H),7.50(s,1H),7.41(s,1H),7.12(d,J＝9.0Hz,1H),7.09–6.92(m,2H),6.36(d,J＝7.2Hz,1H),5.66–5.49(m,1H),4.76–4.50(m,2H),1.51(d,J＝6.6Hz,3H).
实施例57：N-(1-(2-(环丙基甲氧基)-5-氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b12-37)的制备
Figure PCTCN2020080198-appb-000089
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为(碘甲基)环丙烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.68(s,1H),8.47(d,J＝7.6Hz,1H),8.06(s,1H),7.98(d,J＝7.2Hz,1H),7.85(s,2H),7.12–6.91(m,3H),6.36(d,J＝7.6Hz,1H),5.66–5.50(m,1H),4.01(qd,J＝10.0,6.8Hz,2H),1.46(d,J＝6.8Hz,3H),1.40–1.31(m,1H),0.65–0.54(m,2H),0.47–0.41(m,2H).
实施例58：N-(1-(2-(环戊氧基)-5-氟苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b13-39)的制备
Figure PCTCN2020080198-appb-000090
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为碘代环戊烷。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(600MHz,DMSO-d 6)δ12.66(s,1H),8.45(d,J＝7.2Hz,1H),8.05(s,1H),7.91(s,1H),7.85(s,2H),7.05–6.96(m,3H),6.37–6.28(m,1H),5.55–5.48(m,1H),4.99–4.89(m,1H),2.05–1.55(m,8H),1.40(d,J＝6.6Hz,3H).
实施例59：N-(1-(5-氟-2-((四氢呋喃-3-基)氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b14-40)的制备
Figure PCTCN2020080198-appb-000091
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为3-碘四氢呋喃。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.68(s,1H),8.47(d,J＝7.6Hz,1H),8.06(s,1H),7.96(t,J＝6.0Hz,1H),7.85(s,1H),7.84(s,1H),7.17–6.95(m,3H),6.36(d,J＝7.6Hz,1H),5.53–5.46(m,1H),5.21–5.16(m,1H),4.14–3.76(m,4H),2.37–2.00(m,2H),1.43(d,J＝6.8Hz,3H).
实施例60：N-(1-(5-氟-2-((四氢-2H-吡喃-4-基)甲氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b15-42)的制备
Figure PCTCN2020080198-appb-000092
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为4-碘甲基四氢吡喃。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(400MHz,DMSO-d 6)δ12.65(s,1H),8.47(d,J＝7.6Hz,1H),8.06(s,1H),7.98(d,J＝6.8Hz,1H),7.84(s,2H),7.13–6.94(m,3H),6.35(d,J＝7.6Hz,1H),5.60–5.51(m,1H),4.11–3.99(m,1H),3.99–3.79(m,3H),3.32–3.30(m,2H),2.20–2.05(m,1H),1.77(t,J＝12.4Hz,2H),1.50–1.41(m,5H).
实施例61：N-(1-(5-氟-2-(2-吗啉代乙氧基)苯基)乙基)-3-(1H-吡唑-4-基)吡唑并[1,5-a]嘧啶-5-胺(化合物b16-44)的制备
Figure PCTCN2020080198-appb-000093
步骤1)：采用实施例47中的步骤2的方法，不同之处在于将碘甲烷替换为2-(4-吗啉)乙基溴。
步骤2)：采用实施例47中的步骤3的方法。
步骤3)：合成方法同实施例47中步骤4。
1H NMR(600MHz,DMSO-d 6)δ12.68(s,1H),8.46(d,J＝7.8Hz,1H),8.04(s,1H),8.00–7.90(m,1H),7.83(s,2H),7.13–6.91(m,3H),6.33(d,J＝7.8Hz,1H),5.60–5.45(m,1H),4.32–4.13(m,2H),3.59–3.44(m,4H),2.81(t,J＝6.0Hz,2H),2.56–2.47(m,4H),1.43(d,J＝6.6Hz,3H).
