WO2023274280A1 - Crystal form of biphenyl derivative inhibitor and preparation method therefor - Google Patents

Crystal form of biphenyl derivative inhibitor and preparation method therefor Download PDF

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WO2023274280A1
WO2023274280A1 PCT/CN2022/102195 CN2022102195W WO2023274280A1 WO 2023274280 A1 WO2023274280 A1 WO 2023274280A1 CN 2022102195 W CN2022102195 W CN 2022102195W WO 2023274280 A1 WO2023274280 A1 WO 2023274280A1
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methyl
biphenyl
imidazo
dichloro
tetrahydro
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PCT/CN2022/102195
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French (fr)
Chinese (zh)
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詹小兰
呙临松
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上海翰森生物医药科技有限公司
江苏豪森药业集团有限公司
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Priority to CN202280045197.3A priority Critical patent/CN117561256A/en
Publication of WO2023274280A1 publication Critical patent/WO2023274280A1/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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • 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
    • C07D471/04—Ortho-condensed systems

Definitions

  • the invention belongs to the technical field of medicinal chemistry, and in particular relates to a crystal form of a biphenyl derivative inhibitor, a preparation method and application thereof.
  • the immune system plays an important role in controlling many diseases, including cancer.
  • tumor cells evade immune attack or suppress the activation of the immune system in various ways.
  • PD-1 programmed cell death protein 1
  • PD-1 a member of the CD28 superfamily, is an immunosuppressive receptor on the surface of immune cells, especially cytotoxic T cells.
  • PD-1 has two ligands PD-L1 and PD-L2, among which PD-L1, programmed cell death receptor-ligand 1, is expressed in a variety of cells, such as macrophages and dendritic cells , and generally highly expressed on tumor cells.
  • PD-L1 plays an immunosuppressive role by combining with PD-1 and makes tumor cells evade the killing of T cells, inhibits the activation of T cells and the production of corresponding cytokines, weakens infectious immunity and tumor immunity, and promotes the development of infectious diseases and tumors. progress.
  • the use of PD-L1 inhibitors such as antibodies or small molecule inhibitors can relieve the immunosuppressive effect of PD-L1 and promote the immune clearance of tumors, thereby achieving the effect of treating tumors.
  • PD-1/PD-L1 is a hot spot in the research of tumor immunotherapy in recent years.
  • the breadth, depth and durability of the monoclonal antibody drug response are very rare.
  • PD-1/PD-L1 monoclonal antibody drugs in clinical practice. marketed and achieved great success.
  • PD-L1 inhibitors can be used to treat almost all major cancers, including non-small cell lung cancer, liver cancer, gastric cancer, intestinal cancer, kidney cancer, etc., and have great clinical application value.
  • PD-L1 inhibitors from macromolecules to small molecules are becoming a new research and development trend and hotspot.
  • Small molecule inhibitors have many natural advantages in terms of administration methods and production costs, and have the potential to replace antibody macromolecules, including BMS at present.
  • Foreign pharmaceutical companies including Incyte and other companies are actively developing PD-L1 small molecule inhibitors.
  • the oral small-molecule inhibitor developed by BMS is currently in the preclinical research stage, and several patents have been published consecutively.
  • the small-molecule inhibitor INCB086550 developed by Incyte is in the first phase of clinical research.
  • the small-molecule inhibitor developed by Aurigene/Curis CA-170 is in Phase II clinical research.
  • Small molecule inhibitors of PD-L1 have good application prospects as drugs in the pharmaceutical industry.
  • small molecule inhibitors of PD-L1 can be administered orally, which has the advantage of stronger compliance than intravenous administration of antibody drugs, and can avoid antibody Serious side effects such as colitis caused by long-term residence in the body.
  • small molecule inhibitors of PD-L1 have a unique mechanism of action that binds to PD-L1 and endocytizes it, which may show different efficacy from antibodies in clinical practice.
  • the production and quality control costs of PD-L1 small molecule inhibitors are lower, and they have a price advantage that is far lower than the cost of macromolecular drugs.
  • PD-L1 antibody inhibitors they can be applied to a variety of major tumors and have a huge market potential.
  • the object of the present invention is to provide a kind of crystal form of the compound shown in general formula (I), its structure is as follows:
  • Ring A is selected from 4-8 membered heterocyclic groups; preferably 4-membered, 7-membered or 8-membered heterocyclic groups;
  • R 1 is selected from hydrogen, deuterium, halogen, hydroxyl, cyano, carboxyl, aldehyde, oxo, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O)R a , -C( O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a , wherein the C 1 -6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl and C 3-8 cycloalkyl,
  • R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, carboxyl, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 Haloalkyl or C 3-8 cycloalkyl, wherein said C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl and C 3-8 cycloalkyl, optionally further One or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-6 alkyl;
  • n 0, 1 or 2;
  • x 0, 1 or 2.
  • M is O, -NR 2 or -CR 3 R 4 ;
  • R 2 is selected from hydrogen, deuterium, aldehyde, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 3-6 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O)R a , -C(O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a ;
  • R 3 and R 4 are each independently selected from hydrogen, deuterium, halogen, hydroxyl, cyano, carboxyl, aldehyde, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 Haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 3-6 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O) R a , -C(O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a , The C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl and C 3-6 cycloalkyl
  • R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, carboxyl, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 Haloalkyl or C 3-6 cycloalkyl, wherein said C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl and C 3-6 cycloalkyl, optionally further One or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-3 alkyl;
  • n 0, 1 or 2.
  • the crystal form of the compound is the crystal form of the following compound:
  • the crystal form is the compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Crystalline form of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide.
  • the X-ray powder diffraction pattern of the crystal form A has a diffraction peak at 2 ⁇ of 7.8 ⁇ 0.2°; or has a diffraction peak at 14.1 ⁇ 0.2°; or has a diffraction peak at 15.3 ⁇ 0.2°; or has a diffraction peak at 19.5 ⁇ 0.2°; Diffraction peak at 0.2°; or diffraction peak at 12.0 ⁇ 0.2°; or diffraction peak at 12.4 ⁇ 0.2°; or diffraction peak at 13.2 ⁇ 0.2°; or diffraction peak at 14.6 ⁇ 0.2° or have a diffraction peak at 21.2 ⁇ 0.2°; or have a diffraction peak at 22.0 ⁇ 0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
  • the X-ray powder diffraction pattern of the crystal form B has a diffraction peak at 2 ⁇ of 4.2 ⁇ 0.2°; or has a diffraction peak at 12.2 ⁇ 0.2°; or has a diffraction peak at 12.6 ⁇ 0.2°; or has a diffraction peak at 16.5 ⁇ 0.2°; Diffraction peak at 0.2°; or diffraction peak at 16.8 ⁇ 0.2°; or diffraction peak at 18.4 ⁇ 0.2°; or diffraction peak at 18.9 ⁇ 0.2°; or diffraction peak at 21.1 ⁇ 0.2° or have a diffraction peak at 22.2 ⁇ 0.2°; or have a diffraction peak at 22.4 ⁇ 0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
  • the X-ray powder diffraction pattern of the crystal form C has a diffraction peak at 2 ⁇ of 11.4 ⁇ 0.2°; or has a diffraction peak at 12.5 ⁇ 0.2°; or has a diffraction peak at 21.1 ⁇ 0.2°; or has a diffraction peak at 23.0 ⁇ 0.2°; Diffraction peak at 0.2°; or diffraction peak at 26.1 ⁇ 0.2°; or diffraction peak at 26.6 ⁇ 0.2°; or diffraction peak at 13.0 ⁇ 0.2°; or diffraction peak at 14.0 ⁇ 0.2° or have a diffraction peak at 14.7 ⁇ 0.2°; or have a diffraction peak at 15.8 ⁇ 0.2°; preferably include any 2-5 of the above-mentioned diffraction peaks, or 3-5, or 3-6, or 3 -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
  • the X-ray powder diffraction pattern of the crystal form D has a diffraction peak at 2 ⁇ of 9.5 ⁇ 0.2°; or has a diffraction peak at 10.4 ⁇ 0.2°; or has a diffraction peak at 14.5 ⁇ 0.2°; or has a diffraction peak at 19.9 ⁇ 0.2°; Diffraction peak at 0.2°; or diffraction peak at 20.8 ⁇ 0.2°; or diffraction peak at 24.8 ⁇ 0.2°; or diffraction peak at 11.7 ⁇ 0.2°; or diffraction peak at 13.3 ⁇ 0.2° or have a diffraction peak at 17.2 ⁇ 0.2°; or have a diffraction peak at 23.8 ⁇ 0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places.
  • the X-ray powder diffraction pattern of the crystal form A contains at least one or more diffraction peaks at 2 ⁇ of 7.8 ⁇ 0.2°, 14.1 ⁇ 0.2°, and 15.3 ⁇ 0.2°, Preferably two of them are included, more preferably three; optionally, at least one of 19.5 ⁇ 0.2°, 12.0 ⁇ 0.2°, 12.4 ⁇ 0.2°, 13.2 ⁇ 0.2°, 14.6 ⁇ 0.2° can be included, preferably 2, 3, 4 or 5 of them;
  • the X-ray powder diffraction pattern of the crystal form B includes at least one or more diffraction peaks located at 2 ⁇ of 4.2 ⁇ 0.2°, 12.2 ⁇ 0.2°, and 12.6 ⁇ 0.2°, preferably two of them, and more It preferably contains three; optionally, it can further include at least one of 16.5 ⁇ 0.2°, 16.8 ⁇ 0.2°, 18.4 ⁇ 0.2°, 18.9 ⁇ 0.2°, 21.1 ⁇ 0.2°, preferably 2, 3, 4 or 5;
  • the X-ray powder diffraction pattern of the crystal form C contains at least one or more diffraction peaks located at 2 ⁇ of 11.4 ⁇ 0.2°, 12.5 ⁇ 0.2°, and 21.1 ⁇ 0.2°, preferably two of them, and more It preferably contains three; optionally, it can further contain at least one of 23.0 ⁇ 0.2°, 26.1 ⁇ 0.2°, 26.6 ⁇ 0.2°, 13.0 ⁇ 0.2°, 14.0 ⁇ 0.2°, preferably 2, 3, 4 or 5;
  • the X-ray powder diffraction pattern of the crystal form D contains at least one or more diffraction peaks located at 2 ⁇ of 9.5 ⁇ 0.2°, 10.4 ⁇ 0.2°, and 14.5 ⁇ 0.2°, preferably two of them, and more Preferably include three; optionally, it may further include at least one of 19.9 ⁇ 0.2°, 20.8 ⁇ 0.2°, 24.8 ⁇ 0.2°, 11.7 ⁇ 0.2°, 13.3 ⁇ 0.2°, preferably including 2, 3, 4 or 5 bars.
  • the X-ray powder diffraction pattern of the crystal form A optionally further comprises One or more diffraction peaks at 0.2°, 16.7 ⁇ 0.2° or 24.5 ⁇ 0.2°; preferably at least any 2-3, or 4-5, or 6-7 of them; more preferably, including Any 2, 3, 4, 5, 6, 7 places;
  • the X-ray powder diffraction pattern of the crystal form B optionally further includes a position at 2 ⁇ of 22.2 ⁇ 0.2°, 22.4 ⁇ 0.2°, 22.9 ⁇ 0.2°, 23.6 ⁇ 0.2°, 26.6 ⁇ 0.2°, 13.8 ⁇ 0.2° Or one or more diffraction peaks at 14.5 ⁇ 0.2°; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5, 6, 7;
  • the X-ray powder diffraction pattern of the crystalline form C optionally further includes a position at 2 ⁇ of 14.7 ⁇ 0.2°, 15.8 ⁇ 0.2°, 18.0 ⁇ 0.2°, 19.8 ⁇ 0.2°, 23.6 ⁇ 0.2°, 9.1 ⁇ 0.2° Or one or more diffraction peaks at 16.7 ⁇ 0.2°; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5, 6, 7;
  • the X-ray powder diffraction pattern of the crystal form D optionally further includes a position at 2 ⁇ of 17.2 ⁇ 0.2°, 23.8 ⁇ 0.2°, 25.5 ⁇ 0.2°, 27.1 ⁇ 0.2°, 15.3 ⁇ 0.2°, 15.8 ⁇ 0.2° Or one or more diffraction peaks at 21.4 ⁇ 0.2°; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5, 6, 7.
  • the X-ray powder diffraction pattern of the crystalline form A optionally includes 2 ⁇ at 7.8 ⁇ 0.2°, 14.1 ⁇ 0.2°, 15.3 ⁇ 0.2°, 19.5 ⁇ 0.2°, 12.0 ⁇ 0.2° , 12.4 ⁇ 0.2°, 13.2 ⁇ 0.2°, 14.6 ⁇ 0.2°, 21.2 ⁇ 0.2°, 22.0 ⁇ 0.2°, 22.6 ⁇ 0.2°, 25.0 ⁇ 0.2°, 28.3 ⁇ 0.2°, 16.7 ⁇ 0.2° or 24.5 ⁇ 0.2°
  • the X-ray powder diffraction pattern of the crystal form B optionally includes a position at 2 ⁇ of 4.2 ⁇ 0.2°, 12.2 ⁇ 0.2°, 12.6 ⁇ 0.2°, 16.5 ⁇ 0.2°, 16.8 ⁇ 0.2°, 18.4 ⁇ 0.2°, One of 18.9 ⁇ 0.2°, 21.1 ⁇ 0.2°, 22.2 ⁇ 0.2°, 22.4 ⁇ 0.2°, 22.9 ⁇ 0.2°, 23.6 ⁇ 0.2°, 26.6 ⁇ 0.2°, 13.8 ⁇ 0.2° or 14.5 ⁇ 0.2°, or multiple diffraction peaks,
  • the X-ray powder diffraction pattern of the crystal form B has diffraction peaks at the following positions at 2 ⁇ :
  • the X-ray powder diffraction pattern of the crystal form C optionally includes a position at 2 ⁇ of 11.4 ⁇ 0.2°, 12.5 ⁇ 0.2°, 21.1 ⁇ 0.2°, 23.0 ⁇ 0.2°, 26.1 ⁇ 0.2°, 26.6 ⁇ 0.2°, One of 13.0 ⁇ 0.2°, 14.0 ⁇ 0.2°, 14.7 ⁇ 0.2°, 15.8 ⁇ 0.2°, 18.0 ⁇ 0.2°, 19.8 ⁇ 0.2°, 23.6 ⁇ 0.2°, 9.1 ⁇ 0.2° or 16.7 ⁇ 0.2° or multiple diffraction peaks,
  • the X-ray powder diffraction pattern of the crystal form D optionally includes a position at 2 ⁇ of 9.5 ⁇ 0.2°, 10.4 ⁇ 0.2°, 14.5 ⁇ 0.2°, 19.9 ⁇ 0.2°, 20.8 ⁇ 0.2°, 24.8 ⁇ 0.2°, One of 11.7 ⁇ 0.2°, 13.3 ⁇ 0.2°, 17.2 ⁇ 0.2°, 23.8 ⁇ 0.2°, 25.5 ⁇ 0.2°, 27.1 ⁇ 0.2°, 15.3 ⁇ 0.2°, 15.8 ⁇ 0.2° or 21.4 ⁇ 0.2° or multiple diffraction peaks,
  • diffraction peaks at optional 4, 5, 6, 8 or 10 positions.
  • the crystal form is N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methanol Base)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5 , the crystal form B of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
  • the X-ray powder diffraction pattern of the crystal form B has a diffraction peak at 4.0 ⁇ 0.2°; or has a diffraction peak at 12.0 ⁇ 0.2°; or has a diffraction peak at 12.4 ⁇ 0.2°; or has a diffraction peak at 16.4 ⁇ 0.2° or have a diffraction peak at 16.6 ⁇ 0.2°; or have a diffraction peak at 18.2 ⁇ 0.2°; or have a diffraction peak at 18.7 ⁇ 0.2°; or have a diffraction peak at 20.9 ⁇ 0.2°; or Have a diffraction peak at 22.0 ⁇ 0.2°; or have a diffraction peak at 22.3 ⁇ 0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3-8 of the above-mentioned diffraction peaks , or 5-8, or 6-8, more preferably including any 6, 7 or 8 of them;
  • the X-ray powder diffraction pattern of the crystal form B includes at least one or more diffraction peaks at 2 ⁇ of 4.0 ⁇ 0.2°, 12.0 ⁇ 0.2°, and 12.4 ⁇ 0.2°, preferably two of them, More preferably, it contains three; optionally, it can further include at least one of 16.4 ⁇ 0.2°, 16.6 ⁇ 0.2°, 18.2 ⁇ 0.2°, 18.7 ⁇ 0.2°, 20.9 ⁇ 0.2°, preferably 2 or 3 of them , 4 or 5;
  • the X-ray powder diffraction pattern of the crystal form B optionally further comprises One or more diffraction peaks at 0.2° or 14.3 ⁇ 0.2°; preferably at least include any 2-3, or 4-5, or 6-7 of them; more preferably, include any 2 or 3 of them place, 4 places, 5 places, 6 places, 7 places;
  • the X-ray powder diffraction pattern of the crystal form B optionally includes °, 18.7 ⁇ 0.2°, 20.9 ⁇ 0.2°, 22.0 ⁇ 0.2°, 22.3 ⁇ 0.2°, 22.7 ⁇ 0.2°, 23.5 ⁇ 0.2°, 26.5 ⁇ 0.2°, 13.6 ⁇ 0.2° or 14.3 ⁇ 0.2°
  • One or more diffraction peaks preferably, including optional 4, 5, 6, 8 or 10 diffraction peaks;
  • the X-ray powder diffraction pattern of the crystal form B is shown in FIG. 10 .
  • Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form A of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2 ⁇ angles and interplanar spacing d values using Cu-K ⁇ radiation The diffraction peaks are shown in Table 1.
  • Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form B of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2 ⁇ angles and interplanar spacing d values using Cu-K ⁇ radiation The diffraction peaks are shown in Table 2.
  • Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form C of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2 ⁇ angles and interplanar spacing d values using Cu-K ⁇ radiation The diffraction peaks are shown in Table 3.
  • Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form D of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2 ⁇ angles and interplanar spacing d values using Cu-K ⁇ radiation The diffraction peaks are shown in Table 4.
  • Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl) -6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6 , Relative peak intensities in the X-ray powder diffraction patterns of 7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide in Form A, Form B, Form C and Form D
  • the 2 ⁇ error between the positions of the top ten diffraction peaks and the corresponding positions in Figure 1, Figure 3, Figure 5 and Figure 7 is ⁇ 0.2° ⁇ 0.5°, preferably ⁇ 0.2° ⁇ 0.3°, most preferably ⁇ 0.2° ;
  • the error described in the present invention refers to the X-ray powder diffraction of different batches of samples caused by experimental error factors such as the difference in the level of sample preparation technology, the standardization of operation, the thickness of sample sample preparation, and the use of different sample racks during the experiment.
  • the position of the diffraction peak in the spectrum has a certain degree of overall drift. For those skilled in the art, the overall drift of the diffraction peak position due to experimental errors is predictable.
  • the following tables 5, 6, 7 and 8 are four different The X-ray characteristic diffraction peak data table of the batch crystal form B, and its X-ray powder diffraction patterns are basically shown in Figures 9, 10, 11 and 12:
  • the positions of the corresponding diffraction peaks have drifted to varying degrees.
  • the corresponding 2 ⁇ in 8 are 3.901° (the degree of drift is 0.282°), 4.054° (the degree of drift is 0.129°), 3.973° (the degree of drift is 0.210°), and 4.039° (the degree of drift is 0.144°); or for example
  • the 2 ⁇ is the peak of 16.789°
  • its corresponding 2 ⁇ in Tables 5, 6, 7, and 8 are 16.512° (the degree of drift is 0.277°), 16.634° (the degree of drift is 0.155°), and 16.535° (the degree of drift is 0.155°).
  • the degree is 0.254 °), 16.612 ° (the degree of drift is 0.177 ° ); or for example, the peak of 2 ⁇ in Table 2 is 22.182 °, and its corresponding 2 ⁇ in Table 5, 6, 7, and 8 is respectively 21.915 ° (the degree of drift is 0.267°), 22.020° (the degree of drift is 0.162°), 21.933° (the degree of drift is 0.249°), 22.021° (the degree of drift is 0.161°);
  • the crystal form is an anhydrate.
  • the crystal form is a hydrate, and the number of water is 0.2-3, preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3, more preferably 0.5, 1, 2 or 3 .
  • the preparation method of the crystal form of the above-mentioned compound specifically includes the following steps:
  • Described good solvent is selected from methanol, acetone, ethyl formate, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, One of n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol, 2-butanone or 3-pentanone or Multiple; preferably one or more of ethyl formate, isopropanol or absolute ethanol;
  • the anti-solvent is selected from one or more of heptane, cyclohexane, n-hexane, n-pentane, water, methyl tert-butyl ether, toluene or isopropyl ether; preferably ethyl acetate, methyl One or more of tert-butyl ether or cyclohexane.
  • the preparation method of the crystal form of the above-mentioned compound specifically includes the following steps:
  • Described poor solvent is selected from one or more in heptane, cyclohexane, n-hexane, n-pentane, water, methyl tert-butyl ether, toluene or isopropyl ether; Preferred ethyl acetate, methyl One or more of tert-butyl ether or cyclohexane.
  • the object of the present invention is also to provide a pharmaceutical composition, which contains a therapeutically effective amount of any crystal form of the above compound, and one or more pharmaceutically acceptable carriers, diluents or excipients.
  • the object of the present invention is also to provide the application of the crystal form of any of the above compounds or the above pharmaceutical composition in the preparation of PD-1/PD-L1 inhibitor drugs.
  • the object of the present invention is also to provide the crystal form of any of the above compounds or the application of the above pharmaceutical composition in the preparation of medicines for treating diseases selected from cancer, infectious diseases, and autoimmune diseases;
  • the cancer is selected from skin cancer , lung cancer, urological tumor, blood tumor, breast cancer, glioma, digestive system tumor, reproductive system tumor, lymphoma, nervous system tumor, brain tumor, head and neck cancer;
  • the infectious disease is selected from bacterial infection, viral infection;
  • the autoimmune disease is selected from organ-specific autoimmune disease, systemic autoimmune disease, wherein, the organ-specific autoimmune disease includes chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent Diabetes, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, pulmonary hemorrhage nephritic syndrome, primary biliary cirrhosis, multiple sclerosis, acute idiopathic polyneuritis, the Common systemic autoimmune diseases include r
  • Fig. 1 is an XRPD diagram of free base crystal form A.
  • Fig. 2 is a DSC diagram of free base crystal form A.
  • Figure 3 is an XRPD representation of free base Form B.
  • Fig. 4 is a DSC diagram of free base crystal form B.
  • Figure 5 is an XRPD representation of free base Form C.
  • Figure 6 is a DSC diagram of free base Form C.
  • Figure 7 is an XRPD representation of free base Form D.
  • Fig. 8 is a DSC diagram of free base crystal form D.
  • Fig. 9 is an XRPD diagram of batch 1 of free base crystal form B.
  • Fig. 10 is an XRPD diagram of batch 2 of free-base crystal form B.
  • Fig. 11 is an XRPD diagram of batch 3 of the free-base crystal form B.
  • Fig. 12 is an XRPD diagram of batch 4 of the free base crystal form B.
  • alkyl refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably 1 to 6 carbon atoms An alkyl group, most preferably an alkyl group of 1 to 3 carbon atoms.
  • Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1 ,2-Dimethylpropyl, 2,2-Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2- Methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3 -Dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2 -Methylhexyl, 3-methylhexyl, 4-methylhe
  • lower alkyl groups containing 1 to 6 carbon atoms include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl Base, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl Dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl group, 2,3-dimethylbutyl group, etc.
  • Alkyl groups may be substituted or unsubstituted, and when substituted, substituents may be substituted at any available point of attachment, said substituents being preferably one or more of the following groups independently selected from alkyl radical, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkane Oxygen, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate, preferably methyl, ethyl, isopropyl, tert-butyl, haloalkyl in the present invention , deuterated alkyl, alkoxy substituted alkyl and hydroxy substituted alkyl.
  • alkylene means that one hydrogen atom of the alkyl group is further substituted, for example: "methylene” means -CH 2 -, "ethylene” means -(CH 2 ) 2 -, “propylene” refers to -(CH 2 ) 3 -, “butylene” refers to -(CH 2 ) 4 -, and the like.
  • alkenyl means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, for example vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3- -butenyl etc.
  • Alkenyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycle Alkylthio.
  • cycloalkyl refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing 3 to 20 carbon atoms, preferably containing 3 to 10 carbon atoms, more preferably containing 3 to 8 carbon atoms, more preferably 3 to 6 carbon atoms.
  • Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene group, cyclooctyl group, etc.; polycyclic cycloalkyl group includes spiro ring, fused ring and bridged ring cycloalkyl group, preferably cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl group and cycloheptyl group.
  • the cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring where the ring bonded to the parent structure is a cycloalkyl, non-limiting examples include indanyl, tetrahydronaphthalene base, benzocycloheptyl, etc.
  • Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, oxo, carboxyl or carboxylate.
  • heterocyclyl refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (where m is an integer from 0 to 2), but excluding ring portions of -OO-, -OS- or -SS-, the remaining ring atoms being carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably contains 3 to 10 ring atoms; further preferably contains 3 to 8 ring atoms.
  • Non-limiting examples of monocyclic heterocyclyl groups include oxazinone, pyrazinone, pyridinonyl, pyrrolidinyl, tetrahydropyrrolyl, tetrahydropyrrolidinyl, azetidinyl, oxa Cyclobutanyl, oxanyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuryl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperyl Pyridonyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, tetrahydropyranyl and pyranyl, etc.; preferably oxazinone, pyrazinone, pyridone, Pyrrolidinyl, tetrahydropyrrolyl, te
  • Polycyclic heterocyclic groups include spiro rings, fused rings and bridged ring heterocyclic groups; the spiro rings, condensed rings and bridged ring heterocyclic groups involved are optionally connected to other groups through single bonds, or through rings Any two or more atoms on the ring are further linked with other cycloalkyl, heterocyclyl, aryl and heteroaryl groups.
  • the heterocyclyl ring may be fused to an aryl, heteroaryl, or cycloalkyl ring where the ring bonded to the parent structure is a heterocyclyl, non-limiting examples of which include:
  • Heterocyclic groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alk Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, oxo, carboxyl or carboxylate.
  • aryl refers to a 6 to 14 membered all-carbon monocyclic or fused polycyclic (that is, rings sharing adjacent pairs of carbon atoms) group having a conjugated ⁇ -electron system, preferably 6 to 10 membered, more preferably 6 to 8 membered, such as phenyl and naphthyl. Phenyl is more preferred.
  • the aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, where the ring bonded to the parent structure is an aryl ring, non-limiting examples of which include:
  • Aryl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycle Alkylthio, carboxyl or carboxylate.
  • heteroaryl refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen.
  • Heteroaryl is preferably 5 to 10 membered, more preferably 5 to 8 membered, most preferably 5 or 6 membered, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, Triazolyl, tetrazolyl, pyridyl, pyrimidyl, thiadiazole, pyridazinyl or pyrazinyl, etc.; preferably triazolyl, thienyl, thiazolyl, pyridyl, imidazolyl, pyrazolyl, pyridazine Base, pyrazinyl or pyrimidinyl; more preferably pyridyl, imidazolyl,
  • Heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, carboxyl or carboxylate.
  • alkoxy refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as above, preferably alkyl containing 1 to 8 carbon atoms, more preferably An alkyl group of 1 to 6 carbon atoms, most preferably an alkyl group of 1 to 3 carbon atoms.
  • alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy.
  • Alkoxy may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkoxy Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, carboxyl or carboxylate.
  • the substituent is preferably one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkoxy Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl,
  • Haloalkyl means an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
  • Haloalkoxy means an alkoxy group substituted with one or more halogens, wherein alkoxy group is as defined above.
  • Hydroalkyl means an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.
  • Haloalkyl means an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
  • Haloalkoxy means an alkoxy group substituted with one or more halogens, wherein alkoxy group is as defined above.
  • Hydroalkyl means an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.
  • Haldroxy means an -OH group.
  • Halogen means fluorine, chlorine, bromine or iodine.
  • Amino refers to -NH2 .
  • Cyano refers to -CN.
  • Niro refers to -NO2 .
  • Carboxy refers to -C(O)OH.
  • THF tetrahydrofuran
  • EtOAc means ethyl acetate
  • MeOH means methanol
  • DMF N,N-dimethylformamide
  • TFA trifluoroacetic acid
  • MeCN refers to acetonitrile
  • DMA refers to N,N-dimethylacetamide.
  • Et2O means diethyl ether
  • DCE 1,2 dichloroethane
  • DIPEA N,N-diisopropylethylamine
  • NBS N-bromosuccinimide
  • NIS N-iodosuccinimide
  • Cbz-Cl refers to benzyl chloroformate
  • Pd 2 (dba) 3 refers to tris(dibenzylideneacetone)dipalladium.
  • Dppf refers to 1,1'-bisdiphenylphosphinoferrocene.
  • HATU refers to 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate.
  • KHMDS refers to potassium hexamethyldisilazide
  • LiHMDS refers to lithium bistrimethylsilylamide.
  • MeLi means methyllithium
  • n-BuLi refers to n-butyllithium
  • NaBH(OAc) 3 refers to sodium triacetoxyborohydride.
  • t-BuONO refers to t-butyl nitrite.
  • EA means ethyl acetate
  • PE petroleum ether
  • DCM dichloromethane
  • STAB sodium triacetoxyborohydride
  • Pd(dcypf)Cl 2 refers to dichloro[1,1'-bis(dicyclohexylphosphino)ferrocene]palladium.
  • X is selected from A, B, or C
  • X is selected from A, B, and C
  • X is A, B, or C
  • X is A, B, and C
  • the hydrogen atoms described in the present invention can be replaced by its isotope deuterium, and any hydrogen atom in the example compounds involved in the present invention can also be replaced by a deuterium atom.
  • Optional or “optionally” means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur.
  • a heterocyclic group optionally substituted with an alkyl group means that an alkyl group may but need not be present, and the description includes cases where the heterocycle group is substituted with an alkyl group and cases where the heterocycle group is not substituted with an alkyl group .
  • Substituted means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms are independently substituted by the corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions and that a person skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when bonded to a carbon atom with an unsaturated (eg, ethylenic) bond.
  • “Pharmaceutical composition” means a mixture containing one or more compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, and other components such as a physiologically/pharmaceutically acceptable carrier and excipients.
  • the purpose of the pharmaceutical composition is to promote the administration to the organism, facilitate the absorption of the active ingredient and thus exert biological activity.
  • the structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) or/and liquid chromatography-mass chromatography (LC-MS). NMR chemical shifts ( ⁇ ) are given in parts per million (ppm).
  • the determination of NMR is to use Bruker AVANCE-400 nuclear magnetic apparatus, and the determination solvent is deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated methanol (CD 3 OD) and deuterated chloroform (CDCl 3 ), and the internal standard is four Methylsilane (TMS).
  • Agilent 1200 Infinity Series mass spectrometer was used for liquid chromatography-mass chromatography LC-MS determination.
  • the determination of HPLC used Agilent 1200DAD high pressure liquid chromatography (Sunfire C18 150 ⁇ 4.6mm column) and Waters 2695-2996 high pressure liquid chromatography (Gimini C18 150 ⁇ 4.6mm column).
  • Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates are used for thin-layer chromatography silica gel plates.
  • the specifications used for TLC are 0.15mm-0.20mm, and the specifications used for thin-layer chromatography separation and purification products are 0.4mm-0.5mm.
  • Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.
  • the starting materials in the examples of the present invention are known and commercially available, or can be synthesized using or following methods known in the art.
  • the first step the preparation of 6-(3-bromo-2-chlorophenyl)-2-methoxynicotinaldehyde
  • 6-(3-Bromo-2-chlorophenyl)-2-methoxynicotinaldehyde 600mg, 1.83mmol
  • pinacol bis-boronate 607mg, 2.39mol
  • the fourth step N-(2,2'-dichloro-3'-(6-methoxy-5-((oxetan-3-ylamino)methyl)pyridin-2-yl)-[1 ,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide preparation of
  • reaction solution was quenched with saturated aqueous sodium bicarbonate (10 mL), extracted with DCM (20 mL), the organic layer was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (259 mg, 51%).
  • the first step tert-butyl ((1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H- Imidazo[4,5-c]pyridine-2-carbobalylamino ⁇ oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridine-3
  • -yl methyl
  • azetidin-3-yl) methyl carbamate
  • the third step N-(3'-(5-((3-(acetylaminomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]
  • reaction solution was diluted with DCM (10 mL), washed with saturated aqueous sodium bicarbonate (5 mL), the organic layer was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (18.6 mg, 76%).
  • reaction solution was diluted with DCM (10 mL), washed with saturated aqueous sodium bicarbonate (5 mL), the organic layer was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (28.3 mg, 36%).
  • the first step tert-butyl 6-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxalylamino ⁇ oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl ) methyl)-2,6-diazaspiro [3.3] the preparation of heptane-2-carboxylate
  • reaction solution was diluted with DCM (20 mL), washed with saturated aqueous sodium bicarbonate (10 mL), the organic layer was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound as a brown oil (125 mg, 48%).
  • the third step N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.3]heptane-2-yl)methyl) -6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H- Preparation of imidazo[4,5-c]pyridine-2-carboxamide
  • Test Example 1 Determination of the binding inhibitory effect of the compound of the present invention on human PD-1/PD-L1
  • the purpose of this test case is to measure the activity of the compound on human PD-1/PD-L1 binding inhibition.
  • a centrifuge (5810R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, a microplate reader was purchased from BioTek, USA, and the model was H1MFD full-function microplate reader.
  • Binding Domain diluent buffer was purchased from Cisbio Company, the article number was 62DLBDDF; MAb Anti-6HIS Eu cryptate Gold was purchased from Cisbio Company, the article number was 61HI2KLA; PAb Anti Human IgG-XL665 was purchased from Cisbio Company, the article number was 61HFCXLB; PD-L1 -His protein was purchased from Abcam Company, the article number is ab167713; PD-1-Fc protein was purchased from R&D Company, the article number was 1086-PD; Detection buffer was purchased from Cisbio Company, the article number was 62SDBRDD; 384-well plates were purchased from PerkinElmer Company, the article number was 6007290.
  • TR-FRET time-resolved fluorescence resonance energy transfer
  • This experiment was carried out in a 384-well plate, the total reaction system was 20 ⁇ L, and the compound was diluted into different concentrations (10 ⁇ M or 1 ⁇ M starting, 3-fold dilution, 11 doses) using the experimental buffer Binding Domain diluent buffer (Cisbio#62DLBDDF) , Dilute PD-L1-His protein (19-238 amino acids) (Abcam#ab167713) to 10nM with experimental buffer, and dilute PD-1-Fc protein (25-167 amino acids) (R&D#1086-PD) with experimental buffer
  • the solution was diluted to 20nM, and the compound, PD-L1 protein, and PD-1 protein were added to a 384-well plate, with a total volume of 10 ⁇ L, placed in a centrifuge at 1000rpm for 1 minute to mix well, incubated at room temperature for 60 minutes, and added to each well.
  • the compound concentration and inhibition rate were used for nonlinear regression fitting using Graphpad Prism software to obtain the IC 50 value, as shown in Table 9 below.
  • Test example 2 Determination of endocytic activity of PD-L1 protein on tumor cell surface induced by compounds of the present invention
  • the purpose of this test case is to test the activity of the compound in inducing endocytosis of PD-L1 on the surface of tumor cells.
  • a centrifuge (5702R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, and a flow cytometer was purchased from Beckman Coulter, model DxFlex.
  • PE Mouse Anti-Human CD274 antibody was purchased from BD Pharmingen Company, the article number is 557924; BSA was purchased from Sigma Company, the article number was B2064-100G; PBS was purchased from Gibco Company, the article number was 10010049; 24-well plates were purchased from Corning Company, The item number is 3526.
  • the mouse colon cancer cells MC38-hPDL1 with high expression of hPD-L1 were collected, adjusted to a suitable density, spread on a 24-well plate, and placed in a 37°C, 5% CO 2 incubator to adhere to the wall overnight.
  • the compound was prepared into different concentrations in the medium and added to the 24-well plate, and the solvent control well was set, and incubated in a 37°C, 5% CO2 incubator for 16 hours, then the 24-well plate was taken out, and different treatments in the plate were collected.
  • MC38-hPDL1 cells were washed once with FACS buffer (PBS containing 0.5% BSA), and the cells were prepared to a suitable density with FACS buffer, and PE Mouse Anti-Human CD274 antibody (BD Pharmingen#557924) was added, and shaken at room temperature Incubate in the dark for 30 minutes, wash the cells twice with FACS buffer, resuspend in 100 ⁇ L PBS, use flow cytometry to detect the fluorescent signal on the cell surface, and set the isotype control as a negative control.
  • FACS buffer PBS containing 0.5% BSA
  • PE Mouse Anti-Human CD274 antibody BD Pharmingen#557924
  • the endocytosis rate of the compound was calculated using the fluorescence signals of different treatment groups, and the concentration of the compound and the endocytosis rate were used for nonlinear regression fitting using GraphPad Prism software to obtain the IC 50 value, as shown in Table 10.
  • the compounds shown in the present invention can well induce the endocytosis of PD-L1 on the surface of tumor cells.
  • Test example 3 Inhibitory effect of the compound of the present invention on CHO-PDL1/Jurkat-PD1 cell pathway
  • the purpose of this test case is to test the inhibitory effect of the compound on the CHO-PDL1/Jurkat-PD1 cellular pathway.
  • a centrifuge (5810R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, a microplate reader was purchased from BioTek, USA, and the model was H1MFD full-function microplate reader.
  • CHO-PDL1 cells and NFAT-luc2/PD1 Jurkat cells were purchased from Promega Company, product number J1252; RPMI 1640 was purchased from Gibco Company, product number 22400089; FBS was purchased from Gibco Company, product number 10091148; One-Glo reagent was purchased from The 96-well plate was purchased from Promega Company, the product number is E6120; the 96-well plate was purchased from Corning Company, the product number is 3610.
  • Cultivate CHO-PDL1 cells (Promega, #J1252) to a suitable cell density, digest and resuspend the cell suspension with complete medium to a suitable density, spread 100 ⁇ L per well on a 96-well plate (Corning, #3610), put 37°C, 5% CO 2 incubator adherent culture overnight, use analysis medium (RPMI 1640+2% FBS, RPMI 1640 product number is Gibco, #22400089, FBS product number is Gibco, #10091148) to prepare compound solutions with different concentrations, NFAT-luc2/PD1 Jurkat cells (Promega, #J1252) were collected and resuspended to an appropriate cell density using assay medium.
  • RPMI 1640+2% FBS RPMI 1640 product number is Gibco, #22400089, FBS product number is Gibco, #10091148
  • the EC 50 value was fitted according to the compound concentration and the luminescent signal value, and the maximum induction factor of each compound relative to the signal value of the untreated group was calculated, as shown in Table 11.
