WO2020156243A1 - Shp2 inhibitor and application thereof - Google Patents

Shp2 inhibitor and application thereof Download PDF

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Publication number
WO2020156243A1
WO2020156243A1 PCT/CN2020/072774 CN2020072774W WO2020156243A1 WO 2020156243 A1 WO2020156243 A1 WO 2020156243A1 CN 2020072774 W CN2020072774 W CN 2020072774W WO 2020156243 A1 WO2020156243 A1 WO 2020156243A1
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amino
compound
piperidine
dihydrospiro
indene
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PCT/CN2020/072774
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French (fr)
Chinese (zh)
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吴颢
吴文茂
陈忠研
李玲
林远望
吴云飞
朱林强
袁丁
王丰
张展
冯东杰
赵新涛
余军
束庆玉
程见洪
韩晗
郭晶
兰宏
王家炳
丁列明
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贝达药业股份有限公司
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Priority to CN202080008742.2A priority Critical patent/CN113365988B/en
Publication of WO2020156243A1 publication Critical patent/WO2020156243A1/en

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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/4965—Non-condensed pyrazines
    • A61K31/497—Non-condensed pyrazines containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
    • A61K31/5375—1,4-Oxazines, e.g. morpholine
    • A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/66—Phosphorus compounds
    • A61K31/675—Phosphorus compounds having nitrogen as a ring hetero atom, e.g. pyridoxal phosphate
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains two hetero rings
    • C07D471/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D471/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00
    • C07D471/12—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, at least one ring being a six-membered ring with one nitrogen atom, not provided for by groups C07D451/00 - C07D463/00 in which the condensed system contains three hetero rings
    • C07D471/20—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/107—Spiro-condensed systems with only one oxygen atom as ring hetero atom in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
    • C07D491/10—Spiro-condensed systems
    • C07D491/113—Spiro-condensed systems with two or more oxygen atoms as ring hetero atoms in the oxygen-containing ring
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
    • C07D491/12—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains three hetero rings
    • C07D491/20—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D495/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms
    • C07D495/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having sulfur atoms as the only ring hetero atoms in which the condensed system contains two hetero rings
    • C07D495/10—Spiro-condensed systems
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D513/00—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00
    • C07D513/02—Heterocyclic compounds containing in the condensed system at least one hetero ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for in groups C07D463/00, C07D477/00 or C07D499/00 - C07D507/00 in which the condensed system contains two hetero rings
    • C07D513/10—Spiro-condensed systems

Definitions

  • the present invention relates to a series of compounds as inhibitors of Src homology region 2 (Src homology region 2-containing protein tyrosine phosphatase 2, SHP2), and preparation methods and pharmaceutical compositions thereof.
  • the present invention also relates to the use of the above-mentioned compound or its pharmaceutical composition in the treatment of SHP2-mediated diseases.
  • Src homology region 2 protein tyrosine phosphatase 2 is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene, and PTPN11 is the first The discovered proto-oncogene encoding tyrosine kinase (Chan R J et al.PTPN11 is the first identified proto-oncogene that encodes a tyrosine phosphatase.Blood, 2007,109:862-867), and the encoded SHP2 protein contains N-terminal SHP2 domain (N-SHP2), C-terminal SHP2 domain (C-SHP2), protein phosphatase catalytic domain (PTP), two C-terminal tyrosine residues (Y542 and Y580) and one rich Proline (Pro) motif.
  • SOS as a guanine nucleotide exchange factor (GEF)
  • GEF guanine nucleotide exchange factor
  • Ras-GTP further connects with downstream signaling systems to activate Ser/Thr kinase Raf1, etc., and then activate ERK under the action of the regulatory kinase MEK. After activation of ERK, it directly acts on target molecules in the cytoplasm or transfers to the nucleus to regulate genes Transcription to make cells proliferate or differentiate. This process may also be affected by SHP2 binding protein and substrate (SHP substrate-1, SHPS-1), Ras-GTPase activating protein (Ras-GAP) and other members of Src.
  • SHP2 binding protein and substrate SHP substrate-1, SHPS-1
  • Ras-GTPase activating protein Ras-GTPase activating protein
  • SHP2 protein not only regulates the Ras/ERK signaling pathway, it is also reported to regulate multiple signaling pathways such as JAK-STAT3, NF- ⁇ B, PI3K/Akt, RHO, and NFAT, thereby regulating cell proliferation, differentiation, migration, and apoptosis Features.
  • SHP2 has been proved to be related to many diseases, Tartaglia et al. (Tartaglia M et al. Mutations in PTPN11, encoding the protein tyrosine phosphatase SHP-2, cause Noonan syndrome. Nat Genet, 2001, 29:465-468) found that about 50% Patients with Noonan syndrome have missense mutations of PTPN11. In addition, studies have found that PTPN11 mutation is an important cause of JMML and a variety of leukemias (Tartaglia M et al. Nat Genet, 2003, 34: 148-150; Loh ML et al. Blood, 2004, 103: 2325-2331; Tartaglia M et al.
  • SHP2 inhibitors have received more and more attention as potential treatments.
  • the present invention relates to a compound used as an inhibitor of protein tyrosine phosphatase 2 (SHP2) containing Src homology region 2.
  • SHP2 protein tyrosine phosphatase 2
  • the compound of the present invention has a general structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,
  • R 1 is selected from amino, -C (O) -R a, -C ⁇ N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, C 1-8 alkoxy containing substituents;
  • R a is optionally selected from amino, -NH-OH, C 1-3 alkyl
  • R 2 is selected from hydrogen, C 1-4 alkyl group or a substituted C 1-4 alkyl
  • R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C ⁇ N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
  • R 4 is selected from hydrogen, halo, amino, amido, -C ⁇ N, carboxyl, hydroxyl, hydroxymethyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl base;
  • a 1 is arbitrarily selected from CR 5 or N;
  • a 2 is arbitrarily selected from CR 6 or N;
  • a 3 is arbitrarily selected from CR 7 or N;
  • a 4 is arbitrarily selected from CR 8 or N;
  • U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
  • R 5 , R 6 , R 7 , R 8 , R 9 or R 10 are independently selected from hydrogen, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted C 1 -8 alkyl, C 1-8 alkoxy, substituted C 1-8 alkoxy, C 2-8 alkenyl, substituted C 2-8 alkenyl, C 2-8 alkynyl, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents; or,
  • L is selected from S
  • Ring A is optionally selected from a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group contains one or two N or S heteroatoms;
  • Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;
  • n 0, 1, 2, 3, or 4.
  • R 1 in Formula I optionally selected from amino, -C (O) NH 2, -C ⁇ N, C 1-3 alkyl group, a substituted C 1-3 alkyl group or a C 1 -3 Alkoxy.
  • R 1 in Formula I is arbitrarily selected from amino, -C(O)NH 2 , -C ⁇ N, methyl substituted by hydroxy,
  • R 2 in Formula I is hydrogen
  • R 3 of Formula I is optionally selected from hydrogen, halo, C 1-3 alkyl or an optionally substituted C 1-3 alkyl.
  • R 3 in Formula I is arbitrarily selected from hydrogen, chlorine, or methyl.
  • R 4 in Formula I is arbitrarily selected from hydrogen, halogen, C 1-8 alkyl or C 1-8 alkoxy containing substituents.
  • R 4 in Formula I is arbitrarily selected from hydrogen, F, Cl, -CHF 2 , CF 3 or -O-CH 3 .
  • a 1 in formula I is CR 5 or N, wherein R 5 is selected from hydrogen, halogen, or halogen substituted C 1-3 alkyl.
  • a 1 in formula I is CR 5 or N, wherein R 5 is selected from hydrogen, Cl or trifluoromethyl.
  • a 2 in Formula I is CR 6 or N, wherein R 6 is selected from hydrogen, hydroxyl, halogen, amino, or C 1-8 alkoxy.
  • a 2 in Formula I is CR 6 or N, wherein R 6 is selected from hydrogen, OH, F, Cl, amino, or -O-CH 3 .
  • a 3 in formula I is CR 7 or N, wherein R 7 is hydrogen or halogen.
  • a 4 in formula I is CR 8 or N, wherein R 8 is selected from hydrogen, halogen, amino, substituted amino, C 1-3 alkoxy, or C 5-6 heterocyclyl.
  • a 4 in formula I is CR 8 or N, wherein R 8 is selected from hydrogen, F, Cl, amino, -NHCH 3 , -N(CH 3 ) 2 , Or methoxy.
  • U in Formula I is CH 2 or O.
  • ring A in formula I is arbitrarily selected from phenyl or C 5-6 heteroaryl, and the C 5-6 heteroaryl contains one or two N or S heteroatoms.
  • Rx in Formula I is arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C 1-3 alkyl, halogen-substituted C 1-3 alkyl, or C 1-3 alkoxy.
  • Rx in Formula I is arbitrarily selected from hydrogen, OH, F, Cl, Br, -CN, trifluoromethyl, or methoxy.
  • the present invention further provides some preferred technical solutions of the compound represented by formula I.
  • the compound of the present invention has the general structure shown in Formula II or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,
  • R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C ⁇ N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
  • a 1 is arbitrarily selected from CR 5 or N;
  • U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
  • R 5 , R 9 or R 10 are independently selected from hydrogen, hydroxy, halogen, amino, substituted amino, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl Group or C 5-6 heterocycle; or,
  • L is selected from S
  • R x is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy group, a substituted group containing C 1-8 alkoxy, C 3-8 cycloalkyl or C 3-8 cycloalkyl group having substituent;
  • n 0, 1, 2, 3, or 4.
  • R 3 in Formula II is optionally selected from hydrogen or C 1-3 alkyl.
  • R 3 in Formula II is arbitrarily selected from hydrogen or methyl.
  • a 1 in formula II is CR 5 or N, wherein R 5 is selected from halogen.
  • a 1 in Formula II is CR 5 , wherein R 5 is selected from F or Cl.
  • a 1 in Formula II is N.
  • U in Formula II is O or CH 2 .
  • R x in Formula II is arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C 1-3 alkyl, halogen-substituted C 1-3 alkyl, or C 1-3 alkoxy.
  • R x in Formula II is arbitrarily selected from hydrogen, OH, CN, Cl, F, Br, trifluoromethyl, or methoxy.
  • n in Formula II is 0, 1, or 2.
  • the present invention further provides some preferred technical solutions of the compound represented by formula I.
  • the compound of the present invention has the general structure shown in Formula III or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,
  • R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C ⁇ N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
  • R 4 is selected from hydrogen, halo, amino, amido, -C ⁇ N, carboxyl, hydroxyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, A substituted C 1-8 alkoxy group, a C 2-8 alkenyl group, a substituted C 2-8 alkenyl group, a C 2-8 alkynyl group or a substituted C 2-8 alkynyl group;
  • a 1 is arbitrarily selected from CR 5 or N;
  • a 2 is arbitrarily selected from CR 6 or N;
  • a 3 is arbitrarily selected from CR 7 or N;
  • U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
  • R 5 , R 6 , R 7 , R 8 , R 9 or R 10 are independently selected from hydrogen, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted C 1 -8 alkyl, C 1-8 alkoxy, substituted C 1-8 alkoxy, C 2-8 alkenyl, substituted C 2-8 alkenyl, C 2-8 alkynyl, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents;
  • L is selected from S
  • Ring A is optionally selected from a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group contains one or two N or S heteroatoms;
  • Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;
  • n 0, 1, 2, 3, or 4.
  • R 3 in Formula III is selected from hydrogen.
  • R 4 in Formula III is arbitrarily selected from hydrogen or Cl.
  • a 1 in Formula III is CR 5 or N, wherein R 5 is selected from hydrogen, Cl or trifluoromethyl.
  • a 2 in formula III is CR 6 or N, wherein R 6 is selected from Cl, amino, Or -O-CH 3 .
  • a 3 in Formula III is CR 7 or N, wherein R 7 is hydrogen.
  • U in Formula III is CH 2 .
  • ring A in formula III is arbitrarily selected from phenyl or C 5-6 heteroaryl, and the C 5-6 heteroaryl contains one or two N or S heteroatoms.
  • Rx in Formula III is arbitrarily selected from hydrogen, Cl, Br, trifluoromethyl, or methoxy.
  • the present invention further provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:
  • the present invention also provides a pharmaceutical composition characterized by comprising a therapeutically effective amount of at least one compound represented by formula (I) and at least one pharmaceutically acceptable excipient.
  • the present invention further provides a pharmaceutical composition, characterized in that the mass percentage of the therapeutically effective amount of at least one compound represented by formula (I) and pharmaceutically acceptable excipients is 0.0001:1-10.
  • the invention provides the application of the compound or pharmaceutical composition represented by the structural formula (I) in the preparation of medicines.
  • the application is an application for preparing a medicine for treating, preventing, delaying or preventing cancer, cancer metastasis, cardiovascular disease, immune disease, fibrosis or eye disease.
  • the application is an application in preparing a medicine for treating diseases mediated by SHP2.
  • the disease is cancer.
  • the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal tumor , Lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma, pancreatic cancer or a combination thereof.
  • the application is an application for preparing an SHP2 inhibitor.
  • the present invention also provides a method for treating and/or preventing diseases mediated by SHP2, which comprises administering a therapeutically effective amount of at least any one compound or pharmaceutical composition represented by structural formula (I) to a subject.
  • the SHP2-mediated disease is cancer.
  • the present invention also provides a method for treating cancer, which comprises administering a therapeutically effective amount of at least any one compound or pharmaceutical composition represented by structural formula (I) to a subject.
  • the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal tumor , Lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma, pancreatic cancer or a combination thereof.
  • the treatment target is a human.
  • alkyl includes straight, branched or cyclic saturated alkyl groups.
  • alkyl includes but is not limited to methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, 3 -(2-Methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, cyclopentyl, n-hexyl, 2-hexyl, 2-methylpentyl and cyclohexyl and other similar groups group.
  • C 1-8 alkyl means comprising 7 or 8 carbon atoms, a straight chain, branched chain or cyclic Formally arranged groups.
  • alkenyl and alkynyl include linear, branched or cyclic alkenyl and alkynyl groups.
  • C 2-8 alkenyl and “C 2-8 alkynyl” refer to a linear, branched or cyclic arrangement containing 2, 3, 4, 5, 6, 7 or 8 carbon atoms ⁇ alkenyl or alkynyl.
  • alkoxy refers to the oxyether form of the aforementioned linear, branched or cyclic alkyl group.
  • aryl refers to an unsubstituted or substituted monocyclic or polycyclic aromatic group including carbon atoms. It is preferably a 6 to 10 membered monocyclic or bicyclic aromatic group. Preferably it is phenyl and naphthyl. Most preferred is phenyl.
  • heteroaryl refers to a monovalent heteroatom group formed by removing a hydrogen atom from a carbon atom of a parent heteroaromatic ring system.
  • Heteroaryl groups include: 5- to 7-membered aromatic, monocyclic, including at least one heteroatom selected from N, O or S, for example, 1 to 4 heteroatoms, or preferably 1 to 3 heteroatoms, ring The other atoms on the above are carbon; the polyheteroaryl ring includes at least one heteroatom selected from N, O or S, for example, 1 to 4 heteroatoms, or preferably 1 to 3 heteroatoms, other atoms in the ring Is carbon, and at least one of the heteroatoms is on the aromatic ring.
  • heteroaryl groups are C 3-10 heteroaryl groups, including but not limited to, pyrrolyl, furyl, thienyl, pyridyl, pyranyl, pyrazolyl, pyrimidinyl, pyridazinyl, Pyrazinyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, triazole, indolyl, benzofuranyl, benzothiazolyl, benzimidazolyl, benzopyrazolyl, benzene And similar groups such as triazolyl, carbazolyl, quinolinyl, isoquinolinyl, purinyl and the like.
  • the heteroaryl group and the aryl group will not cross or contain each other. Therefore, according to the above definition, if at least one all-carbon aromatic ring is fused with a heterocyclic group, the result is a heteroaryl group instead of an aryl group.
  • Cycloalkyl refers to a cyclic group that is saturated or unsaturated but not aromatic. Depending on the specific level of saturation, the terms “cycloalkyl”, “cycloalkenyl” or “cycloalkynyl” are used respectively.
  • Representative cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclohexane, or cyclohexene and similar groups.
  • the cycloalkyl group may be a C 3-10 cycloalkyl group, such as a C 3-6 cycloalkyl group.
  • Heterocyclic group refers to a saturated or unsaturated but not aromatic cyclic group, and one or more of the carbon atoms (and the connected hydrogen atoms) can be respectively the same or different hetero Atom and the corresponding attached hydrogen atom are replaced.
  • Representative heteroatoms that replace carbon atoms include, but are not limited to, N, P, O, S, and Si. When a specific degree of saturation needs to be described, the terms “heterocycloalkyl” or “heterocycloalkenyl” are used respectively.
  • heterocyclic groups include but are not limited to epoxy compounds, imidazolidine, morpholine, piperazine, piperidine, pyrazolidine, pyrrolidine, quinuclidine, tetrahydrofuran or tetrahydropyran and similar groups .
  • the heterocycloalkyl group and the cycloalkyl group will not cross or contain each other. Therefore, according to the above definition, if at least one all-carbocyclic ring is fused with a heterocycloalkyl to form a di-, poly- or spiro-ring, it will still be defined as a heterocycloalkyl.
  • heteroaryl group is fused with a heterocyclic group to form a di-, poly- or spiro-ring, it will be defined as a heterocyclic group instead of a heteroaryl group.
  • Halogen refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). Preferred halogen refers to fluorine, chlorine and bromine.
  • Halo refers to a fluoro, chloro, bromo or iodo group.
  • the preferred halo groups refer to fluoro and chloro.
  • substitution means that one or more hydrogen atoms in a group are replaced by the same or different substituents.
  • substituents include, but are not limited to, halogen, amino, hydroxy, oxo, carbonyl, cyano, -C (O) NH 2, alkyl, alkoxy, aryl, cycloalkyl, heterocyclyl , Heteroaryl.
  • the substituent includes, but is not limited to, halogen, amino, hydroxy, cyano, methyl, -CH 2 OH, -C(O)NH 2 , -OCH 3 , trifluoromethyl.
  • alkyl or aryl or its prefixes appear in the name of a substituent (such as aralkyl, or dialkylamino), it shall be the same as the aforementioned "alkyl” and "aryl”"The definition defines the substituents.
  • the specified number of carbon atoms (such as C 1-6 ) will independently represent the number of carbon atoms in an alkyl moiety or in an alkyl moiety (where the alkyl group is the prefix stem) in a larger substituent.
  • the "compound” of the present invention includes the compound represented by formula (I), and all pharmaceutically acceptable forms thereof. These pharmaceutically acceptable forms include salts, solvates, non-covalent complexes, chelates or their prodrugs, or any mixture of all the above forms.
  • the “pharmaceutically acceptable” refers to those that are well known for use in animals, especially those that can be used in humans.
  • composition in the present invention includes a product containing a specific quantity of a specific component, and also includes any product directly or indirectly obtained from a specific quantity of a specific component. Therefore, a pharmaceutical composition including the compound of the present invention as an active ingredient and a method for preparing the compound are the content of the present invention.
  • “Therapeutically effective amount” means that when a compound is administered to a subject to treat and prevent and/or inhibit at least one clinical symptom of a disease, condition, symptom, indication, and/or discomfort, it is sufficient for the disease, condition, A dose that produces a certain effect in the treatment of symptoms, indications or discomfort.
  • the specific "effective therapeutic dose” may vary according to the compound, the route of administration, the age of the patient, the weight of the patient, the type of disease or discomfort to be treated, the symptoms and severity, etc. Whenever possible, an appropriate dose may be obvious to those skilled in the art, or it may be determined by conventional experimental methods.
  • the compounds provided by the present invention may exist in the form of "pharmaceutically acceptable salts".
  • the salt of the compound provided by the present invention refers to a non-toxic pharmaceutically acceptable salt.
  • the form of the pharmaceutically acceptable salt includes a pharmaceutically acceptable acid/anion or base/cation salt.
  • Pharmaceutically acceptable acid/anionic salts generally exist in the form of protonation of basic nitrogen with inorganic or organic acids.
  • Typical organic or inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, perchloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, lactic acid, succinic acid, maleic acid, fumaric acid, malic acid , Tartaric acid, citric acid, ⁇ -ketoglutaric acid, hippuric acid, benzoic acid, mandelic acid, methanesulfonic acid, isethionic acid, benzenesulfonic acid, oxalic acid, pamoic acid, 2-naphthalenesulfonic acid, p-toluenesulfonic acid Acid, cyclohexylamine sulfonic acid, salicylic acid, saccharic acid or trifluoroacetic acid.
  • Pharmaceutically acceptable base/cation salts including, but not limited to, aluminum salt, calcium salt, chloroprocaine salt, choline, diethanolamine salt
  • the prodrug of the compound of the present invention is included in the protection scope of the present invention.
  • the prodrug is a functional derivative that is easily converted into a desired compound in vivo. Therefore, the term "administration" involved in the treatment method provided by the present invention includes the administration of the compound disclosed in the present invention, or although it is not clearly disclosed but can be transformed into the compound disclosed in the present invention in vivo after administration to the subject. disease.
  • the conventional methods for selecting and preparing suitable prodrug derivatives have been recorded in books such as "Design of Prodrugs” (Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985).
  • any substituent or variable at a specific position in one molecule is irrelevant to the definition of any substituent or variable at a specific position in other molecules. It is easy to understand that the compounds of the present invention can be selected according to the prior art of the subject to select suitable substituents or substitution forms to provide chemically stable and easy preparation and synthesis using the prior art of the subject or the method described in the present invention.
  • the present invention includes any possible solvate and polymorph.
  • the type of solvent that forms the solvate is not particularly limited, as long as the solvent is pharmacologically acceptable.
  • water, ethanol, propanol, acetone and similar solvents can be used.
  • pharmaceutically acceptable salt refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid.
  • pharmaceutically acceptable non-toxic bases including inorganic bases and organic bases.
  • Salts derived from inorganic bases include aluminum, ammonium, calcium, copper (ic and ous), iron, ferrous, lithium, magnesium, manganese (ic and ous), potassium, sodium, zinc and the like. In particular, salts of ammonium, calcium, magnesium, potassium, and sodium are preferred.
  • Non-toxic organic bases that can be derivatized into pharmaceutically acceptable salts include primary, secondary and tertiary amines, as well as cyclic amines and amines containing substituents, such as naturally occurring and synthetic amines containing substituents.
  • non-toxic organic bases capable of forming salts, including ion exchange resins and arginine, betaine, caffeine, choline, N',N'-dibenzylethylene diamine, diethylamine, 2 -Diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, reduced glucosamine, glucosamine, histidine, haamine, isopropylamine , Lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.
  • the corresponding salt can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids and organic acids.
  • acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, isethionic acid, formic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, Hydrochloric acid, lactic acid, maleic acid, malic acid, mandelic acid, ⁇ -ketoglutaric acid, hippuric acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonate Acid etc.
  • malic acid citric acid, hydrobromic acid, hydrochloric acid, methanesulfonic acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid. More preferably, phosphoric acid, hydrochloric acid and malic acid. Since the compound represented by formula (I) will be used as a medicine, it is preferable to use a substantially pure form, for example, at least 60% purity, more suitably at least 75% purity, particularly suitably at least 98% purity (% is weight ratio) .
  • the pharmaceutical composition provided by the present invention includes a compound represented by formula (I) (or a pharmaceutically acceptable salt thereof) as an active component, a pharmaceutically acceptable excipient and other optional therapeutic components or Accessories.
  • the pharmaceutical composition of the present invention includes oral, rectal, topical and Pharmaceutical composition for parenteral (including subcutaneous administration, intramuscular injection, intravenous administration) administration.
  • the pharmaceutical composition of the present invention can be conveniently prepared in a unit dosage form known in the art and prepared by any preparation method known in the pharmaceutical field.
  • the compound represented by formula (I) of the present invention can be used in combination as an active ingredient and mixed with a drug carrier to form Pharmaceutical composition.
  • the pharmaceutical carrier can take various forms, depending on the desired mode of administration, for example, oral or injection (including intravenous injection). Therefore, the pharmaceutical composition of the present invention may take the form of a separate unit suitable for oral administration, such as a capsule, cachet or tablet containing a predetermined dose of the active ingredient.
  • the pharmaceutical composition of the present invention may take the form of powder, granule, solution, aqueous suspension, non-aqueous liquid, oil-in-water emulsion, or water-in-oil emulsion.
  • the compound represented by formula (I) or a pharmaceutically acceptable salt thereof can also be administered through a controlled release method and/or a delivery device.
  • the pharmaceutical composition of the present invention can be prepared by any pharmaceutical method. Generally, this method includes the step of associating the active ingredient with the carrier constituting one or more necessary ingredients.
  • the pharmaceutical composition is prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a finely divided solid carrier or a mixture of both.
  • the product can be easily prepared into the desired appearance.
  • the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier and a compound represented by formula (I), or a pharmaceutically acceptable salt thereof.
  • the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, and one or more other compounds with therapeutic activity in combination are also included in the pharmaceutical composition of the present invention.
  • the drug carrier used in the present invention can be, for example, a solid carrier, a liquid carrier or a gas carrier.
  • solid carriers include lactose, gypsum powder, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, mannitol, sorbitol, microcrystalline cellulose, inorganic salts , Starch, pregelatinized starch, powdered sugar, dextrin, etc.
  • liquid carriers include syrup, peanut oil, olive oil and water.
  • gas carriers include carbon dioxide and nitrogen.
  • water, ethylene glycol, oils, alcohols, flavor enhancers, preservatives, coloring agents, etc. can be used for oral liquid preparations such as suspensions, elixirs and solutions; and carriers, such as starches, sugars, Microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc. can be used in oral solid preparations such as powders, capsules and tablets. In view of ease of administration, tablets and capsules are preferred for oral preparations. Alternatively, standard aqueous or non-aqueous formulation techniques can be used for tablet coating.
  • Tablets containing the compound or pharmaceutical composition of the present invention can be prepared by, optionally, mixing, compression or molding with one or more auxiliary components or adjuvants.
  • the active ingredient is in a free-flowing form such as powder or granules, mixed with lubricants, inert diluents, surface active or dispersing agents, and compressed in a suitable machine to obtain compressed tablets.
  • the powdered compound or pharmaceutical composition is wetted with an inert liquid diluent, and then molded in a suitable machine to form a molded tablet.
  • each tablet contains about 0.01 mg to 5 g of active ingredient, and each cachet or capsule contains about 0.1 mg to 0.5 g of active ingredient.
  • a dosage form intended for oral administration to humans contains about 0.1 mg to about 0.5 g of the active ingredient, compounded with a suitable and convenient metering auxiliary material, which accounts for about 5% to 99.99% of the total pharmaceutical composition .
  • the unit dosage form generally contains about 0.1mg to about 0.5g of effective ingredients, typically 0.1mg, 0.2mg, 0.5mg, 1mg, 2mg, 2.5mg, 5mg, 10mg, 25mg, 50mg, 100mg, 200mg, 300mg, 400mg Or 500mg.
  • the pharmaceutical composition suitable for parenteral administration provided by the present invention can be prepared as an aqueous solution or suspension by adding active components into water.
  • Suitable surfactants such as sodium lauryl sulfate, polysorbate-80 (Tween-80), polyoxyethylene hydrogenated castor oil, and poloxamer may be included.
  • glycerol liquid polyethylene glycol, and their mixtures in oil, dispersion systems can also be prepared.
  • a preservative may also be included in the pharmaceutical composition of the present invention to prevent the growth of harmful microorganisms.
  • the present invention provides pharmaceutical compositions suitable for injection use, including sterile aqueous solutions or dispersion systems.
  • the above-mentioned pharmaceutical composition can be prepared in the form of a sterile powder that can be used for immediate preparation of sterile injection.
  • the final injection form must be sterile, and for easy injection, it must be easy to flow.
  • the pharmaceutical composition must be stable during preparation and storage. Therefore, preservation against contamination by microorganisms such as bacteria and fungi is preferred.
  • the carrier can be a solvent or dispersion medium, for example, water, ethanol, polyol (such as glycerol, propylene glycol, liquid polyethylene glycol), vegetable oil, and suitable mixtures thereof.
  • the pharmaceutical composition provided by the present invention may be in a form suitable for topical administration, for example, aerosol, emulsion, ointment, lotion, dusting, or other similar dosage forms. Further, the pharmaceutical composition provided by the present invention can be in a form suitable for use in a transdermal drug delivery device.
  • These preparations can be prepared by using the compound represented by the formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, through conventional processing methods.
  • an emulsion or ointment is prepared by adding a hydrophilic material and water (the total amount of the two is about 5 wt% to 50 wt% of the compound) to prepare a cream or ointment with the desired consistency.
  • the pharmaceutical composition provided by the present invention can be made into a form suitable for rectal administration with a solid as a carrier.
  • Suppositories in which the mixture forms a unit dose are the most preferred dosage form.
  • Suitable excipients include cocoa butter and other materials commonly used in the art. Suppositories can be conveniently prepared. First, the pharmaceutical composition is mixed with softened or melted excipients, then cooled and molded.
  • the above-mentioned pharmaceutical preparations may also include, as appropriate, one or more additional adjuvant components, such as diluents, buffers, flavoring agents, binders, surfactants, and additives. Thickeners, lubricants, preservatives (including antioxidants), etc. Further, other adjuvants may also include penetration enhancers that regulate the isotonic pressure of the drug and blood.
  • additional adjuvant components such as diluents, buffers, flavoring agents, binders, surfactants, and additives. Thickeners, lubricants, preservatives (including antioxidants), etc.
  • other adjuvants may also include penetration enhancers that regulate the isotonic pressure of the drug and blood.
  • the pharmaceutical composition containing the compound represented by formula (I), or a pharmaceutically acceptable salt thereof can also be prepared in the form of a powder or a concentrated solution.
  • ACE-Cl 1-chloroethyl chloroformate
  • BOP Benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate
  • DBU 1,8-diazabicycloundec-7-ene
  • DIPEA or DIEA N,N-diisopropylethylamine
  • DMSO dimethyl sulfoxide
  • EDCI 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride
  • LDA lithium diisopropylamide
  • MsCl methanesulfonyl chloride
  • NEt 3 Triethylamine
  • NBS N-bromosuccinimide
  • NMP N-methyl-2-pyrrolidone
  • Pd 2 (dba) 3 Tris(dibenzylideneacetone)dipalladium
  • PE petroleum ether
  • PPA polyphosphoric acid
  • rt, r.t. or RT room temperature
  • TEA triethylamine
  • TFA trifluoroacetic acid
  • THF Tetrahydrofuran
  • Ti(OEt) 4 Tetraethyl titanate
  • TMEDA Tetramethylethylenediamine
  • xantphos 4,5-bisdiphenylphosphine-9,9-dimethylxanthene.
  • the compound M5-3 was dissolved in 100 mL DMF, the temperature was lowered to 0°C, 4.98 g NBS was added all at once, and the reaction was carried out at RT for 1 hr.
  • the reaction was completed by TLC detection, the reaction was quenched by adding 300 mL of water, extracted with EtOAc (300 mL ⁇ 2), combined the organic phases, washed with saturated NaCl (50 mL ⁇ 4), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography 5.58 g of white solid, compound M5 was obtained.
  • the procedure for preparing compound M7 from compound M7-7 is similar to the procedure from compound M6-3 to compound M6.
  • the procedure for preparing M9-a and M9-b from M9-8a and M9-8b is similar to the procedure from compound M6-3 to compound M6.
  • the procedure for preparing compound M10 from compound M10-4 is similar to the procedure from compound M6-3 to compound M6.
  • the starting material for the synthesis of M12 is 6-methoxy-1-indanone
  • the intermediate compounds in Table 1 are prepared by a method similar to the aforementioned intermediate 6-10 M11-M15.
  • Example 2 Compound (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3- Preparation of dichlorophenyl)thio)-5-methylpyrazin-2-yl)methanol
  • Example 3 Compound (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazine-2-carbonitrile preparation
  • Example 4 Compound (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-bis Preparation of chlorophenyl)thio)pyrazine-2-carboxamide
  • Example 5 Compound (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazine-2-carboxamide
  • the compounds 7-18, 58 and 59 in Table 2 were prepared by using different reaction starting materials and appropriate reagents using a method similar to that of the foregoing Example 5.
  • an aqueous solution (25 mL) of sodium hydroxide (546 mg) was added dropwise to an aqueous solution in which 1.00 g of compound M25-1, 2.60 g of elemental iodine and 1.70 g of potassium iodide were dissolved.
  • the reaction was carried out at room temperature for 2 hours, and the reaction was completed as detected by TLC. Adjust the pH to neutral with saturated ammonium chloride solution. Then use sodium thiosulfate solution to remove elemental iodine. It was extracted with methyl tert-butyl ether (50 mL ⁇ 2), and the organic layers were combined, dried with sodium sulfate and desolvated. 1.45 g of compound M25-2 was obtained.
  • reaction solution was quenched with 200 mL saturated brine, extracted with EtOAc (300 mL ⁇ 2), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 11.06 g of compound M27-5.
  • TLC detects that the reaction is complete, concentrate, add 50mL ethyl acetate and 50mL water to disperse, add 50mL ethyl acetate to extract the aqueous phase, combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, the residue is purified by column chromatography to obtain a class 0.605 g of compound 22-2 as a white solid.
  • the compounds 28-57 and 63 in Table 3 were prepared by using different reaction starting materials and appropriate reagents, using a method similar to that of the foregoing Example 26.
  • SHP2 is allosterically activated by the binding of a bis-tyrosyl-phosphorylated peptide to its Src homology 2 (SH2) domain.
  • SH2 Src homology 2
  • the subsequent activation step results in the release of the SHP2 auto-inhibitory interface, which in turn activates the SHP2 protein tyrosine phosphatase (PTP) and can be used for substrate recognition and reaction catalysis.
  • PTP protein tyrosine phosphatase
  • the surrogate DiFMUP was used to monitor the catalytic activity of SHP2 in the rapid fluorescence assay format.
  • the compound of the present invention (10mM stock solution) was diluted to an appropriate multiple with 100% DMSO.
  • the final test concentration of the compound of the present invention was 10 ⁇ M, 3.3333 ⁇ M, 1.1111 ⁇ M, 0.3704 ⁇ M, 0.1235 ⁇ M, 0.0412 ⁇ M, 0.0137 ⁇ M, 0.0046 ⁇ M, 0.0015 ⁇ M , 0.00 ⁇ M;
  • Inhibition rate% [1-(Conversion_ sample -Conversion_ min )/(Conversion_ max -Conversion_ min )] ⁇ 100%
  • Conversion_sample is the conversion rate reading of the sample
  • Conversion_min is the average value of the blank control well, representing the conversion rate reading of the wells without enzyme activity
  • Conversion_max is the average value of the positive control well ratio, representing the conversion rate reading of the wells without compound inhibition.
  • the analysis software GraphPad Prism log (inhibitor) vs. response-Variable slope was used to fit the dose-response curve, and the IC 50 value of the compound to the enzyme activity was calculated.
  • the IC 50 data of some examples are shown in Table 4.
  • Compound 32 1.47 Compound 33 0.9 Compound 34 1.7 Compound 35 0.7 Compound 36 1.19 Compound 37 1.2 Compound 38 0.6 Compound 39 0.71 Compound 40 1.80 Compound 41 0.81 Compound 42 0.85 Compound 43 2.41 Compound 44 0.83 Compound 45 1.60 Compound 46 1.10 Compound 47 0.60 Compound 48 1.7 Compound 49 4.1 Compound 50 0.7 Compound 51 / Compound 52 / Compound 53 2 Compound 54 1 Compound 55 9.3 Compound 56 6.8 Compound 57 / Compound 58 0.72 Compound 59 2.15 Compound 60 7.19 Compound 61 19.75 Compound 62 1.4 Compound 63 4.3 Compound 64 2.0 Compound 65 2.1 Compound 66 3.1 Compound 67 2.0 Compound 68 1.8 Compound 69 4.7 Compound 70 2.3
  • the compound of the present invention has an allosteric inhibitory effect on SHP2 protein.
  • An in vitro cell assay was used to evaluate the effects of the compounds of the present invention on the proliferation of lung squamous cell carcinoma KYSE-520 cells and leukemia cells MV-4-11 cells.
  • the detection method used in the experiment is the CELL TITER-GLO (CTG) luminescence method, which can detect the number of living cells by quantitatively measuring ATP. Because ATP participates in a variety of enzymatic reactions in organisms, it is an indicator of living cell metabolism. Its content directly reflects the number and cell state of cells.
  • CCG CELL TITER-GLO
  • Inhibition rate% (1-(administration group value-zero adjustment group value)/(blank group value-zero adjustment group value)*100
  • the compound of the present invention has a good inhibitory effect on the proliferation of KYSE-520 cells and the proliferation of MV-4-11 cells.
  • Example C Inhibition test of hERG potassium ion channel
  • the whole cell patch clamp technique was used to detect the blocking effect of the test compound on the hERG channel.
  • the HEK293 cell line stably expressing the hERG potassium channel was cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg/mL G418 at a culture temperature of 37°C and a carbon dioxide concentration of 5%.
  • the cell density In order to maintain the electrophysiological activity of cells, the cell density must not exceed 80%.
  • the cells were separated with TrypLE TM Express before the experiment, and 3*103 cells were spread on a cover glass and cultured in a 24-well plate (final volume: 500 ⁇ L). After 18 hours, the experiment was performed.
  • Cisapride positive control
  • the weighed 1.2mg cisap must be prepared into a 10mM stock solution using 243 ⁇ L DMSO.
  • the cisapride stock solution was diluted successively with DMSO at a 10-fold dilution from high to low to a 10 ⁇ M dilution.
  • the whole cell patch clamp voltage stimulation scheme for recording the whole cell hERG potassium current is as follows: when the whole cell seal is formed, the cell membrane voltage is clamped to -80mV. The clamping voltage is depolarized from -80mV to -50mV for 0.5 seconds, then stepped to 30mV for 2.5 seconds, and then quickly restored to -50mV for 4 seconds to stimulate the tail current of the hERG channel. Collect data repeatedly every 10 seconds to observe the effect of the drug on the hERG tail current. A stimulus of -50mV for 0.5 seconds was used as leakage current detection. The experimental data is collected by EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.
  • HEKA EPC-10 amplifier
  • HEKA PatchMaster
  • the capillary glass tube is drawn into a recording electrode with a microelectrode drawing instrument. Operate the microelectrode manipulator under the inverted microscope to contact the recording electrode on the cell, and apply negative pressure suction to form a G ⁇ seal. After forming the G ⁇ seal, perform fast capacitance compensation, and then continue to give negative pressure to suck and break the cell membrane to form a whole-cell recording mode. Then perform slow capacitance compensation and record the film capacitance and series resistance. No leakage compensation is given.
  • the drug When the hERG current recorded by the whole cell stabilizes, the drug is administered, and each drug concentration acts for 5 minutes (or the current stabilizes). Place the cover glass covered with cells in the recording bath in the inverted microscope.
  • the test compound and the compound-free external fluid flow through the recording bath sequentially by gravity perfusion to act on the cells, and the vacuum pump is used to carry out the liquid during recording. exchange.
  • the current detected by each cell in the compound-free external fluid serves as its own control group. Test multiple cells independently and repeatedly. All electrophysiological experiments were performed at room temperature.
  • the compounds of the present invention generally have good inhibitory activity on SHP2, when one of the following two conditions is met, namely 1), the position of R 1 on the pyrazine ring in formula I is a hydroxyl group. A methyl group and both A 1 and A 3 positions are N, such as compound 22, compounds 64-69; 2), in formula I, the R 1 position on the pyrazine ring is an amide group and A 3 position is N, such as compound 5, Compared with other technical solutions, compound 18, compound 58 or compound 59 can significantly improve the hERG inhibition problem and is expected to reduce cardiotoxicity.
  • Example D Plasma protein binding assay
  • An alkaline solution was prepared by dissolving 14.2 g/L Na 2 HPO 4 and 8.77 g/L NaCl in deionized water, which can be stored at 4° C. for up to 7 days.
  • An acidic solution can be prepared by dissolving 12.0g/L NaH 2 PO 4 and 8.77g/L NaCl in an acidic solution, which can be stored at 4°C for 7 days.
  • the alkaline solution was titrated to pH 7.4 with an acidic solution and stored at 4°C for 7 days. Check on the day of the experiment and adjust if the pH exceeds the specification of 7.4 ⁇ 0.1.
  • the frozen plasma was immediately thawed at room temperature.
  • the plasma was centrifuged at 3,220g for 10 minutes to remove clots, and the supernatant was collected in a new test tube. Check and record the pH of the plasma.
  • Working solutions of the test compound and the control compound ketoconazole were prepared with DMSO at a concentration of 400 ⁇ M. Then 4 ⁇ L of the working solution was removed and mixed with 796 ⁇ L of human, dog, monkey, rat, or mouse plasma to finally obtain a mixed solution with a concentration of 2 ⁇ M (0.5% DMSO). Vortex the plasma sample thoroughly.
  • % Free (peak area ratio buffer chamber /peak area ratio plasma chamber )*100
  • Peak area ratio buffer chamber indicates the concentration of free part
  • Peak area ratio plasma chamber indicates the concentration of free and bound fraction
  • the peak area ratio to the total sample indicates the concentration of the sample started before incubation
  • Table 7 shows the plasma protein binding results of the control compound and the test compound in different species.
  • the comparative example reflects a high degree of binding to plasma proteins, so the efficacy of the drug may be reduced.
  • the exemplary compound of the present invention has a lower degree of plasma protein binding compared with the comparative example. It is predicted that the present invention has excellent pharmacokinetics and pharmacodynamic properties on the human body.

Abstract

Provided is a compound for use as an inhibitor of Src homology region 2-containing protein tyrosine phosphatase 2 (SHP2) (as represented by formula I), and pharmaceutical composition and preparation method thereof, and use thereof in the treatment of SHP2-mediated diseases. The compound works by means of participating in various processes, such as regulating cell proliferation, apoptosis, migration, and neovascularization.

Description

SHP2抑制剂及其应用SHP2 inhibitor and its application 技术领域Technical field
本发明涉及一系列作为含Src同源区2蛋白质酪氨酸磷酸酶2(Src homologyregion 2-containing protein tyrosine phosphatase 2,SHP2)抑制剂的化合物及其制备方法、药物组合物。本发明还涉及上述化合物或其药物组合物在治疗SHP2介导的疾病中的用途。The present invention relates to a series of compounds as inhibitors of Src homology region 2 (Src homology region 2-containing protein tyrosine phosphatase 2, SHP2), and preparation methods and pharmaceutical compositions thereof. The present invention also relates to the use of the above-mentioned compound or its pharmaceutical composition in the treatment of SHP2-mediated diseases.
背景技术Background technique
含Src同源区2蛋白质酪氨酸磷酸酶2(Src homologyregion 2-containing protein tyrosine phosphatase 2,SHP2)是由一种由PTPN11基因编码的非受体型蛋白质酪氨酸磷酸酶，PTPN11是首个被发现的编码酪氨酸激酶的原癌基因(Chan R J et al.PTPN11 is the first identified proto-oncogene that encodes a tyrosine phosphatase.Blood,2007,109:862-867)，其编码的SHP2蛋白包含N端的SHP2结构域(N-SHP2)、C端SHP2结构域(C-SHP2)、蛋白质磷酸酶催化结构域(PTP)，两个C端的酪氨酸残基(Y542和Y580)以及一个富含脯氨酸(Pro)的模体。Src homology region 2 protein tyrosine phosphatase 2 (Src homology region 2-containing protein tyrosine phosphatase 2, SHP2) is a non-receptor protein tyrosine phosphatase encoded by the PTPN11 gene, and PTPN11 is the first The discovered proto-oncogene encoding tyrosine kinase (Chan R J et al.PTPN11 is the first identified proto-oncogene that encodes a tyrosine phosphatase.Blood, 2007,109:862-867), and the encoded SHP2 protein contains N-terminal SHP2 domain (N-SHP2), C-terminal SHP2 domain (C-SHP2), protein phosphatase catalytic domain (PTP), two C-terminal tyrosine residues (Y542 and Y580) and one rich Proline (Pro) motif.
近年研究主要认为Ras/ERK通路是SHP2发挥作用最重要的一条信号转导通路，其机制(Dance M et al.The molecular functions of Shp2 in the RAS/mitogen-activated protein kinase(ERK1/2)pathway.Cell Signal,2008,20:453-459)大致为：生长因子受体活化后，其酪氨酸残基发生自体磷酸化，为Grb2和SHP2(含有SH2结构域的衔接蛋白)磷酸酪氨酸结合区域SH2提供停靠位点。Grb2与磷酸化的生长因子受体的结合导致SOS蛋白在胞膜的聚集。SOS作为一种鸟嘌呤核苷酸交换因子(guanine nucleotide exchange factor,GEF)，可以催化膜结合蛋白Ras从无活性的Ras-GDP转换为有活性的Ras-GTP。Ras-GTP再进一步与下游的信号***发生联系，激活Ser/Thr激酶Raf1等，进而在调节激酶MEK的作用下使ERK活化，ERK活化后直接作用于细胞质的靶分子或转移到细胞核内调节基因转录，使细胞增殖或分化。这一过程可能还受到SHP2结合蛋白和底物(SHP substrate-1,SHPS-1)、Ras-GTP酶活化蛋白(Ras-GAP)以及其他Src成员的影响。In recent years, research mainly believes that the Ras/ERK pathway is the most important signal transduction pathway for SHP2, and its mechanism (Dance M et al. The molecular functions of Shp2 in the RAS/mitogen-activated protein kinase (ERK1/2) pathway. Cell Signal, 2008, 20:453-459) roughly: After the growth factor receptor is activated, its tyrosine residues undergo autophosphorylation, which is the combination of Grb2 and SHP2 (adapter protein containing SH2 domain) phosphotyrosine Area SH2 provides stop points. The binding of Grb2 to phosphorylated growth factor receptors results in the accumulation of SOS protein in the cell membrane. SOS, as a guanine nucleotide exchange factor (GEF), can catalyze the conversion of membrane-bound protein Ras from inactive Ras-GDP to active Ras-GTP. Ras-GTP further connects with downstream signaling systems to activate Ser/Thr kinase Raf1, etc., and then activate ERK under the action of the regulatory kinase MEK. After activation of ERK, it directly acts on target molecules in the cytoplasm or transfers to the nucleus to regulate genes Transcription to make cells proliferate or differentiate. This process may also be affected by SHP2 binding protein and substrate (SHP substrate-1, SHPS-1), Ras-GTPase activating protein (Ras-GAP) and other members of Src.
SHP2蛋白不仅调节Ras/ERK信号通路，另有报道其还调节JAK-STAT3、NF-κB、PI3K/Akt、RHO和NFAT等多条信号通路，进而调节细胞增殖、分化、迁移、凋亡等生理学功能。SHP2 protein not only regulates the Ras/ERK signaling pathway, it is also reported to regulate multiple signaling pathways such as JAK-STAT3, NF-κB, PI3K/Akt, RHO, and NFAT, thereby regulating cell proliferation, differentiation, migration, and apoptosis Features.