测试例1：体外生化水平抑制蛋白激酶(PK)活性实验
材料与方法：TRKA、TRKB和TRKC等野生型激酶和TRKA-G595R、TRKA-G667C、TRKA-F589L、TRKC-G623R、TRKC-G696A等突变的激酶，来源于Carna Biosciences 08-186，08-187，08-197；HTRF KinEASE TK kit(Cisbio公司62TK0PEC)；384孔板(Greiner公司)；ATP(Life technologies PV3227)，MgCl 2(sigma) 公司；PHERAstar FS多功能酶标仪(BMG公司)；低速离心机(StaiteXiangyi公司)；恒温箱(Binder公司)。
选取的对照化合物为WO2007147647中公开的典型化合物A、WO2007025540中公开的典型化合物B，以及对比化合物C、对比化合物D，其中，对比化合物C(核磁数据为： 1H NMR(400MHz,DMSO-d 6)δ12.78–12.66(m,1H),8.45(d,J＝7.6Hz,1H),8.05(s,1H),8.01–7.80(m,3H),6.94(s,1H),6.90(d,J＝9.2Hz,1H),6.77(dt,J＝10.8,2.4Hz,1H),6.56–6.21(m,2H),5.20(d,J＝6.0Hz,1H),4.31(tt,J＝14.4,3.6Hz,2H),1.48(d,J＝6.8Hz,3H))和对比化合物D(核磁数据为： 1H NMR(400MHz,DMSO-d 6)δ12.70(br,1H),8.44(d,J＝7.6Hz,1H),8.05(s,1H),7.94(d,J＝6.8Hz,2H),7.85(s,1H),7.35–7.14(m,3H),6.54–6.20(m,2H),5.28–5.11(m,1H),4.39–4.30(m,2H),1.48(d,J＝6.8Hz,3H))的制备方法参照前述实施例1的制备方法，仅是原料不同，结构如下：
Figure PCTCN2020080198-appb-000094
化合物溶解及保存：根据溶解性，用二甲基亚砜(DMSO)将受试化合物配置成10mmol/L的母液，分装后-20℃保存；
化合物工作液的配制：测试前将分装的化合物从冰箱取出，用纯DMSO稀释到100×所需浓度；然后用去离子水将化合物稀释至4×所需浓度；
1.33×酶缓冲液(Enzymatic buffer)的配制：将5×酶缓冲液(来源于HTRF试剂盒)用去离子水稀释到1.33×，并且加入1.33×终浓度的相应成分：1.33mmol/L二硫苏糖醇(DTT)、1.33mmol/L MnCl 2、6.65mmol/L MgCl 2和39.9nmol/L SEB；
激酶工作液的配制：用1.33×酶缓冲液将TRKA、TRKB和TRKC稀释到2×所需浓度分别为0.404ng/μL、0.304ng/μL和0.236ng/μL；
底物工作液的配制：用1.33×酶缓冲液将TK Substrate Blr稀释至物(来源于HTRF试剂盒)和ATP(10mM)稀释为4×所需终浓度的混合液；TRKA、TRKB和TRKC的ATP终浓度分别为：3.727μmol/L、2.56μmol/L和2.526μmol/L。TK Substrate–biotin(来自HTRF KinEASE TK kit)终浓度均为：0.2μmol/L。
检测工作液的配制：用HTRF测试缓冲液将16.67μmol/L的抗生蛋白链菌-XL665(Streptavidin-XL665)稀释到4×所需终浓度，然后与等体积的抗体-铕隐酸盐(Antibody-Cryptate)混合(均来源于HTRF试剂盒)。
酶反应步骤：向低体积384微孔板的每个孔中加入4μL的激酶工作液，同时加入4μL的1.33×酶缓冲液作为阴性对照(Negative)；向孔加入2μl的化合物工作液，同时加入2μL的8％DMSO水溶液作为零化合物浓度对照(即阳性对照，Positive)；于25℃，孵育5min；向孔中加入2μL底物工作液启动酶反应，于37℃，振荡反应30min。
HTRF试剂检测步骤：向孔加入8检测的检测工作液终止反应；25℃反应1h；
HTRF信号的读取：采用PHERAstar FS读数检测信号，仪器相应设置如下：
Optic module
Figure PCTCN2020080198-appb-000095
积分延迟(Integration delay，lag time)50μs
积分时间(Integration time)400μs
闪光次数(Number of flashes)200
对于每孔读出的原始数据，比值＝665nm/620nm；
抑制率的计算：
Figure PCTCN2020080198-appb-000096
IC 50值的计算：以化合物浓度的对数为横坐标，抑制率为纵坐标，在GraphPad Prism 5中，拟合非线性曲线：log(inhibitor)vs.response--Variable slope，求出酶活抑制率为50％时的待测化合物浓度即IC 50。
实验结果：TRKA、TRKB和TRKC激酶活性半数抑制浓度(IC 50,nM)