  • Test Example 4 Determination of the Combination of Invention Compounds to Improve PD-L1 Protein Stability (Melting Temperature)
  • the purpose of the experiment To test the ability of the compound to improve the stability of PD-L1 protein and increase the melting temperature of the protein.
  • Quantitative PCR instrument (Quantstudio6 Flex) was purchased from Life Company, and the pipette was purchased from Eppendorf or Rainin Company.
  • Protein Thermal Shift TM Dye Kit was purchased from Thermofisher Company, product number 4461146; PD-L1 protein was purchased from Acro Biosystems Company, product number was PD1-H5229; HEPES, 1M Buffer Solution was purchased from Thermofisher Company, product number 15630080; NaCl was purchased from From Sinopharm Chemical Reagent Co., Ltd., the product number is 10019318.
  • the compounds shown in the present invention have the ability to improve the stability of PD-L1 protein in the test of increasing the melting temperature of PD-L1 protein.
  • Test example 5 The in vitro cytotoxic effect of the compound of the present invention on NFAT-luc2/PD1 Jurkat cells and MC38-hPDL1 cells
  • a centrifuge (5810R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, a microplate reader was purchased from BioTek, USA, and the model was H1MFD full-function microplate reader.
  • RPMI 1640 was purchased from Gibco Company, the product number is 22400089; FBS was purchased from Gibco Company, the product number was 10091148; PBS was purchased from Gibco Company, the product number was 10010049; Cell Titer-Glo was purchased from Promega Company, the product number was G7573; cell culture plate Purchased from Corning Company, Cat. No. 3610.
  • the cells When the cells are cultured to a suitable degree of confluence, collect the cells, use the complete medium to adjust the cells to a suitable cell concentration, spread the cell suspension on a 96-well plate, 90 ⁇ L per well, and place it in a 37°C, 5% CO 2 incubator Adhere to the wall overnight, use DMSO and medium to prepare compound solutions of different concentrations, set up vehicle control, add the compound solution to a 96-well plate, 10 ⁇ L per well, put it in a 37°C, 5% CO 2 incubator and continue to cultivate for 72 hours , add CellTiter-Glo solution, shake and mix well, incubate in the dark for 10 minutes, and read with BioTek Synergy H1 microplate reader.
  • the compounds shown in the present invention have weak toxic effects on NFAT-luc2/PD1 Jurkat and MC38-hPDL1 cells.
  • Test example 6 Balb/C mouse pharmacokinetics is measured
  • mice Taking Balb/C mice as the experimental animals, the pharmacokinetics of the following compound examples were orally administered at a dose of 5 mg/kg in mice in vivo.
  • Embodiment of the present invention self-made.
  • HPMC 1% Tween 80
  • Balb/C mice male; p.o. after fasting overnight, the dose is 5mg/kg, and the administration volume is 10mL/kg.
  • liquid A is 0.1% formic acid aqueous solution
  • liquid B is acetonitrile
  • Test example 7 Pharmacokinetic determination of tumor-bearing nude mouse plasma and tumor
  • Embodiment of the present invention self-made.
  • HPMC 1% Tween 80
  • Tumor-bearing nude mice female; p.o. after fasting overnight, the dose is 30mg/kg, and the administration volume is 10mL/kg.
  • Tumor tissue was homogenized by adding methanol water at a ratio of 1:3 and centrifuged to obtain the supernatant. Take 40 ⁇ L of the tumor homogenate supernatant and plasma samples and add 160 ⁇ L of acetonitrile for precipitation. After mixing, centrifuge at 3500 ⁇ g for 5-20 minutes.
  • liquid A is 0.1% formic acid aqueous solution
  • liquid B is acetonitrile
  • Test example 8 plasma protein binding rate experiment
  • the purpose of this experimental method is to detect the plasma protein binding of the compounds of the examples in plasma.
  • Ultra-high performance liquid phase tandem mass spectrometry coupled instrument refrigerated centrifuge, vortex instrument, electric heating constant temperature oscillating water tank, pipette, continuous liquid adder, 96-well plate, tissue homogenizer (used for tissue sample analysis), Add internal standard acetonitrile solution, blank matrix (plasma, urine or tissue homogenate, etc.)
  • Compound working solution preparation dilute the stock solution with DMSO to a final concentration of 1 mM.
  • Washing gradient 0.2 ⁇ 1.6min 5%A to 95%A, 3.0 ⁇ 3.1min 95%A to 5%A;
  • Ion source electrospray ionization source (ESI);
  • Drying gas N 2 , temperature 500°C;
  • Electrospray voltage 5000V
  • Detection method positive ion detection
  • Scanning method select the reaction monitoring (MRM) method.
  • the compound of the embodiment of the present invention shows a high plasma protein binding rate.
  • Vernier caliper 500-196, Mitutoyo, Japan
  • Fetal bovine serum (FBS) (10099-141, Gibco);
  • PBS Phosphate buffered saline
  • the compound of the embodiment of the present invention is self-made.
  • MC38-hPDL1 cells Take out MC38-hPDL1 cells from the cell bank, add DMEM medium (DMEM+10% FBS) after recovery, and place them in a CO 2 incubator for culture (incubator temperature is 37°C, CO 2 concentration is 5%). When the number expanded to the required number for inoculation in vivo, MC38-hPDL1 cells were harvested. Count with an automatic cell counter, and resuspend the cells with PBS according to the counting results to make a cell suspension (density 1 ⁇ 10 6 /mL), and place in an ice box until use.
  • DMEM medium DMEM+10% FBS
  • mice Use 6-8 week old female C57 mice weighing approximately 18-22 g. Mice were raised in SPF grade animal room, and raised in single cages, with 4-5 mice per cage. All cages, litter and water were sterilized before use, and all animals had free access to food and water. Before the experiment started, the C57 mice were marked with a disposable ear tag for rats and rats, and the skin of the inoculation site was disinfected with 75% alcohol before inoculation, and each mouse was subcutaneously inoculated with 0.1 mL (containing 1*10 5 cells) of MC38- hPDL1 cells. When the tumor volume reached 40-180mm 3 , group administration began, with 8 rats in each group. Each test compound was orally administered twice a day for 14 days. Tumor volume was measured and mouse body weight was weighed twice a week, and tumor TGI (%) was calculated.
  • Tumor volume (mm 3 )
  • TGI (%) [(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group))/(average tumor volume at the end of treatment of the solvent control group - Average tumor volume at the beginning of treatment in solvent control group)] ⁇ 100%;
  • TGI (%) [1-(average tumor volume at the end of administration of a treatment group-average tumor volume at the beginning of administration of this treatment group)/average tumor volume at the beginning of administration of this treatment group] ⁇ 100%.
  • Example 4 Compound SD rats repeated gavage toxicity test for 14 days
  • the purpose of this study is to investigate the possible toxic reaction of the compound of Example 4 given to SD rats after repeated gavage for 14 days.
  • Test article N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine-2 -yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazole A[4,5-c]pyridine-2-carboxamide (compound of Example 4)
  • the average exposure amount (AUC last ) of the compound of Example 4 in the animal body all increases with dosage; There was no significant gender difference in the exposure of the compound of Example 4 in animals. After 14 days of repeated administration, compared with the first administration, there was no significant accumulation of the compound of Example 4 in the animals of each dose group.
  • FIB Neut and FIB increased in 100mg/kg dose group; FIB increased in female animals in 15 and 50mg/kg dose groups.
  • the compound of Example 4 was administered to SD rats repeatedly by intragastric administration at doses of 15, 50, and 100 mg/kg, once a day, for 14 consecutive days, and the maximum tolerated dose (MTD) was 100 mg/kg.
  • the obtained solid detection XRPD is Form A. After detection and analysis, it has an XRPD pattern as shown in FIG. 1 and a DSC pattern as shown in FIG. 2 .
  • the obtained solid detection XRPD is crystal form B. After detection and analysis, it has an XRPD pattern as shown in FIG. 3 and a DSC pattern as shown in FIG. 4 .
  • a relatively stable crystal form of the compound of Example 4 was found through the crystal form beating experiment stability investigation experiment.
  • the free base crystal form B has good stability under high temperature and high humidity conditions, and the maximum single-heterogeneity increase is less than 0.1%.
  • the hygroscopicity of the free base crystal form B of the compound of Example 4 was investigated under different relative humidity conditions to provide a basis for the screening and storage of the compound crystal form.
  • the free base crystal form B of the compound is placed in saturated water vapor with different relative humidity, so that the compound and the water vapor reach a dynamic equilibrium, and the percentage of the weight gain of the compound after the equilibrium is calculated.
  • the free base crystal form B has a hygroscopic weight gain of about 0.815%, which is slightly hygroscopic. After one cycle of moisture absorption and desorption under the condition of 0-95% relative humidity, the XRPD spectrum of the free base crystal form B does not change, that is, the crystal form B is not transformed.
  • thermodynamic solubility of the compound at 37°C was determined by HPLC and external standard method.
  • Free base crystal form B has good solubility in the pH range and environment of the entire gastrointestinal tract.

Abstract

A crystal form of a compound of general formula (I), a preparation method therefor, a pharmaceutical composition containing the therapeutically effective amount of the crystal form, and the use thereof as a PD-1/PD-L1 inhibitor in the treatment of cancers, infectious diseases and autoimmune diseases.

Description

一种联苯类衍生物抑制剂的晶型及其制备方法Crystal form of biphenyl derivative inhibitor and preparation method thereof 技术领域technical field
本发明属于药物化学技术领域，具体涉及一种联苯类衍生物抑制剂的晶型及制备方法和应用。The invention belongs to the technical field of medicinal chemistry, and in particular relates to a crystal form of a biphenyl derivative inhibitor, a preparation method and application thereof.
背景技术Background technique
免疫系统在控制癌症等多种疾病中发挥重要的作用。然而，肿瘤细胞会通过各种途径逃避免疫攻击或者抑制免疫系统的激活。阻断免疫抑制检查点的信号传递，如程序性细胞死亡受体1(programmed cell death protein 1,PD-1)被证明是一种有潜力的治疗方式。The immune system plays an important role in controlling many diseases, including cancer. However, tumor cells evade immune attack or suppress the activation of the immune system in various ways. Blocking signaling at immune-suppressive checkpoints, such as programmed cell death protein 1 (PD-1), has proven to be a potential therapeutic modality.
PD-1为CD28超家族成员，是免疫细胞特别是细胞毒T细胞表面的免疫抑制性受体。PD-1具有两个配体PD-L1和PD-L2，其中PD-L1，细胞程序性死亡受体-配体1，在多种细胞中均有表达，如巨噬细胞和树突状细胞，并在肿瘤细胞上普遍高表达。PD-L1通过与PD-1结合发挥免疫抑制作用并使肿瘤细胞逃避T细胞的杀伤，抑制T细胞的激活和相应细胞因子的产生，减弱感染性免疫和肿瘤免疫，促进感染性疾病以及肿瘤的进展。使用PD-L1抑制剂如抗体或小分子抑制剂可解除PD-L1的免疫抑制作用，促进肿瘤被免疫清除，从而达到治疗肿瘤的效果。PD-1, a member of the CD28 superfamily, is an immunosuppressive receptor on the surface of immune cells, especially cytotoxic T cells. PD-1 has two ligands PD-L1 and PD-L2, among which PD-L1, programmed cell death receptor-ligand 1, is expressed in a variety of cells, such as macrophages and dendritic cells , and generally highly expressed on tumor cells. PD-L1 plays an immunosuppressive role by combining with PD-1 and makes tumor cells evade the killing of T cells, inhibits the activation of T cells and the production of corresponding cytokines, weakens infectious immunity and tumor immunity, and promotes the development of infectious diseases and tumors. progress. The use of PD-L1 inhibitors such as antibodies or small molecule inhibitors can relieve the immunosuppressive effect of PD-L1 and promote the immune clearance of tumors, thereby achieving the effect of treating tumors.
PD-1/PD-L1是近年来肿瘤免疫疗法研究的热点，其单抗药物应答之广度、深度和持久性均十分罕见，目前临床上已有多个PD-1/PD-L1单抗药物上市，并取得的巨大的成功。PD-L1抑制剂可用于治疗包括非小细胞肺癌、肝癌、胃癌、肠癌、肾癌等几乎所有重大癌症，具有巨大的临床应用价值。PD-1/PD-L1 is a hot spot in the research of tumor immunotherapy in recent years. The breadth, depth and durability of the monoclonal antibody drug response are very rare. At present, there are many PD-1/PD-L1 monoclonal antibody drugs in clinical practice. marketed and achieved great success. PD-L1 inhibitors can be used to treat almost all major cancers, including non-small cell lung cancer, liver cancer, gastric cancer, intestinal cancer, kidney cancer, etc., and have great clinical application value.
PD-L1抑制剂从大分子到小分子正成为一种新的研发趋势和热点，小分子抑制剂从给药方式到生产成本都具有许多天然优势，具有取代抗体大分子的潜力，目前包括BMS和Incyte等公司在内的国外药企均在积极进行PD-L1小分子抑制剂的开发。PD-L1 inhibitors from macromolecules to small molecules are becoming a new research and development trend and hotspot. Small molecule inhibitors have many natural advantages in terms of administration methods and production costs, and have the potential to replace antibody macromolecules, including BMS at present. Foreign pharmaceutical companies including Incyte and other companies are actively developing PD-L1 small molecule inhibitors.
BMS公司开发的口服小分子抑制剂目前处于临床前研究阶段，已连续发表数篇专利，Incyte公司开发的小分子抑制剂INCB086550已处于一期临床研究阶段，Aurigene/Curis公司开发的小分子抑制剂CA-170已处于临床二期研究阶段。The oral small-molecule inhibitor developed by BMS is currently in the preclinical research stage, and several patents have been published consecutively. The small-molecule inhibitor INCB086550 developed by Incyte is in the first phase of clinical research. The small-molecule inhibitor developed by Aurigene/Curis CA-170 is in Phase II clinical research.
国际申请WO2015034820、WO2015160641、WO2014151634、WO2017066227、WO2017070089、WO2017106634、WO2017112730、WO2017192961、WO2017222976、WO2018013789、WO2018044783、WO2018119224、WO2018119236、WO2018119263、WO2018119266和WO2018119286等多篇专利报道了PD-1或PD-L1小分子化合物抑制剂。此外，国际申请WO2014151634、WO2011161699、WO2012168944、WO2013132317、WO2013144704、 WO2015033299、WO2015033301、WO2015033303和WO2015036927报道了大环类和肽类化合物PD-1或PD-L1抑制剂。然而，对PD-1/PD-L1通路更有效、更好的药代特性和成药性的PD-L1小分子抑制剂仍然存在极大需求。国际申请WO2015034820、WO2015160641、WO2014151634、WO2017066227、WO2017070089、WO2017106634、WO2017112730、WO2017192961、WO2017222976、WO2018013789、WO2018044783、WO2018119224、WO2018119236、WO2018119263、WO2018119266和WO2018119286等多篇专利报道了PD-1或PD-L1小分子化合物抑制agent. In addition, international applications WO2014151634, WO2011161699, WO2012168944, WO2013132317, WO2013144704, WO2015033299, WO2015033301, WO2015033303 and WO2015036927 reported macrocyclic and peptide-based PD1-1 inhibitors. However, there is still a great demand for PD-L1 small molecule inhibitors that are more effective, better pharmacokinetic and druggable for the PD-1/PD-L1 pathway.
PD-L1小分子抑制剂作为药物在医药行业具有良好的应用前景，首先，PD-L1小分子抑制剂可口服给药，具有比抗体药物静脉给药顺应性更强的优势，同时可避免抗体在体内长期驻留造成的结肠炎等严重副作用。其次，PD-L1小分子抑制剂具有结合PD-L1并使之内吞的独特作用机制，在临床上可能展示出和抗体不同的功效。最后，PD-L1小分子抑制剂生产和质量控制成本更低，具有远低于大分子药物成本的价格优势，且和PD-L1抗体抑制剂一样可应用于多种重大肿瘤，具有巨大的市场潜力。Small molecule inhibitors of PD-L1 have good application prospects as drugs in the pharmaceutical industry. First, small molecule inhibitors of PD-L1 can be administered orally, which has the advantage of stronger compliance than intravenous administration of antibody drugs, and can avoid antibody Serious side effects such as colitis caused by long-term residence in the body. Secondly, small molecule inhibitors of PD-L1 have a unique mechanism of action that binds to PD-L1 and endocytizes it, which may show different efficacy from antibodies in clinical practice. Finally, the production and quality control costs of PD-L1 small molecule inhibitors are lower, and they have a price advantage that is far lower than the cost of macromolecular drugs. Like PD-L1 antibody inhibitors, they can be applied to a variety of major tumors and have a huge market potential.
江苏豪森药业集团有限公司的专利申请(申请号：PCT/CN2020/141307)中公开了一系列含联苯类衍生物抑制剂的结构，在后续的研发中，为了使产物易于处理、过滤、干燥，便于储存、产品长期稳定、生物利用度高等，本发明对上述物质的晶型进行了全面的研究，致力于得到最适合的晶型。The patent application of Jiangsu Hansoh Pharmaceutical Group Co., Ltd. (Application No.: PCT/CN2020/141307) disclosed a series of structures containing inhibitors of biphenyl derivatives. In subsequent research and development, in order to make the products easy to handle, filter , drying, easy storage, long-term product stability, high bioavailability, etc., the present invention has carried out comprehensive research on the crystal forms of the above substances, and is committed to obtaining the most suitable crystal forms.
发明内容Contents of the invention
专利申请PCT/CN2020/141307所涉及的所有内容均以引证的方式添加到本发明中。All content involved in the patent application PCT/CN2020/141307 is added to the present invention by reference.
本发明的目的在于提供一种通式(I)所示的化合物的晶型，其结构如下：The object of the present invention is to provide a kind of crystal form of the compound shown in general formula (I), its structure is as follows:
Figure PCTCN2022102195-appb-000001
Figure PCTCN2022102195-appb-000001
其中：in:
L选自键、
Figure PCTCN2022102195-appb-000002
L is selected from the key,
Figure PCTCN2022102195-appb-000002
环A选自4-8元杂环基；优选4元、7元或8元杂环基；Ring A is selected from 4-8 membered heterocyclic groups; preferably 4-membered, 7-membered or 8-membered heterocyclic groups;
更优选如下基团：More preferred are the following groups:
Figure PCTCN2022102195-appb-000003
Figure PCTCN2022102195-appb-000003
R 1选自氢、氘、卤素、羟基、氰基、羧基、醛基、氧代基、C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6羟烷基、C 3-8环烷基、-(CH 2) nR a、-(CH 2) nOR a、-OC(O)R a、-C(O)R a、-C(O)OR a、-C(O)NR aR b、-(CH 2) nNR aC(O)R b或-SO 2R a，其中所述的C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6羟烷基和C 3-8环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-6烷基中的一个或多个取代基所取代； R 1 is selected from hydrogen, deuterium, halogen, hydroxyl, cyano, carboxyl, aldehyde, oxo, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O)R a , -C( O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a , wherein the C 1 -6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl and C 3-8 cycloalkyl, optionally further deuterated , halogen, hydroxyl, cyano or one or more substituents in C 1-6 alkyl;
R a和R b各自独立地选自氢、氘、卤素、氨基、羟基、巯基、氰基、硝基、羧基、C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基或C 3-8环烷基，其中所述的C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基和C 3-8环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-6烷基中的一个或多个取代基所取代； R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, carboxyl, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 Haloalkyl or C 3-8 cycloalkyl, wherein said C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl and C 3-8 cycloalkyl, optionally further One or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-6 alkyl;
n为0、1或2；且n is 0, 1 or 2; and
x为0、1或2。x is 0, 1 or 2.
在本发明的优选方案中，所述化合物的结构如通式(II)所示：In a preferred embodiment of the present invention, the structure of the compound is shown in general formula (II):
Figure PCTCN2022102195-appb-000004
Figure PCTCN2022102195-appb-000004
其中：in:
M为O、-NR 2或-CR 3R 4； M is O, -NR 2 or -CR 3 R 4 ;
R 2选自氢、氘、醛基、C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基、C 1-3烷氧基、C 1-3羟烷基、C 3-6环烷基、-(CH 2) nR a、-(CH 2) nOR a、-OC(O)R a、-C(O)R a、-C(O)OR a、-C(O)NR aR b、-(CH 2) nNR aC(O)R b或-SO 2R a； R 2 is selected from hydrogen, deuterium, aldehyde, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 3-6 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O)R a , -C(O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a ;
R 3和R 4各自独立地选自氢、氘、卤素、羟基、氰基、羧基、醛基、氧代基、C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基、C 1-3烷氧基、C 1-3羟烷基、C 3-6环烷基、-(CH 2) nR a、-(CH 2) nOR a、-OC(O)R a、-C(O)R a、-C(O)OR a、-C(O)NR aR b、-(CH 2) nNR aC(O)R b或-SO 2R a，其中所述的C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基、C 1-3烷氧基、C 1-3羟烷基和C 3-6环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-3烷基中的一个或多个取代基所取代； R 3 and R 4 are each independently selected from hydrogen, deuterium, halogen, hydroxyl, cyano, carboxyl, aldehyde, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 Haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 3-6 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O) R a , -C(O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a , The C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl and C 3-6 cycloalkyl , optionally further substituted by one or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-3 alkyl;
R a和R b各自独立地选自氢、氘、卤素、氨基、羟基、巯基、氰基、硝基、羧基、C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基或C 3-6环烷基，其中所述的C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基和C 3-6环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-3烷基中的一个或多个取代基所取代； R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, carboxyl, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 Haloalkyl or C 3-6 cycloalkyl, wherein said C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl and C 3-6 cycloalkyl, optionally further One or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-3 alkyl;
n为0、1或2。n is 0, 1 or 2.
在本发明的优选方案中，所述化合物的晶型，为以下化合物的晶型：In a preferred embodiment of the present invention, the crystal form of the compound is the crystal form of the following compound:
N-(2,2'-二氯-3'-(6-甲氧基-5-((噁丁环-3-基氨基)甲基)吡啶-2-基)-[1,1'-联苯 基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((oxetan-3-ylamino)methyl)pyridin-2-yl)-[1,1'- Biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((3-氟吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-fluoroazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-基乙酸酯；1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-carbobalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine- 3-yl acetate;
N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
N-(3'-(5-((2-氧杂-6-氮杂螺[3.3]庚烷-6-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((2-Oxa-6-azaspiro[3.3]heptane-6-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]pyridine-2-carboxamide;
N-(3'-(5-((5-氧杂-2-氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((5-oxa-2-azaspiro[3.4]octane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-羰基-2,5-二氮杂螺[3.4]辛烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-carbonyl-2,5-diazaspiro[3.4]octane-2-yl)methyl) Pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5 -c] pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((3-羟基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-hydroxyazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((3-羟基-3-甲基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-hydroxy-3-methylazetidin-1-yl)methyl)-6-methoxypyridin-2-yl) -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 - Formamide;
1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺基)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-羧酸；1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-Carboxamido)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine-3-carboxylic acid;
(S)-1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-2-羧酸；(S)-1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5 -c]pyridine-2-carboxalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl) Azetidine-2-carboxylic acid;
(S)-N-(2,2'-二氯-3'-(5-((2-(羟甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；(S)-N-(2,2'-dichloro-3'-(5-((2-(hydroxymethyl)azetidin-1-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((3-羟基-3-(羟甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-hydroxy-3-(hydroxymethyl)azetidin-1-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide;
N-(3'-(5-((3-乙酰氨基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((3-acetylaminoazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1, 1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
N-(3'-(5-((3-(乙酰氨基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((3-(acetylaminomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 - Formamide;
N-(2,2'-二氯-3'-(5-((3-(氰基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-(cyanomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl) -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 - Formamide;
N-(2,2'-二氯-3'-(5-((3-(二甲基氨基甲酰)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2- 基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-(dimethylcarbamoyl)azetidin-1-yl)methyl)-6-methoxypyridine-2- base)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-formamide;
N-(2,2'-二氯-3'-(6-甲氧基-5-((3-(甲基氨基甲酰)吖丁啶-1-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((3-(methylcarbamoyl)azetidin-1-yl)methyl)pyridin-2-yl )-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine- 2-formamide;
N-(2,2'-二氯-3'-(5-((6-羟基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-hydroxy-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methanol Oxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4 ,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((6-(环丙羰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-(cyclopropylcarbonyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6 -Methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-丙酰-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-propionyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(2,2,2-三氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(2,2,2-trifluoroacetyl)-2,6-diazaspiro[3.3 ]heptane-2-yl)methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7- Tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((6-(2,2-二氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-(2,2-difluoroacetyl)-2,6-diazaspiro[3.3]heptan-2-yl) Methyl)-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro -1H-imidazo[4,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((6-(2-氰基乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-(2-cyanoacetyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H - imidazo[4,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(甲磺酰)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(methylsulfonyl)-2,6-diazaspiro[3.3]heptane-2-yl )methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((6-甲酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-formyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-6-methoxy Basepyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
N-(3'-(5-((7-乙酰基-2,7-二氮杂螺[3.5]壬烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((7-acetyl-2,7-diazaspiro[3.5]nonan-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡 啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；或N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methanol Oxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4 ,5-c]pyridine-2-carboxamide; or
N-(3'-(5-((6-乙酰氨基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺。N-(3'-(5-((6-Acetamido-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ] Pyridine-2-carboxamide.
在本发明的优选方案中，所述晶型为化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型。In a preferred embodiment of the present invention, the crystal form is the compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Crystalline form of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide.
在本发明的优选方案中，提供化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型A～D，其中：In a preferred embodiment of the present invention, the compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6- Methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7- Crystal forms A to D of tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, wherein:
晶型A：Form A:
所述晶型A的X-射线粉末衍射图谱在2θ为7.8±0.2°处具有衍射峰；或者在14.1±0.2°处具有衍射峰；或者在15.3±0.2°处具有衍射峰；或者在19.5±0.2°处具有衍射峰；或者在12.0±0.2°处具有衍射峰；或者在12.4±0.2°处具有衍射峰；或者在13.2±0.2°处具有衍射峰；或者在14.6±0.2°处具有衍射峰；或者在21.2±0.2°处具有衍射峰；或者在22.0±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form A has a diffraction peak at 2θ of 7.8±0.2°; or has a diffraction peak at 14.1±0.2°; or has a diffraction peak at 15.3±0.2°; or has a diffraction peak at 19.5±0.2°; Diffraction peak at 0.2°; or diffraction peak at 12.0±0.2°; or diffraction peak at 12.4±0.2°; or diffraction peak at 13.2±0.2°; or diffraction peak at 14.6±0.2° or have a diffraction peak at 21.2±0.2°; or have a diffraction peak at 22.0±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
或，晶型B：or, Form B:
所述晶型B的X-射线粉末衍射图谱在2θ为4.2±0.2°处具有衍射峰；或者在12.2±0.2°处具有衍射峰；或者在12.6±0.2°处具有衍射峰；或者在16.5±0.2°处具有衍射峰；或者在16.8±0.2°处具有衍射峰；或者在18.4±0.2°处具有衍射峰；或者在18.9±0.2°处具有衍射峰；或者在21.1±0.2°处具有衍射峰；或者在22.2±0.2°处具有衍射峰；或者在22.4±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form B has a diffraction peak at 2θ of 4.2±0.2°; or has a diffraction peak at 12.2±0.2°; or has a diffraction peak at 12.6±0.2°; or has a diffraction peak at 16.5±0.2°; Diffraction peak at 0.2°; or diffraction peak at 16.8±0.2°; or diffraction peak at 18.4±0.2°; or diffraction peak at 18.9±0.2°; or diffraction peak at 21.1±0.2° or have a diffraction peak at 22.2±0.2°; or have a diffraction peak at 22.4±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
或，晶型C：or, Form C:
所述晶型C的X-射线粉末衍射图谱在2θ为11.4±0.2°处具有衍射峰；或者在12.5±0.2°处具有衍射峰；或者在21.1±0.2°处具有衍射峰；或者在23.0±0.2°处具有衍射峰；或者在26.1±0.2°处具有衍射峰；或者在26.6±0.2°处具有衍射峰；或者在13.0±0.2°处具有衍射峰；或者在14.0±0.2°处具有衍射峰；或者在14.7±0.2°处具有衍射峰；或者在15.8±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form C has a diffraction peak at 2θ of 11.4±0.2°; or has a diffraction peak at 12.5±0.2°; or has a diffraction peak at 21.1±0.2°; or has a diffraction peak at 23.0±0.2°; Diffraction peak at 0.2°; or diffraction peak at 26.1±0.2°; or diffraction peak at 26.6±0.2°; or diffraction peak at 13.0±0.2°; or diffraction peak at 14.0±0.2° or have a diffraction peak at 14.7±0.2°; or have a diffraction peak at 15.8±0.2°; preferably include any 2-5 of the above-mentioned diffraction peaks, or 3-5, or 3-6, or 3 -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
或，晶型D：or, Form D:
所述晶型D的X-射线粉末衍射图谱在2θ为9.5±0.2°处具有衍射峰；或者在 10.4±0.2°处具有衍射峰；或者在14.5±0.2°处具有衍射峰；或者在19.9±0.2°处具有衍射峰；或者在20.8±0.2°处具有衍射峰；或者在24.8±0.2°处具有衍射峰；或者在11.7±0.2°处具有衍射峰；或者在13.3±0.2°处具有衍射峰；或者在17.2±0.2°处具有衍射峰；或者在23.8±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处。The X-ray powder diffraction pattern of the crystal form D has a diffraction peak at 2θ of 9.5±0.2°; or has a diffraction peak at 10.4±0.2°; or has a diffraction peak at 14.5±0.2°; or has a diffraction peak at 19.9±0.2°; Diffraction peak at 0.2°; or diffraction peak at 20.8±0.2°; or diffraction peak at 24.8±0.2°; or diffraction peak at 11.7±0.2°; or diffraction peak at 13.3±0.2° or have a diffraction peak at 17.2±0.2°; or have a diffraction peak at 23.8±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places.
在本发明进一步优选的方案中，所述晶型A的X-射线粉末衍射图谱至少包含位于2θ为7.8±0.2°、14.1±0.2°、15.3±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含19.5±0.2°、12.0±0.2°、12.4±0.2°、13.2±0.2°、14.6±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；In a further preferred solution of the present invention, the X-ray powder diffraction pattern of the crystal form A contains at least one or more diffraction peaks at 2θ of 7.8±0.2°, 14.1±0.2°, and 15.3±0.2°, Preferably two of them are included, more preferably three; optionally, at least one of 19.5±0.2°, 12.0±0.2°, 12.4±0.2°, 13.2±0.2°, 14.6±0.2° can be included, preferably 2, 3, 4 or 5 of them;
或，所述晶型B的X-射线粉末衍射图谱至少包含位于2θ为4.2±0.2°、12.2±0.2°、12.6±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含16.5±0.2°、16.8±0.2°、18.4±0.2°、18.9±0.2°、21.1±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；Or, the X-ray powder diffraction pattern of the crystal form B includes at least one or more diffraction peaks located at 2θ of 4.2±0.2°, 12.2±0.2°, and 12.6±0.2°, preferably two of them, and more It preferably contains three; optionally, it can further include at least one of 16.5±0.2°, 16.8±0.2°, 18.4±0.2°, 18.9±0.2°, 21.1±0.2°, preferably 2, 3, 4 or 5;
或，所述晶型C的X-射线粉末衍射图谱至少包含位于2θ为11.4±0.2°、12.5±0.2°、21.1±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含23.0±0.2°、26.1±0.2°、26.6±0.2°、13.0±0.2°、14.0±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；Or, the X-ray powder diffraction pattern of the crystal form C contains at least one or more diffraction peaks located at 2θ of 11.4±0.2°, 12.5±0.2°, and 21.1±0.2°, preferably two of them, and more It preferably contains three; optionally, it can further contain at least one of 23.0±0.2°, 26.1±0.2°, 26.6±0.2°, 13.0±0.2°, 14.0±0.2°, preferably 2, 3, 4 or 5;
或，所述晶型D的X-射线粉末衍射图谱至少包含位于2θ为9.5±0.2°、10.4±0.2°、14.5±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含19.9±0.2°、20.8±0.2°、24.8±0.2°、11.7±0.2°、13.3±0.2°中的至少一条，优选包含其中2条、3条、4条或5条。Or, the X-ray powder diffraction pattern of the crystal form D contains at least one or more diffraction peaks located at 2θ of 9.5±0.2°, 10.4±0.2°, and 14.5±0.2°, preferably two of them, and more Preferably include three; optionally, it may further include at least one of 19.9±0.2°, 20.8±0.2°, 24.8±0.2°, 11.7±0.2°, 13.3±0.2°, preferably including 2, 3, 4 or 5 bars.
在本发明进一步优选的方案中，所述晶型A的X-射线粉末衍射图谱任选还包含位于2θ为21.2±0.2°、22.0±0.2°、22.6±0.2°、25.0±0.2°、28.3±0.2°、16.7±0.2°或24.5±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；In a further preferred solution of the present invention, the X-ray powder diffraction pattern of the crystal form A optionally further comprises One or more diffraction peaks at 0.2°, 16.7±0.2° or 24.5±0.2°; preferably at least any 2-3, or 4-5, or 6-7 of them; more preferably, including Any 2, 3, 4, 5, 6, 7 places;
或，所述晶型B的X-射线粉末衍射图谱任选还包含位于2θ为22.2±0.2°、22.4±0.2°、22.9±0.2°、23.6±0.2°、26.6±0.2°、13.8±0.2°或14.5±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；Or, the X-ray powder diffraction pattern of the crystal form B optionally further includes a position at 2θ of 22.2±0.2°, 22.4±0.2°, 22.9±0.2°, 23.6±0.2°, 26.6±0.2°, 13.8±0.2° Or one or more diffraction peaks at 14.5±0.2°; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5, 6, 7;
或，所述晶型C的X-射线粉末衍射图谱任选还包含位于2θ为14.7±0.2°、15.8±0.2°、18.0±0.2°、19.8±0.2°、23.6±0.2°、9.1±0.2°或16.7±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；Or, the X-ray powder diffraction pattern of the crystalline form C optionally further includes a position at 2θ of 14.7±0.2°, 15.8±0.2°, 18.0±0.2°, 19.8±0.2°, 23.6±0.2°, 9.1±0.2° Or one or more diffraction peaks at 16.7±0.2°; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5, 6, 7;
或，所述晶型D的X-射线粉末衍射图谱任选还包含位于2θ为17.2±0.2°、23.8±0.2°、25.5±0.2°、27.1±0.2°、15.3±0.2°、15.8±0.2°或21.4±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处。Or, the X-ray powder diffraction pattern of the crystal form D optionally further includes a position at 2θ of 17.2±0.2°, 23.8±0.2°, 25.5±0.2°, 27.1±0.2°, 15.3±0.2°, 15.8±0.2° Or one or more diffraction peaks at 21.4±0.2°; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5, 6, 7.
在本发明的优选方案中，所述晶型A的X-射线粉末衍射图谱任选包含位于2θ为7.8±0.2°、14.1±0.2°、15.3±0.2°、19.5±0.2°、12.0±0.2°、12.4±0.2°、13.2±0.2°、14.6±0.2°、21.2±0.2°、22.0±0.2°、22.6±0.2°、25.0±0.2°、28.3±0.2°、16.7±0.2°或24.5±0.2°处中的一处或多处衍射峰，In a preferred embodiment of the present invention, the X-ray powder diffraction pattern of the crystalline form A optionally includes 2θ at 7.8±0.2°, 14.1±0.2°, 15.3±0.2°, 19.5±0.2°, 12.0±0.2° , 12.4±0.2°, 13.2±0.2°, 14.6±0.2°, 21.2±0.2°, 22.0±0.2°, 22.6±0.2°, 25.0±0.2°, 28.3±0.2°, 16.7±0.2° or 24.5±0.2° One or more diffraction peaks in
优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 places;
或，所述晶型B的X-射线粉末衍射图谱任选包含位于2θ为4.2±0.2°、12.2±0.2°、12.6±0.2°、16.5±0.2°、16.8±0.2°、18.4±0.2°、18.9±0.2°、21.1±0.2°、22.2±0.2°、22.4±0.2°、22.9±0.2°、23.6±0.2°、26.6±0.2°、13.8±0.2°或14.5±0.2°处中的一处或多处衍射峰，Or, the X-ray powder diffraction pattern of the crystal form B optionally includes a position at 2θ of 4.2±0.2°, 12.2±0.2°, 12.6±0.2°, 16.5±0.2°, 16.8±0.2°, 18.4±0.2°, One of 18.9±0.2°, 21.1±0.2°, 22.2±0.2°, 22.4±0.2°, 22.9±0.2°, 23.6±0.2°, 26.6±0.2°, 13.8±0.2° or 14.5±0.2°, or multiple diffraction peaks,
优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；例如，所述晶型B的X-射线粉末衍射图谱在2θ为以下位置处有衍射峰：Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 positions; for example, the X-ray powder diffraction pattern of the crystal form B has diffraction peaks at the following positions at 2θ:
或，所述晶型C的X-射线粉末衍射图谱任选包含位于2θ为11.4±0.2°、12.5±0.2°、21.1±0.2°、23.0±0.2°、26.1±0.2°、26.6±0.2°、13.0±0.2°、14.0±0.2°、14.7±0.2°、15.8±0.2°、18.0±0.2°、19.8±0.2°、23.6±0.2°、9.1±0.2°或16.7±0.2°处中的一处或多处衍射峰，Or, the X-ray powder diffraction pattern of the crystal form C optionally includes a position at 2θ of 11.4±0.2°, 12.5±0.2°, 21.1±0.2°, 23.0±0.2°, 26.1±0.2°, 26.6±0.2°, One of 13.0±0.2°, 14.0±0.2°, 14.7±0.2°, 15.8±0.2°, 18.0±0.2°, 19.8±0.2°, 23.6±0.2°, 9.1±0.2° or 16.7±0.2° or multiple diffraction peaks,
优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 places;
或，所述晶型D的X-射线粉末衍射图谱任选包含位于2θ为9.5±0.2°、10.4±0.2°、14.5±0.2°、19.9±0.2°、20.8±0.2°、24.8±0.2°、11.7±0.2°、13.3±0.2°、17.2±0.2°、23.8±0.2°、25.5±0.2°、27.1±0.2°、15.3±0.2°、15.8±0.2°或21.4±0.2°处中的一处或多处衍射峰，Or, the X-ray powder diffraction pattern of the crystal form D optionally includes a position at 2θ of 9.5±0.2°, 10.4±0.2°, 14.5±0.2°, 19.9±0.2°, 20.8±0.2°, 24.8±0.2°, One of 11.7±0.2°, 13.3±0.2°, 17.2±0.2°, 23.8±0.2°, 25.5±0.2°, 27.1±0.2°, 15.3±0.2°, 15.8±0.2° or 21.4±0.2° or multiple diffraction peaks,
优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰。Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 positions.