SHP2被证明与多种疾病相关，Tartaglia等(Tartaglia M et al.Mutations in PTPN11,encoding the protein tyrosine phosphatase SHP-2,cause Noonan syndrome.Nat Genet,2001,29:465-468)发现大约50％的努南综合征患者伴有PTPN11的错义突变。另外，研究发现PTPN11突变是JMML以及多种白血病发病的重要原因(Tartaglia M et al.Nat Genet,2003,34:148-150；Loh ML et al.Blood,2004,103:2325-2331；Tartaglia M et al.Br J Haematol,2005,129:333-339；Xu R et al.Blood,2005,106:3142-3149.)。随着对PTPN11/SHP2研究的深入，发现其与肺癌、胃癌、结肠癌、黑色素瘤、甲状腺癌等多种癌症的发生均有的关系(唐春兰等.中国肺癌杂志，2010,13:98-101；Higuchi M et al.Cancer Sci,2004,95:442-447；Bentires-Al j M et al.Cancer Res,2004,64:8816-8820；Martinelli S et al.Cancer Genet Cytogenet,2006,166:124-129.)。SHP2 has been proved to be related to many diseases, Tartaglia et al. (Tartaglia M et al. Mutations in PTPN11, encoding the protein tyrosine phosphatase SHP-2, cause Noonan syndrome. Nat Genet, 2001, 29:465-468) found that about 50% Patients with Noonan syndrome have missense mutations of PTPN11. In addition, studies have found that PTPN11 mutation is an important cause of JMML and a variety of leukemias (Tartaglia M et al. Nat Genet, 2003, 34: 148-150; Loh ML et al. Blood, 2004, 103: 2325-2331; Tartaglia M et al. Br J Haematol, 2005, 129: 333-339; Xu R et al. Blood, 2005, 106: 3142-3149.). With the deepening of research on PTPN11/SHP2, it has been found that it is related to the occurrence of lung cancer, gastric cancer, colon cancer, melanoma, thyroid cancer and other cancers (Tang Chunlan et al. Chinese Journal of Lung Cancer, 2010, 13: 98- 101; Higuchi M et al. Cancer Sci, 2004, 95: 442-447; Bentiers-Al j M et al. Cancer Res, 2004, 64: 8816-8820; Martinelli S et al. Cancer Genet Cytogenet, 2006, 166: 124-129.).
因此，SHP2抑制剂作为潜在的治疗手段得到了越来越多的关注。目前在开发的SHP2抑制剂有多种，诺华开发的TNO155在2017年进入治疗实体瘤的I期临床试验。加科思设计开发的JAB-3068于2018年1月正式获得美国FDA新药临床实验许可。Revolution开发的RMC-4630于2018年下半年进行首次人体临床试验。目前，该靶点在国内外还未见上市品种，因此，开发出能够靶向抑制SHP2活性的小分子药物，为患者提供更加安全有效的SHP2抑制剂具有重要的研究意义。Therefore, SHP2 inhibitors have received more and more attention as potential treatments. There are a variety of SHP2 inhibitors currently under development. Novartis's TNO155 entered phase I clinical trials for the treatment of solid tumors in 2017. JAB-3068, designed and developed by Jacos, was officially approved by the US FDA for clinical trials of new drugs in January 2018. The RMC-4630 developed by Revolution undergoes its first human clinical trial in the second half of 2018. At present, there is no marketed product for this target at home and abroad. Therefore, the development of small molecule drugs that can target and inhibit the activity of SHP2 and provide patients with safer and more effective SHP2 inhibitors have important research significance.
发明内容Summary of the invention
本发明涉及一种作为含Src同源区2蛋白质酪氨酸磷酸酶2(SHP2)抑制剂应用的化合物。本发明所述化合物具有如式I所示通式结构或其药学上可接受的盐、互变异构体、溶剂化物、螯合物、非共价复合物或前体药物，The present invention relates to a compound used as an inhibitor of protein tyrosine phosphatase 2 (SHP2) containing Src homology region 2. The compound of the present invention has a general structure as shown in Formula I or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,
Figure PCTCN2020072774-appb-000001
Figure PCTCN2020072774-appb-000001
其中，among them,
R 1任意地选自氨基、-C(O)-R a、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基； Optionally R 1 is selected from amino, -C (O) -R a, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, C 1-8 alkoxy containing substituents;
R a任意地选自氨基、-NH-OH、C 1-3烷基； R a is optionally selected from amino, -NH-OH, C 1-3 alkyl;
R 2任意地选自氢、C 1-4烷基或含取代基的C 1-4烷基； Optionally R 2 is selected from hydrogen, C 1-4 alkyl group or a substituted C 1-4 alkyl;
R 3任意地选自氢、卤素、氨基、-C(O)NH 2、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基或含取代基的C 1-8烷氧基； Optionally R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
R 4任意地选自氢、卤素、氨基、酰胺基、-C≡N、羧基、羟基、羟甲基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基或含取代基的C 2-8炔基； Optionally R 4 is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, hydroxymethyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl base;
A 1任意地选自CR 5或N； A 1 is arbitrarily selected from CR 5 or N;
A 2任意地选自CR 6或N； A 2 is arbitrarily selected from CR 6 or N;
A 3任意地选自CR 7或N； A 3 is arbitrarily selected from CR 7 or N;
A 4任意地选自CR 8或N； A 4 is arbitrarily selected from CR 8 or N;
U任意地选自C(R 9) 2、O或NR 10； U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
其中，R 5、R 6、R 7、R 8、R 9或R 10独立地选自氢、羟基、卤素、氨基、含取代基的氨基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基、含取代基的C 2-8炔基或C 5-6杂环基；或者， Wherein, R 5 , R 6 , R 7 , R 8 , R 9 or R 10 are independently selected from hydrogen, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted C 1 -8 alkyl, C 1-8 alkoxy, substituted C 1-8 alkoxy, C 2-8 alkenyl, substituted C 2-8 alkenyl, C 2-8 alkynyl, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents; or,
R 5和R 6与他们连接的碳原子共同形成5到6元芳基或5到6元杂环基； R 5 and R 6 and the carbon atom to which they are attached together form a 5- to 6-membered aryl group or a 5- to 6-membered heterocyclic group;
L选自S；L is selected from S;
环A任意地选自C 6-10芳基或C 5-10杂芳基，所述C 5-10杂芳基含有一个或两个N或S杂原子； Ring A is optionally selected from a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group contains one or two N or S heteroatoms;
Rx任意地选自氢、羟基、卤素、氰基、氨基、含取代基的氨基、磺酰基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 3-8环烷基或含取代基的C 3-8环烷基； Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;
n为0、1、2、3或4。n is 0, 1, 2, 3, or 4.
一些实施方式中，式I中的R 1任意地选自氨基、-C(O)NH 2、-C≡N、C 1-3烷基、含取代基的C 1-3烷基或C 1-3烷氧基。 In some embodiments, R 1 in Formula I optionally selected from amino, -C (O) NH 2, -C≡N, C 1-3 alkyl group, a substituted C 1-3 alkyl group or a C 1 -3 Alkoxy.
一些实施方式中，式I中的R 1任意地选自氨基、-C(O)NH 2、-C≡N、羟基取代的甲基、
Figure PCTCN2020072774-appb-000002
In some embodiments, R 1 in Formula I is arbitrarily selected from amino, -C(O)NH 2 , -C≡N, methyl substituted by hydroxy,
Figure PCTCN2020072774-appb-000002
一些实施方式中，式I中的R 2为氢。 In some embodiments, R 2 in Formula I is hydrogen.
一些实施方式中，式I中的R 3任意地选自氢、卤素、C 1-3烷基或含取代基的C 1-3烷基。 In some embodiments, R 3 of Formula I is optionally selected from hydrogen, halo, C 1-3 alkyl or an optionally substituted C 1-3 alkyl.
一些实施方式中，式I中的R 3任意地选自氢、氯或甲基。 In some embodiments, R 3 in Formula I is arbitrarily selected from hydrogen, chlorine, or methyl.
一些实施方式中，式I中的R 4任意地选自氢、卤素、含取代基的C 1-8烷基或C 1-8烷氧基。 In some embodiments, R 4 in Formula I is arbitrarily selected from hydrogen, halogen, C 1-8 alkyl or C 1-8 alkoxy containing substituents.
一些实施方式中，式I中的R 4任意地选自氢、F、Cl、-CHF 2、CF 3或-O-CH 3。 In some embodiments, R 4 in Formula I is arbitrarily selected from hydrogen, F, Cl, -CHF 2 , CF 3 or -O-CH 3 .
一些实施方式中，式I中的A 1为CR 5或N，其中R 5选自氢、卤素或卤素取代的C 1-3烷基。 In some embodiments, A 1 in formula I is CR 5 or N, wherein R 5 is selected from hydrogen, halogen, or halogen substituted C 1-3 alkyl.
一些实施方式中，式I中的A 1为CR 5或N，其中R 5选自氢、Cl或三氟甲基。 In some embodiments, A 1 in formula I is CR 5 or N, wherein R 5 is selected from hydrogen, Cl or trifluoromethyl.
一些实施方式中，式I中的A 2为CR 6或N，其中R 6选自氢、羟基、卤素、氨基或C 1-8烷氧基。 In some embodiments, A 2 in Formula I is CR 6 or N, wherein R 6 is selected from hydrogen, hydroxyl, halogen, amino, or C 1-8 alkoxy.
一些实施方式中，式I中的A 2为CR 6或N，其中R 6选自氢、OH、F、Cl、氨基或-O-CH 3。 In some embodiments, A 2 in Formula I is CR 6 or N, wherein R 6 is selected from hydrogen, OH, F, Cl, amino, or -O-CH 3 .
一些实施方式中，式I中的A 3为CR 7或N，其中R 7为氢或卤素。 In some embodiments, A 3 in formula I is CR 7 or N, wherein R 7 is hydrogen or halogen.
一些实施方式中，式I中的A 4为CR 8或N，其中R 8选自氢、卤素、氨基、取代氨基、C 1-3烷氧基或C 5-6杂环基。 In some embodiments, A 4 in formula I is CR 8 or N, wherein R 8 is selected from hydrogen, halogen, amino, substituted amino, C 1-3 alkoxy, or C 5-6 heterocyclyl.
一些实施方式中，式I中的A 4为CR 8或N，其中R 8选自氢、F、Cl、氨基、-NHCH 3、-N(CH 3) 2、
Figure PCTCN2020072774-appb-000003
或甲氧基。 In some embodiments, A 4 in formula I is CR 8 or N, wherein R 8 is selected from hydrogen, F, Cl, amino, -NHCH 3 , -N(CH 3 ) 2 ,
Figure PCTCN2020072774-appb-000003
Or methoxy.
一些实施方式中，式I中的U为CH 2或O。 In some embodiments, U in Formula I is CH 2 or O.
一些实施方式中，式I中的环A任意地选自苯基或C 5-6杂芳基，所述C 5-6杂芳基含有一个或两个N或S杂原子。 In some embodiments, ring A in formula I is arbitrarily selected from phenyl or C 5-6 heteroaryl, and the C 5-6 heteroaryl contains one or two N or S heteroatoms.
一些实施方式中，式I中的Rx任意地选自氢、羟基、卤素、氰基、C 1-3烷基、卤素取代的C 1-3烷基或C 1-3烷氧基。 In some embodiments, Rx in Formula I is arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C 1-3 alkyl, halogen-substituted C 1-3 alkyl, or C 1-3 alkoxy.
一些实施方式中，式I中的Rx任意地选自氢、OH、F、Cl、Br、-CN、三氟甲基或甲氧基。In some embodiments, Rx in Formula I is arbitrarily selected from hydrogen, OH, F, Cl, Br, -CN, trifluoromethyl, or methoxy.
一些实施方式中，式I中的
Figure PCTCN2020072774-appb-000004
选自
Figure PCTCN2020072774-appb-000005
Figure PCTCN2020072774-appb-000006
In some embodiments, the formula I
Figure PCTCN2020072774-appb-000004
Selected from
Figure PCTCN2020072774-appb-000005
Figure PCTCN2020072774-appb-000006
本发明进一步提供了式I所示化合物的一些优选技术方案。例如，本发明所述化合物具有如式II所示通式结构或其药学上可接受的盐、互变异构体、溶剂化物、螯合物、非共价复合物或前体药物，The present invention further provides some preferred technical solutions of the compound represented by formula I. For example, the compound of the present invention has the general structure shown in Formula II or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,
Figure PCTCN2020072774-appb-000007
Figure PCTCN2020072774-appb-000007
其中，among them,
R 3任意地选自氢、卤素、氨基、-C(O)NH 2、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基或含取代基的C 1-8烷氧基； Optionally R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
A 1任意地选自CR 5或N； A 1 is arbitrarily selected from CR 5 or N;
U任意地选自C(R 9) 2、O或NR 10； U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
其中，R 5、R 9或R 10独立地选自氢、羟基、卤素、氨基、含取代基的氨基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基或含取代基的C 2-8炔基或C 5-6杂环；或者， Wherein, R 5 , R 9 or R 10 are independently selected from hydrogen, hydroxy, halogen, amino, substituted amino, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl Group or C 5-6 heterocycle; or,
L选自S；L is selected from S;
R x任意地选自氢、羟基、卤素、氰基、氨基、含取代基的氨基、磺酰基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 3-8环烷基或含取代基的C 3-8环烷基； R x is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy group, a substituted group containing C 1-8 alkoxy, C 3-8 cycloalkyl or C 3-8 cycloalkyl group having substituent;
n为0、1、2、3或4。n is 0, 1, 2, 3, or 4.
一些实施方式中，式II中的R 3任意地选自氢或C 1-3烷基。 In some embodiments, R 3 in Formula II is optionally selected from hydrogen or C 1-3 alkyl.
一些实施方式中，式II中的R 3任意地选自氢或甲基。 In some embodiments, R 3 in Formula II is arbitrarily selected from hydrogen or methyl.
一些实施方式中，式II中的A 1为CR 5或N，其中R 5选自卤素。 In some embodiments, A 1 in formula II is CR 5 or N, wherein R 5 is selected from halogen.
一些实施方式中，式II中的A 1为CR 5，其中R 5选自F或Cl。 In some embodiments, A 1 in Formula II is CR 5 , wherein R 5 is selected from F or Cl.
一些实施方式中，式II中的A 1为N。 In some embodiments, A 1 in Formula II is N.
一些实施方式中，式II中的U为O或CH 2。 In some embodiments, U in Formula II is O or CH 2 .
一些实施方式中，式II中的R x任意地选自氢、羟基、卤素、氰基、C 1-3烷基、卤素取代的C 1-3烷基或C 1-3烷氧基。 In some embodiments, R x in Formula II is arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C 1-3 alkyl, halogen-substituted C 1-3 alkyl, or C 1-3 alkoxy.
一些实施方式中，式II中的R x任意地选自氢、OH、CN、Cl、F、Br、三氟甲基或甲氧基。 In some embodiments, R x in Formula II is arbitrarily selected from hydrogen, OH, CN, Cl, F, Br, trifluoromethyl, or methoxy.
一些实施方式中，式II中的n为0、1或2。In some embodiments, n in Formula II is 0, 1, or 2.
本发明进一步提供了式I所示化合物的一些优选技术方案。例如，本发明所述化合物具有如式III所示通式结构或其药学上可接受的盐、互变异构体、溶剂化物、螯合物、非共价复合物或前体药物，The present invention further provides some preferred technical solutions of the compound represented by formula I. For example, the compound of the present invention has the general structure shown in Formula III or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,
Figure PCTCN2020072774-appb-000008
Figure PCTCN2020072774-appb-000008
R 3任意地选自氢、卤素、氨基、-C(O)NH 2、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基或含取代基的C 1-8烷氧基； Optionally R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
R 4任意地选自氢、卤素、氨基、酰胺基、-C≡N、羧基、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基或含取代基的C 2-8炔基； Optionally R 4 is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, A substituted C 1-8 alkoxy group, a C 2-8 alkenyl group, a substituted C 2-8 alkenyl group, a C 2-8 alkynyl group or a substituted C 2-8 alkynyl group;
A 1任意地选自CR 5或N； A 1 is arbitrarily selected from CR 5 or N;
A 2任意地选自CR 6或N； A 2 is arbitrarily selected from CR 6 or N;
A 3任意地选自CR 7或N； A 3 is arbitrarily selected from CR 7 or N;
U任意地选自C(R 9) 2、O或NR 10； U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
其中，R 5、R 6、R 7、R 8、R 9或R 10独立地选自氢、羟基、卤素、氨基、含取代基的氨基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基、含取代基的C 2-8炔基或C 5-6杂环基； Wherein, R 5 , R 6 , R 7 , R 8 , R 9 or R 10 are independently selected from hydrogen, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted C 1 -8 alkyl, C 1-8 alkoxy, substituted C 1-8 alkoxy, C 2-8 alkenyl, substituted C 2-8 alkenyl, C 2-8 alkynyl, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents;
L选自S；L is selected from S;
环A任意地选自C 6-10芳基或C 5-10杂芳基，所述C 5-10杂芳基含有一个或两个N或S杂原子； Ring A is optionally selected from a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group contains one or two N or S heteroatoms;
Rx任意地选自氢、羟基、卤素、氰基、氨基、含取代基的氨基、磺酰基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 3-8环烷基或含取代基的C 3-8环烷基； Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;
n为0、1、2、3或4。n is 0, 1, 2, 3, or 4.
一些实施方式中，式III中的R 3选自氢。 In some embodiments, R 3 in Formula III is selected from hydrogen.
一些实施方式中，式III中的R 4任意地选自氢或Cl。 In some embodiments, R 4 in Formula III is arbitrarily selected from hydrogen or Cl.
一些实施方式中，式III中的A 1为CR 5或N，其中R 5选自氢、Cl或三氟甲基。 In some embodiments, A 1 in Formula III is CR 5 or N, wherein R 5 is selected from hydrogen, Cl or trifluoromethyl.
一些实施方式中，式III中的A 2为CR 6或N，其中R 6选自Cl、氨基、
Figure PCTCN2020072774-appb-000009
或-O-CH 3。 In some embodiments, A 2 in formula III is CR 6 or N, wherein R 6 is selected from Cl, amino,
Figure PCTCN2020072774-appb-000009
Or -O-CH 3 .
一些实施方式中，式III中的A 3为CR 7或N，其中R 7为氢。 In some embodiments, A 3 in Formula III is CR 7 or N, wherein R 7 is hydrogen.
一些实施方式中，式III中的U为CH 2。 In some embodiments, U in Formula III is CH 2 .
一些实施方式中，式III中的环A任意地选自苯基或C 5-6杂芳基，所述C 5-6杂芳基含有一个或两个N或S杂原子。 In some embodiments, ring A in formula III is arbitrarily selected from phenyl or C 5-6 heteroaryl, and the C 5-6 heteroaryl contains one or two N or S heteroatoms.
一些实施方式中，式III中的Rx任意地选自氢、Cl、Br、三氟甲基或甲氧基。In some embodiments, Rx in Formula III is arbitrarily selected from hydrogen, Cl, Br, trifluoromethyl, or methoxy.
一些实施方式中，式III中的
Figure PCTCN2020072774-appb-000010
选自
Figure PCTCN2020072774-appb-000011
Figure PCTCN2020072774-appb-000012
In some embodiments, the formula III
Figure PCTCN2020072774-appb-000010
Selected from
Figure PCTCN2020072774-appb-000011
Figure PCTCN2020072774-appb-000012
本发明进一步提供了一种化合物或其药学上可接受的盐，其中，所述化合物选自：The present invention further provides a compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from:
2)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫代)-5-甲基吡嗪-2-基)甲醇；2) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)-5-methylpyrazin-2-yl)methanol;
3)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲腈；3) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine) -4-yl)thio)pyrazine-2-carbonitrile;
4)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫基)吡嗪-2-甲酰胺；4)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorophenyl )Thio)pyrazine-2-carboxamide;
5)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲酰胺；5) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine) -4-yl)thio)pyrazine-2-carboxamide;
7)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-甲氧基吡啶-4-基)硫基)吡嗪-2-甲酰胺；7) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methoxy (Pyridin-4-yl)thio)pyrazine-2-carboxamide;
8)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-甲酰胺；8)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl) Pyridin-3-yl)thio)pyrazine-2-carboxamide;
9)(S)-3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-甲酰胺；9)(S)-3-(1-Amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2- (Trifluoromethyl)pyridin-3-yl)thio)pyrazine-2-carboxamide;
10)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-溴-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；10) (S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-bromo-1,3-dihydrospiro(indene-2, 4'-Piperidine]-1'-yl)pyrazine-2-carboxamide;
11)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；11) (S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-methoxy-1,3-dihydrospiro(indene- 2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
12)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；12) (S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-methoxy-1,3-dihydrospiro(indene- 2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
13)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-5,6-二甲基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；13)(S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro[ Indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
14)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；14)(S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-(trifluoromethyl)-1,3-dihydrospiro [Indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
15)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氯-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；15) (S)-6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2, 4'-Piperidine]-1'-yl)pyrazine-2-carboxamide;
16)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((6-氨基-3-氯吡啶-2-基)硫基)吡嗪-2-甲酰胺；16)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((6-amino-3-chloropyridine -2-yl)thio)pyrazine-2-carboxamide;
17)(S)-3-(5-氨基-2-甲氧基-5,7-二氢螺[环戊二烯[b]吡啶-6,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲酰胺；17)(S)-3-(5-Amino-2-methoxy-5,7-dihydrospiro[cyclopentadiene[b]pyridine-6,4'-piperidine]-1'-yl) -6-((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide;
18)(S)-3-(4-氨基-2-氯-4,6-二氢螺[环戊二烯并[d]噻唑-5,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲酰胺；18)(S)-3-(4-Amino-2-chloro-4,6-dihydrospiro[cyclopenta[d]thiazole-5,4'-piperidine]-1'-yl)- 6-((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide;
19)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氟-3-甲氧基苯基)硫基)吡嗪-2-基)甲醇；19)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-fluoro-3-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;
20)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-甲氧基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；20)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-methoxypyrimidine- 4-yl)thio)pyrazin-2-yl)methanol;
21)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)甲醇；21)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloro Pyridin-4-yl)thio)pyrazin-2-yl)methanol;
22)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；22)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-4- (Yl)thio)pyrazin-2-yl)methanol;
23)(S)-1-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)乙-1-酮；23)(S)-1-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)ethan-1-one;
24)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；24)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-( (1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol;
25)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-(三氟甲基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；25)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-(trifluoromethyl )Pyridin-4-yl)thio)pyrazin-2-yl)methanol;
26)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；26)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-(trifluoromethyl)-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
27)1-(3-((S)-1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)乙-1-醇；27) 1-(3-((S)-1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)ethan-1-ol;
28)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((5-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；28)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((5-chloro-2-( (1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol;
29)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(二甲基氨基)-3-氟吡啶-4-基)硫基)吡嗪-2-基)甲醇；29)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(dimethylamino )-3-fluoropyridin-4-yl)thio)pyrazin-2-yl)methanol;
30)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-吡啶]-1'-基)-6-((3-氟-2-(甲基氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；30)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-fluoro-2-(methyl (Amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol;
31)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(二氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲基；31)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(difluoromethyl )Pyridin-3-yl)thio)pyrazin-2-yl)methyl;
32)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-(二甲基氨基)吡啶-4-基)硫基)吡嗪-2-基)甲基；32)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-( Dimethylamino)pyridin-4-yl)thio)pyrazin-2-yl)methyl;
33)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氟-2-甲氧基苯基)硫基)吡嗪-2-基)甲醇；33)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-fluoro-2-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;
34)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-5-氟-2-甲氧基苯基)硫基)吡嗪-2-基)甲醇；34)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-5-fluoro -2-Methoxyphenyl)thio)pyrazin-2-yl)methanol;
35)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(喹啉-4-基硫基)吡嗪-2-基)甲醇；35)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(quinolin-4-ylsulfanyl )Pyrazin-2-yl)methanol;
36)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲醇；36)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl )Pyridin-3-yl)thio)pyrazin-2-yl)methanol;
37)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫基)吡嗪-2-基)甲醇；37)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)pyrazin-2-yl)methanol;
38)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲醇；38)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl )Pyridin-3-yl)thio)pyrazin-2-yl)methanol;
39)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氟吡啶-4-基)硫基)吡嗪-2-基)甲基；39)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-fluoro Pyridin-4-yl)thio)pyrazin-2-yl)methyl;
40)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-甲氧基吡啶-4-基)硫基)吡嗪-2-基)甲醇；40)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methyl Oxypyridin-4-yl)thio)pyrazin-2-yl)methanol;
41)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氟-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；41)(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-fluoro-1,3-dihydrospiro(indene-2 ,4'-Piperidine]-1'-yl)pyrazin-2-yl)methanol;
42)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌 啶]-1'-基)吡嗪-2-基)甲醇；42)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-methoxy-1,3-dihydrospiro(indene -2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
43)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-5,6-二甲基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；43)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro [Indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
44)(R)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(3-氨基-3H-螺[苯并呋喃-2,4'-哌啶]-1'-基)吡嗪-2-基)甲基；44)(R)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(3-amino-3H-spiro[benzofuran-2,4'-piperidine ]-1'-yl)pyrazin-2-yl)methyl;
45)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-4-腈；45)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-4-carbonitrile;
46)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氯-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；46)(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro(indene-2 ,4'-Piperidine]-1'-yl)pyrazin-2-yl)methanol;
47)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(吡啶-4-基硫基)吡嗪-2-基)甲醇；47)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(pyridin-4-ylsulfanyl) Pyrazin-2-yl)methanol;
48)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(5-氨基-5,7-二氢螺[环戊烯并[b]吡啶-6,4'-哌啶]-1'-基)吡嗪-2-基)甲基；48)(S)-(6-((2-amino-3-chloropyridin-4-yl)thio)-3-(5-amino-5,7-dihydrospiro[cyclopenteno[b] Pyridine-6,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl;
49)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；49)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-(trifluoromethyl)-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
50)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-6-醇；50)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-6-ol;
51)(S)-4-((5-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(羟基甲基)吡嗪-2-基)硫基)嘧啶-2-醇；51)(S)-4-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyridine (Azin-2-yl)thio)pyrimidin-2-ol;
52)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-7-氟-1,3-二氢螺[茚-2,4'-哌啶]-4-醇；52)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 7-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-4-ol;
53)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氢-[1,4]二氧杂环己并[2,3-b]吡啶-8-基)硫基)吡嗪-2-基)甲醇；53)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dihydro- [1,4]Dioxo[2,3-b]pyridin-8-yl)thio)pyrazin-2-yl)methanol;
54)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-6-腈；54)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-6-nitrile;
55)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-4-氟-1,3-二氢螺[茚-2,4'-哌啶]-7-腈；55)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 4-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-7-nitrile;
56)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-7-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲基；56)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-7-(trifluoromethyl)-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl;
57)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-7-醇；57)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-7-ol;
58)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氟吡啶-4-基)硫基)吡嗪-2-甲酰胺；58)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-fluoropyridine -4-yl)thio)pyrazine-2-carboxamide;
59)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-甲酰胺；59)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-(( 1-Methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazine-2-carboxamide;
60)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)-N-羟基吡嗪-2-甲酰胺；60)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine -4-yl)thio)-N-hydroxypyrazine-2-carboxamide;
61)1-(3-((S)-1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)-2,2,2-三氟乙-1-醇；61)1-(3-((S)-1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)-2,2,2-trifluoroethane-1-ol;
62)(S)-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(甲氧基甲基l)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-1-胺；62)(S)-1'-(5-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(methoxymethyll)pyrazin-2-yl)-1 ,3-Dihydrospiro[indene-2,4'-piperidine]-1-amine;
63)(S)-2-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)丙-2-醇；63)(S)-2-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)propan-2-ol;
64)(S)-(3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；64)(S)-(3-(1-amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2 -Aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol;
65)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(嘧啶-4-基硫基)吡嗪-2-基)甲醇；65)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(pyrimidin-4-ylthio) Pyrazin-2-yl)methanol;
66)(S)-(3-(1-氨基-5-氯-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；66)(S)-(3-(1-Amino-5-chloro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;
67)(S)-(3-(1-氨基-5-氟-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；67)(S)-(3-(1-amino-5-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;
68)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(甲基氨基)嘧啶-4-基)硫基)吡嗪-2-基)甲醇；68)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(methylamino) Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;
69)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)-5-甲基哌嗪-2-基)甲醇；或69)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-4- Yl)thio)-5-methylpiperazin-2-yl)methanol; or
70)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-甲酰胺。70)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((2-aminopyrimidin-4-yl ) Thio) pyrazine-2-carboxamide.
本发明还提供了一种药物组合物，其特征在于，包含治疗有效量的至少一种式(I)所示的化合物和至少一种药学上可接受的辅料。The present invention also provides a pharmaceutical composition characterized by comprising a therapeutically effective amount of at least one compound represented by formula (I) and at least one pharmaceutically acceptable excipient.
本发明进一步提供了一种药物组合物，其特征在于，治疗有效量的至少一种式(I)所示的化合物和药学上可接受的辅料的质量百分比为0.0001:1-10。The present invention further provides a pharmaceutical composition, characterized in that the mass percentage of the therapeutically effective amount of at least one compound represented by formula (I) and pharmaceutically acceptable excipients is 0.0001:1-10.
本发明提供了结构式(I)所示化合物或药物组合物在制备药物中的应用。The invention provides the application of the compound or pharmaceutical composition represented by the structural formula (I) in the preparation of medicines.
本发明进一步提供了所述应用的优选技术方案：The present invention further provides a preferred technical solution for the application:
作为优选，所述应用为制备用于治疗、预防、延迟或阻止癌症，癌症转移，心血管疾病，免疫疾病，纤维化或眼部疾病的药物的应用。Preferably, the application is an application for preparing a medicine for treating, preventing, delaying or preventing cancer, cancer metastasis, cardiovascular disease, immune disease, fibrosis or eye disease.
作为优选，所述应用为制备治疗由SHP2介导的疾病的药物中的应用。作为优选，所述疾病是癌症。Preferably, the application is an application in preparing a medicine for treating diseases mediated by SHP2. Preferably, the disease is cancer.
作为优选，所述癌症选自Noonan综合征、豹斑综合征、青少年髓单核细胞白血病、神经母细胞瘤、黑色素瘤、头颈部鳞状细胞癌、急性髓性白血病、乳腺癌、食道肿瘤、肺癌、结肠癌、头癌、胃癌、淋巴瘤、胶质母细胞瘤、胰腺癌或其组合。Preferably, the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal tumor , Lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma, pancreatic cancer or a combination thereof.
作为优选，所述应用为用于制备SHP2抑制剂的应用。Preferably, the application is an application for preparing an SHP2 inhibitor.
本发明还提供了一种治疗和/或预防由SHP2介导的疾病的方法，包括向治疗对象施用治疗有效量的至少任意一种结构式(I)所示化合物或药物组合物。The present invention also provides a method for treating and/or preventing diseases mediated by SHP2, which comprises administering a therapeutically effective amount of at least any one compound or pharmaceutical composition represented by structural formula (I) to a subject.
作为优选，在上述方法中，所述SHP2介导的疾病是癌症。Preferably, in the above method, the SHP2-mediated disease is cancer.
本发明还提供了一种治疗癌症的方法，包括向治疗对象施用治疗有效量的至少任意一种结构式(I)所示化合物或药物组合物。The present invention also provides a method for treating cancer, which comprises administering a therapeutically effective amount of at least any one compound or pharmaceutical composition represented by structural formula (I) to a subject.
作为优选，所述癌症选自Noonan综合征、豹斑综合征、青少年髓单核细胞白血病、神经母细胞瘤、黑色素瘤、头颈部鳞状细胞癌、急性髓性白血病、乳腺癌、食道肿瘤、肺癌、结肠癌、头癌、胃癌、淋巴瘤、胶质母细胞瘤、胰腺癌或其组合。Preferably, the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal tumor , Lung cancer, colon cancer, head cancer, stomach cancer, lymphoma, glioblastoma, pancreatic cancer or a combination thereof.
作为优选，在上述方法中，所述治疗对象为人类。Preferably, in the above method, the treatment target is a human.
除非另有说明，本发明所用术语含义如下：Unless otherwise specified, the terms used in the present invention have the following meanings:
术语“烷基”包括直连、支链或环状的饱和烷基。例如，烷基包括但不限于甲基、乙基、丙基、异丙基、环丙基、正丁基、异丁基、仲丁基、叔丁基、环丁基、正戊基、3-(2-甲基)丁基、2-戊基、2-甲基丁基、新戊基、环戊基、n-己基、2-己基、2-甲基戊基及环己基等类似基团。类似的，“C 1-8烷基”中的“C 1-8”是指包含有1、2、3、4、5、6、7或8个碳原子的直链、支链或环状形式排列的基团。 The term "alkyl" includes straight, branched or cyclic saturated alkyl groups. For example, alkyl includes but is not limited to methyl, ethyl, propyl, isopropyl, cyclopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, cyclobutyl, n-pentyl, 3 -(2-Methyl)butyl, 2-pentyl, 2-methylbutyl, neopentyl, cyclopentyl, n-hexyl, 2-hexyl, 2-methylpentyl and cyclohexyl and other similar groups group. Similar, "C 1-8 alkyl""C1-8" means comprising 7 or 8 carbon atoms, a straight chain, branched chain or cyclic Formally arranged groups.
“烯基”和“炔基”包括直链、支链或环状的烯基和炔基。同样地，“C 2-8烯基”和“C 2-8炔基”是指含有2、3、4、5、6、7或8个碳原子以直链、支链或环状形式排列的烯基或炔基。 "Alkenyl" and "alkynyl" include linear, branched or cyclic alkenyl and alkynyl groups. Similarly, "C 2-8 alkenyl" and "C 2-8 alkynyl" refer to a linear, branched or cyclic arrangement containing 2, 3, 4, 5, 6, 7 or 8 carbon atoms的alkenyl or alkynyl.
术语“烷氧基”是指前述的直链、支链或环状烷基的氧醚形式。The term "alkoxy" refers to the oxyether form of the aforementioned linear, branched or cyclic alkyl group.
术语“芳基”是指未取代或取代的包括碳原子的单环或多环芳香基团。优选为6到10元的单环或双环芳香基团。优选为苯基、萘基。最优选为苯基。The term "aryl" refers to an unsubstituted or substituted monocyclic or polycyclic aromatic group including carbon atoms. It is preferably a 6 to 10 membered monocyclic or bicyclic aromatic group. Preferably it is phenyl and naphthyl. Most preferred is phenyl.
术语“杂芳基”是指，从一个母体杂芳环***的一个碳原子上移走一个氢原子所形成的 单价的杂原子基团。杂芳基包括：5-到7-元芳香、单环，包括至少一个选自N、O或S的杂原子，例如，1到4个杂原子，或优选为1到3个杂原子，环上的其他原子为碳；多杂芳基环包括至少一个选自N、O或S的杂原子，例如，1到4个杂原子，或优选为1到3个杂原子，环上的其他原子为碳，且其中至少一个杂原子在芳环上。特别优选的杂芳基基团是C 3-10的杂芳基，包括但不限于，吡咯基、呋喃基、噻吩基、吡啶基、吡喃基、吡唑基、嘧啶基、哒嗪基、吡嗪基、咪唑基、噻唑基、恶唑基、异恶唑基、三氮唑基、吲哚基、苯并呋喃基、苯并噻唑基、苯并咪唑基、苯并吡唑基、苯并三氮唑基、咔唑基、喹啉基、异喹啉基、嘌呤基等类似基团。 The term "heteroaryl" refers to a monovalent heteroatom group formed by removing a hydrogen atom from a carbon atom of a parent heteroaromatic ring system. Heteroaryl groups include: 5- to 7-membered aromatic, monocyclic, including at least one heteroatom selected from N, O or S, for example, 1 to 4 heteroatoms, or preferably 1 to 3 heteroatoms, ring The other atoms on the above are carbon; the polyheteroaryl ring includes at least one heteroatom selected from N, O or S, for example, 1 to 4 heteroatoms, or preferably 1 to 3 heteroatoms, other atoms in the ring Is carbon, and at least one of the heteroatoms is on the aromatic ring. Particularly preferred heteroaryl groups are C 3-10 heteroaryl groups, including but not limited to, pyrrolyl, furyl, thienyl, pyridyl, pyranyl, pyrazolyl, pyrimidinyl, pyridazinyl, Pyrazinyl, imidazolyl, thiazolyl, oxazolyl, isoxazolyl, triazole, indolyl, benzofuranyl, benzothiazolyl, benzimidazolyl, benzopyrazolyl, benzene And similar groups such as triazolyl, carbazolyl, quinolinyl, isoquinolinyl, purinyl and the like.
但是，在任何情况下，杂芳基和芳基都不会彼此交叉或相互包含。因此，根据以上定义，如果至少一个全碳芳香环与一个杂环基相稠合，得到的是杂芳基，而不是芳基。However, in any case, the heteroaryl group and the aryl group will not cross or contain each other. Therefore, according to the above definition, if at least one all-carbon aromatic ring is fused with a heterocyclic group, the result is a heteroaryl group instead of an aryl group.
“环烷基”指饱和的或不饱和的但不具有芳香性的环状基团。根据其饱和度的特殊水平，分别采用术语“环烷基”、“环烯基”或“环炔基”。有代表性的环烷基基团包括但不限于，环丙烷、环丁烷、环戊烷、环己烷或环己烯等类似基团。具体的，环烷基基团可以是C 3-10的环烷基，如：C 3-6环烷基。 "Cycloalkyl" refers to a cyclic group that is saturated or unsaturated but not aromatic. Depending on the specific level of saturation, the terms "cycloalkyl", "cycloalkenyl" or "cycloalkynyl" are used respectively. Representative cycloalkyl groups include, but are not limited to, cyclopropane, cyclobutane, cyclopentane, cyclohexane, or cyclohexene and similar groups. Specifically, the cycloalkyl group may be a C 3-10 cycloalkyl group, such as a C 3-6 cycloalkyl group.
“杂环基”是指饱和的或不饱和的但不具有芳香性的环状基团，而且其中一个或多个碳原子(以及所连接的氢原子)可分别被相同的或不相同的杂原子和相应所连接的氢原子所取代。有代表性的取代碳原子的杂原子包括但不限于N、P、O、S和Si。当需要描述特定的饱和度时，分别采用术语“杂环烷基”或“杂环烯基”。具有代表性的杂环基基团包括但不限于环氧化合物、咪唑烷、吗啉、哌嗪、哌啶、吡唑烷、吡咯烷、奎宁环、四氢呋喃或四氢吡喃等类似基团。含取代基的杂环基也包含被至少一个含氧的(＝O)或氧化物(-O-)取代基取代的环***，如：哌啶-氮-氧化物、吗啉基-氮-氧化物、1-氧代-1-硫吗啉基和1-二氧-1-硫吗啉基。"Heterocyclic group" refers to a saturated or unsaturated but not aromatic cyclic group, and one or more of the carbon atoms (and the connected hydrogen atoms) can be respectively the same or different hetero Atom and the corresponding attached hydrogen atom are replaced. Representative heteroatoms that replace carbon atoms include, but are not limited to, N, P, O, S, and Si. When a specific degree of saturation needs to be described, the terms "heterocycloalkyl" or "heterocycloalkenyl" are used respectively. Representative heterocyclic groups include but are not limited to epoxy compounds, imidazolidine, morpholine, piperazine, piperidine, pyrazolidine, pyrrolidine, quinuclidine, tetrahydrofuran or tetrahydropyran and similar groups . Substituent heterocyclic groups also include ring systems substituted with at least one oxygen-containing (=O) or oxide (-O-) substituent, such as piperidine-nitrogen-oxide, morpholinyl-nitrogen- Oxide, 1-oxo-1-thiomorpholinyl and 1-dioxo-1-thiomorpholinyl.
但是，在任何情况下，杂环烷基和环烷基都不会彼此交叉或相互包含。因此，根据上述定义，如果至少一个全碳环与一个杂环烷基稠合形成一个二-、多-或螺-环，将仍然定义为杂环烷基。However, in any case, the heterocycloalkyl group and the cycloalkyl group will not cross or contain each other. Therefore, according to the above definition, if at least one all-carbocyclic ring is fused with a heterocycloalkyl to form a di-, poly- or spiro-ring, it will still be defined as a heterocycloalkyl.
另外，如果一个杂芳基与一个杂环基稠和形成一个二-、多-或螺-环，将定义为杂环基而不是杂芳基。In addition, if a heteroaryl group is fused with a heterocyclic group to form a di-, poly- or spiro-ring, it will be defined as a heterocyclic group instead of a heteroaryl group.
“卤素”是指氟(F)、氯(Cl)、溴(Br)或碘(I)。优选的卤素是指氟、氯和溴。"Halogen" refers to fluorine (F), chlorine (Cl), bromine (Br) or iodine (I). Preferred halogen refers to fluorine, chlorine and bromine.
“卤代基”是指氟代、氯代、溴代或碘代基团。优选的卤代基是指氟代和氯代。"Halo" refers to a fluoro, chloro, bromo or iodo group. The preferred halo groups refer to fluoro and chloro.
“取代”是指一个基团中的一个或多个氢原子分别被相同的或不同的取代基所取代。具 有代表性的取代基包括但不限于卤素、氨基、羟基、氧代基、羰基、氰基、-C(O)NH 2、烷基、烷氧基、芳基、环烷基、杂环基、杂芳基。在一些实施例中，取代基包含但不限于卤素、氨基、羟基、氰基、甲基、-CH 2OH、-C(O)NH 2、-OCH 3、三氟甲基。 "Substitution" means that one or more hydrogen atoms in a group are replaced by the same or different substituents. Representative substituents include, but are not limited to, halogen, amino, hydroxy, oxo, carbonyl, cyano, -C (O) NH 2, alkyl, alkoxy, aryl, cycloalkyl, heterocyclyl , Heteroaryl. In some embodiments, the substituent includes, but is not limited to, halogen, amino, hydroxy, cyano, methyl, -CH 2 OH, -C(O)NH 2 , -OCH 3 , trifluoromethyl.
无论何时，术语“烷基”或“芳基”或者其前缀词根出现在取代基名称中(如芳烷基，或二烷基氨基)，均应按前述的“烷基”和“芳基”定义对取代基进行限定性解释。碳原子的指定数量(如C l-6)将独立的表示在一个烷基部分或在一个更大的取代基中的烷基部分(其中烷基作为前缀词根)中的碳原子的数量。 Whenever the term "alkyl" or "aryl" or its prefixes appear in the name of a substituent (such as aralkyl, or dialkylamino), it shall be the same as the aforementioned "alkyl" and "aryl""The definition defines the substituents. The specified number of carbon atoms (such as C 1-6 ) will independently represent the number of carbon atoms in an alkyl moiety or in an alkyl moiety (where the alkyl group is the prefix stem) in a larger substituent.
本发明所述“化合物”包括式(I)所示的化合物，及其所有药学上可接受的形式。这些药学上可接受的形式包括盐、溶剂化物、非共价复合物、螯合物或其前体药物、或上述所有形式的任意混合物。The "compound" of the present invention includes the compound represented by formula (I), and all pharmaceutically acceptable forms thereof. These pharmaceutically acceptable forms include salts, solvates, non-covalent complexes, chelates or their prodrugs, or any mixture of all the above forms.
所述“药学上可接受的”是指公知的用于动物的，特别是可用于人体的。The "pharmaceutically acceptable" refers to those that are well known for use in animals, especially those that can be used in humans.
本发明中术语“组合物”包括含有特定数量的特定组分的产品，也包括任何由特定数量的特定组分直接或间接得到的产品。因此，包括本发明中的化合物作为活性组分的药物组合物和制备该化合物的方法都是本发明的内容。The term "composition" in the present invention includes a product containing a specific quantity of a specific component, and also includes any product directly or indirectly obtained from a specific quantity of a specific component. Therefore, a pharmaceutical composition including the compound of the present invention as an active ingredient and a method for preparing the compound are the content of the present invention.
“治疗有效量”是指一个化合物施用于治疗主体时治疗并且预防和/或抑制一种疾病、病情、症状、适应症和/或不适的至少一种临床症状时，足以这种疾病、病情、症状、适应症或不适的治疗产生一定效果的剂量。具体的“有效治疗剂量”可以根据化合物，给药途径、患者年龄、患者体重，所治疗的疾病或不适的类型、症状和严重程度等的不同而变化。在任意可能的情况下，一个合适的剂量对那些在本领域的专业人员可以是显而易见的，也可以是用常规实验方法确定的。"Therapeutically effective amount" means that when a compound is administered to a subject to treat and prevent and/or inhibit at least one clinical symptom of a disease, condition, symptom, indication, and/or discomfort, it is sufficient for the disease, condition, A dose that produces a certain effect in the treatment of symptoms, indications or discomfort. The specific "effective therapeutic dose" may vary according to the compound, the route of administration, the age of the patient, the weight of the patient, the type of disease or discomfort to be treated, the symptoms and severity, etc. Whenever possible, an appropriate dose may be obvious to those skilled in the art, or it may be determined by conventional experimental methods.
本发明提供的化合物可以以“药学上可接受的盐”的形式存在。药物应用方面，本发明提供的化合物的盐是指无毒的药学上可接受的盐。药学上可接受的盐的形式包括药学上可接受的酸/阴离子或碱/阳离子盐。药学上可接受的酸/阴离子盐一般以碱性氮与无机酸或有机酸质子化的形式存在。典型的有机或无机酸包括盐酸、氢溴酸、氢碘酸、高氯酸、硫酸、硝酸、磷酸、乙酸、丙酸、乙醇酸、乳酸、琥珀酸、马来酸、富马酸、苹果酸、酒石酸、柠檬酸、α-酮戊二酸、马尿酸、苯甲酸、扁桃酸、甲磺酸、羟乙基磺酸、苯磺酸、草酸、扑酸、2-萘磺酸、对甲苯磺酸、环己胺磺酸、水杨酸、糖精酸或三氟乙酸。药学上可接受的碱/阳离子盐，包括但不限于，铝盐、钙盐、氯普鲁卡因盐、胆碱、二乙醇胺盐、乙二胺盐、锂盐、镁盐、钾盐、钠盐和锌盐。The compounds provided by the present invention may exist in the form of "pharmaceutically acceptable salts". In terms of pharmaceutical applications, the salt of the compound provided by the present invention refers to a non-toxic pharmaceutically acceptable salt. The form of the pharmaceutically acceptable salt includes a pharmaceutically acceptable acid/anion or base/cation salt. Pharmaceutically acceptable acid/anionic salts generally exist in the form of protonation of basic nitrogen with inorganic or organic acids. Typical organic or inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, perchloric acid, sulfuric acid, nitric acid, phosphoric acid, acetic acid, propionic acid, glycolic acid, lactic acid, succinic acid, maleic acid, fumaric acid, malic acid , Tartaric acid, citric acid, α-ketoglutaric acid, hippuric acid, benzoic acid, mandelic acid, methanesulfonic acid, isethionic acid, benzenesulfonic acid, oxalic acid, pamoic acid, 2-naphthalenesulfonic acid, p-toluenesulfonic acid Acid, cyclohexylamine sulfonic acid, salicylic acid, saccharic acid or trifluoroacetic acid. Pharmaceutically acceptable base/cation salts, including, but not limited to, aluminum salt, calcium salt, chloroprocaine salt, choline, diethanolamine salt, ethylenediamine salt, lithium salt, magnesium salt, potassium salt, sodium Salt and zinc salt.
本发明化合物的药物前体包含在本发明的保护范围内。通常，所述药物前体是很容易在体内转化成所需要的化合物的功能性衍生物。因此，本发明提供的治疗方法涉及的术语“给药”包括施用本发明公开的化合物，或虽未明确公开但对主体给药后能够在体内转化为本发明公开的化合物治疗所述的各种疾病。有关选择和制备合适药物前体衍生物的常规方法，已记载在例如《药物前体设计》(Design of Prodrugs,ed.H.Bundgaard,Elsevier,1985)这类书中。The prodrug of the compound of the present invention is included in the protection scope of the present invention. Generally, the prodrug is a functional derivative that is easily converted into a desired compound in vivo. Therefore, the term "administration" involved in the treatment method provided by the present invention includes the administration of the compound disclosed in the present invention, or although it is not clearly disclosed but can be transformed into the compound disclosed in the present invention in vivo after administration to the subject. disease. The conventional methods for selecting and preparing suitable prodrug derivatives have been recorded in books such as "Design of Prodrugs" (Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985).
显然的，一个分子中任何取代基或特定位置的变量的定义，与其他分子中的任何取代基或特定位置的变量的定义是无关的。很容易理解，本发明中的化合物可以根据本学科现有技术选择合适的取代基或取代形式，以提供化学上稳定且容易用本学科现有技术或本发明中所述的方法进行制备合成。Obviously, the definition of any substituent or variable at a specific position in one molecule is irrelevant to the definition of any substituent or variable at a specific position in other molecules. It is easy to understand that the compounds of the present invention can be selected according to the prior art of the subject to select suitable substituents or substitution forms to provide chemically stable and easy preparation and synthesis using the prior art of the subject or the method described in the present invention.