本发明提供结构如式(I)所示化合物以及对照化合物对TRKA、TRKB和TRKC的半数抑制浓度(IC 50)见 表1：
表1：化合物的TRKA、TRKB和TRKC激酶抑制活性
Figure PCTCN2020080198-appb-000097
Figure PCTCN2020080198-appb-000098
如表1所示，本发明提供的化合物对野生的TRKA、TRKB和TRKC激酶均表现出优异的抑制活性，显著优于典型化合物A、典型化合物B、对比化合物C和对比化合物D。
测试例2：大鼠体内药物代谢研究
将前述部分实施例提供的化合物以聚乙二醇400水溶液(70％)形式大鼠给药。对于口服给药，大鼠给予5mg/kg的剂量。口服组给药后15、30、45min、1、2、4、6、8、10、24h各采集血样约0.3mL至肝素化的Eppendorf管中，于冰上暂存至离心。全血经8000rpm离心5min后收集血浆，转移血浆至96孔板中，于-20℃保存至LC-MS/MS检测。
采用软件WinNonlin软件的非室模型计算大鼠给药后的药代动力学参数。
达峰浓度C max：采用实测值；
药时曲线下面积AUC0-t值：采用梯形法计算；AUC 0-∞＝AUC 0-t+Ct/ke，Ct为最后一个可测得时间点的血药浓度，ke为消除速率常数；
消除半衰期t 1/2＝0.693/ke；
绝度生物利用度F＝Dose iv*AUC 0-t，ig/Dose ig*AUC 0-t，iv×100％。
表2列出了静注给药后，本发明的化合物在大鼠体内的药代动力学参数。结果表明，本发明的化合物具有良好的药代动力学性质，包括理想的清除率(CL)、半衰期(t 1/2)、峰浓度(C max)和暴露量(AUC 0-t)。
表3列出了口服给药后，本发明的化合物在大鼠体内的药代动力学参数。结果表明，本发明的化合物具有良好的药代动力学性质，包括理想的清除率(CL)、半衰期(t 1/2)、峰浓度(C max)、暴露量(AUC 0-t)和口服生物利用度。
表2：化合物的大鼠静注给药的主要药代动力学参数
Figure PCTCN2020080198-appb-000099
Figure PCTCN2020080198-appb-000100
表3：化合物的大鼠口服给药的主要药代动力学参数
Figure PCTCN2020080198-appb-000101
测试例3：本发明的化合物对五种TRK激酶突变体的抑制活性
本测试例采用与测试例1相同的测试方法进行。
本测试例的结果列于表4中。
表4
Figure PCTCN2020080198-appb-000102
由表4可见，本发明的化合物对五种TRK激酶突变体的抑制活性优于对野生的TRK激酶的活性，有望有效克服临床报道的肿瘤耐药性。
测试例4：本发明的化合物在裸鼠异体移植瘤模型上的抗肿瘤活性
本发明化合物的药效通过移植肿瘤的标准鼠类模型来进行评价。人非小细胞肺癌H2228培养、收集后，于后腹侧皮下接种于5-6周龄的雌性裸小鼠体内(BALB/c，上海灵畅生物科技有限公司)。当肿瘤体积达到100-150mm 3时，动物随机地分为溶剂对照组(70％PEG-400的水溶液)和化合物组(每组6只动物)。后续采用实施例的化合物对动物进行灌胃给药(相应剂量，溶解在70％PEG-400的水溶液中)，从肿瘤细胞接种后的0到7天中的任何地方开始，并且在试验中每天进行两次。
实验指标为考察实施例化合物对肿瘤生长的影响，具体指标为T/C％或抑瘤率TGI(％)。
每周二次用游标卡尺测量肿瘤直径，肿瘤体积(V)计算公式为：
V＝1/2×a×b 2其中a、b分别表示长、宽。
T/C(％)＝(T-T 0)/(C-C 0)×100其中T、C为实验结束时的肿瘤体积；T 0、C 0为实验开始时的肿瘤体积。
抑瘤率(TGI)(％)＝100-T/C(％)。
当肿瘤出现消退时，抑瘤率(TGI)(％)＝100-(T-T 0)/T 0×100
如果肿瘤比起始体积缩小，即T<T 0或C<C 0时，即定义为肿瘤部分消退(PR)；如果肿瘤完全消失，即定义为肿瘤完全消退(CR)。
二组肿瘤体积之间比较采用双尾Student’s t检验，P<0.05定义为有统计学显著性差异。
本测试例的结果列于表5中。
以下BID表示一天两次给药。
表5
分组 给药 抑瘤率D14 完全消退 部分消退
溶剂 BID,D0-7 - - -
化合物5,50mg/kg BID,D0-7 155％ 3/6 2/6
由表5可见，本发明的化合物在人非小细胞肺癌H2228裸鼠异体移植瘤模型上表现出优异的抗肿瘤活性。其中，化合物5(50mg/kg,BID×8)明显抑制人非小细胞肺癌H2228裸小鼠皮下移植瘤的生长，给药至D7时，抑瘤率为182％，4/6肿瘤完全消退；从D8开始停止给药，至实验结束(D14)，抑瘤率为155％，3/6动物肿瘤完全消退，2/6动物肿瘤出现部分消退。
测试例5：本发明的化合物在裸鼠异体移植瘤模型上的抗肿瘤活性