在本发明进一步优选的实施方式中，所述晶型为N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型B；In a further preferred embodiment of the present invention, the crystal form is N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methanol Base)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5 , the crystal form B of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
所述晶型B的X-射线粉末衍射图谱在4.0±0.2°处具有衍射峰；或者在12.0±0.2°处具有衍射峰；或者在12.4±0.2°处具有衍射峰；或者在16.4±0.2°处具有衍射峰；或者在16.6±0.2°处具有衍射峰；或者在18.2±0.2°处具有衍射峰；或者在18.7±0.2°处具有衍射峰；或者在20.9±0.2°处具有衍射峰；或者在22.0±0.2°处具有衍射峰；或者在22.3±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form B has a diffraction peak at 4.0±0.2°; or has a diffraction peak at 12.0±0.2°; or has a diffraction peak at 12.4±0.2°; or has a diffraction peak at 16.4±0.2° or have a diffraction peak at 16.6±0.2°; or have a diffraction peak at 18.2±0.2°; or have a diffraction peak at 18.7±0.2°; or have a diffraction peak at 20.9±0.2°; or Have a diffraction peak at 22.0±0.2°; or have a diffraction peak at 22.3±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3-8 of the above-mentioned diffraction peaks , or 5-8, or 6-8, more preferably including any 6, 7 or 8 of them;
优选地，所述晶型B的X-射线粉末衍射图谱至少包含位于2θ为4.0±0.2°、12.0±0.2°、12.4±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含16.4±0.2°、16.6±0.2°、18.2±0.2°、18.7±0.2°、20.9±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；Preferably, the X-ray powder diffraction pattern of the crystal form B includes at least one or more diffraction peaks at 2θ of 4.0±0.2°, 12.0±0.2°, and 12.4±0.2°, preferably two of them, More preferably, it contains three; optionally, it can further include at least one of 16.4±0.2°, 16.6±0.2°, 18.2±0.2°, 18.7±0.2°, 20.9±0.2°, preferably 2 or 3 of them , 4 or 5;
更优选地，所述晶型B的X-射线粉末衍射图谱任选还包含位于2θ为22.0±0.2°、22.3±0.2°、22.7±0.2°、23.5±0.2°、26.5±0.2°、13.6±0.2°或14.3±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；More preferably, the X-ray powder diffraction pattern of the crystal form B optionally further comprises One or more diffraction peaks at 0.2° or 14.3±0.2°; preferably at least include any 2-3, or 4-5, or 6-7 of them; more preferably, include any 2 or 3 of them place, 4 places, 5 places, 6 places, 7 places;
进一步优选地，所述晶型B的X-射线粉末衍射图谱任选包含位于2θ为4.0±0.2°、12.0±0.2°、12.4±0.2°、16.4±0.2°、16.6±0.2°、18.2±0.2°、18.7±0.2°、20.9±0.2°、22.0±0.2°、22.3±0.2°、22.7±0.2°、23.5±0.2°、26.5±0.2°、13.6±0.2°或14.3±0.2°处中的一处或多处衍射峰，优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；Further preferably, the X-ray powder diffraction pattern of the crystal form B optionally includes °, 18.7±0.2°, 20.9±0.2°, 22.0±0.2°, 22.3±0.2°, 22.7±0.2°, 23.5±0.2°, 26.5±0.2°, 13.6±0.2° or 14.3±0.2° One or more diffraction peaks, preferably, including optional 4, 5, 6, 8 or 10 diffraction peaks;
更进一步优选地，所述晶型B的X-射线粉末衍射图谱如图10所示。Even more preferably, the X-ray powder diffraction pattern of the crystal form B is shown in FIG. 10 .
在本发明进一步优选的实施方式中，实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型A，使用Cu-Kα辐射，以2θ角和晶面间距d值表示的X-射线特征衍射峰如表1所示。In a further preferred embodiment of the present invention, Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form A of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2θ angles and interplanar spacing d values using Cu-Kα radiation The diffraction peaks are shown in Table 1.
表1实施例4化合物自由碱晶型A的XRPD射线衍射数据Table 1 Example 4 The XRPD ray diffraction data of compound free base crystal form A
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(I％)Proportion (I%) 面积area 比例(I％)Proportion (I%)
11 7.777.77 11.36911.369 402402 100100 53565356 50.450.4
22 9.3769.376 9.42429.4242 5252 12.912.9 383383 3.63.6
33 12.00812.008 7.36457.3645 167167 41.541.5 17801780 16.716.7
44 12.43612.436 7.11187.1118 139139 34.634.6 14861486 1414
55 13.22713.227 6.68836.6883 9696 23.923.9 908908 8.58.5
66 14.07214.072 6.28856.2885 260260 64.764.7 27872787 26.226.2
77 14.60214.602 6.06156.0615 142142 35.335.3 15571557 14.614.6
88 15.12915.129 5.85125.8512 204204 50.750.7 42244224 39.739.7
99 15.31415.314 5.78125.7812 294294 73.173.1 61216121 57.557.5
1010 16.00116.001 5.53425.5342 7171 17.717.7 729729 6.96.9
1111 16.67316.673 5.31285.3128 129129 32.132.1 31743174 29.829.8
1212 17.17917.179 5.15735.1573 101101 25.125.1 22492249 21.121.1
1313 18.0918.09 4.89974.8997 109109 27.127.1 12421242 11.711.7
1414 18.85718.857 4.70214.7021 5959 14.714.7 453453 4.34.3
1515 19.46919.469 4.55564.5556 364364 90.590.5 1063710637 100100
1616 19.91519.915 4.45464.4546 221221 5555 72207220 67.967.9
1717 20.54820.548 4.31894.3189 6363 15.715.7 441441 4.14.1
1818 21.19521.195 4.18834.1883 162162 40.340.3 18861886 17.717.7
1919 22.02322.023 4.03274.0327 210210 52.252.2 33133313 31.131.1
2020 22.5922.59 3.93293.9329 197197 4949 41524152 3939
21twenty one 23.46323.463 3.78853.7885 6262 15.415.4 13291329 12.512.5
22twenty two 23.92923.929 3.71573.7157 5656 13.913.9 10121012 9.59.5
23twenty three 24.45524.455 3.63693.6369 114114 28.428.4 14491449 13.613.6
24twenty four 25.02525.025 3.55533.5553 196196 48.848.8 27032703 25.425.4
2525 25.81325.813 3.44863.4486 6868 16.916.9 23992399 22.622.6
2626 26.03726.037 3.41953.4195 9090 22.422.4 30273027 28.528.5
2727 26.34226.342 3.38063.3806 117117 29.129.1 31863186 3030
2828 28.32428.324 3.14833.1483 151151 37.637.6 24652465 23.223.2
2929 29.36129.361 3.03943.0394 4545 11.211.2 533533 55
3030 30.78430.784 2.90212.9021 7878 19.419.4 936936 8.88.8
3131 31.53131.531 2.8352.835 5151 12.712.7 12521252 11.811.8
3232 31.71231.712 2.81932.8193 4949 12.212.2 967967 9.19.1
3333 33.17133.171 2.69862.6986 4444 10.910.9 956956 99
本发明实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型A，其X-射线粉末衍射图谱基本如图1所示；其DSC图谱基本如图2所示。Compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine in Example 4 of the present invention -2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H - Crystal form A of imidazo[4,5-c]pyridine-2-carboxamide, its X-ray powder diffraction pattern is basically shown in Figure 1; its DSC pattern is basically shown in Figure 2.
在本发明进一步优选的实施方式中，实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型B，使用Cu-Kα辐射，以2θ角和晶面间距d值表示的X-射线特征衍射峰如表2所示。In a further preferred embodiment of the present invention, Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form B of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2θ angles and interplanar spacing d values using Cu-Kα radiation The diffraction peaks are shown in Table 2.
表2实施例4化合物自由碱晶型B的XRPD射线衍射数据The XRPD ray diffraction data of table 2 embodiment 4 compound free base crystal form B
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(I％)Proportion (I%) 面积area 比例(I％)Proportion (I%)
11 4.1834.183 21.107221.1072 661661 49.749.7 70327032 63.763.7
22 8.728.72 10.132410.1324 135135 10.210.2 929929 8.48.4
33 9.7519.751 9.0639.063 8989 6.76.7 588588 5.35.3
44 11.80611.806 7.497.49 251251 18.918.9 15901590 14.414.4
55 12.15112.151 7.27797.2779 547547 41.141.1 46974697 42.642.6
66 12.55512.555 7.04457.0445 13301330 100100 1081410814 9898
77 13.7913.79 6.41646.4164 214214 16.116.1 19091909 17.317.3
88 14.48114.481 6.11166.1116 296296 22.322.3 19191919 17.417.4
99 14.97214.972 5.91255.9125 395395 29.729.7 33973397 30.830.8
1010 15.27615.276 5.79535.7953 8989 6.76.7 886886 88
1111 16.03616.036 5.52245.5224 245245 18.418.4 25622562 23.223.2
1212 16.50816.508 5.36555.3655 716716 53.853.8 62746274 56.856.8
1313 16.78916.789 5.27625.2762 836836 62.962.9 61896189 56.156.1
1414 17.50517.505 5.0625.062 436436 32.832.8 42114211 38.238.2
1515 17.98317.983 4.92854.9285 263263 19.819.8 20572057 18.618.6
1616 18.39218.392 4.81994.8199 619619 46.546.5 41284128 37.437.4
1717 18.89718.897 4.69224.6922 587587 44.144.1 51385138 46.646.6
1818 19.46519.465 4.55654.5565 131131 9.89.8 668668 6.16.1
1919 20.45920.459 4.33734.3373 445445 33.533.5 49434943 44.844.8
2020 21.06721.067 4.21364.2136 551551 41.441.4 60216021 54.654.6
21twenty one 21.63821.638 4.10364.1036 133133 1010 725725 6.66.6
22twenty two 22.18222.182 4.00434.0043 821821 61.761.7 94589458 85.785.7
23twenty three 22.42622.426 3.96113.9611 825825 6262 81928192 74.274.2
24twenty four 22.89722.897 3.88083.8808 619619 46.546.5 47714771 43.243.2
2525 23.64723.647 3.75943.7594 13271327 99.899.8 1103711037 100100
2626 24.59624.596 3.61653.6165 286286 21.521.5 39963996 36.236.2
2727 24.824.8 3.58723.5872 327327 24.624.6 42224222 38.338.3
2828 25.2125.21 3.52973.5297 204204 15.315.3 82418241 74.774.7
2929 25.4525.45 3.49693.4969 437437 32.932.9 61376137 55.655.6
3030 26.64726.647 3.34263.3426 644644 48.448.4 77317731 7070
3131 27.41427.414 3.25073.2507 5757 4.34.3 744744 6.76.7
3232 27.86227.862 3.19953.1995 6464 4.84.8 765765 6.96.9
3333 28.54928.549 3.1243.124 9494 7.17.1 989989 99
3434 28.97728.977 3.07883.0788 142142 10.710.7 14721472 13.313.3
3535 29.4629.46 3.02943.0294 124124 9.39.3 17261726 15.615.6
3636 29.98929.989 2.97722.9772 102102 7.77.7 16891689 15.315.3
3737 30.60330.603 2.91882.9188 9292 6.96.9 796796 7.27.2
3838 31.28831.288 2.85652.8565 6969 5.25.2 667667 66
3939 32.24132.241 2.77422.7742 8585 6.46.4 10351035 9.49.4
4040 32.98932.989 2.7132.713 135135 10.210.2 16561656 1515
4141 34.40834.408 2.60432.6043 6565 4.94.9 490490 4.44.4
4242 35.05735.057 2.55762.5576 107107 88 14341434 1313
4343 35.49935.499 2.52672.5267 8181 6.16.1 866866 7.87.8
4444 36.31636.316 2.47172.4717 4747 3.53.5 11391139 10.310.3
4545 36.58836.588 2.45392.4539 6565 4.94.9 11341134 10.310.3
4646 38.18238.182 2.35512.3551 5858 4.44.4 10421042 9.49.4
4747 38.47938.479 2.33762.3376 5050 3.83.8 10461046 9.59.5
本发明实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型B，其X-射线粉末衍射图谱基本如图3所示；其DSC图谱基本如图4所示。Compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine in Example 4 of the present invention -2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H - The crystal form B of imidazo[4,5-c]pyridine-2-carboxamide, its X-ray powder diffraction pattern is basically shown in Figure 3; its DSC pattern is basically shown in Figure 4.
在本发明进一步优选的实施方式中，实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型C，使用Cu-Kα辐射，以2θ角和晶面间距d值表示的X-射线特征衍射峰如表3所示。In a further preferred embodiment of the present invention, Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form C of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2θ angles and interplanar spacing d values using Cu-Kα radiation The diffraction peaks are shown in Table 3.
表3实施例4化合物自由碱晶型C的XRPD射线衍射数据Table 3 Example 4 The XRPD ray diffraction data of the free base crystal form C of the compound
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(I％)Proportion (I%) 面积area 比例(I％)Proportion (I%)
11 7.5687.568 11.671711.6717 8282 17.517.5 940940 8.48.4
22 7.9327.932 11.137211.1372 8484 17.917.9 11491149 10.310.3
33 9.0659.065 9.74729.7472 7373 15.615.6 248248 2.22.2
44 11.40111.401 7.75517.7551 391391 83.583.5 49924992 44.844.8
55 12.49712.497 7.07697.0769 358358 76.576.5 30663066 27.527.5
66 13.00313.003 6.8036.803 170170 36.336.3 46464646 41.741.7
77 13.30413.304 6.64946.6494 6666 14.114.1 19851985 17.817.8
88 13.95313.953 6.34166.3416 117117 2525 837837 7.57.5
99 14.33814.338 6.17226.1722 110110 23.523.5 27352735 24.624.6
1010 14.66414.664 6.03576.0357 234234 5050 63196319 56.756.7
1111 14.96814.968 5.9145.914 114114 24.424.4 35673567 3232
1212 15.81715.817 5.59855.5985 151151 32.332.3 25912591 23.323.3
1313 16.30116.301 5.43335.4333 6161 1313 10621062 9.59.5
1414 16.72916.729 5.29525.2952 7676 16.216.2 715715 6.46.4
1515 18.03118.031 4.91564.9156 150150 32.132.1 40054005 3636
1616 18.49918.499 4.79234.7923 7070 1515 19251925 17.317.3
1717 19.81519.815 4.47694.4769 217217 46.446.4 65566556 58.958.9
1818 20.25720.257 4.38024.3802 100100 21.421.4 46234623 41.541.5
1919 20.68520.685 4.29044.2904 8080 17.117.1 14141414 12.712.7
2020 21.13121.131 4.2014.201 231231 49.449.4 27872787 2525
21twenty one 22.95622.956 3.87093.8709 468468 100100 99649964 89.589.5
22twenty two 23.63823.638 3.76073.7607 147147 31.431.4 14901490 13.413.4
23twenty three 24.53724.537 3.6253.625 7373 15.615.6 445445 44
24twenty four 26.11726.117 3.40923.4092 347347 74.174.1 1113911139 100100
2525 26.64426.644 3.34283.3428 279279 59.659.6 69906990 62.862.8
2626 27.51727.517 3.23883.2388 9090 19.219.2 19721972 17.717.7
2727 27.90727.907 3.19443.1944 8484 17.917.9 21382138 19.219.2
2828 29.32329.323 3.04333.0433 8686 18.418.4 23412341 21twenty one
2929 29.60429.604 3.01513.0151 9494 20.120.1 34533453 3131
3030 31.28631.286 2.85672.8567 101101 21.621.6 17351735 15.615.6
3131 32.28232.282 2.77082.7708 5858 12.412.4 656656 5.95.9
3232 33.0133.01 2.71132.7113 5454 11.511.5 11991199 10.810.8
3333 33.37933.379 2.68222.6822 6363 13.513.5 15691569 14.114.1
3434 35.44835.448 2.53022.5302 5656 1212 965965 8.78.7
本发明实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型C，其X-射线粉末衍射图谱基本如图5所示；其DSC图谱基本如图6所示。Compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine in Example 4 of the present invention -2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H - Form C of imidazo[4,5-c]pyridine-2-carboxamide, its X-ray powder diffraction pattern is basically shown in Figure 5; its DSC pattern is basically shown in Figure 6.
在本发明进一步优选的实施方式中，实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型D，使用Cu-Kα辐射，以2θ角和晶面间距d值表示的X-射线特征衍射峰如表4所示。In a further preferred embodiment of the present invention, Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5, Form D of 6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide, X-ray characterization in terms of 2θ angles and interplanar spacing d values using Cu-Kα radiation The diffraction peaks are shown in Table 4.
表4实施例4化合物自由碱晶型D的XRPD射线衍射数据Table 4 Example 4 The XRPD ray diffraction data of the free base crystal form D of the compound
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(I％)Proportion (I%) 面积area 比例(I％)Proportion (I%)
11 6.0196.019 14.671514.6715 6666 9.79.7 130130 1.71.7
22 8.7688.768 10.077110.0771 6161 99 458458 5.85.8
33 9.4549.454 9.34729.3472 681681 100100 78477847 100100
44 10.42710.427 8.47718.4771 305305 44.844.8 33403340 42.642.6
55 11.66411.664 7.58097.5809 167167 24.524.5 22272227 28.428.4
66 13.26513.265 6.66886.6688 134134 19.719.7 22542254 28.728.7
77 13.75413.754 6.43326.4332 7272 10.610.6 11411141 14.514.5
88 14.45814.458 6.12136.1213 331331 48.648.6 45094509 57.557.5
99 15.31715.317 5.77985.7798 101101 14.814.8 13301330 16.916.9
1010 15.84215.842 5.58965.5896 100100 14.714.7 12421242 15.815.8
1111 17.21617.216 5.14645.1464 128128 18.818.8 17531753 22.322.3
1212 18.37118.371 4.82554.8255 5858 8.58.5 961961 12.212.2
1313 19.06119.061 4.65224.6522 5050 7.37.3 655655 8.38.3
1414 19.93519.935 4.45014.4501 296296 43.543.5 44674467 56.956.9
1515 20.80720.807 4.26564.2656 225225 3333 26152615 33.333.3
1616 21.43621.436 4.14194.1419 108108 15.915.9 36693669 46.846.8
1717 22.00222.002 4.03664.0366 6262 9.19.1 13521352 17.217.2
1818 22.77222.772 3.90183.9018 6363 9.39.3 458458 5.85.8
1919 23.36523.365 3.80413.8041 9292 13.513.5 14061406 17.917.9
2020 23.78523.785 3.73783.7378 189189 27.827.8 23662366 30.230.2
21twenty one 24.80124.801 3.58693.5869 331331 48.648.6 54865486 69.969.9
22twenty two 25.48825.488 3.49183.4918 197197 28.928.9 25552555 32.632.6
23twenty three 26.16326.163 3.40333.4033 107107 15.715.7 17401740 22.222.2
24twenty four 27.11627.116 3.28583.2858 171171 25.125.1 27792779 35.435.4
2525 27.5427.54 3.23613.2361 7878 11.511.5 11491149 14.614.6
2626 28.81528.815 3.09583.0958 105105 15.415.4 16841684 21.521.5
2727 29.71129.711 3.00443.0044 7272 10.610.6 14761476 18.818.8
2828 31.8631.86 2.80652.8065 5151 7.57.5 986986 12.612.6
本发明实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型D，其X-射线粉末衍射图谱基本如图7所示；其DSC图谱基本如图8所示。Compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine in Example 4 of the present invention -2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H - The crystal form D of imidazo[4,5-c]pyridine-2-carboxamide, its X-ray powder diffraction pattern is basically shown in Figure 7; its DSC pattern is basically shown in Figure 8.
在本发明进一步优选的方案中，实施例4化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型A、晶型B、晶型C和晶型D的X-射线粉末衍射图谱中相对峰强度为前十强的衍射峰位置与图1、图3、图5和图7对应位置衍射峰的2θ误差为±0.2°～±0.5°，优选±0.2°～±0.3°， 最优选±0.2°；In a further preferred embodiment of the present invention, Example 4 compound N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl) -6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6 , Relative peak intensities in the X-ray powder diffraction patterns of 7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide in Form A, Form B, Form C and Form D The 2θ error between the positions of the top ten diffraction peaks and the corresponding positions in Figure 1, Figure 3, Figure 5 and Figure 7 is ±0.2°～±0.5°, preferably ±0.2°～±0.3°, most preferably ±0.2° ;
本发明所述的误差是指由于实验过程中制样技术水平差异、操作的规范性、样品制样的厚度、使用不同的样品架等实验误差因素导致的不同批次样品的X-射线粉末衍射图谱中的衍射峰位置产生一定程度的整体漂移，对于本领域技术人员来说由于实验误差导致的衍射峰位置整体漂移都是可以预料得到的，下表5、6、7与8是四个不同批次晶型B的X-射线特征衍射峰数据表，其X-射线粉末衍射图谱基本如图9、10、11和12所示：The error described in the present invention refers to the X-ray powder diffraction of different batches of samples caused by experimental error factors such as the difference in the level of sample preparation technology, the standardization of operation, the thickness of sample sample preparation, and the use of different sample racks during the experiment. The position of the diffraction peak in the spectrum has a certain degree of overall drift. For those skilled in the art, the overall drift of the diffraction peak position due to experimental errors is predictable. The following tables 5, 6, 7 and 8 are four different The X-ray characteristic diffraction peak data table of the batch crystal form B, and its X-ray powder diffraction patterns are basically shown in Figures 9, 10, 11 and 12:
表5自由碱晶型B批次一的XRPD射线衍射数据Table 5 XRPD ray diffraction data of free alkali crystal form B batch 1
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(％)Proportion(%)
11 3.7473.747 23.5628523.56285 13521352 78.778.7
22 3.9013.901 22.6319922.63199 17191719 100100
33 7.6297.629 11.5783011.57830 519519 30.230.2
44 8.4378.437 10.4715310.47153 386386 22.522.5
55 9.4759.475 9.326499.32649 317317 18.518.5
66 11.56011.560 7.648637.64863 478478 27.827.8
77 11.88211.882 7.441947.44194 736736 42.842.8
88 12.28912.289 7.196817.19681 12371237 72.072.0
99 12.92412.924 6.844646.84464 275275 16.016.0
1010 13.53713.537 6.535896.53589 392392 22.822.8
1111 13.85313.853 6.387366.38736 319319 18.618.6
1212 14.22114.221 6.223166.22316 408408 23.723.7
1313 14.73214.732 6.011836.01183 511511 29.729.7
1414 15.05115.051 5.881495.88149 450450 26.226.2
1515 15.76515.765 5.616795.61679 452452 26.326.3
1616 16.24116.241 5.453195.45319 10171017 59.259.2
1717 16.51216.512 5.364505.36450 780780 45.445.4
1818 17.23317.233 5.141545.14154 485485 28.228.2
1919 17.71517.715 5.002615.00261 413413 24.024.0
2020 18.13118.131 4.888784.88878 651651 37.937.9
21twenty one 18.62818.628 4.759564.75956 712712 41.441.4
22twenty two 18.85818.858 4.701894.70189 318318 18.518.5
23twenty three 19.22319.223 4.613534.61353 361361 21.021.0
24twenty four 19.46219.462 4.557384.55738 337337 19.619.6
2525 20.19720.197 4.393124.39312 558558 32.532.5
2626 20.79920.799 4.267344.26734 683683 39.839.8
2727 21.37321.373 4.154054.15405 316316 14.814.8
2828 21.91521.915 4.052514.05251 12581258 73.273.2
2929 22.15922.159 4.008334.00833 956956 55.655.6
3030 22.62922.629 3.926293.92629 724724 42.142.1
3131 23.40023.400 3.798493.79849 14061406 81.881.8
3232 24.35124.351 3.652333.65233 485485 28.228.2
3333 24.52924.529 3.626253.62625 588588 34.234.2
3434 25.17825.178 3.534143.53414 647647 37.737.7
3535 26.39626.396 3.373793.37379 811811 47.247.2
3636 27.16027.160 3.280593.28059 253253 14.714.7
3737 28.23728.237 3.157903.15790 311311 18.118.1
3838 28.72528.725 3.105353.10535 326326 19.019.0
3939 29.20529.205 3.055373.05537 251251 14.614.6
4040 29.73629.736 3.002053.00205 262262 15.215.2
4141 30.35330.353 2.942372.94237 232232 13.513.5
4242 31.01831.018 2.880582.88058 240240 14.014.0
4343 31.56431.564 2.832212.83221 309309 18.018.0
4444 31.99931.999 2.794742.79474 239239 13.913.9
4545 32.71732.717 2.734982.73498 260260 15.115.1
4646 34.07734.077 2.628902.62890 179179 10.410.4
4747 34.80734.807 2.575392.57539 213213 12.412.4
4343 35.22335.223 2.545952.54595 183183 10.610.6
4545 36.34536.345 2.469842.46984 207207 12.112.1
4646 45.30945.309 1.999881.99988 209209 12.212.2
表6自由碱晶型B批次二的XRPD射线衍射数据Table 6 XRPD ray diffraction data of free base crystal form B batch 2
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(％)Proportion(%)
11 3.6153.615 24.4202524.42025 299299 12.412.4
22 4.0544.054 21.7793521.77935 15121512 62.662.6
33 8.1228.122 10876751087675 259259 10.710.7
44 8.5798.579 10.2991510.29915 452452 18.718.7
55 9.5969.596 9.209789.20978 358358 14.814.8
66 11.67811.678 7.571657.57165 584584 24.224.2
77 12.01212.012 7.362047.36204 10371037 42.942.9
88 12.41212.412 7.125417.12541 24172417 100100
99 13.63413.634 6.489636.48963 576576 23.823.8
1010 14.11114.111 6.271046.27104 277277 11.511.5
1111 14.33514.335 6.173616.17361 585585 24.224.2
1212 14.83914.839 5.965245.96524 728728 30.130.1
1313 15.16115.161 5.839035.83903 338338 14.014.0
1414 15.87815.878 5.577115.57711 503503 20.820.8
1515 16.02816.028 5.525315.52531 382382 15.815.8
1616 16.36816.368 5.411225.41122 10821082 44.844.8
1717 16.63416.634 5.325425.32542 12561256 52.052.0
1818 17.25417.254 5.135295.13529 589589 24.424.4
1919 17.37017.370 5.101255.10125 733733 30.330.3
2020 17.82817.828 4.971324.97132 594594 24.624.6
21twenty one 18.23718.237 4.860584.86058 910910 37.637.6
22twenty two 18.73218.732 4.733234.73323 986986 40.840.8
23twenty three 18.96018.960 4.676794.67679 366366 15.115.1
24twenty four 19.30119.301 4.595104.59510 330330 13.713.7
2525 20.30320.303 4.370514.37051 841841 34.834.8
2626 20.45920.459 4.337574.33757 419419 17.317.3
2727 20.75820.758 4.275764.27576 609609 25.225.2
2828 20.90820.908 4.245304.24530 848848 35.135.1
2929 21.49221.492 4.131334.13133 318318 13.213.2
3030 22.02022.020 4.033474.03347 12541254 51.951.9
3131 22.26622.266 3.989463.98946 11361136 47.047.0
3232 22.74222.742 3.906913.90691 964964 39.939.9
3333 23.50423.504 3.781953.78195 16761676 69.369.3
3434 24.22524.225 3.670963.67096 310310 12.812.8
3535 24.43824.438 3.639453.63945 578578 23.923.9
3636 24.64524.645 3.609353.60935 591591 24.524.5
3737 24.91324.913 3.571243.57124 475475 19.719.7
3838 25.28925.289 3.518883.51888 722722 29.929.9
3939 25.98525.985 3.426243.42624 272272 11.311.3
4040 26.49826.498 3.361003.36100 924924 38.238.2
4141 27.12927.129 3.284363.28436 227227 9.49.4
4242 27.29027.290 3.265353.26535 241241 10.010.0
4343 27.69927.699 3.217983.21798 263263 10.910.9
4444 28.82528.825 3.094833.09483 370370 15.315.3
4545 29.30129.301 3.045653.04565 284284 11.811.8
4646 29.82029.820 2.993742.99374 278278 11.511.5
4747 30.45530.455 2.932812.93281 260260 10.810.8
4343 31.09431.094 2.873922.87392 221221 9.19.1
4545 32.05632.056 2.789832.78983 253253 10.510.5
4646 32.80332.803 2.728042.72804 303303 12.512.5
4141 34.23334.233 2.617282.61728 205205 8.58.5
4242 34.83834.838 2.573212.57321 233233 9.69.6
4343 35.34035.340 2.537752.53775 215215 8.98.9
4444 36.11636.116 2.484992.48499 193193 8.08.0
4545 36.41036.410 2.465642.46564 198198 8.28.2
4646 37.21137.211 2.414342.41434 223223 9.29.2
4747 47.08847.088 1.928391.92839 156156 6.56.5
4848 47.96847.968 1.895031.89503 178178 7.47.4
4949 50.01050.010 1.822361.82236 147147 6.16.1
5050 52.26952.269 1.748791.74879 129129 5.35.3
表7自由碱晶型B批次三的XRPD射线衍射数据Table 7 XRPD ray diffraction data of free base crystal form B batch 3
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(％)Proportion(%)
11 3.6093.609 24.4655924.46559 278278 8.58.5
22 3.9733.973 22.2209422.22094 32673267 100100
33 8.0578.057 10.9642110.96421 359359 11.011.0
44 8.4878.487 10.4101910.41019 643643 19.719.7
55 9.5039.503 9.299639.29963 434434 13.313.3
66 11.59011.590 7.628967.62896 10081008 30.930.9
77 11.91811.918 7.419967.41996 12341234 37.837.8
88 12.31412.314 7.182247.18224 19131913 58.658.6
99 13.53213.532 6.538346.53834 474474 14.514.5
1010 14.23514.235 6.216766.21676 495495 15.215.2
1111 14.74114.741 6.004536.00453 692692 21.221.2
1212 15.07215.072 5.873305.87330 321321 9.89.8
1313 15.77915.779 5.611915.61191 451451 13.813.8
1414 15.93815.938 5.556285.55628 324324 9.99.9
1515 16.26716.267 5.444585.44458 19621962 60.160.1
1616 16.53516.535 5.356845.35684 10651065 32.632.6
1717 17.16317.163 5.162315.16231 619619 18.918.9
1818 17.26717.267 5.131575.13157 696696 21.321.3
1919 17.73517.735 4.997154.99715 707707 21.621.6
2020 18.15818.158 4.881724.88172 10591059 32.432.4
21twenty one 18.64218.642 4.755844.75584 11021102 33.733.7
22twenty two 18.85818.858 4.702084.70208 320320 9.89.8
23twenty three 19.21019.210 4.616544.61654 384384 11.811.8
24twenty four 20.20720.207 4.390984.39098 747747 22.922.9
2525 20.39420.394 4.351094.35109 705705 21.621.6
2626 20.65620.656 4.296524.29652 634634 19.419.4
2727 20.82820.828 4.261454.26145 16961696 51.951.9
2828 21.39421.394 4.150034.15003 345345 10.610.6
2929 21.93321.933 4.049174.04917 20372037 62.462.4
3030 22.17922.179 4.004874.00487 13121312 40.240.2
3131 22.65022.650 3.922673.92267 985985 30.130.1
3232 23.42523.425 3.794533.79453 31553155 96.696.6
3333 24.33524.335 3.654743.65474 581581 17.817.8
3434 24.55924.559 3.621883.62188 887887 27.227.2
3535 24.81124.811 3.585583.58558 521521 15.915.9
3636 25.02825.028 3.555113.55511 571571 17.517.5
3737 25.20625.206 3.530393.53039 832832 25.525.5
3838 25.83425.834 3.445973.44597 318318 9.79.7
3939 25.90525.905 3.436683.43668 307307 9.49.4
4040 26.14226.142 3.405963.40596 495495 15.215.2
4141 26.43226.432 3.369363.36936 18921892 57.957.9
4242 27.17827.178 3.278453.27845 224224 6.96.9
4343 27.61527.615 3.227653.22765 225225 6.96.9
4444 28.21728.217 3.160143.16014 241241 7.47.4
4545 28.74828.748 3.102933.10293 451451 13.813.8
4646 29.25029.250 3.050843.05084 279279 8.58.5
4747 29.74229.742 3.001463.00146 343343 10.510.5
4343 30.38530.385 2.939362.93936 330330 10.110.1
4545 31.02931.029 2.879862.87986 242242 7.47.4
4646 31.99131.991 2.795342.79534 270270 8.38.3
4040 32.73232.732 2.733742.73374 303303 9.39.3
4141 34.14134.141 2.624132.62413 207207 6.36.3
4242 34.81934.819 2.574512.57451 309309 9.59.5
4343 35.23935.239 2.544792.54479 212212 6.56.5
4444 36.08536.085 2.487062.48706 230230 7.07.0
4545 37.00137.001 2.427562.42756 199199 6.16.1
4646 43.09943.099 2.097162.09716 210210 6.46.4
表8自由碱晶型B批次四的XRPD射线衍射数据Table 8 XRPD ray diffraction data of free base crystal form B batch 4
序号serial number 2θ(±0.2°)2θ(±0.2°) d值d value 峰高Peak height 比例(％)Proportion(%)
11 4.0394.039 21.8570721.85707 47914791 100.0100.0
22 8.1198.119 10.8810510.88105 394394 8.28.2
33 8.5688.568 10.3123510.31235 559559 11.711.7
44 9.3899.389 9.411649.41164 250250 5.25.2
55 9.5869.586 9.218829.21882 389389 8.18.1
66 11.66811.668 7.577927.57792 10271027 21.421.4
77 12.00012.000 7.369547.36954 13791379 28.828.8
88 12.38912.389 7.138837.13883 14231423 29.729.7
99 13.61813.618 6.497356.49735 350350 7.37.3
1010 14.31914.319 6.180626.18062 386386 8.18.1
1111 14.80814.808 5.977635.97763 562562 11.711.7
1212 15.15615.156 5.841075.84107 323323 6.76.7
1313 15.85515.855 50585025058502 414414 8.68.6
1414 16.34516.345 5.418655.41865 29252925 61.161.1
1515 16.61216.612 5.332285.33228 945945 19.719.7
1616 17.25117.251 5.136295.13629 526526 11.011.0
1717 17.34817.348 5.107825.10782 576576 12.012.0
1818 17.61117.611 5.031945.03194 287287 6.06.0
1919 17.81917.819 4.973684.97368 699699 14.614.6
2020 18.25318.253 4.856344.85634 10901090 22.822.8
21twenty one 18.71918.719 4.736534.73653 985985 20.620.6
22twenty two 18.92518.925 4.685544.68554 299299 6.26.2
23twenty three 19.28619.286 4.598494.59849 367367 7.77.7
24twenty four 20.29320.293 4.372594.37259 668668 13.913.9
2525 20.47920.479 4.333374.33337 10291029 21.521.5
2626 20.72120.721 4.283294.28329 501501 10.510.5
2727 20.91320.913 4.244294.24429 17621762 36.836.8
2828 21.47321.473 4.134864.13486 351351 7.37.3
2929 22.02122.021 4.033284.03328 31013101 64.764.7
3030 22.25922.259 3.990593.99059 13471347 28.128.1
3131 22.72822.728 3.909303.90930 858858 17.917.9
3232 23.50923.509 3.781243.78124 35483548 74.174.1
3333 24.41024.410 3.643693.64369 601601 12.512.5
3434 24.63924.639 3.610303.61030 13101310 27.327.3
3535 24.88624.886 3.575023.57502 548548 11.411.4
3636 25.10325.103 3.544633.54463 604604 12.612.6
3737 25.28725.287 3.519153.51915 875875 18.318.3
3838 25.62425.624 3.473703.47370 263263 5.55.5
3939 25.91825.918 3.434913.43491 349349 7.37.3
4040 26.24326.243 3.393093.39309 452452 9.49.4
4141 26.52026.520 3.358293.35829 21322132 44.544.5
4242 27.24727.247 3.270383.27038 237237 4.94.9
4343 27.69727.697 3.218253.21825 193193 4.04.0
4444 28.31928.319 3.148973.14897 288288 6.06.0
4545 28.38728.387 3.141523.14152 300300 6.36.3
4646 28.84228.842 3.093053.09305 508508 10.610.6
4747 29.83729.837 2.992072.99207 476476 9.99.9
4848 30.46130.461 2.932232.93223 340340 7.17.1
4949 31.11831.118 2.871762.87176 300300 6.36.3
5050 32.08632.086 2.787332.78733 316316 6.66.6
5151 32.80732.807 2.727672.72767 320320 6.76.7
5252 33.07733.077 2.706062.70606 191191 4.04.0
5353 33.50933.509 2.672152.67215 167167 3.53.5
5454 34.01834.018 2.633332.63333 205205 4.34.3
5555 34.23434.234 2.617212.61721 237237 4.94.9
5656 34.91134.911 2.568002.56800 311311 6.56.5
5757 35.78035.780 2.507542.50754 164164 3.43.4
5858 36.15136.151 2.482712.48271 199199 4.24.2
5959 36.43436.434 2.464052.46405 278278 5.85.8
6060 37.08237.082 2.422472.42247 233233 4.94.9
6161 38.66838.668 2.326692.32669 168168 3.53.5
6262 40.96640.966 2.201282.20128 151151 3.23.2
6363 46.74846.748 1.941631.94163 162162 3.43.4
6464 51.30151.301 1.779461.77946 175175 3.73.7
通过对表5～8任一表的数据和表2数据对比可知，对应衍射峰的位置都产生了不同程度的漂移，例如表2中2θ为4.183°的峰，其在表5、6、7、8中对应的2θ分别为3.901°(漂移程度为0.282°)、4.054°(漂移程度为0.129°)、3.973°(漂移程度为0.210°)、4.039°(漂移程度为0.144°)；或例如表2中2θ为16.789°的峰，其在表5、6、7、8中对应的2θ分别为16.512°(漂移程度为0.277°)、16.634°(漂移程度为0.155°)、16.535°(漂移程度为0.254°)、16.612°(漂移程度为0.177 °)；或例如表2中2θ为22.182°的峰，其在表5、6、7、8中对应的2θ分别为21.915 °(漂移程度为0.267°)、22.020°(漂移程度为0.162°)、21.933°(漂移程度为0.249°)、22.021°(漂移程度为0.161°)； By comparing the data in any of Tables 5 to 8 with the data in Table 2, it can be seen that the positions of the corresponding diffraction peaks have drifted to varying degrees. , and the corresponding 2θ in 8 are 3.901° (the degree of drift is 0.282°), 4.054° (the degree of drift is 0.129°), 3.973° (the degree of drift is 0.210°), and 4.039° (the degree of drift is 0.144°); or for example In Table 2, the 2θ is the peak of 16.789°, and its corresponding 2θ in Tables 5, 6, 7, and 8 are 16.512° (the degree of drift is 0.277°), 16.634° (the degree of drift is 0.155°), and 16.535° (the degree of drift is 0.155°). degree is 0.254 °), 16.612 ° (the degree of drift is 0.177 ° ); or for example, the peak of 2θ in Table 2 is 22.182 °, and its corresponding 2θ in Table 5, 6, 7, and 8 is respectively 21.915 ° (the degree of drift is 0.267°), 22.020° (the degree of drift is 0.162°), 21.933° (the degree of drift is 0.249°), 22.021° (the degree of drift is 0.161°);
尽管表5～8中与表2中对应衍射峰的位置有0.3°以内的漂移，但这些化合物皆是化合物N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型B。Although the positions of the corresponding diffraction peaks in Tables 5 to 8 and those in Table 2 have shifted within 0.3°, these compounds are all compounds N-(3'-(5-((6-acetyl-2,6-diazo Heterospiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl Form B of )-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide.