当式(I)所示化合物及其药学上可接受的盐为溶剂化物或多晶型的形式时，本发明包括任何可能的溶剂化物和多晶型。形成溶剂化物的溶剂类型没有特别的限定，只要该溶剂是药理学上可以接受的。例如，水、乙醇、丙醇、丙酮等类似的溶剂都可以采用。When the compound represented by formula (I) and its pharmaceutically acceptable salt are in the form of a solvate or polymorph, the present invention includes any possible solvate and polymorph. The type of solvent that forms the solvate is not particularly limited, as long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone and similar solvents can be used.
术语“药学上可接受的盐”是指从药学上可接受的无毒的碱或酸制备的盐。当本发明提供的化合物是酸时，可以从药学上可接受的无毒的碱，包括无机碱和有机碱，制得其相应的盐。从无机碱衍生的盐包括铝、铵、钙、铜(ic和ous)、铁、亚铁、锂、镁、锰(ic和ous)、钾、钠、锌之类的盐。特别地，优选铵、钙、镁、钾和钠的盐。能够衍生成药学上可接受的盐的无毒有机碱包括伯胺、仲胺和叔胺，也包括环胺及含有取代基的胺，如天然存在的和合成的含取代基的胺。能够成盐的其他药学上可接受的无毒有机碱，包括离子交换树脂以及精氨酸、甜菜碱、咖啡因、胆碱、N',N'-二苄乙烯二胺、二乙胺、2-二乙氨基乙醇、2-二甲胺基乙醇、乙醇胺、乙二胺、N－乙基吗啉、N-乙基哌啶、还原葡萄糖胺、氨基葡萄糖、组氨酸、哈胺、异丙胺、赖氨酸，甲基葡萄糖胺、吗啉、哌嗪、哌啶、多胺树脂、普鲁卡因、嘌呤、可可碱、三乙胺、三甲胺、三丙胺、氨丁三醇等。The term "pharmaceutically acceptable salt" refers to a salt prepared from a pharmaceutically acceptable non-toxic base or acid. When the compound provided by the present invention is an acid, its corresponding salt can be prepared from pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper (ic and ous), iron, ferrous, lithium, magnesium, manganese (ic and ous), potassium, sodium, zinc and the like. In particular, salts of ammonium, calcium, magnesium, potassium, and sodium are preferred. Non-toxic organic bases that can be derivatized into pharmaceutically acceptable salts include primary, secondary and tertiary amines, as well as cyclic amines and amines containing substituents, such as naturally occurring and synthetic amines containing substituents. Other pharmaceutically acceptable non-toxic organic bases capable of forming salts, including ion exchange resins and arginine, betaine, caffeine, choline, N',N'-dibenzylethylene diamine, diethylamine, 2 -Diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, reduced glucosamine, glucosamine, histidine, haamine, isopropylamine , Lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.
当本发明提供的化合物是碱时，可以从药学上可接受的无毒的酸，包括无机酸和有机酸，制得其相应的盐。这样的酸包括，如，醋酸、苯磺酸、苯甲酸、樟脑磺酸、柠檬酸、乙磺酸、羟乙基磺酸、甲酸、富马酸、葡萄糖酸、谷氨酸、氢溴酸、盐酸、乳酸、马来酸、苹果酸、扁桃酸、α-酮戊二酸、马尿酸、甲磺酸、黏酸、硝酸、扑酸、泛酸、磷酸、琥珀酸、硫酸、酒石酸、对甲苯磺酸等。较优地，苹果酸、柠檬酸、氢溴酸、盐酸、甲磺酸、马来酸、磷酸、硫酸和酒石酸。更优地，磷酸、盐酸和苹果酸。由于式(I)所示化合物将 作为药物应用，所以优选使用基本上纯的形式，例如，至少60％纯度，更适当至少75％的纯度，特别适当至少98％的纯度(％是重量比)。When the compound provided by the present invention is a base, the corresponding salt can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic acids and organic acids. Such acids include, for example, acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, isethionic acid, formic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, Hydrochloric acid, lactic acid, maleic acid, malic acid, mandelic acid, α-ketoglutaric acid, hippuric acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonate Acid etc. Preferably, malic acid, citric acid, hydrobromic acid, hydrochloric acid, methanesulfonic acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid. More preferably, phosphoric acid, hydrochloric acid and malic acid. Since the compound represented by formula (I) will be used as a medicine, it is preferable to use a substantially pure form, for example, at least 60% purity, more suitably at least 75% purity, particularly suitably at least 98% purity (% is weight ratio) .
本发明提供的药物组合物包括作为活性组分的式(I)所示化合物(或其药学上可接受的盐)，一种药学上可接受的赋形剂及其他可选的治疗组分或辅料。尽管任何给定的情况下，最适合的活性组分给药方式取决于接受给药的特定的主体、主体性质和病情严重程度，但是本发明的药物组合物包括适于口腔、直肠、局部和肠外(包括皮下给药、肌肉注射、静脉给药)给药的药物组合物。本发明的药物组合物可以方便地以本领域公知的单位剂型存在和药学领域公知的任何制备方法制备。The pharmaceutical composition provided by the present invention includes a compound represented by formula (I) (or a pharmaceutically acceptable salt thereof) as an active component, a pharmaceutically acceptable excipient and other optional therapeutic components or Accessories. Although in any given case, the most suitable way of administering the active ingredient depends on the particular subject to be administered, the nature of the subject and the severity of the disease, the pharmaceutical composition of the present invention includes oral, rectal, topical and Pharmaceutical composition for parenteral (including subcutaneous administration, intramuscular injection, intravenous administration) administration. The pharmaceutical composition of the present invention can be conveniently prepared in a unit dosage form known in the art and prepared by any preparation method known in the pharmaceutical field.
实际上，根据常规的药物混合技术，本发明式(I)所示化合物，或药物前体，或代谢物，或药学上可接受的盐，可以合并用药作为活性组分，与药物载体混合成药物组合物。所述药物载体可以采取各种各样的形式，取决于想采用的给药方式，例如，口服或注射(包括静脉注射)。因此，本发明的药物组合物可以采用适于口服给药的独立单位的形式，如包含预先确定剂量的活性组分的胶囊剂，扁囊剂或片剂。进一步地，本发明的药物组合物可采用粉末、颗粒、溶液、水性悬浮液、非水液体、水包油型乳液，或油包水型乳液形式。另外，除了上述提到的常见的剂型，式(I)所示化合物或其药学上可接受的盐，也可以通过控释的方式和/或输送装置给药。本发明的药物组合物可以采用任何制药学上的方法制备。一般情况下，这种方法包括使活性组分和构成一个或多个必要组分的载体缔合的步骤。一般情况下，所述药物组合物经由活性组分与液体载体或精细分割的固体载体或两者的混合物经过均匀的密切混合制得。另外，该产品可以方便地制备成所需要的外观。In fact, according to conventional drug mixing technology, the compound represented by formula (I) of the present invention, or prodrug, or metabolite, or pharmaceutically acceptable salt, can be used in combination as an active ingredient and mixed with a drug carrier to form Pharmaceutical composition. The pharmaceutical carrier can take various forms, depending on the desired mode of administration, for example, oral or injection (including intravenous injection). Therefore, the pharmaceutical composition of the present invention may take the form of a separate unit suitable for oral administration, such as a capsule, cachet or tablet containing a predetermined dose of the active ingredient. Further, the pharmaceutical composition of the present invention may take the form of powder, granule, solution, aqueous suspension, non-aqueous liquid, oil-in-water emulsion, or water-in-oil emulsion. In addition, in addition to the common dosage forms mentioned above, the compound represented by formula (I) or a pharmaceutically acceptable salt thereof can also be administered through a controlled release method and/or a delivery device. The pharmaceutical composition of the present invention can be prepared by any pharmaceutical method. Generally, this method includes the step of associating the active ingredient with the carrier constituting one or more necessary ingredients. In general, the pharmaceutical composition is prepared by uniformly and intimately mixing the active ingredient with a liquid carrier or a finely divided solid carrier or a mixture of both. In addition, the product can be easily prepared into the desired appearance.
因此，本发明的药物组合物包括药学上可接受的载体和式(I)所示化合物，或其药学上可接受的盐。式(I)所示化合物，或其药学上可接受的盐，与其他一种或多种具有治疗活性联合用药的化合物的也包括在本发明的药物组合物中。Therefore, the pharmaceutical composition of the present invention includes a pharmaceutically acceptable carrier and a compound represented by formula (I), or a pharmaceutically acceptable salt thereof. The compound represented by formula (I), or a pharmaceutically acceptable salt thereof, and one or more other compounds with therapeutic activity in combination are also included in the pharmaceutical composition of the present invention.
本发明采用的药物载体可以是，例如，固体载体、液体载体或气体载体。固体载体的例子，包括，乳糖、石膏粉、蔗糖、滑石粉、明胶、琼脂、果胶、***胶、硬脂酸镁、硬脂酸、甘露醇、山梨醇、微晶纤维素、无机盐类、淀粉、预胶化淀粉、糖粉、糊精等。液体载体的例子包括，糖浆、花生油、橄榄油和水。气体载体的例子包括二氧化碳和氮气。制备药物口服制剂时，可以使用任何方便的制药学上的介质。例如，水、乙二醇、油类、醇类、增味剂、防腐剂、着色剂等可用于口服的液体制剂如悬浮剂、酏剂和溶液剂；而载体，如淀粉类、糖类、微晶纤维素、稀释剂、造粒剂、润滑剂、粘合剂、崩解剂等可用于 口服的固体制剂如散剂、胶囊剂和片剂。考虑到易于施用，口服制剂首选片剂和胶囊。可选地，片剂包衣可使用标准的水制剂或非水制剂技术。The drug carrier used in the present invention can be, for example, a solid carrier, a liquid carrier or a gas carrier. Examples of solid carriers include lactose, gypsum powder, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, stearic acid, mannitol, sorbitol, microcrystalline cellulose, inorganic salts , Starch, pregelatinized starch, powdered sugar, dextrin, etc. Examples of liquid carriers include syrup, peanut oil, olive oil and water. Examples of gas carriers include carbon dioxide and nitrogen. When preparing oral pharmaceutical preparations, any convenient pharmaceutical medium can be used. For example, water, ethylene glycol, oils, alcohols, flavor enhancers, preservatives, coloring agents, etc. can be used for oral liquid preparations such as suspensions, elixirs and solutions; and carriers, such as starches, sugars, Microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrants, etc. can be used in oral solid preparations such as powders, capsules and tablets. In view of ease of administration, tablets and capsules are preferred for oral preparations. Alternatively, standard aqueous or non-aqueous formulation techniques can be used for tablet coating.
含有本发明化合物或药物组合物的片剂可通过，可选地，可以与一种或多种辅助组分或辅药一起混合、压制或成型制备。活性组分以可以自由流动的形式如粉末或颗粒，与润滑剂、惰性稀释剂、表面活性或分散剂混合，在适当的机器中，通过压制可以制得压制片剂。用一种惰性液体稀释剂浸湿粉末状的化合物或药物组合物，然后在适当的机器中，通过成型可以制得模制片。较优地，每个片剂含有大约0.01mg到5g的活性组分，每个扁襄剂或胶囊剂含有大约0.1mg到0.5g的活性组分。例如，拟用于人类口服给药的剂型包含约0.1mg到约0.5g的活性组分，与合适且方便计量的辅助材料复合，该辅助材料约占药物组合物总量的5％至99.99％。单位剂型一般包含约0.1mg到约0.5g的有效组分，典型的是0.1mg、0.2mg、0.5mg、1mg、2mg、2.5mg、5mg、10mg、25mg、50mg、100mg、200mg、300mg、400mg或500mg。Tablets containing the compound or pharmaceutical composition of the present invention can be prepared by, optionally, mixing, compression or molding with one or more auxiliary components or adjuvants. The active ingredient is in a free-flowing form such as powder or granules, mixed with lubricants, inert diluents, surface active or dispersing agents, and compressed in a suitable machine to obtain compressed tablets. The powdered compound or pharmaceutical composition is wetted with an inert liquid diluent, and then molded in a suitable machine to form a molded tablet. Preferably, each tablet contains about 0.01 mg to 5 g of active ingredient, and each cachet or capsule contains about 0.1 mg to 0.5 g of active ingredient. For example, a dosage form intended for oral administration to humans contains about 0.1 mg to about 0.5 g of the active ingredient, compounded with a suitable and convenient metering auxiliary material, which accounts for about 5% to 99.99% of the total pharmaceutical composition . The unit dosage form generally contains about 0.1mg to about 0.5g of effective ingredients, typically 0.1mg, 0.2mg, 0.5mg, 1mg, 2mg, 2.5mg, 5mg, 10mg, 25mg, 50mg, 100mg, 200mg, 300mg, 400mg Or 500mg.
本发明提供的适用于胃肠外给药的药物组合物可将活性组分加入水中制备成水溶液或悬浮液。可以包含适当的表面活性剂如十二烷基硫酸钠、聚山梨酯-80(吐温-80)、聚氧乙烯氢化蓖麻油、泊洛沙姆。在甘油、液态聚乙二醇，及其在油中的混合物，也可以制得分散体系。进一步地，防腐剂也可以，包含在本发明的药物组合物中用于防止有害的微生物生长。The pharmaceutical composition suitable for parenteral administration provided by the present invention can be prepared as an aqueous solution or suspension by adding active components into water. Suitable surfactants such as sodium lauryl sulfate, polysorbate-80 (Tween-80), polyoxyethylene hydrogenated castor oil, and poloxamer may be included. In glycerol, liquid polyethylene glycol, and their mixtures in oil, dispersion systems can also be prepared. Furthermore, a preservative may also be included in the pharmaceutical composition of the present invention to prevent the growth of harmful microorganisms.
本发明提供适用于注射使用的药物组合物，包括无菌水溶液或分散体系。进一步地，上述药物组合物可以制备成可用于即时配制无菌注射液的无菌粉末的形式。无论如何，最终的注射形式必须是无菌的，且为了易于注射，必须是易于流动的。此外，所述药物组合物在制备和储存过程中必须稳定。因此，优选抗微生物如细菌和真菌的污染的保存。载体可以是溶剂或分散介质，例如，水、乙醇、多元醇(如甘油、丙二醇、液态聚乙二醇)、植物油，及其适当的混合物。The present invention provides pharmaceutical compositions suitable for injection use, including sterile aqueous solutions or dispersion systems. Further, the above-mentioned pharmaceutical composition can be prepared in the form of a sterile powder that can be used for immediate preparation of sterile injection. In any case, the final injection form must be sterile, and for easy injection, it must be easy to flow. In addition, the pharmaceutical composition must be stable during preparation and storage. Therefore, preservation against contamination by microorganisms such as bacteria and fungi is preferred. The carrier can be a solvent or dispersion medium, for example, water, ethanol, polyol (such as glycerol, propylene glycol, liquid polyethylene glycol), vegetable oil, and suitable mixtures thereof.
本发明提供的药物组合物，可以是适于局部用药的形式，例如，气溶胶、乳剂、软膏、洗液、撒粉，或其他类似的剂型。进一步地，本发明提供的药物组合物可以采用适于经皮给药装置使用的形式。利用本发明式(I)所示化合物，或其药学上可接受的盐，通过常规的加工方法，可以制备这些制剂。作为一个例子，乳剂或软膏剂的制备是通过在上述化合物中加入亲水性材料和水(二者总量约为化合物的5wt％到50wt％)，制得具有预期一致性的乳剂或软膏。The pharmaceutical composition provided by the present invention may be in a form suitable for topical administration, for example, aerosol, emulsion, ointment, lotion, dusting, or other similar dosage forms. Further, the pharmaceutical composition provided by the present invention can be in a form suitable for use in a transdermal drug delivery device. These preparations can be prepared by using the compound represented by the formula (I) of the present invention, or a pharmaceutically acceptable salt thereof, through conventional processing methods. As an example, an emulsion or ointment is prepared by adding a hydrophilic material and water (the total amount of the two is about 5 wt% to 50 wt% of the compound) to prepare a cream or ointment with the desired consistency.
本发明提供的药物组合物，可以制成以固体为载体、适用于直肠给药的形式。混合物形成单位剂量的栓剂是最优选的剂型。适当的辅料包括本领域常用的可可脂和其他材料。栓剂可以方便地制备，首先药物组合物与软化或熔化的辅料混合，然后冷却和模具成型而制得。The pharmaceutical composition provided by the present invention can be made into a form suitable for rectal administration with a solid as a carrier. Suppositories in which the mixture forms a unit dose are the most preferred dosage form. Suitable excipients include cocoa butter and other materials commonly used in the art. Suppositories can be conveniently prepared. First, the pharmaceutical composition is mixed with softened or melted excipients, then cooled and molded.
除了上述提到的载体组分外，上述药学制剂还可以包括，适当的，一种或多种附加的辅料组分，如稀释剂、缓冲剂、调味剂、粘合剂、表面活性剂、增稠剂、润滑剂、防腐剂(包括抗氧化剂)等。进一步地，其他的辅药还可以包括调节药物与血液等渗压的促渗剂。包含有式(I)所示化合物，或其药学上可接受的盐的药物组合物，也可以制备成粉剂或浓缩液的形式。In addition to the above-mentioned carrier components, the above-mentioned pharmaceutical preparations may also include, as appropriate, one or more additional adjuvant components, such as diluents, buffers, flavoring agents, binders, surfactants, and additives. Thickeners, lubricants, preservatives (including antioxidants), etc. Further, other adjuvants may also include penetration enhancers that regulate the isotonic pressure of the drug and blood. The pharmaceutical composition containing the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, can also be prepared in the form of a powder or a concentrated solution.
具体实施方式detailed description
为使上述内容更清楚、明确，本发明将用以下实施例来进一步阐述本发明的技术方案。以下实施例仅用于说明本发明的具体实施方式，以使本领域的技术人员能够理解本发明，但不用于限制本发明的保护范围。本发明的具体实施方式中，未作特别说明的技术手段或方法等为本领域的常规技术手段或方法等。In order to make the above content clearer and clearer, the present invention will use the following embodiments to further illustrate the technical solution of the present invention. The following examples are only used to illustrate specific implementations of the present invention, so that those skilled in the art can understand the present invention, but are not used to limit the protection scope of the present invention. In the specific embodiments of the present invention, technical means or methods that are not specifically described are conventional technical means or methods in the art.
除非另有说明，本发明所有的一部分和百分比均按重量计算，所有温度均指摄氏度。Unless otherwise stated, all parts and percentages in the present invention are calculated by weight, and all temperatures refer to degrees Celsius.
实施例中使用了下列缩略语：The following abbreviations are used in the examples:
ACE-Cl：1-氯乙基氯甲酸酯；ACE-Cl: 1-chloroethyl chloroformate;
(BOC) 2O：二碳酸二叔丁酯； (BOC) 2 O: di-tert-butyl dicarbonate;
BOP：苯并三氮唑-1-基氧基三(二甲基氨基)磷鎓六氟磷酸盐；BOP: Benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate;
DBU：1,8-二氮杂二环十一碳-7-烯；DBU: 1,8-diazabicycloundec-7-ene;
DCE：1,2-二氯乙烷；DCE: 1,2-dichloroethane;
DCM：二氯甲烷；DCM: dichloromethane;
DIPEA或DIEA：N,N-二异丙基乙胺；DIPEA or DIEA: N,N-diisopropylethylamine;
DMAc：N,N-二甲基乙酰胺；DMAc: N,N-dimethylacetamide;
DMF：N,N-二甲基甲酰胺；DMF: N,N-dimethylformamide;
DMSO：二甲基亚砜；DMSO: dimethyl sulfoxide;
EDCI：1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐；EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride;
EtOAc或EA：乙酸乙酯；EtOAc or EA: ethyl acetate;
EtOH：乙醇；EtOH: ethanol;
EtONa：乙醇钠；EtONa: sodium ethoxide;
h、hr或hrs：小时；h, hr or hrs: hour;
Hex：正己烷；Hex: n-hexane;
HOBT：1-羟基苯并***；HOBT: 1-hydroxybenzotriazole;
LC-MS或LCMS：液相色谱-质谱联用；LC-MS or LCMS: liquid chromatography-mass spectrometry;
LDA：二异丙基氨基锂；LDA: lithium diisopropylamide;
MeCN：乙腈；MeCN: Acetonitrile;
MeOH：甲醇；MeOH: methanol;
MeONa：甲醇钠；MeONa: Sodium methoxide;
min或mins：分钟；min or mins: minutes;
MsCl：甲烷磺酰氯；MsCl: methanesulfonyl chloride;
MTBE：甲基叔丁基醚；MTBE: methyl tert-butyl ether;
MW：微波；MW: microwave;
NEt 3：三乙胺； NEt 3 : Triethylamine;
NBS：N-溴代琥珀酰亚胺；NBS: N-bromosuccinimide;
NMP：N-甲基-2-吡咯烷酮；NMP: N-methyl-2-pyrrolidone;
PdCl 2(dppf) 2：1,1’-双二苯基膦二茂铁二氯化钯； PdCl 2 (dppf) 2 :1,1'-bisdiphenylphosphinoferrocene palladium dichloride;
Pd 2(dba) 3：三(二亚苄基丙酮)二钯； Pd 2 (dba) 3 : Tris(dibenzylideneacetone)dipalladium;
Pd(OAc) 2：乙酸钯(II)； Pd(OAc) 2 : Palladium(II) acetate;
PE：石油醚；PE: petroleum ether;
PPA：多聚磷酸；PPA: polyphosphoric acid;
rt、r.t.或RT：室温；rt, r.t. or RT: room temperature;
TEA：三乙胺；TEA: triethylamine;
TFA：三氟乙酸；TFA: trifluoroacetic acid;
THF：四氢呋喃；THF: Tetrahydrofuran;
Ti(OEt) 4：钛酸四乙酯； Ti(OEt) 4 : Tetraethyl titanate;
TLC：薄层色谱；TLC: thin layer chromatography;
TMEDA：四甲基乙二胺；和TMEDA: Tetramethylethylenediamine; and
xantphos：4,5-双二苯基膦-9,9-二甲基氧杂蒽。xantphos: 4,5-bisdiphenylphosphine-9,9-dimethylxanthene.
中间体化合物M1的制备：Preparation of intermediate compound M1:
Figure PCTCN2020072774-appb-000013
Figure PCTCN2020072774-appb-000013
步骤1：化合物M1-3的制备Step 1: Preparation of compound M1-3
将15.00g化合物M1-1和7.08g化合物M1-2溶解于150mL二氧六环中，加入198mg Pd(OAc) 2、1.70g Xantphos和15.00g DIEA。氮气置换三次，氮气保护下反应升至85℃搅拌反应12hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(50mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得20.00g化合物M1-3。 15.00 g of compound M1-1 and 7.08 g of compound M1-2 were dissolved in 150 mL of dioxane, and 198 mg of Pd(OAc) 2 , 1.70 g of Xantphos and 15.00 g of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 85°C and stirred for 12 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (50 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 20.00 g of compound M1-3.
步骤2：化合物M1的制备Step 2: Preparation of compound M1
将20.00g化合物M1-3溶解于200mL THF中，-30℃下滴加EtONa(35mL，20％的EtOH溶液)，RT搅拌反应3hrs。TLC检测反应完全，减压浓缩，加入200mL的DCM搅拌30mins，反应液过滤，滤饼用DCM(50mL×2)洗涤，得固体15g化合物M1。20.00g of compound M1-3 was dissolved in 200mL THF, EtONa (35mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 3hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 200 mL of DCM and stirred for 30 mins, the reaction solution was filtered, and the filter cake was washed with DCM (50 mL×2) to obtain 15 g of compound M1 as a solid.
中间体化合物M2的制备：Preparation of intermediate compound M2:
Figure PCTCN2020072774-appb-000014
Figure PCTCN2020072774-appb-000014
步骤1：化合物M2-2的制备Step 1: Preparation of compound M2-2
将1.00g化合物M2-1溶解于10mL的DMSO中，加入MeONa的MeOH溶液(15mL，0.5M)，然后70℃反应1hr。TLC检测反应完全，将反应液倒入30mL水中，加EtOAc萃取(40mL×3)，合并有机相，用50mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得1.10g淡黄色油状物化合物M2-2。1.00 g of compound M2-1 was dissolved in 10 mL of DMSO, and MeONa in MeOH solution (15 mL, 0.5 M) was added, and then reacted at 70° C. for 1 hr. TLC detected the completion of the reaction. The reaction solution was poured into 30mL water, extracted with EtOAc (40mL×3), combined the organic phases, washed with 50mL saturated NaCl, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography. 1.10g of compound M2-2 as a pale yellow oil.
步骤2：化合物M2-3的制备Step 2: Preparation of compound M2-3
将化合物1.10g M2-2溶解于15mL二氧六环中，加入491mg化合物M1-2、46mg Xantphos、35mg Pd(OAc) 2和1.05g DIEA，混合物用氮气置换3次，加热至90℃反应5hrs。TLC检测反应完全，反应液冷却至室温，过滤，滤饼用EtOAc(5mL×3)洗涤，滤液减压浓缩，残余物经柱层析纯化得1.03g淡黄色固体M2-3。 Dissolve compound 1.10g M2-2 in 15mL dioxane, add 491mg compound M1-2, 46mg Xantphos, 35mg Pd(OAc) 2 and 1.05g DIEA, replace the mixture with nitrogen 3 times, heat to 90°C for 5hrs . TLC detected that the reaction was complete, the reaction solution was cooled to room temperature, filtered, the filter cake was washed with EtOAc (5 mL×3), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 1.03 g of light yellow solid M2-3.
步骤3：化合物M2的制备Step 3: Preparation of compound M2
将1.03g化合物M2-3溶解于10mL无水THF中，降温至-30℃，将EtONa的EtOH溶液(2mL，20％)缓慢滴加入上述溶液中，-30℃搅拌反应30mins，缓慢升至室温搅拌反应2hrs。TLC检测反应完全，将反应液减压浓缩，残余物加入20mL DCM打浆30mins， 过滤，滤饼用DCM(5mL×3)洗涤，取滤饼真空干燥得990mg棕色固体M2。Dissolve 1.03g of compound M2-3 in 10mL of anhydrous THF, lower the temperature to -30°C, slowly add EtONa's EtOH solution (2mL, 20%) to the above solution, stir and react at -30°C for 30mins, and slowly rise to room temperature Stir the reaction for 2hrs. TLC detected the completion of the reaction, the reaction solution was concentrated under reduced pressure, and the residue was added 20mL DCM to be slurried for 30mins, filtered, the filter cake was washed with DCM (5mL×3), and the filter cake was vacuum-dried to obtain 990mg of brown solid M2.
中间体化合物M3的制备：Preparation of intermediate compound M3:
Figure PCTCN2020072774-appb-000015
Figure PCTCN2020072774-appb-000015
步骤1：化合物M3-2的制备Step 1: Preparation of compound M3-2
将3.00g化合物M3-1和1.60g化合物M1-2溶解于30mL二氧六环中，加入243mg Pd 2(dba) 3、384g Xantphos和3.40g DIPEA，氮气置换三次，反应液升至110℃搅拌反应3hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(30mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得4.80g化合物M3-2。 Dissolve 3.00g of compound M3-1 and 1.60g of compound M1-2 in 30mL of dioxane, add 243mg of Pd 2 (dba) 3 , 384g of Xantphos and 3.40g of DIPEA, replace with nitrogen three times, raise the reaction solution to 110℃ and stir. Reaction 3hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (30 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 4.80 g of compound M3-2.
步骤2：化合物M3的制备Step 2: Preparation of compound M3
将4.80g化合物M3-2溶解于50mL THF中，-30℃下滴加EtONa(7.4mL，20％的EtOH溶液)，RT搅拌反应2hrs。TLC检测反应完全，减压浓缩，加入50mL DCM搅拌30mins，反应液过滤，滤饼用DCM(10mL×2)洗涤，抽干得3.60g化合物M3。4.80g of compound M3-2 was dissolved in 50mL THF, EtONa (7.4mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 2hrs. TLC detected the completion of the reaction, concentrated under reduced pressure, added 50mL DCM and stirred for 30mins, the reaction solution was filtered, the filter cake was washed with DCM (10mL×2), and drained to obtain 3.60g of compound M3.
中间体化合物M4的制备：Preparation of intermediate compound M4:
Figure PCTCN2020072774-appb-000016
Figure PCTCN2020072774-appb-000016
步骤1：化合物M4-3的制备Step 1: Preparation of compound M4-3
将300mg化合物M4-1和174mg化合物M4-2溶解于10mL二氧六环中，加入4mg Pd(OAc) 2、34mg Xantphos和300mg DIEA。氮气置换三次，氮气保护下反应升至85℃搅拌反应12hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得266mg化合物M4-3。 300 mg of compound M4-1 and 174 mg of compound M4-2 were dissolved in 10 mL of dioxane, and 4 mg of Pd(OAc) 2 , 34 mg of Xantphos and 300 mg of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 85°C and stirred for 12 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 266 mg of compound M4-3.
步骤2：化合物M4的制备Step 2: Preparation of compound M4
将266mg化合物M4-3溶解于5mL THF中，-30℃下滴加EtONa(0.47mL，20％的EtOH溶液)，RT搅拌反应3hrs。TLC检测反应完全，减压浓缩，加入15mL的DCM搅拌30mins，反应液过滤，滤饼用DCM(20mL×2)洗涤，得固体182mg化合物M4。266 mg of compound M4-3 was dissolved in 5 mL of THF, EtONa (0.47 mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 3 hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 15 mL of DCM and stirred for 30 mins, the reaction solution was filtered, and the filter cake was washed with DCM (20 mL×2) to obtain 182 mg of compound M4 as a solid.
中间体化合物M5的制备：Preparation of intermediate compound M5:
Figure PCTCN2020072774-appb-000017
Figure PCTCN2020072774-appb-000017
步骤1：化合物M5-3的制备Step 1: Preparation of compound M5-3
氮气保护下，将10.60g化合物M5-2溶解于26mL乙醇中，反应液降温0℃，滴加25.00g的M5-1，滴完后撤除冰浴RT反应2hrs，然后加热90℃反应过夜。TLC检测反应完全，反应液减压浓缩，残余物经柱层析纯化得白色固体4.94g，即化合物M5-3。Under the protection of nitrogen, 10.60g of compound M5-2 was dissolved in 26mL of ethanol, the reaction solution was cooled to 0°C, and 25.00g of M5-1 was added dropwise. After dropping, the ice bath RT was removed for 2hrs, and then heated at 90°C for overnight reaction. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 4.94 g of a white solid, namely compound M5-3.
步骤2：化合物M5的制备Step 2: Preparation of compound M5
将化合物M5-3溶于100mL DMF中，降温至0℃，一次性加入4.98g NBS，然后RT反应1hr。TLC检测反应完全，加300mL水淬灭反应，EtOAc(300mL×2)萃取，合并有机相，饱和NaCl(50mL×4)洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得5.58g白色固体，即化合物M5。The compound M5-3 was dissolved in 100 mL DMF, the temperature was lowered to 0°C, 4.98 g NBS was added all at once, and the reaction was carried out at RT for 1 hr. The reaction was completed by TLC detection, the reaction was quenched by adding 300 mL of water, extracted with EtOAc (300 mL×2), combined the organic phases, washed with saturated NaCl (50 mL×4), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography 5.58 g of white solid, compound M5 was obtained.
中间体化合物M6的制备：Preparation of intermediate compound M6:
Figure PCTCN2020072774-appb-000018
Figure PCTCN2020072774-appb-000018
步骤1：化合物M6-3的制备Step 1: Preparation of compound M6-3
氮气保护下，将25.00g化合物M6-1溶解于200mL的DMF中，降温至0℃，分批加入22.70g NaH，0℃保温1hr，然后将54.96g化合物M6-2缓慢滴加到反应液中，滴完后0℃下反应1hr，升温至60℃继续反应1hr。反应液降温至0℃，用500mL冰水淬灭反应，EtOAc(500mL×3)萃取，合并有机相，有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得29.00g棕色油状物M6-3。Under the protection of nitrogen, dissolve 25.00g of compound M6-1 in 200mL of DMF, reduce the temperature to 0℃, add 22.70g NaH in batches, keep at 0℃ for 1hr, and then slowly add 54.96g compound M6-2 dropwise to the reaction solution After dropping, react at 0°C for 1 hr, and then heat to 60°C to continue the reaction for 1 hr. The reaction solution was cooled to 0°C, quenched with 500 mL ice water, extracted with EtOAc (500 mL×3), combined the organic phases, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was passed through the column Analyze and purify to obtain 29.00g brown oil M6-3.
步骤2：化合物M6-5的制备Step 2: Preparation of compound M6-5
将29.00g化合物M6-3溶解于50mL的Ti(OEt) 4中，加入34.99g化合物M6-4，然后加热至90℃反应12hrs。TCL检测反应完全，将反应液倒入500mL的冰水中，加入300mL  EtOAc搅拌1hr，用EtOAc(300mL×3)萃取，合并有机相，有机相用饱和食盐水(100mL×4)洗涤，无水硫酸钠干燥，减压浓缩得39.00g棕色油状物化合物M6-5粗品。 29.00g of compound M6-3 was dissolved in 50 mL of Ti(OEt) 4 , 34.99g of compound M6-4 was added, and then heated to 90°C for 12 hrs. TCL detected that the reaction was complete. Pour the reaction solution into 500 mL ice water, add 300 mL EtOAc and stir for 1 hr, extract with EtOAc (300 mL×3), combine the organic phases, and wash the organic phase with saturated brine (100 mL×4), anhydrous sulfuric acid It was dried with sodium and concentrated under reduced pressure to obtain 39.00 g of crude compound M6-5 as a brown oil.
步骤3：化合物M6-6的制备Step 3: Preparation of compound M6-6
氮气保护下，将48.00g化合物M6-5溶解于500mL无水THF中，降温至-20℃，缓慢加入6.73g NaHB 4，然后自然升温至RT搅拌2hrs。反应完毕，反应液降温至0℃，用300mL水淬灭，EtOAc(300mL×3)萃取，合并有机相，有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得25.40g棕色油状物的化合物M6-6。 Under the protection of nitrogen, 48.00g of compound M6-5 was dissolved in 500mL of anhydrous THF, the temperature was lowered to -20°C, 6.73g of NaHB 4 was slowly added, and then the temperature was raised to RT and stirred for 2hrs. After the reaction, the reaction solution was cooled to 0°C, quenched with 300 mL of water, extracted with EtOAc (300 mL×3), combined the organic phases, washed with saturated brine, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was passed through a column Chromatographic purification yielded 25.40 g of compound M6-6 as a brown oil.
步骤4：化合物M6的制备Step 4: Preparation of compound M6
将10.00g化合物M6-6溶解于100mL DCM溶液中，滴加28.04g TFA溶液，然后RT下反应1hr。反应液降温至0℃，用100mL饱和NaHCO 3水溶液淬灭，EtOAc:THF＝3:1(100mL×3)萃取，合并有机相，有机相用饱和食盐水洗涤，无水硫酸钠干燥，减压浓缩得7.64g棕色固体即化合物M6粗品，直接用于下一步反应。 10.00 g of compound M6-6 was dissolved in 100 mL of DCM solution, 28.04 g of TFA solution was added dropwise, and then reacted at RT for 1 hr. The reaction solution was cooled to 0°C, quenched with 100 mL saturated aqueous NaHCO 3 , extracted with EtOAc:THF=3:1 (100 mL×3), combined the organic phases, washed with saturated brine, dried over anhydrous sodium sulfate, and reduced pressure It was concentrated to obtain 7.64 g of a brown solid, the crude compound M6, which was directly used in the next reaction.
1H NMR(500MHz,DMSO-d 6)δ7.26-7.21(m,4H),5.80(d,J＝10.5Hz,1H),4.43(d,J＝10.5Hz,1H),3.17-3.15(m,2H),3.08(d,J＝15.5Hz,1H),2.98-2.88(m,2H),2.69(d,J＝15.5Hz,1H),2.04-1.99(m,1H),1.80-1.75(m,1H),1.62-1.59(m,1H),1.35(m,1H),1.22(s,9H)。 1 H NMR(500MHz,DMSO-d 6 )δ7.26-7.21(m,4H), 5.80(d,J=10.5Hz,1H), 4.43(d,J=10.5Hz,1H), 3.17-3.15( m, 2H), 3.08 (d, J = 15.5 Hz, 1H), 2.98-2.88 (m, 2H), 2.69 (d, J = 15.5 Hz, 1H), 2.04-1.99 (m, 1H), 1.80-1.75 (m, 1H), 1.62-1.59 (m, 1H), 1.35 (m, 1H), 1.22 (s, 9H).
中间体化合物M7的制备：Preparation of intermediate compound M7:
Figure PCTCN2020072774-appb-000019
Figure PCTCN2020072774-appb-000019
步骤1：化合物M7-3的制备Step 1: Preparation of compound M7-3
将10.00g化合物M7-1和19.50g化合物M7-2溶解于100mL的MeCN中，加入26.20g K 2CO 3。反应升至90℃搅拌反应3hrs。TLC检测反应完全，将反应液过滤，滤饼用EtOAc(50mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得5.90g化合物M7-3。 10.00 g of compound M7-1 and 19.50 g of compound M7-2 were dissolved in 100 mL of MeCN, and 26.20 g of K 2 CO 3 was added . The reaction was raised to 90°C and stirred for 3hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with EtOAc (50 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 5.90 g of compound M7-3.
步骤2：化合物M7-4的制备Step 2: Preparation of compound M7-4
将1.50g化合物M7-3溶解于15mL甲苯中，滴加1.1mL PBr 3，反应液升至105℃搅拌反应12hrs。TLC检测反应完全，减压浓缩，加入15mL水，用NaOH溶液调至pH＝9，用 EtOAc(30mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩得1.50g化合物M7-4。 1.50 g of compound M7-3 was dissolved in 15 mL of toluene, 1.1 mL of PBr 3 was added dropwise, the reaction solution was raised to 105° C. and stirred for 12 hrs. TLC detects the completion of the reaction, concentrated under reduced pressure, added 15 mL of water, adjusted to pH=9 with NaOH solution, extracted with EtOAc (30 mL×3), combined the organic phases, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 1.50 g of compound M7- 4.
步骤3：化合物M7-6的制备Step 3: Preparation of compound M7-6
将450mg化合物M7-5溶解于6mL DMF中，0℃下分批加入271mg NaH，氮气保护下，60℃搅拌反应1hr后加入1.20g化合物M4-4，60℃搅拌反应1hr。TLC检测反应完全，加入30mL水淬灭反应，用EtOAc(25mL×2)和水(30mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得160mg化合物M7-6。Dissolve 450 mg of compound M7-5 in 6 mL DMF, add 271 mg of NaH in batches at 0°C, stir and react at 60°C for 1 hr under nitrogen protection, then add 1.20 g of compound M4-4, and stir at 60°C for 1 hr. TLC detected the completion of the reaction. The reaction was quenched by adding 30 mL of water, and extracted with EtOAc (25 mL×2) and water (30 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography. 160 mg of compound M7-6.
步骤4：化合物M7-7的制备Step 4: Preparation of compound M7-7
在0℃下将160mg化合物M7-6溶解于2mL DCE中，滴加入155mg ACE-Cl，RT搅拌反应2hrs。TLC检测反应完全，减压浓缩，加入4mL MeOH，反应升至80℃搅拌反应3hrs。TLC检测反应完全，减压浓缩，加入4mL DCM，242mg(Boc) 2O和239mg DIEA，RT搅拌反应12hrs。TLC检测反应完全，减压浓缩，残余物经柱层析纯化得25mg化合物M7-7。 160 mg of compound M7-6 was dissolved in 2 mL of DCE at 0°C, 155 mg of ACE-Cl was added dropwise, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, 4mL MeOH was added, and the reaction was raised to 80°C and stirred for 3hrs. The reaction was completed by TLC detection, concentrated under reduced pressure, 4 mL DCM, 242 mg (Boc) 2 O and 239 mg DIEA were added, and the reaction was stirred at RT for 12 hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 25 mg of compound M7-7.
步骤5：化合物M7的制备Step 5: Preparation of compound M7
由化合物M7-7制备化合物M7的步骤类似于由化合物M6-3到化合物M6的步骤。The procedure for preparing compound M7 from compound M7-7 is similar to the procedure from compound M6-3 to compound M6.
中间体化合物M8的制备：Preparation of intermediate compound M8:
Figure PCTCN2020072774-appb-000020
Figure PCTCN2020072774-appb-000020
步骤1：化合物M8-3的制备Step 1: Preparation of compound M8-3
将4.00g化合物M8-1溶解于50mL无水THF中，氮气置换三次，降温至-78℃，缓慢滴加入LDA的THF溶液(11.70mL，2.0M)，然后-78℃反应1hr，将化合物M8-2的THF(10mL)溶液缓慢滴加入上述反应液中，-78℃反应30mins，缓慢升至室温反应2hrs。TLC检测反应完全，用30mL饱和NH 4Cl溶液淬灭反应，加入50mL水，加EtOAc萃取(60mL×3)，合并有机相，用50mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得4.30g无色油状物M8-3。 Dissolve 4.00g of compound M8-1 in 50mL of anhydrous THF, replace with nitrogen three times, cool to -78°C, slowly add LDA solution in THF (11.70mL, 2.0M) dropwise, and then react at -78°C for 1 hr to convert compound M8 -2 in THF (10 mL) was slowly added dropwise to the above reaction solution, reacted at -78°C for 30 mins, and slowly raised to room temperature for 2 hrs. TLC detected the completion of the reaction. The reaction was quenched with 30 mL saturated NH 4 Cl solution, 50 mL water was added, and EtOAc was added for extraction (60 mL×3). The organic phases were combined, washed with 50 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was purified by column chromatography to obtain 4.30 g of colorless oil M8-3.
步骤2：化合物M8-4的制备Step 2: Preparation of compound M8-4
将4.30g化合物M8-3溶解于THF/MeOH(40mL/40mL)中，加入NaOH水溶液(20mL， 2.4N)，加热至80℃反应18hrs。TLC检测反应完全，反应液冷却至室温，减压浓缩蒸除有机溶剂，残余物用浓盐酸调至pH为3-4，过滤，滤饼用水(10mL×3)洗涤，将滤饼进行真空干燥得白色固体3.40g化合物M8-4。4.30g of compound M8-3 was dissolved in THF/MeOH (40mL/40mL), NaOH aqueous solution (20mL, 2.4N) was added, and the reaction was heated to 80°C for 18hrs. TLC detected the completion of the reaction, the reaction solution was cooled to room temperature, concentrated under reduced pressure to remove the organic solvent, the residue was adjusted to pH 3-4 with concentrated hydrochloric acid, filtered, the filter cake was washed with water (10 mL×3), and the filter cake was vacuum dried 3.40 g of compound M8-4 was obtained as a white solid.
步骤3：化合物M8-5的制备Step 3: Preparation of compound M8-5
将3.40g化合物M8-4溶解于40mL PPA中，升温至120℃反应2hrs。TLC检测反应完全，将反应液缓慢滴加入200mL碎冰中，用2.4N NaOH水溶液调至pH为9-10，加入4.40g(B OC) 2O，RT搅拌反应18hrs。TLC检测反应完全，加EtOAc萃取(100mL×3)，合并有机相，用100mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得2.40g白色固体M8-5。 3.40g of compound M8-4 was dissolved in 40mL PPA, and the temperature was raised to 120°C to react for 2hrs. TLC detected that the reaction was complete. The reaction solution was slowly added dropwise to 200 mL crushed ice, adjusted to pH 9-10 with 2.4N NaOH aqueous solution, 4.40 g (B OC ) 2 O was added, and the reaction was stirred at RT for 18 hrs. TLC detected that the reaction was complete, extracted with EtOAc (100 mL×3), combined the organic phases, washed with 100 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 2.40 g of white solid M8-5.
步骤4：化合物M8的制备Step 4: Preparation of compound M8
由化合物M8-5制备化合物M8的步骤类似于由化合物M6-3到化合物M6的步骤。The procedure for preparing compound M8 from compound M8-5 is similar to the procedure from compound M6-3 to compound M6.
中间体M9-a和M9-b的制备：Preparation of intermediates M9-a and M9-b:
Figure PCTCN2020072774-appb-000021
Figure PCTCN2020072774-appb-000021
步骤1：化合物M9-2的制备Step 1: Preparation of compound M9-2
将10.00g化合物M9-1溶于100mL MeOH中，加入2.0mL浓硫酸，加热70℃反应3hrs。反应完毕后，旋干溶剂，加20mL水，用饱和Na 2CO 3水溶液调至pH＝9，EtOAc(100mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得9.23g白色固体，即化合物M9-2。 10.00 g of compound M9-1 was dissolved in 100 mL of MeOH, 2.0 mL of concentrated sulfuric acid was added, and the reaction was heated at 70° C. for 3 hrs. After the reaction is complete, spin off the solvent, add 20 mL of water, adjust to pH=9 with saturated aqueous Na 2 CO 3 , extract with EtOAc (100 mL×3), combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. Purification by column chromatography yielded 9.23 g of white solid, compound M9-2.
步骤2：化合物M9-3的制备Step 2: Preparation of compound M9-3
将9.23g化合物M9-2溶于150mL MeOH中，冰浴冷至0℃，分批加入6.97g NaBH 4，自然升温至RT反应5hrs。反应完毕后，加入20mL饱和NH 4Cl溶液，旋干溶剂，加入EtOAc(100mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得6.40g无色液体，即化合物M9-3。 Dissolve 9.23 g of compound M9-2 in 150 mL of MeOH, cool to 0° C. in an ice bath, add 6.97 g of NaBH 4 in batches, and naturally increase the temperature to RT for 5 hrs. After the completion of the reaction, add 20 mL of saturated NH 4 Cl solution, spin dry the solvent, add EtOAc (100 mL×3) for extraction, combine the organic phases, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by column chromatography to obtain 6.40 g Colored liquid, compound M9-3.
步骤3：化合物M9-4的制备Step 3: Preparation of compound M9-4
氮气保护下，将3.00g化合物M9-3溶于50mL二氯甲烷中，反应液降温至-15℃，加入2.81mL NEt 3，然后滴加1.04mL的MsCl溶液，滴加完毕后，升温至0℃反应1hr。反应完毕后，加入水分层，有机相用20mL饱和食盐水洗涤，有机相无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得4.05g白色固体的化合物M9-4。 Under nitrogen protection, dissolve 3.00g of compound M9-3 in 50mL of dichloromethane, cool the reaction solution to -15°C, add 2.81mL of NEt 3 , and then add 1.04mL of MsCl solution dropwise. After the addition is complete, the temperature is raised to 0 React at ℃ for 1hr. After the reaction was completed, a water layer was added, the organic phase was washed with 20 mL of saturated brine, the organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 4.05 g of white solid compound M9-4.
步骤4：化合物M9-6的制备Step 4: Preparation of compound M9-6
氮气保护下，将3.39g化合物M9-5溶于20mL无水THF中，降温至-50℃，滴加1.71g LDA溶液，滴完后-50℃保温反应1hr后。滴加3.00g化合物M9-4的无水THF(10mL)溶液，滴毕，升温至RT反应1hr。反应完毕后，加入50mL食盐水，EtOAc(50mL×3)萃取，合并有机相，有机相用水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得4.61g白色固体的化合物M9-6。Under the protection of nitrogen, dissolve 3.39 g of compound M9-5 in 20 mL of anhydrous THF, lower the temperature to -50°C, and add 1.71 g of LDA solution dropwise. After dripping, keep the temperature at -50°C and react for 1 hour. A solution of 3.00 g of compound M9-4 in anhydrous THF (10 mL) was added dropwise, after the dropping, the temperature was raised to RT and reacted for 1 hr. After the reaction was completed, 50 mL brine was added, extracted with EtOAc (50 mL×3), the organic phases were combined, the organic phase was dried with sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 4.61 g of white solid compound M9-6.