本发明化合物的药效通过移植肿瘤的标准鼠类模型来进行评的。人非小细胞肺癌H2228培养、收集后，于后腹侧皮下接种于5-6周龄的雌性裸小鼠体内(BALB/c，上海灵畅生物科技有限公司)。当肿瘤体积达到100-150mm 3时，动物随机地分为溶剂对照组(70％PEG-400的水溶液)和化合物组(每组6只动物)。后续采用实施例的化合物对动物进行灌胃给药(相应剂量，溶解在70％PEG-400的水溶液中)，从肿瘤细胞接种后的0到13天中的任何地方开始，并且在试验中每天进行一次或两次。
实验指标为考察实施例化合物对肿瘤生长的影响，具体指标为T/C％或抑瘤率TGI(％)。
每周二次用游标卡尺测量肿瘤直径，肿瘤体积(V)计算公式为：
V＝1/2×a×b 2其中a、b分别表示长、宽。
T/C(％)＝(T-T 0)/(C-C 0)×100其中T、C为实验结束时的肿瘤体积；T 0、C 0为实验开始时的肿瘤体积。
抑瘤率(TGI)(％)＝100-T/C(％)。
当肿瘤出现消退时，抑瘤率(TGI)(％)＝100-(T-T 0)/T 0×100
如果肿瘤比起始体积缩小，即T<T 0或C<C 0时，即定义为肿瘤部分消退(PR)；如果肿瘤完全消失，即定义为肿瘤完全消退(CR)。
二组肿瘤体积之间比较采用双尾Student’s t检验，P<0.05定义为有统计学显著性差异。
本测试例的结果列于表6中。
以下BID表示一天两次给药，QD表示一天一次给药。
表6
分组 给药 抑瘤率D14 部分消退
溶剂 BID,D0-7 - -
化合物b8-30 25mg/kg BID,D0-13 127％ 6/6
化合物b8-30 50mg/kg QD,D0-10 110％ 3/6
由表6可见，本发明的化合物在人非小细胞肺癌H2228裸鼠异体移植瘤模型上表现出优异的抗肿瘤活性。其中，化合物b8-30(25mg/kg,BID×14)明显抑制人非小细胞肺癌H2228裸小鼠皮下移植瘤的生长，给药至D7时，抑瘤率为95％，从D8开始增加剂量至50mg/kg，肿瘤出现消退，至实验结束(D14)，抑瘤率为127％，6/6肿瘤出现部分消退；化合物b8-30(50mg/kg,QD,QD×11)对H2228裸小鼠皮下移植瘤的抑瘤率为96％(D7)，从D8开始增加剂量至100mg/kg，肿瘤出现消退，至实验结束(D14)，抑瘤率为110％，3/6肿瘤出现部分消退。
上述结果表明，本发明提供的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药对TRK激酶表现出优异的抑制活性，同时，能够在动物水平上表现出良好的抗肿瘤活性。
以上详细描述了本发明的优选实施方式，但是，本发明并不限于此。在本发明的技术构思范围内，可以对本发明的技术方案进行多种简单变型，包括各个技术特征以任何其它的合适方式进行组合，这些简单变型和组合同样应当视为本发明所公开的内容，均属于本发明的保护范围。

Claims (15)

  1. 一种具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药，
    Figure PCTCN2020080198-appb-100001
    其中，在式(I)中，
    R 1、R 2、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
    R 5选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、烷氧基取代的C 2-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基；
    R 6选自H、C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；
    R 7选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基。
  2. 根据权利要求1所述的化合物，其中，在式(I)中，
    R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
    R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
    R 6选自H、C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
    R 7选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基；
    优选地，在式(I)中，
    R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基；
    R 5选自H、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基；
    R 6选自H、C 1-6的烷基、羟基取代的C 1-6的烷基和卤素；
    R 7选自H、C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基、C 2-6的酰基、磺酰基。
  3. 根据权利要求1或2所述的化合物，其中，在式(I)中，