在本发明进一步优选的方案中，所述晶型为无水物。In a further preferred solution of the present invention, the crystal form is an anhydrate.
在本发明进一步优选的方案中，所述晶型为水合物，水的个数为0.2-3，优选0.2、0.5、1、1.5、2、2.5或3，更优选0.5、1、2或3。In a further preferred solution of the present invention, the crystal form is a hydrate, and the number of water is 0.2-3, preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3, more preferably 0.5, 1, 2 or 3 .
在本发明进一步优选的方案中，上述化合物晶型的制备方法，具体包括如下步骤：In a further preferred solution of the present invention, the preparation method of the crystal form of the above-mentioned compound specifically includes the following steps:
1)称取适量的自由碱，用良性溶剂在一定温度下溶解，温度优选0～50℃；1) Weigh an appropriate amount of free base and dissolve it in a benign solvent at a certain temperature, preferably 0-50°C;
2)任选地，向以上所得溶液中加入反溶剂，搅拌至固体析出；2) Optionally, add an anti-solvent to the solution obtained above, and stir until solids are precipitated;
3)任选地，搅拌、降温析晶得到目标产物；3) Optionally, stirring, cooling and crystallization to obtain the target product;
其中：in:
所述的良性溶剂选自甲醇、丙酮、甲酸乙酯、乙酸乙酯、乙腈、乙醇、88％丙酮、四氢呋喃、二氯甲烷、1,4-二氧六环、苯、甲苯、异丙醇、正丁醇、异丁 醇、N，N-二甲基甲酰胺、N，N-二甲基乙酰胺、正丙醇、叔丁醇、2-丁酮或3-戊酮中的一种或多种；优选甲酸乙酯、异丙醇或无水乙醇中的一种或多种；Described good solvent is selected from methanol, acetone, ethyl formate, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, isopropanol, One of n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol, 2-butanone or 3-pentanone or Multiple; preferably one or more of ethyl formate, isopropanol or absolute ethanol;
所述的反溶剂选自庚烷、环己烷、正己烷、正戊烷、水、甲基叔丁基醚、甲苯或异丙醚中的一种或多种；优选乙酸乙酯、甲基叔丁基醚或环己烷中的一种或多种。The anti-solvent is selected from one or more of heptane, cyclohexane, n-hexane, n-pentane, water, methyl tert-butyl ether, toluene or isopropyl ether; preferably ethyl acetate, methyl One or more of tert-butyl ether or cyclohexane.
在本发明进一步优选的方案中，上述化合物晶型的制备方法，具体包括如下步骤：In a further preferred solution of the present invention, the preparation method of the crystal form of the above-mentioned compound specifically includes the following steps:
1)称取适量的自由碱或其晶型，用不良溶剂在一定温度下打浆，温度优选0～50℃；1) Weighing an appropriate amount of free base or its crystal form, beating with a poor solvent at a certain temperature, preferably 0-50°C;
其中：in:
所述的不良溶剂选自庚烷、环己烷、正己烷、正戊烷、水、甲基叔丁基醚、甲苯或异丙醚中的一种或多种；优选乙酸乙酯、甲基叔丁基醚或环己烷中的一种或多种。Described poor solvent is selected from one or more in heptane, cyclohexane, n-hexane, n-pentane, water, methyl tert-butyl ether, toluene or isopropyl ether; Preferred ethyl acetate, methyl One or more of tert-butyl ether or cyclohexane.
本发明的目的还在于提供一种药物组合物，其含有治疗有效量的上述任一化合物的晶型，以及一种或多种药学上可接受的载体、稀释剂或赋形剂。The object of the present invention is also to provide a pharmaceutical composition, which contains a therapeutically effective amount of any crystal form of the above compound, and one or more pharmaceutically acceptable carriers, diluents or excipients.
本发明的目的还在于提供了上述任一化合物的晶型或上述药物组合物在制备PD-1/PD-L1抑制剂药物中的应用。The object of the present invention is also to provide the application of the crystal form of any of the above compounds or the above pharmaceutical composition in the preparation of PD-1/PD-L1 inhibitor drugs.
本发明的目的还在于提供了上述任一化合物的晶型或上述药物组合物在制备治疗疾病选自癌症、感染性疾病、自身免疫性疾病药物中的应用；其中所述的癌症选自皮肤癌、肺癌、泌尿系肿瘤、血液肿瘤、乳腺癌、胶质瘤、消化系统肿瘤、生殖系统肿瘤、淋巴瘤、神经系统肿瘤、脑瘤、头颈癌；所述的感染性疾病选自细菌感染、病毒感染；所述的自身免疫性疾病选自器官特异性自身免疫病、系统性自身免疫病，其中，所述的器官特异性自身免疫病包括慢性淋巴细胞性甲状腺炎、甲状腺功能亢进、胰岛素依赖型糖尿病、重症肌无力、溃疡性结肠炎、恶性贫血伴慢性萎缩性胃炎、肺出血肾炎综合征、原发性胆汁性肝硬化、多发性脑脊髓硬化症、急性特发性多神经炎，所述的系统性自身免疫病包括类风湿关节炎、系统性红斑狼疮、系统性血管炎、硬皮病、天疱疮、皮肌炎、混合性结缔组织病、自身免疫性溶血性贫血。The object of the present invention is also to provide the crystal form of any of the above compounds or the application of the above pharmaceutical composition in the preparation of medicines for treating diseases selected from cancer, infectious diseases, and autoimmune diseases; wherein the cancer is selected from skin cancer , lung cancer, urological tumor, blood tumor, breast cancer, glioma, digestive system tumor, reproductive system tumor, lymphoma, nervous system tumor, brain tumor, head and neck cancer; the infectious disease is selected from bacterial infection, viral infection; the autoimmune disease is selected from organ-specific autoimmune disease, systemic autoimmune disease, wherein, the organ-specific autoimmune disease includes chronic lymphocytic thyroiditis, hyperthyroidism, insulin-dependent Diabetes, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, pulmonary hemorrhage nephritic syndrome, primary biliary cirrhosis, multiple sclerosis, acute idiopathic polyneuritis, the Common systemic autoimmune diseases include rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, and autoimmune hemolytic anemia.
附图说明Description of drawings
图1为自由碱晶型A的XRPD图示。Fig. 1 is an XRPD diagram of free base crystal form A.
图2为自由碱晶型A的DSC图示。Fig. 2 is a DSC diagram of free base crystal form A.
图3为自由碱晶型B的XRPD图示。Figure 3 is an XRPD representation of free base Form B.
图4为自由碱晶型B的DSC图示。Fig. 4 is a DSC diagram of free base crystal form B.
图5为自由碱晶型C的XRPD图示。Figure 5 is an XRPD representation of free base Form C.
图6为自由碱晶型C的DSC图示。Figure 6 is a DSC diagram of free base Form C.
图7为自由碱晶型D的XRPD图示。Figure 7 is an XRPD representation of free base Form D.
图8为自由碱晶型D的DSC图示。Fig. 8 is a DSC diagram of free base crystal form D.
图9为自由碱晶型B批次一的XRPD图示。Fig. 9 is an XRPD diagram of batch 1 of free base crystal form B.
图10为自由碱晶型B批次二的XRPD图示。Fig. 10 is an XRPD diagram of batch 2 of free-base crystal form B.
图11为自由碱晶型B批次三的XRPD图示。Fig. 11 is an XRPD diagram of batch 3 of the free-base crystal form B.
图12为自由碱晶型B批次四的XRPD图示。Fig. 12 is an XRPD diagram of batch 4 of the free base crystal form B.
术语解释Terminology Explanation
除非有相反陈述，在说明书和权利要求书中使用的术语具有下述含义。Unless stated to the contrary, the terms used in the specification and claims have the following meanings.
术语“烷基”指饱和脂肪族烃基团，其为包含1至20个碳原子的直链或支链基团，优选含有1至8个碳原子的烷基，更优选1至6个碳原子的烷基，最更优选1至3个碳原子的烷基。非限制性实例包括甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基、正庚基、2-甲基己基、3-甲基己基、4-甲基己基、5-甲基己基、2,3-二甲基戊基、2,4-二甲基戊基、2,2-二甲基戊基、3,3-二甲基戊基、2-乙基戊基、3-乙基戊基、正辛基、2,3-二甲基己基、2,4-二甲基己基、2,5-二甲基己基、2,2-二甲基己基、3,3-二甲基己基、4,4-二甲基己基、2-乙基己基、3-乙基己基、4-乙基己基、2-甲基-2-乙基戊基、2-甲基-3-乙基戊基、正壬基、2-甲基-2-乙基己基、2-甲基-3-乙基己基、2,2-二乙基戊基、正癸基、3,3-二乙基己基、2,2-二乙基己基，及其各种支链异构体等。更优选的是含有1至6个碳原子的低级烷基，非限制性实施例包括甲基、乙基、正丙基、异丙基、正丁基、异丁基、叔丁基、仲丁基、正戊基、1,1-二甲基丙基、1,2-二甲基丙基、2,2-二甲基丙基、1-乙基丙基、2-甲基丁基、3-甲基丁基、正己基、1-乙基-2-甲基丙基、1,1,2-三甲基丙基、1,1-二甲基丁基、1,2-二甲基丁基、2,2-二甲基丁基、1,3-二甲基丁基、2-乙基丁基、2-甲基戊基、3-甲基戊基、4-甲基戊基、2,3-二甲基丁基等。烷基可以是取代的或非取代的，当被取代时，取代基可以在任何可使用的连接点上被取代，所述取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氧代基、羧基或羧酸酯基，本发明优选甲基、乙基、异丙基、叔丁基、卤代烷基、氘代烷基、烷氧基取代的烷基和羟基取代的烷基。The term "alkyl" refers to a saturated aliphatic hydrocarbon group which is a straight or branched chain group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 8 carbon atoms, more preferably 1 to 6 carbon atoms An alkyl group, most preferably an alkyl group of 1 to 3 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1 ,2-Dimethylpropyl, 2,2-Dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2- Methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3 -Dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2 -Methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethyl Pentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2 ,5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethylhexyl ylhexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethyl 2,2-diethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and various branched chain isomers. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl Base, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl Dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl group, 2,3-dimethylbutyl group, etc. Alkyl groups may be substituted or unsubstituted, and when substituted, substituents may be substituted at any available point of attachment, said substituents being preferably one or more of the following groups independently selected from alkyl radical, alkenyl, alkynyl, alkoxy, alkylthio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkane Oxygen, heterocycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo, carboxyl or carboxylate, preferably methyl, ethyl, isopropyl, tert-butyl, haloalkyl in the present invention , deuterated alkyl, alkoxy substituted alkyl and hydroxy substituted alkyl.
术语“亚烷基”是指烷基的一个氢原子进一步被取代，例如：“亚甲基”指-CH 2-、“亚乙基”指-(CH 2) 2-、“亚丙基”指-(CH 2) 3-、“亚丁基”指-(CH 2) 4-等。术语“烯基”指 由至少由两个碳原子和至少一个碳-碳双键组成的如上定义的烷基，例如乙烯基、1-丙烯基、2-丙烯基、1-、2-或3-丁烯基等。烯基可以是取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基。 The term "alkylene" means that one hydrogen atom of the alkyl group is further substituted, for example: "methylene" means -CH 2 -, "ethylene" means -(CH 2 ) 2 -, "propylene" refers to -(CH 2 ) 3 -, "butylene" refers to -(CH 2 ) 4 -, and the like. The term "alkenyl" means an alkyl group as defined above consisting of at least two carbon atoms and at least one carbon-carbon double bond, for example vinyl, 1-propenyl, 2-propenyl, 1-, 2- or 3- -butenyl etc. Alkenyl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycle Alkylthio.
术语“环烷基”指饱和或部分不饱和单环或多环环状烃取代基，环烷基环包含3至20个碳原子，优选包含3至10个碳原子，更优选包含3至8个碳原子，进一步优选包含3至6个碳原子。单环环烷基的非限制性实例包括环丙基、环丁基、环戊基、环戊烯基、环己基、环己烯基、环己二烯基、环庚基、环庚三烯基、环辛基等；多环环烷基包括螺环、稠环和桥环的环烷基，优选环丙基、环丁基、环戊基、环己基和环庚基。The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, the cycloalkyl ring containing 3 to 20 carbon atoms, preferably containing 3 to 10 carbon atoms, more preferably containing 3 to 8 carbon atoms, more preferably 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene group, cyclooctyl group, etc.; polycyclic cycloalkyl group includes spiro ring, fused ring and bridged ring cycloalkyl group, preferably cyclopropyl group, cyclobutyl group, cyclopentyl group, cyclohexyl group and cycloheptyl group.
所述环烷基环可以稠合于芳基、杂芳基或杂环烷基环上，其中与母体结构连接在一起的环为环烷基，非限制性实例包括茚满基、四氢萘基、苯并环庚烷基等。环烷基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氧代基、羧基或羧酸酯基。The cycloalkyl ring may be fused to an aryl, heteroaryl or heterocycloalkyl ring where the ring bonded to the parent structure is a cycloalkyl, non-limiting examples include indanyl, tetrahydronaphthalene base, benzocycloheptyl, etc. Cycloalkyl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, oxo, carboxyl or carboxylate.
术语“杂环基”指饱和或部分不饱和单环或多环环状烃取代基，其包含3至20个环原子，其中一个或多个环原子为选自氮、氧或S(O) m(其中m是整数0至2)的杂原子，但不包括-O-O-、-O-S-或-S-S-的环部分，其余环原子为碳。优选包含3至12个环原子，其中1～4个是杂原子；更优选包含3至10个环原子；进一步优选包含3至8个环原子。单环杂环基的非限制性实例包括噁嗪喃酮基、吡嗪酮基、吡啶酮基、吡咯烷基、四氢吡咯基、四氢吡咯酮基、氮杂环丁烷基、氧杂环丁烷基、氧杂环己烷基、咪唑烷基、四氢呋喃基、四氢噻吩基、二氢咪唑基、二氢呋喃基、二氢吡唑基、二氢吡咯基、哌啶基、哌啶酮基、哌嗪基、吗啉基、硫代吗啉基、高哌嗪基、四氢吡喃基和吡喃基等；优选噁嗪喃酮基、吡嗪酮基、吡啶酮基、吡咯烷基、四氢吡咯基、四氢吡咯酮基、氮杂环丁烷基、氧杂环丁烷基、四氢呋喃基、吡唑烷基、吗啉基、哌嗪基、哌啶基、哌啶酮基、四氢吡喃基和吡喃基；更优选噁嗪喃酮基、四氢呋喃基、异噁唑烷酮基、四氢吡咯基、四氢吡咯酮基、氮杂环丁烷基、氧杂环丁烷基、哌啶基、哌啶酮基和四氢吡喃基。多环杂环基包括螺环、稠环和桥环的杂环基；其中涉及到的螺环、稠环和桥环的杂环基任选与其他基团通过单键相连接，或者通过环上的任意两个或者两个以上的原子与其他环烷基、杂环基、芳基和杂芳基进一步并环连接。 The term "heterocyclyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent comprising 3 to 20 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (where m is an integer from 0 to 2), but excluding ring portions of -OO-, -OS- or -SS-, the remaining ring atoms being carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably contains 3 to 10 ring atoms; further preferably contains 3 to 8 ring atoms. Non-limiting examples of monocyclic heterocyclyl groups include oxazinone, pyrazinone, pyridinonyl, pyrrolidinyl, tetrahydropyrrolyl, tetrahydropyrrolidinyl, azetidinyl, oxa Cyclobutanyl, oxanyl, imidazolidinyl, tetrahydrofuranyl, tetrahydrothienyl, dihydroimidazolyl, dihydrofuryl, dihydropyrazolyl, dihydropyrrolyl, piperidinyl, piperyl Pyridonyl, piperazinyl, morpholinyl, thiomorpholinyl, homopiperazinyl, tetrahydropyranyl and pyranyl, etc.; preferably oxazinone, pyrazinone, pyridone, Pyrrolidinyl, tetrahydropyrrolyl, tetrahydropyrrolidinyl, azetidinyl, oxetanyl, tetrahydrofuranyl, pyrazolidinyl, morpholinyl, piperazinyl, piperidinyl, piperidine pyridonyl, tetrahydropyranyl and pyranyl; more preferably oxazinone, tetrahydrofuranyl, isoxazolidinyl, tetrahydropyrrolyl, tetrahydropyrrolidinyl, azetidinyl, Oxetanyl, piperidinyl, piperidinonyl and tetrahydropyranyl. Polycyclic heterocyclic groups include spiro rings, fused rings and bridged ring heterocyclic groups; the spiro rings, condensed rings and bridged ring heterocyclic groups involved are optionally connected to other groups through single bonds, or through rings Any two or more atoms on the ring are further linked with other cycloalkyl, heterocyclyl, aryl and heteroaryl groups.
所述杂环基环可以稠合于芳基、杂芳基或环烷基环上，其中与母体结构连接在一起的环为杂环基，其非限制性实例包括：The heterocyclyl ring may be fused to an aryl, heteroaryl, or cycloalkyl ring where the ring bonded to the parent structure is a heterocyclyl, non-limiting examples of which include:
Figure PCTCN2022102195-appb-000005
Figure PCTCN2022102195-appb-000005
杂环基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、氧代基、羧基或羧酸酯基。Heterocyclic groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alk Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, oxo, carboxyl or carboxylate.
术语“芳基”指具有共轭的π电子体系的6至14元全碳单环或稠合多环(也就是共享毗邻碳原子对的环)基团，优选为6至10元，更优选6至8元，例如苯基和萘基。更优选苯基。所述芳基环可以稠合于杂芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为芳基环，其非限制性实例包括：The term "aryl" refers to a 6 to 14 membered all-carbon monocyclic or fused polycyclic (that is, rings sharing adjacent pairs of carbon atoms) group having a conjugated π-electron system, preferably 6 to 10 membered, more preferably 6 to 8 membered, such as phenyl and naphthyl. Phenyl is more preferred. The aryl ring may be fused to a heteroaryl, heterocyclyl or cycloalkyl ring, where the ring bonded to the parent structure is an aryl ring, non-limiting examples of which include:
Figure PCTCN2022102195-appb-000006
Figure PCTCN2022102195-appb-000006
芳基可以是取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、羧基或羧酸酯基。Aryl groups may be substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio, Alkylamino, halogen, mercapto, hydroxy, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, heterocycle Alkylthio, carboxyl or carboxylate.
术语“杂芳基”指包含1至4个杂原子、5至14个环原子的杂芳族体系，其中杂原子选自氧、硫和氮。杂芳基优选为5至10元，更优选5至8元，最优选为5元或6元，例如咪唑基、呋喃基、噻吩基、噻唑基、吡唑基、噁唑基、吡咯基、三唑基、四唑基、吡啶基、嘧啶基、噻二唑、哒嗪基或吡嗪基等；优选三唑基、噻吩基、噻唑基、吡啶基、咪唑基、吡唑基、哒嗪基、吡嗪基或嘧啶基；更优选吡啶基、咪唑基、吡唑基、哒嗪基、吡嗪基或嘧啶基。所述杂芳基环可以稠合于芳基、杂环基或环烷基环上，其中与母体结构连接在一起的环为杂芳基环，其非限制性实例包括：The term "heteroaryl" refers to a heteroaromatic system comprising 1 to 4 heteroatoms, 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur and nitrogen. Heteroaryl is preferably 5 to 10 membered, more preferably 5 to 8 membered, most preferably 5 or 6 membered, such as imidazolyl, furyl, thienyl, thiazolyl, pyrazolyl, oxazolyl, pyrrolyl, Triazolyl, tetrazolyl, pyridyl, pyrimidyl, thiadiazole, pyridazinyl or pyrazinyl, etc.; preferably triazolyl, thienyl, thiazolyl, pyridyl, imidazolyl, pyrazolyl, pyridazine Base, pyrazinyl or pyrimidinyl; more preferably pyridyl, imidazolyl, pyrazolyl, pyridazinyl, pyrazinyl or pyrimidinyl. The heteroaryl ring may be fused to an aryl, heterocyclyl or cycloalkyl ring, wherein the ring bonded to the parent structure is a heteroaryl ring, non-limiting examples of which include:
Figure PCTCN2022102195-appb-000007
Figure PCTCN2022102195-appb-000007
Figure PCTCN2022102195-appb-000008
Figure PCTCN2022102195-appb-000008
杂芳基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、羧基或羧酸酯基。Heteroaryl groups may be optionally substituted or unsubstituted, and when substituted, the substituents are preferably one or more of the following groups independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkane Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, carboxyl or carboxylate.
术语“烷氧基”指-O-(烷基)和-O-(非取代的环烷基)，其中烷基的定义如上所述，优选含有1至8个碳原子的烷基，更优选1至6个碳原子的烷基，最更优选1至3个碳原子的烷基。烷氧基的非限制性实例包括：甲氧基、乙氧基、丙氧基、丁氧基、环丙氧基、环丁氧基、环戊氧基、环己氧基。烷氧基可以是任选取代的或非取代的，当被取代时，取代基优选为一个或多个以下基团，其独立地选自烷基、烯基、炔基、烷氧基、烷硫基、烷基氨基、卤素、巯基、羟基、硝基、氰基、环烷基、杂环烷基、芳基、杂芳基、环烷氧基、杂环烷氧基、环烷硫基、杂环烷硫基、羧基或羧酸酯基。The term "alkoxy" refers to -O-(alkyl) and -O-(unsubstituted cycloalkyl), wherein the definition of alkyl is as above, preferably alkyl containing 1 to 8 carbon atoms, more preferably An alkyl group of 1 to 6 carbon atoms, most preferably an alkyl group of 1 to 3 carbon atoms. Non-limiting examples of alkoxy include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. Alkoxy may be optionally substituted or unsubstituted, and when substituted, the substituent is preferably one or more of the following groups independently selected from the group consisting of alkyl, alkenyl, alkynyl, alkoxy, alkoxy Thio, alkylamino, halogen, mercapto, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , heterocycloalkylthio, carboxyl or carboxylate.
“卤代烷基”指被一个或多个卤素取代的烷基，其中烷基如上所定义。"Haloalkyl" means an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
“卤代烷氧基”指被一个或多个卤素取代的烷氧基，其中烷氧基如上所定义。"Haloalkoxy" means an alkoxy group substituted with one or more halogens, wherein alkoxy group is as defined above.
“羟烷基”指被羟基取代的烷基，其中烷基如上所定义。"Hydroxyalkyl" means an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.
“卤代烷基”指被一个或多个卤素取代的烷基，其中烷基如上所定义。"Haloalkyl" means an alkyl group substituted with one or more halogens, wherein alkyl is as defined above.
“卤代烷氧基”指被一个或多个卤素取代的烷氧基，其中烷氧基如上所定义。"Haloalkoxy" means an alkoxy group substituted with one or more halogens, wherein alkoxy group is as defined above.
“羟烷基”指被羟基取代的烷基，其中烷基如上所定义。"Hydroxyalkyl" means an alkyl group substituted with a hydroxy group, wherein alkyl is as defined above.
“羟基”指-OH基团。"Hydroxy" means an -OH group.
“卤素”指氟、氯、溴或碘。"Halogen" means fluorine, chlorine, bromine or iodine.
“氨基”指-NH 2。 "Amino" refers to -NH2 .
“氰基”指-CN。"Cyano" refers to -CN.
“硝基”指-NO 2。 "Nitro" refers to -NO2 .
“羧基”指-C(O)OH。"Carboxy" refers to -C(O)OH.
“THF”指四氢呋喃。"THF" means tetrahydrofuran.
“EtOAc”指乙酸乙酯。"EtOAc" means ethyl acetate.
“MeOH”指甲醇。"MeOH" means methanol.
“DMF”指N、N-二甲基甲酰胺。"DMF" refers to N,N-dimethylformamide.
“TFA”指三氟乙酸。"TFA" means trifluoroacetic acid.
“MeCN”指乙晴。"MeCN" refers to acetonitrile.
“DMA”指N,N-二甲基乙酰胺。"DMA" refers to N,N-dimethylacetamide.
“Et 2O”指乙醚。 " Et2O " means diethyl ether.
“DCE”指1,2二氯乙烷。"DCE" means 1,2 dichloroethane.
“DIPEA”指N，N-二异丙基乙胺。"DIPEA" means N,N-diisopropylethylamine.
“NBS”指N-溴代琥珀酰亚胺。"NBS" refers to N-bromosuccinimide.
“NIS”指N-碘代丁二酰亚胺。"NIS" refers to N-iodosuccinimide.
“Cbz-Cl”指氯甲酸苄酯。"Cbz-Cl" refers to benzyl chloroformate.
“Pd 2(dba) 3”指三(二亚苄基丙酮)二钯。 "Pd 2 (dba) 3 " refers to tris(dibenzylideneacetone)dipalladium.
“Dppf”指1,1’-双二苯基膦二茂铁。"Dppf" refers to 1,1'-bisdiphenylphosphinoferrocene.
“HATU”指2-(7-氧化苯并三氮唑)-N,N,N’,N’-四甲基脲六氟磷酸酯。"HATU" refers to 2-(7-benzotriazole oxide)-N,N,N',N'-tetramethyluronium hexafluorophosphate.
“KHMDS”指六甲基二硅基胺基钾。"KHMDS" refers to potassium hexamethyldisilazide.
“LiHMDS”指双三甲基硅基胺基锂。"LiHMDS" refers to lithium bistrimethylsilylamide.
“MeLi”指甲基锂。"MeLi" means methyllithium.
“n-BuLi”指正丁基锂。"n-BuLi" refers to n-butyllithium.
“NaBH(OAc) 3”指三乙酰氧基硼氢化钠。 "NaBH(OAc) 3 " refers to sodium triacetoxyborohydride.
“t-BuONO”指叔丁基亚硝酸酯。"t-BuONO" refers to t-butyl nitrite.
“EA”指乙酸乙酯。"EA" means ethyl acetate.
“PE”指石油醚。"PE" means petroleum ether.
“DCM”指二氯甲烷。"DCM" means dichloromethane.
“STAB”指三乙酰氧基硼氢化钠。"STAB" refers to sodium triacetoxyborohydride.
“Pd(dcypf)Cl 2”指二氯[1,1'-双(二环己基磷)二茂铁]钯。 "Pd(dcypf)Cl 2 " refers to dichloro[1,1'-bis(dicyclohexylphosphino)ferrocene]palladium.
“X选自A、B、或C”、“X选自A、B和C”、“X为A、B或C”、“X为A、B和C”等不同用语均表达了相同的意义，即表示X可以是A、B、C中的任意一种或几种。Different terms such as "X is selected from A, B, or C", "X is selected from A, B, and C", "X is A, B, or C", "X is A, B, and C" all express the same The meaning means that X can be any one or several of A, B, and C.
本发明所述的氢原子均可被其同位素氘所取代，本发明涉及的实施例化合物中的任一氢原子也均可被氘原子取代。The hydrogen atoms described in the present invention can be replaced by its isotope deuterium, and any hydrogen atom in the example compounds involved in the present invention can also be replaced by a deuterium atom.
“任选”或“任选地”意味着随后所描述的事件或环境可以但不必发生，该说明包括该事件或环境发生或不发生的场合。例如，“任选被烷基取代的杂环基团”意味着烷基可以但不必须存在，该说明包括杂环基团被烷基取代的情形和杂环基团不被烷基取代的情形。"Optional" or "optionally" means that the subsequently described event or circumstance can but need not occur, and that the description includes instances where the event or circumstance occurs or does not occur. For example, a "heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes cases where the heterocycle group is substituted with an alkyl group and cases where the heterocycle group is not substituted with an alkyl group .
“取代的”指基团中的一个或多个氢原子，优选为最多5个，更优选为1～3个氢原子彼此独立地被相应数目的取代基取代。不言而喻，取代基仅处在它们的可能的化学位置，本领域技术人员能够在不付出过多努力的情况下确定(通过实验或理论)可能或不可能的取代。例如，具有游离氢的氨基或羟基与具有不饱和(如烯属)键的碳原子结合时可能是不稳定的。"Substituted" means that one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms are independently substituted by the corresponding number of substituents. It goes without saying that substituents are only in their possible chemical positions and that a person skilled in the art can determine (by experiment or theory) possible or impossible substitutions without undue effort. For example, an amino or hydroxyl group with free hydrogen may be unstable when bonded to a carbon atom with an unsaturated (eg, ethylenic) bond.
“药物组合物”表示含有一种或多种本文所述化合物或其生理学上/可药用的盐或前体药物与其他化学组分的混合物，以及其他组分例如生理学/可药用的载体和赋形剂。药物组合物的目的是促进对生物体的给药，利于活性成分的吸收进而发挥生物活性。"Pharmaceutical composition" means a mixture containing one or more compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, and other components such as a physiologically/pharmaceutically acceptable carrier and excipients. The purpose of the pharmaceutical composition is to promote the administration to the organism, facilitate the absorption of the active ingredient and thus exert biological activity.
具体实施方式detailed description
以下结合实施例进一步描述本发明，但这些实施例并非限制着本发明的范围。The present invention is further described below in conjunction with examples, but these examples do not limit the scope of the present invention.
实施例Example
本发明的化合物结构是通过核磁共振(NMR)或/和液质联用色谱(LC-MS)来确定的。NMR化学位移(δ)以百万分之一(ppm)的单位给出。NMR的测定是用Bruker AVANCE-400核磁仪，测定溶剂为氘代二甲基亚砜(DMSO-d 6)，氘代甲醇(CD 3OD)和氘代氯仿(CDCl 3)，内标为四甲基硅烷(TMS)。 The structures of the compounds of the present invention are determined by nuclear magnetic resonance (NMR) or/and liquid chromatography-mass chromatography (LC-MS). NMR chemical shifts (δ) are given in parts per million (ppm). The determination of NMR is to use Bruker AVANCE-400 nuclear magnetic apparatus, and the determination solvent is deuterated dimethyl sulfoxide (DMSO-d 6 ), deuterated methanol (CD 3 OD) and deuterated chloroform (CDCl 3 ), and the internal standard is four Methylsilane (TMS).
液质联用色谱LC-MS的测定用Agilent 1200 Infinity Series质谱仪。HPLC的测定使用安捷伦1200DAD高压液相色谱仪(Sunfire C18 150×4.6mm色谱柱)和Waters 2695-2996高压液相色谱仪(Gimini C18 150×4.6mm色谱柱)。 Agilent 1200 Infinity Series mass spectrometer was used for liquid chromatography-mass chromatography LC-MS determination. The determination of HPLC used Agilent 1200DAD high pressure liquid chromatography (Sunfire C18 150×4.6mm column) and Waters 2695-2996 high pressure liquid chromatography (Gimini C18 150×4.6mm column).
薄层层析硅胶板使用烟台黄海HSGF254或青岛GF254硅胶板，TLC采用的规格是0.15mm～0.20mm，薄层层析分离纯化产品采用的规格是0.4mm～0.5mm。柱层析一般使用烟台黄海硅胶200～300目硅胶为载体。Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates are used for thin-layer chromatography silica gel plates. The specifications used for TLC are 0.15mm-0.20mm, and the specifications used for thin-layer chromatography separation and purification products are 0.4mm-0.5mm. Column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.
本发明实施例中的起始原料是已知的并且可以在市场上买到，或者可以采用或按照本领域已知的方法来合成。The starting materials in the examples of the present invention are known and commercially available, or can be synthesized using or following methods known in the art.
在无特殊说明的情况下，本发明的所有反应均在连续的磁力搅拌下，在干燥氮气或氩气氛下进行，溶剂为干燥溶剂，反应温度单位为摄氏度。Unless otherwise specified, all the reactions in the present invention are carried out under a dry nitrogen or argon atmosphere under continuous magnetic stirring, the solvent is a dry solvent, and the unit of the reaction temperature is Celsius.
实施例1Example 1
N-(2,2'-二氯-3'-(6-甲氧基-5-((噁丁环-3-基氨基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(6-methoxy-5-((oxetan-3-ylamino)methyl)pyridin-2-yl)-[1,1'- Biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000009
Figure PCTCN2022102195-appb-000009
第一步：6-(3-溴-2-氯苯基)-2-甲氧基尼古丁醛的制备The first step: the preparation of 6-(3-bromo-2-chlorophenyl)-2-methoxynicotinaldehyde
Figure PCTCN2022102195-appb-000010
Figure PCTCN2022102195-appb-000010
6-氯-2-甲氧基尼古丁醛(400mg,2.34mmol)、(3-溴-2-氯苯基)硼酸(600mg, 2.57mmol)，Pd(PPh 3) 4(266mg,0.23mmol)和碳酸钾(646mg,4.68mmol)溶于二氧六环(20mL)和水(2mL)的混合溶剂中，抽换气，氮气保护后，加热至95℃搅拌过夜。反应完全后，将反应液冷却，过滤，滤液浓缩后，残留物经快速硅胶柱层析(PE:EA＝4:1)分离得到标题化合物(600mg,78％)。 6-Chloro-2-methoxynicotinaldehyde (400mg, 2.34mmol), (3-bromo-2-chlorophenyl)boronic acid (600mg, 2.57mmol), Pd(PPh 3 ) 4 (266mg, 0.23mmol) and Potassium carbonate (646mg, 4.68mmol) was dissolved in a mixed solvent of dioxane (20mL) and water (2mL), ventilated, and heated to 95°C and stirred overnight under nitrogen protection. After the reaction was complete, the reaction solution was cooled, filtered, and the filtrate was concentrated. The residue was separated by flash silica gel column chromatography (PE:EA=4:1) to obtain the title compound (600 mg, 78%).
MS m/z(ESI):326.7[M+H] +. MS m/z(ESI):326.7[M+H] + .
第二步：6-(2-氯-3-(4,4,5,5-四甲基-1,3,2-二噁硼戊环-2-基)苯基)-2-甲氧基尼古丁醛的制备The second step: 6-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)-2-methoxy Preparation of Nicotinaldehyde
Figure PCTCN2022102195-appb-000011
Figure PCTCN2022102195-appb-000011
将6-(3-溴-2-氯苯基)-2-甲氧基尼古丁醛(600mg,1.83mmol)，片呐醇联硼酸酯(607mg,2.39mol)，Pd(dppf)Cl 2DCM络合物(149mg,0.18mmol)和乙酸钾(538mg,5.49mmol)溶于二氧六环(17mL)中，抽换气，氮气保护，于95℃加热搅拌过夜。反应完全后，将反应液冷却，浓缩。残留物经快速硅胶柱层析(PE:EA＝3:1)分离得到标题化合物(600mg,87％)。 6-(3-Bromo-2-chlorophenyl)-2-methoxynicotinaldehyde (600mg, 1.83mmol), pinacol bis-boronate (607mg, 2.39mol), Pd(dppf)Cl 2 DCM The complex (149mg, 0.18mmol) and potassium acetate (538mg, 5.49mmol) were dissolved in dioxane (17mL), gas exchanged, nitrogen protection, heated and stirred at 95°C overnight. After the reaction was complete, the reaction solution was cooled and concentrated. The residue was separated by flash silica gel column chromatography (PE:EA=3:1) to obtain the title compound (600 mg, 87%).
MS m/z(ESI):375.7[M+H] +. MS m/z(ESI):375.7[M+H] + .
第三步：N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备The third step: N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl )-1,5-Dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000012
Figure PCTCN2022102195-appb-000012
将N-(3-溴-2-氯苯基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(170mg,0.444mmol)，6-(2-氯-3-(4,4,5,5-四甲基-1,3,2-二噁硼戊环-2-基)苯基)-2-甲氧基尼古丁醛(216mg,0.577mol),Pd(dppf)Cl 2DCM络合物(73mg,0.09mmol)和碳酸铯(360mg,1.11mmol)溶于二氧六环(10mL)和水(2mL)的中混合溶剂中，抽换气，氮气保护，于95℃加热搅拌过夜。反应完全后，将反应液冷却，浓缩。残留物经反相分离得到标题化合物(200mg,82％)。 N-(3-bromo-2-chlorophenyl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-methyl Amide (170mg, 0.444mmol), 6-(2-chloro-3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)- 2-Methoxynicotinaldehyde (216 mg, 0.577 mol), Pd(dppf)Cl 2 DCM complex (73 mg, 0.09 mmol) and cesium carbonate (360 mg, 1.11 mmol) were dissolved in dioxane (10 mL) and water (2 mL) in a mixed solvent, pumped for gas, nitrogen protection, heated and stirred at 95°C overnight. After the reaction was complete, the reaction solution was cooled and concentrated. The residue was separated by reverse phase to obtain the title compound (200 mg, 82%).
MS m/z(ESI):550.7[M+H] +. MS m/z(ESI):550.7[M+H] + .
第四步：N-(2,2'-二氯-3'-(6-甲氧基-5-((噁丁环-3-基氨基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备The fourth step: N-(2,2'-dichloro-3'-(6-methoxy-5-((oxetan-3-ylamino)methyl)pyridin-2-yl)-[1 ,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide preparation of
Figure PCTCN2022102195-appb-000013
Figure PCTCN2022102195-appb-000013
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(100mg,0.181mmol)和噁丁环-3-胺(52mg,0.726mmol)溶解于甲醇(3mL)中，加入4滴乙酸后，于室温下搅拌3小时。加入氰基硼氢化钠(22mg,0.362mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过反相纯化得到标题化合物(10.3mg,9％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (100mg, 0.181mmol) and oxetane-3-amine ( 52mg, 0.726mmol) was dissolved in methanol (3mL), after adding 4 drops of acetic acid, stirred at room temperature for 3 hours. After adding sodium cyanoborohydride (22mg, 0.362mmol), stirring was continued at room temperature overnight. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by reverse phase to obtain the title compound (10.3 mg, 9%).
MS m/z(ESI):607.2[M+H] +. MS m/z(ESI):607.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.35(d,J＝8.3Hz,1H),7.79(d,J＝7.5Hz,1H),7.67(d,J＝7.6Hz,1H),7.51(dt,J＝25.0,7.7Hz,2H),7.41(d,J＝7.6Hz,1H),7.26(d,J＝7.5Hz,1H),7.18(d,J＝7.6Hz,1H),4.59(t,J＝6.5Hz,2H),4.32(t,J＝6.2Hz,2H),3.91(d,J＝8.4Hz,7H),3.63(s,2H),3.37(s,2H),2.69(s,4H),2.38(s,3H). 1 H NMR (400MHz, DMSO-d 6 )δ9.91(s, 1H), 8.35(d, J=8.3Hz, 1H), 7.79(d, J=7.5Hz, 1H), 7.67(d, J= 7.6Hz, 1H), 7.51(dt, J=25.0, 7.7Hz, 2H), 7.41(d, J=7.6Hz, 1H), 7.26(d, J=7.5Hz, 1H), 7.18(d, J= 7.6Hz, 1H), 4.59(t, J=6.5Hz, 2H), 4.32(t, J=6.2Hz, 2H), 3.91(d, J=8.4Hz, 7H), 3.63(s, 2H), 3.37 (s,2H),2.69(s,4H),2.38(s,3H).