步骤5：化合物M9-7a和M9-7b的制备Step 5: Preparation of compounds M9-7a and M9-7b
将4.61g化合物M9-6溶于8mL水和40mL MeOH中，加入2.07g NaOH。升温至65℃搅拌过夜。反应完毕后，加入30mL水，旋干溶剂甲醇，浓缩物用2N盐酸调至pH＝6，EtOAc(50mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得3.98g白色固体，即化合物M9-7a和M9-7b。Dissolve 4.61g of compound M9-6 in 8mL of water and 40mL of MeOH, and add 2.07g of NaOH. The temperature was raised to 65°C and stirred overnight. After the reaction is complete, add 30 mL of water, spin dry the solvent methanol, adjust the concentrate to pH=6 with 2N hydrochloric acid, extract with EtOAc (50 mL×3), combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and pass the residue through a column Chromatographic purification yielded 3.98 g of white solids, namely compounds M9-7a and M9-7b.
步骤6：化合物M9-8a和M9-8b的制备Step 6: Preparation of compounds M9-8a and M9-8b
氮气保护下，将3.98g混合物M9-7a和M9-7b溶于20mL无水THF中，反应液降温至-15℃，分批加入NaH(60％，0.42g)，然后-15℃保温反应1hr，然后降温至-60℃，滴加正丁基锂(1.6M，7.8mL)，保温反应1hr。反应完毕，加入50mL水，用EtOAc(50mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得2.36g淡黄色固体，即混合物M9-8a和M9-8b。Under the protection of nitrogen, dissolve 3.98g of the mixture of M9-7a and M9-7b in 20mL of anhydrous THF, cool the reaction solution to -15℃, add NaH (60%, 0.42g) in batches, and then keep at -15℃ for 1hr. Then, the temperature was lowered to -60°C, n-butyllithium (1.6M, 7.8mL) was added dropwise, and the reaction was kept warm for 1hr. After the reaction was completed, 50 mL of water was added, extracted with EtOAc (50 mL×3), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 2.36 g of pale yellow solid, which is a mixture of M9-8a and M9-8b.
步骤7：化合物M9-a和M9-b的制备Step 7: Preparation of compounds M9-a and M9-b
由M9-8a和M9-8b制备M9-a和M9-b的步骤类似于由化合物M6-3到化合物M6的步骤。The procedure for preparing M9-a and M9-b from M9-8a and M9-8b is similar to the procedure from compound M6-3 to compound M6.
中间体化合物M10的制备：Preparation of intermediate compound M10:
Figure PCTCN2020072774-appb-000022
Figure PCTCN2020072774-appb-000022
步骤1：化合物M10-3的制备Step 1: Preparation of compound M10-3
氮气保护下，将2.83g化合物M10-2溶解于50mL的无水THF中，降温至-78℃，滴加LDA(2M，6mL)的THF/Hex溶液。在-78℃保温1hr，然后将1.69g化合物M10-1的THF(3mL)溶液缓慢滴加到反应液中，滴完后-78℃下反应1hr。反应液用50mL饱和食盐水淬灭反应，EtOAc(30mL×2)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得1.44g淡黄色油状物化合物M10-3。Under the protection of nitrogen, 2.83 g of compound M10-2 was dissolved in 50 mL of anhydrous THF, the temperature was lowered to -78° C., and a THF/Hex solution of LDA (2M, 6 mL) was added dropwise. Incubate at -78°C for 1 hr, then slowly drop 1.69 g of compound M10-1 in THF (3 mL) into the reaction solution, and react at -78°C for 1 hr. The reaction solution was quenched with 50 mL saturated brine, extracted with EtOAc (30 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 1.44 g of a pale yellow oil compound M10- 3.
步骤2：化合物M10-4的制备Step 2: Preparation of compound M10-4
氮气保护下，将900mg化合物M10-3溶解于50mL无水THF中，降温至-78℃。滴加LDA(2M，3mL)的THF/Hex溶液。在-78℃保温反应1hr。反应液用50mL饱和食盐水淬灭反应，EtOAc(30mL×2)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得300mg淡黄色固体化合物M10-4。Under the protection of nitrogen, 900 mg of compound M10-3 was dissolved in 50 mL of anhydrous THF, and the temperature was lowered to -78°C. A THF/Hex solution of LDA (2M, 3 mL) was added dropwise. The reaction was incubated at -78°C for 1 hr. The reaction solution was quenched with 50 mL saturated brine, extracted with EtOAc (30 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 300 mg of light yellow solid compound M10-4.
步骤3：化合物M10的制备Step 3: Preparation of compound M10
由化合物M10-4制备化合物M10的步骤类似于由化合物M6-3到化合物M6的步骤。The procedure for preparing compound M10 from compound M10-4 is similar to the procedure from compound M6-3 to compound M6.
经由不同的反应起始原料和合适的试剂，例如合成M12的起始原料为6-甲氧基-1-茚酮，采用与前述中间体6-10类似的方法制备表1中的中间体化合物M11-M15。Through different reaction starting materials and suitable reagents, for example, the starting material for the synthesis of M12 is 6-methoxy-1-indanone, the intermediate compounds in Table 1 are prepared by a method similar to the aforementioned intermediate 6-10 M11-M15.
表1Table 1
Figure PCTCN2020072774-appb-000023
Figure PCTCN2020072774-appb-000023
实施例2：化合物(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫代)-5-甲基吡嗪-2-基)甲醇的制备Example 2: Compound (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3- Preparation of dichlorophenyl)thio)-5-methylpyrazin-2-yl)methanol
Figure PCTCN2020072774-appb-000024
Figure PCTCN2020072774-appb-000024
步骤1：化合物2-2的制备Step 1: Preparation of compound 2-2
将500mg化合物2-1和729mg化合物M5溶解于15mL二氧六环中，加入127mg Pd 2(dba) 3、161mg Xantphos和1g DIEA。氮气置换三次，氮气保护下在80℃搅拌12hrs。TLC检测反应完全，将反应液过滤，滤饼用EtOAc(15mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得350mg化合物2-2。 500 mg of compound 2-1 and 729 mg of compound M5 were dissolved in 15 mL of dioxane, and 127 mg of Pd 2 (dba) 3 , 161 mg of Xantphos and 1 g of DIEA were added. Replace with nitrogen three times and stir at 80°C for 12 hrs under nitrogen protection. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with EtOAc (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 350 mg of compound 2-2.
步骤2：化合物2-3的制备Step 2: Preparation of compound 2-3
将228mg化合物2-2溶解于3mL POCl 3中，加入38mg化合物N,N-二甲基苯酰胺，反应升至110℃搅拌1.5hrs。TLC检测反应完全，减压浓缩，在冰浴下加NaHCO 3水溶液调至pH为7-8，EtOAc/THF＝1/1(30mL×2)萃取，合并有机相，无水硫酸钠干燥，减压浓缩得195mg化合物2-3。 228 mg of compound 2-2 was dissolved in 3 mL of POCl 3 , and 38 mg of compound N,N-dimethylbenzamide was added, and the reaction was raised to 110° C. and stirred for 1.5 hrs. TLC detects the completion of the reaction, concentrates under reduced pressure, adds NaHCO 3 aqueous solution to adjust the pH to 7-8 in an ice bath, and extracts with EtOAc/THF=1/1 (30mL×2). Combine the organic phases, dry with anhydrous sodium sulfate, and reduce Press and concentrate to obtain 195 mg of compound 2-3.
步骤3：化合物2-4的制备Step 3: Preparation of compound 2-4
将195mg化合物2-3和236mg化合物M6溶解于3mL NMP中，加入268mg DIEA。反应升至80℃搅拌3hrs。TLC检测反应完全，EtOAc/THF＝1/1(25mL×2)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得165mg化合物2-4。Dissolve 195mg of compound 2-3 and 236mg of compound M6 in 3mL NMP, and add 268mg DIEA. The reaction was raised to 80°C and stirred for 3hrs. TLC detected that the reaction was complete, extracted with EtOAc/THF=1/1 (25 mL×2), combined the organic phases, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 165 mg of compound 2-4.
步骤4：化合物2-5的制备Step 4: Preparation of compound 2-5
在0℃下将165mg化合物2-4溶解于2.5mL THF中，分批加入24mg LiAlH 4，搅拌10mins。TLC检测反应完全，0℃下依次加入50μL水、NaOH(15％，50μL)、150μL水和1g无水硫酸钠，搅拌10mins。过滤，滤饼用THF/DCM＝1/1(10mL×2)洗涤，滤液减 压浓缩，残余物经柱层析纯化得94mg化合物2-5。 165 mg of compound 2-4 was dissolved in 2.5 mL THF at 0° C., 24 mg LiAlH 4 was added in batches, and stirred for 10 mins. TLC detected that the reaction was complete, and 50 μL of water, NaOH (15%, 50 μL), 150 μL of water and 1 g of anhydrous sodium sulfate were sequentially added at 0° C., and stirred for 10 mins. After filtration, the filter cake was washed with THF/DCM=1/1 (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 94 mg of compound 2-5.
步骤5：化合物2的制备Step 5: Preparation of compound 2
将94mg化合物2-5溶解于1.5mL二氧六烷中，加入0.4mL 2N HCl的甲醇溶液，RT反应1hr。TLC检测反应完全，减压浓缩，加NaHCO 3水溶液调至pH为7-8，DCM/MeOH＝10/1(15mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得47.9mg化合物2。 94 mg of compound 2-5 was dissolved in 1.5 mL of dioxane, 0.4 mL of 2N HCl in methanol was added, and reacted at RT for 1 hr. TLC detects the completion of the reaction, concentrates under reduced pressure, adds aqueous NaHCO 3 to adjust the pH to 7-8, extracts with DCM/MeOH = 10/1 (15 mL×3), combines the organic phases, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The product was purified by column chromatography to obtain 47.9 mg of compound 2.
[M+H +]＝501.13。 [M+H + ]=501.13.
1H NMR(500MHz,DMSO-d 6):δ7.48(d,J＝8.5Hz,1H),7.32-7.30(m,1H),7.25(t,J＝8.5Hz,1H),7.18-7.13(m,3H),6.77(d,J＝8.5Hz,1H),5.35(t,J＝5.5Hz,1H),4.46(d,J＝5.5Hz,1H),3.90-3.83(m,3H),3.18-3.09(m,2H),3.04(d,J＝16.0Hz,1H),2.62(d,J＝16.0Hz,1H),2.41(s,3H),1.92-1.86(m,1H),1.80-1.75(m,1H),1.55-1.53(m,1H),1.16-1.13(m,1H)。 1 H NMR (500MHz, DMSO-d 6 ): δ7.48 (d, J=8.5Hz, 1H), 7.32-7.30 (m, 1H), 7.25 (t, J=8.5Hz, 1H), 7.18-7.13 (m, 3H), 6.77 (d, J = 8.5 Hz, 1H), 5.35 (t, J = 5.5 Hz, 1H), 4.46 (d, J = 5.5 Hz, 1H), 3.90-3.83 (m, 3H) ,3.18-3.09(m,2H),3.04(d,J=16.0Hz,1H), 2.62(d,J=16.0Hz,1H),2.41(s,3H),1.92-1.86(m,1H), 1.80-1.75 (m, 1H), 1.55-1.53 (m, 1H), 1.16-1.13 (m, 1H).
实施例3：化合物(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲腈的制备Example 3: Compound (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazine-2-carbonitrile preparation
Figure PCTCN2020072774-appb-000025
Figure PCTCN2020072774-appb-000025
步骤1：化合物3-2的制备Step 1: Preparation of compound 3-2
RT下，将174mg化合物3-1和306mg化合物M6溶于3mL无水THF中，加入0.28mL NEt 3。氮气置换三次后RT反应1hr。反应完毕，加入10mL水和10mL EtOAc萃取三次，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得142mg黄色油状物的化合物3-2。 At RT, 174 mg of compound 3-1 and 306 mg of compound M6 were dissolved in 3 mL of anhydrous THF, and 0.28 mL of NEt 3 was added. After nitrogen replacement for three times, RT reaction for 1 hr. After the reaction was completed, 10 mL of water and 10 mL of EtOAc were added for extraction three times, the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 142 mg of yellow oily compound 3-2.
步骤2：化合物3-3的制备Step 2: Preparation of compound 3-3
将142mg化合物3-2、62.3mg化合物M1和55mg KI溶于二氧六环中，微波120℃反应1hr。LC-MS检测反应完全，加入10mL水和10mL的EtOAc萃取三次，合并有机层，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得121mg黄色固体的化合物3-3。Dissolve 142 mg of compound 3-2, 62.3 mg of compound M1 and 55 mg of KI in dioxane and react in microwave at 120°C for 1 hr. LC-MS detected that the reaction was complete, 10 mL of water and 10 mL of EtOAc were added for extraction three times, the organic layers were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 121 mg of yellow solid compound 3-3.
步骤3：化合物3的制备Step 3: Preparation of compound 3
将60mg化合物3-3溶于2.0mL的1,4-二氧六环中，RT条件下，滴加0.3mL的2M盐酸的甲醇溶液，RT条件下反应30mins。反应完全后，旋干溶剂，加入1mL水，用饱和NaHCO 3水溶液调至pH为8，析出固体后加10mL水，用EtOAc(10mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得30.2mg黄色固体的化合物3。 60 mg of compound 3-3 was dissolved in 2.0 mL of 1,4-dioxane, 0.3 mL of 2M hydrochloric acid in methanol was added dropwise under RT conditions, and reacted for 30 mins under RT conditions. After the reaction is complete, spin off the solvent, add 1 mL of water, adjust the pH to 8 with saturated NaHCO 3 aqueous solution, add 10 mL of water after the solid is precipitated, and extract with EtOAc (10 mL×3). Combine the organic phases and dry with anhydrous sodium sulfate. After pressure concentration, the residue was purified by column chromatography to obtain 30.2 mg of compound 3 as a yellow solid.
[M+H +]＝464.19。 [M+H + ]=464.19.
1H NMR(500MHz,CDCl3):δ8.34(s,1H),7.79(d,J＝5.5Hz,1H),7.35-7.33(m,1H),7.25(m,3H),6.17(d,J＝5.5Hz,1H),4.56-4.50(m,2H),4.03(s,1H),3.51-3.42(m,2H),3.12(d,J＝15.5Hz,1H),2.77(d,J＝15.5Hz,1H),2.01-1.94(m,1H),1.91-1.83(m,1H),1.47-1.44(m,1H),1.26-1.24(m,1H)。 1 H NMR (500MHz, CDCl3): δ8.34 (s, 1H), 7.79 (d, J = 5.5Hz, 1H), 7.35-7.33 (m, 1H), 7.25 (m, 3H), 6.17 (d, J = 5.5Hz, 1H), 4.56-4.50 (m, 2H), 4.03 (s, 1H), 3.51-3.42 (m, 2H), 3.12 (d, J = 15.5Hz, 1H), 2.77 (d, J = 15.5 Hz, 1H), 2.01-1.94 (m, 1H), 1.91-1.83 (m, 1H), 1.47-1.44 (m, 1H), 1.26-1.24 (m, 1H).
实施例4：化合物(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫基)吡嗪-2-甲酰胺的制备Example 4: Compound (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-bis Preparation of chlorophenyl)thio)pyrazine-2-carboxamide
Figure PCTCN2020072774-appb-000026
Figure PCTCN2020072774-appb-000026
步骤1：化合物4-2的制备Step 1: Preparation of compound 4-2
向200mg化合物4-1和104.86mg化合物2-1的DMA(3mL)溶液中加入14.96mg KI和186.77mg K 2CO 3，120℃下搅拌2hrs。反应液用10mL水稀释，EtOAc(10mL×3)萃取，合并有机相，有机相用饱和食盐水洗涤，无水硫酸钠干燥后浓缩，粗品经Pre-TLC(PE:EtOAc＝3:1)分离纯化，得到233mg化合物4-2为黄色固体，产率为88.18％。 14.96 mg of KI and 186.77 mg of K 2 CO 3 were added to the DMA (3 mL) solution of 200 mg of compound 4-1 and 104.86 mg of compound 2-1, and stirred at 120°C for 2 hrs. The reaction solution was diluted with 10 mL of water, extracted with EtOAc (10 mL×3), and the organic phases were combined. The organic phase was washed with saturated brine, dried over anhydrous sodium sulfate and concentrated. The crude product was separated by Pre-TLC (PE:EtOAc=3:1) After purification, 233 mg of compound 4-2 was obtained as a yellow solid with a yield of 88.18%.
[M+H +]:586.23。 [M+H + ]:586.23.
步骤2：化合物4-3的制备Step 2: Preparation of compound 4-3
向200mg化合物4-2的二氧六环(1.56mL)和水(1.56mL)的混合溶液中滴加NaOH(2.5M,132.97μL)，100℃下搅拌12hrs。反应液用1N HCl调节pH为6-7，EtOAc(10mL×3)萃取，合并有机相，有机相用饱和食盐水洗涤，无水硫酸钠干燥后浓缩，粗品经Pre-TLC(DCM:MeOH＝20:1)分离纯化，得到93mg化合物4-3为黄色固体，产率为69.41％。NaOH (2.5M, 132.97 μL) was added dropwise to a mixed solution of 200 mg of compound 4-2 in dioxane (1.56 mL) and water (1.56 mL), and stirred at 100° C. for 12 hrs. The reaction solution was adjusted to pH 6-7 with 1N HCl, extracted with EtOAc (10mL×3), and the organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated. The crude product was subjected to Pre-TLC (DCM: MeOH = 20:1) Separation and purification to obtain 93 mg of compound 4-3 as a yellow solid, with a yield of 69.41%.
[M+H +]:605.29。 [M+H + ]:605.29.
步骤3：化合物4的制备Step 3: Preparation of compound 4
0℃下，向95mg化合物4-3的二氧六环(3mL)溶液中滴加盐酸的甲醇溶液(2M,0.5mL)，25℃下搅拌1hr。反应液用饱和NaHCO 3水溶液调节pH为8-9淬灭反应，用EtOAc:THF＝5:1(10mL×3)萃取，合并有机相，有机相用饱和食盐水洗涤，无水硫酸钠干燥后浓缩，粗品经Pre-TLC(DCM:MeOH＝20:1)分离纯化后接着用Pre-HPLC(碱性)分离纯化，得到36mg化合物4为黄色固体，产率为45.32％。 At 0°C, a methanol solution of hydrochloric acid (2M, 0.5 mL) was added dropwise to 95 mg of compound 4-3 in dioxane (3 mL), and the mixture was stirred at 25°C for 1 hr. The reaction solution was quenched by adjusting the pH to 8-9 with saturated aqueous NaHCO 3 solution, extracted with EtOAc:THF=5:1 (10 mL×3), combined the organic phases, washed with saturated brine, and dried over anhydrous sodium sulfate After concentration, the crude product was separated and purified by Pre-TLC (DCM:MeOH=20:1) and then separated and purified by Pre-HPLC (basic) to obtain 36 mg of compound 4 as a yellow solid with a yield of 45.32%.
[M+H +]＝500.11 [M+H + ]=500.11
1H NMR(500MHz,CDCl 3):δ8.20(s,1H),7.35-7.33(dd,J＝8.0,1.5Hz,1H),7.32(s,1H),7.25-7.19(m,4H),7.11(t,J＝8.0Hz,1H),6.99(dd,J＝8.0,1.5Hz,1H),5.40(s,1H),4.06-3.93(m,3H),3.39-3.29(m,2H),3.11(d,J＝16.0Hz,1H),2.74(d,J＝16.0Hz,1H),1.93-1.87(m,1H),1.83-1.78(m,1H),1.63-1.59(m,1H),1.38-1.35(m,1H)。 1 H NMR (500MHz, CDCl 3 ): δ8.20 (s, 1H), 7.35-7.33 (dd, J = 8.0, 1.5Hz, 1H), 7.32 (s, 1H), 7.25-7.19 (m, 4H) ,7.11(t,J=8.0Hz,1H),6.99(dd,J=8.0,1.5Hz,1H),5.40(s,1H),4.06-3.93(m,3H),3.39-3.29(m,2H ), 3.11(d,J=16.0Hz,1H),2.74(d,J=16.0Hz,1H),1.93-1.87(m,1H),1.83-1.78(m,1H),1.63-1.59(m, 1H), 1.38-1.35 (m, 1H).
实施例5：化合物(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲酰胺的制备Example 5: Compound (S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazine-2-carboxamide
Figure PCTCN2020072774-appb-000027
Figure PCTCN2020072774-appb-000027
步骤1：化合物5-1的制备Step 1: Preparation of compound 5-1
将121mg化合物3-3溶于1.2mL 1,4-二氧六环和1.2mL水中，加入0.12mL的2.5M NaOH水溶液。反应液加热至100℃反应6hrs。TLC检测反应完全，加入10mL水和10mL的EtOAc萃取三次，合并有机层，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得32mg黄色固体的化合物5-1。Dissolve 121 mg of compound 3-3 in 1.2 mL of 1,4-dioxane and 1.2 mL of water, and add 0.12 mL of 2.5M NaOH aqueous solution. The reaction solution was heated to 100°C for 6hrs. TLC detected the completion of the reaction, added 10 mL of water and 10 mL of EtOAc for extraction three times, combined the organic layers, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 32 mg of yellow solid compound 5-1.
步骤2：化合物5的制备Step 2: Preparation of compound 5
RT条件下，将32mg化合物5-1溶于1.0mL的1,4-二氧六环中，滴加0.2mL的2M HCl的甲醇溶液，RT反应30mins。反应完全后旋干溶剂，加入1mL水，饱和的NaHCO 3水溶液调至pH为8。析出固体后加入10mL水，用EtOAc(10mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得16mg黄色固体的化合物5。 Under RT conditions, 32 mg of compound 5-1 was dissolved in 1.0 mL of 1,4-dioxane, 0.2 mL of 2M HCl in methanol was added dropwise, and the reaction was conducted at RT for 30 mins. After the reaction was complete, the solvent was spin-dried, 1 mL of water was added, and the pH was adjusted to 8 with saturated aqueous NaHCO 3 solution. After the solid precipitated out, 10 mL of water was added, extracted with EtOAc (10 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 16 mg of compound 5 as a yellow solid.
[M+H +]＝482.19。 [M+H + ]=482.19.
1H NMR(500MHz,DMSO-d 6):δ8.34(s,1H),8.04(s,1H),7.69-7.67(m,2H),7.32(d,J＝6.5Hz,1H),7.19-7.17(m,3H),6.36(s,2H),5.89(d,J＝5.0Hz,1H),4.01(t,J＝11.5Hz,2H),3.87(s,1H),3.28-3.23(m,2H),3.08(d,J＝15.5Hz,1H),2.66(d,J＝15.5Hz,1H),1.83-1.70(m,2H),1.53-1.51(m,1H),1.16-1.14(m,1H)。 1 H NMR (500MHz, DMSO-d 6 ): δ8.34(s,1H), 8.04(s,1H), 7.69-7.67(m,2H), 7.32(d,J=6.5Hz,1H), 7.19 -7.17 (m, 3H), 6.36 (s, 2H), 5.89 (d, J = 5.0 Hz, 1H), 4.01 (t, J = 11.5 Hz, 2H), 3.87 (s, 1H), 3.28-3.23 ( m,2H),3.08(d,J=15.5Hz,1H),2.66(d,J=15.5Hz,1H),1.83-1.70(m,2H),1.53-1.51(m,1H),1.16-1.14 (m,1H).
经由不同的反应起始原料和合适的试剂，采用与前述实施例5类似的方法制备表2中的化合物7-18、58和59。The compounds 7-18, 58 and 59 in Table 2 were prepared by using different reaction starting materials and appropriate reagents using a method similar to that of the foregoing Example 5.
表2Table 2
Figure PCTCN2020072774-appb-000028
Figure PCTCN2020072774-appb-000028
Figure PCTCN2020072774-appb-000029
Figure PCTCN2020072774-appb-000029
化合物7、8、9、10、11、12、14和59的核磁数据如下：The NMR data of compounds 7, 8, 9, 10, 11, 12, 14 and 59 are as follows:
1H NMR(500MHz,DMSO-d 6):δ8.39(s,1H),8.06(s,1H),7.92(d,J＝5.5Hz,1H),7.69(s,1H),7.31-7.30(m,1H),7.19-7.16(m,3H),6.39(d,J＝5.5Hz,1H),4.07-3.99(m,2H),3.93(s,3H),3.85(s,1H),3.29-3.24(m,2H),3.07(d,J＝15.4Hz,1H),2.64(d,J＝15.4Hz,1H),1.83-1.79(m,1H),1.74-1.70(m,1H),1.55-1.52(m,1H),1.15-1.13(m,1H)。(化合物7) 1 H NMR (500MHz, DMSO-d 6 ): δ8.39(s,1H), 8.06(s,1H), 7.92(d,J=5.5Hz,1H), 7.69(s,1H), 7.31-7.30 (m,1H),7.19-7.16(m,3H),6.39(d,J=5.5Hz,1H),4.07-3.99(m,2H),3.93(s,3H),3.85(s,1H), 3.29-3.24(m,2H),3.07(d,J=15.4Hz,1H), 2.64(d,J=15.4Hz,1H),1.83-1.79(m,1H),1.74-1.70(m,1H) ,1.55-1.52(m,1H),1.15-1.13(m,1H). (Compound 7)
1H NMR(500MHz,DMSO-d 6):δ8.55(d,J＝4.5Hz,1H),8.33(s,1H),8.01(s,1H),7.71-7.67(m,2H),7.63-7.62(m,1H),7.30(d,J＝6.5Hz,1H),7.17-7.14(m,3H),4.01-3.97(m,2H),3.84(s,1H),3.28-3.23(m,2H),3.05(d,J＝15.0Hz,1H),2.63(d,J＝14.5Hz,1H),1.80-1.77(m,1H),1.70-1.67(m,1H),1.52-1.50(m,1H),1.13-1.10(m,1H)。(化合物8) 1 H NMR (500MHz, DMSO-d 6 ): δ8.55 (d, J = 4.5Hz, 1H), 8.33 (s, 1H), 8.01 (s, 1H), 7.71-7.67 (m, 2H), 7.63 -7.62(m,1H),7.30(d,J=6.5Hz,1H),7.17-7.14(m,3H),4.01-3.97(m,2H),3.84(s,1H),3.28-3.23(m ,2H),3.05(d,J=15.0Hz,1H),2.63(d,J=14.5Hz,1H),1.80-1.77(m,1H),1.70-1.67(m,1H),1.52-1.50( m, 1H), 1.13-1.10 (m, 1H). (Compound 8)
1H NMR(500MHz,CDCl 3):δ8.55(dd,J＝4.5,1.0Hz,1H),8.22(s,1H),7.67(d,J＝7.5Hz,1H),7.39(dd,J＝8.2,4.5Hz,1H),7.12(br,1H),7.10(d,J＝8.3Hz,1H),6.89(d,J＝2.5Hz,1H),6.76(dd,J＝8.2,2.5Hz,1H),5.36(br,1H),4.03-4.98(m,2H),3.95(s,1H),3.81(s,3H),3.38-3.27(m,2H),3.03(d,J＝15.0Hz,1H),2.67(d,J＝15.0Hz,1H),1.92-1.86(m,1H),1.80-1.74(m,1H),1.62-1.59(m,1H),1.36-1.33(m,1H)。(化合物9) 1 H NMR (500MHz, CDCl 3 ): δ8.55 (dd, J = 4.5, 1.0 Hz, 1H), 8.22 (s, 1H), 7.67 (d, J = 7.5 Hz, 1H), 7.39 (dd, J ＝8.2,4.5Hz,1H),7.12(br,1H),7.10(d,J＝8.3Hz,1H), 6.89(d,J＝2.5Hz,1H), 6.76(dd,J＝8.2,2.5Hz ,1H),5.36(br,1H),4.03-4.98(m,2H),3.95(s,1H),3.81(s,3H),3.38-3.27(m,2H),3.03(d,J=15.0 Hz,1H),2.67(d,J=15.0Hz,1H),1.92-1.86(m,1H),1.80-1.74(m,1H),1.62-1.59(m,1H),1.36-1.33(m, 1H). (Compound 9)
1H NMR(500MHz,DMSO-d 6):δ8.36(s,1H),8.04(s,1H),7.69(m,2H),7.38-7.13(m,3H),6.38(s,2H),5.89(d,J＝5.0Hz,1H),4.03-3.97(m,2H),3.94(s,1H),3.32-3.24(m,2H),3.04(d,J＝17.0Hz,1H),2.65(d,J＝17.0Hz,1H),1.82-1.72(m,2H),1.57-1.54(m,1H),1.16-1.14(m,1H)。(化合物10) 1 H NMR (500MHz, DMSO-d 6 ): δ8.36 (s, 1H), 8.04 (s, 1H), 7.69 (m, 2H), 7.38-7.13 (m, 3H), 6.38 (s, 2H) ,5.89(d,J=5.0Hz,1H),4.03-3.97(m,2H),3.94(s,1H),3.32-3.24(m,2H),3.04(d,J=17.0Hz,1H), 2.65 (d, J=17.0 Hz, 1H), 1.82-1.72 (m, 2H), 1.57-1.54 (m, 1H), 1.16-1.14 (m, 1H). (Compound 10)
1H NMR(500MHz,DMSO-d 6):δ8.35(s,1H),8.05(s,1H),7.69(m,2H),7.19(t,J＝7.5Hz,1H),6.95(d,J＝7.5Hz,1H),6.82(d,J＝7.5Hz,1H),6.38(s,2H),5.88(d,J＝5.0Hz,1H),4.05-3.97(m,2H),3.91(s,1H),3.77(s,3H),3.27-3.24(m,2H),2.96(d,J＝16.0Hz,1H),2.58(d,J＝16.0Hz,1H),1.79-1.75(m,2H),1.49-1.51(m,1H),1.23-1.21(m,1H)。(化合物11) 1 H NMR (500MHz, DMSO-d 6 ): δ8.35(s,1H), 8.05(s,1H), 7.69(m,2H), 7.19(t,J=7.5Hz,1H), 6.95(d ,J=7.5Hz,1H),6.82(d,J=7.5Hz,1H),6.38(s,2H),5.88(d,J=5.0Hz,1H),4.05-3.97(m,2H),3.91 (s, 1H), 3.77 (s, 3H), 3.27-3.24 (m, 2H), 2.96 (d, J = 16.0 Hz, 1H), 2.58 (d, J = 16.0 Hz, 1H), 1.79-1.75 ( m, 2H), 1.49-1.51 (m, 1H), 1.23-1.21 (m, 1H). (Compound 11)
1H NMR(500MHz,CDCl 3):δ8.28(s,1H),7.75(d,J＝5.0Hz,1H),7.12(d,J＝8.5Hz,1H),6.89(d,J＝2.0Hz,1H),6.77(dd,J＝8.5Hz,J＝2.0Hz,1H),6.07(d,J＝5.5Hz,1H),4.90(s,2H),4.08-4.03(m,2H),3.96(s,1H),3.81(s,3H),3.41-3.31(m,2H),3.04(d,J＝15.0Hz,1H),2.68(d,J＝15.0Hz,1H),1.94-1.88(m,1H),1.82-1.77(m,1H),1.64-1.62(m,1H),1.38-1.35(m,1H)。(化合物12) 1 H NMR(500MHz, CDCl 3 ): δ8.28(s,1H), 7.75(d,J=5.0Hz,1H), 7.12(d,J=8.5Hz,1H), 6.89(d,J=2.0 Hz, 1H), 6.77 (dd, J = 8.5 Hz, J = 2.0 Hz, 1H), 6.07 (d, J = 5.5 Hz, 1H), 4.90 (s, 2H), 4.08-4.03 (m, 2H), 3.96(s,1H),3.81(s,3H),3.41-3.31(m,2H),3.04(d,J=15.0Hz,1H), 2.68(d,J=15.0Hz,1H),1.94-1.88 (m, 1H), 1.82-1.77 (m, 1H), 1.64-1.62 (m, 1H), 1.38-1.35 (m, 1H). (Compound 12)
1H NMR(500MHz,CDCl 3):δ8.29(s,1H),7.74(d,J＝5.4Hz,1H),7.60(s,1H),7.49(d,J＝7.8Hz,1H),7.32(d,J＝7.8Hz,1H),6.07(d,J＝5.4Hz,1H),4.16-4.04(m,2H),4.03(s,1H),3.43-3.29(m,2H),3.18(d,J＝16.0Hz,1H),2.79(d,J＝16.0Hz,1H),1.98-1.92(m,1H),1.85-1.80(m,1H),1.65-1.62(m,1H),1.35-1.31(m,1H)。(化合物14) 1 H NMR(500MHz, CDCl 3 ): δ8.29(s,1H), 7.74(d,J=5.4Hz,1H), 7.60(s,1H), 7.49(d,J=7.8Hz,1H), 7.32(d,J=7.8Hz,1H),6.07(d,J=5.4Hz,1H),4.16-4.04(m,2H),4.03(s,1H),3.43-3.29(m,2H),3.18 (d,J=16.0Hz,1H),2.79(d,J=16.0Hz,1H),1.98-1.92(m,1H),1.85-1.80(m,1H),1.65-1.62(m,1H), 1.35-1.31 (m, 1H). (Compound 14)
[M+H +]＝562.20。 [M+H + ]=562.20.
1H NMR(500MHz,DMSO-d 6)δ8.51(s,1H),8.38(s,1H),8.03(s,1H),7.78(d,J＝5.4Hz,1H),7.71-7.66(m,1H),7.36(d,J＝1.9Hz,1H),7.33-7.27(m,1H),7.21-7.13(m,3H),6.15(d,J＝5.4Hz,1H),6.09(d,J＝1.9Hz,1H),4.07-3.99(m,2H),3.84(s,1H),3.56(s,3H),3.30-3.22(m,2H),3.07(d,J＝15.6Hz,1H),2.64(d,J＝15.6Hz,1H),1.85-1.78(m,1H),1.76-1.68(m,1H),1.55-1.50(m,1H),1.17-1.11(m,1H).(化合物59) 1 H NMR(500MHz,DMSO-d 6 )δ8.51(s,1H), 8.38(s,1H), 8.03(s,1H), 7.78(d,J=5.4Hz,1H), 7.71-7.66( m,1H), 7.36(d,J=1.9Hz,1H),7.33-7.27(m,1H),7.21-7.13(m,3H),6.15(d,J=5.4Hz,1H), 6.09(d ,J=1.9Hz,1H),4.07-3.99(m,2H),3.84(s,1H),3.56(s,3H),3.30-3.22(m,2H),3.07(d,J=15.6Hz, 1H), 2.64(d,J=15.6Hz,1H),1.85-1.78(m,1H),1.76-1.68(m,1H),1.55-1.50(m,1H),1.17-1.11(m,1H) .(Compound 59)
中间体化合物M15-3的制备：Preparation of intermediate compound M15-3:
Figure PCTCN2020072774-appb-000030
Figure PCTCN2020072774-appb-000030
步骤1：化合物M15-2的制备Step 1: Preparation of compound M15-2
将530mg化合物M15-1、190mg化合物M1-2溶解于20mL二氧六环中，加入73mg Pd 2(dba) 3、92mg Xantphos和410mg DIEA。氮气置换三次，氮气保护下反应升至85℃搅拌反应12hrs。LCMS检测反应完全，将反应液过滤，向滤液中加入75mL饱和食盐水，用EtOAc(20mL×4)萃取，合并有机相，用无水硫酸钠干燥，减压浓缩，残余物经柱层析纯 化得510mg化合物M15-2。 530 mg of compound M15-1 and 190 mg of compound M1-2 were dissolved in 20 mL of dioxane, and 73 mg of Pd 2 (dba) 3 , 92 mg of Xantphos and 410 mg of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 85°C and stirred for 12 hrs. The reaction was completed by LCMS, the reaction solution was filtered, 75mL saturated brine was added to the filtrate, extracted with EtOAc (20mL×4), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography 510 mg of compound M15-2 was obtained.
步骤2：化合物M15-3的制备Step 2: Preparation of compound M15-3
将250mg化合物M15-2溶解于3mL THF中，RT下滴加EtONa(1.4mL，20％的EtOH溶液)，RT搅拌反应45mins。LCMS检测反应完全，减压浓缩，加入5mL的DCM超声5mins，静置后倾去上清液，加入5mL MTBE超声5分钟，静置后倾去上清液，干燥得固体116mg化合物M15-3。250 mg of compound M15-2 was dissolved in 3 mL of THF, EtONa (1.4 mL, 20% EtOH solution) was added dropwise at RT, and the reaction was stirred at RT for 45 mins. LCMS detects that the reaction is complete, concentrates under reduced pressure, adds 5 mL of DCM and sonicates for 5 mins, then decanted the supernatant after standing, adds 5 mL of MTBE and sonicates for 5 minutes, after standing, decanted the supernatant, and dried to obtain 116 mg of compound M15-3 as a solid.
中间体化合物M16-5的制备：Preparation of intermediate compound M16-5:
Figure PCTCN2020072774-appb-000031
Figure PCTCN2020072774-appb-000031
步骤1：化合物M16-3的制备Step 1: Preparation of compound M16-3
将1.90g NaH(含量60％)分批加入25mL THF氮气保护下反应升至60℃，滴加5.00g化合物M16-2和3.46g化合物M16-1的THF(40mL)溶液，搅拌反应2.5hrs。TLC检测反应完全，将反应液倒入20mL水中，加DCM萃取(40mL×2)，合并有机相，用30mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得4.45g黄色固体物化合物M16-3。Add 1.90g of NaH (content 60%) to 25mL of THF in batches to raise the temperature to 60°C under nitrogen protection, add dropwise a THF (40mL) solution of 5.00g of compound M16-2 and 3.46g of compound M16-1, and stir the reaction for 2.5hrs. TLC detected the completion of the reaction. The reaction solution was poured into 20 mL of water, extracted with DCM (40 mL×2), and the organic phases were combined, washed with 30 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography. 4.45g yellow solid compound M16-3.
步骤2：化合物M16-4的制备Step 2: Preparation of compound M16-4
将4.45g化合物M16-3和2.05g化合物M1-2溶解于50mL二氧六环中，加入283mg Pd 2(dba) 3、448mg Xantphos和4.00g DIPEA，氮气置换三次，反应液升至85℃搅拌反应12hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(20mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得5.38g化合物M16-4。 Dissolve 4.45g compound M16-3 and 2.05g compound M1-2 in 50mL dioxane, add 283mg Pd 2 (dba) 3 , 448mg Xantphos and 4.00g DIPEA, replace with nitrogen three times, and raise the reaction solution to 85℃ and stir Reaction for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (20 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 5.38 g of compound M16-4.
步骤3：化合物M16-5的制备Step 3: Preparation of compound M16-5
将5.38g化合物M16-4溶解于50mL THF中，-30℃下滴加EtONa(9mL，20％的EtOH溶液)，RT搅拌反应2hrs。TLC检测反应完全，减压浓缩，加入30mL DCM搅拌30mins，反应液过滤，滤饼用DCM(15mL×2)洗涤，抽干得4.29g化合物M16-5。5.38g of compound M16-4 was dissolved in 50mL THF, EtONa (9mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 2hrs. TLC detected the completion of the reaction, concentrated under reduced pressure, added 30 mL DCM and stirred for 30 mins, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), and drained to obtain 4.29 g of compound M16-5.
中间体化合物M17-3的制备：Preparation of intermediate compound M17-3:
Figure PCTCN2020072774-appb-000032
Figure PCTCN2020072774-appb-000032
步骤1：化合物M17-1的制备Step 1: Preparation of compound M17-1
将700mg化合物M17-SM和2.5mL NH(CH 3) 2水溶液溶解于2.5mL二氧六环中，氮气置换三次，氮气保护下反应升至90℃搅拌反应2hr。TLC检测反应完全，加入10mL水和10mL乙酸乙酯萃取，分离有机相，用10mL饱和氯化钠溶液洗涤，无水硫酸钠干燥。旋出溶剂，得到708mg化合物M17-1。 700 mg of compound M17-SM and 2.5 mL of NH(CH 3 ) 2 aqueous solution were dissolved in 2.5 mL of dioxane, replaced with nitrogen three times, and the reaction was raised to 90° C. under the protection of nitrogen and stirred for 2 hours. TLC detects that the reaction is complete, 10 mL of water and 10 mL of ethyl acetate are added for extraction, the organic phase is separated, washed with 10 mL of saturated sodium chloride solution, and dried with anhydrous sodium sulfate. The solvent was spun off to obtain 708 mg of compound M17-1.
步骤2：化合物M17-2的制备Step 2: Preparation of compound M17-2
将708mg化合物M17-1和320mg化合物M1-2溶解于10mL二氧六环中，加入243mg Pd 2(dba) 3、308mg Xantphos和688mg DIEA。氮气置换三次，氮气保护下反应升至90℃搅拌反应5hr。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，经柱层析纯化得584mg化合物M17-2。 708 mg of compound M17-1 and 320 mg of compound M1-2 were dissolved in 10 mL of dioxane, and 243 mg of Pd 2 (dba) 3 , 308 mg of Xantphos and 688 mg of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 5 hours. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 584 mg of compound M17-2.
步骤3：化合物M17-3的制备Step 3: Preparation of compound M17-3
将584mg化合物M17-2溶解于6mL THF中，室温下滴加EtONa(185mg)与CH 3ONa(183mg)的乙醇溶液(4mL)，RT搅拌反应2hr。TLC检测反应完全，减压浓缩得到固体，加入10mL的甲基叔丁基醚搅拌5min，除去反应液，重复三次，抽干，得固体500mg粗品化合物M17-3，可直接用于下一步反应。 584 mg of compound M17-2 was dissolved in 6 mL of THF, and an ethanol solution (4 mL) of EtONa (185 mg) and CH 3 ONa (183 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 500 mg of crude compound M17-3, which could be used directly in the next reaction.
中间体化合物M18-3的制备：Preparation of intermediate compound M18-3:
Figure PCTCN2020072774-appb-000033
Figure PCTCN2020072774-appb-000033
步骤1：化合物M18-1的制备Step 1: Preparation of compound M18-1
将700mg化合物M17-SM和2.5mL甲胺水溶液溶解于2.5mL二氧六环中，氮气置换三次，氮气保护下反应升至90℃搅拌反应2hr。TLC检测反应完全，加入10mL水和10mL乙酸乙酯萃取，分离有机相，用10mL饱和氯化钠溶液洗涤，无水硫酸钠干燥。旋出溶剂，得到728mg化合物M18-1。700 mg of compound M17-SM and 2.5 mL of methylamine aqueous solution were dissolved in 2.5 mL of dioxane, and replaced with nitrogen three times. Under the protection of nitrogen, the reaction was raised to 90° C. and stirred for 2 hr. TLC detects that the reaction is complete, 10 mL of water and 10 mL of ethyl acetate are added for extraction, the organic phase is separated, washed with 10 mL of saturated sodium chloride solution, and dried with anhydrous sodium sulfate. The solvent was spun off to obtain 728 mg of compound M18-1.
步骤2：化合物M18-2的制备Step 2: Preparation of compound M18-2
将728mg化合物M18-1和347mg化合物M1-2溶解于7mL二氧六环中，加入264 mg Pd 2(dba) 3、334mg Xantphos和747mg DIEA。氮气置换三次，氮气保护下反应升至90℃搅拌反应5hr。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，经柱层析纯化得646mg化合物M18-2。 728 mg of compound M18-1 and 347 mg of compound M1-2 were dissolved in 7 mL of dioxane, and 264 mg of Pd 2 (dba) 3 , 334 mg of Xantphos and 747 mg of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 5 hours. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 646 mg of compound M18-2.
步骤3：化合物M18-3的制备Step 3: Preparation of compound M18-3
将646mg化合物M18-2溶解于7mL THF中，室温下滴加EtONa(180mg)与CH 3ONa(243mg)的乙醇溶液(3mL)，RT搅拌反应2hr。TLC检测反应完全，减压浓缩得到固体，加入10mL的甲基叔丁基醚搅拌5min，除去反应液，重复三次，抽干，得固体500mg粗品化合物M18-3，可直接用于下一步反应。 646 mg of compound M18-2 was dissolved in 7 mL of THF, and an ethanol solution (3 mL) of EtONa (180 mg) and CH 3 ONa (243 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 500 mg of crude compound M18-3, which can be directly used in the next reaction.
中间体化合物M19-2的制备：Preparation of intermediate compound M19-2:
Figure PCTCN2020072774-appb-000034
Figure PCTCN2020072774-appb-000034
步骤1：化合物M19-1的制备Step 1: Preparation of compound M19-1
将1g化合物M19-SM和578mg化合物M1-2溶解于10mL二氧六环中，加入220mg Pd 2(dba) 3、278mg Xantphos和1.24g DIEA。氮气置换三次，氮气保护下反应升至90℃搅拌反应5hr。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，经柱层析纯化得885mg化合物M19-1。 1 g of compound M19-SM and 578 mg of compound M1-2 were dissolved in 10 mL of dioxane, and 220 mg of Pd 2 (dba) 3 , 278 mg of Xantphos and 1.24 g of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 5 hours. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 885 mg of compound M19-1.
步骤2：化合物M19-2的制备Step 2: Preparation of compound M19-2
将885mg化合物M19-1溶解于8mL THF中，室温下滴加EtONa(268mg)与CH 3ONa(251mg)的乙醇溶液(4mL)，RT搅拌反应2hr。TLC检测反应完全，减压浓缩得到固体，加入10mL的甲基叔丁基醚搅拌5min，除去反应液，重复三次，抽干，得固体700mg粗品化合物M19-2，可直接用于下一步反应。 885 mg of compound M19-1 was dissolved in 8 mL of THF, and an ethanol solution (4 mL) of EtONa (268 mg) and CH 3 ONa (251 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 700 mg of crude compound M19-2, which can be used directly in the next reaction.
中间体化合物M20-3的制备：Preparation of intermediate compound M20-3:
Figure PCTCN2020072774-appb-000035
Figure PCTCN2020072774-appb-000035
步骤1：化合物M20-1的制备Step 1: Preparation of compound M20-1
将1g化合物M20-SM和3mL NH(CH 3) 2水溶液溶解于3mL二氧六环中，氮气置换三次，氮气保护下反应升至90℃搅拌反应2hr。TLC检测反应完全，加入10mL水和10 mL乙酸乙酯萃取，分离有机相，用10mL饱和氯化钠溶液洗涤，无水硫酸钠干燥。旋出溶剂，得到1.03g化合物M20-1。 1 g of compound M20-SM and 3 mL of NH(CH 3 ) 2 aqueous solution were dissolved in 3 mL of dioxane, replaced with nitrogen three times, and the reaction was raised to 90° C. under the protection of nitrogen and stirred for 2 hours. TLC detects the completion of the reaction, adds 10 mL of water and 10 mL of ethyl acetate for extraction, separates the organic phase, washes with 10 mL of saturated sodium chloride solution, and dried with anhydrous sodium sulfate. The solvent was spun off to obtain 1.03 g of compound M20-1.
步骤2：化合物M20-2的制备Step 2: Preparation of compound M20-2
将1.03g化合物M20-1和438mg化合物M1-2溶解于10mL二氧六环中，加入334mg Pd 2(dba) 3、422mg Xantphos和942mg DIEA。氮气置换三次，氮气保护下反应升至90℃搅拌反应4hr。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，经柱层析纯化得968mg化合物M20-2。 1.03 g of compound M20-1 and 438 mg of compound M1-2 were dissolved in 10 mL of dioxane, and 334 mg of Pd 2 (dba) 3 , 422 mg of Xantphos and 942 mg of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 4 hours. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 968 mg of compound M20-2.
步骤3：化合物M20-3的制备Step 3: Preparation of compound M20-3
将968mg化合物M20-2溶解于10mL THF中，室温下滴加EtONa(120mg)与CH 3ONa(228mg)的乙醇溶液(3mL)，RT搅拌反应2hr。TLC检测反应完全，减压浓缩得到固体，加入10mL的甲基叔丁基醚搅拌5min，除去反应液，重复三次，抽干，得固体900mg粗品化合物M20-3，可直接用于下一步反应。 968 mg of compound M20-2 was dissolved in 10 mL of THF, and the ethanol solution (3 mL) of EtONa (120 mg) and CH 3 ONa (228 mg) was added dropwise at room temperature, and the reaction was stirred at RT for 2 hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, added 10 mL of methyl tert-butyl ether and stirred for 5 min, removed the reaction liquid, repeated three times, and drained to obtain a solid 900 mg of crude compound M20-3, which can be directly used in the next reaction.