    R 1、R 2、R 3、R 4和R 5中的至少一个基团中含有F原子；
    且R 5选自由1-6个卤素取代的C 1-12的烷基；
    优选地，在式(I)中，
    R 1、R 2、R 3、R 4和R 5中的至少一个基团中含有F原子；
    且R 5选自由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基。
  4. 根据权利要求3所述的化合物，其中，在式(I)中，
    R 1、R 2、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
    R 5为-CH 2CHF 2；
    R 6选自C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；
    R 7选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基；
    优选地，在式(I)中，
    R 1、R 2、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
    R 5为-CH 2CHF 2；
    R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
    R 7选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 1-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基；
    优选地，在式(I)中，
    R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
    R 2为H或F；
    R 5为-CH 2CHF 2；
    R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
    R 7选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基。
  5. 根据权利要求4所述的化合物，其中，式(I)所示结构的化合物选自以下化合物中的至少一种：
    Figure PCTCN2020080198-appb-100002
  6. 根据权利要求1或2所述的化合物，其中，在式(I)中，
    R 7选自C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基；
    优选地，在式(I)中，
    R 7选自C 1-6的烷基、由1-4个选自氟、氯和溴的卤素取代的C 1-6的烷基、羟基取代的C 1-6的烷基、氰基取代的C 2-6的烷基、含有1-3个选自N、O和S的杂原子的C 2-8的环烷基、C 2-6的酰基、磺酰基。
  7. 根据权利要求6所述的化合物，其中，在式(I)中，
    R 1、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
    R 2为卤素；
    R 5选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、烷氧基取代的C 2-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基；
    R 6选自C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；
    R 7选自C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基、C 2-12的酰基、磺酰基；
    优选地，在式(I)中，
    R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
    R 2为F；
    R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
    R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
    R 7选自C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 1-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基。
  8. 根据权利要求7所述的化合物，其中，在式(I)中，
    R 1、R 3和R 4均为H；R 2为F；
    R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
    R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
    R 7选自C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基、C 2-8的酰基、磺酰基；