实施例2Example 2
N-(2,2'-二氯-3'-(5-((3-氟吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((3-fluoroazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000014
Figure PCTCN2022102195-appb-000014
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(55.1mg,0.1mmol)和3-氟吖丁啶盐酸(33.3mg,0.3mmol)溶解于甲醇(3mL)中，加入DIPEA中和后，加入3滴乙酸，于室温下搅拌3小时。加入氰基硼氢化钠(18.6mg,0.3mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过反相纯化得到标题化合物(14mg,23％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (55.1mg, 0.1mmol) and 3-fluoroazetidine hydrochloride (33.3 mg, 0.3 mmol) was dissolved in methanol (3 mL), neutralized by adding DIPEA, then added 3 drops of acetic acid, and stirred at room temperature for 3 hours. After adding sodium cyanoborohydride (18.6 mg, 0.3 mmol), the stirring was continued at room temperature overnight. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by reverse phase to obtain the title compound (14 mg, 23%).
MS m/z(ESI):609.3[M+H] +. MS m/z(ESI):609.3[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.35(d,J＝8.4Hz,1H),7.69(dd,J＝12.5,7.5Hz,2H),7.51(dt,J＝23.9,7.9Hz,2H),7.41(d,J＝7.6Hz,1H),7.27(d,J＝7.5Hz,1H),7.18(d,J＝7.6Hz,1H),5.27(d,J＝7.3Hz,1H),5.12(d,J＝7.1Hz, 1H),3.94–3.85(m,5H),3.66(d,J＝8.0Hz,4H),3.39(s,2H),3.30–3.22(m,1H),3.20(s,1H),2.70(s,4H),2.39(s,3H). 1 H NMR (400MHz, DMSO-d 6 )δ9.91(s, 1H), 8.35(d, J=8.4Hz, 1H), 7.69(dd, J=12.5, 7.5Hz, 2H), 7.51(dt, J=23.9,7.9Hz,2H),7.41(d,J=7.6Hz,1H),7.27(d,J=7.5Hz,1H),7.18(d,J=7.6Hz,1H),5.27(d, J＝7.3Hz, 1H), 5.12(d, J＝7.1Hz, 1H), 3.94–3.85(m, 5H), 3.66(d, J＝8.0Hz, 4H), 3.39(s, 2H), 3.30– 3.22(m,1H),3.20(s,1H),2.70(s,4H),2.39(s,3H).
实施例3Example 3
1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-基乙酸酯1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-carbobalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine- 3-yl acetate
Figure PCTCN2022102195-appb-000015
Figure PCTCN2022102195-appb-000015
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(73mg,0.106mmol)和吖丁啶-3-基乙酸酯三氟乙酸盐(61mg,0.266mmol)溶解于甲醇(4mL)中，加入DIPEA中和后，加入4滴乙酸，于室温下搅拌3小时。加入氰基硼氢化钠(16mg,0.266mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过快速硅胶柱层析(DCM:MeOH＝10:1)纯化得到粗产品后，经过反相纯化得到标题化合物(9.9mg,15％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-Dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (73mg, 0.106mmol) and azetidin-3-ylethyl Ester trifluoroacetate (61 mg, 0.266 mmol) was dissolved in methanol (4 mL), neutralized by adding DIPEA, then added 4 drops of acetic acid, and stirred at room temperature for 3 hours. After adding sodium cyanoborohydride (16mg, 0.266mmol), stirring was continued at room temperature overnight. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by flash silica gel column chromatography (DCM:MeOH=10:1) to obtain a crude product, and then purified by reverse phase to obtain the title compound (9.9 mg, 15%).
MS m/z(ESI):649.3[M+H] +. MS m/z(ESI):649.3[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.38–8.31(m,1H),7.74–7.64(m,2H),7.54(t,J＝7.6Hz,1H),7.48(t,J＝7.9Hz,1H),7.41(dd,J＝7.5,1.7Hz,1H),7.26(d,J＝7.5Hz,1H),7.18(dd,J＝7.6,1.6Hz,1H),4.96(t,J＝5.8Hz,1H),3.91(d,J＝3.9Hz,6H),3.70–3.60(m,4H),3.37(s,2H),3.11(dd,J＝8.5,5.4Hz,2H),2.69(s,4H),2.38(s,3H),2.03(s,3H). 1 H NMR (400MHz,DMSO-d 6 )δ9.91(s,1H),8.38–8.31(m,1H),7.74–7.64(m,2H),7.54(t,J=7.6Hz,1H), 7.48(t, J=7.9Hz, 1H), 7.41(dd, J=7.5, 1.7Hz, 1H), 7.26(d, J=7.5Hz, 1H), 7.18(dd, J=7.6, 1.6Hz, 1H ), 4.96(t, J＝5.8Hz, 1H), 3.91(d, J＝3.9Hz, 6H), 3.70–3.60(m, 4H), 3.37(s, 2H), 3.11(dd, J＝8.5, 5.4Hz,2H),2.69(s,4H),2.38(s,3H),2.03(s,3H).
实施例4Example 4
N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000016
Figure PCTCN2022102195-appb-000016
Figure PCTCN2022102195-appb-000017
Figure PCTCN2022102195-appb-000017
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(115mg,0.208mmol)和1-(2,6-二氮杂螺[3.3]庚烷-2-基)乙烷-1-酮盐酸(110mg,0.625mmol)溶解于甲醇(4mL)中，加入DIPEA中和后，加入8滴乙酸，于室温下搅拌3小时。加入氰基硼氢化钠(38.8mg,0.625mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过反相纯化得到标题化合物(23.4mg,17％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-Dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (115mg, 0.208mmol) and 1-(2,6-di Azaspiro[3.3]heptan-2-yl)ethan-1-one hydrochloride (110mg, 0.625mmol) was dissolved in methanol (4mL), neutralized by adding DIPEA, added 8 drops of acetic acid, stirred at room temperature for 3 Hour. After adding sodium cyanoborohydride (38.8 mg, 0.625 mmol), the stirring was continued at room temperature overnight. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by reverse phase to obtain the title compound (23.4 mg, 17%).
MS m/z(ESI):674.3[M+H] +. MS m/z(ESI):674.3[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.34(dd,J＝8.3,1.5Hz,1H),7.75-7.64(m,2H),7.52(dt,J＝24.3,7.8Hz,2H),7.41(dd,J＝7.6,1.7Hz,1H),7.28(d,J＝7.4Hz,1H),7.18(dd,J＝7.6,1.6Hz,1H),4.19(s,2H),3.91(d,J＝4.1Hz,7H),3.53(d,J＝65.9Hz,9H),2.74(d,J＝20.1Hz,4H),2.43(s,3H),1.72(s,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.91(s, 1H), 8.34(dd, J=8.3, 1.5Hz, 1H), 7.75-7.64(m, 2H), 7.52(dt, J=24.3 ,7.8Hz,2H),7.41(dd,J=7.6,1.7Hz,1H),7.28(d,J=7.4Hz,1H),7.18(dd,J=7.6,1.6Hz,1H),4.19(s ,2H),3.91(d,J=4.1Hz,7H),3.53(d,J=65.9Hz,9H),2.74(d,J=20.1Hz,4H),2.43(s,3H),1.72(s ,3H).
实施例5Example 5
N-(3'-(5-((2-氧杂-6-氮杂螺[3.3]庚烷-6-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((2-Oxa-6-azaspiro[3.3]heptane-6-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000018
Figure PCTCN2022102195-appb-000018
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(50mg,0.091mmol)和2-氧杂-6-氮杂螺[3.3]庚烷(27mg,0.272mmol)溶解于甲醇(3mL)中，加入2滴乙酸，于室温下搅拌2小时。加入氰基硼氢化钠(14mg,0.227mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过反相纯化得到标题化合物(10mg,18％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (50mg, 0.091mmol) and 2-oxa-6-azepam Spiro[3.3]heptane (27mg, 0.272mmol) was dissolved in methanol (3mL), 2 drops of acetic acid was added, and stirred at room temperature for 2 hours. After adding sodium cyanoborohydride (14 mg, 0.227 mmol), the stirring was continued overnight at room temperature. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by reverse phase to obtain the title compound (10 mg, 18%).
MS m/z(ESI):633.3[M+H] +. MS m/z(ESI):633.3[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.35(d,J＝8.2Hz,1H),7.67(t,J＝5.8Hz,2H),7.51(dt,J＝23.1,7.8Hz,2H),7.40(d,J＝7.5Hz,1H),7.25(d,J＝7.5 Hz,1H),7.18(d,J＝7.7Hz,1H),4.62(s,4H),3.90(s,6H),3.51(s,2H),3.38(d,J＝6.6Hz,6H),2.69(s,4H),2.38(s,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.91(s, 1H), 8.35(d, J=8.2Hz, 1H), 7.67(t, J=5.8Hz, 2H), 7.51(dt, J= 23.1,7.8Hz,2H),7.40(d,J=7.5Hz,1H),7.25(d,J=7.5Hz,1H),7.18(d,J=7.7Hz,1H),4.62(s,4H) ,3.90(s,6H),3.51(s,2H),3.38(d,J=6.6Hz,6H),2.69(s,4H),2.38(s,3H).
实施例6Example 6
N-(3'-(5-((5-氧杂-2-氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((5-oxa-2-azaspiro[3.4]octane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000019
Figure PCTCN2022102195-appb-000019
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(76mg,0.134mmol)和5-氧杂-2-氮杂螺[3.4]辛烷盐酸盐(31mg,0.206mmol)溶解于甲醇(4mL)中，加入DIPEA中和后，加入4滴乙酸，于室温下搅拌2小时。加入氰基硼氢化钠(17mg,0.274mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过反相纯化得到标题化合物(6.9mg,8％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (76mg, 0.134mmol) and 5-oxa-2-azepam Spiro[3.4]octane hydrochloride (31 mg, 0.206 mmol) was dissolved in methanol (4 mL), neutralized by adding DIPEA, then added 4 drops of acetic acid, and stirred at room temperature for 2 hours. After adding sodium cyanoborohydride (17mg, 0.274mmol), stirring was continued at room temperature overnight. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by reverse phase to obtain the title compound (6.9 mg, 8%).
MS m/z(ESI):647.3[M+H] +. MS m/z(ESI):647.3[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.35(d,J＝8.2Hz,1H),7.69(dd,J＝15.2,7.6Hz,2H),7.51(dt,J＝23.6,7.8Hz,2H),7.40(d,J＝7.5Hz,1H),7.26(d,J＝7.5Hz,1H),7.18(d,J＝7.6Hz,1H),3.90(s,6H),3.67(t,J＝6.8Hz,2H),3.59(s,2H),3.37(s,4H),3.09(d,J＝6.9Hz,2H),2.69(s,4H),2.38(s,3H),2.04(t,J＝7.2Hz,2H),1.82(q,J＝6.7Hz,2H). 1 H NMR (400MHz, DMSO-d 6 )δ9.91(s, 1H), 8.35(d, J=8.2Hz, 1H), 7.69(dd, J=15.2, 7.6Hz, 2H), 7.51(dt, J=23.6,7.8Hz,2H),7.40(d,J=7.5Hz,1H),7.26(d,J=7.5Hz,1H),7.18(d,J=7.6Hz,1H),3.90(s, 6H), 3.67(t, J=6.8Hz, 2H), 3.59(s, 2H), 3.37(s, 4H), 3.09(d, J=6.9Hz, 2H), 2.69(s, 4H), 2.38( s,3H),2.04(t,J=7.2Hz,2H),1.82(q,J=6.7Hz,2H).
实施例7Example 7
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-羰基-2,5-二氮杂螺[3.4]辛烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(6-methoxy-5-((6-carbonyl-2,5-diazaspiro[3.4]octane-2-yl)methyl) Pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5 -c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000020
Figure PCTCN2022102195-appb-000020
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-羰基-2,5-二氮杂螺[3.4]辛烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰 胺的制备参照实施例1。N-(2,2'-dichloro-3'-(6-methoxy-5-((6-carbonyl-2,5-diazaspiro[3.4]octane-2-yl)methyl) Pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5 -c] The preparation of pyridine-2-carboxamide refers to Example 1.
MS m/z(ESI):660.2[M+H] +. MS m/z(ESI):660.2[M+H] + .
实施例8Example 8
N-(2,2'-二氯-3'-(5-((3-羟基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((3-hydroxyazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000021
Figure PCTCN2022102195-appb-000021
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(30mg,0.055mmol)和吖丁啶-3-醇盐酸盐(24mg,0.217mmol)溶解于甲醇(3mL)中，加入DIPEA中和后，加入3滴乙酸，于室温下搅拌3小时。加入氰基硼氢化钠(6.7mg,0.109mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过反相纯化得到标题化合物(5.3mg,16％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-Dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (30mg, 0.055mmol) and azetidine-3-alcohol The acid salt (24mg, 0.217mmol) was dissolved in methanol (3mL), neutralized by adding DIPEA, then added 3 drops of acetic acid, and stirred at room temperature for 3 hours. After adding sodium cyanoborohydride (6.7 mg, 0.109 mmol), the stirring was continued at room temperature overnight. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by reverse phase to obtain the title compound (5.3 mg, 16%).
MS m/z(ESI):607.2[M+H] +. MS m/z(ESI):607.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.35(d,J＝8.3Hz,1H),7.68(t,J＝7.1Hz,2H),7.51(dt,J＝23.4,7.8Hz,2H),7.40(d,J＝7.8Hz,1H),7.26(d,J＝7.5Hz,1H),7.18(d,J＝7.7Hz,1H),5.33(s,1H),4.26–4.18(m,1H),3.90(d,J＝3.1Hz,5H),3.58(d,J＝11.1Hz,4H),3.37(s,2H),2.84(t,J＝6.7Hz,2H),2.69(s,4H),2.38(s,3H),2.05–1.97(m,1H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.91(s, 1H), 8.35(d, J=8.3Hz, 1H), 7.68(t, J=7.1Hz, 2H), 7.51(dt, J= 23.4,7.8Hz,2H),7.40(d,J=7.8Hz,1H),7.26(d,J=7.5Hz,1H),7.18(d,J=7.7Hz,1H),5.33(s,1H) ,4.26–4.18(m,1H),3.90(d,J=3.1Hz,5H),3.58(d,J=11.1Hz,4H),3.37(s,2H),2.84(t,J=6.7Hz, 2H), 2.69(s,4H), 2.38(s,3H), 2.05–1.97(m,1H).
实施例9Example 9
N-(2,2'-二氯-3'-(5-((3-羟基-3-甲基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((3-hydroxy-3-methylazetidin-1-yl)methyl)-6-methoxypyridin-2-yl) -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 -Formamide
Figure PCTCN2022102195-appb-000022
Figure PCTCN2022102195-appb-000022
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(30mg,0.055mmol)和3-甲基吖丁啶-3-醇盐酸(27mg,0.218mmol)溶解于甲醇(3mL)中，加入DIPEA中和后，加入3滴乙酸，于室温下搅拌3小时。加入氰基硼氢化钠(6.8mg,0.109mmol)后，继续于室温搅拌过夜。反应完全后，将反应液浓缩，残留物经过反相纯化得到标题化合物(6.4mg,19％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-Dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (30mg, 0.055mmol) and 3-methylazetidine- 3-Alcohol hydrochloric acid (27 mg, 0.218 mmol) was dissolved in methanol (3 mL), neutralized by adding DIPEA, then added 3 drops of acetic acid, and stirred at room temperature for 3 hours. After adding sodium cyanoborohydride (6.8 mg, 0.109 mmol), the stirring was continued overnight at room temperature. After the reaction was complete, the reaction solution was concentrated, and the residue was purified by reverse phase to obtain the title compound (6.4 mg, 19%).
MS m/z(ESI):621.2[M+H] +. MS m/z(ESI):621.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.91(s,1H),8.35(d,J＝8.4Hz,1H),7.69(t,J＝9.3Hz,2H),7.51(dt,J＝23.5,7.8Hz,2H),7.40(d,J＝7.6Hz,1H),7.27(d,J＝7.4Hz,1H),7.18(d,J＝7.7Hz,1H),3.93–3.86(m,6H),3.59(s,2H),3.37(s,2H),3.29(d,J＝6.6Hz,2H),2.97(d,J＝6.6Hz,2H),2.69(s,4H),2.39(d,J＝3.4Hz,3H),1.39(s,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.91(s, 1H), 8.35(d, J=8.4Hz, 1H), 7.69(t, J=9.3Hz, 2H), 7.51(dt, J= 23.5,7.8Hz,2H),7.40(d,J=7.6Hz,1H),7.27(d,J=7.4Hz,1H),7.18(d,J=7.7Hz,1H),3.93–3.86(m, 6H), 3.59(s, 2H), 3.37(s, 2H), 3.29(d, J=6.6Hz, 2H), 2.97(d, J=6.6Hz, 2H), 2.69(s, 4H), 2.39( d,J=3.4Hz,3H),1.39(s,3H).
实施例10Example 10
1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺基)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-羧酸1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-Carboxamido)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine-3-carboxylic acid
Figure PCTCN2022102195-appb-000023
Figure PCTCN2022102195-appb-000023
1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺基)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-羧酸的制备参照实施例1。1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine The preparation of -2-formamido)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine-3-carboxylic acid refers to the implementation example 1.
MS m/z(ESI):635.2[M+H] +. MS m/z(ESI):635.2[M+H] + .
实施例11Example 11
(S)-1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-2-羧酸(S)-1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5 -c]pyridine-2-carboxalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl) Azetidine-2-carboxylic acid
Figure PCTCN2022102195-appb-000024
Figure PCTCN2022102195-appb-000024
将(2S)-吖丁啶-2-羧酸(11.02mg,109.00μmol)加到N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(50mg,90.84μmol)的AcOH(10.00μL)、DMF(0.5mL)和MeOH(0.5mL)的混合溶液中，室温搅拌2小时，再加入氰基硼氢化钠(95.83mg,454.19μmol)，室温搅拌16小时。反应液用prep-HPLC分离纯化得到标题化合物为白色固体Add (2S)-azetidine-2-carboxylic acid (11.02 mg, 109.00 μmol) to N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] In a mixed solution of pyridine-2-carboxamide (50mg, 90.84μmol) in AcOH (10.00μL), DMF (0.5mL) and MeOH (0.5mL), stir at room temperature for 2 hours, then add sodium cyanoborohydride (95.83mg ,454.19μmol), stirred at room temperature for 16 hours. The reaction solution was separated and purified by prep-HPLC to obtain the title compound as a white solid
(16.5mg,26％)。(16.5 mg, 26%).
MS m/z(ESI):635.2[M+H] +. MS m/z(ESI):635.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.90(s,1H),8.35(dd,J＝8.3,1.6Hz,1H),7.79(d,J＝7.5Hz,1H),7.68(dd,J＝7.7,1.7Hz,1H),7.54(t,J＝7.6Hz,1H),7.48(t,J＝8.0Hz,1H),7.41(dd,J＝7.6,1.7Hz,1H),7.28(d,J＝7.5Hz,1H),7.18(d,J＝7.6Hz,1H),3.94-3.87(m,8H),3.76-3.70(m,1H),3.44-3.41(m,1H),3.37(s,2H),3.13-3.08(m,1H),2.71-2.64(m,4H),2.39(s,3H),2.26-2.16(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ9.90(s, 1H), 8.35(dd, J=8.3, 1.6Hz, 1H), 7.79(d, J=7.5Hz, 1H), 7.68(dd, J=7.7,1.7Hz,1H),7.54(t,J=7.6Hz,1H),7.48(t,J=8.0Hz,1H),7.41(dd,J=7.6,1.7Hz,1H),7.28( d,J=7.5Hz,1H),7.18(d,J=7.6Hz,1H),3.94-3.87(m,8H),3.76-3.70(m,1H),3.44-3.41(m,1H),3.37 (s,2H),3.13-3.08(m,1H),2.71-2.64(m,4H),2.39(s,3H),2.26-2.16(m,2H).
实施例12Example 12
(S)-N-(2,2'-二氯-3'-(5-((2-(羟甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(S)-N-(2,2'-dichloro-3'-(5-((2-(hydroxymethyl)azetidin-1-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000025
Figure PCTCN2022102195-appb-000025
-20℃下将氯甲酸异丁酯(3.01mg,22.01μmol)的THF(0.1mL)溶液滴加到(S)-1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-2-羧酸甲酸盐(10mg,14.67μmol)和DIPEA(6.64mg,51.35μmol)的THF(2mL)溶液中，在此温度下反应10分钟，然后加入NaBH 4(0.83mg,22.01μmol)的水(0.5mL)溶液，缓慢升至室温搅拌1小时。反应液用prep-HPLC分离纯化得到标题化合物为灰白色固体(2.3mg,23％)。 A solution of isobutyl chloroformate (3.01 mg, 22.01 μmol) in THF (0.1 mL) was added dropwise to (S)-1-((6-(2,2'-dichloro-3'-( 1,5-Dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-oxalylamino<oxalylamino>)-[1, 1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine-2-carboxylic acid formate (10mg, 14.67μmol) and DIPEA (6.64mg, 51.35 μmol) in THF (2 mL), reacted at this temperature for 10 minutes, then added NaBH 4 (0.83 mg, 22.01 μmol) in water (0.5 mL), slowly raised to room temperature and stirred for 1 hour. The reaction solution was separated and purified by prep-HPLC to obtain the title compound as an off-white solid (2.3 mg, 23%).
MS m/z(ESI):621.2[M+H] +. MS m/z(ESI):621.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.90(s,1H),8.35(dd,J＝8.3,1.6Hz,1H),7.75(d,J＝7.5Hz,1H),7.67(dd,J＝7.7,1.8Hz,1H),7.54(t,J＝7.6Hz,1H),7.48 (t,J＝7.9Hz,1H),7.40(dd,J＝7.5,1.7Hz,1H),7.25(d,J＝7.5Hz,1H),7.18(d,J＝7.5Hz,1H),3.91(s,3H),3.90(s,3H),3.79-3.74(m,1H),3.53-3.47(m,1H),3.41-3.30(s,7H),2.81-2.75(m,1H),2.72-2.63(s,4H),2.38(s,3H),2.00-1.80(m,2H). 1 H NMR (400MHz, DMSO-d 6 )δ9.90(s, 1H), 8.35(dd, J=8.3, 1.6Hz, 1H), 7.75(d, J=7.5Hz, 1H), 7.67(dd, J=7.7,1.8Hz,1H),7.54(t,J=7.6Hz,1H),7.48(t,J=7.9Hz,1H),7.40(dd,J=7.5,1.7Hz,1H),7.25( d,J=7.5Hz,1H),7.18(d,J=7.5Hz,1H),3.91(s,3H),3.90(s,3H),3.79-3.74(m,1H),3.53-3.47(m ,1H),3.41-3.30(s,7H),2.81-2.75(m,1H),2.72-2.63(s,4H),2.38(s,3H),2.00-1.80(m,2H).
实施例13Example 13
N-(2,2'-二氯-3'-(5-((3-羟基-3-(羟甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((3-hydroxy-3-(hydroxymethyl)azetidin-1-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000026
Figure PCTCN2022102195-appb-000026
N-(2,2'-二氯-3'-(5-((3-羟基-3-(羟甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例11。N-(2,2'-dichloro-3'-(5-((3-hydroxy-3-(hydroxymethyl)azetidin-1-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] The preparation of pyridine-2-carboxamide refers to Example 11.
MS m/z(ESI):637.2[M+H] +. MS m/z(ESI):637.2[M+H] + .
实施例14Example 14
N-(3'-(5-((3-乙酰氨基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((3-acetylaminoazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1, 1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000027
Figure PCTCN2022102195-appb-000027
将DIPEA(140.88mg,1.09mmol,189.86μL)加到N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(400mg,726.70μmol)和N-(氮杂环丁烷-3-基)乙酰胺盐酸盐(164.17mg,1.09mmol)的二氯乙烷(5mL)溶液中，室温搅拌1小时，再加入醋酸硼氢化钠(462.18mg,2.18mmol)，室温搅拌2小时。反应液用饱和碳酸氢钠水溶液(10mL)淬灭，用DCM(20mL)提取，有机层减压浓缩，残余物用prep-HPLC分离纯化得标题化合物为白色固体(259mg,51％)。Add DIPEA (140.88 mg, 1.09 mmol, 189.86 μL) to N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (400mg ,726.70μmol) and N-(azetidin-3-yl)acetamide hydrochloride (164.17mg, 1.09mmol) in dichloroethane (5mL) solution, stirred at room temperature for 1 hour, then added boron acetate Sodium hydride (462.18 mg, 2.18 mmol), stirred at room temperature for 2 hours. The reaction solution was quenched with saturated aqueous sodium bicarbonate (10 mL), extracted with DCM (20 mL), the organic layer was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (259 mg, 51%).
MS m/z(ESI):648.2[M+H] +. MS m/z(ESI):648.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.90(s,1H),8.38-8.29(m,2H),7.71(d,J＝ 7.5Hz,1H),7.68(dd,J＝7.7,1.8Hz,1H),7.54(t,J＝7.6Hz,1H),7.48(t,J＝7.9Hz,1H),7.40(dd,J＝7.6,1.7Hz,1H),7.27(d,J＝7.5Hz,1H),7.18(dd,J＝7.6,1.6Hz,1H),4.30(q,J＝6.9Hz,1H),3.91(s,3H),3.90(s,3H),3.62-3.56(m,4H),3.39(s,2H),3.00-2.93(m,2H),2.75-2.64(m,4H),2.40(s,3H),1.80(s,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.90(s, 1H), 8.38-8.29(m, 2H), 7.71(d, J＝7.5Hz, 1H), 7.68(dd, J＝7.7, 1.8 Hz, 1H), 7.54(t, J=7.6Hz, 1H), 7.48(t, J=7.9Hz, 1H), 7.40(dd, J=7.6, 1.7Hz, 1H), 7.27(d, J=7.5 Hz, 1H), 7.18(dd, J=7.6, 1.6Hz, 1H), 4.30(q, J=6.9Hz, 1H), 3.91(s, 3H), 3.90(s, 3H), 3.62-3.56(m ,4H),3.39(s,2H),3.00-2.93(m,2H),2.75-2.64(m,4H),2.40(s,3H),1.80(s,3H).
实施例15Example 15
N-(3'-(5-((3-(乙酰氨基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((3-(acetylaminomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 -Formamide
Figure PCTCN2022102195-appb-000028
Figure PCTCN2022102195-appb-000028
第一步：叔-丁基((1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-基)甲基)氨基甲酸酯的制备The first step: tert-butyl ((1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H- Imidazo[4,5-c]pyridine-2-carbobalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridine-3 Preparation of -yl) methyl) azetidin-3-yl) methyl) carbamate
Figure PCTCN2022102195-appb-000029
Figure PCTCN2022102195-appb-000029
将DIPEA(35.22mg,272.51μmol,47.47μL)加到N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(100mg,181.67μmol)和3-BOC-氨甲基氮杂环丁烷盐酸盐(60.69mg,272.51μmol)的二氯乙烷(2mL)溶液中，室温搅拌0.5小时，再加入醋酸硼氢化钠(191.67mg,908.37μmol)，室温搅拌16小时。反应液用DCM(20mL)稀释，然后用饱和碳酸氢钠水溶液(10mL)洗涤，有机层减压浓缩，得标题化合物为棕色油状物(75mg,58％)。Add DIPEA (35.22 mg, 272.51 μmol, 47.47 μL) to N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (100mg , 181.67μmol) and 3-BOC-aminomethylazetidine hydrochloride (60.69mg, 272.51μmol) in dichloroethane (2mL) solution, stirred at room temperature for 0.5 hours, then added sodium acetate borohydride ( 191.67mg, 908.37μmol), stirred at room temperature for 16 hours. The reaction solution was diluted with DCM (20 mL), washed with saturated aqueous sodium bicarbonate (10 mL), and the organic layer was concentrated under reduced pressure to obtain the title compound as a brown oil (75 mg, 58%).
MS m/z(ESI):720.2[M+H] +. MS m/z(ESI):720.2[M+H] + .
第二步：N-(3'-(5-((3-(氨基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备The second step: N-(3'-(5-((3-(aminomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2' -Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Preparation of pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000030
Figure PCTCN2022102195-appb-000030
室温下将TFA(0.3mL)加到叔-丁基((1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-基)甲基)氨基甲酸酯(75mg,104.07μmol) 的DCM(0.7mL)溶液中，室温搅拌2小时。反应液减压浓缩，残余物用DCM(10mL)稀释，再减压浓缩，残余物在碳酸氢钠水溶液(5mL)和DCM(10mL)中分液，有机层用无水硫酸钠干燥，过滤，减压浓缩，得标题化合物为棕色油状物(60mg,93％)。TFA (0.3 mL) was added to tert-butyl ((1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7 -Tetrahydro-1H-imidazo[4,5-c]pyridine-2-oxamoylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2- A solution of methoxypyridin-3-yl)methyl)azetidin-3-yl)methyl)carbamate (75 mg, 104.07 μmol) in DCM (0.7 mL) was stirred at room temperature for 2 hours. The reaction solution was concentrated under reduced pressure, the residue was diluted with DCM (10 mL), and then concentrated under reduced pressure. The residue was partitioned between aqueous sodium bicarbonate (5 mL) and DCM (10 mL), and the organic layer was dried over anhydrous sodium sulfate and filtered. Concentration under reduced pressure gave the title compound as a brown oil (60 mg, 93%).
MS m/z(ESI):620.2[M+H] +. MS m/z(ESI):620.2[M+H] + .
第三步：N-(3'-(5-((3-(乙酰氨基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备The third step: N-(3'-(5-((3-(acetylaminomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ] Preparation of pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000031
Figure PCTCN2022102195-appb-000031
-20℃下将乙酰氯(11.38mg,145.03μmol,10.35μL)的DCM(0.1mL)溶液滴加到N-(3'-(5-((3-(氨基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(60mg,96.69μmol)、DIPEA(37.49mg,290.06μmol,50.52μL)的DCM(1mL)溶液中，升至室温反应10分钟。反应液减压浓缩，残余物用prep-HPLC分离纯化得标题化合物为白色固体(8.1mg,11％)。A solution of acetyl chloride (11.38mg, 145.03μmol, 10.35μL) in DCM (0.1mL) was added dropwise to N-(3'-(5-((3-(aminomethyl)azetidine-1 -yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl -4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (60 mg, 96.69 μmol), DIPEA (37.49 mg, 290.06 μmol, 50.52 μL) in DCM ( 1 mL) solution, raised to room temperature for 10 minutes. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (8.1 mg, 11%).
MS m/z(ESI):662.2[M+H] +. MS m/z(ESI):662.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.90(s,1H),8.35(d,J＝8.2Hz,1H),7.91(s,1H),7.72-7.64(m,2H),7.54(t,J＝7.6Hz,1H),7.48(t,J＝7.9Hz,1H),7.40(dd,J＝7.6,1.7Hz,1H),7.26(d,J＝7.5Hz,1H),7.18(d,J＝7.6Hz,1H),3.91(s,3H),3.90(s,3H),3.54(s,2H),3.37(s,2H),3.29(s,2H),3.24(d,J＝6.4Hz,2H),2.94(t,J＝6.4Hz,2H),2.73-2.64(q,4H),2.38(s,3H),2.35-2.30(m,1H),1.79(s,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.90(s, 1H), 8.35(d, J=8.2Hz, 1H), 7.91(s, 1H), 7.72-7.64(m, 2H), 7.54( t,J=7.6Hz,1H),7.48(t,J=7.9Hz,1H),7.40(dd,J=7.6,1.7Hz,1H),7.26(d,J=7.5Hz,1H),7.18( d,J=7.6Hz,1H),3.91(s,3H),3.90(s,3H),3.54(s,2H),3.37(s,2H),3.29(s,2H),3.24(d,J =6.4Hz, 2H), 2.94(t, J=6.4Hz, 2H), 2.73-2.64(q, 4H), 2.38(s, 3H), 2.35-2.30(m, 1H), 1.79(s, 3H) .
实施例16Example 16
N-(2,2'-二氯-3'-(5-((3-(氰基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((3-(cyanomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl) -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 -Formamide
Figure PCTCN2022102195-appb-000032
Figure PCTCN2022102195-appb-000032
将DIPEA(9.39mg,72.67μmol,12.66μL)加到N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡 啶-2-甲酰胺(20mg,36.33μmol)和2-(氮杂环丁烷-3-基)乙腈盐酸盐(9.64mg,72.67μmol)的二氯乙烷(0.5mL)溶液中，室温搅拌0.5小时，再加入醋酸硼氢化钠(38.33mg,181.67μmol)，室温搅拌1小时。反应液用DCM(10mL)稀释，然后用饱和碳酸氢钠水溶液(5mL)洗涤，有机层减压浓缩，残余物用prep-HPLC分离纯化得标题化合物为白色固体(18.6mg,76％)。Add DIPEA (9.39 mg, 72.67 μmol, 12.66 μL) to N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (20mg ,36.33μmol) and 2-(azetidin-3-yl)acetonitrile hydrochloride (9.64mg, 72.67μmol) in dichloroethane (0.5mL) solution, stirred at room temperature for 0.5 hours, then added boron acetate Sodium hydride (38.33 mg, 181.67 μmol), stirred at room temperature for 1 hour. The reaction solution was diluted with DCM (10 mL), washed with saturated aqueous sodium bicarbonate (5 mL), the organic layer was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (18.6 mg, 76%).
MS m/z(ESI):630.2[M+H] +. MS m/z(ESI):630.2[M+H] + .
实施例17Example 17
N-(2,2'-二氯-3'-(5-((3-(二甲基氨基甲酰)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((3-(dimethylcarbamoyl)azetidin-1-yl)methyl)-6-methoxypyridine-2- base)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-Carboxamide
Figure PCTCN2022102195-appb-000033
Figure PCTCN2022102195-appb-000033
将DIPEA(28.18mg,218.01μmol,37.97μL)加到N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(60mg,109.00μmol)和N,N-二甲基氮杂环丁烷-3-甲酰胺盐酸盐(27.94mg,218.01μmol)的二氯乙烷(2mL)溶液中，室温搅拌0.5小时，再加入醋酸硼氢化钠(34.50mg,163.51μmol)，室温搅拌0.5小时。反应液用DCM(10mL)稀释，然后用饱和碳酸氢钠水溶液(5mL)洗涤，有机层减压浓缩，残余物用prep-HPLC分离纯化得标题化合物为白色固体(28.3mg,36％)。Add DIPEA (28.18 mg, 218.01 μmol, 37.97 μL) to N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (60mg , 109.00μmol) and N,N-dimethylazetidine-3-carboxamide hydrochloride (27.94mg, 218.01μmol) in dichloroethane (2mL) solution, stirred at room temperature for 0.5 hours, then added Sodium acetate borohydride (34.50mg, 163.51μmol), stirred at room temperature for 0.5 hours. The reaction solution was diluted with DCM (10 mL), washed with saturated aqueous sodium bicarbonate (5 mL), the organic layer was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (28.3 mg, 36%).
MS m/z(ESI):662.2[M+H] +. MS m/z(ESI):662.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.83(s,1H),8.28(dd,J＝8.2,1.5Hz,1H),7.66-7.57(m,2H),7.47(t,J＝7.6Hz,1H),7.41(t,J＝8.0Hz,1H),7.33(dd,J＝7.6,1.7Hz,1H),7.19(d,J＝7.4Hz,1H),7.11(dd,J＝7.6,1.5Hz,1H),3.84(s,3H),3.83(s,3H),3.47(s,2H),3.47-3.41(m,3H),3.31(s,2H),3.16(t,J＝3.6Hz,2H),2.76(s,3H),2.74(s,3H),2.67-2.58(m,4H),2.32(s,3H). 1 H NMR (400MHz, DMSO-d 6 )δ9.83(s, 1H), 8.28(dd, J=8.2, 1.5Hz, 1H), 7.66-7.57(m, 2H), 7.47(t, J=7.6 Hz, 1H), 7.41(t, J=8.0Hz, 1H), 7.33(dd, J=7.6, 1.7Hz, 1H), 7.19(d, J=7.4Hz, 1H), 7.11(dd, J=7.6 ,1.5Hz,1H),3.84(s,3H),3.83(s,3H),3.47(s,2H),3.47-3.41(m,3H),3.31(s,2H),3.16(t,J= 3.6Hz,2H),2.76(s,3H),2.74(s,3H),2.67-2.58(m,4H),2.32(s,3H).
实施例18Example 18
N-(2,2'-二氯-3'-(6-甲氧基-5-((3-(甲基氨基甲酰)吖丁啶-1-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(6-methoxy-5-((3-(methylcarbamoyl)azetidin-1-yl)methyl)pyridin-2-yl )-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine- 2-formamide
Figure PCTCN2022102195-appb-000034
Figure PCTCN2022102195-appb-000034
N-(2,2'-二氯-3'-(6-甲氧基-5-((3-(甲基氨基甲酰)吖丁啶-1-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例11。N-(2,2'-dichloro-3'-(6-methoxy-5-((3-(methylcarbamoyl)azetidin-1-yl)methyl)pyridin-2-yl )-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine- The preparation of 2-formamide refers to Example 11.
MS m/z(ESI):648.2[M+H] +. MS m/z(ESI):648.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.90(s,1H),8.35(dd,J＝8.3,1.5Hz,1H),7.80-7.74(m,1H),7.73-7.64(m,2H),7.54(t,J＝7.6Hz,1H),7.48(t,J＝7.9Hz,1H),7.40(dd,J＝7.6,1.7Hz,1H),7.26(d,J＝7.5Hz,1H),7.18(dd,J＝7.6,1.6Hz,1H),3.91(s,3H),3.90(s,3H),3.54(s,2H),3.46-3.40(m,2H),3.36(s,2H),3.24-3.17(m,2H),3.18-3.10(m,1H),2.74-2.66(m,4H),2.58(d,J＝4.6Hz,3H),2.38(s,3H). 1 H NMR (400MHz,DMSO-d 6 )δ9.90(s,1H),8.35(dd,J＝8.3,1.5Hz,1H),7.80-7.74(m,1H),7.73-7.64(m,2H ),7.54(t,J=7.6Hz,1H),7.48(t,J=7.9Hz,1H),7.40(dd,J=7.6,1.7Hz,1H),7.26(d,J=7.5Hz,1H ),7.18(dd,J＝7.6,1.6Hz,1H),3.91(s,3H),3.90(s,3H),3.54(s,2H),3.46-3.40(m,2H),3.36(s, 2H),3.24-3.17(m,2H),3.18-3.10(m,1H),2.74-2.66(m,4H),2.58(d,J＝4.6Hz,3H),2.38(s,3H).
实施例19Example 19
N-(2,2'-二氯-3'-(5-((6-羟基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((6-hydroxy-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000035
Figure PCTCN2022102195-appb-000035
N-(2,2'-二氯-3'-(5-((6-羟基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例11。N-(2,2'-dichloro-3'-(5-((6-hydroxy-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] The preparation of pyridine-2-carboxamide refers to Example 11.
MS m/z(ESI):647.2[M+H] +. MS m/z(ESI):647.2[M+H] + .