中间体化合物M21-3的制备：Preparation of intermediate compound M21-3:
Figure PCTCN2020072774-appb-000036
Figure PCTCN2020072774-appb-000036
步骤1：化合物M21-1的制备Step 1: Preparation of compound M21-1
将4.0g化合物M17-SM溶解于20mL 25％氨水溶液和20mL二氧六环中，封管70℃搅拌反应48hr。TLC检测反应完全，反应液加入水(20ml)，EA(50ml×2)萃取，有机相饱和盐水洗涤(10ml×2)，无水硫酸钠干燥，旋干后固体用正己烷打浆，过滤得3.67g，即化合物M21-1。4.0 g of compound M17-SM was dissolved in 20 mL 25% ammonia solution and 20 mL dioxane, and the tube was sealed at 70° C. and the reaction was stirred for 48 hours. TLC detects that the reaction is complete, the reaction solution is added with water (20ml), extracted with EA (50ml×2), the organic phase is washed with saturated brine (10ml×2), dried with anhydrous sodium sulfate, spin-dried, the solid is slurried with n-hexane and filtered to obtain 3.67 g, Compound M21-1.
步骤2：化合物M21-2的制备Step 2: Preparation of compound M21-2
将2.92g化合物M21-1和1.47g化合物M1-2溶解于45mL二氧六环中，加入354mg Pd 2(dba) 3、225mg Xantphos和3.17g DIEA。氮气置换三次，氮气保护下反应升至80℃搅拌反应过夜。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，经柱层析纯化得2.68g化合物M21-2。 2.92 g of compound M21-1 and 1.47 g of compound M1-2 were dissolved in 45 mL of dioxane, and 354 mg of Pd 2 (dba) 3 , 225 mg of Xantphos and 3.17 g of DIEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 80°C and stirred overnight. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and purified by column chromatography to obtain 2.68 g of compound M21-2.
步骤3：化合物M21-3的制备Step 3: Preparation of compound M21-3
将2.68g化合物M21-2溶解于40mL THF中，降温-30℃滴加EtONa(1.19g)的乙醇(12mL)溶液，然后自然升温RT反应3hr。TLC检测反应完全，减压浓缩得到固体，用 MTBE(20mL)打浆，得固体2.12g粗品化合物M21-3，可直接用于下一步反应。Dissolve 2.68g of compound M21-2 in 40mL THF, add dropwise EtONa (1.19g) ethanol (12mL) solution at a temperature of -30°C, and then naturally increase RT to react for 3hr. TLC detected that the reaction was complete, concentrated under reduced pressure to obtain a solid, which was beaten with MTBE (20 mL) to obtain 2.12 g of crude compound M21-3 as a solid, which could be used directly in the next reaction.
中间体M22-2和M23-1的制备：Preparation of intermediates M22-2 and M23-1:
Figure PCTCN2020072774-appb-000037
Figure PCTCN2020072774-appb-000037
步骤1：化合物M22-1的制备Step 1: Preparation of compound M22-1
将1.29g化合物M22-SM和1.60g化合物M6溶解于20mL四氢呋喃中，滴加1.12g DIEA，60℃搅拌反应12小时。TLC检测反应完全，减压浓缩，加入50mL水，再用EtOAc(50mL×3)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得1.60g化合物M22-1。Dissolve 1.29 g of compound M22-SM and 1.60 g of compound M6 in 20 mL of tetrahydrofuran, add 1.12 g DIEA dropwise, and stir and react at 60°C for 12 hours. TLC detected the completion of the reaction, concentrated under reduced pressure, added 50 mL of water, and extracted with EtOAc (50 mL×3), combined the organic layers, dried over sodium sulfate and desolventized, purified by column chromatography to obtain 1.60 g of compound M22-1.
步骤2：化合物M22-2和M23-1的制备Step 2: Preparation of compounds M22-2 and M23-1
氮气保护下，将1.00g化合物M22-1溶解于30mL无水二氯甲烷中，温度降至-78℃，滴加DIBAL-H(1M，9.6mL)的正己烷溶液。-78℃搅拌反应1小时。再缓慢升温至-40℃，继续反应2小时。TLC检测原料反应完全，在0℃下，缓慢滴加0.4mL水，再滴加氢氧化钠水溶液(15％，0.4mL)，再加入1mL水。升至室温搅拌15分钟。加入硫酸钠干燥，搅拌十分钟后过滤。滤液浓缩，经柱层析纯化得0.40g化合物M22-2和0.11g化合物M23-1。Under nitrogen protection, 1.00 g of compound M22-1 was dissolved in 30 mL of anhydrous dichloromethane, the temperature was reduced to -78°C, and DIBAL-H (1M, 9.6 mL) in n-hexane was added dropwise. The reaction was stirred at -78°C for 1 hour. Then slowly increase the temperature to -40°C and continue the reaction for 2 hours. TLC detected that the reaction of the raw materials was complete. At 0° C., 0.4 mL of water was slowly added dropwise, and then sodium hydroxide aqueous solution (15%, 0.4 mL) was added dropwise, and then 1 mL of water was added. Warm to room temperature and stir for 15 minutes. Add sodium sulfate to dry, stir for ten minutes and filter. The filtrate was concentrated and purified by column chromatography to obtain 0.40 g of compound M22-2 and 0.11 g of compound M23-1.
中间体化合物M24-4的制备：Preparation of intermediate compound M24-4:
Figure PCTCN2020072774-appb-000038
Figure PCTCN2020072774-appb-000038
步骤1：化合物M24-2的制备Step 1: Preparation of compound M24-2
将1.00g化合物M24-1溶解于10mL无水乙腈中，加入1.45g碳酸钾，85℃搅拌反应2小时。TLC检测反应完全，减压除去溶剂，加入50mL水，再用EA(50mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得710mg化合物M24-2。Dissolve 1.00 g of compound M24-1 in 10 mL of anhydrous acetonitrile, add 1.45 g of potassium carbonate, and stir and react at 85°C for 2 hours. TLC detected that the reaction was complete, the solvent was removed under reduced pressure, 50 mL of water was added, and then extracted with EA (50 mL×2), and the organic layers were combined, dried over sodium sulfate and desolventized, and purified by column chromatography to obtain 710 mg of compound M24-2.
步骤2：化合物M24-3的制备Step 2: Preparation of compound M24-3
将710mg化合物M24-2、455mg化合物M1-2、895mg DIEA、63mg Pd 2(dba) 3和100mg Xantphos溶解于10mL二氧六环中，氮气置换3次后，85℃反应12hrs。停止加热，减压浓缩，加入30mL水，再用EA(30mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得84mg化合物M24-3。 710 mg of compound M24-2, 455 mg of compound M1-2, 895 mg of DIEA, 63 mg of Pd 2 (dba) 3 and 100 mg of Xantphos were dissolved in 10 mL of dioxane, replaced with nitrogen three times, and reacted at 85°C for 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and then extract with EA (30 mL×2), combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 84 mg of compound M24-3.
步骤3：化合物M24-4的制备Step 3: Preparation of compound M24-4
氮气保护下，将84mg化合物M24-3溶于2mL无水四氢呋喃中，将EtONa的EtOH溶液(0.85mL，20％)缓慢滴加入上述溶液中，剧烈搅拌40min。TLC检测反应完全，将反应液减压浓缩，残余物加入5mL甲基叔丁基醚打浆30min，过滤，取滤饼真空干燥得60mg棕色固体M24-4。Under nitrogen protection, 84 mg of compound M24-3 was dissolved in 2 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (0.85 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was slurried by adding 5 mL of methyl tert-butyl ether for 30 min, filtered, and the filter cake was taken and dried under vacuum to obtain 60 mg of brown solid M24-4.
中间体化合物M25-5的制备：Preparation of intermediate compound M25-5:
Figure PCTCN2020072774-appb-000039
Figure PCTCN2020072774-appb-000039
步骤1：化合物M25-2的制备Step 1: Preparation of compound M25-2
0℃下，将氢氧化钠(546mg)水溶液(25mL)，滴加到溶有1.00g化合物M25-1、2.60g碘单质和1.70g碘化钾的水溶液中。常温反应2h，TLC检测反应完全。用饱和氯化铵溶液调节pH至中性。再用硫代硫酸钠溶液除去碘单质。用甲基叔丁基醚(50mL×2)萃取，合并有机层后，用硫酸钠干燥、脱溶。得到1.45g化合物M25-2。At 0°C, an aqueous solution (25 mL) of sodium hydroxide (546 mg) was added dropwise to an aqueous solution in which 1.00 g of compound M25-1, 2.60 g of elemental iodine and 1.70 g of potassium iodide were dissolved. The reaction was carried out at room temperature for 2 hours, and the reaction was completed as detected by TLC. Adjust the pH to neutral with saturated ammonium chloride solution. Then use sodium thiosulfate solution to remove elemental iodine. It was extracted with methyl tert-butyl ether (50 mL×2), and the organic layers were combined, dried with sodium sulfate and desolvated. 1.45 g of compound M25-2 was obtained.
步骤2：化合物M25-3的制备Step 2: Preparation of compound M25-3
将1.45g化合物M25-2、1.13g碘甲烷和1.10碳酸钾溶于DMF(20mL)中，常温反应3h，TLC检测反应完全。用甲基叔丁基醚(50mL×2)萃取，合并有机层后，用硫酸钠干燥、脱溶。经柱层析纯化得1.19g化合物M25-2。1.45g of compound M25-2, 1.13g of methyl iodide and 1.10 potassium carbonate were dissolved in DMF (20mL), reacted at room temperature for 3h, TLC detected that the reaction was complete. It was extracted with methyl tert-butyl ether (50 mL×2), and the organic layers were combined, dried with sodium sulfate and desolvated. Purified by column chromatography to obtain 1.19 g of compound M25-2.
步骤3：化合物M25-4的制备Step 3: Preparation of compound M25-4
将1.19g化合物M25-3、498mg化合物M1-2、1.07g DIEA、76mg Pd 2(dba) 3和120mg Xantphos溶解于10mL二氧六环中，氮气置换3次后，85℃反应12hrs。停止加热，减压浓缩，加入30mL水，再用EA(30mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得837mg化合物M25-4。 Dissolve 1.19 g of compound M25-3, 498 mg of compound M1-2, 1.07 g of DIEA, 76 mg of Pd 2 (dba) 3 and 120 mg of Xantphos in 10 mL of dioxane, replace with nitrogen three times, and react at 85°C for 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 837 mg of compound M25-4.
步骤4：化合物M25-5的制备Step 4: Preparation of compound M25-5
氮气保护下，将870mg化合物M25-4溶于8mL无水四氢呋喃中，将EtONa的EtOH溶液(0.46mL，20％)缓慢滴加入上述溶液中，剧烈搅拌40min。TLC检测反应完全，将反应液减压浓缩，残余物加入5mL甲基叔丁基醚打浆30min，过滤，取滤饼真空干燥得623mg棕色固体M25-5。Under nitrogen protection, 870 mg of compound M25-4 was dissolved in 8 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (0.46 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was slurried by adding 5 mL of methyl tert-butyl ether for 30 min, filtered, and the filter cake was vacuum-dried to obtain 623 mg of brown solid M25-5.
中间体化合物M26-3的制备Preparation of intermediate compound M26-3
Figure PCTCN2020072774-appb-000040
Figure PCTCN2020072774-appb-000040
步骤1：化合物M26-2的制备Step 1: Preparation of compound M26-2
将1.00g化合物M26-1、578mg化合物M1-2、1.24g DIEA、88mg Pd 2(dba) 3和139mg Xantphos溶解于10mL二氧六环中，氮气置换3次后，85℃反应12hrs。停止加热，减压浓缩，加入30mL水，再用EA(30mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得992mg化合物M26-2。 1.00 g of compound M26-1, 578 mg of compound M1-2, 1.24 g of DIEA, 88 mg of Pd 2 (dba) 3 and 139 mg of Xantphos were dissolved in 10 mL of dioxane, and after nitrogen replacement three times, the reaction was carried out at 85° C. for 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 992 mg of compound M26-2.
步骤2：化合物M26-3的制备Step 2: Preparation of compound M26-3
氮气保护下，将992mg化合物M26-2溶于10mL无水四氢呋喃中，将EtONa的EtOH溶液(1.5mL，20％)缓慢滴加入上述溶液中，剧烈搅拌40min。TLC检测反应完全，将反应液减压浓缩，残余物加入10mL甲基叔丁基醚打浆30min，过滤，取滤饼真空干燥得504mg棕色固体M26-3。Under nitrogen protection, 992 mg of compound M26-2 was dissolved in 10 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (1.5 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was slurried by adding 10 mL of methyl tert-butyl ether for 30 minutes, filtered, and the filter cake was vacuum-dried to obtain 504 mg of brown solid M26-3.
中间体化合物M27-11的制备：Preparation of intermediate compound M27-11:
Figure PCTCN2020072774-appb-000041
Figure PCTCN2020072774-appb-000041
步骤1：化合物M27-3的制备Step 1: Preparation of compound M27-3
将10.00g化合物M27-1和7.59g化合物M27-2溶解于170mL氯仿中，加入350mg碘单质，常温搅拌反应24小时。TLC检测反应完全，将化合物倒入硫代硫酸钠水溶液中(100mL，0.4mol/L)，再加入氢氧化钠水溶液(70mL,40％)，再用氯仿(150mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，用二氯甲烷/正己烷重结晶，得到8.77g化合物M27-3。10.00 g of compound M27-1 and 7.59 g of compound M27-2 were dissolved in 170 mL of chloroform, 350 mg of iodine was added, and the reaction was stirred at room temperature for 24 hours. TLC detects that the reaction is complete. Pour the compound into sodium thiosulfate aqueous solution (100mL, 0.4mol/L), then add sodium hydroxide aqueous solution (70mL, 40%), and then extract with chloroform (150mL×2), and combine the organic layers After drying with sodium sulfate, the solution was removed and recrystallized with dichloromethane/n-hexane to obtain 8.77g of compound M27-3.
步骤2：化合物M27-5的制备Step 2: Preparation of compound M27-5
将8.77g化合物M27-3溶解于300mL无水四氢呋喃中，降温至-30℃，滴加正丁基锂(2.5M，16mL)的正己烷溶液，保温搅拌1小时。随后将8.15g化合物M27-4溶在30 mL四氢呋喃中缓慢滴加到反应液中，滴加完毕后升温至-10℃继续反应2.5小时。反应液用200mL饱和食盐水淬灭，用EtOAc(300mL×2)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，经柱层析纯化得11.06g化合物M27-5。8.77 g of compound M27-3 was dissolved in 300 mL of anhydrous tetrahydrofuran, the temperature was lowered to -30° C., a n-hexane solution of n-butyl lithium (2.5 M, 16 mL) was added dropwise, and the mixture was kept warm and stirred for 1 hour. Subsequently, 8.15 g of compound M27-4 was dissolved in 30 mL of tetrahydrofuran and slowly added dropwise to the reaction solution. After the addition, the temperature was raised to -10°C and the reaction was continued for 2.5 hours. The reaction solution was quenched with 200 mL saturated brine, extracted with EtOAc (300 mL×2), and the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 11.06 g of compound M27-5.
步骤3：化合物M27-7的制备Step 3: Preparation of compound M27-7
将11.06g化合物M27-5溶解于水(40mL)和二氯甲烷(200mL)的混合溶液中，随后依次加入3.17g吡啶、12.83g M27-6和0.86g四丁基溴化铵。常温搅拌24小时。反应液用100mL饱和食盐水淬灭，用DCM(100mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，经柱层析纯化得5.46g化合物M27-7。11.06g of compound M27-5 was dissolved in a mixed solution of water (40mL) and dichloromethane (200mL), and then 3.17g of pyridine, 12.83g of M27-6 and 0.86g of tetrabutylammonium bromide were sequentially added. Stir at room temperature for 24 hours. The reaction solution was quenched with 100 mL of saturated brine, extracted with DCM (100 mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 5.46 g of compound M27-7.
步骤4：化合物M27-8的制备Step 4: Preparation of compound M27-8
将5.46g化合物M27-7和2.84g叔丁醇钾溶于85mL无水四氢呋喃中，氮气置换3次后，微波70℃反应5分钟。减压除去溶剂，加入100mL饱和食盐水，再用EtOAc(100mL×2)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，经柱层析纯化得2.91g化合物M27-8。Dissolve 5.46 g of compound M27-7 and 2.84 g of potassium tert-butoxide in 85 mL of anhydrous tetrahydrofuran, replace with nitrogen three times, and react in a microwave at 70° C. for 5 minutes. The solvent was removed under reduced pressure, 100 mL of saturated brine was added, and then extracted with EtOAc (100 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 2.91 g of compound M27-8.
步骤5：化合物M27-9的制备Step 5: Preparation of compound M27-9
将1.30g化合物M27-8，1.56g(R)-(+)-叔丁基亚磺酰胺溶于钛酸四乙酯(10mL)和无水四氢呋喃(2mL)的混合溶液中，氮气置换3次后，90℃反应8小时。停止加热，将反应液倒入200mL冰水中，再加入150mL乙酸乙酯，搅拌1小时。过滤除去滤渣。滤液用乙酸乙酯(100mL×2)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，得1.90g化合物M27-9。Dissolve 1.30g of compound M27-8 and 1.56g of (R)-(+)-tert-butylsulfinamide in a mixed solution of tetraethyl titanate (10mL) and anhydrous tetrahydrofuran (2mL), and replace with nitrogen 3 times Then, it was reacted at 90°C for 8 hours. The heating was stopped, the reaction solution was poured into 200 mL of ice water, 150 mL of ethyl acetate was added, and the mixture was stirred for 1 hour. Filter to remove the filter residue. The filtrate was extracted with ethyl acetate (100 mL×2), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 1.90 g of compound M27-9.
步骤6：化合物M27-10的制备Step 6: Preparation of compound M27-10
将1.30g化合物M27-9溶于25mL无水四氢呋喃中，在-20℃下分批加入344mg硼氢化钠，缓慢升至室温，再常温搅拌3小时。0℃下，加入100mL水，再用EtOAc(80mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，经柱层析纯化得0.94g化合物M27-10。Dissolve 1.30 g of compound M27-9 in 25 mL of anhydrous tetrahydrofuran, add 344 mg of sodium borohydride in batches at -20°C, slowly warm to room temperature, and then stir at room temperature for 3 hours. At 0°C, 100 mL of water was added, and then extracted with EtOAc (80 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by column chromatography to obtain 0.94 g of compound M27-10.
步骤7：化合物M27-11的制备Step 7: Preparation of compound M27-11
将0.94g化合物M27-10溶于10mL二氯甲烷中，再滴加2.62g三氟乙酸，常温搅拌1小时。将反应液降至0℃，再加入饱和碳酸氢钠水溶液调节pH至弱碱性。用EtOAc:THF＝1:1的混合溶液萃取(60mL×4)，合并有机相，无水硫酸钠干燥，减压浓缩。得692mg化合物M27-11。0.94 g of compound M27-10 was dissolved in 10 mL of dichloromethane, and then 2.62 g of trifluoroacetic acid was added dropwise, and the mixture was stirred at room temperature for 1 hour. The reaction solution was reduced to 0°C, and then saturated aqueous sodium bicarbonate solution was added to adjust the pH to weakly alkaline. It was extracted with a mixed solution of EtOAc:THF=1:1 (60 mL×4), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. 692 mg of compound M27-11 was obtained.
实施例19化合物19的制备：Example 19 Preparation of compound 19:
Figure PCTCN2020072774-appb-000042
Figure PCTCN2020072774-appb-000042
步骤1：化合物19-2的制备Step 1: Preparation of compound 19-2
将1.00g化合物19-1溶解于10mL无水乙腈中，加入1.45g碳酸钾，85℃搅拌反应2小时。TLC检测反应完全，减压除去溶剂，加入50mL水，再用EA(50mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得720mg化合物19-2。1.00 g of compound 19-1 was dissolved in 10 mL of anhydrous acetonitrile, 1.45 g of potassium carbonate was added, and the reaction was stirred at 85°C for 2 hours. TLC detected the completion of the reaction, the solvent was removed under reduced pressure, 50 mL of water was added, and then extracted with EA (50 mL×2), the organic layers were combined, dried with sodium sulfate and then desolventized, and purified by column chromatography to obtain 720 mg of compound 19-2.
步骤2：化合物19-3的制备Step 2: Preparation of compound 19-3
将700mg化合物19-2、450mg化合物M1-2、880mg DIEA、63mg Pd 2(dba) 3和100mg Xantphos溶解于10mL二氧六环中，氮气置换3次后，85℃反应12hrs。停止加热，减压浓缩，加入30mL水，再用EA(30mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得271mg化合物19-3。 700 mg of compound 19-2, 450 mg of compound M1-2, 880 mg of DIEA, 63 mg of Pd 2 (dba) 3 and 100 mg of Xantphos were dissolved in 10 mL of dioxane, replaced with nitrogen three times, and reacted at 85°C for 12 hrs. Stop heating, concentrate under reduced pressure, add 30 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 271 mg of compound 19-3.
步骤3：化合物19-4的制备Step 3: Preparation of compound 19-4
氮气保护下，将270mg化合物19-3溶于5mL无水四氢呋喃中，将EtONa的EtOH溶液(2.0mL，20％)缓慢滴加入上述溶液中，剧烈搅拌40min。TLC检测反应完全，将反应液减压浓缩，残余物加入5mL DCM打浆30min，过滤，滤饼用DCM(5mL×3)洗涤，取滤饼真空干燥得154mg棕色固体化合物19-4。Under nitrogen protection, 270 mg of compound 19-3 was dissolved in 5 mL of anhydrous tetrahydrofuran, and EtONa's EtOH solution (2.0 mL, 20%) was slowly added dropwise to the above solution, and stirred vigorously for 40 min. TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was added to 5mL DCM for 30min to be slurried, filtered, the filter cake was washed with DCM (5mL×3), and the filter cake was vacuum-dried to obtain 154 mg of brown solid compound 19-4.
步骤4：化合物19-5的制备Step 4: Preparation of compound 19-5
将44mg化合物19-4、40mg化合物M22-2、4mg Pd 2(dba) 3、5mg Xantphos和21mg DIEA溶于2mL二氧六环中，用氮气置换3次后，加热至100℃反应3小时，TLC跟踪反应。减压除去溶剂，加入30mL水，再用二氯甲烷(30mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得28mg化合物19-5。 Dissolve 44 mg of compound 19-4, 40 mg of compound M22-2, 4 mg of Pd 2 (dba) 3 , 5 mg of Xantphos and 21 mg of DIEA in 2 mL of dioxane, replace it with nitrogen three times, and heat to 100°C for 3 hours. TLC tracks the reaction. The solvent was removed under reduced pressure, 30 mL of water was added, and the mixture was extracted with dichloromethane (30 mL×2). The organic layers were combined, dried over sodium sulfate and desolventized, and purified by column chromatography to obtain 28 mg of compound 19-5.
步骤5：化合物19的制备Step 5: Preparation of compound 19
将28mg化合物19-5，溶于1mL二氧六环中，滴加盐酸甲醇溶液(2N，0.4mL)，TLC跟踪反应。反应完全后，减压除去溶剂，用少量正己烷固化后，倾倒掉正己烷，加入0.5mL水溶解固体，再滴入2滴饱和碳酸氢钠水溶液。过滤得到固体，固体用少量水洗涤后， 真空干燥。得到15.7mg淡黄色固体化合物19。28 mg of compound 19-5 was dissolved in 1 mL of dioxane, and methanol solution of hydrochloric acid (2N, 0.4 mL) was added dropwise, and the reaction was followed by TLC. After the reaction was completed, the solvent was removed under reduced pressure, and after solidification with a small amount of n-hexane, the n-hexane was poured out, 0.5 mL of water was added to dissolve the solid, and 2 drops of saturated sodium bicarbonate aqueous solution were added dropwise. The solid was obtained by filtration, and the solid was washed with a small amount of water and dried in vacuum. 15.7 mg of compound 19 was obtained as a pale yellow solid.
[M+H +]＝467.29 [M+H + ]=467.29
1H NMR(500MHz,DMSO-d 6,):δ8.01(s,1H),7.31(d,J＝6.6Hz,1H),7.22-7.08(m,5H),6.91(t,J＝7.0Hz,1H),5.35(t,J＝5.0Hz,1H),4.48(d,J＝5.2Hz,2H),3.85(s,3H),3.68-3.64(m,2H),3.33-3.30(m,1H),3.10-3.00(m,3H),2.64-2.59(m,1H),1.91-1.86(m,1H),1.80-1.76(m,1H),1.51(d,J＝12.8Hz,1H),1.13-1.10(m,1H). 1 H NMR(500MHz,DMSO-d 6 ,):δ8.01(s,1H),7.31(d,J=6.6Hz,1H),7.22-7.08(m,5H),6.91(t,J=7.0 Hz, 1H), 5.35 (t, J = 5.0 Hz, 1H), 4.48 (d, J = 5.2 Hz, 2H), 3.85 (s, 3H), 3.68-3.64 (m, 2H), 3.33-3.30 (m ,1H),3.10-3.00(m,3H),2.64-2.59(m,1H),1.91-1.86(m,1H),1.80-1.76(m,1H),1.51(d,J=12.8Hz,1H ), 1.13-1.10 (m, 1H).
化合物20的制备：Preparation of compound 20:
Figure PCTCN2020072774-appb-000043
Figure PCTCN2020072774-appb-000043
步骤1：化合物20-2的制备Step 1: Preparation of compound 20-2
将500mg化合物20-1和318mg化合物M1-2溶解于6mL二氧六环中，加入48mg Pd 2(dba) 3、76mg Xantphos和684mg DIPEA，氮气置换三次，反应液升至90℃搅拌反应5hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得569mg化合物20-2。 500 mg of compound 20-1 and 318 mg of compound M1-2 were dissolved in 6 mL of dioxane, 48 mg of Pd 2 (dba) 3 , 76 mg of Xantphos, and 684 mg of DIPEA were added, and nitrogen was replaced three times. The reaction solution was heated to 90° C. and stirred for 5 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 569 mg of compound 20-2.
步骤2：化合物20-3的制备Step 2: Preparation of compound 20-3
将569mg化合物20-2溶解于7mL THF中，-30℃下滴加EtONa(1.3mL，20％的EtOH溶液)，RT搅拌反应3hrs。TLC检测反应完全，减压浓缩，加入10mL DCM搅拌30mins，反应液过滤，滤饼用DCM(10mL×2)洗涤，抽干得380mg化合物20-3。569 mg of compound 20-2 was dissolved in 7 mL of THF, EtONa (1.3 mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 3 hrs. TLC detected the completion of the reaction, concentrated under reduced pressure, added 10 mL DCM and stirred for 30 mins, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), and drained to obtain 380 mg of compound 20-3.
步骤3：化合物20-4的制备Step 3: Preparation of compound 20-4
将35mg化合物20-3和70mg化合物M22-2溶解于1mL二氧六环中，加入7mg Pd 2(dba) 3、8mg Xantphos和37mg DIPEA。氮气置换三次，氮气保护下反应升至90℃搅 拌反应12hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得55mg化合物20-4。 35 mg of compound 20-3 and 70 mg of compound M22-2 were dissolved in 1 mL of dioxane, and 7 mg of Pd 2 (dba) 3 , 8 mg of Xantphos and 37 mg of DIPEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 55 mg of compound 20-4.
步骤4：化合物20的制备Step 4: Preparation of compound 20
将55mg化合物20-4溶解于1mL二氧六环和0.3mL MeOH中，加入2N HCl(0.25mL，甲醇溶液)，RT搅拌反应2hrs。TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(1.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(1.5mL)洗涤，取滤饼真空干燥得12mg淡黄色固体化合物20。 55 mg of compound 20-4 was dissolved in 1 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.25 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H 2 O (1.5 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (1.5 mL), take the filter cake and vacuum dry to obtain 12 mg of light yellow solid Compound 20.
[M+H +]＝451.24。 [M+H + ]=451.24.
1H NMR(500MHz,DMSO-d6)δ8.36(s,1H),8.32(d,J＝5.3Hz,1H),7.38-7.33(m,1H),7.24-7.17(m,3H),6.82(d,J＝5.3Hz,1H),5.50(d,J＝6.3Hz,1H),4.53(d,J＝4.6Hz,2H),3.98(s,1H),3.93-3.86(m,2H),3.80(s,3H),3.24-3.16(m,2H),3.08(d,J＝15.7Hz,1H),2.71(d,J＝15.7Hz,1H),1.92-1.85(m,1H),1.84-1.77(m,1H),1.58-1.52(m,1H),1.31-1.27(m,1H). 1 H NMR(500MHz,DMSO-d6)δ8.36(s,1H),8.32(d,J=5.3Hz,1H),7.38-7.33(m,1H),7.24-7.17(m,3H),6.82 (d,J=5.3Hz,1H), 5.50(d,J=6.3Hz,1H), 4.53(d,J=4.6Hz,2H), 3.98(s,1H),3.93-3.86(m,2H) ,3.80(s,3H),3.24-3.16(m,2H),3.08(d,J=15.7Hz,1H), 2.71(d,J=15.7Hz,1H),1.92-1.85(m,1H), 1.84-1.77 (m, 1H), 1.58-1.52 (m, 1H), 1.31-1.27 (m, 1H).
实施例21化合物21的制备：Example 21 Preparation of compound 21:
Figure PCTCN2020072774-appb-000044
Figure PCTCN2020072774-appb-000044
步骤1：化合物21-1A的制备Step 1: Preparation of compound 21-1A
将200mg化合物M22-SM和249mg化合物M6溶解于2.5mL THF中，加入137mg三乙胺。RT搅拌反应4hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得222mg化合物21-1A。200 mg of compound M22-SM and 249 mg of compound M6 were dissolved in 2.5 mL of THF, and 137 mg of triethylamine was added. The reaction was stirred at RT for 4hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 222 mg of compound 21-1A.
步骤2：化合物21-2A的制备Step 2: Preparation of compound 21-2A
将222mg化合物21-1A和109mg化合物M1溶解于5mL二氧六环中，加入19mgPd 2(dba) 3，25mg Xantphos。氮气置换三次，氮气保护下反应升至80℃搅拌反应12hrs。TLC 检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得145mg化合物21-2A。 222 mg of compound 21-1A and 109 mg of compound M1 were dissolved in 5 mL of dioxane, 19 mg of Pd 2 (dba) 3 and 25 mg of Xantphos were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 80°C and stirred for 12 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 145 mg of compound 21-2A.
步骤3：化合物21-3A的制备Step 3: Preparation of compound 21-3A
将60mg化合物21-2A溶解于1mL的无水THF中。在0℃下，分批加入11mg LiAlH 4，0℃搅拌反应0.5hrs。TLC检测反应完全，将反应液倒入10mL冰水中，加DCM萃取(15mL×3)，合并有机相，用15mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经TLC板纯化得12mg白色固体化合物21-3A。 60 mg of compound 21-2A was dissolved in 1 mL of anhydrous THF. At 0°C, 11 mg LiAlH 4 was added in batches, and the reaction was stirred at 0°C for 0.5 hrs. TLC detects the completion of the reaction. Pour the reaction solution into 10 mL ice water, add DCM to extract (15 mL×3), combine the organic phases, wash with 15 mL saturated NaCl, dry with anhydrous sodium sulfate, and concentrate under reduced pressure. The residue is purified by TLC plate. 12 mg of white solid compound 21-3A.
步骤4：化合物21的制备Step 4: Preparation of compound 21
将12mg化合物21-3A溶解于0.5mL二氧六环和0.1mL MeOH中，加入2N HCl(0.1mL，甲醇溶液)，RT搅拌反应2hrs。TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(0.5mL)洗涤，取滤饼真空干燥得7mg白色固体化合物21。 12 mg of compound 21-3A was dissolved in 0.5 mL of dioxane and 0.1 mL of MeOH, 2N HCl (0.1 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete and the reaction solution was concentrated under reduced pressure. Add H 2 O (0.5 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (0.5 mL), take the filter cake and vacuum dry to obtain 7 mg of white solid compound twenty one.
[M+H +]＝469.26。 [M+H + ]=469.26.
1H NMR(500MHz,DMSO-d 6)δ8.31(s,1H),7.67(d,J＝5.4Hz,1H),7.31(d,J＝6.8Hz,1H),7.23-7.11(m,3H),6.36(s,2H),5.91(d,J＝5.3Hz,1H),5.48(t,J＝5.9Hz,1H),4.52(d,J＝5.9Hz,2H),3.94-3.83(m,3H),3.24-3.11(m,2H),3.06(d,J＝15.5Hz,1H),2.63(d,J＝15.3Hz,1H),1.94-1.85(m,1H),1.83-1.75(m,1H),1.59-1.50(m,1H),1.18-1.07(m,1H). 1 H NMR(500MHz,DMSO-d 6 )δ8.31(s,1H), 7.67(d,J=5.4Hz,1H), 7.31(d,J=6.8Hz,1H), 7.23-7.11(m, 3H), 6.36(s, 2H), 5.91(d,J=5.3Hz,1H), 5.48(t,J=5.9Hz,1H), 4.52(d,J=5.9Hz,2H),3.94-3.83( m,3H),3.24-3.11(m,2H),3.06(d,J=15.5Hz,1H), 2.63(d,J=15.3Hz,1H),1.94-1.85(m,1H),1.83-1.75 (m, 1H), 1.59-1.50 (m, 1H), 1.18-1.07 (m, 1H).
实施例22化合物22的制备：Example 22 Preparation of Compound 22:
Figure PCTCN2020072774-appb-000045
Figure PCTCN2020072774-appb-000045
步骤1：化合物22-2的制备Step 1: Preparation of compound 22-2
氮气氛围下，将600mg化合物22-1和414mg化合物M1-2溶解于10mL二氧六环中，加入105mg Pd 2(dba) 3、166mg Xantphos和1.49g DIEA,升温至85℃反应3hrs。TLC检测反应完全，浓缩，加入50mL乙酸乙酯和50mL水分散，水相加50mL乙酸乙酯萃取，合 并有机相，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化，得类白色固体0.605g化合物22-2。 Under a nitrogen atmosphere, 600 mg of compound 22-1 and 414 mg of compound M1-2 were dissolved in 10 mL of dioxane, 105 mg of Pd 2 (dba) 3 , 166 mg of Xantphos and 1.49 g of DIEA were added, and the temperature was raised to 85° C. to react for 3 hrs. TLC detects that the reaction is complete, concentrate, add 50mL ethyl acetate and 50mL water to disperse, add 50mL ethyl acetate to extract the aqueous phase, combine the organic phases, dry with anhydrous sodium sulfate, concentrate under reduced pressure, the residue is purified by column chromatography to obtain a class 0.605 g of compound 22-2 as a white solid.
步骤2：化合物22-3的制备Step 2: Preparation of compound 22-3
将0.605g化合物22-2溶解于5mL THF中，RT滴加EtONa(5.75mL，20％的EtOH溶液)，RT搅拌反应3hrs。TLC检测反应完全，减压浓缩，加入15mL的正己烷搅拌30mins，反应液过滤，滤饼用正己烷(15mL×2)洗涤，得米黄色固体660mg化合物22-3。0.605g compound 22-2 was dissolved in 5mL THF, EtONa (5.75mL, 20% EtOH solution) was added dropwise at RT, and the reaction was stirred at RT for 3hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, added 15 mL of n-hexane and stirred for 30 mins, the reaction solution was filtered, and the filter cake was washed with n-hexane (15 mL×2) to obtain 660 mg of compound 22-3 as a beige solid.
步骤3：化合物22-4的制备Step 3: Preparation of compound 22-4
氮气氛围下，将50mg化合物M22-2和22mg化合物22-3溶解于10mL二氧六环中，加入9.28mg Pd 2(dba) 3、11.73mg Xantphos和26mg DIEA,升温至100℃反应3hrs。TLC检测反应完全，浓缩，残余物经制备薄层色谱法纯化得8.4mg白色化合物22-4。 Under a nitrogen atmosphere, 50 mg of compound M22-2 and 22 mg of compound 22-3 were dissolved in 10 mL of dioxane, 9.28 mg of Pd 2 (dba) 3 , 11.73 mg of Xantphos and 26 mg of DIEA were added, and the temperature was raised to 100° C. to react for 3 hours. TLC detected that the reaction was complete, concentrated, and the residue was purified by preparative thin-layer chromatography to obtain 8.4 mg of white compound 22-4.
步骤3：化合物22的制备Step 3: Preparation of compound 22
氮气氛围下，将8.4mg化合物22-5溶解于1.5mL二氧六环中和0.5mL MeOH中，加入2N HCl(0.32mL，甲醇溶液)，RT搅拌反应1hrs，TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(2mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(2mL)洗涤，取滤饼真空干燥得1.2mg米黄色固体化合物22。 Under a nitrogen atmosphere, 8.4 mg of compound 22-5 was dissolved in 1.5 mL of dioxane and 0.5 mL of MeOH, and 2N HCl (0.32 mL, methanol solution) was added. The reaction was stirred at RT for 1 hrs. TLC detected that the reaction was complete and the reaction solution was reduced. Pressure concentration. Add H 2 O (2 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (2 mL), take the filter cake and vacuum dry to obtain 1.2 mg of beige solid compound twenty two.
[M+H +]＝436.32。 [M+H + ]=436.32.
1H NMR(500MHz,DMSO-d 6)δ8.33(s,1H),7.96(d,J＝5.0Hz,1H),7.31(d,J＝7.0Hz,1H),7.19-7.14(m,3H),6.70(s,2H),6.09(d,J＝5.0Hz,1H),5.47(t,J＝5.0Hz,1H),4.52(d,J＝5.5Hz,2H),3.89-3.82(m,3H),3.21-3.11(m,2H),3.06(d,J＝16.0Hz,1H),2.63(d,J＝15.5Hz,1H),1.93-1.87(m,1H),1.83-1.76(m,1H),1.56-1.53(m,1H),1.16-1.13(m,1H). 1 H NMR(500MHz,DMSO-d 6 )δ8.33(s,1H),7.96(d,J=5.0Hz,1H), 7.31(d,J=7.0Hz,1H), 7.19-7.14(m, 3H), 6.70 (s, 2H), 6.09 (d, J = 5.0 Hz, 1H), 5.47 (t, J = 5.0 Hz, 1H), 4.52 (d, J = 5.5 Hz, 2H), 3.89-3.82 ( m,3H),3.21-3.11(m,2H),3.06(d,J=16.0Hz,1H), 2.63(d,J=15.5Hz,1H),1.93-1.87(m,1H),1.83-1.76 (m,1H),1.56-1.53(m,1H),1.16-1.13(m,1H).
实施例23化合物23的制备：Example 23 Preparation of compound 23:
Figure PCTCN2020072774-appb-000046
Figure PCTCN2020072774-appb-000046
步骤1：化合物21-A1的制备Step 1: Preparation of compound 21-A1
将1.61g化合物M22-SM和2.00g化合物M6溶解于20mL THF中，加入1.10g三乙胺，RT搅拌反应12hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(30mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得2.00g化合物21-A1。1.61 g of compound M22-SM and 2.00 g of compound M6 were dissolved in 20 mL of THF, 1.10 g of triethylamine was added, and the reaction was stirred at RT for 12 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (30 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 2.00 g of compound 21-A1.
步骤2：化合物23-1的制备Step 2: Preparation of compound 23-1
将100mg化合物21-1A溶解于1mL THF中，滴加24mg LiOH·H 2O的H 2O(1mL)溶液，RT搅拌反应12hrs。TLC检测反应完全，用1N HCl溶液调节反应液至pH＝7，加DCM萃取(10mL×3)，合并有机相，用15mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得94mg化合物23-1。 100 mg of compound 21-1A was dissolved in 1 mL of THF, 24 mg of LiOH·H 2 O in H 2 O (1 mL) was added dropwise, and the reaction was stirred at RT for 12 hrs. TLC detects the completion of the reaction. Adjust the reaction solution to pH=7 with 1N HCl solution, add DCM to extract (10mL×3), combine the organic phases, wash with 15mL saturated NaCl, dry with anhydrous sodium sulfate, concentrate under reduced pressure, and pass the residue through a column Chromatographic purification yielded 94 mg of compound 23-1.
步骤3：化合物23-3的制备Step 3: Preparation of compound 23-3
将94mg化合物23-1溶解于1.5mL的无水DMF中。在0℃下，加入46mg EDCI、33mg HOBT、24mg TEA和17mg化合物23-2。氮气保护下RT搅拌反应5hrs。TLC检测反应完全，将反应液倒入5mL水中，加EtOAc萃取(15mL×2)，合并有机相，用15mL饱和NaCl洗涤三次，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得65mg化合物23-3。94 mg of compound 23-1 was dissolved in 1.5 mL of anhydrous DMF. At 0°C, add 46mg EDCI, 33mg HOBT, 24mg TEA and 17mg compound 23-2. The reaction was stirred at RT for 5hrs under nitrogen protection. The reaction was completed by TLC detection, the reaction solution was poured into 5mL water, extracted with EtOAc (15mL×2), the organic phases were combined, washed with 15mL saturated NaCl three times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography 65 mg of compound 23-3 was obtained.
步骤4：化合物23-4的制备Step 4: Preparation of compound 23-4
将65mg化合物23-3溶解于1.8mL THF中，-78℃下滴加MgBrCH 3(3M，0.196mL)反应液升至0℃搅拌反应1hrs。TLC检测反应完全，用饱和NH 4Cl(8mL)淬灭反应，加EtOAc萃取(15mL×2)，合并有机相，用15mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经TLC板纯化得10mg化合物23-4。 65 mg of compound 23-3 was dissolved in 1.8 mL of THF, and MgBrCH 3 (3M, 0.196 mL) was added dropwise at -78° C. The reaction solution was raised to 0° C. and stirred for 1 hrs. The reaction was completed by TLC detection. The reaction was quenched with saturated NH 4 Cl (8 mL), extracted with EtOAc (15 mL×2), and the organic phases were combined, washed with 15 mL saturated NaCl, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was subjected to TLC Plate purification yielded 10 mg of compound 23-4.
步骤5：化合物23-5的制备Step 5: Preparation of compound 23-5
将10mg化合物23-4和6mg化合物M1溶解于0.5mL二氧六环中，加入1mg Pd 2(dba) 3，1mg Xantphos和5mg DIPEA。氮气置换三次，氮气保护下反应升至90℃搅拌反应12hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(5mL×2)洗涤，滤液减压浓缩，残余物经TLC板纯化得8mg化合物23-5。 10 mg of compound 23-4 and 6 mg of compound M1 were dissolved in 0.5 mL of dioxane, and 1 mg Pd 2 (dba) 3 , 1 mg Xantphos and 5 mg DIPEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 90°C and stirred for 12hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (5 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by TLC plate to obtain 8 mg of compound 23-5.
步骤6：化合物23的制备Step 6: Preparation of compound 23
将8mg化合物23-5溶解于0.4mL二氧六环和0.1mL MeOH中，加入2N HCl(0.1mL，甲醇溶液)，RT搅拌反应2hrs。TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(0.5mL)洗涤，取滤饼真空干燥得2.1mg黄色固体化合物23。 8 mg of compound 23-5 was dissolved in 0.4 mL of dioxane and 0.1 mL of MeOH, 2N HCl (0.1 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H 2 O (0.5 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (0.5 mL), take the filter cake and vacuum dry to obtain 2.1 mg of yellow solid Compound 23.
[M+H +]＝481.23。 [M+H + ]=481.23.
1H NMR(500MHz,DMSO-d 6)δ8.41(s,1H),7.70(d,J＝5.3Hz,1H),7.30(d,J＝6.8Hz,1H),7.22-7.13(m,3H),6.37(s,2H),5.97(d,J＝5.4Hz,1H),3.90-3.78(m,3H),3.24(q,J＝13.5,13.0Hz,2H),3.07(d,J＝15.4Hz,1H),2.65(d,J＝15.3Hz,1H),2.56(s,3H),1.85-1.78(m,1H),1.76-1.69(m,1H),1.56-1.50(m,1H),1.18-1.11(m,1H). 1 H NMR(500MHz,DMSO-d 6 )δ8.41(s,1H), 7.70(d,J=5.3Hz,1H), 7.30(d,J=6.8Hz,1H), 7.22-7.13(m, 3H), 6.37 (s, 2H), 5.97 (d, J = 5.4 Hz, 1H), 3.90-3.78 (m, 3H), 3.24 (q, J = 13.5, 13.0 Hz, 2H), 3.07 (d, J = 15.4Hz, 1H), 2.65 (d, J = 15.3Hz, 1H), 2.56 (s, 3H), 1.85-1.78 (m, 1H), 1.76-1.69 (m, 1H), 1.56-1.50 (m, 1H), 1.18-1.11 (m, 1H).
实施例24：化合物24的制备：Example 24: Preparation of Compound 24:
Figure PCTCN2020072774-appb-000047
Figure PCTCN2020072774-appb-000047
步骤1：化合物24-6的制备Step 1: Preparation of compound 24-6
将90mg化合物M16-5和100mg化合物M22-2溶解于1.5mL二氧六环中，加入5mg Pd 2(dba) 3，6mg Xantphos和26mg DIPEA。氮气置换三次，氮气保护下反应升至100℃搅拌反应5hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得84mg化合物24-6。 90 mg of compound M16-5 and 100 mg of compound M22-2 were dissolved in 1.5 mL of dioxane, and 5 mg Pd 2 (dba) 3 , 6 mg Xantphos and 26 mg DIPEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 100°C and stirred for 5 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 84 mg of compound 24-6.
步骤2：化合物24的制备Step 2: Preparation of compound 24
将84mg化合物24-6溶解于1.5mL二氧六环和0.5mL MeOH中，加入2N HCl(0.32mL，甲醇溶液)，RT搅拌反应2hrs。TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(2mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(2mL)洗涤，取滤饼真空干燥得55mg白色固体24。 84 mg compound 24-6 was dissolved in 1.5 mL dioxane and 0.5 mL MeOH, 2N HCl (0.32 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. The residue was dissolved by adding H 2 O (2 mL), and the solution was adjusted to pH=8 with saturated NaHCO 3. The solid precipitated out and filtered. The filter cake was washed with H 2 O (2 mL), and the filter cake was vacuum-dried to obtain 55 mg of white solid 24.
[M+H +]＝549.22。 [M+H + ]=549.22.
1H NMR(500MHz,DMSO-d 6)δ8.51(s,1H),8.35(s,1H),7.76(d,J＝5.4Hz,1H),7.36(d,J＝2.0Hz,2H),7.25-7.16(m,3H),6.16(d,J＝5.3Hz,1H),6.09(d,J＝1.9Hz,1H),5.49(t,J＝5.9Hz,1H),4.54(d,J＝5.7Hz,2H),3.97(s,1H),3.95-3.87(m,2H),3.56(s,3H),3.23-3.16(m,2H),3.08(d,J＝15.7Hz,1H),2.71(d,J＝15.7Hz,1H),1.93-1.85(m,1H),1.85-1.78(m,1H),1.58-1.52(m,1H),1.24-1.23(m,1H). 1 H NMR(500MHz,DMSO-d 6 )δ8.51(s,1H),8.35(s,1H),7.76(d,J=5.4Hz,1H),7.36(d,J=2.0Hz,2H) ,7.25-7.16(m,3H),6.16(d,J=5.3Hz,1H), 6.09(d,J=1.9Hz,1H), 5.49(t,J=5.9Hz,1H),4.54(d, J=5.7Hz,2H),3.97(s,1H),3.95-3.87(m,2H),3.56(s,3H),3.23-3.16(m,2H),3.08(d,J=15.7Hz,1H ), 2.71(d,J=15.7Hz,1H),1.93-1.85(m,1H),1.85-1.78(m,1H),1.58-1.52(m,1H),1.24-1.23(m,1H).