    优选地，式(I)所示结构的化合物选自以下化合物中的至少一种：
    Figure PCTCN2020080198-appb-100003
    Figure PCTCN2020080198-appb-100004
  9. 根据权利要求1或2所述的化合物，其中，在式(I)中，R 7为H。
  10. 根据权利要求9所述的化合物，其中，在式(I)中，
    R 1、R 3和R 4各自独立地选自H、卤素、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基；
    R 2为卤素；
    R 5选自H、C 1-12的烷基、由1-6个卤素取代的C 1-12的烷基、羟基取代的C 1-12的烷基、烷氧基取代的C 2-12的烷基、氰基取代的C 2-12的烷基、含有1-3个选自N、O和S的杂原子的C 2-12的环烷基；
    R 6选自C 1-12的烷基、羟基取代的C 1-12的烷基和卤素；
    优选地，在式(I)中，
    R 1、R 3和R 4各自独立地选自H、氟、氯、溴、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基；
    R 2为F；
    R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
    R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素；
    优选地，在式(I)中，
    R 1、R 3和R 4均为H；R 2为F；
    R 5选自H、C 1-8的烷基、由1-6个选自氟、氯和溴的卤素取代的C 1-8的烷基、羟基取代的C 1-8的烷基、烷氧基取代的C 2-8的烷基、氰基取代的C 2-8的烷基、含有1-3个选自N、O和S的杂原子的C 2-10的环烷基；
    R 6选自C 1-8的烷基、羟基取代的C 1-8的烷基和卤素。
  11. 根据权利要求10所述的化合物，其中，式(I)所示结构的化合物选自以下化合物中的至少一种：
    Figure PCTCN2020080198-appb-100005
  12. 权利要求1-11中任意一项所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药在制备用于预防和/或治疗TRK激酶介导的疾病的药物中的应用。
  13. 一种药物组合物，其包含药学上可接受的载体、赋形剂或稀释剂，以及作为活性成分的权利要求1-11中任意一项所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药。
  14. 权利要求13所述的药物组合物在制备用于预防和/或治疗TRK激酶受体介导的疾病的药物中的应用。
  15. 权利要求1-11中任意一项所述的具有式(I)所示结构的吡唑并嘧啶化合物或其药学上可接受的盐，或其立体异构体、几何异构体、互变异构体、氮氧化物、水合物、溶剂化物、代谢产物，或其前药，或者权利要求13所述的药物组合物在制备用于预防和/或治疗肿瘤的药物中的应用；
    优选地，所述肿瘤为乳腺癌、大肠癌、肺癌、甲状腺癌、皮肤癌、骨癌、黑色素瘤、白血病、唾液腺肿瘤、神经内分泌肿瘤、淋巴瘤、脑肿瘤、成神经细胞瘤、卵巢癌、胰腺癌、间皮瘤、食管癌、肺肉瘤、成神经管细胞瘤、成胶质细胞瘤、结肠癌、肝癌、成视网膜细胞瘤、肾癌、膀胱癌、骨肉瘤、胃癌、子宫癌、外阴癌、小肠癌、前列腺癌、胆管癌、输尿管癌、肾上腺皮质癌或头颈部癌症的至少一种。
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Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007013673A1 (en) * 2005-07-29 2007-02-01 Astellas Pharma Inc. Fused heterocycles as lck inhibitors
WO2007025540A2 (de) 2005-09-02 2007-03-08 Bayer Schering Pharma Aktiengesellschaft Substituierte imidazo[1,2b]pyridazine als kinase-inhibitoren, deren herstellung und verwendung als arzneimittel
WO2007147647A1 (en) 2006-06-21 2007-12-27 Bayer Schering Pharma Aktiengesellschaft Pyrazolopyrimidines and salts thereof, pharmaceutical compositions comprising same, methods of preparing same and uses of same.