实施例20Example 20
N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methanol Oxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4 ,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000036
Figure PCTCN2022102195-appb-000036
第一步：叔-丁基6-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-羧酸酯的制备The first step: tert-butyl 6-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl ) methyl)-2,6-diazaspiro [3.3] the preparation of heptane-2-carboxylate
Figure PCTCN2022102195-appb-000037
Figure PCTCN2022102195-appb-000037
将N-(2,2'-二氯-3'-(5-甲酰基-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(195mg,354μmol)和2,6-二氮杂螺[3.3]庚烷-2-甲酸叔丁酯(117mg,590μmol)的二氯乙烷(2mL)溶液于室温搅拌0.5小时，再加入醋酸硼氢化钠(226mg,1.06mmol)，室温搅拌0.5小时。反应液用DCM(20mL)稀释，然后用饱和碳酸氢钠水溶液(10mL)洗涤，有机层减压浓缩，硅胶柱层析分离纯化，得标题化合物为棕色油状物(125mg,48％)。N-(2,2'-dichloro-3'-(5-formyl-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1 , 5-Dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (195 mg, 354 μmol) and 2,6-diazaspiro[ 3.3] A solution of tert-butyl heptane-2-carboxylate (117 mg, 590 μmol) in dichloroethane (2 mL) was stirred at room temperature for 0.5 hours, then sodium acetate borohydride (226 mg, 1.06 mmol) was added, and stirred at room temperature for 0.5 hours. The reaction solution was diluted with DCM (20 mL), washed with saturated aqueous sodium bicarbonate (10 mL), the organic layer was concentrated under reduced pressure, and purified by silica gel column chromatography to obtain the title compound as a brown oil (125 mg, 48%).
MS m/z(ESI):732.2[M+H] +. MS m/z(ESI):732.2[M+H] + .
第二步：N-(3'-(5-(2,6-二氮杂螺[3.3]庚烷-2-基甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备The second step: N-(3'-(5-(2,6-diazaspiro[3.3]heptane-2-ylmethyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ] Preparation of pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000038
Figure PCTCN2022102195-appb-000038
室温下将TFA(0.5mL)加到叔-丁基6-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)-2,6-二氮杂螺[3.3]庚烷-2-羧酸酯(157mg,214μmol)的DCM(2mL)溶液中，室温搅拌1小时。反应液减压浓缩，残余物用DCM(10mL)稀释，再减压浓缩，残余物在碳酸氢钠水溶液(5mL)和DCM(10mL)中分液，有机层用无水硫酸钠干燥，过滤，减压浓缩，得标题化合物为棕色油状物(116mg,86％)。TFA (0.5 mL) was added to tert-butyl 6-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetra Hydrogen-1H-imidazo[4,5-c]pyridine-2-carboxalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxy Pyridin-3-yl)methyl)-2,6-diazaspiro[3.3]heptane-2-carboxylate (157 mg, 214 μmol) in DCM (2 mL) was stirred at room temperature for 1 hour. The reaction solution was concentrated under reduced pressure, the residue was diluted with DCM (10 mL), and then concentrated under reduced pressure. The residue was partitioned between aqueous sodium bicarbonate (5 mL) and DCM (10 mL), and the organic layer was dried over anhydrous sodium sulfate and filtered. Concentration under reduced pressure gave the title compound as a brown oil (116 mg, 86%).
MS m/z(ESI):632.2[M+H] +. MS m/z(ESI):632.2[M+H] + .
第三步：N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备The third step: N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.3]heptane-2-yl)methyl) -6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H- Preparation of imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000039
Figure PCTCN2022102195-appb-000039
0℃下将异丁酰氯(8mg,75.9μmol)的DCM(0.1mL)溶液滴加到N-(3'-(5-(2,6-二氮杂螺[3.3]庚烷-2-基甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(40mg,63.3 μmol)、DIPEA(24mg,190μmol)的DCM(1mL)溶液中，0℃反应1小时。反应液减压浓缩，残余物用prep-HPLC分离纯化得标题化合物为白色固体(7.8mg,17％)。A solution of isobutyryl chloride (8mg, 75.9μmol) in DCM (0.1mL) was added dropwise to N-(3'-(5-(2,6-diazaspiro[3.3]heptane-2-yl) at 0°C Methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4, 5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (40mg, 63.3 μmol), DIPEA (24mg, 190μmol) in DCM (1mL) solution, react at 0°C 1 hour. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (7.8 mg, 17%).
MS m/z(ESI):702.2[M+H] +. MS m/z(ESI):702.2[M+H] + .
实施例21Example 21
N-(2,2'-二氯-3'-(5-((6-(环丙羰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((6-(cyclopropylcarbonyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6 -Methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000040
Figure PCTCN2022102195-appb-000040
N-(2,2'-二氯-3'-(5-((6-(环丙羰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例20。N-(2,2'-dichloro-3'-(5-((6-(cyclopropylcarbonyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6 -Methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo The preparation of [4,5-c]pyridine-2-carboxamide refers to Example 20.
MS m/z(ESI):700.2[M+H] +. MS m/z(ESI):700.2[M+H] + .
实施例22Example 22
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-丙酰-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(6-methoxy-5-((6-propionyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000041
Figure PCTCN2022102195-appb-000041
0℃下将丙酰氯(5.48mg,59.28μmol,5.17μL)的DCM(0.1mL)溶液滴加到N-(3'-(5-(2,6-二氮杂螺[3.3]庚烷-2-基甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(25mg,39.52μmol)、DIPEA(25.54mg,197.60μmol,34.42μL)的DCM(1mL)溶液中，室温反应0.5小时。反应液减压浓缩，残余物用prep-HPLC分离纯化得标题化合物为白色固体(8.9mg,31％)。A solution of propionyl chloride (5.48mg, 59.28μmol, 5.17μL) in DCM (0.1mL) was added dropwise to N-(3'-(5-(2,6-diazaspiro[3.3]heptane- 2-ylmethyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl -4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide (25 mg, 39.52 μmol), DIPEA (25.54 mg, 197.60 μmol, 34.42 μL) in DCM ( 1 mL) solution at room temperature for 0.5 hours. The reaction solution was concentrated under reduced pressure, and the residue was separated and purified by prep-HPLC to obtain the title compound as a white solid (8.9 mg, 31%).
MS m/z(ESI):688.2[M+H] +. MS m/z(ESI):688.2[M+H] + .
实施例23Example 23
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(2,2,2-三氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(2,2,2-trifluoroacetyl)-2,6-diazaspiro[3.3 ]heptane-2-yl)methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7- Tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000042
Figure PCTCN2022102195-appb-000042
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(2,2,2-三氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例20。N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(2,2,2-trifluoroacetyl)-2,6-diazaspiro[3.3 ]heptane-2-yl)methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7- Refer to Example 20 for the preparation of tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide.
MS m/z(ESI):728.2[M+H] +. MS m/z(ESI):728.2[M+H] + .
实施例24Example 24
N-(2,2'-二氯-3'-(5-((6-(2,2-二氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((6-(2,2-difluoroacetyl)-2,6-diazaspiro[3.3]heptan-2-yl) Methyl)-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro -1H-imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000043
Figure PCTCN2022102195-appb-000043
N-(2,2'-二氯-3'-(5-((6-(2,2-二氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例20。N-(2,2'-dichloro-3'-(5-((6-(2,2-difluoroacetyl)-2,6-diazaspiro[3.3]heptan-2-yl) Methyl)-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro - Refer to Example 20 for the preparation of 1H-imidazo[4,5-c]pyridine-2-carboxamide.
MS m/z(ESI):710.2[M+H] +. MS m/z(ESI):710.2[M+H] + .
实施例25Example 25
N-(2,2'-二氯-3'-(5-((6-(2-氰基乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((6-(2-cyanoacetyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H -imidazo[4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000044
Figure PCTCN2022102195-appb-000044
N-(2,2'-二氯-3'-(5-((6-(2-氰基乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例20。N-(2,2'-dichloro-3'-(5-((6-(2-cyanoacetyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H - Refer to Example 20 for the preparation of imidazo[4,5-c]pyridine-2-carboxamide.
MS m/z(ESI):699.2[M+H] +. MS m/z(ESI):699.2[M+H] + .
实施例26Example 26
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(甲磺酰)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(methylsulfonyl)-2,6-diazaspiro[3.3]heptane-2-yl )methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000045
Figure PCTCN2022102195-appb-000045
N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(甲磺酰)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例20。N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(methylsulfonyl)-2,6-diazaspiro[3.3]heptane-2-yl )methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo The preparation of [4,5-c]pyridine-2-carboxamide refers to Example 20.
MS m/z(ESI):710.2[M+H] +. MS m/z(ESI):710.2[M+H] + .
实施例27Example 27
N-(2,2'-二氯-3'-(5-((6-甲酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((6-formyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-6-methoxy Basepyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000046
Figure PCTCN2022102195-appb-000046
N-(2,2'-二氯-3'-(5-((6-甲酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例20。N-(2,2'-dichloro-3'-(5-((6-formyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-6-methoxy Basepyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] The preparation of pyridine-2-carboxamide refers to Example 20.
MS m/z(ESI):660.2[M+H] +. MS m/z(ESI):660.2[M+H] + .
实施例28Example 28
N-(3'-(5-((7-乙酰基-2,7-二氮杂螺[3.5]壬烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((7-acetyl-2,7-diazaspiro[3.5]nonan-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000047
Figure PCTCN2022102195-appb-000047
N-(3'-(5-((7-乙酰基-2,7-二氮杂螺[3.5]壬烷-2-基)甲基)-6-甲氧基吡啶-2-N-(3'-(5-((7-acetyl-2,7-diazaspiro[3.5]nonan-2-yl)methyl)-6-methoxypyridine-2-
基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例1。base)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo The preparation of [4,5-c]pyridine-2-carboxamide refers to Example 1.
MS m/z(ESI):702.2[M+H]+.MS m/z(ESI):702.2[M+H]+.
实施例29Example 29
N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000048
Figure PCTCN2022102195-appb-000048
N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例1。N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] The preparation of pyridine-2-carboxamide refers to Example 1.
MS m/z(ESI):688.2[M+H] +. MS m/z(ESI):688.2[M+H] + .
实施例30Example 30
N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methanol Oxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4 ,5-c]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000049
Figure PCTCN2022102195-appb-000049
N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例1。N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methanol Oxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4 , 5-c] The preparation of pyridine-2-carboxamide refers to Example 1.
MS m/z(ESI):716.2[M+H] +. MS m/z(ESI):716.2[M+H] + .
实施例31Example 31
N-(3'-(5-((6-乙酰氨基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((6-Acetamido-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]pyridine-2-carboxamide
Figure PCTCN2022102195-appb-000050
Figure PCTCN2022102195-appb-000050
N-(3'-(5-((6-乙酰氨基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的制备参照实施例1。N-(3'-(5-((6-Acetamido-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ] The preparation of pyridine-2-carboxamide refers to Example 1.
MS m/z(ESI):688.2[M+H] +. MS m/z(ESI):688.2[M+H] + .
1H NMR(400MHz,DMSO-d 6)δ9.90(s,1H),8.35(dd,J＝8.3,1.6Hz,1H),8.03(d,J＝7.5Hz,1H),7.72-7.65(m,2H),7.54(t,J＝7.7Hz,1H),7.48(t,J＝8.0Hz,1H),7.40(dd,J＝7.6,1.7Hz,1H),7.25(d,J＝7.5Hz,1H),7.18(dd,J＝7.6,1.6Hz,1H),4.08-3.97(m,1H),3.93-3.88(m,6H),3.52(s,2H),3.37(s,2H),3.27(s,2H),3.16(s,2H),2.71-2.64(m,4H),2.43-2.32(m,5H),2.01-1.94(m,2H),1.74(s,3H). 1 H NMR (400MHz, DMSO-d 6 ) δ9.90(s, 1H), 8.35(dd, J=8.3, 1.6Hz, 1H), 8.03(d, J=7.5Hz, 1H), 7.72-7.65( m,2H),7.54(t,J=7.7Hz,1H),7.48(t,J=8.0Hz,1H),7.40(dd,J=7.6,1.7Hz,1H),7.25(d,J=7.5 Hz,1H),7.18(dd,J=7.6,1.6Hz,1H),4.08-3.97(m,1H),3.93-3.88(m,6H),3.52(s,2H),3.37(s,2H) ,3.27(s,2H),3.16(s,2H),2.71-2.64(m,4H),2.43-2.32(m,5H),2.01-1.94(m,2H),1.74(s,3H).
生物学测试评价Biology Test Evaluation
以下结合测试例进一步描述解释本发明，但这些实施例并非意味着限制本发明的范围。The following further describes and explains the present invention in combination with test examples, but these examples are not meant to limit the scope of the present invention.
测试例1本发明化合物对人PD-1/PD-L1结合抑制作用的测定Test Example 1 Determination of the binding inhibitory effect of the compound of the present invention on human PD-1/PD-L1
实验目的：该测试例的目的是测量化合物对人PD-1/PD-L1结合抑制的活性。Purpose of the experiment: The purpose of this test case is to measure the activity of the compound on human PD-1/PD-L1 binding inhibition.
实验仪器：离心机(5810R)购自Eppendorf公司，移液器购自Eppendorf或Rainin公司，酶标仪购自美国BioTek公司，型号为H1MFD全功能酶标仪。Experimental equipment: a centrifuge (5810R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, a microplate reader was purchased from BioTek, USA, and the model was H1MFD full-function microplate reader.
实验试剂：Binding Domain diluent buffer购自Cisbio公司，货号为62DLBDDF；MAb Anti-6HIS Eu cryptate Gold购自Cisbio公司，货号为61HI2KLA；PAb Anti Human IgG-XL665购自Cisbio公司，货号为61HFCXLB；PD-L1-His蛋白购自Abcam公司，货号为ab167713；PD-1-Fc蛋白购自R&D公司，货号为1086-PD；Detection buffer购自Cisbio公司，货号为62SDBRDD；384孔板购自PerkinElmer公司，货号为6007290。Experimental reagents: Binding Domain diluent buffer was purchased from Cisbio Company, the article number was 62DLBDDF; MAb Anti-6HIS Eu cryptate Gold was purchased from Cisbio Company, the article number was 61HI2KLA; PAb Anti Human IgG-XL665 was purchased from Cisbio Company, the article number was 61HFCXLB; PD-L1 -His protein was purchased from Abcam Company, the article number is ab167713; PD-1-Fc protein was purchased from R&D Company, the article number was 1086-PD; Detection buffer was purchased from Cisbio Company, the article number was 62SDBRDD; 384-well plates were purchased from PerkinElmer Company, the article number was 6007290.
实验方法：为了测试化合物对人PD-1/PD-L1结合的抑制活性，本实验采用时间分辨荧光共振能量转移(TR-FRET)的方法测试化合物对人PD-1/PD-L1结合的抑制作用，并得出化合物对人PD-1/PD-L1结合抑制活性的半数抑制浓度IC 50。 Experimental method: In order to test the inhibitory activity of the compound on the binding of human PD-1/PD-L1, the method of time-resolved fluorescence resonance energy transfer (TR-FRET) was used in this experiment to test the inhibition of the compound on the binding of human PD-1/PD-L1 The effect, and the half-inhibitory concentration IC 50 of the compound's inhibitory activity on human PD-1/PD-L1 binding was obtained.
具体实验操作如下：The specific experimental operation is as follows:
本实验在384孔板中进行，总反应体系为20μL，将化合物使用实验缓冲液Binding Domain diluent buffer(Cisbio#62DLBDDF)稀释成不同的浓度(10μM或1μM起始，3倍稀释，11个剂量)，将PD-L1-His蛋白(19～238氨基酸)(Abcam#ab167713)用实验缓冲液稀释为10nM，将PD-1-Fc蛋白(25～167氨基酸)(R&D#1086-PD)用实验缓冲液稀释为20nM，将化合物、PD-L1蛋白以及PD-1蛋白分别加入到384孔板中，总体积为10μL，放入离心机1000rpm离心1分钟混合均匀，室温孵育反应60分钟，每孔加入10μL使用HTRF Detection buffer稀释的MAb Anti-6HIS Eu cryptate Gold(Cisbio#61HI2KLA)以及PAb Anti Human IgG-XL665(Cisbio#61HFCXLB)混合溶液，放入离心机1000rpm离心1分钟混合均匀，室温反应3小时或者4℃反应过夜，放入酶标仪(BioTek Synergy H1)进行读数，激发波长为340nm，发射波长为620nm及665nm。This experiment was carried out in a 384-well plate, the total reaction system was 20 μL, and the compound was diluted into different concentrations (10 μM or 1 μM starting, 3-fold dilution, 11 doses) using the experimental buffer Binding Domain diluent buffer (Cisbio#62DLBDDF) , Dilute PD-L1-His protein (19-238 amino acids) (Abcam#ab167713) to 10nM with experimental buffer, and dilute PD-1-Fc protein (25-167 amino acids) (R&D#1086-PD) with experimental buffer The solution was diluted to 20nM, and the compound, PD-L1 protein, and PD-1 protein were added to a 384-well plate, with a total volume of 10 μL, placed in a centrifuge at 1000rpm for 1 minute to mix well, incubated at room temperature for 60 minutes, and added to each well. 10 μL mixed solution of MAb Anti-6HIS Eu cryptate Gold (Cisbio#61HI2KLA) and PAb Anti Human IgG-XL665 (Cisbio#61HFCXLB) diluted with HTRF Detection buffer, placed in a centrifuge at 1000rpm for 1 minute to mix well, and reacted at room temperature for 3 hours or React overnight at 4°C, put into a microplate reader (BioTek Synergy H1) for reading, the excitation wavelength is 340nm, and the emission wavelength is 620nm and 665nm.
实验数据处理方法：Experimental data processing method:
计算620nm及665nm下荧光读数的比值(665nm/620nm)，并计算抑制率，将化合物浓度以及抑制率使用Graphpad Prism软件进行非线性回归拟合，得出IC 50值，如下表9所示。 The ratio of the fluorescence readings at 620nm and 665nm (665nm/620nm) was calculated, and the inhibition rate was calculated. The compound concentration and inhibition rate were used for nonlinear regression fitting using Graphpad Prism software to obtain the IC 50 value, as shown in Table 9 below.
表9化合物对人PD-1/PD-L1结合的IC 50值 Table 9 IC 50 values of compounds binding to human PD-1/PD-L1
Figure PCTCN2022102195-appb-000051
Figure PCTCN2022102195-appb-000051
实验结论：本发明的实施例化合物在PD-1/PD-L1抑制试验中，显示出良好结合抑制活性。Experimental conclusion: the compounds of the examples of the present invention showed good binding inhibitory activity in the PD-1/PD-L1 inhibition test.
测试例2本发明化合物诱导肿瘤细胞表面PD-L1蛋白内吞活性的测定Test example 2 Determination of endocytic activity of PD-L1 protein on tumor cell surface induced by compounds of the present invention
实验目的：该测试例的目的是测试化合物诱导肿瘤细胞表面PD-L1内吞的活性。Purpose of the experiment: The purpose of this test case is to test the activity of the compound in inducing endocytosis of PD-L1 on the surface of tumor cells.
实验仪器：离心机(5702R)购自Eppendorf公司，移液器购自Eppendorf或Rainin公司，流式细胞仪购自Beckman Coulter，型号为DxFlex。Experimental equipment: a centrifuge (5702R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, and a flow cytometer was purchased from Beckman Coulter, model DxFlex.
实验试剂：PE Mouse Anti-Human CD274抗体购自BD Pharmingen公司，货号为557924；BSA购自Sigma公司，货号为B2064-100G；PBS购自Gibco公司，货号为10010049；24孔板购自Corning公司，货号为3526。Experimental reagents: PE Mouse Anti-Human CD274 antibody was purchased from BD Pharmingen Company, the article number is 557924; BSA was purchased from Sigma Company, the article number was B2064-100G; PBS was purchased from Gibco Company, the article number was 10010049; 24-well plates were purchased from Corning Company, The item number is 3526.
实验方法：为了测试化合物诱导PD-L1内吞的活性，本实验通过用荧光标记的抗PD-L1抗体检测肿瘤细胞表面PD-L1的变化程度来测试化合物诱导PD-L1内吞的活性，并得出化合物诱导PD-L1蛋白内吞活性的IC 50。 Experimental method: In order to test the activity of compounds inducing PD-L1 endocytosis, this experiment tested the activity of compounds inducing PD-L1 endocytosis by detecting the degree of change of PD-L1 on the surface of tumor cells with fluorescently labeled anti-PD-L1 antibodies, and The IC 50 of the compounds inducing the endocytic activity of PD-L1 protein was obtained.
具体实验操作如下：The specific experimental operation is as follows:
收集高表达hPD-L1的小鼠结肠癌细胞MC38-hPDL1，调节成合适的密度铺于24孔板，放入37℃，5％CO 2培养箱贴壁过夜。将化合物使用培养基配制成不同的浓度后加入到24孔板中，并设置溶媒对照孔，放入37℃，5％CO 2培养箱中孵育16小时后取出24孔板，收集板内不同处理的MC38-hPDL1细胞，使用FACS buffer(含0.5％BSA的PBS)洗涤一遍后，用FACS buffer将细胞配制成合适的密度，加入PE Mouse Anti-Human CD274抗体(BD Pharmingen#557924)，室温摇床避光孵育30分钟，将细胞使用FACS buffer洗涤两次后，使用100μL PBS重悬，使用流式细胞仪检测细胞表面的荧光信号，设置同型对照为阴性对照。 The mouse colon cancer cells MC38-hPDL1 with high expression of hPD-L1 were collected, adjusted to a suitable density, spread on a 24-well plate, and placed in a 37°C, 5% CO 2 incubator to adhere to the wall overnight. The compound was prepared into different concentrations in the medium and added to the 24-well plate, and the solvent control well was set, and incubated in a 37°C, 5% CO2 incubator for 16 hours, then the 24-well plate was taken out, and different treatments in the plate were collected. MC38-hPDL1 cells were washed once with FACS buffer (PBS containing 0.5% BSA), and the cells were prepared to a suitable density with FACS buffer, and PE Mouse Anti-Human CD274 antibody (BD Pharmingen#557924) was added, and shaken at room temperature Incubate in the dark for 30 minutes, wash the cells twice with FACS buffer, resuspend in 100 μL PBS, use flow cytometry to detect the fluorescent signal on the cell surface, and set the isotype control as a negative control.
实验数据处理方法：Experimental data processing method:
用不同处理组的荧光信号计算化合物的内吞率，并将化合物浓度以及内吞率使用GraphPad Prism软件进行非线性回归拟合，得出IC 50值，如表10所示。 The endocytosis rate of the compound was calculated using the fluorescence signals of different treatment groups, and the concentration of the compound and the endocytosis rate were used for nonlinear regression fitting using GraphPad Prism software to obtain the IC 50 value, as shown in Table 10.
表10化合物在肿瘤细胞表面PD-L1的最大内吞率及IC 50值 Table 10 The maximum endocytosis rate and IC 50 value of the compounds on the surface of tumor cells PD-L1
实施例编号Example number MC38最大内吞率MC38 maximum endocytosis rate MC38IC 50(nM) MC38IC 50 (nM)
11 95％95% 1.21.2
22 95％95% 1.21.2
33 97％97% 0.40.4
44 97％97% 0.40.4
55 96％96% 0.40.4
66 96％96% 0.70.7
77 97％97% 1.01.0
88 95％95% 1.01.0
99 97％97% 1.21.2
1010 98％98% 3.13.1
1111 93％93% NANA
1212 94％94% NANA
1313 97％97% 0.90.9
1414 96％96% 0.60.6
1515 97％97% NANA
1616 97％97% NANA
1717 95％95% NANA
1818 96％96% NANA
1919 97％97% 0.70.7
2020 96％96% 0.70.7
21twenty one 95％95% 1.11.1
22twenty two 97％97% 0.40.4
23twenty three 96％96% NANA
24twenty four 97％97% 0.80.8
2525 97％97% NANA
2626 97％97% 0.30.3
2727 96％96% NANA
2828 96％96% NANA
2929 96％96% NANA
3030 96％96% NANA
3131 97％97% NANA
实验结论：本发明所示的化合物能够很好的诱导肿瘤细胞表面PD-L1的内吞。Experimental conclusion: the compounds shown in the present invention can well induce the endocytosis of PD-L1 on the surface of tumor cells.
测试例3本发明化合物对CHO-PDL1/Jurkat-PD1细胞通路的抑制作用Test example 3 Inhibitory effect of the compound of the present invention on CHO-PDL1/Jurkat-PD1 cell pathway
实验目的：该测试例的目的是测试化合物对CHO-PDL1/Jurkat-PD1细胞通路的抑制作用。Experimental purpose: The purpose of this test case is to test the inhibitory effect of the compound on the CHO-PDL1/Jurkat-PD1 cellular pathway.
实验仪器：离心机(5810R)购自Eppendorf公司，移液器购自Eppendorf或Rainin公司，酶标仪购自美国BioTek公司，型号为H1MFD全功能酶标仪。Experimental equipment: a centrifuge (5810R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, a microplate reader was purchased from BioTek, USA, and the model was H1MFD full-function microplate reader.
实验试剂：CHO-PDL1细胞及NFAT-luc2/PD1 Jurkat细胞来源于Promega公司，货号为J1252；RPMI 1640购自Gibco公司，货号为22400089；FBS购自Gibco公司，货号为10091148；One-Glo试剂购自Promega公司，货号为E6120；96孔板购自Corning公司，货号为3610。Experimental reagents: CHO-PDL1 cells and NFAT-luc2/PD1 Jurkat cells were purchased from Promega Company, product number J1252; RPMI 1640 was purchased from Gibco Company, product number 22400089; FBS was purchased from Gibco Company, product number 10091148; One-Glo reagent was purchased from The 96-well plate was purchased from Promega Company, the product number is E6120; the 96-well plate was purchased from Corning Company, the product number is 3610.
实验方法：本实验中包含两株稳转细胞株，稳转了PD-L1的CHO-K1细胞以及稳转了PD-1以及NFAT荧光素酶报告基因的Jurkat细胞，化合物与两株细 胞共孵育后，由于其对PD-1/PD-L1通路的抑制作用使Jurkat细胞的激活程度发生变化，从而使Jurkat细胞中下游的报告基因NFAT水平发生变化，通过检测荧光素酶的信号值来表征NFAT的水平，以此表征测试化合物对CHO-PDL1/Jurkat-PD1通路的抑制作用。Experimental method: In this experiment, two stably transfected cell lines were included, CHO-K1 cells stably transfected with PD-L1 and Jurkat cells stably transfected with PD-1 and NFAT luciferase reporter gene, and the compound was co-incubated with the two cell lines Finally, due to its inhibitory effect on the PD-1/PD-L1 pathway, the activation degree of Jurkat cells changes, so that the level of the reporter gene NFAT in the downstream of Jurkat cells changes, and the signal value of luciferase is used to characterize NFAT In order to characterize the inhibitory effect of the test compound on the CHO-PDL1/Jurkat-PD1 pathway.
具体实验操作如下：The specific experimental operation is as follows:
培养CHO-PDL1细胞(Promega,#J1252)至合适的细胞密度，消化后使用完全培养基重悬为合适密度的细胞悬液，每孔100μL铺于96孔板(Corning,#3610)，放入37℃，5％CO 2培养箱贴壁培养过夜，使用分析培养基(RPMI 1640+2％FBS,RPMI 1640货号为Gibco,#22400089，FBS货号为Gibco,#10091148)配制不同浓度的化合物溶液，收集NFAT-luc2/PD1 Jurkat细胞(Promega,#J1252)，使用分析培养基重悬为合适的细胞密度。吸去CHO-PDL1细胞培养板的上清，加入40μL配制好的化合物溶液以及40μL重悬的NFAT-luc2/PD1 Jurkat细胞，放入37℃，5％CO 2培养箱孵育6小时，取出细胞板，每孔加入40μL One-Glo试剂(Promega,#E6120)，振荡混匀后室温避光孵育10分钟，放入酶标仪中使用发光程序读取细胞板的发光值。 Cultivate CHO-PDL1 cells (Promega, #J1252) to a suitable cell density, digest and resuspend the cell suspension with complete medium to a suitable density, spread 100 μL per well on a 96-well plate (Corning, #3610), put 37°C, 5% CO 2 incubator adherent culture overnight, use analysis medium (RPMI 1640+2% FBS, RPMI 1640 product number is Gibco, #22400089, FBS product number is Gibco, #10091148) to prepare compound solutions with different concentrations, NFAT-luc2/PD1 Jurkat cells (Promega, #J1252) were collected and resuspended to an appropriate cell density using assay medium. Aspirate the supernatant of the CHO-PDL1 cell culture plate, add 40 μL of the prepared compound solution and 40 μL of resuspended NFAT-luc2/PD1 Jurkat cells, and incubate for 6 hours in a 37°C, 5% CO 2 incubator, then take out the cell plate , add 40 μL of One-Glo reagent (Promega, #E6120) to each well, shake and mix well, incubate at room temperature in the dark for 10 minutes, put it into a microplate reader and use the luminescence program to read the luminescence value of the cell plate.
实验数据处理方法：Experimental data processing method:
根据化合物浓度以及发光信号值拟合EC 50值，并计算每个化合物相对于未加药组信号值的最大诱导倍数，具体如表11所示。 The EC 50 value was fitted according to the compound concentration and the luminescent signal value, and the maximum induction factor of each compound relative to the signal value of the untreated group was calculated, as shown in Table 11.
表11化合物对CHO-PDL1/Jurkat-PD1细胞EC 50及化合物相对于未加药组信号值的最大诱导倍数 Table 11 Compounds on CHO-PDL1/Jurkat-PD1 cells EC 50 and the maximum induction fold of the compound relative to the signal value of the untreated group
Figure PCTCN2022102195-appb-000052
Figure PCTCN2022102195-appb-000052
实验结论：本发明的实施例化合物在对CHO-PDL1/Jurkat-PD1细胞通路的抑制试验中，显示出较好的抑制活性。Experimental conclusion: the compounds of the examples of the present invention showed good inhibitory activity in the inhibition test of CHO-PDL1/Jurkat-PD1 cell pathway.
测试例4发明化合物结合提升PD-L1蛋白稳定性(熔解温度)的测定Test Example 4 Determination of the Combination of Invention Compounds to Improve PD-L1 Protein Stability (Melting Temperature)
实验目的：测试化合物提升PD-L1蛋白稳定性，使蛋白熔解温度上升的能力。The purpose of the experiment: To test the ability of the compound to improve the stability of PD-L1 protein and increase the melting temperature of the protein.
实验仪器：定量PCR仪(Quantstudio6 Flex)购自Life公司，移液器购自Eppendorf或Rainin公司。Experimental equipment: Quantitative PCR instrument (Quantstudio6 Flex) was purchased from Life Company, and the pipette was purchased from Eppendorf or Rainin Company.
实验试剂：Protein Thermal Shift TM Dye Kit购自Thermofisher公司，货号为4461146；PD-L1蛋白购自Acro Biosystems公司，货号为PD1-H5229；HEPES,1M Buffer Solution购自Thermofisher公司，货号为15630080；NaCl购自国药集团化学试剂有限公司，货号为10019318。 Experimental reagents: Protein Thermal Shift TM Dye Kit was purchased from Thermofisher Company, product number 4461146; PD-L1 protein was purchased from Acro Biosystems Company, product number was PD1-H5229; HEPES, 1M Buffer Solution was purchased from Thermofisher Company, product number 15630080; NaCl was purchased from From Sinopharm Chemical Reagent Co., Ltd., the product number is 10019318.
实验方法：本实验通过thermal shift方法测试化合物结合前后PD-L1蛋白熔解温度(Tm)的变化程度来表征化合物提升PD-L1蛋白稳定性的能力从而表现化合物与PD-L1蛋白的结合能力。Experimental method: In this experiment, the thermal shift method was used to test the change degree of the melting temperature (Tm) of the PD-L1 protein before and after the combination of the compound to characterize the ability of the compound to improve the stability of the PD-L1 protein, thereby showing the binding ability of the compound to the PD-L1 protein.
具体实验操作如下：The specific experimental operation is as follows:
配制含10μM HEPES、SYPRO Orange和150mM NaCl的溶液作为实验缓冲液，并加入终浓度为2μM的人PD-L1蛋白。将以上反应混合物分装到8联排PCR管中，每管19.5μL，分别加入0.5μL的测试化合物或者DMSO，则总反应体系为20μL，化合物终浓度为10μM，并设置2.5％DMSO为溶媒对照。将PCR管放入PCR仪中，选取melt curve功能检测不同处理组中PD-L1蛋白的熔解温度(从25℃加热至95℃,0.03℃/s)。Prepare a solution containing 10 μM HEPES, SYPRO Orange and 150 mM NaCl as the experimental buffer, and add human PD-L1 protein at a final concentration of 2 μM. Distribute the above reaction mixture into 8 PCR tubes, 19.5 μL in each tube, add 0.5 μL of the test compound or DMSO respectively, the total reaction system is 20 μL, the final concentration of the compound is 10 μM, and set 2.5% DMSO as the solvent control . Put the PCR tube into the PCR machine, and select the melt curve function to detect the melting temperature of PD-L1 protein in different treatment groups (heating from 25°C to 95°C, 0.03°C/s).
实验数据处理方法：Experimental data processing method:
将PCR仪实验数据文件导入至thermal shift软件，得出每个处理组的熔解温度(Tm)，并减去DMSO溶媒对照组的Tm，得到熔解温度的变化值(ΔTm)，如下表所示：Import the experimental data file of the PCR instrument into the thermal shift software to obtain the melting temperature (Tm) of each treatment group, and subtract the Tm of the DMSO solvent control group to obtain the change value of the melting temperature (ΔTm), as shown in the following table:
表12化合物的熔解温度的变化值The variation value of the melting temperature of the compound of table 12
实施例编号Example number ΔTm(℃)ΔTm(°C)
11 17.917.9
22 15.815.8
33 14.814.8
44 16.516.5
55 16.216.2
66 15.615.6
77 17.117.1
88 13.913.9
99 18.118.1
1010 16.716.7
1111 10.310.3
1212 13.913.9
1313 13.513.5
1414 20.620.6
1515 15.015.0
1616 16.316.3
1717 21.021.0
1818 16.716.7
1919 20.920.9
2020 21.421.4
21twenty one 21.121.1
22twenty two 16.516.5
23twenty three 19.519.5
24twenty four 14.014.0
2525 11.711.7
2626 14.014.0
2727 12.312.3
2828 13.213.2
2929 14.514.5
3030 12.412.4
3131 15.615.6
实验结论：Experimental results:
通过以上方案得出本发明所示的化合物对提升PD-L1蛋白熔解温度的试验中显示提升PD-L1蛋白稳定性的能力。Through the above scheme, it can be concluded that the compounds shown in the present invention have the ability to improve the stability of PD-L1 protein in the test of increasing the melting temperature of PD-L1 protein.
测试例5本发明化合物对NFAT-luc2/PD1 Jurkat细胞以及MC38-hPDL1细胞的体外细胞毒性作用Test example 5 The in vitro cytotoxic effect of the compound of the present invention on NFAT-luc2/PD1 Jurkat cells and MC38-hPDL1 cells
实验目的：测试化合物对NFAT-luc2/PD1 Jurkat细胞以及MC38-hPDL1细胞的体外细胞毒性作用。Experimental purpose: To test the in vitro cytotoxicity of compounds on NFAT-luc2/PD1 Jurkat cells and MC38-hPDL1 cells.
实验仪器：离心机(5810R)购自Eppendorf公司，移液器购自Eppendorf或Rainin公司，酶标仪购自美国BioTek公司，型号为H1MFD全功能酶标仪。Experimental equipment: a centrifuge (5810R) was purchased from Eppendorf, a pipette was purchased from Eppendorf or Rainin, a microplate reader was purchased from BioTek, USA, and the model was H1MFD full-function microplate reader.
实验试剂：RPMI 1640购自Gibco公司，货号为22400089；FBS购自Gibco公司，货号为10091148；PBS购自Gibco公司，货号为10010049；Cell Titer-Glo购自Promega公司，货号为G7573；细胞培养板购自Corning公司，货号为3610。Experimental reagents: RPMI 1640 was purchased from Gibco Company, the product number is 22400089; FBS was purchased from Gibco Company, the product number was 10091148; PBS was purchased from Gibco Company, the product number was 10010049; Cell Titer-Glo was purchased from Promega Company, the product number was G7573; cell culture plate Purchased from Corning Company, Cat. No. 3610.
实验方法：experimental method:
培养细胞至合适的融合度时，收集细胞，使用完全培养基将细胞调整为合适的细胞浓度，将细胞悬液铺于96孔板，每孔90μL，放入37℃，5％CO 2培养箱贴壁过夜，使用DMSO以及培养基配制不同浓度的化合物溶液，设置溶媒对照，将化合物溶液加入到96孔板中，每孔10μL，放入37℃，5％CO 2培养箱中继续培养72h后，加入CellTiter-Glo溶液，振荡混合均匀后，避光孵育10分钟，用BioTek Synergy H1酶标仪进行读数。 When the cells are cultured to a suitable degree of confluence, collect the cells, use the complete medium to adjust the cells to a suitable cell concentration, spread the cell suspension on a 96-well plate, 90 μL per well, and place it in a 37°C, 5% CO 2 incubator Adhere to the wall overnight, use DMSO and medium to prepare compound solutions of different concentrations, set up vehicle control, add the compound solution to a 96-well plate, 10 μL per well, put it in a 37°C, 5% CO 2 incubator and continue to cultivate for 72 hours , add CellTiter-Glo solution, shake and mix well, incubate in the dark for 10 minutes, and read with BioTek Synergy H1 microplate reader.
实验数据处理方法：Experimental data processing method:
使用发光信号值计算抑制率，将浓度以及抑制率使用Graphpad Prism软件进行非线性回归曲线拟合，得到IC 50值，如下表所示： Use the luminescent signal value to calculate the inhibition rate, and use the Graphpad Prism software to perform nonlinear regression curve fitting on the concentration and inhibition rate to obtain the IC 50 value, as shown in the table below:
表13化合物的体外细胞毒性作用In Vitro Cytotoxicity of Table 13 Compounds
Figure PCTCN2022102195-appb-000053
Figure PCTCN2022102195-appb-000053
实验结论：Experimental results:
通过以上结果可知，本发明所示的化合物对NFAT-luc2/PD1 Jurkat以及MC38-hPDL1细胞的毒性作用较弱。From the above results, it can be seen that the compounds shown in the present invention have weak toxic effects on NFAT-luc2/PD1 Jurkat and MC38-hPDL1 cells.
测试例6Balb/C小鼠药代动力学测定Test example 6 Balb/C mouse pharmacokinetics is measured
1.研究目的：1. Research purposes:
以Balb/C小鼠为受试动物，研究以下化合物实施例，在5mg/kg剂量下口服给药在小鼠体内血浆的药代动力学行为。Taking Balb/C mice as the experimental animals, the pharmacokinetics of the following compound examples were orally administered at a dose of 5 mg/kg in mice in vivo.
2.试验方案2. Test plan
2.1试验药品：2.1 Test drugs:
本发明实施例，自制。Embodiment of the present invention, self-made.