实施例25化合物25的制备：Example 25 Preparation of compound 25:
Figure PCTCN2020072774-appb-000048
Figure PCTCN2020072774-appb-000048
步骤1：化合物25-2的制备Step 1: Preparation of compound 25-2
将500mg化合物M20-SM和186mg化合物25-1溶解于5mL DMSO中，氮气置换三次，氮气保护下反应升至70℃搅拌反应4hrs。TLC检测反应完全，将反应液倒入20mL水中，加EtOAc萃取(30mL×2)，合并有机相，用30mL饱和NaCl洗涤三次，无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得490mg白色固体物化合物25-2。500 mg of compound M20-SM and 186 mg of compound 25-1 were dissolved in 5 mL of DMSO, replaced with nitrogen three times, and the reaction was raised to 70°C under nitrogen protection and stirred for 4 hrs. TLC detected the completion of the reaction. The reaction solution was poured into 20mL water, extracted with EtOAc (30mL×2), combined the organic phases, washed with 30mL saturated NaCl three times, dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by column chromatography 490 mg of white solid compound 25-2 was obtained.
步骤2：化合物25-3的制备Step 2: Preparation of compound 25-3
将490mg化合物25-2和181.43mg化合物M1-2溶解于6mL二氧六环中，加入28mg Pd 2(dba) 3、44mg Xantphos和390mg DIPEA，氮气置换三次，反应液升至90℃搅拌反应12hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得478mg化合物25-3。 Dissolve 490 mg of compound 25-2 and 181.43 mg of compound M1-2 in 6 mL of dioxane, add 28 mg of Pd 2 (dba) 3 , 44 mg of Xantphos and 390 mg of DIPEA, replace with nitrogen three times, and raise the reaction solution to 90°C and stir for 12 hours. . TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 478 mg of compound 25-3.
步骤3：化合物25-4的制备Step 3: Preparation of compound 25-4
将478mg化合物25-3溶解于5mL THF中，-30℃下滴加EtONa(2mL，20％的EtOH溶液)，RT搅拌反应4hrs。TLC检测反应完全，减压浓缩，加入20mL DCM搅拌30mins，反应液过滤，滤饼用DCM(10mL×2)洗涤，抽干得449mg化合物25-4。478mg of compound 25-3 was dissolved in 5mL THF, EtONa (2mL, 20% EtOH solution) was added dropwise at -30°C, and the reaction was stirred at RT for 4hrs. TLC detected the reaction to be complete, concentrated under reduced pressure, added 20mL DCM and stirred for 30mins, the reaction solution was filtered, the filter cake was washed with DCM (10mL×2), and drained to obtain 449mg of compound 25-4.
步骤4：化合物25-5的制备Step 4: Preparation of compound 25-5
将32mg化合物25-4和50mg化合物M22-2溶解于1mL二氧六环中，加入5mg Pd 2(dba) 3，6mg Xantphos和26mg DIPEA。氮气置换三次，氮气保护下反应升至100℃搅拌反应3hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得25mg化合物25-5。 32 mg of compound 25-4 and 50 mg of compound M22-2 were dissolved in 1 mL of dioxane, and 5 mg of Pd 2 (dba) 3 , 6 mg of Xantphos and 26 mg of DIPEA were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 100°C and stirred for 3hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 25 mg of compound 25-5.
步骤5：化合物25的制备Step 5: Preparation of compound 25
将25mg化合物25-5溶解于0.7mL二氧六环和0.2mL MeOH中，加入2N HCl(0.1mL，甲醇溶液)，RT搅拌反应2hrs。TLC检测反应完全，反应液减压浓缩。残余物加入H 2O (1mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(1mL)洗涤，取滤饼真空干燥得15mg白色固体化合物25。 25 mg of compound 25-5 was dissolved in 0.7 mL of dioxane and 0.2 mL of MeOH, 2N HCl (0.1 mL, methanol solution) was added, and the reaction was stirred at RT for 2 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. The residue was dissolved by adding H 2 O (1 mL), the solution was adjusted to pH=8 with saturated NaHCO 3 , the solid was precipitated, filtered, the filter cake was washed with H 2 O (1 mL), and the filter cake was vacuum dried to obtain 15 mg of white solid compound 25.
[M+H +]＝539.34。 [M+H + ]=539.34.
1H NMR(500MHz,DMSO-d 6)δ8.34(s,1H),7.99(d,J＝5.3Hz,1H),7.31(d,J＝6.9Hz,1H),7.20-7.12(m,3H),6.40(d,J＝5.4Hz,1H),5.47(t,J＝5.9Hz,1H),4.53(d,J＝5.8Hz,2H),3.93-3.88(m,3H),3.75-3.72(m,4H),3.23-3.12(m,6H),3.06(d,J＝15.6Hz,1H),2.62(d,J＝15.3Hz,1H),1.93-1.86(m,1H),1.83-1.76(m,1H),1.58-1.52(m,1H),1.17-1.11(m,1H). 1 H NMR(500MHz,DMSO-d 6 )δ8.34(s,1H),7.99(d,J=5.3Hz,1H), 7.31(d,J=6.9Hz,1H), 7.20-7.12(m, 3H), 6.40(d,J=5.4Hz,1H), 5.47(t,J=5.9Hz,1H), 4.53(d,J=5.8Hz,2H),3.93-3.88(m,3H),3.75- 3.72(m,4H),3.23-3.12(m,6H),3.06(d,J=15.6Hz,1H), 2.62(d,J=15.3Hz,1H),1.93-1.86(m,1H),1.83 -1.76 (m, 1H), 1.58-1.52 (m, 1H), 1.17-1.11 (m, 1H).
实施例26化合物26的制备：Example 26 Preparation of compound 26:
Figure PCTCN2020072774-appb-000049
Figure PCTCN2020072774-appb-000049
步骤1：化合物26-2的制备Step 1: Preparation of compound 26-2
将0.50g化合物M14和0.39g化合物M22-SM溶解于10mL四氢呋喃中，滴加0.26g DIEA，60℃搅拌反应12小时。TLC检测反应完全，减压浓缩，加入20mL水，再用EtOAc(20mL×3)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得0.58g化合物26-2。0.50g compound M14 and 0.39g compound M22-SM were dissolved in 10mL tetrahydrofuran, 0.26g DIEA was added dropwise, and the reaction was stirred at 60°C for 12 hours. TLC detected the completion of the reaction, concentrated under reduced pressure, added 20 mL of water, and extracted with EtOAc (20 mL×3). The organic layers were combined, dried over sodium sulfate and desolventized, and purified by column chromatography to obtain 0.58 g of compound 26-2.
步骤2：化合物26-3的制备Step 2: Preparation of compound 26-3
氮气保护下，将0.30g化合物26-2溶解于10mL无水二氯甲烷中，温度降至-78℃，滴加DIBAL-H(1M,2.6mL)的正己烷溶液。-78℃搅拌反应1小时。再缓慢升温至-40℃，继续反应2小时。TLC检测反应完全，在0℃下，缓慢滴加0.1mL水，再滴加氢氧化钠水溶液(15％,0.1mL)，再加入0.25mL水。升至室温搅拌15分钟。加入无水硫酸钠干燥，搅拌十分钟后过滤。滤液浓缩，经柱层析纯化得0.21g化合物26-3。Under the protection of nitrogen, 0.30 g of compound 26-2 was dissolved in 10 mL of anhydrous dichloromethane, the temperature was reduced to -78° C., and DIBAL-H (1M, 2.6 mL) in n-hexane was added dropwise. The reaction was stirred at -78°C for 1 hour. Then slowly increase the temperature to -40°C and continue the reaction for 2 hours. TLC detected that the reaction was complete. At 0°C, 0.1 mL of water was slowly added dropwise, then sodium hydroxide aqueous solution (15%, 0.1 mL) was added dropwise, and then 0.25 mL of water was added. Warm to room temperature and stir for 15 minutes. Add anhydrous sodium sulfate to dry, stir for ten minutes and filter. The filtrate was concentrated and purified by column chromatography to obtain 0.21 g of compound 26-3.
步骤3：化合物26-4的制备Step 3: Preparation of compound 26-4
将80mg化合物26-3、52mg化合物M-1溶解于4mL的二氧六环中，加入13mg Pd 2(dba) 3、16mg Xantphos和130mg DIEA，然后100℃反应20hrs。LCMS检测反应完全，将反应液过滤，减压浓缩，残余物用制备板纯化得35mg黄色固体化合物26-4。 80 mg of compound 26-3 and 52 mg of compound M-1 were dissolved in 4 mL of dioxane, 13 mg Pd 2 (dba) 3 , 16 mg Xantphos and 130 mg DIEA were added, and then reacted at 100° C. for 20 hrs. LCMS detected that the reaction was complete, the reaction solution was filtered, concentrated under reduced pressure, and the residue was purified with a preparation plate to obtain 35 mg of yellow solid compound 26-4.
步骤4：化合物26的制备Step 4: Preparation of compound 26
将35mg化合物26-4，溶于2mL二氧六环中，滴加盐酸甲醇溶液(2N,0.2mL)，TLC跟踪反应。反应完全后，减压除去溶剂，用少量正己烷固化后，倾倒掉正己烷，加入1.0mL水溶解固体，再滴入饱和碳酸氢钠水溶液调pH＝9-10。过滤得到固体，固体用少量水洗涤后，真空干燥，得到20.8mg淡黄色固体化合物26。35 mg of compound 26-4 was dissolved in 2 mL of dioxane, and methanol solution of hydrochloric acid (2N, 0.2 mL) was added dropwise, and the reaction was followed by TLC. After the reaction was completed, the solvent was removed under reduced pressure, and after solidification with a small amount of n-hexane, the n-hexane was poured out, 1.0 mL of water was added to dissolve the solid, and then saturated sodium bicarbonate aqueous solution was added dropwise to adjust pH=9-10. The solid was filtered, washed with a small amount of water, and dried in vacuo to obtain 20.8 mg of compound 26 as a pale yellow solid.
[M+H] +:537.16 [M+H] + :537.16
1H NMR(500MHz,DMSO-d 6)δ8.32(s,1H),7.70–7.64(m,2H),7.53(d,J＝7.8Hz,1H),7.42(d,J＝7.8Hz,1H),6.35(s,2H),5.92(d,J＝5.4Hz,1H),5.49(t,J＝5.9Hz,1H),4.53(d,J＝5.8Hz,2H),4.01-3.84(m,3H),3.23-3.13(m,3H),2.75-2.72(m,1H),1.97-1.90(m,1H),1.83-1.80(m,1H),1.60-1.57(m,1H),1.13-1.10(m,1H). 1 H NMR(500MHz,DMSO-d 6 )δ8.32(s,1H), 7.70–7.64(m,2H), 7.53(d,J=7.8Hz,1H), 7.42(d,J=7.8Hz, 1H), 6.35 (s, 2H), 5.92 (d, J = 5.4 Hz, 1H), 5.49 (t, J = 5.9 Hz, 1H), 4.53 (d, J = 5.8 Hz, 2H), 4.01-3.84 ( m,3H),3.23-3.13(m,3H),2.75-2.72(m,1H),1.97-1.90(m,1H),1.83-1.80(m,1H),1.60-1.57(m,1H), 1.13-1.10 (m, 1H).
实施例27化合物27的制备：Example 27 Preparation of compound 27:
Figure PCTCN2020072774-appb-000050
Figure PCTCN2020072774-appb-000050
步骤1：化合物27-1的制备Step 1: Preparation of compound 27-1
将400mg化合物M23-1溶解于20.0mL THF中，冰浴下冷却至0℃，将甲基溴化镁(2.0M，2ml)缓慢加入上述溶液中，升至室温搅拌反应0.5hrs。TLC检测反应完全，将饱和氯化铵水溶液(5mL)缓慢加入反应液中淬灭反应，加入EA(10mL*3)萃取，合并有机相，饱和食盐水(15mL)洗涤，无水硫酸钠干燥，过滤，滤液旋干，残余物经柱层析纯化得60mg白色固体化合物27-1。400mg of compound M23-1 was dissolved in 20.0mL THF, cooled to 0°C in an ice bath, methylmagnesium bromide (2.0M, 2ml) was slowly added to the above solution, and the temperature was raised to room temperature and stirred for 0.5hrs. TLC detected that the reaction was complete. Saturated aqueous ammonium chloride solution (5 mL) was slowly added to the reaction solution to quench the reaction, EA (10 mL*3) was added for extraction, the organic phases were combined, washed with saturated brine (15 mL), and dried with anhydrous sodium sulfate. After filtration, the filtrate was spin-dried, and the residue was purified by column chromatography to obtain 60 mg of white solid compound 27-1.
步骤2：化合物27-2的制备Step 2: Preparation of compound 27-2
将60mg化合物27-1和44mg化合物M-1溶解于5mL二氧六环中，加入11mg Pd 2(dba) 3、14mg Xantphos、46mg N,N-二异丙基乙胺。氮气置换三次，氮气保护下反应升至100℃搅拌反应18hrs。LCMS和TLC检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得21mg化合物27-2。 60 mg of compound 27-1 and 44 mg of compound M-1 were dissolved in 5 mL of dioxane, 11 mg of Pd 2 (dba) 3 , 14 mg of Xantphos, and 46 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 100°C and stirred for 18 hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 21 mg of compound 27-2.
步骤3：化合物27的制备Step 3: Preparation of compound 27
将21mg化合物27-2溶解于1.5mL二氧六环和0.3mL MeOH中，加入2N HCl(0.5mL，甲醇溶液)，RT搅拌反应0.5hrs。LCMS和TLC检测反应完全，反应液减压浓缩。残余物加入 H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8-9，固体析出，过滤，滤饼用H 2O(0.5mL)洗涤，固体再用DCM溶解，干燥后真空浓缩，经制备板纯化得16.3mg白色固体27。 21 mg of compound 27-2 was dissolved in 1.5 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.5 mL, methanol solution) was added, and the reaction was stirred at RT for 0.5 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H 2 O (0.5 mL) to the residue to dissolve it, adjust the solution to pH=8-9 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (0.5 mL), dissolve the solid with DCM, and dry it It was concentrated in vacuo and purified by a preparation plate to obtain 16.3 mg of white solid 27.
[M+H +]＝483.21。 [M+H + ]=483.21.
1H NMR(500MHz,DMSO-d 6)δ8.32(s,1H),7.67(d,J＝5.5Hz,1H),7.31(d,J＝7.0Hz,1H),7.19-7.13(m,3H),6.37(s,2H),5.95(d,J＝5.5Hz,1H),5.36(m,1H),4.91-4.88(m,1H),3.97-3.84(m,2H),3.71-3.63(m,1H)，3.21-3.02(m,3H),2.6-2.59(m,1H),1.89-1.75(m,2H),1.56-1.53(m,1H),1.39(d,J＝6.0Hz,3H),1.17-1.13(m,1H). 1 H NMR(500MHz, DMSO-d 6 )δ8.32(s,1H), 7.67(d,J=5.5Hz,1H), 7.31(d,J=7.0Hz,1H), 7.19-7.13(m, 3H), 6.37 (s, 2H), 5.95 (d, J = 5.5 Hz, 1H), 5.36 (m, 1H), 4.91-4.88 (m, 1H), 3.97-3.84 (m, 2H), 3.71-3.63 (m,1H),3.21-3.02(m,3H),2.6-2.59(m,1H),1.89-1.75(m,2H),1.56-1.53(m,1H),1.39(d,J=6.0Hz ,3H),1.17-1.13(m,1H).
经由不同的反应起始原料和合适的试剂，采用与前述实施例26类似的方法制备表3中的化合物28-57及63。The compounds 28-57 and 63 in Table 3 were prepared by using different reaction starting materials and appropriate reagents, using a method similar to that of the foregoing Example 26.
表3table 3
Figure PCTCN2020072774-appb-000051
Figure PCTCN2020072774-appb-000051
Figure PCTCN2020072774-appb-000052
Figure PCTCN2020072774-appb-000052
Figure PCTCN2020072774-appb-000053
Figure PCTCN2020072774-appb-000053
Figure PCTCN2020072774-appb-000054
Figure PCTCN2020072774-appb-000054
化合物30、33、35、36、37、41、42、45、46、47的核磁数据如下：The NMR data of compounds 30, 33, 35, 36, 37, 41, 42, 45, 46, 47 are as follows:
[M+H +]＝467.25 [M+H + ]=467.25
1H NMR(500MHz,DMSO-d 6):δ8.25(s,1H),7.67(d,J＝5.4Hz,1H),7.31(d,J＝6.9Hz,1H),7.20-7.12(m,3H),6.69(d,J＝5.1Hz,1H),6.12(t,J＝4.5Hz,1H),5.46(s,1H),4.50(s,2H),3.85-3.77(m,3H),3.18-3.11(m,2H),3.03(d,J＝15.6Hz,1H),2.82(d,J＝4.5Hz,3H),2.61(d,J＝15.6Hz,1H),1.88(m,1H),1.83-1.74(m,1H),1.55-1.52(m,1H),1.14-1.11(m,1H).(化合物30) 1 H NMR (500MHz, DMSO-d 6 ): δ8.25 (s, 1H), 7.67 (d, J = 5.4 Hz, 1H), 7.31 (d, J = 6.9 Hz, 1H), 7.20-7.12 (m ,3H),6.69(d,J=5.1Hz,1H),6.12(t,J=4.5Hz,1H),5.46(s,1H),4.50(s,2H),3.85-3.77(m,3H) ,3.18-3.11(m,2H),3.03(d,J=15.6Hz,1H), 2.82(d,J=4.5Hz,3H), 2.61(d,J=15.6Hz,1H),1.88(m, 1H), 1.83-1.74 (m, 1H), 1.55-1.52 (m, 1H), 1.14-1.11 (m, 1H). (Compound 30)
[M+H +]＝467.26 [M+H + ]=467.26
1H NMR(500MHz,DMSO-d 6):δ8.11(s,1H),7.31(d,J＝6.5Hz,1H),7.25-7.09(m,4H),7.08-7.04(m,1H),6.89(d,J＝7.8Hz,1H),5.39(t,J＝5.6Hz,1H),4.50(d,J＝5.5Hz,2H),3.85(s,3H),3.75-3.70(m,2H),3.14-3.02(m,3H),2.61(d,J＝15.6Hz,2H),1.91-1.81(m,1H),1.81-1.76(m,1H),1.53(d,J＝12.9Hz,1H),1.13(d,J＝13.3Hz,1H).(化合物33) 1 H NMR (500MHz, DMSO-d 6 ): δ8.11 (s, 1H), 7.31 (d, J = 6.5 Hz, 1H), 7.25-7.09 (m, 4H), 7.08-7.04 (m, 1H) ,6.89(d,J=7.8Hz,1H),5.39(t,J=5.6Hz,1H),4.50(d,J=5.5Hz,2H),3.85(s,3H),3.75-3.70(m, 2H),3.14-3.02(m,3H),2.61(d,J=15.6Hz,2H),1.91-1.81(m,1H),1.81-1.76(m,1H),1.53(d,J=12.9Hz ,1H),1.13(d,J=13.3Hz,1H). (Compound 33)
[M+H +]＝470.32 [M+H + ]=470.32
1H NMR(500MHz,DMSO-d 6):δ8.66(d,J＝4.7Hz,1H),8.36(s,1H),8.20(d,J＝8.3Hz,1 H),8.05(d,J＝8.4Hz,1H),7.84(t,J＝7.6Hz,1H),7.71(t,J＝7.5Hz,1H),7.32(d,J＝6.6Hz,1H),7.20-7.16(m,3H),7.04(d,J＝4.6Hz,1H),5.49(t,J＝5.8Hz,1H),4.54(d,J＝5.7Hz,2H),4.01-3.80(m,3H),3.21-3.18(m,2H),3.07(d,J＝15.7Hz,1H),2.62(d,J＝15.7Hz,1H),1.93-1.88(m,1H),1.83-1.78(m,1H),1.57-1.54(m,1H),1.17-1.13(m,1H).(化合物35) 1 H NMR (500MHz, DMSO-d 6 ): δ8.66(d,J=4.7Hz,1H), 8.36(s,1H), 8.20(d,J=8.3Hz,1H), 8.05(d, J = 8.4 Hz, 1H), 7.84 (t, J = 7.6 Hz, 1H), 7.71 (t, J = 7.5 Hz, 1H), 7.32 (d, J = 6.6 Hz, 1H), 7.20-7.16 (m, 3H), 7.04 (d, J = 4.6 Hz, 1H), 5.49 (t, J = 5.8 Hz, 1H), 4.54 (d, J = 5.7 Hz, 2H), 4.01-3.80 (m, 3H), 3.21- 3.18(m,2H),3.07(d,J=15.7Hz,1H), 2.62(d,J=15.7Hz,1H),1.93-1.88(m,1H),1.83-1.78(m,1H),1.57 -1.54 (m, 1H), 1.17-1.13 (m, 1H). (Compound 35)
[M+H] +:488.3 [M+H] + :488.3
1H NMR(500MHz,DMSO-d 6)δ:8.58-8.56(m,1H),8.25(s,1H),7.81-7.65(m,1H),7.62-7.60(m,1H),7.32-7.29(m,1H),7.17-7.13(m,3H),5.41(t,J＝5.6Hz,1H),4.49-4.47(m,2H),3.85–3.75(m,2H),3.20-3.10(m,2H),3.05-3.02(m,1H),2.63-2.55(m,1H),1.96-1.67(m,1H),1.55-1.52(m,1H),1.18-1.08(m,1H).(化合物36) 1 H NMR (500MHz, DMSO-d 6 ) δ: 8.58-8.56 (m, 1H), 8.25 (s, 1H), 7.81-7.65 (m, 1H), 7.62-7.60 (m, 1H), 7.32-7.29 (m,1H),7.17-7.13(m,3H),5.41(t,J=5.6Hz,1H),4.49-4.47(m,2H),3.85-3.75(m,2H),3.20-3.10(m ,2H),3.05-3.02(m,1H),2.63-2.55(m,1H),1.96-1.67(m,1H),1.55-1.52(m,1H),1.18-1.08(m,1H). Compound 36)
[M+H] +:487.17 [M+H] + :487.17
1H NMR(500MHz,DMSO-d 6)δ:8.24(s,1H),7.51(d,J＝8.0Hz,1H),7.34–7.25(m,2H),7.17(q,J＝7.6,7.2Hz,3H),6.99(d,J＝8.0Hz,1H),5.43(t,J＝5.8Hz,1H),4.51(d,J＝5.8Hz,2H),3.85-3.77(m,3H),3.20-3.10(m,2H),3.05(d,J＝15.6Hz,1H),2.62(d,J＝15.6Hz,1H),2.02-1.68(m,4H),1.55-1.53(m,1H),1.15-1.13(m,1H).(化合物37) 1 H NMR (500MHz, DMSO-d 6 ) δ: 8.24 (s, 1H), 7.51 (d, J = 8.0 Hz, 1H), 7.34-7.25 (m, 2H), 7.17 (q, J = 7.6, 7.2 Hz, 3H), 6.99 (d, J = 8.0 Hz, 1H), 5.43 (t, J = 5.8 Hz, 1H), 4.51 (d, J = 5.8 Hz, 2H), 3.85-3.77 (m, 3H), 3.20-3.10(m,2H),3.05(d,J=15.6Hz,1H), 2.62(d,J=15.6Hz,1H), 2.02-1.68(m,4H), 1.55-1.53(m,1H) ,1.15-1.13(m,1H).(Compound 37)
[M+H] +:487.21 [M+H] + :487.21
1H NMR(500MHz,DMSO-d 6)δ8.31(s,1H),7.67(d,J＝5.6Hz,1H),7.21-7.18(m,1H),7.09(d,J＝9.0Hz,1H),6.95(t,J＝8.8Hz,1H),6.36(s,2H),5.99-5.88(m,1H),5.49(t,J＝5.9Hz,1H),4.52(d,J＝5.8Hz,2H),3.92-3.85(m,3H),3.21-3.01(m,3H),2.63-2.57(m,1H),1.99-1.89(m,1H),1.84-1.74(m,1H),1.58-1.55(m,1H),1.13-1.10(m,1H).(化合物41) 1 H NMR(500MHz,DMSO-d 6 )δ8.31(s,1H), 7.67(d,J=5.6Hz,1H), 7.21-7.18(m,1H), 7.09(d,J=9.0Hz, 1H), 6.95(t,J=8.8Hz,1H), 6.36(s,2H),5.99-5.88(m,1H), 5.49(t,J=5.9Hz,1H),4.52(d,J=5.8 Hz,2H),3.92-3.85(m,3H),3.21-3.01(m,3H),2.63-2.57(m,1H),1.99-1.89(m,1H),1.84-1.74(m,1H), 1.58-1.55 (m, 1H), 1.13-1.10 (m, 1H). (Compound 41)
[M+H] +:499.23 [M+H] + :499.23
1H NMR(500MHz,DMSO-d 6)δ8.31(s,1H),7.67(d,J＝5.4Hz,1H),7.07(d,J＝8.2Hz,1H),6.90(m,1H),6.70(d,J＝7.9Hz,1H),6.36(s,2H),5.91(d,J＝5.3Hz,1H),5.48(m,1H),4.52(d,J＝4.9Hz,2H),3.94-3.76(m,3H),3.73(s,3H),3.20-3.10(m,2H),2.98-2.95(m,1H),2.64-2.55(m,1H),1.93-1.88(m,1H),1.80-1.75(m,1H),1.56-1.53(m,1H),1.24-1.11(m,1H).(化合物42) 1 H NMR(500MHz,DMSO-d 6 )δ8.31(s,1H), 7.67(d,J=5.4Hz,1H), 7.07(d,J=8.2Hz,1H), 6.90(m,1H) ,6.70(d,J=7.9Hz,1H),6.36(s,2H),5.91(d,J=5.3Hz,1H),5.48(m,1H),4.52(d,J=4.9Hz,2H) ,3.94-3.76(m,3H),3.73(s,3H),3.20-3.10(m,2H),2.98-2.95(m,1H),2.64-2.55(m,1H),1.93-1.88(m, 1H), 1.80-1.75 (m, 1H), 1.56-1.53 (m, 1H), 1.24-1.11 (m, 1H). (Compound 42)
[M+H] +:494.21 [M+H] + :494.21
1H NMR(500MHz,DMSO-d 6)δ8.32(s,1H),7.69-7.61(m,3H),7.41(t,J＝7.6Hz,1H),6.35(s,2H),5.92(d,J＝5.4Hz,1H),5.49(t,J＝5.8Hz,1H),4.53(d,J＝5.8Hz,2H),4.06-3.85(m,3H),3.29-3.18(m,3H),2.86-2.83(m,1H),1.91-1.80(m,2H),1.61-1.59(m,1H),1.22–1.15(m,1H).(化合物45) 1 H NMR(500MHz,DMSO-d 6 )δ8.32(s,1H),7.69-7.61(m,3H),7.41(t,J=7.6Hz,1H),6.35(s,2H),5.92( d,J=5.4Hz,1H),5.49(t,J=5.8Hz,1H),4.53(d,J=5.8Hz,2H),4.06-3.85(m,3H),3.29-3.18(m,3H ), 2.86-2.83 (m, 1H), 1.91-1.80 (m, 2H), 1.61-1.59 (m, 1H), 1.22-1.15 (m, 1H). (Compound 45)
[M+H] +:503.14 [M+H] + :503.14
1H NMR(500MHz,DMSO-d 6)δ8.31(s,1H),7.67(d,J＝6.0Hz,1H),7.32(s,1H),7.24–7.16(m,2H),6.36(s,2H),5.91(d,J＝5.3Hz,1H),5.49(t,J＝5.9Hz,1H),4.52(d,J＝5.8Hz,2H),3.92-3.86(m,3H),3.21-3.11(m,2H),3.05(d,J＝15.7Hz,1H),2.60(d,J＝15.7Hz,1H),1.93-1.88(m,1H),1.80-1.75(m,1H),1.56(m,1H),1.12-1.10(m,1H).(化合物46) 1 H NMR(500MHz,DMSO-d 6 )δ8.31(s,1H), 7.67(d,J=6.0Hz,1H), 7.32(s,1H), 7.24–7.16(m,2H), 6.36( s, 2H), 5.91 (d, J = 5.3 Hz, 1H), 5.49 (t, J = 5.9 Hz, 1H), 4.52 (d, J = 5.8 Hz, 2H), 3.92-3.86 (m, 3H), 3.21-3.11(m,2H),3.05(d,J=15.7Hz,1H), 2.60(d,J=15.7Hz,1H),1.93-1.88(m,1H),1.80-1.75(m,1H) , 1.56 (m, 1H), 1.12-1.10 (m, 1H). (Compound 46)
[M+H] +:420.26 [M+H] + :420.26
1H NMR(500MHz,DMSO-d 6)δ8.41-8.36(m,2H),8.32(s,1H),7.32(d,J＝6.8Hz,1H),7.23-7.09(m,5H),5.48(t,J＝5.6Hz,1H),4.53(d,J＝5.6Hz,2H),3.91-3.80(m,3H),3.21-3.13(m,2H),3.06(d,J＝15.6Hz,1H),2.65(d,J＝15.6Hz,1H),1.93-1.77(m,2H),1.55–1.53(m,1H),1.18-1.15(m,1H).(化合物47) 1 H NMR(500MHz,DMSO-d 6 )δ8.41-8.36(m,2H),8.32(s,1H),7.32(d,J=6.8Hz,1H),7.23-7.09(m,5H), 5.48(t,J=5.6Hz,1H),4.53(d,J=5.6Hz,2H),3.91-3.80(m,3H),3.21-3.13(m,2H),3.06(d,J=15.6Hz ,1H), 2.65(d,J=15.6Hz,1H),1.93-1.77(m,2H),1.55-1.53(m,1H),1.18-1.15(m,1H). (Compound 47)
实施例60化合物60的制备：Example 60 Preparation of Compound 60:
Figure PCTCN2020072774-appb-000055
Figure PCTCN2020072774-appb-000055
步骤1：化合物60-1的制备Step 1: Preparation of compound 60-1
将13mg化合物21-2A溶解于0.5mL MeOH中，滴加3mg LiOH·H 2O的H 2O(0.1mL)溶液，反应液升温至40℃搅拌反应12hrs。TLC检测反应完全，用1N HCl溶液调节反应液至pH＝7，加DCM/MeOH＝10/1萃取(5mL×2)，合并有机相，用5mL饱和NaCl洗涤，无水硫酸钠干燥，减压浓缩，残余物经TLC板纯化得10mg化合物60-1。 13 mg of compound 21-2A was dissolved in 0.5 mL of MeOH, 3 mg of LiOH·H 2 O in H 2 O (0.1 mL) was added dropwise, and the reaction solution was heated to 40° C. and stirred for 12 hrs. TLC detects the completion of the reaction, adjust the reaction solution to pH=7 with 1N HCl solution, add DCM/MeOH=10/1 to extract (5mL×2), combine the organic phases, wash with 5mL saturated NaCl, dry with anhydrous sodium sulfate, and reduce pressure After concentration, the residue was purified by TLC plate to obtain 10 mg of compound 60-1.
步骤2：化合物60-3的制备Step 2: Preparation of compound 60-3
将10mg化合物60-1溶解于0.3mL THF和0.3mL DCM中，加入4mg EDCI，3mg HOBT和2mg TEA，RT搅拌反应1hrs。再加入3mg化合物60-2。氮气置换三次，氮气保护下RT搅拌反应12hrs。TLC检测反应完全，减压浓缩，残余物经TLC板纯化得6mg化合物60-3。Dissolve 10 mg of compound 60-1 in 0.3 mL of THF and 0.3 mL of DCM, add 4 mg of EDCI, 3 mg of HOBT, and 2 mg of TEA, and stir for 1 hrs at RT. An additional 3 mg of compound 60-2 was added. Replace with nitrogen for three times, and stir for 12hrs at RT under nitrogen protection. TLC detected that the reaction was complete, concentrated under reduced pressure, and the residue was purified by TLC plate to obtain 6 mg of compound 60-3.
步骤3：化合物60的制备Step 3: Preparation of compound 60
将6mg化合物60-3溶解于0.5mL MeOH中，加入2N HCl(0.1mL，甲醇溶液)，RT搅拌反应8hrs。TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(0.5mL)洗涤，取滤饼真空干燥得2.2mg黄色固体化合物60。 6mg of compound 60-3 was dissolved in 0.5mL MeOH, 2N HCl (0.1mL, methanol solution) was added, and the reaction was stirred at RT for 8hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H 2 O (0.5 mL) to dissolve the residue, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (0.5 mL), take the filter cake and vacuum dry to obtain 2.2 mg of yellow solid Compound 60.
[M+H +]＝498.23。 [M+H + ]=498.23.
实施例61化合物61的制备：Example 61 Preparation of Compound 61:
Figure PCTCN2020072774-appb-000056
Figure PCTCN2020072774-appb-000056
步骤1：化合物61-3的制备Step 1: Preparation of compound 61-3
将100mg化合物M23-1和57mg化合物61-2溶解于2mL四氢呋喃中，加入0.01ml四丁基氟化氨,然后RT反应2小时。TLC检测原料反应完全，将反应液旋干，经柱层析纯化得61mg化合物61-3。100mg of compound M23-1 and 57mg of compound 61-2 were dissolved in 2mL of tetrahydrofuran, 0.01ml of tetrabutylammonium fluoride was added, and then reacted for 2 hours at RT. TLC detected the complete reaction of the raw material, the reaction solution was spin-dried, and purified by column chromatography to obtain 61 mg of compound 61-3.
步骤2：化合物61-4的制备Step 2: Preparation of compound 61-4
将61mg化合物61-3、50mg化合物M1溶解于1.5mL的二氧六环中，加入10mg Pd 2(dba) 3、12mg Xantphos和42mg DIEA，然后100℃反应4hrs。LCMS检测反应完全，将反应液过滤，减压浓缩，残余物用制备板纯化得35mg白色固体化合物61-4。 61 mg of compound 61-3 and 50 mg of compound M1 were dissolved in 1.5 mL of dioxane, 10 mg Pd 2 (dba) 3 , 12 mg Xantphos, and 42 mg DIEA were added, and then reacted at 100° C. for 4 hrs. LCMS detected that the reaction was complete, the reaction solution was filtered, concentrated under reduced pressure, and the residue was purified with a preparation plate to obtain 35 mg of white solid compound 61-4.
步骤3：化合物61的制备Step 3: Preparation of compound 61
将35mg化合物61-4，溶解于1.5mL的二氧六环和0.5ml甲醇中，加入0.14mL HCl的二氧六环溶液，室温下搅拌2小时。LCMS检测反应完全，将反应液减压浓缩，残余物用1mL水溶解，滴加饱和碳酸氢钠溶液调pH＝8，析出固体，过滤烘干得20.6mg白色固体化合物61。Dissolve 35 mg of compound 61-4 in 1.5 mL of dioxane and 0.5 mL of methanol, add 0.14 mL of HCl in dioxane solution, and stir at room temperature for 2 hours. LCMS detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, the residue was dissolved in 1 mL of water, saturated sodium bicarbonate solution was added dropwise to adjust pH=8, a solid was precipitated, filtered and dried to obtain 20.6 mg of white solid compound 61.
[M+H +]＝537.18。 [M+H + ]=537.18.
实施例62化合物62的制备：Example 62 Preparation of Compound 62:
Figure PCTCN2020072774-appb-000057
Figure PCTCN2020072774-appb-000057
步骤1：化合物62-2的制备Step 1: Preparation of compound 62-2
将100mg化合物M22-2、72mg的苄基三丁基溴化铵溶解于4mL氢氧化钠(50％)和4mL二氯甲烷的混合溶液中。0℃下滴加26mg化合物62-1，缓慢升至室温。TLC检测反应完全，加入30mL水淬灭反应，再用EA(50mL)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得49mg化合物62-2。100 mg of compound M22-2 and 72 mg of benzyl tributylammonium bromide were dissolved in a mixed solution of 4 mL of sodium hydroxide (50%) and 4 mL of dichloromethane. 26 mg of compound 62-1 was added dropwise at 0°C and slowly warmed to room temperature. TLC detected that the reaction was complete. The reaction was quenched by adding 30 mL of water, and then extracted with EA (50 mL). The organic layers were combined, dried over sodium sulfate and then desolventized, and purified by column chromatography to obtain 49 mg of compound 62-2.
步骤2：化合物62-3的制备Step 2: Preparation of compound 62-3
将49mg化合物62-2、53mg化合物M1、25mg DIEA、9mg Pd 2(dba) 3和10mg Xantphos溶解于2mL二氧六环中，氮气置换3次后，100℃反应3hrs。停止加热，减压浓缩，加入20mL水，再用EA(30mL×2)萃取，合并有机层，用硫酸钠干燥后脱溶，经柱层析纯化得27mg化合物62-3。 49 mg of compound 62-2, 53 mg of compound M1, 25 mg of DIEA, 9 mg of Pd 2 (dba) 3 and 10 mg of Xantphos were dissolved in 2 mL of dioxane, replaced with nitrogen three times, and reacted at 100°C for 3 hrs. Stop heating, concentrate under reduced pressure, add 20 mL of water, and extract with EA (30 mL×2). Combine the organic layers, dry with sodium sulfate and desolvate, and purify by column chromatography to obtain 27 mg of compound 62-3.
步骤3：化合物62的制备Step 3: Preparation of compound 62
将27mg化合物62-3，溶于1mL二氧六环中，滴加盐酸甲醇溶液(2N,0.4mL)，TLC跟踪反应。反应完全后，减压除去溶剂，用少量正己烷固化后，倾倒掉正己烷，加入0.5mL水溶解固体，再滴入2滴饱和碳酸氢钠水溶液。过滤得到固体，固体用少量水洗涤后，真空干燥。得到10.5mg淡黄色固体化合物62。27 mg of compound 62-3 was dissolved in 1 mL of dioxane, and methanol solution of hydrochloric acid (2N, 0.4 mL) was added dropwise, and the reaction was followed by TLC. After the reaction was completed, the solvent was removed under reduced pressure, and after solidification with a small amount of n-hexane, the n-hexane was poured out, 0.5 mL of water was added to dissolve the solid, and 2 drops of saturated sodium bicarbonate aqueous solution were added dropwise. The solid was obtained by filtration, and the solid was washed with a small amount of water and dried under vacuum. 10.5 mg of compound 62 was obtained as a pale yellow solid.
1H NMR(500MHz,DMSO-d 6,):δ8.35(s,1H),7.67(d,J＝5.4Hz,1H),7.31(d,J＝6.5Hz,1H),7.18-7.15(m,3H),6.36(s,2H),5.88(d,J＝5.1Hz,1H),4.45(s,2H),3.93-3.86(m,3H),3.25-3.15(m,2H),3.06(d,J＝15.7Hz,1H),2.63(d,J＝15.3Hz,1H),1.92-1.88(m,1H),1.82-1.78(m,1H),1.55(d,J＝12.9Hz,1H),1.16(d,J＝13.8Hz,1H)。 1 H NMR (500MHz, DMSO-d 6 ,): δ8.35(s,1H), 7.67(d,J=5.4Hz,1H), 7.31(d,J=6.5Hz,1H), 7.18-7.15( m,3H),6.36(s,2H),5.88(d,J=5.1Hz,1H),4.45(s,2H),3.93-3.86(m,3H),3.25-3.15(m,2H),3.06 (d,J=15.7Hz,1H), 2.63(d,J=15.3Hz,1H),1.92-1.88(m,1H),1.82-1.78(m,1H),1.55(d,J=12.9Hz, 1H), 1.16 (d, J=13.8 Hz, 1H).
实施例64化合物64的制备：Example 64 Preparation of Compound 64:
Figure PCTCN2020072774-appb-000058
Figure PCTCN2020072774-appb-000058
步骤1：化合物64-1的制备Step 1: Preparation of compound 64-1
将400mg化合物M22-SM和552mg化合物M12溶解于8.0mL THF中，加入349mg N,N-二异丙基乙胺，40℃搅拌反应16hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得420mg浅黄色固体化合物64-1。400 mg of compound M22-SM and 552 mg of compound M12 were dissolved in 8.0 mL of THF, 349 mg of N,N-diisopropylethylamine was added, and the reaction was stirred at 40°C for 16 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 420 mg of light yellow solid compound 64-1.
步骤2：化合物64-2的制备Step 2: Preparation of compound 64-2
将420mg化合物64-1溶解于5mL的无水DCM中。在氮气保护下，-78℃缓慢滴加3.8mL1.0M DIBAL-H的正己烷溶液，-78℃搅拌反应15min。LCMS检测原料反应完全，低温下依次滴加0.5mL水、0.5mL NaOH(2.5M)水溶液淬灭，回温至室温再滴加0.5mL水，过滤后有机相无水硫酸钠干燥，减压浓缩得385mg粗品化合物64-2。420 mg of compound 64-1 was dissolved in 5 mL of anhydrous DCM. Under the protection of nitrogen, slowly drop 3.8 mL 1.0M DIBAL-H n-hexane solution at -78°C, and stir at -78°C for 15 min. LCMS detects that the reaction of the raw materials is complete. 0.5mL water and 0.5mL NaOH (2.5M) aqueous solution are added dropwise to quench at low temperature. After warming to room temperature, 0.5mL water is added dropwise. After filtration, the organic phase is dried over anhydrous sodium sulfate and concentrated under reduced pressure. 385 mg of crude compound 64-2 was obtained.
步骤3：化合物64-3的制备Step 3: Preparation of compound 64-3
将385mg化合物64-2溶解于5mL无水THF中。在氮气保护下，0℃加入56mg NaBH 4，恢复室温搅拌反应2hrs。LCMS检测反应完全，冰浴下滴加入5mL饱和氯化铵，恢复室温，真空浓缩旋干THF后，水相用DCM(5mL×3)萃取，合并有机相，无水硫酸钠干燥，减压浓缩，残余物经Pre-TLC纯化得130mg黄色固体化合物64-3。 385 mg of compound 64-2 was dissolved in 5 mL of anhydrous THF. Under the protection of nitrogen, 56mg NaBH 4 was added at 0°C, and the reaction was stirred at room temperature for 2hrs. LCMS detected that the reaction was complete, 5mL saturated ammonium chloride was added dropwise under ice bath, returned to room temperature, concentrated in vacuo and spin-dried THF, the aqueous phase was extracted with DCM (5mL×3), the organic phases were combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure The residue was purified by Pre-TLC to obtain 130 mg of yellow solid compound 64-3.
步骤4：化合物64-4的制备Step 4: Preparation of compound 64-4
将130mg化合物64-3和63mg化合物22-3溶解于5mL 1,4-二氧六环中，加入12mg Pd 2(dba) 3、15mg Xantphos和65mg N,N-二异丙基乙胺。氮气置换三次，氮气保护下反应升温至70℃搅拌反应2hrs。LCMS和TLC检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经Pre-TLC纯化得70mg黄色固体化合物64-4。 130 mg of compound 64-3 and 63 mg of compound 22-3 were dissolved in 5 mL of 1,4-dioxane, 12 mg of Pd 2 (dba) 3 , 15 mg of Xantphos and 65 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was heated to 70°C and stirred for 2hrs. LCMS and TLC detected the completion of the reaction, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by Pre-TLC to obtain 70 mg of yellow solid compound 64-4.
步骤4：化合物64的制备Step 4: Preparation of compound 64
将70mg化合物64-4溶解于3.0mL1,4-二氧六环和0.6mL MeOH中，加入2N HCl(0.3mL，甲醇溶液)，室温搅拌反应2.0hrs。LCMS和TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8～9，固体析出，过滤，滤饼固体 再用DCM溶解，加入无水硫酸钠干燥后，过滤真空浓缩，得50.2mg浅黄色固体化合物64。 70 mg of compound 64-4 was dissolved in 3.0 mL 1,4-dioxane and 0.6 mL MeOH, 2N HCl (0.3 mL, methanol solution) was added, and the reaction was stirred at room temperature for 2.0 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. The residue was dissolved by adding H 2 O (0.5 mL), adjusting the solution to pH=8-9 with saturated NaHCO 3 , solids separated out, filtered, the filter cake solid was dissolved in DCM, dried by adding anhydrous sodium sulfate, filtered and concentrated in vacuo. 50.2 mg of compound 64 was obtained as a light yellow solid.
[M+H +]＝466.34。 [M+H + ]=466.34.
1H NMR(500MHz,DMSO-d 6)δ8.33(s,1H),7.96(d,J＝5.0Hz,1H),7.08(d,J＝8.0Hz,1H),6.90-6.89(m,1H),6.72-6.69(m,3H),6.09(d,J＝5.5Hz,1H),5.50(m,1H),4.52(d,J＝2.5Hz,2H),3.89-3.81(m,3H),3.73(s,3H),3.18-3.08(m,2H),2.96(d,J＝15.0Hz,1H),2.54(d,J＝15.0Hz,1H),1.93-1.88(m,1H),1.80-1.75(m,1H),1.55-1.53(m,1H),1.13-1.10(m,1H)。 1 H NMR(500MHz,DMSO-d 6 )δ8.33(s,1H), 7.96(d,J=5.0Hz,1H), 7.08(d,J=8.0Hz,1H), 6.90-6.89(m, 1H), 6.72-6.69 (m, 3H), 6.09 (d, J = 5.5 Hz, 1H), 5.50 (m, 1H), 4.52 (d, J = 2.5 Hz, 2H), 3.89-3.81 (m, 3H) ),3.73(s,3H),3.18-3.08(m,2H),2.96(d,J=15.0Hz,1H),2.54(d,J=15.0Hz,1H),1.93-1.88(m,1H) , 1.80-1.75 (m, 1H), 1.55-1.53 (m, 1H), 1.13-1.10 (m, 1H).
实施例65化合物65的制备：Example 65 Preparation of compound 65:
Figure PCTCN2020072774-appb-000059
Figure PCTCN2020072774-appb-000059
步骤1：化合物65-1的制备Step 1: Preparation of compound 65-1
将150mg化合物M22-2和41mg化合物4-巯基嘧啶溶解于5mL 1,4-二氧六环中，加入14mg Pd 2(dba) 3、18mg Xantphos和118mg N,N-二异丙基乙胺。氮气置换三次，氮气保护下反应升温至70℃搅拌反应2hrs。LCMS和TLC检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经Pre-TLC纯化得60mg黄色固体化合物65-1。 150 mg of compound M22-2 and 41 mg of compound 4-mercaptopyrimidine were dissolved in 5 mL of 1,4-dioxane, and 14 mg of Pd 2 (dba) 3 , 18 mg of Xantphos and 118 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was heated to 70°C and stirred for 2hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by Pre-TLC to obtain 60 mg of yellow solid compound 65-1.
步骤2：化合物65的制备Step 2: Preparation of compound 65
将60mg化合物65-1溶解于1.5mL 1,4-二氧六环和0.5mL MeOH中，加入2N HCl(0.3mL，甲醇溶液)，室温搅拌反应1.0hrs。LCMS和TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝7-8，析出固体，过滤，滤饼水洗，烘干得19.1mg浅黄色固体65。 60 mg of compound 65-1 was dissolved in 1.5 mL 1,4-dioxane and 0.5 mL MeOH, 2N HCl (0.3 mL, methanol solution) was added, and the reaction was stirred at room temperature for 1.0 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. The residue was dissolved by adding H 2 O (0.5 mL), the solution was adjusted to pH=7-8 with saturated NaHCO 3 , a solid was precipitated, filtered, the filter cake was washed with water, and dried to obtain 19.1 mg of light yellow solid 65.
[M+H +]＝421.27。 [M+H + ]=421.27.
1H NMR(500MHz,DMSO-d 6)δ8.91(s,1H),8.51(d,J＝5.5Hz,1H),8.36(s,1H),7.32(d,J＝6.5Hz,1H),7.21-7.13(m,4H),5.30(t,J＝6.0Hz,1H),4.53(s,2H),3.93-3.86(m,3H),3.33-3.16(m,2H),3.07(d,J＝15.0Hz,1H),2.64(d,J＝15.5Hz,1H),1.93-1.88(m,1H),1.82-1.77(m,1H),1.57-1.54(m,1H),1.16-1.13(m,1H)。 1 H NMR(500MHz,DMSO-d 6 )δ8.91(s,1H),8.51(d,J=5.5Hz,1H),8.36(s,1H),7.32(d,J=6.5Hz,1H) ,7.21-7.13(m,4H),5.30(t,J=6.0Hz,1H),4.53(s,2H),3.93-3.86(m,3H),3.33-3.16(m,2H),3.07(d ,J=15.0Hz,1H),2.64(d,J=15.5Hz,1H),1.93-1.88(m,1H),1.82-1.77(m,1H),1.57-1.54(m,1H),1.16- 1.13(m,1H).