WO2008025822A1 (en) * 2006-08-30 2008-03-06 Cellzome Limited Diazolodiazine derivatives as kinase inhibitors
WO2010033941A1 (en) 2008-09-22 2010-03-25 Array Biopharma Inc. Substituted imidazo[1,2b]pyridazine compounds as trk kinase inhibitors
WO2010048314A1 (en) 2008-10-22 2010-04-29 Array Biopharma Inc. SUBSTITUTED PYRAZOLO[1,5-a]PYRIMIDINE COMPOUNDS AS TRK KINASE INHIBITORS
WO2011146336A1 (en) 2010-05-20 2011-11-24 Array Biopharma Inc. Macrocyclic compounds as trk kinase inhibitors
WO2012116217A1 (en) 2011-02-25 2012-08-30 Irm Llc Compounds and compositions as trk inhibitors
WO2018077246A1 (zh) 2016-10-28 2018-05-03 正大天晴药业集团股份有限公司 用作神经营养因子酪氨酸激酶受体抑制剂的氨基吡唑并嘧啶化合物
WO2019118584A1 (en) * 2017-12-15 2019-06-20 Pyramid Biosciences, Inc. 5-(2-(2,5-difluorophenyl)pyrrolidin-1 -yl)-3-(1h-pyrazol-1-yl)pyrazolo[1,5-a]pyrimidine derivatives and related compounds as trk kinase inhibitors for treating cancer

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107207514B (zh) * 2014-12-15 2020-01-24 康联制药有限公司 稠环杂芳基化合物及其作为trk抑制剂的用途

Patent Citations (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007013673A1 (en) * 2005-07-29 2007-02-01 Astellas Pharma Inc. Fused heterocycles as lck inhibitors
WO2007025540A2 (de) 2005-09-02 2007-03-08 Bayer Schering Pharma Aktiengesellschaft Substituierte imidazo[1,2b]pyridazine als kinase-inhibitoren, deren herstellung und verwendung als arzneimittel
WO2007147647A1 (en) 2006-06-21 2007-12-27 Bayer Schering Pharma Aktiengesellschaft Pyrazolopyrimidines and salts thereof, pharmaceutical compositions comprising same, methods of preparing same and uses of same.
WO2008025822A1 (en) * 2006-08-30 2008-03-06 Cellzome Limited Diazolodiazine derivatives as kinase inhibitors
WO2010033941A1 (en) 2008-09-22 2010-03-25 Array Biopharma Inc. Substituted imidazo[1,2b]pyridazine compounds as trk kinase inhibitors
WO2010048314A1 (en) 2008-10-22 2010-04-29 Array Biopharma Inc. SUBSTITUTED PYRAZOLO[1,5-a]PYRIMIDINE COMPOUNDS AS TRK KINASE INHIBITORS
WO2011146336A1 (en) 2010-05-20 2011-11-24 Array Biopharma Inc. Macrocyclic compounds as trk kinase inhibitors
WO2012116217A1 (en) 2011-02-25 2012-08-30 Irm Llc Compounds and compositions as trk inhibitors
WO2018077246A1 (zh) 2016-10-28 2018-05-03 正大天晴药业集团股份有限公司 用作神经营养因子酪氨酸激酶受体抑制剂的氨基吡唑并嘧啶化合物
WO2019118584A1 (en) * 2017-12-15 2019-06-20 Pyramid Biosciences, Inc. 5-(2-(2,5-difluorophenyl)pyrrolidin-1 -yl)-3-(1h-pyrazol-1-yl)pyrazolo[1,5-a]pyrimidine derivatives and related compounds as trk kinase inhibitors for treating cancer

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CUI J. ET AL., J. MED. CHEM., vol. 54, 2011, pages 6342 - 6363
DRILON A. ET AL., ANNALS OF ONCOLOGY, vol. 27, no. 5, 2016, pages 920 - 926
RUSSO M ET AL., CANCER DISCOVERY, vol. 6, no. 1, 2016, pages 36 - 44
See also references of EP3932923A4

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