2.2试验动物：2.2 Test animals:
Balb/C Mouse 6只/实施例，雄性，上海杰思捷实验动物有限公司，动物生产许可证号(SCXK(沪)2013-0006 N0.311620400001794)。Balb/C Mouse 6/embodiment, male, Shanghai Jiesijie Experimental Animal Co., Ltd., animal production license number (SCXK (Shanghai) 2013-0006 N0.311620400001794).
2.3药物配制：2.3 Drug preparation:
0.5％HPMC(1％Tween 80)，超声溶解，配制为澄清溶液或均一混悬液。0.5% HPMC (1% Tween 80), ultrasonically dissolved, prepared as a clear solution or a homogeneous suspension.
2.3给药：2.3 Administration:
Balb/C小鼠，雄性；禁食一夜后分别p.o.，剂量为5mg/kg，给药体积10mL/kg。Balb/C mice, male; p.o. after fasting overnight, the dose is 5mg/kg, and the administration volume is 10mL/kg.
2.4样品采集：2.4 Sample collection:
小鼠给药前和给药后，在0、0.5、1、2、4、6、8和24小时，采用眼眶采血0.1mL，置于EDTA-K 2试管中，4℃6000rpm离心6min分离血浆，于-80℃保存。 Before and after administration, at 0, 0.5, 1, 2, 4, 6, 8 and 24 hours, 0.1 mL of orbital blood was collected from mice, placed in EDTA-K 2 test tubes, and centrifuged at 6000 rpm at 4°C for 6 minutes to separate plasma , stored at -80°C.
2.5样品处理：2.5 Sample handling:
1)血浆样品40μL加入160μL乙腈沉淀,混合后3500×g离心5～20分钟。1) Add 40 μL of plasma sample to 160 μL of acetonitrile for precipitation, after mixing, centrifuge at 3500×g for 5-20 minutes.
2)取处理后上清溶液100μL进行LC/MS/MS分析待测化合物的浓度。2.6液相分析2) Take 100 μL of the treated supernatant solution and analyze the concentration of the compound to be tested by LC/MS/MS. 2.6 Liquid phase analysis
●液相条件：Shimadzu LC-20AD泵●Liquid phase conditions: Shimadzu LC-20AD pump
●质谱条件:AB Sciex API 4000质谱仪●Mass spectrometer conditions: AB Sciex API 4000 mass spectrometer
●色谱柱：phenomenex Gemiu 5μm C18 50×4.6mm●Column: phenomenonex Gemiu 5μm C18 50×4.6mm
●移动相：A液为0.1％甲酸水溶液，B液为乙腈●Mobile phase: liquid A is 0.1% formic acid aqueous solution, liquid B is acetonitrile
●流速：0.8mL/min●Flow rate: 0.8mL/min
●洗脱时间：0-4.0分钟，洗脱液如下：●Elution time: 0-4.0 minutes, the eluent is as follows:
Figure PCTCN2022102195-appb-000054
Figure PCTCN2022102195-appb-000054
3.试验结果与分析3. Test results and analysis
药代动力学主要参数用WinNonlin 8.2计算得到，小鼠药代实验结果见下表：The main parameters of pharmacokinetics are calculated by WinNonlin 8.2, and the results of pharmacokinetic experiments in mice are shown in the table below:
表14小鼠药代实验结果Table 14 Mouse pharmacokinetic test results
Figure PCTCN2022102195-appb-000055
Figure PCTCN2022102195-appb-000055
4.实验结论：4. Experimental conclusion:
从表中小鼠药代实验结果可以看出，本发明实施例化合物表现出良好的代谢 性质，暴露量AUC和最大血药浓度C max都表现良好。 It can be seen from the results of pharmacokinetic experiments in mice in the table that the compounds of the examples of the present invention exhibit good metabolic properties, and both the exposure AUC and the maximum blood concentration C max are good.
测试例7荷瘤裸鼠血浆和肿瘤的药代动力学测定Test example 7 Pharmacokinetic determination of tumor-bearing nude mouse plasma and tumor
1.研究目的：1. Research purposes:
以荷瘤裸鼠为受试动物，研究以下化合物实施例，在30mg/kg剂量下口服给药在荷瘤裸鼠体内血浆和肿瘤的药代动力学行为。Taking tumor-bearing nude mice as experimental animals, the pharmacokinetic behavior of the following compound examples in plasma and tumor was studied after oral administration at a dose of 30 mg/kg.
2.试验方案2. Test plan
2.1试验药品：2.1 Test drugs:
本发明实施例，自制。Embodiment of the present invention, self-made.
2.2试验动物：2.2 Test animals:
荷瘤裸鼠21只/实施例，雌性，上海必凯实验动物有限公司，动物生产许可证号(SCXK(沪)2013-0006 N0.311620400001794)。21 tumor-bearing nude mice/example, female, Shanghai Bikai Experimental Animal Co., Ltd., animal production license number (SCXK (Shanghai) 2013-0006 No. 311620400001794).
2.3药物配制：2.3 Drug preparation:
0.5％HPMC(1％Tween 80)，超声溶解，配制为澄清溶液或均一混悬液。0.5% HPMC (1% Tween 80), ultrasonically dissolved, prepared as a clear solution or a homogeneous suspension.
2.4给药：2.4 Administration:
荷瘤裸鼠，雌性；禁食一夜后分别p.o.，剂量为30mg/kg，给药体积10mL/kg。Tumor-bearing nude mice, female; p.o. after fasting overnight, the dose is 30mg/kg, and the administration volume is 10mL/kg.
2.5样品采集：2.5 Sample collection:
荷瘤裸鼠给药前和给药后，在0、1、2、4、6、8、16和24小时，采用眼眶采血0.1mL，置于EDTA-K 2试管中，4℃6000rpm离心6min分离血浆，解剖后取肿瘤组织称重，于-80℃保存。 Before and after administration of tumor-bearing nude mice, at 0, 1, 2, 4, 6, 8, 16 and 24 hours, 0.1 mL of orbital blood was collected, placed in EDTA-K 2 test tubes, and centrifuged at 6000 rpm at 4 °C for 6 min Plasma was separated, tumor tissue was weighed after dissection, and stored at -80°C.
2.6样品处理：2.6 Sample handling:
1)肿瘤组织按1：3加入甲醇水匀浆后离心取上清，肿瘤匀浆液上清和血浆样品均取40μL加入160μL乙腈沉淀，混合后3500×g离心5～20分钟。1) Tumor tissue was homogenized by adding methanol water at a ratio of 1:3 and centrifuged to obtain the supernatant. Take 40 μL of the tumor homogenate supernatant and plasma samples and add 160 μL of acetonitrile for precipitation. After mixing, centrifuge at 3500×g for 5-20 minutes.
2)取处理后上清溶液100μL进行LC/MS/MS分析待测化合物的浓度。2) Take 100 μL of the treated supernatant solution and analyze the concentration of the compound to be tested by LC/MS/MS.
2.7液相分析2.7 Liquid phase analysis
●液相条件：Shimadzu LC-20AD泵●Liquid phase conditions: Shimadzu LC-20AD pump
●质谱条件:AB Sciex API 4000质谱仪●Mass spectrometer conditions: AB Sciex API 4000 mass spectrometer
●色谱柱：phenomenex Gemiu 5μm C18 50×4.6mm●Column: phenomenonex Gemiu 5μm C18 50×4.6mm
●移动相：A液为0.1％甲酸水溶液，B液为乙腈●Mobile phase: liquid A is 0.1% formic acid aqueous solution, liquid B is acetonitrile
●流速：0.8mL/min●Flow rate: 0.8mL/min
●洗脱时间：0-4.0分钟，洗脱液如下：●Elution time: 0-4.0 minutes, the eluent is as follows:
Figure PCTCN2022102195-appb-000056
Figure PCTCN2022102195-appb-000056
Figure PCTCN2022102195-appb-000057
Figure PCTCN2022102195-appb-000057
3.试验结果与分析3. Test results and analysis
药代动力学主要参数用WinNonlin 8.2计算得到，荷瘤裸鼠药代实验结果见下表：The main parameters of pharmacokinetics were calculated by WinNonlin 8.2, and the results of pharmacokinetic experiments on tumor-bearing nude mice are shown in the table below:
表15本发明实施例化合物在荷瘤裸鼠体内血浆/肿瘤中的浓度结果Table 15 Concentration results of the compounds of the embodiments of the present invention in plasma/tumor in tumor-bearing nude mice
Figure PCTCN2022102195-appb-000058
Figure PCTCN2022102195-appb-000058
4.实验结论：4. Experimental conclusion:
以上数据显示：本发明实施例化合物在小鼠肿瘤内的药物浓度高于血液药物浓度。The above data show that the drug concentration of the compound of the embodiment of the present invention in the mouse tumor is higher than the blood drug concentration.
测试例8血浆蛋白结合率实验Test example 8 plasma protein binding rate experiment
1.实验目的：1. Purpose of the experiment:
本实验方法的目的是检测实施例化合物在血浆中的血浆蛋白结合情况。The purpose of this experimental method is to detect the plasma protein binding of the compounds of the examples in plasma.
2.实验仪器及材料：2. Experimental instruments and materials:
超高效液相串联质谱联用联用仪，冷冻离心机，涡旋仪，电热恒温振荡水槽，移液器，连续加液器、96孔板、组织匀浆机(组织样品分析时使用)、加入内标的乙腈溶液、空白基质(血浆、尿液或组织匀浆液等)Ultra-high performance liquid phase tandem mass spectrometry coupled instrument, refrigerated centrifuge, vortex instrument, electric heating constant temperature oscillating water tank, pipette, continuous liquid adder, 96-well plate, tissue homogenizer (used for tissue sample analysis), Add internal standard acetonitrile solution, blank matrix (plasma, urine or tissue homogenate, etc.)
3.实验步骤：3. Experimental steps:
3.1准备血浆3.1 Preparation of plasma
于室温或37℃水浴解冻冷冻的血浆，3500rpm离心5min，取上清。Thaw the frozen plasma at room temperature or in a water bath at 37°C, centrifuge at 3500rpm for 5min, and take the supernatant.
3.2配制反应终止液3.2 Preparation of reaction termination solution
用含有内标的乙腈做终止液，储存在2-8℃冰箱。Use acetonitrile containing an internal standard as a stop solution and store in a refrigerator at 2-8°C.
3.3配制化合物工作液3.3 Preparation of compound working solution
化合物的工作液配制：用DMSO稀释储备液至终浓度1mM。Compound working solution preparation: dilute the stock solution with DMSO to a final concentration of 1 mM.
3.4配制血浆溶液3.4 Preparation of plasma solution
取2.5μL化合物工作液加入到497.5μL空白血浆中，终浓度为5μM，震荡混匀。Take 2.5 μL compound working solution and add to 497.5 μL blank plasma, the final concentration is 5 μM, shake and mix.
3.5平衡透析3.5 Equilibrium dialysis
1)准备平衡透析装置，将检测膜装置放入平衡透析96孔板中；1) Prepare the equilibrium dialysis device, and put the detection membrane device into the equilibrium dialysis 96-well plate;
2)在膜内加入200μL配制好的血浆溶液，n＝2；2) Add 200 μL of the prepared plasma solution into the membrane, n=2;
3)另取5μL血浆溶液，用45μL相同种属的空白血浆稀释10倍后，加入200μL含有内标的乙腈终止液，储存于-20℃冰箱；3) Take another 5 μL of plasma solution, dilute it 10 times with 45 μL of the same species of blank plasma, add 200 μL of acetonitrile stop solution containing internal standard, and store it in a -20°C refrigerator;
4)在膜外加入350μL透析液(100mM磷酸缓冲液)；4) Add 350 μL of dialysate (100 mM phosphate buffer) outside the membrane;
5)将透析板密封好，放入37℃水浴锅内孵育5小时；5) Seal the dialysis plate and place it in a water bath at 37°C for 5 hours;
6)透析结束后，自膜内样品孔分别取出5μL，用45μL相同种属的空白血浆稀释10倍；自膜外样品孔分别取出50μL透析液，加入200μL有内标的乙腈终止液；6) After the dialysis, take 5 μL from the sample holes inside the membrane and dilute 10 times with 45 μL of the same species of blank plasma; take 50 μL of the dialysate from the sample holes outside the membrane and add 200 μL of acetonitrile stop solution with internal standard;
7)样品离心后取上清液；7) Take the supernatant after the sample is centrifuged;
8)LC-MS分析。8) LC-MS analysis.
4.色谱条件：4. Chromatographic conditions:
仪器：岛津LC-30AD；Instrument: Shimadzu LC-30AD;
色谱柱：
Figure PCTCN2022102195-appb-000059
C18(50*4.6mm，5μm粒径)； Column:
Figure PCTCN2022102195-appb-000059
C18 (50*4.6mm, 5μm particle size);
流动相：A：0.1％甲酸溶液，B：甲醇；Mobile phase: A: 0.1% formic acid solution, B: methanol;
冲洗梯度：0.2～1.6min 5％A到95％A，3.0～3.1min 95％A到5％A；Washing gradient: 0.2～1.6min 5%A to 95%A, 3.0～3.1min 95%A to 5%A;
运行时间：4.0min。Running time: 4.0min.
5.质谱条件：5. Mass spectrometry conditions:
仪器：API5500型液相色谱质谱联用仪，美国AB Sciex公司；Instrument: API5500 liquid chromatography mass spectrometry, AB Sciex, USA;
离子源：电喷雾离子化源(ESI)；Ion source: electrospray ionization source (ESI);
干燥气体：N 2，温度500℃； Drying gas: N 2 , temperature 500°C;
电喷雾电压：5000V；Electrospray voltage: 5000V;
检测方式：正离子检测；Detection method: positive ion detection;
扫描方式：选择反应监测(MRM)方式。Scanning method: select the reaction monitoring (MRM) method.
6.实验结果：如表16所示：6. Experimental results: as shown in Table 16:
表16实施例化合物血浆蛋白结合率结果Table 16 Example compound plasma protein binding rate result
Figure PCTCN2022102195-appb-000060
Figure PCTCN2022102195-appb-000060
7.实验结论：7. Experimental conclusion:
以上数据显示：本发明实施例化合物显示出高血浆蛋白结合率。The above data show that: the compound of the embodiment of the present invention shows a high plasma protein binding rate.
测试例9 MC38-hPDL1移植瘤模型上对肿瘤的抑制实验Test Example 9 Tumor inhibition experiment on MC38-hPDL1 transplanted tumor model
1.实验目的：1. Purpose of the experiment:
评价受试化合物对MC38-hPDL1细胞C57小鼠皮下移植瘤的抗肿瘤活性。To evaluate the antitumor activity of the test compound on MC38-hPDL1 cell subcutaneous transplanted tumor of C57 mice.
2.实验仪器与试剂：2. Experimental instruments and reagents:
2.1仪器：2.1 Instruments:
超净工作台(BSC-1300II A2，上海博讯实业有限公司医疗设备厂)；Ultra-clean workbench (BSC-1300II A2, Shanghai Boxun Industrial Co., Ltd. Medical Equipment Factory);
CO 2培养箱(311，Thermo)； CO incubator (311, Thermo);
离心机(Centrifuge 5720R，Eppendorf)；Centrifuge (Centrifuge 5720R, Eppendorf);
全自动细胞计数仪(Countess II，Life)；Automatic cell counter (Countess II, Life);
移液器(10-20μL，Eppendorf)；Pipette (10-20 μL, Eppendorf);
显微镜(TS100，尼康)；Microscope (TS100, Nikon);
游标卡尺(500-196，日本三丰)；Vernier caliper (500-196, Mitutoyo, Japan);
细胞培养瓶(T25/T75/T225，Corning)。Cell culture flasks (T25/T75/T225, Corning).
2.2试剂：2.2 Reagents:
DMEM(11995-065，Gibco)；DMEM (11995-065, Gibco);
胎牛血清(FBS)(10099-141，Gibco)；Fetal bovine serum (FBS) (10099-141, Gibco);
磷酸盐缓冲液(PBS)(10010-023，Gibco)。Phosphate buffered saline (PBS) (10010-023, Gibco).
2.3受试化合物：2.3 Test compound:
本发明实施例化合物，自制。The compound of the embodiment of the present invention is self-made.
3.实验操作：3. Experimental operation:
从细胞库中取出MC38-hPDL1细胞，复苏后加入DMEM培养基中(DMEM+10％FBS)置于CO 2培养箱中培养(培养箱温度为37℃，CO 2浓度为5％)，待细胞数量扩增到体内接种所需数量时，收集MC38-hPDL1细胞。用全自动细胞计数仪计数，根据计数结果用PBS重悬细胞，制成细胞悬液(密度1×10 6/mL)，置于冰盒中待用。 Take out MC38-hPDL1 cells from the cell bank, add DMEM medium (DMEM+10% FBS) after recovery, and place them in a CO 2 incubator for culture (incubator temperature is 37°C, CO 2 concentration is 5%). When the number expanded to the required number for inoculation in vivo, MC38-hPDL1 cells were harvested. Count with an automatic cell counter, and resuspend the cells with PBS according to the counting results to make a cell suspension (density 1×10 6 /mL), and place in an ice box until use.
使用6-8周龄雌性C57小鼠，体重约为18-22克。小鼠饲养于SPF级动物房中，单笼饲养，每笼4-5只小鼠。所有的笼子、垫料和水在使用前进行高温消毒，所有动物均可自由饮食、饮水。实验开始前用一次性大小鼠通用耳标标记C57小鼠，接种前用75％酒精消毒接种部位皮肤，每只小鼠在右后背皮下接种0.1mL(含1*10 5个细胞)MC38-hPDL1细胞。当肿瘤体积达到40-180mm 3时开始分组给药，每组8只。各受试化合物每天口服给药2次、持续14天。每周测量肿瘤体积、称量小鼠体重各2次，并计算肿瘤TGI(％)。 Use 6-8 week old female C57 mice weighing approximately 18-22 g. Mice were raised in SPF grade animal room, and raised in single cages, with 4-5 mice per cage. All cages, litter and water were sterilized before use, and all animals had free access to food and water. Before the experiment started, the C57 mice were marked with a disposable ear tag for rats and rats, and the skin of the inoculation site was disinfected with 75% alcohol before inoculation, and each mouse was subcutaneously inoculated with 0.1 mL (containing 1*10 5 cells) of MC38- hPDL1 cells. When the tumor volume reached 40-180mm 3 , group administration began, with 8 rats in each group. Each test compound was orally administered twice a day for 14 days. Tumor volume was measured and mouse body weight was weighed twice a week, and tumor TGI (%) was calculated.
4.数据处理：4. Data processing:
肿瘤体积(mm 3)，计算公式为：V＝0.5*D*d*d，其中D和d分别是肿瘤的长径和短径。 Tumor volume (mm 3 ), the calculation formula is: V=0.5*D*d*d, where D and d are the long and short diameters of the tumor, respectively.
TGI(％)的计算：Calculation of TGI(%):
当肿瘤无消退时，TGI(％)＝[(1-(某处理组给药结束时平均瘤体积－该处理组开始给药时平均瘤体积))/(溶剂对照组治疗结束时平均瘤体积－溶剂对照组开始治疗时平均瘤体积)]×100％；When the tumor does not regress, TGI (%)=[(1-(average tumor volume at the end of administration of a certain treatment group-average tumor volume at the beginning of administration of this treatment group))/(average tumor volume at the end of treatment of the solvent control group - Average tumor volume at the beginning of treatment in solvent control group)]×100%;
当肿瘤有消退时，TGI(％)＝[1-(某处理组给药结束时平均瘤体积－该处理组开始给药时平均瘤体积)/该处理组开始给药时平均瘤体积]×100％。When the tumor has regressed, TGI (%)=[1-(average tumor volume at the end of administration of a treatment group-average tumor volume at the beginning of administration of this treatment group)/average tumor volume at the beginning of administration of this treatment group]× 100%.
5.实验结果：5. Experimental results:
试验结果如下表17：The test results are shown in Table 17 below:
表17化合物对移植瘤小鼠药效参数Table 17 compound's drug effect parameters on transplanted tumor mice
Figure PCTCN2022102195-appb-000061
Figure PCTCN2022102195-appb-000061
备注：括号中的数据表示，该实施例对应Vehicle QD x 2w组(即对照组)相应时间的肿瘤体积Remarks: The data in brackets indicates that this embodiment corresponds to the tumor volume of the Vehicle QD x 2w group (i.e. the control group) at the corresponding time
6.试验结论6. Test conclusion
以上数据显示：本发明实施例化合物对MC38-hPDL1细胞C57小鼠皮下移植瘤具有较强的抑制作用。The above data show that the compounds of the examples of the present invention have a strong inhibitory effect on MC38-hPDL1 cell subcutaneous transplanted tumors in C57 mice.
测试例10实施例4化合物SD大鼠重复灌胃14天毒性试验Test Example 10 Example 4 Compound SD rats repeated gavage toxicity test for 14 days
1.实验目的1. Purpose of the experiment
本研究的目的是考察实施例4化合物重复灌胃给予SD大鼠14天后可能出现的毒性反应。The purpose of this study is to investigate the possible toxic reaction of the compound of Example 4 given to SD rats after repeated gavage for 14 days.
2.实验材料和仪器2. Experimental Materials and Instruments
2.1供试品2.1 Test product
供试品：N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺(实施例4化合物)Test article: N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine-2 -yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazole A[4,5-c]pyridine-2-carboxamide (compound of Example 4)
2.2溶媒2.2 Solvent
名称：含1％(v/v)Tween 80的0.5％(w/v)HPMC K4M水溶液Name: 0.5% (w/v) HPMC K4M aqueous solution containing 1% (v/v) Tween 80
2.3动物信息2.3 Animal Information
种属&品系：SD大鼠Species & strain: SD rats
动物等级：SPF级Animal grade: SPF grade
动物数量和性别：8雄8雌/组Number and sex of animals: 8 males and 8 females/group
2.4仪器2.4 Instruments
Figure PCTCN2022102195-appb-000062
2120系列血液分析仪，Sysmex CS-2000i全自动凝血分析仪，TBA-120FR全自动生化分析仪。
Figure PCTCN2022102195-appb-000062
2120 series blood analyzer, Sysmex CS-2000i automatic coagulation analyzer, TBA-120FR automatic biochemical analyzer.
3.实验方法3. Experimental method
重复灌胃给予SD大鼠实施例4化合物，每天给药1次，连续给药14天，定期对大鼠临床观察、体重、耗食量、临床病理、毒代动力学、大体及组织病理学等指标进行检测，考察大鼠连续给药实施例4化合物14天后可能出现的毒性反应及代谢情况。Repeated gavage administration of the compound of Example 4 to SD rats was administered once a day for 14 consecutive days, and the clinical observations, body weight, food consumption, clinical pathology, toxicokinetics, general and histopathology of the rats were regularly monitored. Indicators were detected to investigate the possible toxic reaction and metabolism in rats after 14 days of continuous administration of the compound of Example 4.
4.试验数据4. Test data
4.1濒死/死亡4.1 Dying/Death
15、50、100mg/kg给药剂量组未见动物死亡/濒死。There was no death/moribundity of animals in the 15, 50, 100 mg/kg dose groups.
4.2毒代动力学4.2 Toxicokinetics
在15、50、100mg/kg剂量下，动物体内实施例4化合物的平均暴露量(AUC last)均随剂量增加而增大；动物体内暴露量增大的比率基本接近于剂量增加的比率；雌雄动物实施例4化合物的暴露量无明显性别差异。重复给药14天与首次给药相比，各剂量组动物体内实施例4化合物未见明显蓄积。 Under 15,50,100mg/kg dosage, the average exposure amount (AUC last ) of the compound of Example 4 in the animal body all increases with dosage; There was no significant gender difference in the exposure of the compound of Example 4 in animals. After 14 days of repeated administration, compared with the first administration, there was no significant accumulation of the compound of Example 4 in the animals of each dose group.
4.3临床观察4.3 Clinical Observation
100mg/kg剂量组1/5雄性动物偶见软便，15、50mg/kg剂量组未见异常。1/5 male animals in the 100mg/kg dose group occasionally had soft stools, and no abnormalities were found in the 15 and 50mg/kg dose groups.
4.4体重及食量4.4 Body weight and food intake
100mg/kg剂量组W1和W2体重增长减缓、食量降低，50mg/kg剂量组雌性动物W2体重增长减缓；15mg/kg剂量组未见异常。100mg/kg dose group W1 and W2 body weight growth slowed down, food intake decreased, 50mg/kg dose group female animals W2 body weight growth slowed down; 15mg/kg dose group no abnormalities.
4.5血细胞计数和凝血功能4.5 Blood cell count and coagulation function
100mg/kg剂量组Neut、FIB升高；15和50mg/kg剂量组雌性动物FIB升高。Neut and FIB increased in 100mg/kg dose group; FIB increased in female animals in 15 and 50mg/kg dose groups.
4.6血液生化4.6 Blood biochemistry
100mg/kg剂量组Alb降低；15、50mg/kg剂量组未见异常。100mg/kg dose group Alb decreased; 15, 50mg/kg dose groups no abnormalities.
4.7病理4.7 Pathology
15、50、100mg/kg剂量组动物大体观察未见异常，未进行组织病理学检查。Animals in the 15, 50, and 100 mg/kg dose groups had no abnormalities in general observation, and no histopathological examination was performed.
5.实验结论5. Experimental conclusion
实施例4化合物以15、50、100mg/kg的剂量重复灌胃给予SD大鼠，每天 给药1次，连续给药14天，最大耐受剂量(MTD)为100mg/kg。The compound of Example 4 was administered to SD rats repeatedly by intragastric administration at doses of 15, 50, and 100 mg/kg, once a day, for 14 consecutive days, and the maximum tolerated dose (MTD) was 100 mg/kg.
化合物的晶型研究Crystal Form Study of Compounds
本领域普通技术人员所熟知的是，上述实施例化合物被证明对CHO-PDL1/Jurkat-PD1细胞通路有较好的抑制活性，其晶型往往会具有同样的药理药效活性。在此基础上，发明人进一步研究了相应化合物晶型的理化性质，但下述具体晶型的制备与表征并不代表对本发明保护范围范围的限定，本领域普通技术人员可以以本发明为基础，通过常规的析晶手段获得本发明化合物更多的晶体，这些晶体均为本发明所保护的方案。具体如下：Those of ordinary skill in the art are well aware that the compounds of the above examples have been proven to have good inhibitory activity on the CHO-PDL1/Jurkat-PD1 cell pathway, and their crystal forms often have the same pharmacological activity. On this basis, the inventors further studied the physical and chemical properties of the corresponding compound crystal forms, but the preparation and characterization of the following specific crystal forms do not represent a limitation to the scope of protection of the present invention. Those of ordinary skill in the art can use the present invention as a basis , Obtain more crystals of the compound of the present invention through conventional crystallization means, and these crystals are all schemes protected by the present invention. details as follows:
1.实验仪器1. Experimental equipment
1.1物理化学检测仪器的一些参数1.1 Some parameters of physical and chemical detection instruments
Figure PCTCN2022102195-appb-000063
Figure PCTCN2022102195-appb-000063
1.2仪器和液相分析条件1.2 Instruments and liquid phase analysis conditions
Figure PCTCN2022102195-appb-000064
Figure PCTCN2022102195-appb-000064
Figure PCTCN2022102195-appb-000065
Figure PCTCN2022102195-appb-000065
2.实施例4化合物自由碱晶型A～D的制备2. Preparation of free base crystal forms A to D of the compound of Example 4
2.1自由碱晶型A的制备2.1 Preparation of free base crystal form A
加入粗产品DCM浓缩液与异丙醇(5V)，控制温度40-45℃继续浓缩至1-2V。重复该操作一次。补加异丙醇至5-6V，保持温度40-45℃搅拌10-20min。保持温度35-45℃热过滤，异丙醇(1-2V)漂洗。控制温度40-45℃，浓缩滤液至5V，搅拌10-30min，体系澄清。缓慢降至15-20℃，降温时间约1-2小时(自然降温亦可)。控制该温度搅拌16-20小时，能观察到有固体逐渐析出。过滤，异丙醇(1-2V)漂洗滤饼，得浅棕色固体。油泵抽干得到产品固体。得到的固体检测XRPD为晶型A。经检测分析，其有如图1所示的XRPD图及如图2所示的DSC图。Add the crude product DCM concentrate and isopropanol (5V), control the temperature at 40-45°C and continue to concentrate to 1-2V. Repeat this operation once. Add isopropanol to 5-6V, keep stirring at 40-45°C for 10-20min. Keep the temperature at 35-45°C for hot filtration and rinse with isopropanol (1-2V). Control the temperature at 40-45°C, concentrate the filtrate to 5V, stir for 10-30min, and the system is clarified. Slowly drop to 15-20°C, and the cooling time is about 1-2 hours (natural cooling is also acceptable). Control the temperature and stir for 16-20 hours, and it can be observed that a solid gradually precipitates out. Filter and rinse the filter cake with isopropanol (1-2V) to obtain a light brown solid. The oil pump is drained to obtain the product solid. The obtained solid detection XRPD is Form A. After detection and analysis, it has an XRPD pattern as shown in FIG. 1 and a DSC pattern as shown in FIG. 2 .
2.2自由碱晶型B的制备2.2 Preparation of free base crystal form B
加入粗产品DCM浓缩液(28g起始物料反应而得)与无水乙醇(5V)，控制温度40-45℃继续浓缩至1-2V。重复该操作一次。补加无水乙醇至5-6V，保持温度40-45℃搅拌10-20min。保持温度35-45℃热过滤，无水乙醇(1-2V)漂洗。控制温度40-45℃，浓缩滤液至3V，搅拌10-30min，体系基本澄清；控制温度40-45℃，滴加甲基叔丁基醚(112ml，4V)(备注：滴加速度不可过快，否则易形成油状物)约半小时滴加完毕。保温20-30min；缓慢降至15-20℃，降温时间约1-2小时(自然降温亦可)。控制该温度搅拌16-20小时，能观察到有固体逐渐析出。过滤，以MTBE：EtOH＝2：1(1-2V)漂洗滤饼，得浅棕色固体。得到的固体检测XRPD为晶型B。经检测分析，其有如图3所示的XRPD图及如图4所示的DSC图。Add the crude product DCM concentrate (obtained by reacting 28g of starting material) and absolute ethanol (5V), control the temperature at 40-45°C and continue to concentrate to 1-2V. Repeat this operation once. Add absolute ethanol to 5-6V, keep stirring at 40-45°C for 10-20min. Keep the temperature at 35-45°C for hot filtration and rinse with absolute ethanol (1-2V). Control the temperature at 40-45°C, concentrate the filtrate to 3V, stir for 10-30min, the system is basically clear; control the temperature at 40-45°C, add methyl tert-butyl ether (112ml, 4V) dropwise (note: the rate of addition should not be too fast, Otherwise it is easy to form an oily substance) about half an hour to complete the dropwise addition. Keep warm for 20-30 minutes; slowly drop to 15-20°C, and the cooling time is about 1-2 hours (natural cooling is also acceptable). Control the temperature and stir for 16-20 hours, and it can be observed that a solid gradually precipitates out. Filter and rinse the filter cake with MTBE:EtOH=2:1 (1-2V) to obtain a light brown solid. The obtained solid detection XRPD is crystal form B. After detection and analysis, it has an XRPD pattern as shown in FIG. 3 and a DSC pattern as shown in FIG. 4 .
2.3自由碱晶型C的制备2.3 Preparation of free base crystal form C
称量约50mg的游离碱B，加入1mL甲酸乙酯室温溶清，在室温下加入1mL环己烷混浊成油，室温搅拌至固体析出。最后固体快速离心，去除上清液，40℃真空干燥，得到的固体检测XRPD为晶型C。经检测分析，其有如图5所示的XRPD图及如图6所示的DSC图。Weigh about 50 mg of free base B, add 1 mL of ethyl formate to dissolve at room temperature, add 1 mL of cyclohexane at room temperature to become cloudy, and stir at room temperature until solid precipitates. Finally, the solid was centrifuged quickly, the supernatant was removed, and vacuum-dried at 40° C., the obtained solid was found to be crystal form C by XRPD. After detection and analysis, it has an XRPD pattern as shown in FIG. 5 and a DSC pattern as shown in FIG. 6 .
2.4自由碱晶型D的制备2.4 Preparation of free base crystal form D
称量约10mg的游离碱晶型C，加入0.1mL乙酸乙酯于40℃下打浆2小时，然后室温打浆12小时，最后得到的固体检测XRPD为晶型D。经检测分析，其有如图7所示的XRPD图及如图8所示的DSC图。Weigh about 10mg of free base crystal form C, add 0.1mL ethyl acetate and beat at 40°C for 2 hours, then beat at room temperature for 12 hours, and the final solid obtained is crystal form D by XRPD. After detection and analysis, it has an XRPD pattern as shown in FIG. 7 and a DSC pattern as shown in FIG. 8 .
3.稳定晶型确认实验3. Stable crystal form confirmation experiment
3.1实验目的：3.1 Purpose of the experiment:
通过晶型打浆实验稳定性考察实验找到实施例4化合物比较稳定的晶型。A relatively stable crystal form of the compound of Example 4 was found through the crystal form beating experiment stability investigation experiment.
3.2实验方案：3.2 Experimental scheme:
选择有一定溶解度的有机溶剂、水，将不同晶型悬浮于溶剂体系中，一定温度下搅拌打浆最后将固体处理，测定固体的XRPD并进行比较。Choose an organic solvent and water with a certain solubility, suspend different crystal forms in the solvent system, stir and beat at a certain temperature, and finally treat the solid, measure the XRPD of the solid and compare them.
3.3实验结果：3.3 Experimental results:
称取自由碱晶型A约20mg样品，加入0.1mL溶剂，于室温打浆。具体结果如下表18所示。Weigh about 20mg sample of free base crystal form A, add 0.1mL solvent, and beat at room temperature. The specific results are shown in Table 18 below.
表18自由碱稳定晶型确认实验结果Table 18 Confirmation experiment results of free base stable crystal form
Figure PCTCN2022102195-appb-000066
Figure PCTCN2022102195-appb-000066
Figure PCTCN2022102195-appb-000067
Figure PCTCN2022102195-appb-000067
实验结论：从DSC数据中可知，晶型B为最稳定晶型。Experimental conclusion: From the DSC data, it can be seen that the crystal form B is the most stable crystal form.
4.固体稳定性实验4. Solid stability experiment
4.1实验目的：4.1 Purpose of the experiment:
考察实施例4化合物自由碱晶型B在高温(60℃)，室温高湿(RH＝92.5％)及高温高湿(50℃，RH＝75％)三个条件下，化合物的物理化学稳定性，为晶型筛选与化合物贮存提供依据。Investigate the physical and chemical stability of the compound free base crystal form B of Example 4 under the conditions of high temperature (60°C), room temperature and high humidity (RH=92.5%) and high temperature and high humidity (50°C, RH=75%) , to provide a basis for crystal form screening and compound storage.
4.2实验方案：4.2 Experimental scheme:
实验中称取约5mg自由碱晶型B于20mL的小瓶中，分别置于高温(60℃)，室温高湿(RH＝92.5％)及高温高湿(50℃，RH＝75％)三个条件下，5天、10天、20天、30天取样，用HPLC外标法测定含量，并采用色谱峰面积归一化法计算有关物质的变化。In the experiment, about 5mg of free base crystal form B was weighed into a 20mL vial, and placed in high temperature (60°C), room temperature and high humidity (RH=92.5%) and high temperature and high humidity (50°C, RH=75%) three Under the same conditions, samples were taken at 5 days, 10 days, 20 days, and 30 days, and the content was determined by the HPLC external standard method, and the change of related substances was calculated by the chromatographic peak area normalization method.
4.3实验结果：4.3 Experimental results:
实施例4化合物游离碱晶型B固体稳定性实验结果如下表19所示：The solid stability test results of the free base crystal form B of the compound of Example 4 are shown in Table 19 below:
表19自由碱晶型B固体稳定性实验结果Table 19 Solid stability test results of free base crystal form B
Figure PCTCN2022102195-appb-000068
Figure PCTCN2022102195-appb-000068
实验结论：自由碱晶型B在高温高湿条件下稳定性很好，且最大单杂增加小于0.1％。Experimental conclusion: the free base crystal form B has good stability under high temperature and high humidity conditions, and the maximum single-heterogeneity increase is less than 0.1%.
5.引湿性实验5. Moisture test
5.1实验目的5.1 Purpose of experiment
考察实施例4化合物自由碱晶型B在不同相对湿度条件下的引湿性，为化合物晶型筛选与贮存提供依据。The hygroscopicity of the free base crystal form B of the compound of Example 4 was investigated under different relative humidity conditions to provide a basis for the screening and storage of the compound crystal form.
5.2实验方案：5.2 Experimental scheme:
将化合物自由碱晶型B置不同相对湿度的饱和水蒸气中，使化合物与水蒸气达到动态平衡，并计算平衡后化合物吸湿增重的百分数。The free base crystal form B of the compound is placed in saturated water vapor with different relative humidity, so that the compound and the water vapor reach a dynamic equilibrium, and the percentage of the weight gain of the compound after the equilibrium is calculated.
5.3实验结果：5.3 Experimental results:
自由碱晶型B在RH80％条件下吸湿增重约0.815％，略有引湿性。经0-95％相对湿度条件下吸湿与解吸湿循环1次，自由碱晶型B的XRPD谱图未发生改变，即晶型B未转晶。Under the condition of RH80%, the free base crystal form B has a hygroscopic weight gain of about 0.815%, which is slightly hygroscopic. After one cycle of moisture absorption and desorption under the condition of 0-95% relative humidity, the XRPD spectrum of the free base crystal form B does not change, that is, the crystal form B is not transformed.
6.不同介质中溶解度实验6. Solubility experiments in different media
6.1实验目的6.1 Purpose of experiment
比较实施例4化合物自由碱晶型B在pH 1～pH 8缓冲液、水、人工模拟胃液(SGF)、禁食人工模拟肠液(FaSSIF)及非禁食人工模拟肠液(FeSSIF)等媒介中溶解度大小，为可成药性评估提供依据。Comparing the solubility of the free base crystal form B of the compound of Example 4 in media such as pH 1～pH 8 buffer solution, water, artificial simulated gastric juice (SGF), fasted artificial simulated intestinal fluid (FaSSIF) and non-fasting artificial simulated intestinal fluid (FeSSIF) The size provides a basis for the assessment of druggability.
6.2实验方案：6.2 Experimental scheme:
将约2mg自由碱晶型B混悬到不同介质中2小时，用HPLC，外标法测定化合物37℃下的热力学溶解度。About 2 mg of free base crystal form B was suspended in different media for 2 hours, and the thermodynamic solubility of the compound at 37°C was determined by HPLC and external standard method.
6.3实验结果：6.3 Experimental results:
不同介质中溶解度实验结果如下表20所示：The solubility test results in different media are shown in Table 20 below:
表20自由碱晶型B不同介质中溶解度实验结果Table 20 Solubility test results in different media of free base crystal form B
溶解度/介质Solubility/medium 溶解度(mg/mL)Solubility (mg/mL)
pH1pH1 >1.640>1.640
pH2pH2 >1.052>1.052
pH3pH3 0.2000.200
pH4pH4 0.2860.286
pH5pH5 >1.375>1.375
pH6pH6 0.2360.236
pH7pH7 0.0400.040
pH8pH8 0.0000.000
FaSSIFFaSSIF 0.6350.635
FeSSIFFeSSIF >1.217>1.217
FaSSGFFaSSGF >1.298>1.298
H 2O H 2 O 0.0000.000
实验结论：自由碱晶型B在整个胃肠道包含的pH范围及环境下均具有很好的溶解度。Experimental conclusion: Free base crystal form B has good solubility in the pH range and environment of the entire gastrointestinal tract.