实施例66化合物66的制备：Example 66 Preparation of compound 66:
Figure PCTCN2020072774-appb-000060
Figure PCTCN2020072774-appb-000060
步骤1：化合物66-1的制备Step 1: Preparation of compound 66-1
将500mg化合物M22-SM和634mg化合物M16溶解于9.0mL二氧六环中，加入655mg N,N-二异丙基乙胺。70℃搅拌反应2hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得444mg黄色固体化合物66-1。500 mg of compound M22-SM and 634 mg of compound M16 were dissolved in 9.0 mL of dioxane, and 655 mg of N,N-diisopropylethylamine was added. The reaction was stirred at 70°C for 2hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 444 mg of yellow solid compound 66-1.
步骤2：化合物66-2的制备Step 2: Preparation of compound 66-2
将444mg化合物66-1溶解于10mL的无水DCM中。在氮气保护下，-78℃缓慢滴加4.0mL 1.0M DIBAL-H的正己烷溶液，-78℃搅拌反应0.5hrs。LCMS检测反应完全，低温下依次滴加0.5mL水，0.5mL 2.5M NaOH水溶液淬灭，恢复室温再滴加0.5mL水，过滤后有机相无水硫酸钠干燥，减压浓缩得380mg粗品化合物66-2。444 mg of compound 66-1 was dissolved in 10 mL of anhydrous DCM. Under the protection of nitrogen, slowly add 4.0mL 1.0M DIBAL-H n-hexane solution dropwise at -78℃, and stir for 0.5hrs at -78℃. The reaction was completed by LCMS, 0.5mL water and 0.5mL 2.5M NaOH aqueous solution were added dropwise to quench at low temperature, and 0.5mL water was added dropwise after returning to room temperature. After filtration, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 380mg of crude compound 66 -2.
步骤3：化合物66-3的制备Step 3: Preparation of compound 66-3
将380mg化合物66-2溶解于10mL的无水THF中。在氮气保护下，0℃加入60mg NaBH 4，恢复室温搅拌反应0.5hrs。LCMS检测反应完全，冰浴下滴加入5mL水淬灭，恢复室温，真空浓缩旋干THF后，水相用DCM(5mL×3)萃取，合并有机相无水硫酸钠干燥，减压浓缩，残余物经柱层析纯化得300mg黄色固体化合物66-3。 380 mg of compound 66-2 was dissolved in 10 mL of anhydrous THF. Under the protection of nitrogen, 60mg NaBH 4 was added at 0°C, and the reaction was stirred at room temperature for 0.5 hrs. LCMS detected the completion of the reaction. The reaction was quenched by adding 5 mL of water dropwise under an ice bath. The temperature was restored to room temperature. After vacuum concentration and spin-drying of THF, the aqueous phase was extracted with DCM (5 mL×3), and the combined organic phases were dried over anhydrous sodium sulfate and concentrated under reduced pressure. The product was purified by column chromatography to obtain 300 mg of yellow solid compound 66-3.
步骤4：化合物66-4的制备Step 4: Preparation of compound 66-4
将200mg化合物66-3和96mg化合物22-3溶解于5mL二氧六环中，加入35mg Pd 2(dba) 3，44mg Xantphos、147mg N,N-二异丙基乙胺。氮气置换三次，氮气保护下反应升至70℃搅拌反应2hrs。LCMS和TLC检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得160mg化合物66-4。 200 mg of compound 66-3 and 96 mg of compound 22-3 were dissolved in 5 mL of dioxane, and 35 mg of Pd 2 (dba) 3 , 44 mg of Xantphos, and 147 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction temperature is raised to 70°C and the reaction is stirred for 2hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 160 mg of compound 66-4.
步骤5：化合物66的制备Step 5: Preparation of compound 66
将50mg化合物66-4溶解于1.5mL二氧六环和0.3mL MeOH中，加入2N HCl(0.5mL，甲醇溶液)，RT搅拌反应0.5hrs。LCMS和TLC检测反应完全，反应液减压浓缩。残余物加入 H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(0.5mL)洗涤，固体再用DCM溶解，干燥后真空浓缩，经制备板纯化得5.8mg白色固体66。 50 mg of compound 66-4 was dissolved in 1.5 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.5 mL, methanol solution) was added, and the reaction was stirred at RT for 0.5 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H 2 O (0.5 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (0.5 mL), dissolve the solid with DCM, dry and concentrate in vacuo , Purified by preparation plate to obtain 5.8 mg of white solid 66.
[M+H +]＝470.31。 [M+H + ]=470.31.
1H NMR(500MHz,DMSO-d 6)δ8.34(s,1H),7.96(d,J＝5.0Hz,1H),7.32(d,J＝8.0Hz,1H),7.24-7.21(m,2H),6.72(s,2H),6.08(d,J＝5.5Hz,1H),5.50(t,J＝6.0Hz,1H),4.52(d,J＝6.0Hz,2H),3.88-3.82(m,3H),3.18-3.09(m,2H),3.06(d,J＝16.0Hz,1H),2.63(d,J＝15.5Hz,1H),1.92-1.86(m,1H),1.81-1.76(m,1H),1.56-1.53(m,1H),1.14-1.13(m,1H)。 1 H NMR(500MHz,DMSO-d 6 )δ8.34(s,1H),7.96(d,J=5.0Hz,1H), 7.32(d,J=8.0Hz,1H), 7.24-7.21(m, 2H), 6.72 (s, 2H), 6.08 (d, J = 5.5 Hz, 1H), 5.50 (t, J = 6.0 Hz, 1H), 4.52 (d, J = 6.0 Hz, 2H), 3.88-3.82 ( m,3H),3.18-3.09(m,2H),3.06(d,J=16.0Hz,1H), 2.63(d,J=15.5Hz,1H),1.92-1.86(m,1H),1.81-1.76 (m, 1H), 1.56-1.53 (m, 1H), 1.14-1.13 (m, 1H).
实施例67化合物67的制备：Example 67 Preparation of Compound 67:
Figure PCTCN2020072774-appb-000061
Figure PCTCN2020072774-appb-000061
步骤1：化合物67-1的制备Step 1: Preparation of compound 67-1
将500mg化合物M22-SM和603mg化合物M17溶解于9.0mL二氧六环中，加入655mg N,N-二异丙基乙胺。70℃搅拌反应2hrs。TLC检测反应完全，将反应液过滤，滤饼用DCM(10mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得453mg黄色固体化合物67-1。500 mg of compound M22-SM and 603 mg of compound M17 were dissolved in 9.0 mL of dioxane, and 655 mg of N,N-diisopropylethylamine was added. The reaction was stirred at 70°C for 2hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (10 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 453 mg of yellow solid compound 67-1.
步骤2：化合物67-2的制备Step 2: Preparation of compound 67-2
将453mg化合物67-1溶解于10mL的无水DCM中。在氮气保护下，-78℃缓慢滴加4.0mL 1.0M DIBAL-H的正己烷溶液，-78℃搅拌反应0.5hrs。LCMS检测反应完全，低温下依次滴加0.5mL水，0.5mL 2.5M NaOH水溶液淬灭，恢复室温再滴加0.5mL水，过滤后有机相无水硫酸钠干燥，减压浓缩得330mg粗品化合物67-2。453 mg of compound 67-1 was dissolved in 10 mL of anhydrous DCM. Under the protection of nitrogen, slowly add 4.0mL 1.0M DIBAL-H n-hexane solution dropwise at -78℃, and stir for 0.5hrs at -78℃. The reaction was completed by LCMS, 0.5 mL of water and 0.5 mL of 2.5M NaOH aqueous solution were added dropwise to quench at low temperature, and 0.5 mL of water was added dropwise after returning to room temperature. After filtration, the organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 330 mg of crude compound 67 -2.
步骤3：化合物67-3的制备Step 3: Preparation of compound 67-3
将330mg化合物67-2溶解于10mL的无水THF中。在氮气保护下，0℃加入60mg NaBH 4，恢复室温搅拌反应0.5hrs。LCMS检测反应完全，冰浴下滴加入5mL水淬灭，恢复室温，真空浓缩旋干THF后，水相用DCM(5mL×3)萃取，合并有机相无水硫酸钠干燥，减压浓缩。残余物经柱层析纯化得387mg黄色固体化合物67-3。 330 mg of compound 67-2 was dissolved in 10 mL of anhydrous THF. Under the protection of nitrogen, 60mg NaBH 4 was added at 0°C, and the reaction was stirred at room temperature for 0.5 hrs. LCMS detected that the reaction was complete, and quenched by adding 5 mL of water dropwise under ice bath, returned to room temperature, concentrated in vacuo and spin-dried THF, the aqueous phase was extracted with DCM (5 mL×3), the combined organic phase was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 387 mg of yellow solid compound 67-3.
步骤4：化合物67-4的制备Step 4: Preparation of compound 67-4
将200mg化合物67-3和99mg化合物22-3溶解于5mL二氧六环中，加入36mg Pd 2(dba) 3、45mg Xantphos和151mg N,N-二异丙基乙胺。氮气置换三次，氮气保护下反应升至70℃搅拌反应2hrs。LCMS和TLC检测反应完全，将反应液过滤，滤饼用DCM(15mL×2)洗涤，滤液减压浓缩，残余物经柱层析纯化得160mg化合物67-4。 200 mg of compound 67-3 and 99 mg of compound 22-3 were dissolved in 5 mL of dioxane, and 36 mg of Pd 2 (dba) 3 , 45 mg of Xantphos and 151 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 70°C and stirred for 2hrs. The reaction was completed by LCMS and TLC, the reaction solution was filtered, the filter cake was washed with DCM (15 mL×2), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 160 mg of compound 67-4.
步骤5：化合物67的制备Step 5: Preparation of compound 67
将50mg化合物67-4溶解于1.5mL二氧六环和0.3mL MeOH中，加入2N HCl(0.5mL，甲醇溶液)，RT搅拌反应0.5hrs。LCMS和TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(0.5mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(0.5mL)洗涤，固体再用DCM溶解，干燥后真空浓缩，经制备板纯化得13.3mg白色固体化合物67。 50 mg of compound 67-4 was dissolved in 1.5 mL of dioxane and 0.3 mL of MeOH, 2N HCl (0.5 mL, methanol solution) was added, and the reaction was stirred at RT for 0.5 hrs. LCMS and TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H 2 O (0.5 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (0.5 mL), dissolve the solid with DCM, dry and concentrate in vacuo , Purified by preparation plate to obtain 13.3 mg of white solid compound 67.
[M+H +]＝454.37。 [M+H + ]=454.37.
1H NMR(500MHz,DMSO-d 6)δ8.34(s,1H),7.96(d,J＝5.5Hz,1H),7.31(dd,J＝5.5,8.0Hz,1H),7.04-6.94(m,2H),6.73(s,2H),6.08(d,J＝5.5Hz,1H),5.51(t,J＝6.0Hz,1H),4.52(d,J＝5.5Hz,2H),3.89-3.78(m,3H),3.20-3.10(m,2H),3.05(d,J＝16.0Hz,1H),2.63(d,J＝16.0Hz,1H),1.90-1.85(m,1H),1.82-1.76(m,1H),1.56-1.53(m,1H),1.17-1.13(m,1H). 1 H NMR(500MHz,DMSO-d 6 )δ8.34(s,1H),7.96(d,J=5.5Hz,1H),7.31(dd,J=5.5,8.0Hz,1H),7.04-6.94( m, 2H), 6.73 (s, 2H), 6.08 (d, J = 5.5 Hz, 1H), 5.51 (t, J = 6.0 Hz, 1H), 4.52 (d, J = 5.5 Hz, 2H), 3.89- 3.78(m,3H), 3.20-3.10(m,2H), 3.05(d,J=16.0Hz,1H), 2.63(d,J=16.0Hz,1H),1.90-1.85(m,1H),1.82 -1.76 (m, 1H), 1.56-1.53 (m, 1H), 1.17-1.13 (m, 1H).
实施例68化合物68的制备：Example 68 Preparation of Compound 68:
Figure PCTCN2020072774-appb-000062
Figure PCTCN2020072774-appb-000062
步骤1：化合物68-1的制备Step 1: Preparation of compound 68-1
将500mg化合物68-0和418mg化合物M1-2溶解于10mL二氧六环中，加入95mg Pd 2(dba) 3、120mg Xantphos和900mg DIEA，氮气置换三次。升温至100℃反应16hrs。TLC检测反应完全，减压浓缩，残余物经柱层析纯化，得淡黄色固体0.7g化合物68-1。 500 mg of compound 68-0 and 418 mg of compound M1-2 were dissolved in 10 mL of dioxane, 95 mg of Pd 2 (dba) 3 , 120 mg of Xantphos, and 900 mg of DIEA were added, and nitrogen was replaced three times. The temperature was raised to 100°C for 16hrs. TLC detected that the reaction was complete, concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 0.7 g of compound 68-1 as a pale yellow solid.
步骤2：化合物68-2的制备Step 2: Preparation of compound 68-2
将0.7g化合物68-1溶解于3mL THF中，室温下加入0.66mg tBuOK，室温搅拌反应0.5hrs。TLC检测反应完全，反应液过滤，滤饼用THF(5mL×2)洗涤，得灰色固体380mg化合物68-2。Dissolve 0.7 g of compound 68-1 in 3 mL of THF, add 0.66 mg of tBuOK at room temperature, and stir for 0.5 hrs at room temperature. TLC detected that the reaction was complete, the reaction solution was filtered, and the filter cake was washed with THF (5 mL×2) to obtain 380 mg of compound 68-2 as a gray solid.
步骤3：化合物68-3的制备Step 3: Preparation of compound 68-3
将200mg化合物M22-2和123mg化合物68-2溶解于4mL二氧六环中，加入74mg Pd 2(dba) 3、23mg Xantphos和104mg DIEA，氮气置换三次。反应液升温至70℃反应3hrs。TLC检测反应完全，浓缩，残余物经制备薄层色谱法纯化得101mg白色化合物68-3。 200 mg of compound M22-2 and 123 mg of compound 68-2 were dissolved in 4 mL of dioxane, 74 mg of Pd 2 (dba) 3 , 23 mg of Xantphos, and 104 mg of DIEA were added, and replaced with nitrogen three times. The reaction solution was heated to 70°C for 3hrs. TLC detected that the reaction was complete, concentrated, and the residue was purified by preparative thin layer chromatography to obtain 101 mg of white compound 68-3.
步骤4：化合物68的制备Step 4: Preparation of compound 68
将101mg化合物68-3溶解于1.5mL二氧六环中和0.5mL MeOH中，加入2N HCl(632μL，甲醇溶液)，RT搅拌反应1hrs，TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(4mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(4mL)洗涤，取滤饼真空干燥得50.3mg米黄色固体化合物68。 101 mg of compound 68-3 was dissolved in 1.5 mL of dioxane and 0.5 mL of MeOH, 2N HCl (632 μL, methanol solution) was added, and the reaction was stirred at RT for 1 hrs. TLC detected that the reaction was complete, and the reaction solution was concentrated under reduced pressure. Add H 2 O (4 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (4 mL), take the filter cake and vacuum dry to obtain 50.3 mg of beige solid compound 68.
[M+H +]＝450.25。 [M+H + ]=450.25.
1H NMR(500MHz,CDCl3)δ8.36(s,1H),8.01(br,1H),7.34(d,J＝6.5Hz,1H),7.26-7.21(m,3H),6.24(d,J＝5.5Hz,1H),5.02-5.00(m,1H),4.70(s,2H),4.01(s,1H),3.64-3.59(m,2H),3.23-3.14(m,2H),3.10(d,J＝16.0Hz,1H),2.89(br,3H),2.74(d,J＝15.5Hz,1H),1.99-1.94(m,1H),1.90-1.84(m,1H),1.67-1.64(m,1H),1.43-1.40(m,1H). 1 H NMR(500MHz,CDCl3)δ8.36(s,1H),8.01(br,1H),7.34(d,J=6.5Hz,1H),7.26-7.21(m,3H),6.24(d,J =5.5Hz,1H),5.02-5.00(m,1H),4.70(s,2H),4.01(s,1H),3.64-3.59(m,2H),3.23-3.14(m,2H),3.10( d,J=16.0Hz,1H),2.89(br,3H),2.74(d,J=15.5Hz,1H),1.99-1.94(m,1H),1.90-1.84(m,1H),1.67-1.64 (m, 1H), 1.43-1.40 (m, 1H).
实施例69：化合物69的制备：Example 69: Preparation of Compound 69:
Figure PCTCN2020072774-appb-000063
Figure PCTCN2020072774-appb-000063
步骤1：化合物69-1的制备Step 1: Preparation of compound 69-1
N 2保护下，2.23g PPh 3溶于8mL二氧六环，加入1.15g NCS，室温搅拌0.5hrs，反应液变成浓稠的白色稀泥状，加入0.7g化合物M5，升温至100℃,搅拌1hrs。TLC检测反应完全,冷却至室温，7mL三乙胺，搅拌15mins，浓缩反应液,残余物经柱层析纯化，得 棕色油状物260mg化合物69-1。 Under N 2 protection, dissolve 2.23g PPh 3 in 8mL dioxane, add 1.15g NCS, stir at room temperature for 0.5hrs, the reaction solution will become thick white mud, add 0.7g compound M5, heat to 100℃, stir 1hrs. TLC detected that the reaction was complete, cooled to room temperature, 7 mL of triethylamine, stirred for 15 mins, concentrated the reaction solution, and the residue was purified by column chromatography to obtain 260 mg of compound 69-1 as a brown oil.
步骤2：化合物69-2的制备Step 2: Preparation of compound 69-2
将0.26g化合物69-1、342mg化合物M6和240mg DIPEA溶解于3mL二氧六环中，50℃搅拌反应2.5hrs。TLC检测反应完全，浓缩反应液,残余物经柱层析纯化，得白色固体280mg化合物69-2。0.26g of compound 69-1, 342mg of compound M6 and 240mg of DIPEA were dissolved in 3mL of dioxane, and the reaction was stirred at 50°C for 2.5hrs. TLC detected that the reaction was complete, the reaction solution was concentrated, and the residue was purified by column chromatography to obtain 280 mg of compound 69-2 as a white solid.
步骤3：化合物69-3的制备Step 3: Preparation of compound 69-3
将280mg化合物69-2溶解于3mL无水DCM中。在氮气保护下，-78℃缓慢滴加2.55mL 1.0M DIBAL-H的正己烷溶液，-78℃搅拌反应0.5hrs。LCMS检测反应完全，低温下依次滴加0.1mL水、0.1mL 2.5M NaOH水溶液淬灭，回室温再滴加0.25mL水，过滤后有机相无水硫酸钠干燥，减压浓缩得250mg粗品化合物69-3。280 mg of compound 69-2 was dissolved in 3 mL of anhydrous DCM. Under the protection of nitrogen, 2.55mL 1.0M DIBAL-H n-hexane solution was slowly added dropwise at -78℃, and the reaction was stirred at -78℃ for 0.5hrs. LCMS detects that the reaction is complete. Add 0.1 mL of water and 0.1 mL of 2.5M NaOH aqueous solution to quench at low temperature. Return to room temperature and then add 0.25 mL of water. After filtration, the organic phase is dried over anhydrous sodium sulfate and concentrated under reduced pressure to obtain 250 mg of crude compound 69 -3.
步骤3：化合物69-4的制备Step 3: Preparation of compound 69-4
将250mg化合物69-3溶解于5mL无水THF中。加入18mg NaBH 4，回室温搅拌反应0.5hrs。LCMS检测反应完全，滴加5mL水淬灭，然后加入DCM(5mL×2)萃取，合并有机相无水硫酸钠干燥，减压浓缩。残余物经柱层析纯化得180mg黄色固体化合物69-4。 250 mg of compound 69-3 was dissolved in 5 mL of anhydrous THF. 18mg NaBH 4 was added, and the reaction was stirred at room temperature for 0.5 hrs. LCMS detected that the reaction was complete, 5 mL of water was added dropwise to quench, then DCM (5 mL×2) was added for extraction, the combined organic phase was dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography to obtain 180 mg of yellow solid compound 69-4.
步骤4：化合物69-5的制备Step 4: Preparation of compound 69-5
将180mg化合物69-4和90mg化合物M1溶解于5mL二氧六环中，加入130mg Pd 2(dba) 3、41mg Xantphos和92mg N,N-二异丙基乙胺。氮气置换三次，氮气保护下反应升至70℃搅拌反应3hrs。LCMS和TLC检测反应完全，反应液减压浓缩，残余物经柱层析纯化得118mg化合物69-5。 180 mg of compound 69-4 and 90 mg of compound M1 were dissolved in 5 mL of dioxane, and 130 mg of Pd 2 (dba) 3 , 41 mg of Xantphos and 92 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 70°C and stirred for 3hrs. LCMS and TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 118 mg of compound 69-5.
步骤5：化合物69的制备Step 5: Preparation of compound 69
氮气保护下，将118mg化合物69-5溶解于1.5mL二氧六环中和0.5mL MeOH中，加入2N HCl(632μL，甲醇溶液)，室温搅拌反应1hrs，TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(4mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(4mL)洗涤，取滤饼真空干燥得63.2mg米黄色固体化合物69。 Under nitrogen protection, dissolve 118 mg of compound 69-5 in 1.5 mL of dioxane and 0.5 mL of MeOH, add 2N HCl (632 μL, methanol solution), stir and react at room temperature for 1 hrs, TLC detects that the reaction is complete, the reaction solution is concentrated under reduced pressure . Add H 2 O (4 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (4 mL), take the filter cake and vacuum dry to obtain 63.2 mg of beige solid compound 69.
[M+H+]＝450.29[M+H+]=450.29
1H NMR(500MHz,DMSO-d6)δ：7.94(d,J＝5.5Hz,1H),7.36-7.30(m,1H),7.20-7.15(m,3H),6.70(s,2H),5.98(d,J＝5.5Hz,1H),5.44(t,J＝6.0Hz,1H),4.48(d,J＝5.5Hz,2H),3.96-3.83(m,3H),3.21-3.02(m,2H),3.07(d,J＝15.5Hz,1H),2.64(d,J＝15.5Hz,1H),2.43(s,3H),1.92-1.87(m,1H),1.82-1.75(m,1H),1.56-1.53(m,1H),1.17-1.14(m,1H). 1 H NMR (500MHz, DMSO-d6) δ: 7.94 (d, J = 5.5 Hz, 1H), 7.36-7.30 (m, 1H), 7.20-7.15 (m, 3H), 6.70 (s, 2H), 5.98 (d,J=5.5Hz,1H), 5.44(t,J=6.0Hz,1H), 4.48(d,J=5.5Hz,2H),3.96-3.83(m,3H),3.21-3.02(m, 2H),3.07(d,J=15.5Hz,1H),2.64(d,J=15.5Hz,1H),2.43(s,3H),1.92-1.87(m,1H),1.82-1.75(m,1H ), 1.56-1.53 (m, 1H), 1.17-1.14 (m, 1H).
实施例70化合物70的制备：Example 70 Preparation of compound 70:
Figure PCTCN2020072774-appb-000064
Figure PCTCN2020072774-appb-000064
步骤1：化合物70-1的制备Step 1: Preparation of compound 70-1
将5.0g化合物M22-2溶解于50ml甲醇中，加入氨水(60ml，25％)，然后RT反应2.5小时。TLC检测反应完全，减压浓缩(把大部分甲醇旋掉)，再加入MeOH(10ml)搅拌5分钟，过滤，滤饼用MeOH(3ml)洗涤，滤饼烘干得4.75g固体即化合物70-1。Dissolve 5.0 g of compound M22-2 in 50 ml of methanol, add ammonia water (60 ml, 25%), and then react at RT for 2.5 hours. TLC detects the reaction is complete, concentrates under reduced pressure (revolve most of the methanol), add MeOH (10ml) and stir for 5 minutes, filter, wash the filter cake with MeOH (3ml), dry the filter cake to obtain 4.75g of solid compound 70- 1.
步骤2：化合物70-2的制备Step 2: Preparation of compound 70-2
将3.5g化合物70-1、3.82g化合物M6和5.04g DIPEA溶解于40mL THF中，55℃搅拌反应12hrs。TLC检测反应完全，反应液过滤,滤饼用DCM(40ml)洗涤，滤液减压浓缩，残余物经柱层析纯化，得白色固体5.66g化合物70-2。Dissolve 3.5 g of compound 70-1, 3.82 g of compound M6 and 5.04 g of DIPEA in 40 mL of THF, and stir at 55°C for 12 hrs. TLC detected that the reaction was complete, the reaction solution was filtered, the filter cake was washed with DCM (40 ml), the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 5.66 g of compound 70-2 as a white solid.
步骤3：化合物70-3的制备Step 3: Preparation of compound 70-3
将100mg化合物70-2和45mg化合物22-3溶解于2mL二氧六环中，加入18mg Pd 2(dba) 3、23mg Xantphos和51mg N,N-二异丙基乙胺。氮气置换三次，氮气保护下反应升至100℃搅拌反应3hrs。LCMS和TLC检测反应完全，反应液减压浓缩，残余物经柱层析纯化得42mg化合物70-3。 100 mg of compound 70-2 and 45 mg of compound 22-3 were dissolved in 2 mL of dioxane, and 18 mg of Pd 2 (dba) 3 , 23 mg of Xantphos and 51 mg of N,N-diisopropylethylamine were added. Replace with nitrogen for three times. Under the protection of nitrogen, the reaction was raised to 100°C and stirred for 3hrs. LCMS and TLC detected that the reaction was complete, the reaction solution was concentrated under reduced pressure, and the residue was purified by column chromatography to obtain 42 mg of compound 70-3.
步骤4：化合物70的制备Step 4: Preparation of compound 70
氮气保护下，将42mg化合物70-3溶解于0.5mL二氧六环中和0.5mL MeOH中，加入2N HCl(0.19ml，甲醇溶液)，RT搅拌反应1hrs，TLC检测反应完全，反应液减压浓缩。残余物加入H 2O(2mL)溶解，用饱和NaHCO 3调节溶液至pH＝8，固体析出，过滤，滤饼用H 2O(2mL)洗涤，取滤饼真空干燥得17.8mg淡黄色固体化合物70。 Under the protection of nitrogen, dissolve 42 mg of compound 70-3 in 0.5 mL of dioxane and 0.5 mL of MeOH, add 2N HCl (0.19 mL, methanol solution), stir at RT and react for 1 hrs. TLC detects that the reaction is complete and the reaction solution is decompressed concentrate. Add H 2 O (2 mL) to the residue to dissolve, adjust the solution to pH=8 with saturated NaHCO 3 , solid precipitate, filter, wash the filter cake with H 2 O (2 mL), take the filter cake and vacuum dry to obtain 17.8 mg of light yellow solid compound 70.
[M+H+]＝449.32[M+H+]=449.32
1H NMR(500MHz,DMSO-d 6)δ：8.34(s,1H),8.00(s,1H),7.96(d,J＝4.5Hz,1H),7.64(s,1H),7.31(d,J＝6.5Hz,1H),7.20-7.13(m,3H),6.70(s,2H),6.08(d,J＝4.5Hz,1H),4.02-3.97(m,2H),3.85(s,1H),3.29-3.21(m,2H),3.07(d,J＝15.5Hz,1H),2.65(d,J＝15.5Hz,1H),1.83-1.69(m,2H),1.53-1.50(m,1H),1.15-1.12(m,1H). 1 H NMR (500MHz, DMSO-d 6 ) δ: 8.34 (s, 1H), 8.00 (s, 1H), 7.96 (d, J = 4.5 Hz, 1H), 7.64 (s, 1H), 7.31 (d, J = 6.5Hz, 1H), 7.20-7.13 (m, 3H), 6.70 (s, 2H), 6.08 (d, J = 4.5 Hz, 1H), 4.02-3.97 (m, 2H), 3.85 (s, 1H) ), 3.29-3.21 (m, 2H), 3.07 (d, J = 15.5 Hz, 1H), 2.65 (d, J = 15.5 Hz, 1H), 1.83-1.69 (m, 2H), 1.53-1.50 (m, 1H), 1.15-1.12 (m, 1H).
对照例Control example
对照例1Comparative example 1
按照WO2018172984中的EXAMPLE 48所描述的方法，制备如下对照例1。According to the method described in EXAMPLE 48 in WO2018172984, the following Comparative Example 1 was prepared.
Figure PCTCN2020072774-appb-000065
Figure PCTCN2020072774-appb-000065
对照例2Comparative example 2
按照WO2019183367中的Example 243所描述的方法，制备如下对照例2。According to the method described in Example 243 in WO2019183367, the following Comparative Example 2 was prepared.
Figure PCTCN2020072774-appb-000066
Figure PCTCN2020072774-appb-000066
药理试验Pharmacological test
实施例A：SHP2变构抑制酶活测定Example A: SHP2 allosteric inhibition enzyme activity assay
SHP2通过双-酪氨酰-磷酰化的肽与其Src同源2(SH2)结构域的结合而变构活化。该在后的活化步骤导致SHP2的自动抑制界面的释放，这又使该SHP2蛋白酪氨酸磷酸酶(PTP)活化并可用于底物识别和反应催化。在迅速荧光测定版式中使用替代物DiFMUP监测SHP2的催化活性。SHP2 is allosterically activated by the binding of a bis-tyrosyl-phosphorylated peptide to its Src homology 2 (SH2) domain. The subsequent activation step results in the release of the SHP2 auto-inhibitory interface, which in turn activates the SHP2 protein tyrosine phosphatase (PTP) and can be used for substrate recognition and reaction catalysis. The surrogate DiFMUP was used to monitor the catalytic activity of SHP2 in the rapid fluorescence assay format.
试验步骤：experiment procedure:
(1)化合物配制：(1) Compound preparation:
用100％DMSO将本发明化合物(10mM储液)稀释成合适倍数，本发明化合物最终测试浓度为10μM、3.3333μM、1.1111μM、0.3704μM、0.1235μM、0.0412μM、0.0137μM、0.0046μM、0.0015μM、0.00μM；The compound of the present invention (10mM stock solution) was diluted to an appropriate multiple with 100% DMSO. The final test concentration of the compound of the present invention was 10μM, 3.3333μM, 1.1111μM, 0.3704μM, 0.1235μM, 0.0412μM, 0.0137μM, 0.0046μM, 0.0015μM , 0.00μM;
(2)准备酶反应工作液：(2) Prepare enzyme reaction working solution:
在室温下在96孔黑色聚苯乙烯板(平底、低凸缘、非结合表面)(Perki Elmer,Cat#6005270)中，使用50μL的最终反应体积和以下测定缓冲条件进行SHP2酶活检测：60mM HEPES，75mM NaCl，75mM KCl，0.05％BRIJ-35，1mM EDTA，5mMDTT。Perform SHP2 enzyme activity detection in a 96-well black polystyrene plate (flat bottom, low flange, non-binding surface) (Perki Elmer, Cat#6005270) at room temperature, using a final reaction volume of 50 μL and the following assay buffer conditions: 60mM HEPES, 75mM NaCl, 75mM KCl, 0.05% BRIJ-35, 1mM EDTA, 5mMDTT.
(3)酶催化反应及数据监测：(3) Enzyme catalytic reaction and data monitoring:
取本发明化合物加到对应的96孔板中，设置不加化合物和酶只加缓冲液的做为空白试 验孔。将SHP2Activating Peptide(IRS1_pY1172(dPEG8)pY1222)置于冰上融化，每孔加入25μM，然后取0.2ng SHP2蛋白样品加到对应孔板中，室温孵育1小时。加入替代底物DiFMUP(Invitrogen，Cat#D6567)加入反应，室温反应2小时后。采用分别使用340nm和450nm的激发波长和发射波长的酶标仪(Envision，Perki Elmer)监测荧光信号。Take the compound of the present invention and add it to the corresponding 96-well plate, and set the blank test well without adding the compound and enzyme and only adding buffer. Put SHP2Activating Peptide (IRS1_pY1172(dPEG8)pY1222) on ice to melt, add 25μM to each well, then add 0.2ng SHP2 protein sample to the corresponding well plate, and incubate at room temperature for 1 hour. The substitution substrate DiFMUP (Invitrogen, Cat#D6567) was added to the reaction, and the reaction was carried out at room temperature for 2 hours. The fluorescence signal was monitored by a microplate reader (Envision, Perki Elmer) with excitation wavelength and emission wavelength of 340nm and 450nm, respectively.
(4)数据分析：(4) Data analysis:
计算公式：Calculation formula:
抑制率％＝[1-(Conversion_ sample-Conversion_ min)/(Conversion_ max-Conversion_ min)]×100％ Inhibition rate%=[1-(Conversion_ sample -Conversion_ min )/(Conversion_ max -Conversion_ min )]×100%
其中：Conversion_sample是样品的转化率读数；Conversion_min是空白对照孔均值，代表没有酶活孔的转化率读数；Conversion_max是阳性对照孔比值均值，代表没有化合物抑制孔的转化率读数。采用分析软件GraphPad Prism的log(inhibitor)vs.response-Variable slope拟合量效曲线，并计算化合物对酶活性的IC 50值。 Among them: Conversion_sample is the conversion rate reading of the sample; Conversion_min is the average value of the blank control well, representing the conversion rate reading of the wells without enzyme activity; Conversion_max is the average value of the positive control well ratio, representing the conversion rate reading of the wells without compound inhibition. The analysis software GraphPad Prism log (inhibitor) vs. response-Variable slope was used to fit the dose-response curve, and the IC 50 value of the compound to the enzyme activity was calculated.
部分实施例的IC 50数据如表4所示。 The IC 50 data of some examples are shown in Table 4.
表4Table 4
化合物序号Compound number 化合物对SHP2酶活IC 50(nM) Compound to SHP2 enzyme activity IC 50 (nM)
化合物2Compound 2 8.568.56
化合物4Compound 4 3.193.19
化合物5Compound 5 12.6412.64
化合物7Compound 7 6.126.12
化合物8Compound 8 4.954.95
化合物9Compound 9 5.435.43
化合物10Compound 10 13.6413.64
化合物11Compound 11 13.8213.82
化合物12Compound 12 7.197.19
化合物13Compound 13 20.3720.37
化合物14Compound 14 20.6620.66
化合物15Compound 15 4.974.97
化合物16Compound 16 6.106.10
化合物17Compound 17 10.910.9
化合物18Compound 18 10.310.3
化合物19Compound 19 2.172.17
化合物20Compound 20 3.283.28
化合物21Compound 21 0.620.62
化合物22Compound 22 0.370.37
化合物23Compound 23 2.12.1
化合物24Compound 24 2.622.62
化合物25Compound 25 1.31.3
化合物26Compound 26 2.412.41
化合物27Compound 27 1.651.65
化合物29Compound 29 2.02.0
化合物30Compound 30 0.780.78
化合物31Compound 31 1.01.0
化合物32Compound 32 1.471.47
化合物33Compound 33 0.90.9
化合物34Compound 34 1.71.7
化合物35Compound 35 0.70.7
化合物36Compound 36 1.191.19
化合物37Compound 37 1.21.2
化合物38Compound 38 0.60.6
化合物39Compound 39 0.710.71
化合物40Compound 40 1.801.80
化合物41Compound 41 0.810.81
化合物42Compound 42 0.850.85
化合物43Compound 43 2.412.41
化合物44Compound 44 0.830.83
化合物45Compound 45 1.601.60
化合物46Compound 46 1.101.10
化合物47Compound 47 0.600.60
化合物48Compound 48 1.71.7
化合物49Compound 49 4.14.1
化合物50Compound 50 0.70.7
化合物51Compound 51 //
化合物52Compound 52 //
化合物53Compound 53 22
化合物54Compound 54 11
化合物55Compound 55 9.39.3
化合物56Compound 56 6.86.8
化合物57Compound 57 //
化合物58Compound 58 0.720.72
化合物59Compound 59 2.152.15
化合物60Compound 60 7.197.19
化合物61Compound 61 19.7519.75
化合物62Compound 62 1.41.4
化合物63Compound 63 4.34.3
化合物64Compound 64 2.02.0
化合物65Compound 65 2.12.1
化合物66Compound 66 3.13.1
化合物67Compound 67 2.02.0
化合物68Compound 68 1.81.8
化合物69Compound 69 4.74.7
化合物70Compound 70 2.32.3
注：“/”表示未测试。Note: "/" means not tested.
本发明的化合物对SHP2蛋白具有变构抑制作用。The compound of the present invention has an allosteric inhibitory effect on SHP2 protein.
实施例B：细胞增殖试验Example B: Cell Proliferation Test
使用体外细胞试验评估本发明的化合物对肺鳞癌细胞KYSE-520细胞和白血病细胞MV-4-11细胞增殖的影响。试验中所用的检测方法是CELL TITER-GLO(CTG)发光法， 该法可通过对ATP进行定量测定来检测活细胞数目。因为ATP参与生物体内多种酶促反应，是活细胞新陈代谢的一个指标，其含量直接反应了细胞的数量及细胞状态，实验过程中向细胞培养基加入CellTiter-Glo TM试剂，测量发光值，发光值与ATP量成正比，而ATP又和活细胞数正相关，因此可通过检测ATP含量考察细胞活力。 An in vitro cell assay was used to evaluate the effects of the compounds of the present invention on the proliferation of lung squamous cell carcinoma KYSE-520 cells and leukemia cells MV-4-11 cells. The detection method used in the experiment is the CELL TITER-GLO (CTG) luminescence method, which can detect the number of living cells by quantitatively measuring ATP. Because ATP participates in a variety of enzymatic reactions in organisms, it is an indicator of living cell metabolism. Its content directly reflects the number and cell state of cells. During the experiment, CellTiter-Glo TM reagent was added to the cell culture medium to measure the luminescence value. The value is directly proportional to the amount of ATP, and ATP is positively related to the number of living cells, so cell viability can be inspected by detecting ATP content.
试验步骤：experiment procedure:
(1)细胞铺板：(1) Cell plating:
取一瓶对数生长期的KYSE-520细胞，消化重悬细胞后计数，调整细胞密度后接种到96孔板中，每孔接种1000个细胞，孔板置于37℃、5％CO 2的培养箱中培养24hrs后加入本发明化合物进行处理； Take a bottle of KYSE-520 cells in logarithmic growth phase, digest the resuspended cells, count them, adjust the cell density and inoculate them into a 96-well plate, inoculate 1000 cells per well, and place the well plate at 37℃, 5% CO 2 After culturing in an incubator for 24hrs, adding the compound of the present invention for treatment;
取一瓶对数生长期的MV-4-11细胞，消化重悬细胞后计数，调整细胞密度后接种到96孔板中，每孔接种4000个细胞，孔板置于37℃、5％CO 2的培养箱中培养24hrs后加入本发明化合物进行处理； Take a bottle of MV-4-11 cells in logarithmic growth phase, digest and resuspend the cells, count them, adjust the cell density and inoculate them in a 96-well plate, inoculate 4000 cells per well, and place the plate at 37°C, 5% CO 2. After culturing in the incubator for 24hrs, adding the compound of the present invention for treatment;
(2)细胞化合物处理：(2) Cell compound treatment:
配取适量本发明化合物进行细胞处理，化合物终浓度从高至低依次为1000nM、333.3nM、111.1nM、37.04nM、12.35nM、4.115nM、1.372nM、0.4572nM、0.1524nM、0nM，孔板放入37℃,5％CO 2培养箱培养120hrs。只加培养基不加细胞孔设为调零组；化合物浓度为0nM组为空白组。 Prepare an appropriate amount of the compound of the present invention for cell treatment, and the final concentration of the compound from high to low is 1000nM, 333.3nM, 111.1nM, 37.04nM, 12.35nM, 4.115nM, 1.372nM, 0.4572nM, 0.1524nM, 0nM. Enter 37°C, 5% CO 2 incubator for 120hrs. Only adding medium without adding cell wells is set as the zero adjustment group; the compound concentration of 0 nM group is the blank group.
(3)CTG检测：(3) CTG detection:
细胞培养120hrs后每孔加入50μL的
Figure PCTCN2020072774-appb-000067
Luminescent Cell Viability Assay溶液，轻轻震荡2mins，室温继续孵育10mins，在多功能酶标仪上读取各孔的检测数值。 After cell culture for 120hrs, add 50μL to each well
Figure PCTCN2020072774-appb-000067
Luminescent Cell Viability Assay solution, gently shake for 2mins, continue to incubate at room temperature for 10mins, and read the detection value of each well on the multifunctional microplate reader.
(4)数据分析：(4) Data analysis:
根据发光值读数计算抑制率，Calculate the inhibition rate based on the luminous value reading,
抑制率％＝(1-(给药组值-调零组值)/(空白组值-调零组值)*100Inhibition rate%=(1-(administration group value-zero adjustment group value)/(blank group value-zero adjustment group value)*100
GraphPad Prism的log(inhibitor)vs.response-Variable slope拟合量效曲线并计算化合物抑制细胞增殖的IC 50。 GraphPad Prism the log (inhibitor) vs.response-Variable slope fitting dose-response curve and calculate IC 50 of compounds to inhibit cell proliferation.
实验数据如表5所示。The experimental data is shown in Table 5.
表5table 5
Figure PCTCN2020072774-appb-000068
Figure PCTCN2020072774-appb-000068
Figure PCTCN2020072774-appb-000069
Figure PCTCN2020072774-appb-000069
Figure PCTCN2020072774-appb-000070
Figure PCTCN2020072774-appb-000070
注：“/”表示未测试。Note: "/" means not tested.
本发明的化合物对KYSE-520细胞的增殖和MV-4-11细胞的增殖具有良好的抑制作用。The compound of the present invention has a good inhibitory effect on the proliferation of KYSE-520 cells and the proliferation of MV-4-11 cells.
实施例C：hERG钾离子通道的抑制试验Example C: Inhibition test of hERG potassium ion channel
采用全细胞膜片钳技术检测待测化合物对hERG通道的阻断作用。The whole cell patch clamp technique was used to detect the blocking effect of the test compound on the hERG channel.
细胞培养Cell culture
hERG钾通道稳定表达的HEK293细胞系在含有10％胎牛血清及0.8mg/mL G418的DMEM培养基中培养，培养温度为37℃，二氧化碳浓度为5％。The HEK293 cell line stably expressing the hERG potassium channel was cultured in DMEM medium containing 10% fetal bovine serum and 0.8 mg/mL G418 at a culture temperature of 37°C and a carbon dioxide concentration of 5%.
细胞传代：除去旧培养基并用PBS洗一次，然后加入1mL TrypLE TM Express溶液，37℃孵育0.5分钟。当细胞从皿底脱离，加入5mL 37℃预热的完全培养基。将细胞悬液用吸管轻轻吹打使聚集的细胞分离。将细胞悬液转移至无菌的离心管中，1000rpm离心5分钟收集细胞。扩增或维持培养，将细胞接种于6厘米细胞培养皿，每个细胞培养皿，接种细胞量为2.5*105cells(最终体积：5mL)。 Cell passage: Remove the old medium and wash once with PBS, then add 1mL TrypLE TM Express solution and incubate at 37°C for 0.5 minutes. When the cells detach from the bottom of the dish, add 5 mL of complete medium pre-warmed at 37°C. The cell suspension was gently pipetted to separate the aggregated cells. The cell suspension was transferred to a sterile centrifuge tube and centrifuged at 1000 rpm for 5 minutes to collect the cells. To expand or maintain the culture, inoculate the cells in 6 cm cell culture dishes, each cell culture dish, the amount of cells inoculated is 2.5*105 cells (final volume: 5mL).
为维持细胞的电生理活性，细胞密度必须不能超过80％。In order to maintain the electrophysiological activity of cells, the cell density must not exceed 80%.
膜片钳检测，实验之前细胞用TrypLE TM Express分离，将3*103细胞铺到盖玻片上，在24孔板中培养(最终体积：500μL)，18个小时后，进行实验检测。 For patch clamp detection, the cells were separated with TrypLE TM Express before the experiment, and 3*103 cells were spread on a cover glass and cultured in a 24-well plate (final volume: 500 μL). After 18 hours, the experiment was performed.
细胞内外液Intracellular fluid
细胞外液:140mM NaCl,3.5mM KCl,1mM MgCl 2·6H 2O,2mM CaCl 2,10mM D-葡萄糖,10mM HEPES,1.25mM NaH 2PO 4,NaOH调节pH＝7.4。 Extracellular fluid: 140mM NaCl, 3.5mM KCl, 1mM MgCl 2 ·6H 2 O, 2mM CaCl 2 , 10mM D-glucose, 10mM HEPES, 1.25mM NaH 2 PO 4 , NaOH to adjust pH=7.4.
细胞内液:20mM KCl,115mM K-Aspartic,1mM MgCl 2·6H 2O,5mM EGTA,10mM HEPES,2mM Na 2-ATP,KOH调节pH＝7.2。 Intracellular fluid: 20mM KCl, 115mM K-Aspartic, 1mM MgCl 2 ·6H 2 O, 5mM EGTA, 10mM HEPES, 2mM Na 2 -ATP, KOH to adjust pH=7.2.
化合物的配制Compound formulation
用细胞外液将被测化合物储液进行稀释，配成10μΜ工作液，或者梯度稀释为0.3μΜ,1μΜ,3μΜ,10μM,30μM溶液。Dilute the test compound stock solution with extracellular fluid to prepare a 10 μM working solution, or a gradient dilution to a solution of 0.3 μM, 1 μM, 3 μM, 10 μM, 30 μM.
目测被测化合物的溶解性，被测化合物全部溶解没有肉眼可见的沉淀。The solubility of the test compound was visually checked, and the test compound was completely dissolved without visible precipitation.
西沙必利(阳性对照)Cisapride (positive control)
将称量出的1.2mg西沙必利用243μL DMSO配制成10mM的储液。The weighed 1.2mg cisap must be prepared into a 10mM stock solution using 243μL DMSO.
将西沙必利储液用DMSO依次以10倍的稀释倍数由高到低稀释至10μM的稀释液。The cisapride stock solution was diluted successively with DMSO at a 10-fold dilution from high to low to a 10μM dilution.
用细胞外液将10μM西沙必利稀释液进行稀释，配成10nΜ工作液。Dilute the 10 μM cisapride diluent with extracellular fluid to prepare a 10 nM working solution.
目测西沙必利的溶解性，西沙必利全部溶解没有肉眼可见的沉淀。Visually inspecting the solubility of cisapride, cisapride was completely dissolved without visible precipitation.
实验方法参照：Experimental method reference:
全细胞膜片钳记录全细胞hERG钾电流的电压刺激方案如下：当形成全细胞封接后细胞膜电压钳制于-80mV。钳制电压由-80mV除极至-50mV维持0.5秒，然后阶跃至30mV维持2.5秒，再迅速恢复至-50mV维持4秒可以激发出hERG通道的尾电流。每隔10秒重复采集数据，观察药物对hERG尾电流的作用。以0.5秒的-50mV刺激为漏电流检测。实验数据由EPC-10放大器(HEKA)进行采集并储存于PatchMaster(HEKA)软件中。The whole cell patch clamp voltage stimulation scheme for recording the whole cell hERG potassium current is as follows: when the whole cell seal is formed, the cell membrane voltage is clamped to -80mV. The clamping voltage is depolarized from -80mV to -50mV for 0.5 seconds, then stepped to 30mV for 2.5 seconds, and then quickly restored to -50mV for 4 seconds to stimulate the tail current of the hERG channel. Collect data repeatedly every 10 seconds to observe the effect of the drug on the hERG tail current. A stimulus of -50mV for 0.5 seconds was used as leakage current detection. The experimental data is collected by EPC-10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.
用微电极拉制仪将毛细玻璃管拉制成记录电极。在倒置显微镜下操纵微电极操纵仪将记录电极接触到细胞上，给予负压抽吸，形成GΩ封接。形成GΩ封接后进行快速电容补偿，然后继续给予负压，吸破细胞膜，形成全细胞记录模式。然后进行慢速电容的补偿并记录膜电容及串联电阻。不给予漏电补偿。The capillary glass tube is drawn into a recording electrode with a microelectrode drawing instrument. Operate the microelectrode manipulator under the inverted microscope to contact the recording electrode on the cell, and apply negative pressure suction to form a GΩ seal. After forming the GΩ seal, perform fast capacitance compensation, and then continue to give negative pressure to suck and break the cell membrane to form a whole-cell recording mode. Then perform slow capacitance compensation and record the film capacitance and series resistance. No leakage compensation is given.
当全细胞记录的hERG电流稳定后开始给药，每个药物浓度作用至5分钟(或者电流至稳定)。将铺有细胞的盖玻片置于倒置显微中的记录浴槽中，测试化合物以及不含化合物的外液利用重力灌流的方法依次流经记录浴槽从而作用于细胞，在记录中利用真空泵进行液体交换。每一个细胞在不含化合物的外液中检测到的电流作为自己的对照组。独立重复检测多个细胞。所有电生理实验在室温下进行。When the hERG current recorded by the whole cell stabilizes, the drug is administered, and each drug concentration acts for 5 minutes (or the current stabilizes). Place the cover glass covered with cells in the recording bath in the inverted microscope. The test compound and the compound-free external fluid flow through the recording bath sequentially by gravity perfusion to act on the cells, and the vacuum pump is used to carry out the liquid during recording. exchange. The current detected by each cell in the compound-free external fluid serves as its own control group. Test multiple cells independently and repeatedly. All electrophysiological experiments were performed at room temperature.
数据质量标准Data quality standards
以下标准用来判断数据是否可以接受：The following criteria are used to determine whether the data is acceptable:
(1)串联电阻≤20MΩ(1) Series resistance ≤20MΩ
(2)封接电阻≥1GΩ(2) Sealing resistance≥1GΩ
(3)起始尾电流峰值≥400pA(3) Peak tail current peak ≥400pA
(4)起始尾电流峰值大于激活电流峰值(4) The initial tail current peak is greater than the activation current peak
(5)尾电流没有明显的自发性衰减(5分钟内自发性衰减小于5％)(5) There is no obvious spontaneous attenuation of tail current (the spontaneous attenuation is less than 5% within 5 minutes)
(6)在膜电位为-80mV下无明显的漏电流(漏电流≤100pA)(6) There is no obvious leakage current under the membrane potential of -80mV (leakage current ≤100pA)
数据分析data analysis
首先将每一个药物浓度作用后的电流和空白对照电流标准化
Figure PCTCN2020072774-appb-000071
然后计算每一个药物浓度对应的抑制率
Figure PCTCN2020072774-appb-000072
First, the current after each drug concentration is normalized to the blank control current
Figure PCTCN2020072774-appb-000071
Then calculate the inhibition rate corresponding to each drug concentration
Figure PCTCN2020072774-appb-000072
实施例化合物hERG测试结果见表6。See Table 6 for the hERG test results of the example compounds.
表6Table 6
编号Numbering 抑制率@10μMInhibition rate @10μM
对照例1Comparative example 1 95.12％95.12%
对照例2Comparative example 2 >95.6％>95.6%
化合物3Compound 3 86.7％86.7%
化合物5Compound 5 60.8％60.8%
化合物8Compound 8 97.36％97.36%
化合物9Compound 9 93.35％93.35%
化合物16Compound 16 87.99％87.99%
化合物18Compound 18 42.63％42.63%
化合物21Compound 21 82.3％82.3%
化合物22Compound 22 43.5％43.5%
化合物24Compound 24 79.9％79.9%
化合物26Compound 26 >90％>90%
化合物30Compound 30 92.2％92.2%
化合物33Compound 33 >90％>90%
化合物39Compound 39 83.7％83.7%
化合物40Compound 40 98.1％98.1%
化合物41Compound 41 85.6％85.6%
化合物42Compound 42 87.6％87.6%
化合物43Compound 43 56.6％56.6%
化合物44Compound 44 80.2％80.2%
化合物45Compound 45 78.0％78.0%
化合物46Compound 46 87.8％87.8%
化合物58Compound 58 69.4％69.4%
化合物59Compound 59 57.0％57.0%
化合物62Compound 62 >90％>90%
化合物64Compound 64 5.02％5.02%
化合物65Compound 65 62.1％62.1%
化合物66Compound 66 37.1％37.1%
化合物67Compound 67 29.5％29.5%
化合物68Compound 68 54.4％54.4%
化合物69Compound 69 26.3％26.3%
化合物70Compound 70 9.12％9.12%
我们出乎意料地发现，本发明的化合物在普遍对SHP2有较好的抑制活性的同时，当满足以下两个条件之一时，即1)、通式I中吡嗪环上R 1位置为羟甲基且A 1、A 3位置均为N，例如化合物22、化合物64～69；2)、通式I中吡嗪环上R 1位置为酰胺基且A 3位置为N，例如化合物5、化合物18、化合物58或化合物59，相比其他技术方案，可显著改善hERG抑制问题，有望降低心脏毒性。 We unexpectedly found that while the compounds of the present invention generally have good inhibitory activity on SHP2, when one of the following two conditions is met, namely 1), the position of R 1 on the pyrazine ring in formula I is a hydroxyl group. A methyl group and both A 1 and A 3 positions are N, such as compound 22, compounds 64-69; 2), in formula I, the R 1 position on the pyrazine ring is an amide group and A 3 position is N, such as compound 5, Compared with other technical solutions, compound 18, compound 58 or compound 59 can significantly improve the hERG inhibition problem and is expected to reduce cardiotoxicity.
实施例D：血浆蛋白结合测定Example D: Plasma protein binding assay
按照以下步骤测定血浆蛋白结合。Follow the steps below to determine plasma protein binding.
1)100mM磷酸钠和150mM NaCl缓冲液(PBS)的制备1) Preparation of 100mM sodium phosphate and 150mM NaCl buffer (PBS)
通过在去离子水中溶解14.2g/L Na 2HPO 4和8.77g/L NaCl来制备碱性溶液，该溶液可以在4℃下保存长达7天。通过将12.0g/L NaH 2PO 4和8.77g/L NaCl溶解在酸性溶液中可以制备酸性溶液，该溶液可在4℃下保存7天。将碱性溶液用酸性溶液滴定至pH 7.4，并在4℃下保存7天。在实验当天进行检查，如果pH超出规格7.4±0.1，则进行调整。 An alkaline solution was prepared by dissolving 14.2 g/L Na 2 HPO 4 and 8.77 g/L NaCl in deionized water, which can be stored at 4° C. for up to 7 days. An acidic solution can be prepared by dissolving 12.0g/L NaH 2 PO 4 and 8.77g/L NaCl in an acidic solution, which can be stored at 4°C for 7 days. The alkaline solution was titrated to pH 7.4 with an acidic solution and stored at 4°C for 7 days. Check on the day of the experiment and adjust if the pH exceeds the specification of 7.4±0.1.
2)血浆的制备2) Preparation of plasma
冷冻的血浆立即在室温下解冻。The frozen plasma was immediately thawed at room temperature.
将血浆以3,220g离心10分钟以去除凝块，并将上清液收集到新的试管中。检查并记录血浆的pH值。The plasma was centrifuged at 3,220g for 10 minutes to remove clots, and the supernatant was collected in a new test tube. Check and record the pH of the plasma.
注意：a).仅使用冻融化不超过两次的血浆。b).仅使用pH 7到pH 8范围内的血浆。Note: a). Only use plasma that has been frozen and thawed not more than twice. b). Only use plasma within the range of pH 7 to pH 8.
3)工作溶液的制备3) Preparation of working solution
用DMSO以400μM的浓度制备测试化合物和对照化合物酮康唑的工作溶液。然后移去4μL的工作溶液使与796μL的人、犬、猴、大鼠或小鼠血浆混合，最终得到浓度为2μM(0.5％DMSO)的混合溶液。将血浆样品彻底涡旋。Working solutions of the test compound and the control compound ketoconazole were prepared with DMSO at a concentration of 400 μM. Then 4 μL of the working solution was removed and mixed with 796 μL of human, dog, monkey, rat, or mouse plasma to finally obtain a mixed solution with a concentration of 2 μM (0.5% DMSO). Vortex the plasma sample thoroughly.
4)快速平衡透析步骤4) Quickly balance the dialysis steps
在红圈侧加入200μL血浆样品，在对侧加入400μL的透析缓冲液(PBS)透析。一式三份进行测定。将透析板密封并在37℃孵育箱中，于150rpm下孵育4小时。孵育结束时，移去密封，并将来自缓冲液室和血浆室的50μL样品转移至96孔板的孔中。Add 200 μL of plasma sample on the red circle side, and add 400 μL of dialysis buffer (PBS) on the opposite side for dialysis. The determination is performed in triplicate. The dialysis plate was sealed and incubated in a 37°C incubator at 150 rpm for 4 hours. At the end of the incubation, the seal was removed, and 50 μL of samples from the buffer chamber and the plasma chamber were transferred to the wells of the 96-well plate.
5)样品分析步骤5) Sample analysis steps
向每个缓冲液样品中加入50μL空白血浆，并向收集的血浆样品中补充等体积的PBS。加入300μL室温淬灭溶液(含有内标的乙腈(IS，10ng/mL拉贝洛尔和10ng/mL格列本脲))使蛋白质沉淀。将板中的样品涡旋5分钟，并在4℃下以3220g离心30分钟。然后用100μL或200μL水将100μL的上清液转移至新的96孔板中用于LC-MS/MS分析(取决于LC-MS信号响应和峰形)。Add 50 μL of blank plasma to each buffer sample, and add an equal volume of PBS to the collected plasma sample. 300 μL of room temperature quenching solution (acetonitrile containing internal standard (IS, 10 ng/mL labetalol and 10 ng/mL glibenclamide)) was added to precipitate the protein. The samples in the plate were vortexed for 5 minutes and centrifuged at 3220 g for 30 minutes at 4°C. Then transfer 100 μL of supernatant with 100 μL or 200 μL of water to a new 96-well plate for LC-MS/MS analysis (depending on LC-MS signal response and peak shape).
计算测试化合物和对照化合物结合的百分比，如下所示：Calculate the percentage of test compound and control compound binding as follows:
％游离＝(峰面积比 缓冲液室/峰面积比 血浆室)*100 % Free = (peak area ratio buffer chamber /peak area ratio plasma chamber )*100
％结合＝100-％游离% Binding = 100-% free
％回收＝(峰面积比 缓冲液室+峰面积比 血浆室)/峰面积比 总样品*100 % Recovery = (peak area ratio buffer chamber + peak area ratio plasma chamber )/peak area ratio total sample * 100
峰面积比 缓冲液室表示游离部分的浓度 Peak area ratio buffer chamber indicates the concentration of free part
峰面积比 血浆室表示游离和结合部分的浓度 Peak area ratio plasma chamber indicates the concentration of free and bound fraction
峰面积比 总样品表示培养前开始样品的浓度 The peak area ratio to the total sample indicates the concentration of the sample started before incubation
表7显示了不同物种中对照化合物和测试化合物的血浆蛋白结合结果。Table 7 shows the plasma protein binding results of the control compound and the test compound in different species.
表7Table 7
Figure PCTCN2020072774-appb-000073
Figure PCTCN2020072774-appb-000073
Figure PCTCN2020072774-appb-000074
Figure PCTCN2020072774-appb-000074
通常，只有未结合的部分才具有生物学作用或被代谢。因此，与血浆蛋白的结合程度会显著影响药物的药代动力学和药效学性质。Usually, only the unbound part has a biological effect or is metabolized. Therefore, the degree of binding to plasma proteins will significantly affect the pharmacokinetics and pharmacodynamic properties of the drug.
如表7所示，对比例反映了与血浆蛋白的高度结合，因此该药的功效可能降低。出乎意料的是，与对比例相比，本发明的示例性化合物具有较低的血浆蛋白结合度。预示本发明对人体具有优良的药代动力学和药效学性质。As shown in Table 7, the comparative example reflects a high degree of binding to plasma proteins, so the efficacy of the drug may be reduced. Unexpectedly, the exemplary compound of the present invention has a lower degree of plasma protein binding compared with the comparative example. It is predicted that the present invention has excellent pharmacokinetics and pharmacodynamic properties on the human body.
虽然本发明已通过其实施方式进行了全面的描述，但是值得注意的是，各种变化和修改对于本领域技术人员都是显而易见的。这样的变化和修改都应该包括在本发明所附权利要求的范围内。Although the present invention has been fully described through its embodiments, it is worth noting that various changes and modifications are obvious to those skilled in the art. Such changes and modifications should be included in the scope of the appended claims of the present invention.

Claims (52)

  1. 式I所示的化合物，或其药学上可接受的盐、互变异构体、溶剂化物、螯合物、非共价复合物或前体药物，The compound represented by formula I, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof,
    Figure PCTCN2020072774-appb-100001
    Figure PCTCN2020072774-appb-100001
    其中，among them,
    R 1任意地选自氨基、-C(O)-R a、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基； Optionally R 1 is selected from amino, -C (O) -R a, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, C 1-8 alkoxy containing substituents;
    R a任意地选自氨基、-NH-OH、C 1-3烷基； R a is optionally selected from amino, -NH-OH, C 1-3 alkyl;
    R 2任意地选自氢、C 1-4烷基或含取代基的C 1-4烷基； Optionally R 2 is selected from hydrogen, C 1-4 alkyl group or a substituted C 1-4 alkyl;
    R 3任意地选自氢、卤素、氨基、-C(O)NH 2、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基或含取代基的C 1-8烷氧基； Optionally R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
    R 4任意地选自氢、卤素、氨基、酰胺基、-C≡N、羧基、羟基、羟甲基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基或含取代基的C 2-8炔基； Optionally R 4 is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, hydroxymethyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl base;
    A 1任意地选自CR 5或N； A 1 is arbitrarily selected from CR 5 or N;
    A 2任意地选自CR 6或N； A 2 is arbitrarily selected from CR 6 or N;
    A 3任意地选自CR 7或N； A 3 is arbitrarily selected from CR 7 or N;
    A 4任意地选自CR 8或N； A 4 is arbitrarily selected from CR 8 or N;
    U任意地选自C(R 9) 2、O或NR 10； U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
    其中，R 5、R 6、R 7、R 8、R 9或R 10独立地选自氢、羟基、卤素、氨基、含取代基的氨基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基、含取代基的C 2-8炔基或C 5-6杂环基；或者， Wherein, R 5 , R 6 , R 7 , R 8 , R 9 or R 10 are independently selected from hydrogen, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted C 1 -8 alkyl, C 1-8 alkoxy, substituted C 1-8 alkoxy, C 2-8 alkenyl, substituted C 2-8 alkenyl, C 2-8 alkynyl, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents; or,
    R 5和R 6与他们连接的碳原子共同形成5到6元芳基或5到6元杂环基； R 5 and R 6 and the carbon atom to which they are attached together form a 5- to 6-membered aryl group or a 5- to 6-membered heterocyclic group;
    L选自S；L is selected from S;
    环A任意地选自C 6-10芳基或C 5-10杂芳基，所述C 5-10杂芳基含有一个或两个N或S杂原子； Ring A is optionally selected from a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group contains one or two N or S heteroatoms;
    Rx任意地选自氢、羟基、卤素、氰基、氨基、含取代基的氨基、磺酰基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 3-8环烷基或含取代基的C 3-8环烷基； Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;
    n为0、1、2、3或4。n is 0, 1, 2, 3, or 4.
  2. 根据权利要求1所述的化合物，其特征在于，R 1任意地选自氨基、-C(O)NH 2、-C≡N、C 1-3烷基、含取代基的C 1-3烷基或C 1-3烷氧基。 The compound according to claim 1, wherein, R & lt an arbitrarily selected from amino, -C (O) NH 2, -C≡N, C 1-3 alkyl group, a substituted group containing C 1-3 alkoxy Group or C 1-3 alkoxy.
  3. 根据权利要求1或2所述的化合物，其特征在于，R 1任意地选自氨基、-C(O)NH 2、-C≡N、羟基取代的甲基、
    Figure PCTCN2020072774-appb-100002
    The compound according to claim 1 or 2, wherein R 1 is arbitrarily selected from amino group, -C(O)NH 2 , -C≡N, hydroxy-substituted methyl,
    Figure PCTCN2020072774-appb-100002
  4. 根据权利要求1-3任一项所述的化合物，其特征在于，R 2为氢。 The compound of any one of claims 1-3, wherein R 2 is hydrogen.
  5. 根据权利要求1-4任一项所述的化合物，其特征在于，R 3任意地选自氢、卤素、C 1-3烷基或含取代基的C 1-3烷基。 A compound according to any one of claims 1-4, wherein, R 3 is optionally selected from hydrogen, halo, C 1-3 alkyl or an optionally substituted C 1-3 alkyl.
  6. 根据权利要求1-5任一项所述的化合物，其特征在于，R 3任意地选自氢、氯或甲基。 The compound according to any one of claims 1-5, wherein R 3 is arbitrarily selected from hydrogen, chlorine or methyl.
  7. 根据权利要求1-6任一项所述的化合物，其特征在于，R 4任意地选自氢、卤素、含取代基的C 1-8烷基、C 1-8烷氧基。 The compound according to any one of claims 1-6, wherein R 4 is arbitrarily selected from hydrogen, halogen, C 1-8 alkyl group containing substituents, and C 1-8 alkoxy group.
  8. 根据权利要求1-7任一项所述的化合物，其特征在于，R 4任意地选自氢、F、Cl、-CHF 2、CF 3或-O-CH 3。 The compound according to any one of claims 1-7, wherein R 4 is arbitrarily selected from hydrogen, F, Cl, -CHF 2 , CF 3 or -O-CH 3 .
  9. 根据权利要求1-8任一项所述的化合物，其特征在于，A 1为CR 5或N，其中R 5选自氢、卤素或卤素取代的C 1-3烷基。 The compound according to any one of claims 1-8, wherein A 1 is CR 5 or N, wherein R 5 is selected from hydrogen, halogen, or halogen substituted C 1-3 alkyl.
  10. 根据权利要求1-9任一项所述的化合物，其特征在于，A 1为CR 5或N，其中R 5选自氢、Cl或三氟甲基。 The compound according to any one of claims 1-9, wherein A 1 is CR 5 or N, wherein R 5 is selected from hydrogen, Cl or trifluoromethyl.
  11. 根据权利要求1-10任一项所述的化合物，其特征在于，A 2为CR 6或N，其中R 6选自氢、羟基、卤素、氨基或C 1-8烷氧基。 The compound according to any one of claims 1-10, wherein A 2 is CR 6 or N, wherein R 6 is selected from hydrogen, hydroxyl, halogen, amino or C 1-8 alkoxy.
  12. 根据权利要求1-11任一项所述的化合物，其特征在于，A 2为CR 6或N，其中R 6选自氢、OH、F、Cl、氨基或-O-CH 3。 The compound according to any one of claims 1-11, wherein A 2 is CR 6 or N, wherein R 6 is selected from hydrogen, OH, F, Cl, amino, or -O-CH 3 .
  13. 根据权利要求1-12任一项所述的化合物，其特征在于，A 3为CR 7或N，其中R 7为氢或卤素。 The compound according to any one of claims 1-12, wherein A 3 is CR 7 or N, wherein R 7 is hydrogen or halogen.
  14. 根据权利要求1-13任一项所述的化合物，其特征在于，A 4为CR 8或N，其中R 8选自氢、卤素、氨基、取代氨基、C 1-3烷氧基或C 5-6杂环基。 The compound according to any one of claims 1-13, wherein A 4 is CR 8 or N, wherein R 8 is selected from hydrogen, halogen, amino, substituted amino, C 1-3 alkoxy or C 5 -6 heterocyclyl.
  15. 根据权利要求1-14任一项所述的化合物，其特征在于，A 4为CR 8或N，其中R 8选自氢、F、Cl、氨基、-NHCH 3、-N(CH 3) 2、
    Figure PCTCN2020072774-appb-100003
    或甲氧基。     The compound according to any one of claims 1-14, wherein A 4 is CR 8 or N, wherein R 8 is selected from hydrogen, F, Cl, amino, -NHCH 3 , -N(CH 3 ) 2 ,
    Figure PCTCN2020072774-appb-100003
    Or methoxy.
  16. 根据权利要求1-15任一项所述的化合物，其特征在于，U为CH 2或O。 The compound of any one of claims 1-15, wherein U is CH 2 or O.
  17. 根据权利要求1-16任一项所述的化合物，其特征在于，环A任意地选自苯基或C 5-6杂芳基，所述C 5-6杂芳基含有一个或两个N或S杂原子。 The compound according to any one of claims 1-16, wherein ring A is arbitrarily selected from phenyl or C 5-6 heteroaryl, and the C 5-6 heteroaryl contains one or two N Or S heteroatom.
  18. 根据权利要求1-17任一项所述的化合物，其特征在于，Rx任意地选自氢、羟基、卤素、氰基、C 1-3烷基、卤素取代的C 1-3烷基或C 1-3烷氧基。 The compound according to any one of claims 1-17, wherein Rx is arbitrarily selected from hydrogen, hydroxyl, halogen, cyano, C 1-3 alkyl, halogen substituted C 1-3 alkyl or C 1-3 Alkoxy.
  19. 根据权利要求1-18任一项所述的化合物，其特征在于，Rx任意地选自氢、OH、F、Cl、Br、-CN、三氟甲基或甲氧基。The compound according to any one of claims 1-18, wherein Rx is arbitrarily selected from hydrogen, OH, F, Cl, Br, -CN, trifluoromethyl or methoxy.
  20. 根据权利要求1-19任一项所述的化合物，其特征在于，
    Figure PCTCN2020072774-appb-100004
    选自
    Figure PCTCN2020072774-appb-100005
    Figure PCTCN2020072774-appb-100006
    The compound according to any one of claims 1-19, wherein:
    Figure PCTCN2020072774-appb-100004
    Selected from
    Figure PCTCN2020072774-appb-100005
    Figure PCTCN2020072774-appb-100006
  21. 根据权利要求1所述的化合物，或其药学上可接受的盐、互变异构体、溶剂化物、螯合物、非共价复合物或前体药物，其特征在于，所述化合物如式II所示：The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is of the formula II shows:
    Figure PCTCN2020072774-appb-100007
    Figure PCTCN2020072774-appb-100007
    其中，among them,
    R 3任意地选自氢、卤素、氨基、-C(O)NH 2、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基或含取代基的C 1-8烷氧基； Optionally R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
    A 1任意地选自CR 5或N； A 1 is arbitrarily selected from CR 5 or N;
    U任意地选自C(R 9) 2、O或NR 10； U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
    其中，R 5、R 9或R 10独立地选自氢、羟基、卤素、氨基、含取代基的氨基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基或含取代基的C 2-8炔基或C 5-6杂环；或者， Wherein, R 5 , R 9 or R 10 are independently selected from hydrogen, hydroxy, halogen, amino, substituted amino, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy, C substituent group containing 1-8 alkoxy, C 2-8 alkenyl, C 2-8 alkenyl-containing substituent, C 2-8 alkynyl group or a substituted group containing C 2-8 alkynyl Group or C 5-6 heterocycle; or,
    L选自S；L is selected from S;
    R x任意地选自氢、羟基、卤素、氰基、氨基、含取代基的氨基、磺酰基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 3-8环烷基或含取代基的C 3-8环烷基； R x is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy group, a substituted group containing C 1-8 alkoxy, C 3-8 cycloalkyl or C 3-8 cycloalkyl group having substituent;
    n为0、1、2、3或4。n is 0, 1, 2, 3, or 4.
  22. 根据权利要求21所述的化合物，其特征在于，R 3任意地选自氢或甲基。 The compound of claim 21, wherein R 3 is arbitrarily selected from hydrogen or methyl.
  23. 根据权利要求21或22所述的化合物，其特征在于，A 1为CR 5，其中R 5选自F或Cl。 The compound of claim 21 or 22, wherein A 1 is CR 5 , wherein R 5 is selected from F or Cl.
  24. 根据权利要求21或22所述的化合物，其特征在于，A 1为N。 The compound of claim 21 or 22, wherein A 1 is N.
  25. 根据权利要求21-24任一项所述的化合物，其特征在于，U为O或CH 2。 The compound according to any one of claims 21-24, wherein U is O or CH 2 .
  26. 根据权利要求21-25任一项所述的化合物，其特征在于，R x任意地选自氢、羟基、卤素、氰基、C 1-3烷基、卤素取代的C 1-3烷基或C 1-3烷氧基。 The compound according to any one of claims 21-25, wherein R x is optionally selected from hydrogen, hydroxyl, halogen, cyano, C 1-3 alkyl, halogen substituted C 1-3 alkyl or C 1-3 alkoxy.
  27. 根据权利要求21-26任一项所述的化合物，其特征在于，R x任意地选自氢、OH、CN、Cl、F、三氟甲基或甲氧基。 The compound according to any one of claims 21-26, wherein R x is arbitrarily selected from hydrogen, OH, CN, Cl, F, trifluoromethyl or methoxy.
  28. 根据权利要求21-27任一项所述的化合物，其特征在于，n为0、1或2。The compound of any one of claims 21-27, wherein n is 0, 1, or 2.
  29. 根据权利要求1所述的化合物，或其药学上可接受的盐、互变异构体、溶剂化物、螯合物、非共价复合物或前体药物，其特征在于，所述化合物如式III所示：The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, wherein the compound is of the formula As shown in III:
    Figure PCTCN2020072774-appb-100008
    Figure PCTCN2020072774-appb-100008
    R 3任意地选自氢、卤素、氨基、-C(O)NH 2、-C≡N、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基或含取代基的C 1-8烷氧基； Optionally R 3 is selected from hydrogen, halogen, amino, -C (O) NH 2, -C≡N, hydroxy, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy Oxy or C 1-8 alkoxy containing substituents;
    R 4任意地选自氢、卤素、氨基、酰胺基、-C≡N、羧基、羟基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基或含取代基的C 2-8炔基； Optionally R 4 is selected from hydrogen, halo, amino, amido, -C≡N, carboxyl, hydroxyl, C 1-8 alkyl, substituted group containing C 1-8 alkyl, C 1-8 alkoxy, A substituted C 1-8 alkoxy group, a C 2-8 alkenyl group, a substituted C 2-8 alkenyl group, a C 2-8 alkynyl group or a substituted C 2-8 alkynyl group;
    A 1任意地选自CR 5或N； A 1 is arbitrarily selected from CR 5 or N;
    A 2任意地选自CR 6或N； A 2 is arbitrarily selected from CR 6 or N;
    A 3任意地选自CR 7或N； A 3 is arbitrarily selected from CR 7 or N;
    U任意地选自C(R 9) 2、O或NR 10； U is arbitrarily selected from C(R 9 ) 2 , O or NR 10 ;
    其中，R 5、R 6、R 7、R 8、R 9或R 10独立地选自氢、羟基、卤素、氨基、含取代基的氨基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 2-8烯基、含取代基的C 2-8烯基、C 2-8炔基、含取代基的C 2-8炔基或C 5-6杂环基； Wherein, R 5 , R 6 , R 7 , R 8 , R 9 or R 10 are independently selected from hydrogen, hydroxyl, halogen, amino, substituted amino, C 1-8 alkyl, and substituted C 1 -8 alkyl, C 1-8 alkoxy, substituted C 1-8 alkoxy, C 2-8 alkenyl, substituted C 2-8 alkenyl, C 2-8 alkynyl, C 2-8 alkynyl or C 5-6 heterocyclic group containing substituents;
    L选自S；L is selected from S;
    环A任意地选自C 6-10芳基或C 5-10杂芳基，所述C 5-10杂芳基含有一个或两个N或S杂原子； Ring A is optionally selected from a C 6-10 aryl group or a C 5-10 heteroaryl group, the C 5-10 heteroaryl group contains one or two N or S heteroatoms;
    Rx任意地选自氢、羟基、卤素、氰基、氨基、含取代基的氨基、磺酰基、C 1-8烷基、含取代基的C 1-8烷基、C 1-8烷氧基、含取代基的C 1-8烷氧基、C 3-8环烷基或含取代基的C 3-8环烷基； Rx is optionally selected from hydrogen, hydroxyl, halogen, cyano, amino, substituted amino, sulfonyl, C 1-8 alkyl, substituted C 1-8 alkyl, C 1-8 alkoxy containing substituted C 1-8 alkoxy group, C 3-8 cycloalkyl or an unsubstituted C 3-8 cycloalkyl group;
    n为0、1、2、3或4。n is 0, 1, 2, 3, or 4.
  30. 根据权利要求29所述的化合物，其特征在于，R 3选自氢。 The compound of claim 29, wherein R 3 is selected from hydrogen.
  31. 根据权利要求29或30所述的化合物，其特征在于，R 4任意地选自氢或Cl。 The compound according to claim 29 or 30, wherein R 4 is arbitrarily selected from hydrogen or Cl.
  32. 根据权利要求29-31任一项所述的化合物，其特征在于，A 1为CR 5或N，其中R 5选自氢、Cl或三氟甲基。 The compound according to any one of claims 29-31, wherein A 1 is CR 5 or N, wherein R 5 is selected from hydrogen, Cl or trifluoromethyl.
  33. 根据权利要求29-32任一项所述的化合物，其特征在于，A 2为CR 6或N，其中R 6选自Cl、氨基、
    Figure PCTCN2020072774-appb-100009
    或-O-CH 3。     The compound according to any one of claims 29-32, wherein A 2 is CR 6 or N, wherein R 6 is selected from Cl, amino,
    Figure PCTCN2020072774-appb-100009
    Or -O-CH 3 .
  34. 根据权利要求29-33任一项所述的化合物，其特征在于，A 3为CR 7或N，其中R 7为氢。 The compound of any one of claims 29-33, wherein A 3 is CR 7 or N, wherein R 7 is hydrogen.
  35. 根据权利要求29-34任一项所述的化合物，其特征在于，U为CH 2。 The compound of any one of claims 29-34, wherein U is CH 2 .
  36. 根据权利要求29-35任一项所述的化合物，其特征在于，环A任意地选自苯基或C 5-6杂芳基，所述C 5-6杂芳基含有一个或两个N或S杂原子。 The compound according to any one of claims 29-35, wherein ring A is optionally selected from phenyl or C 5-6 heteroaryl, said C 5-6 heteroaryl contains one or two N Or S heteroatom.
  37. 根据权利要求29-36任一项所述的化合物，其特征在于，Rx任意地选自氢、Cl、Br、三氟甲基或甲氧基。The compound according to any one of claims 29-36, wherein Rx is arbitrarily selected from hydrogen, Cl, Br, trifluoromethyl or methoxy.
  38. 根据权利要求29-37任一项所述的化合物，其特征在于，
    Figure PCTCN2020072774-appb-100010
    选自
    Figure PCTCN2020072774-appb-100011
    Figure PCTCN2020072774-appb-100012
    The compound according to any one of claims 29-37, wherein:
    Figure PCTCN2020072774-appb-100010
    Selected from
    Figure PCTCN2020072774-appb-100011
    Figure PCTCN2020072774-appb-100012
  39. 根据权利要求1所述的化合物，其特征在于，所述化合物是：The compound of claim 1, wherein the compound is:
    2)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫代)-5-甲基吡嗪-2-基)甲醇；2) (S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)-5-methylpyrazin-2-yl)methanol;
    3)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲腈；3) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine) -4-yl)thio)pyrazine-2-carbonitrile;
    4)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫基)吡嗪-2-甲酰胺；4)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorophenyl )Thio)pyrazine-2-carboxamide;
    5)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲酰胺；5) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine) -4-yl)thio)pyrazine-2-carboxamide;
    7)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-甲氧基吡啶-4-基)硫基)吡嗪-2-甲酰胺；7) (S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methoxy (Pyridin-4-yl)thio)pyrazine-2-carboxamide;
    8)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-甲酰胺；8)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl) Pyridin-3-yl)thio)pyrazine-2-carboxamide;
    9)(S)-3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-甲酰胺；9)(S)-3-(1-Amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2- (Trifluoromethyl)pyridin-3-yl)thio)pyrazine-2-carboxamide;
    10)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-溴-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；10) (S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-bromo-1,3-dihydrospiro(indene-2, 4'-Piperidine]-1'-yl)pyrazine-2-carboxamide;
    11)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；11) (S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-methoxy-1,3-dihydrospiro(indene- 2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
    12)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；12) (S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-methoxy-1,3-dihydrospiro(indene- 2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
    13)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-5,6-二甲基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；13)(S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro[ Indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
    14)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；14)(S)-6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-(trifluoromethyl)-1,3-dihydrospiro [Indene-2,4'-piperidine]-1'-yl)pyrazine-2-carboxamide;
    15)(S)-6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氯-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-甲酰胺；15) (S)-6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro[indene-2, 4'-Piperidine]-1'-yl)pyrazine-2-carboxamide;
    16)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((6-氨基-3-氯吡啶-2-基)硫基)吡嗪-2-甲酰胺；16)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((6-amino-3-chloropyridine -2-yl)thio)pyrazine-2-carboxamide;
    17)(S)-3-(5-氨基-2-甲氧基-5,7-二氢螺[环戊二烯[b]吡啶-6,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲酰胺；17)(S)-3-(5-Amino-2-methoxy-5,7-dihydrospiro[cyclopentadiene[b]pyridine-6,4'-piperidine]-1'-yl) -6-((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide;
    18)(S)-3-(4-氨基-2-氯-4,6-二氢螺[环戊二烯并[d]噻唑-5,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-甲酰胺；18)(S)-3-(4-Amino-2-chloro-4,6-dihydrospiro[cyclopenta[d]thiazole-5,4'-piperidine]-1'-yl)- 6-((2-Amino-3-chloropyridin-4-yl)thio)pyrazine-2-carboxamide;
    19)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氟-3-甲氧基苯基)硫基)吡嗪-2-基)甲醇；19)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-fluoro-3-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;
    20)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-甲氧基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；20)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-methoxypyrimidine- 4-yl)thio)pyrazin-2-yl)methanol;
    21)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)甲醇；21)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloro Pyridin-4-yl)thio)pyrazin-2-yl)methanol;
    22)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；22)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-4- (Yl)thio)pyrazin-2-yl)methanol;
    23)(S)-1-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)乙-1-酮；23)(S)-1-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)ethan-1-one;
    24)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；24)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-( (1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol;
    25)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-(三氟甲基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；25)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-(trifluoromethyl )Pyridin-4-yl)thio)pyrazin-2-yl)methanol;
    26)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；26)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-(trifluoromethyl)-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
    27)1-(3-((S)-1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)乙-1-醇；27) 1-(3-((S)-1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)ethan-1-ol;
    28)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((5-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；28)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((5-chloro-2-( (1-methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol;
    29)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(二甲基氨基)-3-氟吡啶-4-基)硫基)吡嗪-2-基)甲醇；29)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(dimethylamino )-3-fluoropyridin-4-yl)thio)pyrazin-2-yl)methanol;
    30)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-吡啶]-1'-基)-6-((3-氟-2-(甲基氨基)吡啶-4-基)硫基)吡嗪-2-基)甲醇；30)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-pyridine]-1'-yl)-6-((3-fluoro-2-(methyl (Amino)pyridin-4-yl)thio)pyrazin-2-yl)methanol;
    31)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(二氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲基；31)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(difluoromethyl )Pyridin-3-yl)thio)pyrazin-2-yl)methyl;
    32)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-(二甲基氨基)吡啶-4-基)硫基)吡嗪-2-基)甲基；32)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-( Dimethylamino)pyridin-4-yl)thio)pyrazin-2-yl)methyl;
    33)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氟-2-甲氧基苯基)硫基)吡嗪-2-基)甲醇；33)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-fluoro-2-methyl (Oxyphenyl)thio)pyrazin-2-yl)methanol;
    34)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-5-氟-2-甲氧基苯基)硫基)吡嗪-2-基)甲醇；34)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-5-fluoro -2-Methoxyphenyl)thio)pyrazin-2-yl)methanol;
    35)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(喹啉-4-基硫基)吡嗪-2-基)甲醇；35)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(quinolin-4-ylsulfanyl )Pyrazin-2-yl)methanol;
    36)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲醇；36)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl )Pyridin-3-yl)thio)pyrazin-2-yl)methanol;
    37)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氯苯基)硫基)吡嗪-2-基)甲醇；37)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dichlorobenzene (Yl)thio)pyrazin-2-yl)methanol;
    38)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(三氟甲基)吡啶-3-基)硫基)吡嗪-2-基)甲醇；38)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(trifluoromethyl )Pyridin-3-yl)thio)pyrazin-2-yl)methanol;
    39)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氟吡啶-4-基)硫基)吡嗪-2-基)甲基；39)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-fluoro Pyridin-4-yl)thio)pyrazin-2-yl)methyl;
    40)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-甲氧基吡啶-4-基)硫基)吡嗪-2-基)甲醇；40)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-methyl Oxypyridin-4-yl)thio)pyrazin-2-yl)methanol;
    41)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氟-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；41)(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-fluoro-1,3-dihydrospiro(indene-2 ,4'-Piperidine]-1'-yl)pyrazin-2-yl)methanol;
    42)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；42)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-6-methoxy-1,3-dihydrospiro(indene -2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
    43)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-5,6-二甲基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；43)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-5,6-dimethyl-1,3-dihydrospiro [Indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
    44)(R)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(3-氨基-3H-螺[苯并呋喃-2,4'-哌啶]-1'-基)吡嗪-2-基)甲基；44)(R)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(3-amino-3H-spiro[benzofuran-2,4'-piperidine ]-1'-yl)pyrazin-2-yl)methyl;
    45)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-4-腈；45)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-4-carbonitrile;
    46)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-6-氯-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；46)(S)-(6-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(1-amino-6-chloro-1,3-dihydrospiro(indene-2 ,4'-Piperidine]-1'-yl)pyrazin-2-yl)methanol;
    47)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(吡啶-4-基硫基)吡嗪-2-基)甲醇；47)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(pyridin-4-ylsulfanyl) Pyrazin-2-yl)methanol;
    48)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(5-氨基-5,7-二氢螺[环戊烯并[b]吡啶-6,4'-哌啶]-1'-基)吡嗪-2-基)甲基；48)(S)-(6-((2-amino-3-chloropyridin-4-yl)thio)-3-(5-amino-5,7-dihydrospiro[cyclopenteno[b] Pyridine-6,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl;
    49)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-4-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲醇；49)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-4-(trifluoromethyl)-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methanol;
    50)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-6-醇；50)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-6-ol;
    51)(S)-4-((5-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(羟基甲基)吡嗪-2-基)硫基)嘧啶-2-醇；51)(S)-4-((5-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(hydroxymethyl)pyridine (Azin-2-yl)thio)pyrimidin-2-ol;
    52)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-7-氟-1,3-二氢螺[茚-2,4'-哌啶]-4-醇；52)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 7-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-4-ol;
    53)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2,3-二氢-[1,4]二氧杂环己并[2,3-b]吡啶-8-基)硫基)吡嗪-2-基)甲醇；53)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2,3-dihydro- [1,4]Dioxo[2,3-b]pyridin-8-yl)thio)pyrazin-2-yl)methanol;
    54)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-6-腈；54)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-6-nitrile;
    55)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-4-氟-1,3-二氢螺[茚-2,4'-哌啶]-7-腈；55)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 4-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-7-nitrile;
    56)(S)-(6-((2-氨基-3-氯吡啶-4-基)硫基)-3-(1-氨基-7-(三氟甲基)-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)吡嗪-2-基)甲基；56)(S)-(6-((2-Amino-3-chloropyridin-4-yl)thio)-3-(1-amino-7-(trifluoromethyl)-1,3-dihydro Spiro[indene-2,4'-piperidine]-1'-yl)pyrazin-2-yl)methyl;
    57)(S)-1-氨基-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(羟基甲基)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-7-醇；57)(S)-1-amino-1'-(5-((2-amino-3-chloropyridin-4-yl)sulfanyl)-3-(hydroxymethyl)pyrazin-2-yl)- 1,3-Dihydrospiro[indene-2,4'-piperidine]-7-ol;
    58)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氟吡啶-4-基)硫基)吡嗪-2-甲酰胺；58)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-fluoropyridine -4-yl)thio)pyrazine-2-carboxamide;
    59)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((3-氯-2-((1-甲基-1H-吡唑-5-基)氨基)吡啶-4-基)硫基)吡嗪-2-甲酰胺；59)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((3-chloro-2-(( 1-Methyl-1H-pyrazol-5-yl)amino)pyridin-4-yl)thio)pyrazine-2-carboxamide;
    60)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)-N-羟基吡嗪-2-甲酰胺；60)(S)-3-(1-Amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3-chloropyridine -4-yl)thio)-N-hydroxypyrazine-2-carboxamide;
    61)1-(3-((S)-1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)-2,2,2-三氟乙-1-醇；61)1-(3-((S)-1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)-2,2,2-trifluoroethane-1-ol;
    62)(S)-1'-(5-((2-氨基-3-氯吡啶-4-基)硫基)-3-(甲氧基甲基l)吡嗪-2-基)-1,3-二氢螺[茚-2,4'-哌啶]-1-胺；62)(S)-1'-(5-((2-Amino-3-chloropyridin-4-yl)sulfanyl)-3-(methoxymethyll)pyrazin-2-yl)-1 ,3-Dihydrospiro[indene-2,4'-piperidine]-1-amine;
    63)(S)-2-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基-3-氯吡啶-4-基)硫基)吡嗪-2-基)丙-2-醇；63)(S)-2-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino-3 -Chloropyridin-4-yl)thio)pyrazin-2-yl)propan-2-ol;
    64)(S)-(3-(1-氨基-6-甲氧基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；64)(S)-(3-(1-amino-6-methoxy-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2 -Aminopyrimidin-4-yl)thio)pyrazin-2-yl)methanol;
    65)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-(嘧啶-4-基硫基)吡嗪-2-基)甲醇；65)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-(pyrimidin-4-ylthio) Pyrazin-2-yl)methanol;
    66)(S)-(3-(1-氨基-5-氯-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；66)(S)-(3-(1-Amino-5-chloro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;
    67)(S)-(3-(1-氨基-5-氟-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-基)甲醇；67)(S)-(3-(1-amino-5-fluoro-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-amino Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;
    68)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-(甲基氨基)嘧啶-4-基)硫基)吡嗪-2-基)甲醇；68)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-(methylamino) Pyrimidine-4-yl)thio)pyrazin-2-yl)methanol;
    69)(S)-(3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)-5-甲基哌嗪-2-基)甲醇；或69)(S)-(3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidine]-1'-yl)-6-((2-aminopyrimidine-4- Yl)thio)-5-methylpiperazin-2-yl)methanol; or
    70)(S)-3-(1-氨基-1,3-二氢螺[茚-2,4'-哌啶]-1'-基)-6-((2-氨基嘧啶-4-基)硫基)吡嗪-2-甲酰胺。70)(S)-3-(1-amino-1,3-dihydrospiro[indene-2,4'-piperidin]-1'-yl)-6-((2-aminopyrimidin-4-yl ) Thio) pyrazine-2-carboxamide.
  40. 一种药物组合物，其特征在于，包含治疗有效量的至少一种权利要求1-38任一项所述的化合物和至少一种药学上可接受的辅料。A pharmaceutical composition characterized by comprising a therapeutically effective amount of at least one compound according to any one of claims 1-38 and at least one pharmaceutically acceptable excipient.
  41. 根据权利要求40所述的药物组合物，其特征在于，所述的化合物和药学上可接受的辅料的质量百分比为0.0001:1-10。The pharmaceutical composition according to claim 40, wherein the mass percentage of the compound and pharmaceutically acceptable excipients is 0.0001:1-10.
  42. 权利要求1-39任一项所述的化合物或权利要求40或41所述的药物组合物在制备药物中的应用。Use of the compound of any one of claims 1-39 or the pharmaceutical composition of claim 40 or 41 in the preparation of medicines.
  43. 根据权利要求42所述的应用，其特征在于，所述药物用于治疗、预防、延迟或阻止癌症，癌症转移，心血管疾病，免疫疾病，纤维化或眼部疾病。The application according to claim 42, wherein the medicine is used to treat, prevent, delay or stop cancer, cancer metastasis, cardiovascular disease, immune disease, fibrosis or eye disease.
  44. 权利要求1-39任一项所述的化合物或权利要求40或41所述的药物组合物在制备治疗由SHP2介导的疾病的药物中的应用。Use of the compound of any one of claims 1-39 or the pharmaceutical composition of claim 40 or 41 in the preparation of a medicine for treating diseases mediated by SHP2.
  45. 根据权利要求44所述的应用，其特征在于，所述疾病是癌症。The use according to claim 44, wherein the disease is cancer.
  46. 根据权利要求43或45所述的应用，其特征在于，所述癌症选自Noonan综合征、豹斑综合征、青少年髓单核细胞白血病、神经母细胞瘤、黑色素瘤、头颈部鳞状细胞癌、急性髓性白血病、乳腺癌、食道肿瘤、肺癌、结肠癌、头癌、胃癌、淋巴瘤、胶质母细胞瘤、胰腺癌或其组合。The use according to claim 43 or 45, wherein the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell Cancer, acute myeloid leukemia, breast cancer, esophageal tumor, lung cancer, colon cancer, head cancer, gastric cancer, lymphoma, glioblastoma, pancreatic cancer, or a combination thereof.
  47. 根据权利要求42所述的应用，其特征在于，所述的药物用作SHP2抑制剂。The use according to claim 42, wherein the drug is used as an SHP2 inhibitor.
  48. 一种治疗和/或预防由SHP2介导的疾病的方法，其特征在于，向治疗对象施用治疗有效量的权利要求1-39任一项所述的化合物或权利要求40或41所述的药物组合物。A method for treating and/or preventing diseases mediated by SHP2, characterized in that a therapeutically effective amount of the compound according to any one of claims 1-39 or the drug according to claim 40 or 41 is administered to a subject to be treated combination.
  49. 根据权利要求48所述的方法，其特征在于，所述SHP2介导的疾病是癌症。The method of claim 48, wherein the SHP2-mediated disease is cancer.
  50. 根据权利要求49所述的方法，其特征在于，所述癌症选自Noonan综合征、豹斑综合征、青少年髓单核细胞白血病、神经母细胞瘤、黑色素瘤、头颈部鳞状细胞癌、急性髓性白血病、乳腺癌、食道肿瘤、肺癌、结肠癌、头癌、胃癌、淋巴瘤、胶质母细胞瘤、胰腺癌或其组合。The method of claim 49, wherein the cancer is selected from Noonan syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, Acute myeloid leukemia, breast cancer, esophageal tumor, lung cancer, colon cancer, head cancer, gastric cancer, lymphoma, glioblastoma, pancreatic cancer, or a combination thereof.
  51. 一种治疗癌症的方法，包括向治疗对象施用治疗有效量的权利要求1-38任一项所述的化合物或权利要求40或41所述的药物组合物，其特征在于，所述癌症是Noonan综合征、豹斑综合征、青少年髓单核细胞白血病、神经母细胞瘤、黑色素瘤、头颈部鳞状细胞癌、急性髓性白血病、乳腺癌、食道肿瘤、肺癌、结肠癌、头癌、胃癌、淋巴瘤、胶质母细胞瘤、胰腺癌或其组合。A method for treating cancer, comprising administering a therapeutically effective amount of the compound of any one of claims 1-38 or the pharmaceutical composition of claim 40 or 41 to a subject, wherein the cancer is Noonan Syndrome, leopard spot syndrome, juvenile myelomonocytic leukemia, neuroblastoma, melanoma, head and neck squamous cell carcinoma, acute myeloid leukemia, breast cancer, esophageal tumor, lung cancer, colon cancer, head cancer, Gastric cancer, lymphoma, glioblastoma, pancreatic cancer, or a combination thereof.
  52. 根据权利要求48-51任一项所述的方法，其特征在于，所述治疗对象为人类。The method according to any one of claims 48-51, wherein the subject to be treated is a human.
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