Claims (16)

  1. 通式(I)所示化合物的晶型，化合物结构如下：The crystal form of the compound shown in general formula (I), the compound structure is as follows:
    Figure PCTCN2022102195-appb-100001
    Figure PCTCN2022102195-appb-100001
    其中：in:
    L选自键、
    Figure PCTCN2022102195-appb-100002
    L is selected from the key,
    Figure PCTCN2022102195-appb-100002
    环A选自4-8元杂环基；优选4元、7元或8元杂环基；Ring A is selected from 4-8 membered heterocyclic groups; preferably 4-membered, 7-membered or 8-membered heterocyclic groups;
    更优选如下基团：More preferred are the following groups:
    Figure PCTCN2022102195-appb-100003
    Figure PCTCN2022102195-appb-100003
    R 1独立地选自选自氢、氘、卤素、羟基、氰基、羧基、醛基、氧代基、C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6羟烷基、C 3-8环烷基、-(CH 2) nR a、-(CH 2) nOR a、-OC(O)R a、-C(O)R a、-C(O)OR a、-C(O)NR aR b、-(CH 2) nNR aC(O)R b或-SO 2R a，其中所述的C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基、C 1-6烷氧基、C 1-6羟烷基和C 3-8环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-6烷基中的一个或多个取代基所取代； R is independently selected from hydrogen, deuterium, halogen, hydroxyl, cyano, carboxyl, aldehyde, oxo, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl , C 1-6 alkoxy, C 1-6 hydroxyalkyl, C 3-8 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O)R a , -C(O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a , wherein The C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl, C 1-6 alkoxy, C 1-6 hydroxyalkyl and C 3-8 cycloalkyl, any selected to be further substituted by one or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-6 alkyl;
    R a和R b各自独立地选自氢、氘、卤素、氨基、羟基、巯基、氰基、硝基、羧基、C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基或C 3-8环烷基，其中所述的C 1-6烷基、C 1-6氘代烷基、C 1-6卤代烷基和C 3-8环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-6烷基中的一个或多个取代基所取代； R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, carboxyl, C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 Haloalkyl or C 3-8 cycloalkyl, wherein said C 1-6 alkyl, C 1-6 deuterated alkyl, C 1-6 haloalkyl and C 3-8 cycloalkyl, optionally further One or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-6 alkyl;
    n为0、1或2；且n is 0, 1 or 2; and
    x为0、1或2。x is 0, 1 or 2.
  2. 根据权利要求1所述化合物的晶型，所述化合物的结构如通式(II)所示：The crystal form of the compound according to claim 1, the structure of the compound is as shown in general formula (II):
    Figure PCTCN2022102195-appb-100004
    Figure PCTCN2022102195-appb-100004
    其中：in:
    M为O、-NR 2或-CR 3R 4； M is O, -NR 2 or -CR 3 R 4 ;
    R 2选自氢、氘、醛基、C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基、C 1-3烷氧基、C 1-3羟烷基、C 3-6环烷基、-(CH 2) nR a、-(CH 2) nOR a、-OC(O)R a、-C(O)R a、-C(O)OR a、-C(O)NR aR b、-(CH 2) nNR aC(O)R b或-SO 2R a； R 2 is selected from hydrogen, deuterium, aldehyde, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 3-6 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O)R a , -C(O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a ;
    R 3和R 4各自独立地选自氢、氘、卤素、羟基、氰基、羧基、醛基、氧代基、C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基、C 1-3烷氧基、C 1-3羟烷基、C 3-6环烷基、-(CH 2) nR a、-(CH 2) nOR a、-OC(O)R a、-C(O)R a、-C(O)OR a、-C(O)NR aR b、-(CH 2) nNR aC(O)R b或-SO 2R a，其中所述的C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基、C 1-3烷氧基、C 1-3羟烷基和C 3-6环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-3烷基中的一个或多个取代基所取代； R 3 and R 4 are each independently selected from hydrogen, deuterium, halogen, hydroxyl, cyano, carboxyl, aldehyde, oxo, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 Haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl, C 3-6 cycloalkyl, -(CH 2 ) n R a , -(CH 2 ) n OR a , -OC(O) R a , -C(O)R a , -C(O)OR a , -C(O)NR a R b , -(CH 2 ) n NR a C(O)R b or -SO 2 R a , The C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl, C 1-3 alkoxy, C 1-3 hydroxyalkyl and C 3-6 cycloalkyl , optionally further substituted by one or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-3 alkyl;
    R a和R b各自独立地选自氢、氘、卤素、氨基、羟基、巯基、氰基、硝基、羧基、C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基或C 3-6环烷基，其中所述的C 1-3烷基、C 1-3氘代烷基、C 1-3卤代烷基和C 3-6环烷基，任选进一步被氘、卤素、羟基、氰基或C 1-3烷基中的一个或多个取代基所取代； R a and R b are each independently selected from hydrogen, deuterium, halogen, amino, hydroxyl, mercapto, cyano, nitro, carboxyl, C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 Haloalkyl or C 3-6 cycloalkyl, wherein said C 1-3 alkyl, C 1-3 deuterated alkyl, C 1-3 haloalkyl and C 3-6 cycloalkyl, optionally further One or more substituents in deuterium, halogen, hydroxyl, cyano or C 1-3 alkyl;
    n为0、1或2。n is 0, 1 or 2.
  3. 根据权利要求1或2所述化合物的晶型，所述化合物为：According to the crystal form of the compound according to claim 1 or 2, the compound is:
    N-(2,2'-二氯-3'-(6-甲氧基-5-((噁丁环-3-基氨基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((oxetan-3-ylamino)methyl)pyridin-2-yl)-[1,1'- Biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((3-氟吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-fluoroazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
    1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-基乙酸酯；1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-carbobalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine- 3-yl acetate;
    N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
    N-(3'-(5-((2-氧杂-6-氮杂螺[3.3]庚烷-6-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((2-Oxa-6-azaspiro[3.3]heptane-6-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]pyridine-2-carboxamide;
    N-(3'-(5-((5-氧杂-2-氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二 氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((5-oxa-2-azaspiro[3.4]octane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(6-甲氧基-5-((6-羰基-2,5-二氮杂螺[3.4]辛烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-carbonyl-2,5-diazaspiro[3.4]octane-2-yl)methyl) Pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5 -c] pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((3-羟基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-hydroxyazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-[1,1 '-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((3-羟基-3-甲基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-hydroxy-3-methylazetidin-1-yl)methyl)-6-methoxypyridin-2-yl) -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 - Formamide;
    1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺基)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-3-羧酸；1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-Carboxamido)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl)azetidine-3-carboxylic acid;
    (S)-1-((6-(2,2'-二氯-3'-(1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-碳杂草酰氨基<乙二酰氨基>)-[1,1'-联苯基]-3-基)-2-甲氧基吡啶-3-基)甲基)吖丁啶-2-羧酸；(S)-1-((6-(2,2'-dichloro-3'-(1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5 -c]pyridine-2-carboxalylamino<oxalylamino>)-[1,1'-biphenyl]-3-yl)-2-methoxypyridin-3-yl)methyl) Azetidine-2-carboxylic acid;
    (S)-N-(2,2'-二氯-3'-(5-((2-(羟甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；(S)-N-(2,2'-dichloro-3'-(5-((2-(hydroxymethyl)azetidin-1-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((3-羟基-3-(羟甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-hydroxy-3-(hydroxymethyl)azetidin-1-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide;
    N-(3'-(5-((3-乙酰氨基吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((3-acetylaminoazetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1, 1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
    N-(3'-(5-((3-(乙酰氨基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((3-(acetylaminomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 - Formamide;
    N-(2,2'-二氯-3'-(5-((3-(氰基甲基)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-(cyanomethyl)azetidin-1-yl)methyl)-6-methoxypyridin-2-yl) -[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2 - Formamide;
    N-(2,2'-二氯-3'-(5-((3-(二甲基氨基甲酰)吖丁啶-1-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((3-(dimethylcarbamoyl)azetidin-1-yl)methyl)-6-methoxypyridine-2- base)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine -2-formamide;
    N-(2,2'-二氯-3'-(6-甲氧基-5-((3-(甲基氨基甲酰)吖丁啶-1-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((3-(methylcarbamoyl)azetidin-1-yl)methyl)pyridin-2-yl )-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine- 2-formamide;
    N-(2,2'-二氯-3'-(5-((6-羟基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-hydroxy-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridine-2 -yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c] Pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6-methanol Oxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4 ,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((6-(环丙羰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-(cyclopropylcarbonyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl)-6 -Methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(6-甲氧基-5-((6-丙酰-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶 -2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-propionyl-2,6-diazaspiro[3.3]heptane-2-yl)methyl )pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(2,2,2-三氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(2,2,2-trifluoroacetyl)-2,6-diazaspiro[3.3 ]heptane-2-yl)methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7- Tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((6-(2,2-二氟乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-(2,2-difluoroacetyl)-2,6-diazaspiro[3.3]heptan-2-yl) Methyl)-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro -1H-imidazo[4,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((6-(2-氰基乙酰基)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-(2-cyanoacetyl)-2,6-diazaspiro[3.3]heptane-2-yl)methyl )-6-methoxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H - imidazo[4,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(6-甲氧基-5-((6-(甲磺酰)-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(6-methoxy-5-((6-(methylsulfonyl)-2,6-diazaspiro[3.3]heptane-2-yl )methyl)pyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo [4,5-c]pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((6-甲酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(2,2'-dichloro-3'-(5-((6-formyl-2,6-diazaspiro[3.3]heptan-2-yl)methyl)-6-methoxy Basepyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
    N-(3'-(5-((7-乙酰基-2,7-二氮杂螺[3.5]壬烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺N-(3'-(5-((7-acetyl-2,7-diazaspiro[3.5]nonan-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c]pyridine-2-carboxamide
    N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；N-(3'-(5-((6-acetyl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methoxypyridin-2-yl)- 2,2'-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4, 5-c] pyridine-2-carboxamide;
    N-(2,2'-二氯-3'-(5-((6-异丁酰-2,6-二氮杂螺[3.4]辛烷-2-基)甲基)-6-甲氧基吡啶-2-基)-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺；或N-(2,2'-dichloro-3'-(5-((6-isobutyryl-2,6-diazaspiro[3.4]octane-2-yl)methyl)-6-methanol Oxypyridin-2-yl)-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4 ,5-c]pyridine-2-carboxamide; or
    N-(3'-(5-((6-乙酰氨基-2-氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺。N-(3'-(5-((6-Acetamido-2-azaspiro[3.3]heptane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2 '-Dichloro-[1,1'-biphenyl]-3-yl)-1,5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c ] Pyridine-2-carboxamide.
  4. 根据权利要求1-3任一项所述化合物的晶型，其特征在于，所述晶型为N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型A～D；The crystal form of the compound according to any one of claims 1-3, wherein the crystal form is N-(3'-(5-((6-acetyl-2,6-diazaspiro[ 3.3] Heptane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1 , Crystal forms A to D of 5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
    所述晶型A的X-射线粉末衍射图谱在2θ为7.8±0.2°处具有衍射峰；或者在14.1±0.2°处具有衍射峰；或者在15.3±0.2°处具有衍射峰；或者在19.5±0.2°处具 有衍射峰；或者在12.0±0.2°处具有衍射峰；或者在12.4±0.2°处具有衍射峰；或者在13.2±0.2°处具有衍射峰；或者在14.6±0.2°处具有衍射峰；或者在21.2±0.2°处具有衍射峰；或者在22.0±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form A has a diffraction peak at 2θ of 7.8±0.2°; or has a diffraction peak at 14.1±0.2°; or has a diffraction peak at 15.3±0.2°; or has a diffraction peak at 19.5±0.2°; Diffraction peak at 0.2°; or diffraction peak at 12.0±0.2°; or diffraction peak at 12.4±0.2°; or diffraction peak at 13.2±0.2°; or diffraction peak at 14.6±0.2° or have a diffraction peak at 21.2±0.2°; or have a diffraction peak at 22.0±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
    所述晶型B的X-射线粉末衍射图谱在2θ为4.2±0.2°处具有衍射峰；或者在12.2±0.2°处具有衍射峰；或者在12.6±0.2°处具有衍射峰；或者在16.5±0.2°处具有衍射峰；或者在16.8±0.2°处具有衍射峰；或者在18.4±0.2°处具有衍射峰；或者在18.9±0.2°处具有衍射峰；或者在21.1±0.2°处具有衍射峰；或者在22.2±0.2°处具有衍射峰；或者在22.4±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form B has a diffraction peak at 2θ of 4.2±0.2°; or has a diffraction peak at 12.2±0.2°; or has a diffraction peak at 12.6±0.2°; or has a diffraction peak at 16.5±0.2°; Diffraction peak at 0.2°; or diffraction peak at 16.8±0.2°; or diffraction peak at 18.4±0.2°; or diffraction peak at 18.9±0.2°; or diffraction peak at 21.1±0.2° or have a diffraction peak at 22.2±0.2°; or have a diffraction peak at 22.4±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
    所述晶型C的X-射线粉末衍射图谱在2θ为11.4±0.2°处具有衍射峰；或者在12.5±0.2°处具有衍射峰；或者在21.1±0.2°处具有衍射峰；或者在23.0±0.2°处具有衍射峰；或者在26.1±0.2°处具有衍射峰；或者在26.6±0.2°处具有衍射峰；或者在13.0±0.2°处具有衍射峰；或者在14.0±0.2°处具有衍射峰；或者在14.7±0.2°处具有衍射峰；或者在15.8±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form C has a diffraction peak at 2θ of 11.4±0.2°; or has a diffraction peak at 12.5±0.2°; or has a diffraction peak at 21.1±0.2°; or has a diffraction peak at 23.0±0.2°; Diffraction peak at 0.2°; or diffraction peak at 26.1±0.2°; or diffraction peak at 26.6±0.2°; or diffraction peak at 13.0±0.2°; or diffraction peak at 14.0±0.2° or have a diffraction peak at 14.7±0.2°; or have a diffraction peak at 15.8±0.2°; preferably include any 2-5 of the above-mentioned diffraction peaks, or 3-5, or 3-6, or 3 -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places;
    所述晶型D的X-射线粉末衍射图谱在2θ为9.5±0.2°处具有衍射峰；或者在10.4±0.2°处具有衍射峰；或者在14.5±0.2°处具有衍射峰；或者在19.9±0.2°处具有衍射峰；或者在20.8±0.2°处具有衍射峰；或者在24.8±0.2°处具有衍射峰；或者在11.7±0.2°处具有衍射峰；或者在13.3±0.2°处具有衍射峰；或者在17.2±0.2°处具有衍射峰；或者在23.8±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处。The X-ray powder diffraction pattern of the crystal form D has a diffraction peak at 2θ of 9.5±0.2°; or has a diffraction peak at 10.4±0.2°; or has a diffraction peak at 14.5±0.2°; or has a diffraction peak at 19.9±0.2°; Diffraction peak at 0.2°; or diffraction peak at 20.8±0.2°; or diffraction peak at 24.8±0.2°; or diffraction peak at 11.7±0.2°; or diffraction peak at 13.3±0.2° or have a diffraction peak at 17.2±0.2°; or have a diffraction peak at 23.8±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3 of the above-mentioned diffraction peaks -8 places, or 5-8 places, or 6-8 places, more preferably including any 6, 7 or 8 places.
  5. 根据权利要求4所述化合物的晶型，其特征在于，The crystal form of the compound according to claim 4, characterized in that,
    晶型A的X-射线粉末衍射图谱至少包含位于2θ为7.8±0.2°、14.1±0.2°、15.3±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含19.5±0.2°、12.0±0.2°、12.4±0.2°、13.2±0.2°、14.6±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；The X-ray powder diffraction pattern of Form A contains at least one or more diffraction peaks at 2θ of 7.8±0.2°, 14.1±0.2°, and 15.3±0.2°, preferably two of them, more preferably three; Optionally, it may further include at least one of 19.5±0.2°, 12.0±0.2°, 12.4±0.2°, 13.2±0.2°, 14.6±0.2°, preferably 2, 3, 4 or 5 strip;
    晶型B的X-射线粉末衍射图谱至少包含位于2θ为4.2±0.2°、12.2±0.2°、12.6±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含16.5±0.2°、16.8±0.2°、18.4±0.2°、18.9±0.2°、21.1±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；The X-ray powder diffraction pattern of crystal form B contains at least one or more diffraction peaks at 2θ of 4.2±0.2°, 12.2±0.2°, and 12.6±0.2°, preferably two of them, more preferably three; Optionally, it may further include at least one of 16.5±0.2°, 16.8±0.2°, 18.4±0.2°, 18.9±0.2°, 21.1±0.2°, preferably 2, 3, 4 or 5 strip;
    晶型C的X-射线粉末衍射图谱至少包含位于2θ为11.4±0.2°、12.5±0.2°、21.1±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含23.0±0.2°、26.1±0.2°、26.6±0.2°、13.0±0.2°、14.0±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；The X-ray powder diffraction pattern of Form C contains at least one or more diffraction peaks located at 2θ of 11.4±0.2°, 12.5±0.2°, and 21.1±0.2°, preferably two of them, more preferably three; Optionally, it may further include at least one of 23.0±0.2°, 26.1±0.2°, 26.6±0.2°, 13.0±0.2°, 14.0±0.2°, preferably 2, 3, 4 or 5 strip;
    晶型D的X-射线粉末衍射图谱至少包含位于2θ为9.5±0.2°、10.4±0.2°、14.5±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包含三条；任选的，进一步还可以包含19.9±0.2°、20.8±0.2°、24.8±0.2°、11.7±0.2°、13.3±0.2°中的至少一条，优选包含其中2条、3条、4条或5条。The X-ray powder diffraction pattern of crystal form D contains at least one or more diffraction peaks located at 2θ of 9.5±0.2°, 10.4±0.2°, and 14.5±0.2°, preferably two of them, more preferably three; Optionally, it may further include at least one of 19.9±0.2°, 20.8±0.2°, 24.8±0.2°, 11.7±0.2°, 13.3±0.2°, preferably 2, 3, 4 or 5 strip.
  6. 根据权利要求5所述化合物的晶型，其特征在于，The crystal form of the compound according to claim 5, characterized in that,
    晶型A的X-射线粉末衍射图谱任选还包含位于2θ为21.2±0.2°、22.0±0.2°、22.6±0.2°、25.0±0.2°、28.3±0.2°、16.7±0.2°或24.5±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；The X-ray powder diffraction pattern of crystalline form A optionally further comprises a position at 2θ of 21.2±0.2°, 22.0±0.2°, 22.6±0.2°, 25.0±0.2°, 28.3±0.2°, 16.7±0.2° or 24.5±0.2 One or more diffraction peaks in °; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5 6, 7;
    晶型B的X-射线粉末衍射图谱任选还包含位于2θ为22.2±0.2°、22.4±0.2°、22.9±0.2°、23.6±0.2°、26.6±0.2°、13.8±0.2°或14.5±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；The X-ray powder diffraction pattern of the crystal form B optionally further comprises a position at 2θ of 22.2±0.2°, 22.4±0.2°, 22.9±0.2°, 23.6±0.2°, 26.6±0.2°, 13.8±0.2° or 14.5±0.2 One or more diffraction peaks in °; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5 6, 7;
    晶型C的X-射线粉末衍射图谱任选还包含位于2θ为14.7±0.2°、15.8±0.2°、18.0±0.2°、19.8±0.2°、23.6±0.2°、9.1±0.2°或16.7±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；The X-ray powder diffraction pattern of the crystal form C optionally further comprises a position at 2θ of 14.7±0.2°, 15.8±0.2°, 18.0±0.2°, 19.8±0.2°, 23.6±0.2°, 9.1±0.2° or 16.7±0.2 One or more diffraction peaks in °; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5 6, 7;
    晶型D的X-射线粉末衍射图谱任选还包含位于2θ为17.2±0.2°、23.8±0.2°、25.5±0.2°、27.1±0.2°、15.3±0.2°、15.8±0.2°或21.4±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处。The X-ray powder diffraction pattern of the crystal form D optionally further comprises a position at 2θ of 17.2±0.2°, 23.8±0.2°, 25.5±0.2°, 27.1±0.2°, 15.3±0.2°, 15.8±0.2° or 21.4±0.2 One or more diffraction peaks in °; preferably at least include any 2-3, or 4-5, or 6-7; more preferably, include any 2, 3, 4, 5 , 6, 7.
  7. 根据权利要求4所述化合物的晶型，其特征在于，The crystal form of the compound according to claim 4, characterized in that,
    晶型A的X-射线粉末衍射图谱任选包含位于2θ为7.8±0.2°、14.1±0.2°、15.3±0.2°、19.5±0.2°、12.0±0.2°、12.4±0.2°、13.2±0.2°、14.6±0.2°、21.2±0.2°、22.0±0.2°、22.6±0.2°、25.0±0.2°、28.3±0.2°、16.7±0.2°或24.5±0.2°处中的一处或多处衍射峰，The X-ray powder diffraction pattern of the crystal form A optionally comprises a position at 2θ of 7.8±0.2°, 14.1±0.2°, 15.3±0.2°, 19.5±0.2°, 12.0±0.2°, 12.4±0.2°, 13.2±0.2° , 14.6±0.2°, 21.2±0.2°, 22.0±0.2°, 22.6±0.2°, 25.0±0.2°, 28.3±0.2°, 16.7±0.2° or 24.5±0.2° at one or more diffraction peaks ,
    优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 places;
    晶型B的X-射线粉末衍射图谱任选包含位于2θ为4.2±0.2°、12.2±0.2°、12.6±0.2°、16.5±0.2°、16.8±0.2°、18.4±0.2°、18.9±0.2°、21.1±0.2°、22.2±0.2°、22.4±0.2°、22.9±0.2°、23.6±0.2°、26.6±0.2°、13.8±0.2°或14.5±0.2°处中的一处或 多处衍射峰，The X-ray powder diffraction pattern of the crystal form B optionally comprises 2θ at 4.2±0.2°, 12.2±0.2°, 12.6±0.2°, 16.5±0.2°, 16.8±0.2°, 18.4±0.2°, 18.9±0.2° , 21.1±0.2°, 22.2±0.2°, 22.4±0.2°, 22.9±0.2°, 23.6±0.2°, 26.6±0.2°, 13.8±0.2° or 14.5±0.2° at one or more diffraction peaks ,
    优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 places;
    晶型C的X-射线粉末衍射图谱任选包含位于2θ为11.4±0.2°、12.5±0.2°、21.1±0.2°、23.0±0.2°、26.1±0.2°、26.6±0.2°、13.0±0.2°、14.0±0.2°、14.7±0.2°、15.8±0.2°、18.0±0.2°、19.8±0.2°、23.6±0.2°、9.1±0.2°或16.7±0.2°处中的一处或多处衍射峰，The X-ray powder diffraction pattern of the crystal form C optionally comprises 2θ at 11.4±0.2°, 12.5±0.2°, 21.1±0.2°, 23.0±0.2°, 26.1±0.2°, 26.6±0.2°, 13.0±0.2° , 14.0±0.2°, 14.7±0.2°, 15.8±0.2°, 18.0±0.2°, 19.8±0.2°, 23.6±0.2°, 9.1±0.2° or 16.7±0.2° at one or more diffraction peaks ,
    优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 places;
    晶型D的X-射线粉末衍射图谱任选包含位于2θ为9.5±0.2°、10.4±0.2°、14.5±0.2°、19.9±0.2°、20.8±0.2°、24.8±0.2°、11.7±0.2°、13.3±0.2°、17.2±0.2°、23.8±0.2°、25.5±0.2°、27.1±0.2°、15.3±0.2°、15.8±0.2°或21.4±0.2°处中的一处或多处衍射峰，The X-ray powder diffraction pattern of the crystal form D optionally comprises 2θ at 9.5±0.2°, 10.4±0.2°, 14.5±0.2°, 19.9±0.2°, 20.8±0.2°, 24.8±0.2°, 11.7±0.2° , 13.3±0.2°, 17.2±0.2°, 23.8±0.2°, 25.5±0.2°, 27.1±0.2°, 15.3±0.2°, 15.8±0.2° or 21.4±0.2° at one or more diffraction peaks ,
    优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰。Preferably, there are diffraction peaks at optional 4, 5, 6, 8 or 10 positions.
  8. 根据权利要求4-7中任一项所述化合物的晶型，其特征在于，The crystal form of the compound according to any one of claims 4-7, characterized in that,
    晶型A具有如图1所示的X-射线粉末衍射图谱，或者具有如图2所示的DSC图谱；Form A has an X-ray powder diffraction pattern as shown in Figure 1, or has a DSC pattern as shown in Figure 2;
    晶型B具有如图3所示的X-射线粉末衍射图谱，或者具有如图4所示的DSC图谱；Form B has an X-ray powder diffraction pattern as shown in Figure 3, or has a DSC pattern as shown in Figure 4;
    晶型C具有如图5所示的X-射线粉末衍射图谱，或者具有如图6所示的DSC图谱；Form C has an X-ray powder diffraction spectrum as shown in Figure 5, or has a DSC spectrum as shown in Figure 6;
    晶型D具有如图7所示的X-射线粉末衍射图谱，或者具有如图8所示的DSC图谱。Form D has an X-ray powder diffraction pattern as shown in FIG. 7 , or a DSC pattern as shown in FIG. 8 .
  9. 根据权利要求1-3任一项所述化合物的晶型，其特征在于，所述晶型为N-(3'-(5-((6-乙酰基-2,6-二氮杂螺[3.3]庚烷-2-基)甲基)-6-甲氧基吡啶-2-基)-2,2'-二氯-[1,1'-联苯基]-3-基)-1,5-二甲基-4,5,6,7-四氢-1H-咪唑并[4,5-c]吡啶-2-甲酰胺的晶型B；The crystal form of the compound according to any one of claims 1-3, wherein the crystal form is N-(3'-(5-((6-acetyl-2,6-diazaspiro[ 3.3] Heptane-2-yl)methyl)-6-methoxypyridin-2-yl)-2,2'-dichloro-[1,1'-biphenyl]-3-yl)-1 , Form B of 5-dimethyl-4,5,6,7-tetrahydro-1H-imidazo[4,5-c]pyridine-2-carboxamide;
    所述晶型B的X-射线粉末衍射图谱在4.0±0.2°处具有衍射峰；或者在12.0±0.2°处具有衍射峰；或者在12.4±0.2°处具有衍射峰；或者在16.4±0.2°处具有衍射峰；或者在16.6±0.2°处具有衍射峰；或者在18.2±0.2°处具有衍射峰；或者在18.7±0.2°处具有衍射峰；或者在20.9±0.2°处具有衍射峰；或者在22.0±0.2°处具有衍射峰；或者在22.3±0.2°处具有衍射峰；优选包含上述衍射峰中的任意2-5处，或者3-5处，或者3-6处，或者3-8处，或者5-8处，或者6-8处，更优选包含其中任意6处、7处或8处；The X-ray powder diffraction pattern of the crystal form B has a diffraction peak at 4.0±0.2°; or has a diffraction peak at 12.0±0.2°; or has a diffraction peak at 12.4±0.2°; or has a diffraction peak at 16.4±0.2° or have a diffraction peak at 16.6±0.2°; or have a diffraction peak at 18.2±0.2°; or have a diffraction peak at 18.7±0.2°; or have a diffraction peak at 20.9±0.2°; or Have a diffraction peak at 22.0±0.2°; or have a diffraction peak at 22.3±0.2°; preferably include any 2-5, or 3-5, or 3-6, or 3-8 of the above-mentioned diffraction peaks , or 5-8, or 6-8, more preferably including any 6, 7 or 8 of them;
    优选地，所述晶型B的X-射线粉末衍射图谱至少包含位于2θ为4.0±0.2°、12.0±0.2°、12.4±0.2°中的一处或多处衍射峰，优选包含其中两条，更优选包 含三条；任选的，进一步还可以包含16.4±0.2°、16.6±0.2°、18.2±0.2°、18.7±0.2°、20.9±0.2°中的至少一条，优选包含其中2条、3条、4条或5条；Preferably, the X-ray powder diffraction pattern of the crystal form B includes at least one or more diffraction peaks at 2θ of 4.0±0.2°, 12.0±0.2°, and 12.4±0.2°, preferably two of them, More preferably, it contains three; optionally, it can further include at least one of 16.4±0.2°, 16.6±0.2°, 18.2±0.2°, 18.7±0.2°, 20.9±0.2°, preferably 2 or 3 of them , 4 or 5;
    更优选地，所述晶型B的X-射线粉末衍射图谱任选还包含位于2θ为22.0±0.2°、22.3±0.2°、22.7±0.2°、23.5±0.2°、26.5±0.2°、13.6±0.2°或14.3±0.2°处中的一处或多处衍射峰；优选至少包含其中任意2-3处，或者4-5处，或者6-7处；进一步优选，包含其中任意2处、3处、4处、5处、6处、7处；More preferably, the X-ray powder diffraction pattern of the crystal form B optionally further comprises One or more diffraction peaks at 0.2° or 14.3±0.2°; preferably at least include any 2-3, or 4-5, or 6-7 of them; more preferably, include any 2 or 3 of them place, 4 places, 5 places, 6 places, 7 places;
    进一步优选地，所述晶型B的X-射线粉末衍射图谱任选包含位于2θ为4.0±0.2°、12.0±0.2°、12.4±0.2°、16.4±0.2°、16.6±0.2°、18.2±0.2°、18.7±0.2°、20.9±0.2°、22.0±0.2°、22.3±0.2°、22.7±0.2°、23.5±0.2°、26.5±0.2°、13.6±0.2°或14.3±0.2°处中的一处或多处衍射峰，优选的，包含其中任选的4处、5处、6处、8处或10处有衍射峰；Further preferably, the X-ray powder diffraction pattern of the crystal form B optionally includes °, 18.7±0.2°, 20.9±0.2°, 22.0±0.2°, 22.3±0.2°, 22.7±0.2°, 23.5±0.2°, 26.5±0.2°, 13.6±0.2° or 14.3±0.2° One or more diffraction peaks, preferably, including optional 4, 5, 6, 8 or 10 diffraction peaks;
    更进一步优选地，所述晶型B的X-射线粉末衍射图谱如图10所示。Even more preferably, the X-ray powder diffraction pattern of the crystal form B is shown in FIG. 10 .
  10. 根据权利要求4-9任一项所述化合物的晶型，其特征在于，The crystal form of the compound according to any one of claims 4-9, characterized in that,
    晶型A、晶型B、晶型C和晶型D的X-射线粉末衍射图谱中相对峰强度为前十强的衍射峰位置与图1、图3、图5和图7对应位置衍射峰的2θ误差为±0.2°～±0.5°；优选±0.2°～±0.3°；最优选±0.2°。In the X-ray powder diffraction pattern of crystal form A, crystal form B, crystal form C and crystal form D, the relative peak intensity is the top ten diffraction peak positions and the corresponding positions of the diffraction peaks in Figure 1, Figure 3, Figure 5 and Figure 7 The 2θ error is ±0.2°～±0.5°; preferably ±0.2°～±0.3°; most preferably ±0.2°.
  11. 根据权利要求1-10任意一项所述化合物的晶型，其特征在于，晶型为无水物。The crystal form of the compound according to any one of claims 1-10, characterized in that the crystal form is an anhydrate.
  12. 根据权利要求1-11任意一项述化合物的晶型，其特征在于，晶型为水合物，水的个数为0.2-3；优选0.2、0.5、1、1.5、2、2.5或3；更优选0.5、1、2或3。According to the crystal form of the compound described in any one of claims 1-11, it is characterized in that the crystal form is a hydrate, and the number of water is 0.2-3; preferably 0.2, 0.5, 1, 1.5, 2, 2.5 or 3; more 0.5, 1, 2 or 3 are preferred.
  13. 一种制备权利要求1-12任意一项所述化合物晶型的方法，具体包括如下步骤：A method for preparing the crystal form of the compound described in any one of claims 1-12, specifically comprising the steps of:
    1)称取适量的自由碱，用良性溶剂在一定温度下溶解，温度优选0～50℃；1) Weigh an appropriate amount of free base and dissolve it in a benign solvent at a certain temperature, preferably 0-50°C;
    2)任选地，向以上所得溶液中加入反溶剂，搅拌至固体析出；2) Optionally, add an anti-solvent to the solution obtained above, and stir until solids are precipitated;
    3)任选地，搅拌，降温析晶得到目标产物；3) Optionally, stirring, cooling and crystallization to obtain the target product;
    或or
    1)称取适量的自由碱或其晶型，用不良溶剂在一定温度下打浆，温度优选0～50℃；1) Weighing an appropriate amount of free base or its crystal form, beating with a poor solvent at a certain temperature, preferably 0-50°C;
    其中：in:
    所述的良性溶剂选自甲醇、丙酮、甲酸乙酯、乙酸乙酯、乙腈、乙醇、88％ 丙酮、四氢呋喃、二氯甲烷、1,4-二氧六环、苯、甲苯、异丙醇、正丁醇、异丁醇、N，N-二甲基甲酰胺、N，N-二甲基乙酰胺、正丙醇、叔丁醇、2-丁酮或3-戊酮中的一种或多种；优选甲酸乙酯、异丙醇或无水乙醇中的一种或多种；Described benign solvent is selected from methanol, acetone, ethyl formate, ethyl acetate, acetonitrile, ethanol, 88% acetone, tetrahydrofuran, dichloromethane, 1,4-dioxane, benzene, toluene, Virahol, One of n-butanol, isobutanol, N,N-dimethylformamide, N,N-dimethylacetamide, n-propanol, tert-butanol, 2-butanone or 3-pentanone or Multiple; preferably one or more of ethyl formate, isopropanol or absolute ethanol;
    所述的反溶剂选自庚烷、环己烷、正己烷、正戊烷、水、乙酸乙酯、甲基叔丁基醚、甲苯或异丙醚中的一种或多种；优选乙酸乙酯、甲基叔丁基醚或环己烷中的一种或多种；The anti-solvent is selected from one or more of heptane, cyclohexane, n-hexane, n-pentane, water, ethyl acetate, methyl tert-butyl ether, toluene or isopropyl ether; preferred ethyl acetate One or more of esters, methyl tert-butyl ether or cyclohexane;
    所述的不良溶剂选自庚烷、环己烷、正己烷、正戊烷、水、乙酸乙酯、甲基叔丁基醚、甲苯或异丙醚中的一种或多种；优选乙酸乙酯、甲基叔丁基醚或环己烷中的一种或多种。Described poor solvent is selected from one or more in heptane, cyclohexane, normal hexane, normal pentane, water, ethyl acetate, methyl tert-butyl ether, toluene or isopropyl ether; Preferred ethyl acetate One or more of esters, methyl tert-butyl ether or cyclohexane.
  14. 一种药物组合物，其含有治疗有效量的权利要求1-12任一项所述化合物的晶型，以及一种或多种药学上可接受的载体、稀释剂或赋形剂。A pharmaceutical composition, which contains a therapeutically effective amount of the crystal form of the compound according to any one of claims 1-12, and one or more pharmaceutically acceptable carriers, diluents or excipients.
  15. 权利要求1-12任一项所述化合物的晶型或权利要求14所述的药物组合物在制备PD-1/PD-L1抑制剂药物中的应用。The application of the crystal form of the compound described in any one of claims 1-12 or the pharmaceutical composition described in claim 14 in the preparation of PD-1/PD-L1 inhibitor drugs.
  16. 权利要求1-12任一项所述化合物的晶型或权利要求14所述的药物组合物在制备治疗疾病选自癌症、感染性疾病、自身免疫性疾病药物中的应用；其中所述的癌症选自皮肤癌、肺癌、泌尿系肿瘤、血液肿瘤、乳腺癌、胶质瘤、消化系统肿瘤、生殖系统肿瘤、淋巴瘤、神经系统肿瘤、脑瘤、头颈癌；所述的感染性疾病选自细菌感染或病毒感染；所述的自身免疫性疾病选自器官特异性自身免疫病或系统性自身免疫病，其中，所述的器官特异性自身免疫病包括慢性淋巴细胞性甲状腺炎、甲状腺功能亢进、胰岛素依赖型糖尿病、重症肌无力、溃疡性结肠炎、恶性贫血伴慢性萎缩性胃炎、肺出血肾炎综合征、原发性胆汁性肝硬化、多发性脑脊髓硬化症或急性特发性多神经炎，所述的系统性自身免疫病包括类风湿关节炎、系统性红斑狼疮、系统性血管炎、硬皮病、天疱疮、皮肌炎、混合性结缔组织病或自身免疫性溶血性贫血。The application of the crystal form of the compound described in any one of claims 1-12 or the pharmaceutical composition described in claim 14 in the preparation of medicines for treating diseases selected from cancer, infectious diseases, and autoimmune diseases; wherein the cancer selected from skin cancer, lung cancer, urological tumor, blood tumor, breast cancer, glioma, digestive system tumor, reproductive system tumor, lymphoma, nervous system tumor, brain tumor, head and neck cancer; the infectious disease is selected from Bacterial infection or virus infection; Described autoimmune disease is selected from organ-specific autoimmune disease or systemic autoimmune disease, wherein, described organ-specific autoimmune disease comprises chronic lymphocytic thyroiditis, hyperthyroidism , insulin-dependent diabetes mellitus, myasthenia gravis, ulcerative colitis, pernicious anemia with chronic atrophic gastritis, pulmonary hemorrhage nephritic syndrome, primary biliary cirrhosis, multiple sclerosis or acute idiopathic polyneuropathy systemic autoimmune disease including rheumatoid arthritis, systemic lupus erythematosus, systemic vasculitis, scleroderma, pemphigus, dermatomyositis, mixed connective tissue disease, or autoimmune hemolytic anemia .
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Publication number Priority date Publication date Assignee Title
CN110267953A (en) * 2016-12-22 2019-09-20 因赛特公司 Imidazolidine simultaneously [4,5-C] pyridine derivate as PD-L1 internalization inducer
WO2020024997A1 (en) * 2018-08-01 2020-02-06 上海轶诺药业有限公司 Preparation and application of class of n-containing heterocyclic compounds having immunoregulatory function
WO2021136354A1 (en) * 2020-01-03 2021-07-08 上海翰森生物医药科技有限公司 Biphenyl derivative inhibitor, preparation method therefor and use thereof

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110267953A (en) * 2016-12-22 2019-09-20 因赛特公司 Imidazolidine simultaneously [4,5-C] pyridine derivate as PD-L1 internalization inducer
WO2020024997A1 (en) * 2018-08-01 2020-02-06 上海轶诺药业有限公司 Preparation and application of class of n-containing heterocyclic compounds having immunoregulatory function
WO2021136354A1 (en) * 2020-01-03 2021-07-08 上海翰森生物医药科技有限公司 Biphenyl derivative inhibitor, preparation method therefor and use thereof

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